Comparative proteomic analysis of hemolymph from uninfected and Candidatus Liberibacter asiaticus-infected Diaphorina citri.

Science.gov (United States)

Gill, T A; Chu, C; Pelz-Stelinski, K S

2017-02-01

Hemolymph was characterized from Diaphorina citri adults infected with the phytopathogen, Candidatus Liberibacter asiaticus (CLas), and compared with that from uninfected psyllids. This study identified 5531 and 3220 peptides within infected and uninfected hemolymph using nano-LC-MS/MS. A reduced number of proteins were detected for D. citri and all known endosymbionts within infected hemolymph as compared to uninfected hemolymph. A large number of immune defense proteins were absent from D. citri hemolymph; however, a single recognition protein (PGRP), two serine protease inhibitors, three prophenoloxidase (proPO) enzymes, and a single serine protease in an uninfected D. citri were detected. The hemolymph is nearly devoid of nutrient storage proteins. This is the first proteomic analysis of D. citri hemolymph that also analyses the components contributed by all the endosymbionts. By comparing the contribution of each endosymbiont (CCR, CPA, and WB) in the presence and absence of CLas infection, this study offers initial insights regarding the hemolymph response to microbial community shifts associated with D. citri infection status. Our data also present potential protein targets for analysis and disruption of CLas transmission that may facilitate management of huanglongbing (HLB) caused by CLas in citrus.

Secamiento Descendente del Yarumo (Cecropia Tessmanii

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Obregón Botero. Rafael

1972-12-01

Full Text Available El estudio de muchas enfermedades que afectan plantas tropicales, como en el caso del género Cecropia, es muy difícil, debido a la carencia de bibliografía, ya que en muchos géneros no se han adelantado estudios, ni siquiera someros, de sus parásitos o de sus dolencias de origen no parasitario. En el caso presente, no fue posible encontrar dato alguno que pudiera encaminarnos en el estudio de esta nueva enfermedad del yarumo. Creemos, a pesar de que el presente trabajo no representa un estudio completo de problema, que tiene dos aspectos importantes: 1.El iniciar el estudio de los enemigos que pueda tener una planta tan poco estudiada y a la vez de tantas posibilidades como fuente de materia prima. 2.Relacionar los parásitos de este género, con enfermedades de otras plantas que viven en esos mismos climas y medios ambientes, esas ya sí, de reconocida importancia económica, como son las del cacao, el caucho y algunos árboles maderables.

Isolation and partial purification of antimicrobial peptides/proteins from dung beetle, Onthophagus taurus immune hemolymph

International Nuclear Information System (INIS)

Vasanth Patil, H.B.; Sathish Kumar, B.Y.

2012-01-01

Antimicrobial peptides are important in the first line of the host defense system of all insect species. In the present study antimicrobial peptide(s) were isolated from the hemolymph of the dung beetle Onthophagus taurus. Both non induced and immune induced hemolymphs were tested for their antimicrobial activity against different bacterial strains and C. albicans. Induction was done by injecting E. coli into the abdominal cavity of the O. taurus. The non induced hemolymph did not show activity against any of the tested fungal and bacterial strains where as induced hemolymph showed activity against all tested bacterial strains but no activity against C. albicans. The induced hemolymph was subjected to non reducing SDS-PAGE and UV wavelength scan was performed to detect the presence of peptides. The immune induced hemolymph was purified by gel filtration chromatography to separate the proteins responsible for the antibacterial activity. The fractions within the peak were tested against those bacteria which previously showed sensitivity to the crude immune induced hemolymph. All fractions were found to be active against all tested bacteria with difference in zone of inhibition. The peptides are active against prokaryotes and not against eukaryotes. These properties reveal its unique characteristics and therapeutic application. (author)

Comparative proteomics analysis of silkworm hemolymph during the stages of metamorphosis via liquid chromatography and mass spectrometry.

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Hou, Yong; Zhang, Yan; Gong, Jing; Tian, Sha; Li, Jianwei; Dong, Zhaoming; Guo, Chao; Peng, Li; Zhao, Ping; Xia, Qingyou

2016-05-01

The silkworm is a lepidopteran insect that has an open circulatory system with hemolymph consisting of blood and lymph fluid. Hemolymph is not only considered as a depository of nutrients and energy, but it also plays a key role in substance transportation, immunity response, and proteolysis. In this study, we used LC-MS/MS to analyze the hemolymph proteins of four developmental stages during metamorphosis. A total of 728 proteins were identified from the hemolymph of the second day of wandering stage, first day of pupation, ninth day of pupation, and first day as an adult moth. GO annotations and categories showed that silkworm hemolymph proteins were enriched in carbohydrate metabolism, proteolysis, protein binding, and antibacterial humoral response. The levels of nutrient, immunity-related, and structural proteins changed significantly during development and metamorphosis. Some, such as cuticle, odorant-binding, and chemosensory proteins, showed stage-specific expression in the hemolymph. In addition, the expression of several antimicrobial peptides exhibited their highest level of abundance in the hemolymph of the early pupal stage. These findings provide a comprehensive proteomic insight of the silkworm hemolymph and suggest additional molecular targets for studying insect metamorphosis. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Hemolymph metabolites and osmolality are tightly linked to cold tolerance of Drosophila species

DEFF Research Database (Denmark)

Olsson, Trine; MacMillan, Heath A.; Nyberg, Nils

2016-01-01

osmolality was similar among all species despite chill-tolerant species having lower hemolymph [Na(+)]. Using NMR spectroscopy, we found that chill-tolerant species instead have higher levels of sugars and free amino acids in their hemolymph, including classical 'cryoprotectants' such as trehalose...

Free amino acids in spider hemolymph.

Science.gov (United States)

Tillinghast, Edward K; Townley, Mark A

2008-11-01

We examined the free amino acid composition of hemolymph from representatives of five spider families with an interest in knowing if the amino acid profile in the hemolymph of orb-web-building spiders reflects the high demands for small organic compounds in the sticky droplets of their webs. In nearly all analyses, on both orb and non-orb builders, glutamine was the most abundant free amino acid. Glycine, taurine, proline, histidine, and alanine also tended to be well-represented in orb and non-orb builders. While indications of taxon-specific differences in amino acid composition were observed, it was not apparent that two presumptive precursors (glutamine, taurine) of orb web sticky droplet compounds were uniquely enriched in araneids (orb builders). However, total amino acid concentrations were invariably highest in the araneids and especially so in overwintering juveniles, even as several of the essential amino acids declined during this winter diapause. Comparing the data from this study with those from earlier studies revealed a number of discrepancies. The possible origins of these differences are discussed.

Antibacterial activity in the hemolymph of the catarina scallop Argopecten ventricosus

OpenAIRE

Luna-González, Antonio; Maeda-Martínez, Alfonso; Campa-Córdova, Ángel; Orduña-Rojas, Javier

2007-01-01

We conducted a search for antibacterial peptide like activity in hemolymph of Argopecten ventricosus. Pre-purification of peptides was done by reverse phase HPLC. Hemolymph acidic supernatant was loaded into a column packed with C18 matrix. Stepwise elutions were performed with 5, 50, 80, and 100 % acetonitrile (ACN) in 0.05 % trifluoroacetic acid (TFA) over 550 min at a flow rate of 0.2 ml/min. Absorbance was monitored at 280 nm. Eluted fractions were concentrated under vacuum. Vibrio algino...

Antimicrobial lipids from the hemolymph of brachyuran crabs

Digital Repository Service at National Institute of Oceanography (India)

Ravichandran, S.; Wahidullah, S.; DeSouza, L.; Rameshkumar, G.

The potential of marine crabs as a source of biologically active products is largely unexplored. In the present study, antimicrobial activity of the hemolymph (plasma) and hemocytes (plasma cells) of six brachyuran crabs was investigated against 16...

Avaliação do potencial de utilização da madeira de Schizolobium amazonicum "Paricá" e Cecropia hololeuca "Embaúba" para produção de painéis aglomerados Evaluation of potential use of the wood of Schizolobium amazonicum "Paricá" and Cecropia hololeuca "Embaúba" to particleboard manufacture

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Setsuo Iwakiri

2010-01-01

Full Text Available Esta pesquisa teve por objetivo avaliar o potencial de utilização de madeira de Schizolobium amazonicum (Paricá e Cecropia hololeuca (Embaúba para produção de painéis aglomerados. Foram produzidos painéis experimentais com densidade nominal de 0,70 g/cm³, utilizando a resina uréia-formaldeído e partículas de madeira de Paricá e Embaúba, e mistura destas, em proporções de 75, 50 e 25%. A madeira de Pinus taeda foi utilizada como testemunha. Os painéis foram prensados com pressão específica de 40 kgf/cm², temperatura de 160ºC e tempo de prensagem de 8 minutos. Os resultados das avaliações de propriedades de absorção de água, inchamento em espessura, ligação interna, módulo de elasticidade e módulo de ruptura, indicaram que as madeiras de Schizolobium amazonicum (Paricá e Cecropia hololeuca (Embaúba são tecnicamente viáveis para produção de painéis aglomerados.This research was developed to evaluate the potential use of the woods Schizolobium amazonicum (Paricá and Cecropia hololeuca (Embaúba for manufacturing particleboard. Were made panels with the densities of 0.70 g/cm³, using mixtures of urea-formaldehyde resin and wood particles of Paricá and Embaúba, in proportions of 75, 50 and 25%. The Pinus taeda wood was used as the reference. The panels were pressed at the temperature of 160ºC, pressure of 40 kgf/cm², for 8 minutes. The evaluations of the properties of water absorption, thickness swelling, internal bond, modulus of elasticity and modulus of ruptures, showed that the woods of Schizolobium amazonicum (Paricá e Cecropia hololeuca (Embaúba are technically feasible for particleboard manufacture.

Cecropia peltata l.: Estudios farmacognósticos y de la composición de ácidos grasos libres

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Abel A Pardo Concepción

2000-08-01

Full Text Available Se inició el estudio de la yagruma (Cecropia peltata L. de uso medicinal en Cuba, con la descripción de los índices farmacognósticos mínimos necesarios para establecer la calidad de las hojas de la planta como droga, así como el estudio de la fracción de desengrase de donde se cristaliza una mezcla de 11 ácidos grasos metilados en forma libre, los cuales se caracterizan mediante cromatografía gaseosa acoplada a masas. El 50 % de estos ácidos son insaturados, lo cual puede favorecer el fundamento del uso de la planta popularmente con fines antiasmáticos.The study of yagruma (Cecropia peltata L. of medicinal use in Cuba was started with the description of the minimum pharmacognostic indexes necessary to establish the quality of the leaves of the plant as a drug, as well as the study of the scouring fraction from where a mix of 11 freely methylated fatty acids is crystallized. These acids are characterized by gas chromatography matched to masses. 50% of these acids are unsaturated, which may favor the popular use of the plant with antiasthmatic ends.

Culture-independent analysis of bacterial communities in hemolymph of American lobsters with epizootic shell disease.

Science.gov (United States)

Quinn, Robert A; Smolowitz, Roxanna; Chistoserdov, Andrei Y

2013-03-26

Epizootic shell disease (ESD) of the American lobster Homarus americanus H. Milne Edwards, 1837 is a disease of the carapace that presents grossly as large, melanized, irregularly shaped lesions, making the lobsters virtually unmarketable because of their grotesque appearance. We analyzed the bacterial communities present in the hemolymph of lobsters with and without ESD using nested-PCR of the 16S rRNA genes followed by denaturing gradient gel electrophoresis. All lobsters tested (n = 42) had bacterial communities in their hemolymph, and the community profiles were highly similar regardless of the sampling location or disease state. A number of bacteria were detected in a high proportion of samples and from numerous locations, including a Sediminibacterium sp. closely related to a symbiont of Tetraponera ants (38/42) and a Ralstonia sp. (27/42). Other bacteria commonly encountered included various Bacteroidetes, Pelomonas aquatica, and a Novosphingobium sp. One bacterium, a different Sediminibacterium sp., was detected in 20% of diseased animals (n = 29), but not in the lobsters without signs of ESD (n = 13). The bacteria in hemolymph were not the same as those known to be present in lesion communities except for the detection of a Thalassobius sp. in 1 individual. This work demonstrates that hemolymph bacteremia and the particular bacterial species present do not correlate with the incidence of ESD, providing further evidence that microbiologically, ESD is a strictly cuticular disease. Furthermore, the high incidence of the same species of bacteria in hemolymph of lobsters may indicate that they have a positive role in lobster fitness, rather than in disease, and further investigation of the role of bacteria in lobster hemolymph is required.

Enemigos naturales de Stenoma cecropia (Lepidoptera: Elachistidae) en palma de aceite, en el suroccidente de Colombia

OpenAIRE

SENDOYA-CORRALES, CARLOS ANDRÉS; BUSTILLO-PARDEY, ALEX ENRIQUE

2016-01-01

Stenoma cecropia infesta las nuevas siembras del híbrido interespecífico (Elaeis oleifera x E. guineensis) de palma de aceite en la zona de Tumaco (Nariño). Sus infestaciones afectan grandes extensiones de palma y causan defoliaciones de importancia económica. Por tal motivo, el objetivo de este trabajo fue determinar los factores bióticos y abióticos que afectan las poblaciones de la plaga a través de muestreos secuenciales quincenales, siguiendo un sistema móvil de desplazamiento 2 x 2, que...

Metabolism of labelled proteins of bombicid moth hemolymph at the final stage of its larval development

Energy Technology Data Exchange (ETDEWEB)

Klunova, S M; Altsybeeva, T I; Filippovich, Yu B [Moskovskij Gosudarstvennyj Pedagogicheskij Inst. (USSR)

1980-01-01

Studied was the distribution of radioactivity among hemolymph total proteins, fat body, carcass, intestinal wall, febroin and sericin sections of the silk gland after a single injection of hemolymph radioactive preparation into a bombyx. The fat body was the place of the synthesis of proteins used for silk protein formation at the end of 5-larval age.

Evaluation of potentical use of the wood of Schizolobium amazonicum “Paricá” and Cecropia hololeuca “Embaúba” for wood cement board manufacture

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Vinicius Yurk da Rocha

2012-06-01

Full Text Available This research was developed to evaluate the potential use of Schizolobium amazonicum “Paricá” and Cecropia hololeuca “Embaúba” wood for wood cement board manufacture. Panels with the density of 1.200 kg/m³ were made, using cement CP V ARI as mineral bonding and wood particles without treatment and treated with immersion in cold water, hot water and sodium hydroxide. The panels were pressed at the pressure of 40 kgf/cm², room temperature and press / clipping time of 24 hours and maturation time of 28 days. The evaluation of the properties of water absorption, thickness swelling, internal bond, modulus of elasticity and modulus of rupture, showed that Schizolobium amazonicum “Paricá” and Cecropia hololeuca “Embaúba” wood are technically feasible for wood-cement board manufacture. It was also found that the panels of embaúba exhibit some properties superior to those of the paricá, and the particles do not necessarily need treatment.

A serine protease inhibitor from hemolymph of green mussel, Perna viridis

Digital Repository Service at National Institute of Oceanography (India)

Khan, M.S.; Goswami, U.; Rojatkar, S.R.; Khan, M.I.

Bioactivity guided fractions of cell-free hemolymph of bacterially challenged marine mussel, Perna viridis led to the isolation of a novel quaternary alkaloid 1, which was identified by its spectral data. The isolated molecule 1 has been found...

Lab-on-a-chip and SDS-PAGE analysis of hemolymph protein profile from Rhipicephalus microplus (Acari: Ixodidae) infected with entomopathogenic nematode and fungus.

Science.gov (United States)

Golo, Patrícia Silva; Dos Santos, Alessa Siqueira de Oliveira; Monteiro, Caio Marcio Oliveira; Perinotto, Wendell Marcelo de Souza; Quinelato, Simone; Camargo, Mariana Guedes; de Sá, Fillipe Araujo; Angelo, Isabele da Costa; Martins, Marta Fonseca; Prata, Marcia Cristina de Azevedo; Bittencourt, Vânia Rita Elias Pinheiro

2016-09-01

In the present study, lab-on-a-chip electrophoresis (LoaC) was suggested as an alternative method to the conventional polyacrylamide gel electrophoresis under denaturing conditions (SDS-PAGE) to analyze raw cell-free tick hemolymph. Rhipicephalus microplus females were exposed to the entomopathogenic fungus Metarhizium anisopliae senso latu IBCB 116 strain and/or to the entomopathogenic nematode Heterorhabditis indica LPP1 strain. Hemolymph from not exposed or exposed ticks was collected 16 and 24 h after exposure and analyze by SDS-PAGE or LoaC. SDS-PAGE yielded 15 bands and LoaC electrophoresis 17 bands. Despite the differences in the number of bands, when the hemolymph protein profiles of exposed or unexposed ticks were compared in the same method, no suppressing or additional bands were detected among the treatments regardless the method (i.e., SDS-PAGE or chip electrophoresis using the Protein 230 Kit®). The potential of LoaC electrophoresis to detect protein bands from tick hemolymph was considered more efficient in comparison to the detection obtained using the traditional SDS-PAGE method, especially when it comes to protein subunits heavier than 100 KDa. LoaC electrophoresis provided a very good reproducibility, and is much faster than the conventional SDS-PAGE method, which requires several hours for one analysis. Despite both methods can be used to analyze tick hemolymph composition, LoaC was considered more suitable for cell-free hemolymph protein separation and detection. LoaC hemolymph band percent data reported changes in key proteins (i.e., HeLp and vitellogenin) exceptionally important for tick embryogenesis. This study reported, for the first time, tick hemolymph protein profile using LoaC.

Analyses of hemolymph from Amblyomma cajennense (Acari: ixodidae) using neutron activation analysis (NAA)

Energy Technology Data Exchange (ETDEWEB)

Simons, Simone M.; Oliveira, Daniella G.L.; Chudzinski-Tavassi, Ana M., E-mail: daniellaoliveira@butantan.gov.b, E-mail: amchudzinki@butantan.gov.b [Instituto Butantan, Sao Paulo, SP (Brazil); Zamboni, Cibele B., E-mail: czamboni@ipen.b [Instituto de Pesquisas Energeticas e Nucleares (IPEN/CNEN-SP), Sao Paulo, SP (Brazil)

2011-07-01

Instrumental neutron activation analysis technique (INAA) was applied to determine the elemental composition of hemolymph from Amblyomma cajennense tick. This biological material came from Butantan Institute (Sao Paulo city, Brazil) and it was investigated using the IEA-R1 nuclear reactor (4MW, pool type) at IPEN/CNEN-SP - Brazil. The concentration values for: Br (0.0032 {+-} 0.0005gL{sup -1}), Ca (0.104 {+-} 0.029gL{sup -1}), Cl (4.41 {+-} 0.25gL{sup -1}), I (76 {+-} 27{mu}gL{sup -1}), K (0.38 {+-} 0.09gL{sup -1}), Mg (0.038 {+-} 0.011gL{sup -1}), Na (4.30 {+-} 0.26gL{sup -1}) and S (1.35 {+-} 0.37gL{sup -1}) were determined for the first time. These data were compared with the concentration values established for Americanum and Anatolicum Excavatum tick species to clarify the ion balance in this biological material (hemolymph). This comparison suggests that Na concentration, majority in these species, has a similar behavior. These data also contribute to the understanding of hemolymph composition complementing its characterization as well as for the understanding of several physiological processes, especially those related to salivary secretion. (author)

Ovary histology and quantification of hemolymph proteins of Rhipicephalus (Boophilusmicroplus treated with Melia azedarach

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Lorena Alessandra Dias de Sousa

Full Text Available This study aimed to analyze ovary histology and quantify total protein in the hemolymph of Rhipicephalus (Boophilusmicroplus females treated with hexane extracts from green fruits of Melia azedarach. Eight engorged females were immersed in the extract at 0.25% concentration, and eight in water containing 5% acetone (control. The females were dissected 72 hours after treatment, and the ovaries were weighed and subjected to standard histological techniques. The total protein concentration was measured in the hemolymph of 200 females, of which 100 were treated as described above and 100 served as a control. In the treated group, ovary weight reduction and predominance of immature oocytes were observed. In addition, there were decreases in the diameters of the cytoplasm and germ vesicle of the oocytes in the treated group, compared with the controls. The protein concentration in the hemolymph was higher in the treated group than in the controls. The morphological changes observed in the treated ovaries included: presence of vacuolization; alteration of oocyte morphology, which changed from rounded to elongated; deformation of the chorion; and disorganization of the yolk granules. These results demonstrate the action ofM. azedarach fruit extracts on R.(B. microplus oogenesis.

[SPECTRAL AND ACID-BASE PROPERTIES OF HEMOLYMPH PLASMA AND ITS FRACTIONS FROM GASTROPOD PULMONATE MOLLUSC ACHATINA FULICA].

Science.gov (United States)

Petrova, T A; Lianguzov, A Yu; Malygina, N M

2016-01-01

The set of normal biochemical indicators of the hemolymph plasma of gastropod pulmonate mollusc Achatinafulica is described. Comparative analysis of the whole plasma and its subfractions enriched and depleted of oxygen-carrying protein hemocyanin was performed by spectrophotometry and spectrofluorimetry methods. Individual features of the absorption spectra were analyzed using fourth derivatives. The optimum method for estimating protein concentration was chosen. To characterize acid-base properties of plasma hemolymph and its sub-fractions we calculated buffer capacity, equivalence points and pK values of dominant buffer groups. It is shown that the major role in maintaining the buffer capacity of hemolymph belongs to the bicarbonate system. These results are compared with data for Helix pomatia available in literature. In the future the indicators studied in this work will be used to develop ecotoxicological criteria for the environmental assessment.

Fluconazole Pharmacokinetics in Galleria mellonella Larvae and Performance Evaluation of a Bioassay Compared to Liquid Chromatography-Tandem Mass Spectrometry for Hemolymph Specimens

Science.gov (United States)

Astvad, Karen Marie Thyssen; Meletiadis, Joseph; Whalley, Sarah

2017-01-01

ABSTRACT The invertebrate model organism Galleria mellonella can be used to assess the efficacy of treatment of fungal infection. The fluconazole dose best mimicking human exposure during licensed dosing is unknown. We validated a bioassay for fluconazole detection in hemolymph and determined the fluconazole pharmacokinetics and pharmacodynamics in larval hemolymph in order to estimate a humanized dose for future experiments. A bioassay using 4-mm agar wells, 20 μl hemolymph, and the hypersusceptible Candida albicans DSY2621 was established and compared to a validated liquid chromatography-tandem mass spectrometry (LC–MS-MS) method. G. mellonella larvae were injected with fluconazole (5, 10, and 20 mg/kg of larval weight), and hemolymph was harvested for 24 h for pharmacokinetics calculations. The exposure was compared to the human exposure during standard licensed dosing. The bioassay had a linear standard curve between 1 and 20 mg/liter. Accuracy and coefficients of variation (percent) values were below 10%. The Spearman coefficient between assays was 0.94. Fluconazole larval pharmacokinetics followed one-compartment linear kinetics, with the 24-h area under the hemolymph concentration-time curve (AUC24 h) being 93, 173, and 406 mg · h/liter for the three doses compared to 400 mg · h/liter in humans under licensed treatment. In conclusion, a bioassay was validated for fluconazole determination in hemolymph. The pharmacokinetics was linear. An exposure comparable to the human exposure during standard licensed dosing was obtained with 20 mg/kg. PMID:28760893

Fluconazole Pharmacokinetics in Galleria mellonella Larvae and Performance Evaluation of a Bioassay Compared to Liquid Chromatography-Tandem Mass Spectrometry for Hemolymph Specimens

DEFF Research Database (Denmark)

Astvad, Karen Marie Thyssen; Meletiadis, Joseph; Whalley, Sarah

2017-01-01

The invertebrate model organism Galleria mellonella can be used to assess the efficacy of treatment of fungal infection. The fluconazole dose best mimicking human exposure during licensed dosing is unknown. We validated a bioassay for fluconazole detection in hemolymph and determined...... the fluconazole pharmacokinetics and pharmacodynamics in larval hemolymph in order to estimate a humanized dose for future experiments. A bioassay using 4-mm agar wells, 20 μl hemolymph, and the hypersusceptible Candida albicans DSY2621 was established and compared to a validated liquid chromatography-tandem mass...... spectrometry (LC-MS-MS) method. G. mellonella larvae were injected with fluconazole (5, 10, and 20 mg/kg of larval weight), and hemolymph was harvested for 24 h for pharmacokinetics calculations. The exposure was compared to the human exposure during standard licensed dosing. The bioassay had a linear standard...

THE REFERENCE VALUES OF THE MAIN BIOCHEMICAL PARAMETERS OF THE HEMOLYMPH OF APIS MELLIFERA CARPATHICA IN SOUTH-EASTERN ROMANIA

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AGRIPINA SAPCALIU

2007-10-01

Full Text Available Although biochemical analyses of the blood were and are still used for the routinediagnosis and especially for the metabolic survey in farm animals, such analyses, maybe applied for the honeybee hemolymph (as a paraclinic examination. The aim of thisexperimental study was to investigate and to determine the reference values of themain biochemical parameters in the hemolymph of the healthy honeybees of Apismellifera carpathica. The honeybee samples were collected in order to analyze thehealth condition of the respective colony. All the samples coming from sick colonieswere removed, only healthy adult honeybees coming from strong colonies were keptand used for hemolymph collection. By special methods, samples of hemolymph (300μl/sample collected from about 50 individuals, were analyzed both during the activeseason (spring/summer and the inactive season (autumn/winter. The study wascarried out on 50 samples of undiluted hemolymph taken from a total number of about2,500 honeybees. The following 21 biochemical parameters were analyzed: GLU(mg/dl, HDL-c (mg/dl, ALP (UI/l, T-cho (mg/dl, Tprot (mg/dl, Alb. (g/dl, BUN(mg/dl, LDH (UI/l, CPK (UI/l, Mg (mg/dl, FRA (μm/l, IP (mg/dl, GGT (UI/l,GOT (UI/l, GPT (UI/l, Ca (mg/dl, Cre (mg/dl, Amy (UI/l, T–BIL (mg/dl, TG(mg/dl, UA (mg/dl. The test was carried out after the collection and processing of thesamples using the SPOTCHEM EZSP4430, equipment with dry kits, the slides technique,respectively.

Larval hemolymph of rhinoceros beetle, Allomyrina dichotoma, enhances insulin secretion through ATF3 gene expression in INS-1 pancreatic β-cells.

Science.gov (United States)

Kim, Seung-Whan; Suh, Hyun-Woo; Yoo, Bo-Kyung; Kwon, Kisang; Yu, Kweon; Choi, Ji-Young; Kwon, O-Yu

2018-05-22

In this study, we show that INS-1 pancreatic β-cells treated for 2 h with hemolymph of larvae of rhinoceros beetle, Allomyrina dichotoma, secreted about twice as much insulin compared to control cells without such treatment. Activating transcription factor 3 (ATF3) was the highest upregulated gene in DNA chip analysis. The A. dichotoma hemolymph dose-dependently induced increased expression levels of genes encoding ATF3 and insulin. Conversely, treatment with ATF3 siRNA inhibited expression levels of both genes and curbed insulin secretion. These results suggest that the A. dichotoma hemolymph has potential for treating and preventing diabetes or diabetes-related complications.

Herança e desequilíbrio de ligação em sete locos isoenzimáticos de Cecropia pachystachya. Inheritance and linkage disequilibrium in sevem isozymes loci of Cecropia pachystachya.

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Luciano Arruda RIBAS

2004-12-01

Full Text Available Seis sistemas enzimáticos (ACP,G6PDH, IDH, PGI, PGM e SKDH, codificandosete locos polimórficos (Acp-3, G6pdh-1, Idh-1,Pgi-2, Pgm-1, Pgm-2 e Skdh-1, foram investigadospara a herança e desequilíbrio de ligação emCecropia pachystachya Trec. A herança mendelianafoi confirmada para todos os locos avaliados.Desvios significativos da razão de segregaçãoesperada 1:1 foram detectados apenas em umaprogênies de árvore heterozigótica. O desequilíbriode ligação foi examinado para 21 pares de locosisoenzimáticos, não sendo detectados desequilíbriosentre nenhum deles e, assim, estes podem serutilizados em subseqüentes estudos do sistemade reprodução, diversidade e estrutura genética deC. pachystachya.

Effects of lead on Na+, K+-ATPase and hemolymph ion concentrations in the freshwater mussel Elliptio complanata

Science.gov (United States)

Mosher, Shad; Cope, W. Gregory; Weber, Frank X.; Shea, Damian; Kwak, Thomas J.

2012-01-01

Freshwater mussels are an imperiled fauna exposed to a variety of environmental toxicants such as lead (Pb) and studies are urgently needed to assess their health and condition to guide conservation efforts. A 28-day laboratory toxicity test with Pb and adult Eastern elliptio mussels (Elliptio complanata) was conducted to determine uptake kinetics and to assess the toxicological effects of Pb exposure. Test mussels were collected from a relatively uncontaminated reference site and exposed to a water-only control and five concentrations of Pb (as lead nitrate) ranging from 1 to 245 mu g/L in a static renewal test with a water hardness of 42 mg/L. Endpoints included tissue Pb concentrations, hemolymph Pb and ion (Na+, K+, Cl-, Ca2+) concentrations, and Na+, K+-ATPase enzyme activity in gill tissue. Mussels accumulated Pb rapidly, with tissue concentrations increasing at an exposure-dependent rate for the first 2 weeks, but with no significant increase from 2 to 4 weeks. Mussel tissue Pb concentrations ranged from 0.34 to 898 mu g/g dry weight, were strongly related to Pb in test water at every time interval (7, 14, 21, and 28 days), and did not significantly increase after day 14. Hemolymph Pb concentration was variable, dependent on exposure concentration, and showed no appreciable change with time beyond day 7, except for mussels in the greatest exposure concentration (245 mu g/L), which showed a significant reduction in Pb by 28 days, suggesting a threshold for Pb binding or elimination in hemolymph at concentrations near 1000 mu g/g. The Na+, K+-ATPase activity in the gill tissue of mussels was significantly reduced by Pb on day 28 and was highly correlated with tissue Pb concentration (R2 = 0.92; P = 0.013). The Na+, K+-ATPase activity was correlated with reduced hemolymph Na+ concentration at the greatest Pb exposure when enzyme activity was at 30% of controls. Hemolymph Ca2+ concentration increased significantly in mussels from the greatest Pb exposure and may

Hemolymph proteome changes during worker brood development match the biological divergences between western honey bees (Apis mellifera) and eastern honey bees (Apis cerana).

Science.gov (United States)

Feng, Mao; Ramadan, Haitham; Han, Bin; Fang, Yu; Li, Jianke

2014-07-05

Hemolymph plays key roles in honey bee molecule transport, immune defense, and in monitoring the physiological condition. There is a lack of knowledge regarding how the proteome achieves these biological missions for both the western and eastern honey bees (Apis mellifera and Apis cerana). A time-resolved proteome was compared using two-dimensional electrophoresis-based proteomics to reveal the mechanistic differences by analysis of hemolymph proteome changes between the worker bees of two bee species during the larval to pupal stages. The brood body weight of Apis mellifera was significantly heavier than that of Apis cerana at each developmental stage. Significantly, different protein expression patterns and metabolic pathways were observed in 74 proteins (166 spots) that were differentially abundant between the two bee species. The function of hemolymph in energy storage, odor communication, and antioxidation is of equal importance for the western and eastern bees, indicated by the enhanced expression of different protein species. However, stronger expression of protein folding, cytoskeletal and developmental proteins, and more highly activated energy producing pathways in western bees suggests that the different bee species have developed unique strategies to match their specific physiology using hemolymph to deliver nutrients and in immune defense. Our disparate findings constitute a proof-of-concept of molecular details that the ecologically shaped different physiological conditions of different bee species match with the hemolymph proteome during the brood stage. This also provides a starting point for future research on the specific hemolymph proteins or pathways related to the differential phenotypes or physiology.

An HPLC-DAD method to quantification of main phenolic compounds from leaves of Cecropia species

Energy Technology Data Exchange (ETDEWEB)

Costa, Geison M.; Ortmann, Caroline F.; Schenkel, Eloir P.; Reginatto, Flavio H., E-mail: freginatto@hotmail.co [Universidade Federal de Santa Catarina (UFSC), Florianopolis (Brazil). Centro de Ciencias da Saude. Dept. de Ciencias Farmaceuticas

2011-07-01

An efficient and reproducible HPLC-DAD method was developed and validated for the simultaneous quantification of major compounds (chlorogenic acid, isoorientin, orientin and isovitexin) present in the leaves of two Cecropia species, C. glaziovii and C. pachystachya. From the leaves of C. glaziovii and C. pachystachya were isolated the C-glycosylflavones isoorientin and isovitexin and identified on both species chlorogenic acid (3-O-caffeoylquinic acid) and the O-glycosylflavonol isoquercitrin. The C-glycosylflavone orientin was isolated only from C. pachystachya. Chlorogenic acid was the major compound in both species (11.1 mg g{sup -1} of extract of C. glaziovii and 27.2 mg g{sup -1} of extract of C. pachystachya) and for the flavonoids quantified, isovitexin was the main C-glycosylflavonoid for C. glaziovii (4.6 mg g{sup -1} of extract) and isoorientin the main one for C. pachystachya (17.3 mg g{sup -1} of extract). (author)

An HPLC-DAD method to quantification of main phenolic compounds from leaves of Cecropia species

International Nuclear Information System (INIS)

Costa, Geison M.; Ortmann, Caroline F.; Schenkel, Eloir P.; Reginatto, Flavio H.

2011-01-01

An efficient and reproducible HPLC-DAD method was developed and validated for the simultaneous quantification of major compounds (chlorogenic acid, isoorientin, orientin and isovitexin) present in the leaves of two Cecropia species, C. glaziovii and C. pachystachya. From the leaves of C. glaziovii and C. pachystachya were isolated the C-glycosylflavones isoorientin and isovitexin and identified on both species chlorogenic acid (3-O-caffeoylquinic acid) and the O-glycosylflavonol isoquercitrin. The C-glycosylflavone orientin was isolated only from C. pachystachya. Chlorogenic acid was the major compound in both species (11.1 mg g -1 of extract of C. glaziovii and 27.2 mg g -1 of extract of C. pachystachya) and for the flavonoids quantified, isovitexin was the main C-glycosylflavonoid for C. glaziovii (4.6 mg g -1 of extract) and isoorientin the main one for C. pachystachya (17.3 mg g -1 of extract). (author)

Effect of salinity on hemolymph osmotic pressure, sodium concentration and Na+-K+-ATPase activity of gill of Chinese crab, Eriocheir sinensis

Science.gov (United States)

Liu, Hongyu; Pan, Luqing; Fu, Lü

2008-02-01

The effects of salinity on hemolymph osmotic pressure, Na+ concentration and Na+-K+-ATPase activity of gill of Chinese crab Eriocheir sinensis were studied. The results showed that hemolymph osmotic pressure and Na+ concentration increased significantly ( P0.05); However, the protein concentration decreased gradually with the increase of salinity from 0.25 d to 1 d, and then tended to be stable from day 1 to day 15.

Evaluation of potential use of the wood of Schizolobium amazonicum "Paricá" and Cecropia hololeuca "Embaúba" to particleboard manufacture

OpenAIRE

Iwakiri, Setsuo; Zeller, Florian; Pinto, Juliana Afonso; Ramirez, Maria Guadalupe Lomeli; Souza, Marina Moura; Seixas, Rodrigo

2010-01-01

Esta pesquisa teve por objetivo avaliar o potencial de utilização de madeira de Schizolobium amazonicum (Paricá) e Cecropia hololeuca (Embaúba) para produção de painéis aglomerados. Foram produzidos painéis experimentais com densidade nominal de 0,70 g/cm³, utilizando a resina uréia-formaldeído e partículas de madeira de Paricá e Embaúba, e mistura destas, em proporções de 75, 50 e 25%. A madeira de Pinus taeda foi utilizada como testemunha. Os painéis foram prensados com pressão específica d...

Hemolymph chemistry and histopathological changes in Pacific oysters (Crassostrea gigas) in response to low salinity stress.

Science.gov (United States)

Knowles, Graeme; Handlinger, Judith; Jones, Brian; Moltschaniwskyj, Natalie

2014-09-01

This study described seasonal differences in the histopathological and hemolymph chemistry changes in different family lines of Pacific oysters, Crassostrea gigas, in response to the stress of an abrupt change to low salinity, and mechanical grading. The most significant changes in pallial cavity salinity, hemolymph chemistry and histopathological findings occurred in summer at low salinity. In summer (water temperature 18°C) at low salinity, 9 (25.7% of full salinity), the mean pallial cavity salinity in oysters at day 3 was 19.8±1.6 (SE) and day 10 was 22.8±1.6 (SE) lower than oysters at salinity 35. Associated with this fall in pallial cavity salinity, mean hemolymph sodium for oysters at salinity 9 on day 3 and 10 were 297.2mmol/L±20(SE) and 350.4mmol/L±21.3(SE) lower than oysters at salinity 35. Similarly mean hemolymph potassium in oysters held at salinity 9 at day 3 and 10 were 5.6mmol/L±0.6(SE) and 7.9mmol/L±0.6 (SE) lower than oysters at salinity 35. These oysters at low salinity had expanded intercellular spaces and significant intracytoplasmic vacuolation distending the cytoplasm of epithelial cells in the alimentary tract and kidney and hemocyte infiltrate (diapedesis) within the alimentary tract wall. In contrast, in winter (water temperature 8°C) oyster mean pallial cavity salinity only fell at day 10 and this was by 6.0±0.6 (SE) compared to that of oysters at salinity 35. There were limited histopathological changes (expanded intercellular spaces and moderate intracytoplasmic vacuolation of renal epithelial cells) in these oysters at day 10 in low salinity. Mechanical grading and family line did not influence the oyster response to sudden low salinity. These findings provide additional information for interpretation of non-lethal, histopathological changes associated with temperature and salinity variation. Copyright © 2014 Elsevier Inc. All rights reserved.

Development of chemical isotope labeling liquid chromatography mass spectrometry for silkworm hemolymph metabolomics

International Nuclear Information System (INIS)

Shen, Weifeng; Han, Wei; Li, Yunong; Meng, Zhiqi; Cai, Leiming; Li, Liang

2016-01-01

Silkworm (Bombyx mori) is a very useful target insect for evaluation of endocrine disruptor chemicals (EDCs) due to mature breeding techniques, complete endocrine system and broad basic knowledge on developmental biology. Comparative metabolomics of silkworms with and without EDC exposure offers another dimension of studying EDCs. In this work, we report a workflow on metabolomic profiling of silkworm hemolymph based on high-performance chemical isotope labeling (CIL) liquid chromatography mass spectrometry (LC-MS) and demonstrate its application in studying the metabolic changes associated with the pesticide dichlorodiphenyltrichloroethane (DDT) exposure in silkworm. Hemolymph samples were taken from mature silkworms after growing on diet that contained DDT at four different concentrations (1, 0.1, 0.01, 0.001 ppm) as well as on diet without DDT as controls. They were subjected to differential "1"2C-/"1"3C-dansyl labeling of the amine/phenol submetabolome, LC-UV quantification of the total amount of labeled metabolites for sample normalization, and LC-MS detection and relative quantification of individual metabolites in comparative samples. The total concentration of labeled metabolites did not show any significant change between four DDT-treatment groups and one control group. Multivariate statistical analysis of the metabolome data set showed that there was a distinct metabolomic separation between the five groups. Out of the 2044 detected peak pairs, 338 and 1471 metabolites have been putatively identified against the HMDB database and the EML library, respectively. 65 metabolites were identified by the dansyl library searching based on the accurate mass and retention time. Among the 65 identified metabolites, 33 positive metabolites had changes of greater than 1.20-fold or less than 0.83-fold in one or more groups with p-value of smaller than 0.05. Several useful biomarkers including serine, methionine, tryptophan, asymmetric dimethylarginine, N

Development of chemical isotope labeling liquid chromatography mass spectrometry for silkworm hemolymph metabolomics

Energy Technology Data Exchange (ETDEWEB)

Shen, Weifeng [Key Laboratory of Detection for Pesticide Residues, Ministry of Agriculture (China); Sericultural Research Institute, Zhejiang Academy of Agricultural Sciences, Hangzhou (China); Han, Wei; Li, Yunong [Department of Chemistry, University of Alberta, Edmonton, Alberta (Canada); Meng, Zhiqi [Sericultural Research Institute, Zhejiang Academy of Agricultural Sciences, Hangzhou (China); Cai, Leiming, E-mail: cailm@mail.zaas.ac.cn [Institute of Quality and Standard for Agro-products, Zhejiang Academy of Agricultural Sciences, Hangzhou (China); Li, Liang, E-mail: Liang.Li@ualberta.ca [Department of Chemistry, University of Alberta, Edmonton, Alberta (Canada)

2016-10-26

Silkworm (Bombyx mori) is a very useful target insect for evaluation of endocrine disruptor chemicals (EDCs) due to mature breeding techniques, complete endocrine system and broad basic knowledge on developmental biology. Comparative metabolomics of silkworms with and without EDC exposure offers another dimension of studying EDCs. In this work, we report a workflow on metabolomic profiling of silkworm hemolymph based on high-performance chemical isotope labeling (CIL) liquid chromatography mass spectrometry (LC-MS) and demonstrate its application in studying the metabolic changes associated with the pesticide dichlorodiphenyltrichloroethane (DDT) exposure in silkworm. Hemolymph samples were taken from mature silkworms after growing on diet that contained DDT at four different concentrations (1, 0.1, 0.01, 0.001 ppm) as well as on diet without DDT as controls. They were subjected to differential {sup 12}C-/{sup 13}C-dansyl labeling of the amine/phenol submetabolome, LC-UV quantification of the total amount of labeled metabolites for sample normalization, and LC-MS detection and relative quantification of individual metabolites in comparative samples. The total concentration of labeled metabolites did not show any significant change between four DDT-treatment groups and one control group. Multivariate statistical analysis of the metabolome data set showed that there was a distinct metabolomic separation between the five groups. Out of the 2044 detected peak pairs, 338 and 1471 metabolites have been putatively identified against the HMDB database and the EML library, respectively. 65 metabolites were identified by the dansyl library searching based on the accurate mass and retention time. Among the 65 identified metabolites, 33 positive metabolites had changes of greater than 1.20-fold or less than 0.83-fold in one or more groups with p-value of smaller than 0.05. Several useful biomarkers including serine, methionine, tryptophan, asymmetric dimethylarginine, N

Fat body, hemolymph and ovary routes for delivery of substances to ovary in Melipona quadrifasciata anthidioides: differences among castes through the use of electron-opaque tracers.

Science.gov (United States)

da Cruz-Landim, Carminda; Roat, Thaisa Cristina; Berger, Bruno

2013-08-01

The yolk protein precursor, vitellogenin (Vg), in bees is synthesized in the fat body trophocytes, delivered to the hemolymph and ultimately absorbed from there during the vitellogenic phase of oocytes in the active ovary. The routes tracing the material exchange that occurs between the trophocytes and the hemolymph, in addition to the transportation from the hemolymph to the ovarian follicles, were marked by alkaline phosphatase and lanthanum nitrate (LN). Active ovaries from nurse workers and physogastric queens, as well as inactive ovaries of virgin queens, were examined by transmission electron microscopy. The LN permitted better visualization of the routes of exchanges between the organs and the hemolymph. Both methods demonstrate the apparent differences between the phases of the ovary and the bee caste. In inactive ovaries of the virgin queens, the routes from the follicular epithelium to the oocyte remain closed; conversely, they are open in active ovaries of the nurse workers and physogastric queens. The differences between the methods and classes of bees are discussed.

Insect biofuel cells using trehalose included in insect hemolymph leading to an insect-mountable biofuel cell.

Science.gov (United States)

Shoji, Kan; Akiyama, Yoshitake; Suzuki, Masato; Hoshino, Takayuki; Nakamura, Nobuhumi; Ohno, Hiroyuki; Morishima, Keisuke

2012-12-01

In this paper, an insect biofuel cell (BFC) using trehalose included in insect hemolymph was developed. The insect BFC is based on trehalase and glucose oxidase (GOD) reaction systems which oxidize β-glucose obtained by hydrolyzing trehalose. First, we confirmed by LC-MS that a sufficient amount of trehalose was present in the cockroach hemolymph (CHL). The maximum power density obtained using the insect BFC was 6.07 μW/cm(2). The power output was kept more than 10 % for 2.5 h by protecting the electrodes with a dialysis membrane. Furthermore, the maximum power density was increased to 10.5 μW/cm(2) by using an air diffusion cathode. Finally, we succeeded in driving a melody integrated circuit (IC) and a piezo speaker by connecting five insect BFCs in series. The results indicate that the insect BFC is a promising insect-mountable battery to power environmental monitoring micro-tools.

Total cysteine and glutathione determination in hemolymph of individual adult D. melanogaster

Energy Technology Data Exchange (ETDEWEB)

Borra, Srivani, E-mail: sborra3@uic.edu [Department of Chemistry, University of Illinois at Chicago, 845 West Taylor Street, 4323 SES, MC 111, Chicago, IL 60607 (United States); Featherstone, David E., E-mail: def@uic.edu [Department of Biological Sciences, University of Illinois at Chicago, 840 West Taylor Street, SEL 4311, M/C 067, Chicago, IL 60607 (United States); Laboratory of Integrative Neuroscience, University of Illinois at Chicago, 840 West Taylor Street, SEL 4311, M/C 067, Chicago, IL 60607 (United States); Shippy, Scott A., E-mail: sshippy@uic.edu [Department of Chemistry, University of Illinois at Chicago, 845 West Taylor Street, 5417 SES, MC 111, Chicago, IL 60607 (United States); Laboratory of Integrative Neuroscience, University of Illinois at Chicago, 840 West Taylor Street, SEL 4311, M/C 067, Chicago, IL 60607 (United States)

2015-01-01

Highlights: • Method for highly volume variant, nL sample assay of biological relevant thiols. • Defined capillary lengths used to deliver nL sample and reagent volumes. • Optimized reagent concentrations, reaction times and temperatures for thiol assay. • Total cysteine and glutathione measured from hemolymph of individual fruit flies. - Abstract: Determination of thiols, glutathione (GSH) and cysteine (Cys) are important due to their roles in oxidative stress and aging. Oxidants such as soluble O{sub 2} and H{sub 2}O{sub 2} promote oxidation of thiols to disulfide (-S-S-) bonded dimers affecting quantitation accuracy. The method presented here reduces disulfide-bonded species followed by fluorescence labelling of the 29.5 (±18.2) nL hemolymph volumes of individual adult Drosophila Melanogaster. The availability of only tens of nanoliter (nL) samples that are also highly volume variant requires efficient sample handling to improve thiol measurements while minimizing sample dilution. The optimized method presented here utilizes defined lengths of capillaries to meter tris(2-carboxyethyl)phosphine reducing reagent and monobromobimane derivatizing reagent volumes enabling Cys and GSH quantitation with only 20-fold dilution. The nL assay developed here was optimized with respect to reagent concentrations, sample dilution, reaction times and temperatures. Separation and identification of the nL thiol mixtures were obtained with capillary electrophoresis-laser induced fluorescence. To demonstrate the capability of this method total Cys and total GSH were measured in the hemolymph collected from individual adult D. Melanogaster. The thiol measurements were used to compare a mutant fly strain with a non-functional cystine–glutamate transporter (xCT) to its background control. The mutant fly, genderblind (gb), carries a non-functional gene for a protein similar to mammalian xCT whose function is not fully understood. Average concentrations obtained for mutant

Some triterpenic compounds in extracts of Cecropia and Bauhinia species for different sampling years

Directory of Open Access Journals (Sweden)

Marcella Emilia Pietra Schmidt

Full Text Available ABSTRACT The aim of this paper is to provide an overview on the chemical composition of triterpenes in widespread used folk medicine species, through the development and validation of eleven compounds using HPLC-UV detection. The compounds were separated using isocratic elution, on a reverse phase column (Kinetex C18, 250 mm × 4.6 mm, 5 µm with mobile phase consisted of acetonitrile:tetrahydrofuran (90:10, v/v, flow-rate of 0.5 ml/min and detection in 210 nm. Diverse validation parameters were successfully evaluated. The samples of Bauhinia variegata L., B. variegata var. candida Voigt, Fabaceae, Cecropia palmata Willd. and C. obtusa Trécul, Urticaceae, collected in 2012, 2013 and 2014 from Amazon were treated with two different solvents (ethyl acetate and chloroform and analyzed by the proposed method. Stigmasterol, lupeol, β-sitosterol, β-amirin and α-amirin were found in all the studied plants. Highlighting the presence of oleanolic acid, maslinic acid in C. obtusa and C. palmata extracts, erythrodiol only in C. palmata, stigmasteol in B. variegata and α-amirin in B. variegata var. candida. Overall, ethyl acetate showed better performance as the extractor solvent than chloroform. Moreover, it could be used for the quality control of medicinal plants and to assess potential marker compounds.

"Phloem sap analysis of Schleichera oleosa (Lour) Oken, Butea monosperma (Lam) Taub. and Ziziphus mauritiana (Lam) and hemolymph of Kerria lacca (Kerr) using HPLC and tandem mass spectrometry".

Science.gov (United States)

Vashishtha, Amit; Rathi, Brijesh; Kaushik, Sandeep; Sharma, K K; Lakhanpaul, Suman

2013-10-01

Females of lac insects especially of Kerria lacca (Kerr) secret a resin known as lac for their own protection, which has tremendous applications. Lac insect completes its lifecycle on several host taxa where it exclusively feeds on phloem sap but Schleichera oleosa (Lour.) Oken, Butea monosperma (Lam.) and Ziziphus mauritiana (Lam.) are its major hosts. Analysis of phloem sap constituents as well as hemolymph of lac insect is important because it ultimately gets converted into lac by insect intervention. Main phloem sap constituent's viz. sugars and free amino acids and hemolymph of lac insect were analyzed using HPLC and tandem mass spectrometry, respectively. The results were transformed to relative percentage of the total sugars and free amino acids analyzed in each sample for comparison among lac insect hemolymph and the phloem sap of the three different host taxa. Sucrose (58.9 ± 3.6-85.6 ± 0.9) and trehalose (62.3 ± 0.4) were the predominant sugars in phloem sap of three taxa and hemolymph of lac insect, respectively. Glutamic acid (33.1 ± 1.4-39.8 ± 1.4) was found to be main amino acid among the phloem sap of three taxa while tyrosine (61 ± 2.6) was the major amino acid in hemolymph of lac insect. The relative percentage of non-essential amino acids (60.8 %-69.9 %) was found to be more in all the three host taxa while essential amino acids (30.1 %-35.4 %) were present at a lower relative percentage. In contrast to this, the relative percentage of essential amino acids (81.9 %) was observed to be higher as compared to non-essential amino acids (17.7 %) in lac insect hemolymph. These results led to the detection of lac insect's endosymbionts. Moreover, this study revealed a clue regarding the importance of development of a synthetic diet for this insect so that a precise pathway of lac biosynthesis could be investigated for thorough understanding.

A mosquito hemolymph odorant-binding protein family member specifically binds juvenile hormone

Energy Technology Data Exchange (ETDEWEB)

Kim, Il Hwan; Pham, Van; Jablonka, Willy; Goodman, Walter G.; Ribeiro, José M. C.; Andersen, John F.

2017-07-27

Juvenile hormone (JH) is a key regulator of insect development and reproduction. In adult mosquitoes, it is essential for maturation of the ovary and normal male reproductive behavior, but how JH distribution and activity is regulated after secretion is unclear. Here, we report a new type of specific JH-binding protein, given the name mosquito juvenile hormone-binding protein (mJHBP), which circulates in the hemolymph of pupal and adult Aedes aegypti males and females. mJHBP is a member of the odorant-binding protein (OBP) family, and orthologs are present in the genomes of Aedes, Culex, and Anopheles mosquito species. Using isothermal titration calorimetry, we show that mJHBP specifically binds JH II and JH III but not eicosanoids or JH derivatives. mJHBP was crystallized in the presence of JH III and found to have a double OBP domain structure reminiscent of salivary “long” D7 proteins of mosquitoes. We observed that a single JH III molecule is contained in the N-terminal domain binding pocket that is closed in an apparent conformational change by a C-terminal domain-derived α-helix. The electron density for the ligand indicated a high occupancy of the natural 10R enantiomer of JH III. Of note, mJHBP is structurally unrelated to hemolymph JHBP from lepidopteran insects. A low level of expression of mJHBP in Ae. aegypti larvae suggests that it is primarily active during the adult stage where it could potentially influence the effects of JH on egg development, mating behavior, feeding, or other processes.

A mosquito hemolymph odorant-binding protein family member specifically binds juvenile hormone.

Science.gov (United States)

Kim, Il Hwan; Pham, Van; Jablonka, Willy; Goodman, Walter G; Ribeiro, José M C; Andersen, John F

2017-09-15

Juvenile hormone (JH) is a key regulator of insect development and reproduction. In adult mosquitoes, it is essential for maturation of the ovary and normal male reproductive behavior, but how JH distribution and activity is regulated after secretion is unclear. Here, we report a new type of specific JH-binding protein, given the name mosquito juvenile hormone-binding protein (mJHBP), which circulates in the hemolymph of pupal and adult Aedes aegypti males and females. mJHBP is a member of the odorant-binding protein (OBP) family, and orthologs are present in the genomes of Aedes , Culex , and Anopheles mosquito species. Using isothermal titration calorimetry, we show that mJHBP specifically binds JH II and JH III but not eicosanoids or JH derivatives. mJHBP was crystallized in the presence of JH III and found to have a double OBP domain structure reminiscent of salivary "long" D7 proteins of mosquitoes. We observed that a single JH III molecule is contained in the N-terminal domain binding pocket that is closed in an apparent conformational change by a C-terminal domain-derived α-helix. The electron density for the ligand indicated a high occupancy of the natural 10 R enantiomer of JH III. Of note, mJHBP is structurally unrelated to hemolymph JHBP from lepidopteran insects. A low level of expression of mJHBP in Ae. aegypti larvae suggests that it is primarily active during the adult stage where it could potentially influence the effects of JH on egg development, mating behavior, feeding, or other processes.

Change of hemoagglutinin and hemolysin titers in hemolymph of gastropod molluscs in response to immunization with sheep erythrocytes

NARCIS (Netherlands)

Baskakov, AV; Polevshchikov, AV; Harazova, AD; Krasnodembskii, EG

2000-01-01

This work deals with analysis of changes of the levels of hemoagglutinins (HA) and hemolysins (HL) in hemolymph of three gastropod species, Planorbius corneus, Lymnea stagnalis, and Achatina fulica, in response to immunization with sheep erythrocytes (ShE). The levels of HA and HL were determined

Life history and environment of Cecropia latiloba in Amazonian floodplains

Directory of Open Access Journals (Sweden)

Pia Parolin

2002-06-01

Full Text Available Cecropia latiloba can be considered to be one of the most efficient colonizers of open areas in the nutrient-rich whitewater floodplains of the Amazon river. Its main strategy to be successful is the high tolerance towards waterlogging and submergence, and the fast vertical growth and reiteration capacity. This, and the tolerance of high irradiation and sediment deposition allow C. latiloba to form large monospecific stands on open sites, and thus the first closed canopy which represents the initial phase of a successional sequence which leads to highly diverse forests. This tree is extremely well adapted to the adverse growth conditions in Amazonian floodplains with prolongued periods of flooding and seedling submergence. The species occurs on the lowest levels in the flooding gradient. Although it belongs to the most often cited species under aspects of taxonomy, species distribution and general descriptions of the ecosystem, little has been published about its ecology. In the present paper the ecological, physiological and phenological characteristics of C. latiloba are described. It is an evergreen species which constantly produces new leaves. With flooding, leaf production is reduced but new leaves are flushed also with prolongued flooding. The peak of flowering and fruiting are in the flooded period. When mature, the fruits are dispersed mainly by water and fish. Seed germination occurs, without dormancy, within 5-13 days after water retreat. In the 7 months before the first flooded period seedlings reach 1 m of height, and height growth increases until a height of 15-20 m is achieved. Photosynthetic assimilation is high, with values of up to 21 mmol CO2m-2s-1 . C. latiloba is a very flood tolerant species, and waterlogged seedlings continuously produce new leaves and adventitiuos rootsCecropia latiloba puede ser considerada una de las especies colonizadoras más eficientes de áreas abiertas en las llanuras inundadas de agua dulce, rica

TOTAL HEMOCYTE COUNT AND HEMOLYMPH GLUCOSE CONCENRATION RESPONSE OF SPINY LOBSTER Panulirus homarus ON RATIO OF SHELTER

Directory of Open Access Journals (Sweden)

Suhaiba Djai

2017-11-01

Full Text Available This research was conducted to assess the physiological response of the lobster Panulirus homarus for the ratio of the shelters. The method used completely randomized design with two replicates of each treatments with shelter ratio (A 1 : 5, (B 3 : 5, (C 4 : 5, (D 5 : 5. Weight average for 184 lobsters with the stocking density of 23 lobsters for each treatment was 32.64 ± 0.58 g. The experiment was conducted for 60 days. The lobster was fed with trash fish and acclimatized for 7 days before the experiment. Observations on the physiologycal of every 10 days. The physiological responses that observed were total hemocyte count (THC and hemolymph glucose concentration. The results showed that 4:5 was the best lobster shelter ratio because it could reduce stress levels. This is indicated by the stable values of THC and hemolymph glucose level during the experiment and supported by the growth of 57.28 ± 0.15 g and survival rate of 91.31 ± 2.60%. Keywords: lobster, Panulirus homarus, ratio, shelter, THC, glucose

Milky hemolymph syndrome (MHS) in spiny lobsters, penaeid shrimp and crabs.

Science.gov (United States)

Nunan, Linda M; Poulos, Bonnie T; Navarro, Solangel; Redman, Rita M; Lightner, Donald V

2010-09-02

Black tiger shrimp Penaeus monodon, European shore crab Carcinus maenas and spiny lobster Panulirus spp. can be affected by milky hemolymph syndrome (MHS). Four rickettsia-like bacteria (RLB) isolates of MHS originating from 5 geographical areas have been identified to date. The histopathology of the disease was characterized and a multiplex PCR assay was developed for detection of the 4 bacterial isolates. The 16S rRNA gene and 16-23S rRNA intergenic spacer region (ISR) were used to examine the phylogeny of the MHS isolates. Although the pathology of this disease appears similar in the various different hosts, sequencing and examination of the phylogenetic relationships reveal 4 distinct RLB involved in the infection process.

Proteome Analysis of the Hemolymph, Mushroom Body, and Antenna Provides Novel Insight into Honeybee Resistance against Varroa Infestation.

Science.gov (United States)

Hu, Han; Bienefeld, Kaspar; Wegener, Jakob; Zautke, Fred; Hao, Yue; Feng, Mao; Han, Bin; Fang, Yu; Wubie, Abebe Jenberie; Li, Jianke

2016-08-05

Varroa destructor has been identified as a major culprit responsible for the losses of millions of honeybee colonies. Varroa sensitive hygiene (VSH) is a suite of behaviors from adult bees to suppress mite reproduction by uncapping and/or removing mite infested pupae from a sealed brood. Despite the efforts to elucidate the molecular underpinnings of VSH, they remain largely unknown. We investigated the proteome of mushroom bodies (MBs) and antennae of adult bees with and without VSH from a stock selected for VSH based on their response to artificially Varroa-infected brood cells by near-infrared camera observation. The pupal hemolymph proteome was also compared between the VSH-line and the line that was not selected for VSH. The identified 8609 proteins in the hemolymph, MBs, and antennae represent the most depth coverage of the honeybee proteome (>55%) to date. In the hemolymph, the VSH-line adapts a unique strategy to boost the social immunity and drive pupal organogenesis by enhancing energy metabolism and protein biosynthesis. In MBs, the up-regulated proteins implicated in neuronal sensitivity suggest their roles to promote the execution of VSH by activation of synaptic vesicles and calcium channel activities. In antennae, the highly expressed proteins associated with sensitivity of olfactory senses and signal transmissions signify their roles by inputting a strong signal to the MBs for initiating VSH. These observations illustrate that the enhanced social immunities and olfactory and neuronal sensitivity play key roles in the combat against Varroa infestation. The identified candidate markers may be useful for accelerating marker-associated selection for VSH to aid in resistance to a parasite responsible for decline in honeybee health.

Lipids analysis in hemolymph of African giant Achatina fulica (Bowdich, 1822) exposed to different photoperiods.

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Lustrino, D; Tunholi-Alves, V M; Tunholi, V M; Marassi, M P; Pinheiro, J

2010-02-01

The influence of different photophases (0, 6, 12, 18 and 24 hours) on the triglycerides and total cholesterol contents in the hemolymph of A. fulica was evaluated, since there is no information in the literature about the influence of this factor on lipids metabolism in mollusks. After 2 and 4 weeks of exposure the snails were dissected. The cholesterol content at the 2nd and 4th weeks post exposure only varied significantly in the groups exposed at 24 hours and 0 hour of photophase, respectively. Probably, such increase may be a result of a rise in cholesterol biosynthesis and/or remodelling of cell membranes. There were no significant differences among the content of triglycerides in the snails exposed to 6, 12, 18 and 24 hours of photophase during two weeks. The snails exposed to intermediate photophase (6 and 12 hours) had the triglycerides content increased, ranging over values near to those observed in the group exposed to 0 hour. Results showed that triglycerides metabolism in A. fulica are more influenced by photoperiod than cholesterol metabolism. A negative relation is maintained between the triglycerides content in the hemolymph and the different photophases, with lower mobilisation of triglycerides under shorter photophases.

Lipids analysis in hemolymph of African giant Achatina fulica (Bowdich, 1822 exposed to different photoperiods

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D. Lustrino

Full Text Available The influence of different photophases (0, 6, 12, 18 and 24 hours on the triglycerides and total cholesterol contents in the hemolymph of A. fulica was evaluated, since there is no information in the literature about the influence of this factor on lipids metabolism in mollusks. After 2 and 4 weeks of exposure the snails were dissected. The cholesterol content at the 2nd and 4th weeks post exposure only varied significantly in the groups exposed at 24 hours and 0 hour of photophase, respectively. Probably, such increase may be a result of a rise in cholesterol biosynthesis and/or remodelling of cell membranes. There were no significant differences among the content of triglycerides in the snails exposed to 6, 12, 18 and 24 hours of photophase during two weeks. The snails exposed to intermediate photophase (6 and 12 hours had the triglycerides content increased, ranging over values near to those observed in the group exposed to 0 hour. Results showed that triglycerides metabolism in A. fulica are more influenced by photoperiod than cholesterol metabolism. A negative relation is maintained between the triglycerides content in the hemolymph and the different photophases, with lower mobilisation of triglycerides under shorter photophases.

Thiol oxidation of hemolymph proteins in oysters Crassostrea brasiliana as markers of oxidative damage induced by urban sewage exposure.

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Trevisan, Rafael; Flores-Nunes, Fabrício; Dolores, Euler S; Mattos, Jacó J; Piazza, Clei E; Sasaki, Sílvio T; Taniguchi, Satie; Montone, Rosalinda C; Bícego, Márcia C; Dos Reis, Isis M M; Zacchi, Flávia L; Othero, Bárbara N M; Bastolla, Camila L V; Mello, Danielle F; Fraga, Ana Paula M; Wendt, Nestor; Toledo-Silva, Guilherme; Razzera, Guilherme; Dafre, Alcir L; de Melo, Cláudio M R; Bianchini, Adalto; Marques, Maria R F; Bainy, Afonso C D

2017-07-01

Urban sewage is a concerning issue worldwide, threatening both wildlife and human health. The present study investigated protein oxidation in mangrove oysters (Crassostrea brasiliana) exposed to seawater from Balneário Camboriú, an important tourist destination in Brazil that is affected by urban sewage. Oysters were exposed for 24 h to seawater collected close to the Camboriú River (CAM1) or 1 km away (CAM2). Seawater from an aquaculture laboratory was used as a reference. Local sewage input was marked by higher levels of coliforms, nitrogen, and phosphorus in seawater, as well as polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs), linear alkylbenzenes (LABs), and fecal steroid in sediments at CAM1. Exposure of oysters to CAM1 caused marked bioaccumulation of LABs and decreased PAH and PCB concentrations after exposure to both CAM1 and CAM2. Protein thiol oxidation in gills, digestive gland, and hemolymph was evaluated. Lower levels of reduced protein thiols were detected in hemolymph from CAM1, and actin, segon, and dominin were identified as targets of protein thiol oxidation. Dominin susceptibility to oxidation was confirmed in vitro by exposure to peroxides and hypochlorous acid, and 2 cysteine residues were identified as potential sites of oxidation. Overall, these data indicate that urban sewage contamination in local waters has a toxic potential and that protein thiol oxidation in hemolymph could be a useful biomarker of oxidative stress in bivalves exposed to contaminants. Environ Toxicol Chem 2017;36:1833-1845. © 2016 SETAC. © 2016 SETAC.

Determination of albendazole and metabolites in silkworm Bombyx mori hemolymph by ultrafast liquid chromatography tandem triple quadrupole mass spectrometry.

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Li, Li; Xing, Dong-Xu; Li, Qing-Rong; Xiao, Yang; Ye, Ming-Qiang; Yang, Qiong

2014-01-01

Albendazole is a broad-spectrum parasiticide with high effectiveness and low host toxicity. No method is currently available for measuring albendazole and its metabolites in silkworm hemolymph. This study describes a rapid, selective, sensitive, synchronous and reliable detection method for albendazole and its metabolites in silkworm hemolymph using ultrafast liquid chromatography tandem triple quadrupole mass spectrometry (UFLC-MS/MS). The method is liquid-liquid extraction followed by UFLC separation and quantification in an MS/MS system with positive electrospray ionization in multiple reaction monitoring mode. Precursor-to-product ion transitions were monitored at 266.100 to 234.100 for albendazole (ABZ), 282.200 to 208.100 for albendazole sulfoxide (ABZSO), 298.200 to 159.100 for albendazole sulfone (ABZSO2) and 240.200 to 133.100 for albendazole amino sulfone (ABZSO2-NH2). Calibration curves had good linearities with R2 of 0.9905-0.9972. Limits of quantitation (LOQs) were 1.32 ng/mL for ABZ, 16.67 ng/mL for ABZSO, 0.76 ng/mL for ABZSO2 and 5.94 ng/mL for ABZSO2-NH2. Recoveries were 93.12%-103.83% for ABZ, 66.51%-108.51% for ABZSO, 96.85%-105.6% for ABZSO2 and 96.46%-106.14% for ABZSO2-NH2, (RSDs albendazole and its metabolites in silkworm hemolymph in a pharmacokinetic study. The results of single-dose treatment suggested that the concentrations of ABZ, ABZSO and ABZSO2 increased and then fell, while ABZSO2-NH2 level was low without obvious change. Different trends were observed for multi-dose treatment, with concentrations of ABZSO and ABZSO2 rising over time.

Determination of Albendazole and Metabolites in Silkworm Bombyx mori Hemolymph by Ultrafast Liquid Chromatography Tandem Triple Quadrupole Mass Spectrometry

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Li, Li; Xing, Dong-Xu; Li, Qing-Rong; Xiao, Yang; Ye, Ming-Qiang; Yang, Qiong

2014-01-01

Albendazole is a broad-spectrum parasiticide with high effectiveness and low host toxicity. No method is currently available for measuring albendazole and its metabolites in silkworm hemolymph. This study describes a rapid, selective, sensitive, synchronous and reliable detection method for albendazole and its metabolites in silkworm hemolymph using ultrafast liquid chromatography tandem triple quadrupole mass spectrometry (UFLC-MS/MS). The method is liquid-liquid extraction followed by UFLC separation and quantification in an MS/MS system with positive electrospray ionization in multiple reaction monitoring mode. Precursor-to-product ion transitions were monitored at 266.100 to 234.100 for albendazole (ABZ), 282.200 to 208.100 for albendazole sulfoxide (ABZSO), 298.200 to 159.100 for albendazole sulfone (ABZSO2) and 240.200 to 133.100 for albendazole amino sulfone (ABZSO2-NH2). Calibration curves had good linearities with R2 of 0.9905–0.9972. Limits of quantitation (LOQs) were 1.32 ng/mL for ABZ, 16.67 ng/mL for ABZSO, 0.76 ng/mL for ABZSO2 and 5.94 ng/mL for ABZSO2-NH2. Recoveries were 93.12%–103.83% for ABZ, 66.51%–108.51% for ABZSO, 96.85%–105.6% for ABZSO2 and 96.46%–106.14% for ABZSO2-NH2, (RSDs <8%). Accuracy, precision and stability tests showed acceptable variation in quality control (QC) samples. This analytical method successfully determined albendazole and its metabolites in silkworm hemolymph in a pharmacokinetic study. The results of single-dose treatment suggested that the concentrations of ABZ, ABZSO and ABZSO2 increased and then fell, while ABZSO2-NH2 level was low without obvious change. Different trends were observed for multi-dose treatment, with concentrations of ABZSO and ABZSO2 rising over time. PMID:25255321

A tetrodotoxin-binding protein in the hemolymph of shore crab Hemigrapsus sanguineus: purification and properties.

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Nagashima, Yuji; Yamamoto, Kazuhiko; Shimakura, Kuniyoshi; Shiomi, Kazuo

2002-06-01

The shore crab Hemigrapsus sanguineus hemolymph contains soluble proteins that bind tetrodotoxin (TTX) and are responsible for high resistance of the crab to TTX. The TTX-binding protein was purified from the hemolymph by ultrafiltration, lectin affinity chromatography and gel filtration HPLC. The purified protein gave only one band in native-polyacrylamide gel electrophoresis (PAGE), confirming its homogeneity. Its molecular weight was estimated to be about 400k by gel filtration HPLC, while it was estimated to be about 82k under non-reducing conditions and about 72 and 82k under reducing conditions by SDS-PAGE, indicating that the TTX-binding protein was composed of at least two distinct subunits. The TTX-binding protein was an acidic glycoprotein with pI 3.5, abundant in Asp and Glu but absent in Trp, and contained 6% reducing sugar and 12% amino sugar. The protein selectively bound to TTX, with a neutralizing ability of 6.7 mouse unit TTX/mg protein, but not to paralytic shellfish poisoning toxins. However, its neutralizing activity was almost lost by treatments with enzymes (protease XIV, thermolysin, trypsin, amyloglucosidase and alpha-amylase) and denaturing agents (1% SDS, 1% dithiothreitol, 8 M urea and 6 M guanidine hydrochloride), suggesting the involvement of both proteinaceous and sugar moieties in the binding to TTX and the importance of the steric conformation of the TTX-binding protein. Copright 2002 Elsevier Science Ltd.

Review of Theoretical Prediction Models for Organic Extract Metabolites, Effect of Drying Temperature on Smooth Muscle Relaxing Activity Induced by Organic Extracts Specially Cecropia Obtusifolia Portal and Web Server Predictors of Drug-Protein Interaction.

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Aguirre-Crespo, Francisco; García-Mera, Xerardo; Guillén-Poot, Mónica Anahi; May-Díaz, Héctor Fernado; Tun-Suárez, Adrián; Aguirre-Crespo, A; Hernández-Rodríguez, J; Vergara-Galicia, Jorge; Rodríguez-López, V; Prado-Prado, Francisco J

2015-02-19

Cecropia obtusifolia bertol is medicinal specie used in the treatment of diabetes mellitus and hypertension and it has scientific studies that support the traditional use. However, it is required to understand the influence of drying temperature on the yield and pharmacological activity. Drying rate, extraction efficiency, changes in the UV-Vis spectrum and estimating chlorophylls were stimulated with the increasing temperature. Finally, relaxant activity of vascular smooth muscle is increased by 70ºC and reducing ability by the method of CARF increases with temperature. Analytical studies are required to identify changes in the metabolic content and those that ensure the safety and efficacy for human consumption. In this sense, bioinformatic studies may be helpful. Studies such as QSAR can help us to study these metabolites derived from natural products. MIND-BETS model and NL MIND-BETS model to predict DPIs was introduced using MARCH-INSIDE (MI) software to calculate structural parameters for drugs and enzymes respectively. We firstly revised the state-of-art on the design with review of previous works with hypertension activity based on theoretical studies. A study, evaluating the effect of drying temperature of leaves of C. obtusifolia on the relaxing of vascular smooth muscle, antioxidant activity and the presence of chlorophylls, with a focus on Cecropia metabolites. Last, we carried out QSAR studies using MIND-BEST and NL MIND-BEST web servers in order to understand the essential metabolites structural requirement for binding with receptors for FDA proteins.

Evaluation of Beauveria bassiana infection in the hemolymph serum proteins of the housefly, Musca domestica L. (Diptera: Muscidae).

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Mishra, Sapna; Kumar, Peeyush; Malik, Anushree

2017-11-01

Beauveria bassiana plays a prominent role in biocontrol of houseflies, Musca domestica (L.). Thus, a deeper insight into immune response of M. domestica during B. bassiana infection was warranted to assist the production of more efficient mycoinsecticides. The present study investigates changes in protein profile of M. domestica hemolymph serum post B. bassiana infection using two-dimensional difference gel electrophoresis (2D-DIGE) followed by identification of selected proteins by matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS). The non-infected or control group of flies showed an expression of 54 proteins, while M. domestica infected with B. bassiana expressed a total of 68 hemolymph serum proteins. Thirty three proteins were expressed in both groups of houseflies, whereas 35 proteins were exclusively expressed in infected flies and 21 proteins were exclusively expressed in control flies. Among the 33 proteins which were expressed in both groups of houseflies, 17 proteins showed downregulation, while16 proteins were upregulated in the infected flies compared to the non-infected ones. The results from this study are expected to facilitate better understanding of insect's immune response mechanism.

Avaliação do efeito mutagênico do extrato hidroalcoólico bruto, por meio de bioensaios in vivo e prospecção fitoquímica de Cecropia glaziovii Sneth (embaúba), Cecropiaceae

OpenAIRE

Stange, Victor S.; Gomes, Tarsila D. U. H.; Andrade, Marcieni A. de; Batitucci, Maria do Carmo P.

2009-01-01

O gênero Cecropia é conhecido popularmente por "embaúba" e apresenta várias espécies medicinais, sendo que estudos com a espécie C. glaziovii Sneth indicam que o extrato aquoso apresenta efeitos broncodilatador, anti-hipertensivo e antidepressivo, provavelmente, atribuídos às catequinas, procianidinas e flavonóides. O presente estudo objetivou avaliar os potenciais efeitos tóxico, citotóxicos, clastogênicos e aneugênicos do extrato bruto hidroalcoólico de folhas de C. glaziovii, por meio da d...

C-reactive protein in the hemolymph of Achatina fulica: interrelationship with sex steroids and metallothionein.

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Bose, R; Bhattacharya, S

2000-04-01

C-reactive protein in Achatina fulica (ACRP) is a normal component of the hemolymph. Its concentration varied from 1mg/ml in the newly hatched male, 3-5 mg/ml in the most active hermaphrodite and 1.5-2.8 mg/ml in the sedentary female showing a direct relationship of the protein with the active phase of the animal. ACRP has a molecular mass of 400 kDa and showed high absorbance in the region of 200-230 nm. It has four subunits with relative molecular masses of 110, 90, 62 and 60 kDa, respectively. Interestingly, rat platelet aggregation in vitro was significantly enhanced by ACRP in presence of 10 microM ADP and 2 mM Ca(2+) suggesting a probable role of ACRP in the aggregation of amoebocytes during the formation of plug in injured tissue. Like other vertebrate CRPs, ACRP also acts as a scavenger of chromatin fragments as evidenced by its binding to poly-L-arginine. Among the sex steroids, 4-androstenedione induces ACRP synthesis in the newly hatched male reaching the level found in the most active hermaphrodite phase (4 mg/ml). A very high molar ratio (5) of mercury binding to ACRP confirmed its sequestration property of heavy metals as observed in vertebrates. The level of metallothionein (MT) in the hemolymph gradually increased from the male to the hermaphrodite to the female, a pattern distinctly different from that of the ACRP titer. Since both MT and ACRP can sequester inorganic mercury, the high level of MT compensates functionally for the low titer of ACRP in the sedentary female.

Hemolymph Melanization in the Silkmoth Bombyx mori Involves Formation of a High Molecular Mass Complex That Metabolizes Tyrosine*

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Clark, Kevin D.; Strand, Michael R.

2013-01-01

The phenoloxidase (PO) cascade regulates the melanization of blood (hemolymph) in insects and other arthropods. Most studies indicate that microbial elicitors activate the PO cascade, which results in processing of the zymogen PPO to PO. PO is then thought to oxidize tyrosine and o-diphenols to quinones, which leads to melanin. However, different lines of investigation raise questions as to whether these views are fully correct. Here we report that hemolymph from the silkmoth, Bombyx mori, rapidly melanizes after collection from a wound site. Prior studies indicated that in vitro activated PPO hydroxylates Tyr inefficiently. Measurement of in vivo substrate titers, however, suggested that Tyr was the only PO substrate initially present in B. mori plasma and that it is rapidly metabolized by PO. Fractionation of plasma by gel filtration chromatography followed by bioassays indicated that melanization activity was primarily associated with a high mass complex (∼670 kDa) that contained PO. The prophenoloxidase-activating protease inhibitor Egf1.0 blocked formation of this complex and Tyr metabolism, but the addition of phenylthiourea to plasma before fractionation enhanced complex formation and Tyr metabolism. Mass spectrometry analysis indicated that the complex contained PO plus other proteins. Taken together, our results indicate that wounding alone activates the PO cascade in B. mori. They also suggest that complex formation is required for efficient use of Tyr as a substrate. PMID:23553628

Elemental content changes in hemolymph of Rhodnius prolixus due to mercury contamination: a study using SR-TXRF

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Mantuano, A.; Oliveira, A.P.; Barroso, R.C. [Universidade do Estado do Rio de Janeiro (UERJ), Rio de Janeiro, RJ (Brazil); Almeida, A.P.; Braz, D.; Cardoso, S.C. [Universidade Federal do Rio de Janeiro (UFRJ), Rio de Janeiro, RJ (Brazil); Figueiredo, M.B.; Azambuja, P. [Fundacao Instituto Osvaldo Cruz (FIOCRUZ), Rio de Janeiro, RJ (Brazil); Gonzalez, M.S. [Universidade Federal Fluminense (UFF), Niteroi, RJ (Brazil)

2012-07-01

Full text: In recent years, the effects of pollution on the health of humans and other vertebrates have been extensively studied. However, the effects on some invertebrates are comparatively unknown. Research has demonstrated that toxic metals interfere with the reproduction, development and immune defenses of some terrestrial and marine invertebrates. Depending on environmental conditions, pollution produces chronic and acute effects on different systems and organs of animals. In general, some more toxic elements like arsenic, cadmium, lead, mercury and nickel in contact with organisms change cellular structures, enzyme activities, and in some cases destroy the physiological integrities of the tissues. In insects, the effects of pollutants depend upon the species studied. In this work, we investigated the changes in elemental contents in the hemolymph of Rhodnius prolixus on 2 and 5 days after feeding on blood containing mercury chloride. R. Prolixus is an obligated hematophagous Hemiptera and one of the most important insect vectors of trypanosoma cruzi, the causative agent of Chagas disease. The SR-TXRF measurements were performed at the X-ray fluorescence (XRF) beamline facility in Brazilian Synchrotron Light Laboratory LNLS/Brazil. The major elements Cl, K, Ca, Mn, Fe, Cu, Zn, Se, Br, Rb, K were found in all hemolymph samples analysed. Insects treated with HgCl2 had reduced Cl and Ca levels, whereas the same treatment had enhanced Br levels in comparison with non- treated control insects. (author)

Nest structure and occurrence of three species of Azteca (Hymenoptera, Formicidae in Cecropia pachystachya (Urticaceae in non-floodable and floodable pantanal areas Arquitetura de ninho e ocorrência de três espécies de Azteca (Hymenoptera, Formicidae em Cecropia pachystachya (Urticaceae em ambiente alagável e não alagável no Pantanal

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Alexsandro S. Vieira

2010-01-01

Full Text Available Thirty Cecropia pachystachya trees were examined in non-floodable and floodable areas to investigate the association between C. pachystachya and Azteca ants in the Pantanal of Mato Grosso do Sul, Brazil. The species Azteca ovaticeps, Azteca isthmica, and Azteca alfari were found nesting inside domatia of C. pachystachya. A. ovaticeps was the most frequent species in the trees in the floodable area, while A. isthmica and A. alfari, in the non-floodable area. A. ovaticeps and A. isthmica maintained more entrance/exit holes in comparison to A. alfari. All Azteca species maintained entrance/exit holes in the closest domatia to the apical area of the branch, due to proximity to Müllerian and pearl bodies, suggesting that these species of Azteca were influenced by their environment during evolution and became specialized. All internodal septa of each examined branch were perforated by ants, indicating the branches were inhabited by a single colony.Foram analisadas 30 plantas de Cecropia pachystachya em cada ambiente alagável e não alagável no Pantanal sul-mato-grossense, Brasil, com o objetivo de investigar a associação entre formigas Azteca e C. pachystachya. Foram encontradas as espécies Azteca ovaticeps, Azteca isthmica e Azteca alfari nidificando nas domáceas da planta. A. ovaticeps foi mais frequente em plantas de área alagável, enquanto A. isthmica e A. alfari em plantas em área não alagável. A. ovaticeps e A. isthmica apresentaram maior quantidade de orifícios de entrada/saída em relação à A. alfari e todas as espécies mantêm próximo da região apical do ramo, orifícios de entrada/saída nas domáceas, devido à proximidade com os corpúsculos müellerianos e pérola. Isto é, essas espécies de Azteca especializaram-se ao longo da evolução influenciadas pelo ambientes. Todos os septos internodais de cada ramo analisados apresentaram-se perfurados pelas formigas, sugerindo que os mesmos são habitados por uma única colônia.

PHOSPHATE AND INOCULATION WITH ARBUSCULAR MYCORRHIZAL FUNGI ON THE GROWTH OF Cecropia pachystachya (Trec SEEDLINGS FÓSFORO E INOCULAÇÃO COM FUNGOS MICORRÍZICOS ARBUSCULARES NO ESTABELECIMENTO DE MUDAS DE EMBAÚBA (Cecropia pachystachya Trec

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Marco Aurélio Carbone Carneiro

2007-09-01

Full Text Available

The objective of this study was to evaluate the effect of the inoculation with arbuscular mycorrhizal fungi (FMA in different levels of P₂O₅ on the growth of Cecropia pachystachya seedlings in the field. The study consisted of a 5x2 factorial with five levels of P₂O₅ (zero, 85, 170, 255 and 340 mg.kg^-1, with and ithout inoculation with a mixture of FMA. It was used four replications, each one with twelve seedlings. The seeds were sowed in plastic tubes with capacity of 50 cm³ of substratum and stored for 120 days. After this period the seedlings were transplanted to the field, where they remained for another 150 days. Seedling diameter and height were measured at 60 and 120 days, aerial part and root dry matter, and arbuscular mycorrhizal colonization. Diameter, height, leaf area, aerial part dry matter and the number of surviving seedlings were determined after 150 days. None of the factors tested had any effect on seedling growth with one exception; inoculated plants with FMA had more root dry matter. Plants inoculated with smaller doses of P₂O₅ showed a larger percentage of surviving individuals and more vigorous seedlings. Results suggest that in low fertility soils of and subject to the hydric stress the C. pachystachya seedlings should be inoculated with FMA.

KEY-WORDS: Native vegetation; mycorrhiza fungi; native species; seedling production; inoculation.

O objetivo deste estudo foi avaliar o efeito da inoculação com fungos micorrízicos arbusculares (FMA em diferentes doses de P₂O₅ na formação de mudas de embaúba (Cecropia pachystachya e no seu estabelecimento em campo. O estudo constou de um experimento fatorial 5x2, sendo cinco doses de P₂O₅

Evolutionary morphology of the hemolymph vascular system of basal araneomorph spiders (Araneae: Araneomorphae).

Science.gov (United States)

Huckstorf, Katarina; Michalik, Peter; Ramírez, Martín; Wirkner, Christian S

2015-11-01

The superfamily Austrochiloidea (Austrochilidae and Gradungulidae) take a pivotal position in araneomorph spider phylogeny. In this discussion crevice weaver spiders (Filistatidae) are of equal interest. Especially data from these phylogenetically uncertain yet basal off branching groups can enlighten our understanding on the evolution of organ systems. In the course of a survey on the evolutionary morphology of the circulatory system in spiders we therefore investigated the hemolymph vascular system in two austrochiloid and one filistatid species. Additionally some data on a hypochilid and a gradungulid species are included. Using up-to-date morphological methods, the vascular systems in these spiders are visualized three dimensionally. Ground pattern features of the circulatory systems in austrochiloid spiders are presented and the data discussed along recent lines of phylogenetic hypotheses. Special topics highlighted are the intraspecific variability of the origins of some prosomal arteries and the evolutionary correlation of respiratory and circulatory systems in spiders. Copyright © 2015 Elsevier Ltd. All rights reserved.

The effect of symbiotic ant colonies on plant growth: a test using an Azteca-Cecropia system.

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Karla N Oliveira

Full Text Available In studies of ant-plant mutualisms, the role that ants play in increasing the growth rates of their plant partners is potentially a key beneficial service. In the field, we measured the growth of Cecropia glaziovii saplings and compared individuals that were naturally colonized by Azteca muelleri ants with uncolonized plants in different seasons (wet and dry. We also measured light availability as well as attributes that could be influenced by the presence of Azteca colonies, such as herbivory, leaf nutrients (total nitrogen and δ(15N, and investments in defense (total phenolics and leaf mass per area. We found that colonized plants grew faster than uncolonized plants and experienced a lower level of herbivory in both the wet and dry seasons. Colonized plants had higher nitrogen content than uncolonized plants, although the δ(15N, light environment, total phenolics and leaf mass per area, did not differ between colonized and uncolonized plants. Since colonized and uncolonized plants did not differ in the direct defenses that we evaluated, yet herbivory was lower in colonized plants, we conclude that biotic defenses were the most effective protection against herbivores in our system. This result supports the hypothesis that protection provided by ants is an important factor promoting plant growth. Since C. glaziovii is widely distributed among a variety of forests and ecotones, and since we demonstrated a strong relationship with their ant partners, this system can be useful for comparative studies of ant-plant interactions in different habitats. Also, given this study was carried out near the transition to the subtropics, these results help generalize the geographic distribution of this mutualism and may shed light on the persistence of the interactions in the face of climate change.

Binding specificity of the juvenile hormone carrier protein from the hemolymph of the tobacco hornworm Manduca sexta Johannson (Lepidoptera: Sphingidae).

Science.gov (United States)

Peterson, R C; Reich, M F; Dunn, P E; Law, J H; Katzenellnbogen, J A

1977-05-17

A series of analogues of insect juvenile hormone (four geometric isomers of methyl epoxyfarnesenate, several para-substituted epoxygeranyl phenyl ethers, and epoxyfarnesol and its acetate and haloacetate derivatives) was prepared to investigate the binding specificity of the hemolymph juvenile hormone binding protein from the tobacco hornworm Manduct sexta. The relative binding affinities were determined by a competition assay against radiolabeled methyl (E,E)-3,11-dimethyl-7-ethyl-cis-10,11-epoxytrideca-2,6-dienoate (JH I). The ratio of dissociation constants was estimated by plotting competitor data according to a linear transformation of the dissociation equations describing competition of two ligands for a binding protein. The importance of the geometry of the sesquiterpene hydrocarbon chain is indicated by the fact that the binding affinity is decreased as Z (cis) double bonds are substituted for E (trans) double bonds in the methyl epoxyfarnesenate series; the unepoxidized analogues do not bind. A carboxylic ester function is important although its orientation can be reversed, as indicated by the good binding of epoxyfarnesyl acetate. In the monoterpene series, methyl epoxygeranoate shows no affinity for the binding protein, but substitution of a phenyl or p-carbomethoxyphenyl ether for the ester function imparts a low, but significant affinity. These data taken together with earlier results indicate that the binding site for juvenile hormone in the hemolymph binding protein is characterized by a sterically defined hydrophobic region with polar sites that recognize the epoxide and the ester functions.

Isolation, cDNA cloning, and structure-based functional characterization of oryctin, a hemolymph protein from the coconut rhinoceros beetle, Oryctes rhinoceros, as a novel serine protease inhibitor.

Science.gov (United States)

Horita, Shoichiro; Ishibashi, Jun; Nagata, Koji; Miyakawa, Takuya; Yamakawa, Minoru; Tanokura, Masaru

2010-09-24

We isolated oryctin, a 66-residue peptide, from the hemolymph of the coconut rhinoceros beetle Oryctes rhinoceros and cloned its cDNA. Oryctin is dissimilar to any other known peptides in amino acid sequence, and its function has been unknown. To reveal that function, we determined the solution structure of recombinant (13)C,(15)N-labeled oryctin by heteronuclear NMR spectroscopy. Oryctin exhibits a fold similar to that of Kazal-type serine protease inhibitors but has a unique additional C-terminal α-helix. We performed protease inhibition assays of oryctin against several bacterial and eukaryotic proteases. Oryctin does inhibit the following serine proteases: α-chymotrypsin, endopeptidase K, subtilisin Carlsberg, and leukocyte elastase, with K(i) values of 3.9 × 10(-10) m, 6.2 × 10(-10) m, 1.4 × 10(-9) m, and 1.2 × 10(-8) m, respectively. Although the target molecule of oryctin in the beetle hemolymph remains obscure, our results showed that oryctin is a novel single domain Kazal-type inhibitor and could play a key role in protecting against bacterial infections.

Tissue culture of Cecropia glaziovii Sneth (urticaceae: vegetative micropropagation and plant regeneration from callus Cultura de tecidos de Cecropia glaziovii Sneth (Urticaceae: micropropagação vegetativa e regeneração de plantas via calos

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Marcos Nopper Alves

2010-10-01

Full Text Available Cecropia glaziovii is a tree with used in Brazilian popular medicine. Methods allowing the clonal propagation of this species are of great interest for superior genotype multiplication and perpetuation. For this reason, we examined the effect of different culture media and different types of explants on adventitious shoot regeneration from callus and buds of C. glaziovii. Leaves, petioles and stipules obtained from aseptically grown seedlings or from pre-sterilized plants were used to initiate cultures. Adventitious shoot regeneration was achieved when apical and axillary buds were inoculated on gelled Murashige & Skoog (MS medium supplemented with 6-benzylaminopurine alone (BAP (1.0, 5.0 or 10.0 mg L-1 or combined with -naphthalene acetic acid (NAA (1.0 or 2.0 mg L-1, after 40 days of culture. Best callus production was obtained after 30 days of petioles' culture on gelled MS medium with 2,4 dichlorophenoxyacetic acid (2,4-D (5.0 mg L-1 combined with BAP (1.0 mg L-1. Successful shoot regeneration from callus was achieved when MS medium supplemented with zeatin (ZEA (0.1 mg L-1 alone or combined with 2,4-D (1.0 or 5.0 mg L-1 was inoculated with friable callus obtained from petioles. All shoots were rooted by inoculation on MS medium supplemented with indole-3-acetic acid (IAA (1.0 mg L-1. Rooted plants transferred to potting soil were successfully established. All in vitro regenerated plantlets showed to be normal, without morphological variations, being also identical to the source plant. Our study has shown that C. glaziovii can be propagated by tissue culture methods, allowing large scale multiplication of superior plants for pharmacological purposes.Cecropia glaziovii é uma planta lenhosa, popularmente usada no Brasil como medicinal. Métodos que visem a sua propagação clonal podem ser de grande utilidade na preservação de seus genótipos de elite. Foram examinados efeitos de diferentes reguladores de crescimento e explantes na forma

Ação da inoculação de hemolinfa no mecanismo de defesa de Biomphalaria tenagophila (Orbigny, 1835 The effect of hemolymph inoculation on the defense mechanism of Biomphalaria tenagophila (Orbigny, 1835

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Soely Maria Pissini Machado Reis

1995-08-01

Full Text Available Estudou-se a resistência à infecção pelo S. mansoni em moluscos B. tenagophila inoculados com vários tipos de hemolinfa provenientes de moluscos infectados por S. mansoni da linhagem SJ ou por outros trematódeos avaliando-se, nestas circunstâncias, o comportamento dos amebócitos. Concluiu-se que dois tipos de mecanismos teriam agido provocando resistência à infecção pelo S. mansoni: celular e humoral. A reação do tipo celular ocorreu quando os moluscos eram inoculados com hemolinfa de moluscos infectados por S. mansoni B. tenagophila inoculadas com hemolinfa de moluscos infectados por furcocercárias longifurcadas sem ocelos, apresentaram elevada resistência à infecção posterior por S.mansoni.The resistance of B. tenagophila snails to infection by Schistosoma mansoni was studied. These snails had been previously inoculated with hemolymph talsen from other snails infected by S. mansoni of SJ strain or by other trematodes. The findings suggest that two types - cellular and humoral - are responsible for the resistance to infection by S. mansoni. The cellular response occurred when the snails were inoculated with hemolymph from snails infected by S. mansoni. B. tenagophila inoculate with hemolymph from snails infected by furcocercariae without eye-spot, also showed resistance to the infection by S. mansoni. In this latter case, the degenerated sporocysts were not enveloped by amebocitary reaction, leading to the conclusion that humoral factors could be involved.

Free amino acid pools in muscle and hemolymph during the molt cycle of the land crab, Gecarcinus lateralis

Energy Technology Data Exchange (ETDEWEB)

Yamaoka, L.H.; Skinner, D.M.

1976-01-01

The total free amino acids in muscle water of the land crab, Gecarcinus lateralis, decrease almost 3-fold during the premolt period in comparison to the intermolt period (193 ..mu..M/g during intermolt and 67 ..mu..M/g during late premolt). This decrease is accounted for primarily by changes in the nonessential amino acids proline, glycine and alanine. At the same stage, several essential amino acids (lysine, methionine and tyrosine) increase 2- to 4-fold. Although free amino acid levels in hemolymph are lower and more variable than those in muscle, the same amino acids show similar molt-stage-related changes. The decreases in glycine and proline may be associated with the synthesis of the new exoskeleton and connective tissue during the premolt period.

Isolation, cDNA Cloning, and Structure-based Functional Characterization of Oryctin, a Hemolymph Protein from the Coconut Rhinoceros Beetle, Oryctes rhinoceros, as a Novel Serine Protease Inhibitor*

Science.gov (United States)

Horita, Shoichiro; Ishibashi, Jun; Nagata, Koji; Miyakawa, Takuya; Yamakawa, Minoru; Tanokura, Masaru

2010-01-01

We isolated oryctin, a 66-residue peptide, from the hemolymph of the coconut rhinoceros beetle Oryctes rhinoceros and cloned its cDNA. Oryctin is dissimilar to any other known peptides in amino acid sequence, and its function has been unknown. To reveal that function, we determined the solution structure of recombinant 13C,15N-labeled oryctin by heteronuclear NMR spectroscopy. Oryctin exhibits a fold similar to that of Kazal-type serine protease inhibitors but has a unique additional C-terminal α-helix. We performed protease inhibition assays of oryctin against several bacterial and eukaryotic proteases. Oryctin does inhibit the following serine proteases: α-chymotrypsin, endopeptidase K, subtilisin Carlsberg, and leukocyte elastase, with Ki values of 3.9 × 10−10 m, 6.2 × 10−10 m, 1.4 × 10−9 m, and 1.2 × 10−8 m, respectively. Although the target molecule of oryctin in the beetle hemolymph remains obscure, our results showed that oryctin is a novel single domain Kazal-type inhibitor and could play a key role in protecting against bacterial infections. PMID:20630859

Late rise in hemolymph osmolality in Macrobrachium acanthurus (diadromous freshwater shrimp) exposed to brackish water: Early reduction in branchial Na+/K+ pump activity but stable muscle HSP70 expression.

Science.gov (United States)

Freire, Carolina A; Maraschi, Anieli C; Lara, Alessandra F; Amado, Enelise M; Prodocimo, Viviane

2018-02-01

Some Macrobrachium shrimps (Caridea, Palaemonidae) are diadromous; freshwater adults are truly euryhaline, while larvae need saline water for development. Branchial Na + /K + -ATPase (NKA) and carbonic anhydrase (CA) are involved in NaCl absorption in freshwater. This study aimed at verifying the time course of the osmoregulatory response of adult Macrobrachium acanthurus to high salinity brackish water (20‰), from the first 30min to 5days. The goal was to detect possible transition from hyper- to hyporegulation, the putative involvement of branchial NKA and CA, or the induction of muscular HSP70 expression. Hemolymph osmotic and ionic concentrations remained relatively stable and close to control levels until ~9h of exposure, but later increased consistently (~50%). A fast reduction in NKA activity (3-6h) was observed; these shrimps seem to shut off salt absorption already in the first hours. Later on, especially after 24h, hemolymph concentrations rise but HSP70 expression is not induced, possibly because constitutive levels are already sufficient to prevent protein damage. Time-dependent response mechanisms effective in high salinity brackish water, resulting in salt loading avoidance and suggestive of hyporegulation should be further investigated in decapods that evolutionary invaded freshwater. Copyright © 2017 Elsevier Inc. All rights reserved.

Metabolic Post-feeding Changes in Fat Body and Hemolymph of Dipetalogaster maximus (Hemiptera:Reduviidae

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Lilián E Canavoso

1998-03-01

Full Text Available Lipids and glycogen in fat body as well as the modifications in the wet weight of this organ were evaluated in an unfed insect, Dipetalogaster maximus, on day 5 after adult ecdysis (time 0 and during a 30-day period after ingestion of blood meal. Total lipids, high density lipophorin (HDLp, carbohydrates, total proteins and uric acid were determined in the hemolymph during the same period. Fat body wet weight was maximum on day 10 post-feeding and represented on day 30 only 42% of the maximum weight. Lipids stored in the fat body increased up to day 15 reaching 24% of the total weight of tissue. Glycogen was maximum on day 20, representing approximately 3% of the fat body weight. HDLp represented at all times between 17-24% of the total proteins, whose levels ranged between 35 and 47 mg/ml. Uric acid showed at 20, 25 and 30 days similar levels and significantly higher than the ones shown at days 10 and 15. Hemolymphatic lipids fluctuated during starvation between 3-4.4 mg/ml and carbohydrates showed a maximum on day 15 after a blood meal, decreasing up to 0.26 mg/ml on day 25. The above results suggest that during physiological events such as starvation, the availability of nutrients is affected, involving principally the fat body reserves

Elemental changes in hemolymph and urine of Rhodnius prolixus induced by in-vitro exposure to mercury: A study using SR-TXRF

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Mantuano, Andrea; Barroso, Regina C. [Physics Institute/State University of Rio de Janeiro (UERJ), RJ (Brazil); Almeida, Andre P. de; Braz, Delson [Nuclear Engineering Program/COPPE/Federal University of Rio de Janeiro, RJ (Brazil); Cardoso, Simone C. [Physics Institute/Federal University of Rio de Janeiro, RJ (Brazil); Penna, Patricia A.; Gonzalez, Marcelo S. [Laboratory of Biochemistry and Physiology of Insects/Oswaldo Cruz Foundation (FIOCRUZ), Rio de Janeiro, RJ (Brazil)

2011-07-01

Full text: In the last years have been studied the effects of the pollution of humans and others vertebrates, however, the effects on invertebrates are poorly knows. Some pollutants introduced in aquatic and terrestrial ecosystems are potentially toxic to living organisms. Almost the environmental pollutants, the heavy metals are not degradable and can persist during long time of many ecosystems causing ecologic changes often disastrous to species that habit there. Actually some works has shown that the heavy metals beyond be toxic and interfere on development and reproduction of some species of terrestrial and marine invertebrates. When are present in cells, the chemical form and connection type of metals are critical factors that determinate the toxicity. So, the ambient pollution has chronic effects and acute of animals health and can affect any systems and organs. The intoxications that current often are caused by aluminum, arsenic, barium, beryllium, cadmium, lead, mercury and nickel. Know that these elements can change cellular structures, enzymes and replace metal cofactors in enzyme activities. In insects, the effects of pollution change according to specie studied. The pollution can act on the weight reduction and increasing the relative growth rate. In this work, we investigated the effect of mercury exposure on the elemental content in hemolymph and urine of Rhodnius prolixus the insect vector of Chagas' disease, which is one of the most important vectors in Latin American and also, the most well-know studied insect in terms of both physiology and vector-parasite interactions. Five-stage nymphs of Rhodnius prolixus were collected from colony of a Laboratory Physiology and Biochemistry of Insects, Institute Oswaldo Cruz RJ. For treatment of insects, mercury chloride has been added to rabbit blood. After feeding the nymphs were separated and packed for two days for collection of hemolymph and urine. The SR-TXRF measurements were performed at the X

Elemental changes in hemolymph and urine of Rhodnius prolixus induced by in-vitro exposure to mercury: A study using SR-TXRF

International Nuclear Information System (INIS)

Mantuano, Andrea; Barroso, Regina C.; Almeida, Andre P. de; Braz, Delson; Cardoso, Simone C.; Penna, Patricia A.; Gonzalez, Marcelo S.

2011-01-01

Full text: In the last years have been studied the effects of the pollution of humans and others vertebrates, however, the effects on invertebrates are poorly knows. Some pollutants introduced in aquatic and terrestrial ecosystems are potentially toxic to living organisms. Almost the environmental pollutants, the heavy metals are not degradable and can persist during long time of many ecosystems causing ecologic changes often disastrous to species that habit there. Actually some works has shown that the heavy metals beyond be toxic and interfere on development and reproduction of some species of terrestrial and marine invertebrates. When are present in cells, the chemical form and connection type of metals are critical factors that determinate the toxicity. So, the ambient pollution has chronic effects and acute of animals health and can affect any systems and organs. The intoxications that current often are caused by aluminum, arsenic, barium, beryllium, cadmium, lead, mercury and nickel. Know that these elements can change cellular structures, enzymes and replace metal cofactors in enzyme activities. In insects, the effects of pollution change according to specie studied. The pollution can act on the weight reduction and increasing the relative growth rate. In this work, we investigated the effect of mercury exposure on the elemental content in hemolymph and urine of Rhodnius prolixus the insect vector of Chagas' disease, which is one of the most important vectors in Latin American and also, the most well-know studied insect in terms of both physiology and vector-parasite interactions. Five-stage nymphs of Rhodnius prolixus were collected from colony of a Laboratory Physiology and Biochemistry of Insects, Institute Oswaldo Cruz RJ. For treatment of insects, mercury chloride has been added to rabbit blood. After feeding the nymphs were separated and packed for two days for collection of hemolymph and urine. The SR-TXRF measurements were performed at the X

Interactions of cationic polystyrene nanoparticles with marine bivalve hemocytes in a physiological environment: Role of soluble hemolymph proteins.

Science.gov (United States)

Canesi, Laura; Ciacci, Caterina; Fabbri, Rita; Balbi, Teresa; Salis, Annalisa; Damonte, Gianluca; Cortese, Katia; Caratto, Valentina; Monopoli, Marco P; Dawson, Kenneth; Bergami, Elisa; Corsi, Ilaria

2016-10-01

The bivalve Mytilus galloprovincialis has proven as a suitable model invertebrate for evaluating the potential impact of nanoparticles (NPs) in the marine environment. In particular, in mussels, the immune system represents a sensitive target for different types of NPs. In environmental conditions, both NP intrinsic properties and those of the receiving medium will affect particle behavior and consequent bioavailability/uptake/toxicity. However, the evaluation of the biological effects of NPs requires additional understanding of how, once within the organism, NPs interact at the molecular level with cells in a physiological environment. In mammalian systems, different NPs associate with serum soluble components, organized into a "protein corona", which affects particle interactions with target cells. However, no information is available so far on the interactions of NPs with biological fluids of aquatic organisms. In this work, the influence of hemolymph serum (HS) on the in vitro effects of amino modified polystyrene NPs (PS-NH2) on Mytilus hemocytes was investigated. Hemocytes were incubated with PS-NH2 suspensions in HS (1, 5 and 50µg/mL) and the results were compared with those obtained in ASW medium. Cell functional parameters (lysosomal membrane stability, oxyradical production, phagocytosis) were evaluated, and morphological changes were investigated by TEM. The activation state of the signalling components involved in Mytilus immune response (p38 MAPK and PKC) was determined. The results show that in the presence of HS, PS-NH2 increased cellular damage and ROS production with respect to ASW medium. The effects were apparently mediated by disregulation of p38 MAPK signalling. The formation of a PS-NH2-protein corona in HS was investigated by centrifugation, and 1D- gel electrophoresis and nano-HPLC-ESI-MS/MS. The results identified the Putative C1q domain containing protein (MgC1q6) as the only component of the PS-NH2 hard protein corona in Mytilus

Crecimiento y mortalidad en juveniles de siete especies arbóreas en un bosque muy húmedo tropical intervenido de Costa Rica

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Grace Sáenz

1999-06-01

Full Text Available Se estudiaron por un período de cuatro años los patrones de crecimiento en diámetro y altura, así como la mortalidad de brinzales (individuos e 0.30 m altura y Diameter and height growth patterns, as well as the incidence of seedling (individuals e0.3 m and <1.5 m in height and sapling (individual e1.5 m in height and 9.9 cm DBH mortality were studied for four years in a forest under two types of silvicultural intervention. The study is based on anual measurements of individuals from the two size categories marked one year after harvest. In the case of the seedlings, the parameters evaluated were: total height and diameter at 10 cm from the base. For the sapling category, the diameter at 1.30 m was also measured. The Clark and Clark methodology was used to evaluate the crown illumination index of all individuals. Seven tree species were studied: Cecropia (two species, Cecropia insignis Liebm. and Cecropia obtusifolia Bertol., Laetia procera (Poepp. Eichler, Rollinia pittieri Saff, Apeiba membranacea Spruce ex Benth., Virola koschnyi Warb. y Virola sebifera Aubl. The Cecropia spp. seedlings showed the highest diameter and height growth rates, with annual mean increments in diameter of 7.8 mm/years and height of 188 cm/years. The L. procera saplings had the greatest diameter and height growth, with rates of 7.0 mm/year and 86 cm/year, respectively. Only in the case of Cecropia spp. and V. sebifera are diameter and height growth significantly correlated with the crown light index. Seedlings had the highest mortality rates in all species except Cecropia spp. and R. pittieri, where the sapling mortality was greater.

Consumption of Mullerian Bodies by Golden-olive Woodpecker (Colaptes rubiginosus) in Nicaragua's Highlands

Science.gov (United States)

M.A. Torrez; Wayne Arendt; L. Diaz

2016-01-01

The Golden-olive Woodpecker is a generalist species found in a wide range of habitats, being particularly common in coffee plantations within Nicaraguan cloud forests. Observations of an individual feeding at the base of Cecropia leaves revealed it was consuming Mullerian bodies that the Cecropia produces to feed Azteca ants as part of a host-inhabitant mutualistic...

Interactions of cationic polystyrene nanoparticles with marine bivalve hemocytes in a physiological environment: Role of soluble hemolymph proteins

Energy Technology Data Exchange (ETDEWEB)

Canesi, Laura, E-mail: Laura.Canesi@unige.it [Dept. of Earth, Environmental and Life Sciences – DISTAV, University of Genoa (Italy); Ciacci, Caterina [Dept. of Biomolecular Sciences – DIBS, University of Urbino (Italy); Fabbri, Rita; Balbi, Teresa [Dept. of Earth, Environmental and Life Sciences – DISTAV, University of Genoa (Italy); Salis, Annalisa; Damonte, Gianluca [Centre of Excellence for Biomedical Research – CEBR, University of Genoa (Italy); Cortese, Katia [Department of Experimental Medicine – DIMES, University of Genoa (Italy); Caratto, Valentina [Dept. of Earth, Environmental and Life Sciences – DISTAV, University of Genoa (Italy); Monopoli, Marco P. [Centre for BioNanoInteractions, School of Chemistry and Chemical Biology, University College Dublin (Ireland); Department of Pharmaceutical and Medical Chemistry, Royal College of Surgeons, 123 St. Stephen Green, Dublin (Ireland); Dawson, Kenneth [Centre for BioNanoInteractions, School of Chemistry and Chemical Biology, University College Dublin (Ireland); Bergami, Elisa; Corsi, Ilaria [Dept. of Physical, Earth and Environmental Sciences, University of Siena (Italy)

2016-10-15

The bivalve Mytilus galloprovincialis has proven as a suitable model invertebrate for evaluating the potential impact of nanoparticles (NPs) in the marine environment. In particular, in mussels, the immune system represents a sensitive target for different types of NPs. In environmental conditions, both NP intrinsic properties and those of the receiving medium will affect particle behavior and consequent bioavailability/uptake/toxicity. However, the evaluation of the biological effects of NPs requires additional understanding of how, once within the organism, NPs interact at the molecular level with cells in a physiological environment. In mammalian systems, different NPs associate with serum soluble components, organized into a “protein corona”, which affects particle interactions with target cells. However, no information is available so far on the interactions of NPs with biological fluids of aquatic organisms. In this work, the influence of hemolymph serum (HS) on the in vitro effects of amino modified polystyrene NPs (PS-NH{sub 2}) on Mytilus hemocytes was investigated. Hemocytes were incubated with PS-NH{sub 2} suspensions in HS (1, 5 and 50 µg/mL) and the results were compared with those obtained in ASW medium. Cell functional parameters (lysosomal membrane stability, oxyradical production, phagocytosis) were evaluated, and morphological changes were investigated by TEM. The activation state of the signalling components involved in Mytilus immune response (p38 MAPK and PKC) was determined. The results show that in the presence of HS, PS-NH{sub 2} increased cellular damage and ROS production with respect to ASW medium. The effects were apparently mediated by disregulation of p38 MAPK signalling. The formation of a PS-NH{sub 2}-protein corona in HS was investigated by centrifugation, and 1D- gel electrophoresis and nano-HPLC-ESI-MS/MS. The results identified the Putative C1q domain containing protein (MgC1q6) as the only component of the PS-NH{sub 2} hard

Interactions of cationic polystyrene nanoparticles with marine bivalve hemocytes in a physiological environment: Role of soluble hemolymph proteins

International Nuclear Information System (INIS)

Canesi, Laura; Ciacci, Caterina; Fabbri, Rita; Balbi, Teresa; Salis, Annalisa; Damonte, Gianluca; Cortese, Katia; Caratto, Valentina; Monopoli, Marco P.; Dawson, Kenneth; Bergami, Elisa; Corsi, Ilaria

2016-01-01

The bivalve Mytilus galloprovincialis has proven as a suitable model invertebrate for evaluating the potential impact of nanoparticles (NPs) in the marine environment. In particular, in mussels, the immune system represents a sensitive target for different types of NPs. In environmental conditions, both NP intrinsic properties and those of the receiving medium will affect particle behavior and consequent bioavailability/uptake/toxicity. However, the evaluation of the biological effects of NPs requires additional understanding of how, once within the organism, NPs interact at the molecular level with cells in a physiological environment. In mammalian systems, different NPs associate with serum soluble components, organized into a “protein corona”, which affects particle interactions with target cells. However, no information is available so far on the interactions of NPs with biological fluids of aquatic organisms. In this work, the influence of hemolymph serum (HS) on the in vitro effects of amino modified polystyrene NPs (PS-NH 2 ) on Mytilus hemocytes was investigated. Hemocytes were incubated with PS-NH 2 suspensions in HS (1, 5 and 50 µg/mL) and the results were compared with those obtained in ASW medium. Cell functional parameters (lysosomal membrane stability, oxyradical production, phagocytosis) were evaluated, and morphological changes were investigated by TEM. The activation state of the signalling components involved in Mytilus immune response (p38 MAPK and PKC) was determined. The results show that in the presence of HS, PS-NH 2 increased cellular damage and ROS production with respect to ASW medium. The effects were apparently mediated by disregulation of p38 MAPK signalling. The formation of a PS-NH 2 -protein corona in HS was investigated by centrifugation, and 1D- gel electrophoresis and nano-HPLC-ESI-MS/MS. The results identified the Putative C1q domain containing protein (MgC1q6) as the only component of the PS-NH 2 hard protein corona in

Biomass-derived functional porous carbons as novel electrode material for the practical detection of biomolecules in human serum and snail hemolymph

Science.gov (United States)

Veeramani, Vediyappan; Madhu, Rajesh; Chen, Shen-Ming; Lou, Bih-Show; Palanisamy, Jayabal; Vasantha, Vairathevar Sivasamy

2015-05-01

The biomass-derived activated carbons (ACs) have been prepared with high surface areas up to 793 m2 g-1 is by ZnCl2 activation at three different temperatures, viz. AC700, AC800, and AC900. The AC samples were characterized by a variety of analytical and spectroscopy techniques. The as-synthesized ACs were adopted for the simultaneous electrochemical detection of ascorbic acid (AA), dopamine (DA), and uric acid (UA). For comparison, reduced graphene oxide (RGO) was employed for the proposed sensor. The high surface area, modulated pore size and the presence of oxygen surface functional groups like heteroatoms (83.427% C, 1.085% N, 0.383% S, and 0.861% H) in the biomass-derived AC is found to be responsible for the excellent catalytic activities of biomolecules. Fascinatingly, the facile sensor further used to detect biomolecules levels in the snail hemolymph and human blood serum. Notably, the obtained analytical parameters for the biomolecules detection over the AC modified GCE, outperforming several carbon-based modified electrodes in literatures.

Differential Protein Expression in the Hemolymph of Bithynia siamensis goniomphalos Infected with Opisthorchis viverrini.

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Kulwadee Suwannatrai

2016-11-01

Full Text Available Bithynia siamensis goniomphalos is a freshwater snail that serves as the first intermediate host of the human liver fluke Opisthorchis viverrini. This parasite is a major public health problem in different countries throughout the Greater Mekong sub-region (Thailand, southern Vietnam, Lao PDR and Cambodia. Chronic O. viverrini infection also results in a gradual increase of fibrotic tissues in the biliary tract that are associated with hepatobiliary diseases and contribute to cholangiocarcinoma (a fatal type of bile duct cancer. Infectivity of the parasite in the snail host is strongly correlated with destruction of helminths by the snail's innate immune system, composed of cellular (hemocyte and humoral (plasma defense factors. To better understand this important host-parasite interface we applied sequential window acquisition of all theoretical spectra mass spectrometry (SWATH-MS to identify and quantify the proteins from the hemolymph of B. siamensis goniomphalos experimentally infected with O. viverrini and compare them to non-infected snails (control group. A total of 362 and 242 proteins were identified in the hemocytes and plasma, respectively. Of these, 145 and 117 proteins exhibited significant differences in expression upon fluke infection in hemocytes and plasma, respectively. Among the proteins with significantly different expression patterns, we found proteins related to immune response (up-regulated in both hemocyte and plasma of infected snails and proteins belonging to the structural and motor group (mostly down-regulated in hemocytes but up-regulated in plasma of infected snails. The proteins identified and quantified in this work will provide important information for the understanding of the factors involved in snail defense against O. viverrini and might facilitate the development of new strategies to control O. viverrini infection in endemic areas.

Proteomic characterization of the hemolymph of Octopus vulgaris infected by the protozoan parasite Aggregata octopiana.

Science.gov (United States)

Castellanos-Martínez, Sheila; Diz, Angel P; Álvarez-Chaver, Paula; Gestal, Camino

2014-06-13

The immune system of cephalopods is poorly known to date. The lack of genomic information makes difficult to understand vital processes like immune defense mechanisms and their interaction with pathogens at molecular level. The common octopus Octopus vulgaris has a high economic relevance and potential for aquaculture. However, disease outbreaks provoke serious reductions in production with potentially severe economic losses. In this study, a proteomic approach is used to analyze the immune response of O. vulgaris against the coccidia Aggregata octopiana, a gastrointestinal parasite which impairs the cephalopod nutritional status. The hemocytes and plasma proteomes were compared by 2-DE between sick and healthy octopus. The identities of 12 differentially expressed spots and other 27 spots without significant alteration from hemocytes, and 5 spots from plasma, were determined by mass spectrometry analysis aided by a six reading-frame translation of an octopus hemocyte RNA-seq database and also public databases. Principal component analysis pointed to 7 proteins from hemocytes as the major contributors to the overall difference between levels of infection and so could be considered as potential biomarkers. Particularly, filamin, fascin and peroxiredoxin are highlighted because of their implication in octopus immune defense activity. From the octopus plasma, hemocyanin was identified. This work represents a first step forward in order to characterize the protein profile of O. vulgaris hemolymph, providing important information for subsequent studies of the octopus immune system at molecular level and also to the understanding of the basis of octopus tolerance-resistance to A. octopiana. The immune system of cephalopods is poorly known to date. The lack of genomic information makes difficult to understand vital processes like immune defense mechanisms and their interaction with pathogens at molecular level. The study herein presented is focused to the comprehension of

PRESERVANDO CECROPIAIS NATIVOS EM PAISAGENS AMAZÔNICAS: UMA ESTRATÉGIA ECOLÓGICA EM SOLOS DEFICIENTES POR FÓSFORO

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Patricia Chaves de Oliveira

2010-06-01

Full Text Available A conservação de espécies prestadoras de serviços ecológicos em paisagens amazônicas como os agros-ecossistemas locais do Baixo Amazonas é de extrema importância no input de minerais através da serrapilheira produzida por estas espécies. Os cecropiais nativos, formados pela espécie Cecropia palmata, dentre outras, a qual é acumuladora de fósforo (P em tecidos foliares e cascas podem prestar um serviço ecológico em paisagens amazônicas com deficiência por fósforo. Os objetivos desta pesquisa foram a observação da translocação do fósforo em folhas ontogeneticamentes diferentes de Cecropia palmata; bem como, a análise da dinâmica do fósforo em solos sob conglomerados de Cecropia palmata. Os teores de fósforo (P em tecidos foliares de Cecropia palmata demonstraram valores elevados, em torno de 8.4 mg/kg em folhas novas e 5,6mg/kg em folhas com maturação fisiológica, sugerindo serrapilheira de alta qualidade para o ambiente. Além disso, os solos sob influência da rizosfera desta espécie apresentaram importantes hot spots de fósforo.

Pollen analysis of honey and pollen collected by Apis mellifera linnaeus, 1758 (Hymenoptera, Apidae), in a mixed environment of Eucalyptus plantation and native cerrado in Southeastern Brazil.

Science.gov (United States)

Simeão, C M G; Silveira, F A; Sampaio, I B M; Bastos, E M A F

2015-11-01

Eucalyptus plantations are frequently used for the establishment of bee yards. This study was carried on at Fazenda Brejão, northwestern region of the State of Minas Gerais, Brazil. This farm is covered both with native Cerrado vegetation (Brazilian savanna) and eucalyptus plantations. This paper reports on the botanic origin of pollen pellets and honey collected from honeybee (Apis mellifera) hives along a thirteen-month period (January 2004 to January 2005). The most frequent pollen types found in the pollen pellets during the rainy season were Trema micrantha (Ulmaceae), Copaifera langsdorffii (Fabaceae), an unidentified Poaceae, unidentified Asteraceae-2, Cecropia sp. 1 (Cecropiaceae) and Eucalyptus spp. (Myrtaceae); during the dry season the most frequent pollen types were Acosmium dasycarpum (Fabaceae), Cecropia sp. 1 (Cecropiaceae) and Eucalyptus spp. (Myrtaceae). Pollen grains of Baccharis sp. (Asteraceae), Cecropia sp. 1 (Cecropiaceae), Copaifera langsdorffii (Fabaceae), Mimosa nuda (Fabaceae), Eucalyptus spp. (Myrtaceae) and Trema micrantha (Ulmaceae) were present in the honey samples throughout the study period.

Pollen analysis of honey and pollen collected by Apis mellifera linnaeus, 1758 (Hymenoptera, Apidae, in a mixed environment of Eucalyptus plantation and native cerrado in Southeastern Brazil

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C. M. G. Simeão

Full Text Available Abstract Eucalyptus plantations are frequently used for the establishment of bee yards. This study was carried on at Fazenda Brejão, northwestern region of the State of Minas Gerais, Brazil. This farm is covered both with native Cerrado vegetation (Brazilian savanna and eucalyptus plantations. This paper reports on the botanic origin of pollen pellets and honey collected from honeybee (Apis mellifera hives along a thirteen-month period (January 2004 to January 2005. The most frequent pollen types found in the pollen pellets during the rainy season were Trema micrantha (Ulmaceae, Copaifera langsdorffii (Fabaceae, an unidentified Poaceae, unidentified Asteraceae-2, Cecropia sp. 1 (Cecropiaceae and Eucalyptus spp. (Myrtaceae; during the dry season the most frequent pollen types were Acosmium dasycarpum (Fabaceae, Cecropia sp. 1 (Cecropiaceae and Eucalyptus spp. (Myrtaceae. Pollen grains of Baccharis sp. (Asteraceae, Cecropia sp. 1 (Cecropiaceae, Copaifera langsdorffii (Fabaceae, Mimosa nuda (Fabaceae, Eucalyptus spp. (Myrtaceae and Trema micrantha (Ulmaceae were present in the honey samples throughout the study period.

A New Species of Neotropical Carpenter Ant in the Genus Camponotus (Hymenoptera: Formicidae, Apparently without Major Workers

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William Mackay

2012-01-01

Full Text Available We describe a new species of carpenter ants from Ecuador, which apparently has an obligatory relationship with the ant plants Cecropia membranacea Trécul, C. herthae Diels and C. marginalis Cuatrec. The workers are relatively small and hairy, and based on a number of collections, it does not appear to have major workers. We compare the new species to Camponotus balzani, to which it appears to be similar and which has normal major workers, and also lives in Cecropia spp.

Avaliação do efeito mutagênico do extrato hidroalcoólico bruto, por meio de bioensaios in vivo e prospecção fitoquímica de Cecropia glaziovii Sneth (embaúba, Cecropiaceae

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Victor S. Stange

Full Text Available O gênero Cecropia é conhecido popularmente por "embaúba" e apresenta várias espécies medicinais, sendo que estudos com a espécie C. glaziovii Sneth indicam que o extrato aquoso apresenta efeitos broncodilatador, anti-hipertensivo e antidepressivo, provavelmente, atribuídos às catequinas, procianidinas e flavonóides. O presente estudo objetivou avaliar os potenciais efeitos tóxico, citotóxicos, clastogênicos e aneugênicos do extrato bruto hidroalcoólico de folhas de C. glaziovii, por meio da dose letal média (DL50, da análise de micronúcleo em células de medula óssea de roedores e do teste com Allium cepa e realizar a prospecção fitoquímica. Os resultados demonstraram que as concentrações testadas não causaram toxicidade aguda, nem apresentaram atividade clastogênica e aneugênica, sugerindo que o extrato de C. glaziovii não interferem na divisão celular. A prospecção fitoquímica indicou a presença de taninos, flavonóides, fenóis, antraquinonas, cumarinas, catequinas, proteínas, açúcares redutores, depsídeos/depsidonas e triterpenos. Mesmo assim, esses resultados não prescrevem o consumo terapêutico da espécie, sem continuidade da sua avaliação e do cumprimento de todas as etapas pré-clínicas e clínicas.

Production of polyclonal antibodies for lectin from Anticarsia gemmatalis hemolymph Produção de anticorpos policlonais para lectina de hemolinfa de Anticarsia gemmatalis

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Mario Augusto Ono

2005-10-01

Full Text Available The velvet bean caterpillar Anticarsia gemmatalis promotes extensive damage to soybean and is controlled frequently by chemical insecticides. Due to risks to human and animal health as well as the environment, new approaches were developed to A. gemmatalis control such as the bioinsectide Baculoviru anticarsia. The development of resistance in A. gemmatalis populations treated along several generations with B. anticarsia was reported under laboratory conditions. The insects show complex mechanisms against microorganism infection, such as the lectins, that work as recognition molecules. The aim of this work was to develop polyclonal antibodies to A. gemmatalis lectin. The lectin activity in A. gemmatalis caterpillar hemolymph was evaluated using erythrocytes from human, rabbit, mouse, sheep and cow. Only bovine erythrocytes were not agglutinated by lectin. The rabbit erythrocytes showed stronger reactivity (1:512. Therefore the polyclonal antibodies were raised in rabbit immunized with autologous erythrocytes sensitized with lectin. The antibody to lectin showed a titer of 1:8 in immunodiffusion test. In this study we described a simple method for antibody production against hemolymph lectin without expensive purification techniques. A lagarta de Anticarsia gemmatalis promove extensos danos na cultura da soja e seu controle é geralmente baseado na aplicação de inseticidas químicos. Devido aos riscos à saúde humana, animal e ao meio ambiente, métodos alternativos de controle tem sido desenvolvidos como o bioinseticida Baculovirus anticarsia. Há relatos de desenvolvimento de resistência em populações de A. gemmatalis submetidas, em laboratório, ao tratamento com baculovirus durante várias gerações. Os insetos apresentam mecanismos elaborados de proteção contra agentes infecciosos, como as lectinas, que atuam como moléculas de reconhecimento. Assim, o objetivo deste estudo foi desenvolver anticorpos policlonais para lectina de A

Florística e estrutura de fragmento florestal em área de transição na Amazônia Matogrossense no município de Sinop Floristics and structure of a forest fragment at a transitional zone at the Amazon in Mato Grosso State, Municipality of Sinop

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Rosalia de Aguiar Araujo

2009-01-01

Full Text Available A fragmentação do ambiente é intensa na amazônia matogrossense e é esperado como consequência, além do desaparecimento da vegetação original, que a flora dos fragmentos seja transicional, com elementos de floresta amazônica e de cerrado e que já apresentem elevada presença de famílias e espécies pioneiras. Este trabalho objetivou avaliar a composição florística e obter parâmetros fitossociológicos de componentes arbóreos presentes em um fragmento urbano no município de Sinop, Mato Grosso com vistas a confirmar sua classificação como vegetação de transição e detectar indícios de perturbação pela fragmentação. A vegetação foi amostrada pela instalação de 25 parcelas permanentes de 20 x 20m, onde foram amostrados os indivíduos com CAP igual ou superior a 15 cm à altura de 1,30m do solo. A densidade total da área foi de 1555 ind./ha, distribuídos em 37 famílias botânicas, 81 gêneros e 113 espécies. A família mais representativa foi Leguminosae com 14 espécies. As espécies que mais contribuíram em abundância e apresentaram maior frequência foram: Cecropia sciadophylla, Cecropia sp., Bellucia grossularioides e Vismia guianensis. O índice de Shannon (H' foi de 3,55, considerado alto para uma floresta de transição. A equabilidade de Pielou foi de 0,75, sugerindo grande dominância de poucas espécies. As espécies que mais se destacaram em ordem decrescente de VI (% foram Cecropia sp., Bellucia grossularioides, Qualea ingens, Cecropia sciadophylla, Vismia guianensis, Miconia prasina, Trattinickia burserifolia, Unonopsis guatterioides e Schefflera vinosa. O remanescente apresenta uma flora mista amazônica e de cerrado, confirmando ser ecótono e a abundância de pioneiras sugere distúrbios. Esta floresta protege espécies madeireiras e frutíferas com grande potencial para uso múltiplo, podendo ser um espaço educativo com vistas a conservação e manejo sustentável.Fragmentation is intense in

Moraceae

NARCIS (Netherlands)

Berg, C.C.

1980-01-01

Trees or shrubs, in Ficus often epiphytic and strangling, in Dorstenia herbs, laticiferous except for Cecropia. Leaves alternate, lamina entire or lobed, venation pinnate, palmate or radiate, stipules often amplexicaul. Plants monoecious or dioecious, flowers unisexual, close together in bisexual or

Plantas de ocorrência espontânea como substratos alternativos para fitoseídeos (Acari, Phytoseiidae em cultivos de seringueira Hevea brasiliensis Muell. Arg. (Euphorbiaceae Weeds as alternative substrates to phytoseiids (Acari, Phytoseiidae in rubber tree Hevea brasilienis, Muell. Arg. (Euphorbiaceae cultivation

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Marcos Roberto Bellini

2005-03-01

Full Text Available O objetivo deste estudo foi determinar as espécies de plantas espontâneas em áreas de cultivo de seringueira que poderiam servir com reservatório de ácaros predadores. O trabalho foi conduzido em Olímpia, Estado de São Paulo, Brasil, em dois cultivos de seringueira. A cada três meses, cerca de 1.000 cm³ de folhas de cada uma das cinco espécies de plantas espontâneas dominantes em cada cultivo foi tomada para determinar os fitoseídeos. Como as plantas espontâneas dominantes variaram durante o estudo, um total de 20 espécies foi verificado. Somente Cecropia sp. foi dominante em todo estudo, em ambos os cultivos. Um total de 336 fitoseídeos pertencentes às seguintes espécies foi encontrado: Euseius citrifolius Denmark & Muma, 1970, E. concordis (Chant, 1959, Galendromus annectens (DeLeon, 1958, Iphiseiodes zuluagai Denmark & Muma, 1972, Neoseiulus anonymus (Chant & Baker, 1965 e N. tunus (DeLeon, 1967. E. citrifolius (189 espécimes e N. tunus (138 espécimes foram os mais abundantes. A maior abundância (231espécimes e diversidade (5 espécies foram observadas sobre Cecropia sp. Sobre esta planta foi encontrado o maior número de ácaros por amostra (29 espécimes, seguida por Piper duncum Linnaeus (22, Guarea sp. (18 e Ageratum conyzoides Linnaeus (12. E. citrifolius tem sido determinado como um dos ácaros predadores mais abundantes sobre seringueiras cultivadas na região onde este trabalho foi conduzido. Cecropia sp. parece ser uma das plantas espontâneas mais importantes consideradas como reservatório de E. citrifolius porque está continuamente presente nos cultivos. Estudos complementares poderiam indicar a viabilidade de se manejar esta planta nas áreas de cultivo de seringueira para permitir o controle biológico dos ácaros-praga desta cultura.The objective of this study was to determine weed species of rubber tree cultivation areas that could serve as reservoirs of predatory mites. The work was conducted in Ol

Vitellogenin RNAi halts ovarian growth and diverts reproductive proteins and lipids in young grasshoppers.

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Tokar, Derek R; Veleta, Katherine A; Canzano, Joseph; Hahn, Daniel A; Hatle, John D

2014-11-01

Reduced reproduction extends lifespan of females in many animals. To test the effects of reproduction on storage of macronutrients, we block reproductive output in the lubber grasshopper by injecting RNAi against the precursor to egg-yolk protein, vitellogenin, in early adulthood. Controls were injected with either buffer or RNAi against the major storage protein in the hemolymph, hexamerin-90. Vitellogenin RNAi greatly reduced both levels of mRNA for vitellogenin and ovarian growth, in comparison to both controls. Fat body mass was increased upon vitellogenin RNAi, but concentrations of the three hexameric storage proteins from the hemolymph were not. Surprisingly, hemolymph vitellogenin levels were increased upon vitellogenin RNAi. Total reproductive protein (hemolymph vitellogenin plus ovarian vitellin) was unchanged by vitellogenin RNAi, as reproductive protein was diverted to the hemolymph. Similarly, the increased lipid storage upon vitellogenin RNAi was largely attributable to the reduction in lipid in the ovary, due to decreased ovarian growth. A BLAST search revealed that the 515 bp sequence of vitellogenin used for RNAi had three 11 bp regions identical to the vitellogenin receptor of the cockroach Leucophaea maderae. This suggests that our treatment, in addition to reducing levels of vitellogenin transcript, may have also blocked transport of vitellogenin from the hemolymph to the ovary. This would be consistent with halted ovarian growth simultaneous with high levels of vitellogenin in the hemolymph. Nonetheless, the accumulation of vitellogenin, instead of hexameric storage proteins, is inconsistent with a simple model of the trade-off between reproduction and storage. This was observed in young females; future studies will address whether investment of proteins may shift to the soma as individuals age. Overall, our results suggest that blockage of reproduction in young grasshoppers redirects lipids to storage and reproductive proteins to the hemolymph

Learning from our mistakes: minimizing problems with invasive biofuel plants

CSIR Research Space (South Africa)

Richardson, DM

2011-03-01

Full Text Available -legume service trees (e.g., Cecropia spp.); 168  Fast-growing timber trees (e.g., Eucalyptus spp., Casuarina spp.); 169  High-value timber trees (e.g., Cedrela odorata, Cordia alliodora); 170  Fruit trees (e.g., Citrus spp., Psidium guajava). 171 172...

Triagem fitoquímica e avaliação das atividades trombolítica e citotóxica de Cecropia hololeuca Miq. (Urticaceae, Lippia alba (Mill. N.E.Br. ex P. Wilson (Verbenaceae e Zanthoxylum rhoifolium Lam (Rutaceae

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João Victor Dutra GOMES1 ; Rafael Destefani FAITANIN

2016-03-01

Full Text Available Cecropia hololeuca, Lippia alba, e Zanthoxylum rhoifolium são espécies vegetais de ocorrência natural no Brasil, características de regiões de Mata Atlântica e, dentro de suas finalidades medicinais, estão incluídas ação anti-hipertensiva, digestiva e no tratamento de malária. Neste estudo, foram avaliados o perfil fitoquímico e as atividades citotóxica e trombolítica do extrato etanólico das folhas destas espécies vegetais. O perfil químico dos extratos foi obtido por meio de ensaios fitoquímicos clássicos, a fim de identificar as classes químicas presentes. A toxicidade preliminar foi avaliada frente Artemia salina e a atividade trombolítica foi determinada in vitro a partir da lise de coágulo de sangue humano. Os testes fitoquímicos indicaram a presença de alcaloides e flavonoides nos três extratos investigados. Saponinas, triterpenos e naftoquinonas foram detectados apenas em C. hololeuca. Essa não demonstrou toxicidade frente Artemia salina (DL50 >1000 ppm, enquanto Z. rhoifolium apresentou DL50 igual a 719,44 ppm e L. alba <250 ppm, o que indica atenção quanto à segurança no uso desta planta medicinal. A atividade trombolítica de L. alba e C. hololeuca foi de 6,43 ±2,08 e 9,64 ±1,83 %, respectivamente, mostrando baixa atividade. Por sua vez, Z. rhoifolium alcançou lise de 24,71 ±10,52 %, indicando promissora atividade (p <0,001. A atividade trombolítica de Z. rhoifolium justifica novos estudos, a fim de investigar os componentes responsáveis pela atividade. Perante a literatura, este é o primeiro relato da avaliação da atividade trombolítica destas espécies vegetais. Os resultados encontrados neste trabalho contribuem para o conhecimento químico-biológico das respectivas espécies.

Effects of prolonged acclimation to cold on the extra--and intracellular acid-base status in the land snail Helix lucorum (L.).

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Staikou, A; Stiakakis, M; Michaelidis, B

2001-01-01

The aim of this study was to examine the effect of prolonged acclimation to cold on the acid-base status of extra- and intracellular fluids in the land snail Helix lucorum. For this purpose, acid-base parameters in the hemolymph and tissues were determined. In addition, the buffer values of hemolymph and tissues were determined in order to examine whether they change in the snails during acclimation to cold. According to the results presented, there is an inverse pH-temperature relationship in the hemolymph within the first day of acclimation, which is consistent with alphastat regulation. The Pco2 decreased, and pH in the hemolymph (pH(e)) increased by 0.32 U within the first day of acclimation to cold, which corresponds to a change of 0.013 U degrees C(-1). After the first day of acclimation, Pco2 increased in the hemolymph, resulting in a significant drop in pH(e) by 90 d of acclimation to cold. Acclimation of snails to low temperatures did not change the buffer value of the hemolymph. Also, intracellular pH (pH(i)) and intracellular buffer values remained stable during acclimation to cold for prolonged periods. The latter results in conjunction with those obtained by the in vitro determination of the passive component of intracellular fluids indicate an active regulation of pH(i) in H. lucorum during acclimation to cold.

Assessment of electron beam irradiation induced proteomic changes and its effect on the development of silkworm, Bombyx mori (Bombycidae: Lepidoptera

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Mani Kannan

2016-01-01

Full Text Available The present study has been designed to determine the hemolymph protein changes induced by EBI using Microtron (ranges from 20 to 100 Gy on the 5th instar larvae of silkworm, Bombyx mori using SDS–PAGE. The EBI did not caused any impact on 5th instar larval hemolymph proteins, however a significant reduction of pupal hemolymph proteins such as lipophorin (250 kDa, vitellogenin (180 kDa, storage protein (76–80 kDa and a 30 kDa protein was observed through SDS–PAGE and densitometry analysis. These proteins are known to play a crucial role in various developmental processes including transport of lipids, as amino acid reservoir for providing precursors for egg and cuticle formation and immunity of B. mori. Further, a decrease of antioxidant enzymes such as Superoxide Dismutase (SOD and Catalase (CAT in the EBI treated larval and pupal hemolymph was observed. In addition, a negative influence on growth characteristics and appearance of pupal deformity was noted. This may be due to damage in hemolymph proteins, when the larvae were exposed to EBI at >80 Gy.

Abundância e frugivoria da quiropterofauna (Mammalia, chiroptera de um fragmento no noroeste do Estado do Paraná, Brasil = Chiropterofauna abundance and frugivory in a forest remnant in northwestern Paraná State, Brazil

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João Eduardo Cavalcanti Brito

2010-07-01

Full Text Available A abundância e a frugivoria de morcegos que compõem a taxocenose em uma área de mata ripária, à margem esquerda do rio Ivaí, foram foco do presente estudo. O Recanto Marista possui 57,6 hectares, dos quais 40,8 são cobertos por Floresta Estacional Semidecidual, situado no município de Doutor Camargo, região Noroeste do Estado do Paraná. Foram realizadas 14 noites de capturas de morcegos de maio de 2007 a janeiro de 2008, com redesneblina (7 x 2,5 m, totalizando 13.475 m² h de esforço amostral, distribuído em 72h de esforço. Foram capturados 193 indivíduos, representantes de dez espécies, pertencentes a duas famílias: Phyllostomidae (Artibeus lituratus, Sturnira lilium, Carollia perspicillata, Artibeus cf. fimbriatus, Artibeus planirotris, Desmodus rotundus e Pygoderma bilabiatum e Vespertilionidae (Myotis nigricans, Eptesicus sp. e Lasiurus blossevillii. Um representante da família Molossidae (Molossus rufus foi encontrado morto no solo. Foram consumidos frutos pertencentes às famílias Moraceae (Ficus guaranitica, Ficus insipida, Ficus sp. e Maclura tinctoria, Solanaceae (Solanum aspero-lanatum e Solanum sp., Piperaceae (Piper aduncum, Piper amalago e Piper sp. e Urticaceae (Cecropia pachystachya e Cecropia sp..This study aims to evaluate the abundance and frugivory of bats from the Recanto Marista, a small riparian forest remnant in the margins of the Ivaí river. The Recanto Marista has 57.6 ha, of which 40.8 ha are covered by semideciduous seasonal forest and is located in the Doutor Camargo municipality. Collections were conducted from May 2007to January 2008 using mist nets (7 x 2.5 m totaling 13,475 m² h and comprising about 72 hours. Ten species were found pertaining to two families, Phyllostomidae (Artibeus lituratus, Sturnira lilium, Carollia perspicillata, Artibeus cf. fimbriatus, Artibeus planirotris, Desmodus rotundus and Pygoderma bilabiatum and Vespertilionidae (Myotis nigricans, Eptesicus sp. and Lasiurus

Abundância e frugivoria da quiropterofauna (Mammalia, chiroptera de um fragmento no noroeste do Estado do Paraná, Brasil - doi: 10.4025/actascibiolsci.v32i3.5351 Chiropterofauna abundance and frugivory in a forest remnant in northwestern Paraná State, Brazil - doi: 10.4025/actascibiolsci.v32i3.5351

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Janaina Gazarini

2010-09-01

Full Text Available A abundância e a frugivoria de morcegos que compõem a taxocenose em uma área de mata ripária, à margem esquerda do rio Ivaí, foram foco do presente estudo. O Recanto Marista possui 57,6 hectares, dos quais 40,8 são cobertos por Floresta Estacional Semidecidual, situado no município de Doutor Camargo, região Noroeste do Estado do Paraná. Foram realizadas 14 noites de capturas de morcegos de maio de 2007 a janeiro de 2008, com redes-neblina (7 x 2,5 m, totalizando 13.475 m² h de esforço amostral, distribuído em 72h de esforço. Foram capturados 193 indivíduos, representantes de dez espécies, pertencentes a duas famílias: Phyllostomidae (Artibeus lituratus, Sturnira lilium, Carollia perspicillata, Artibeus cf. fimbriatus, Artibeus planirotris, Desmodus rotundus e Pygoderma bilabiatum e Vespertilionidae (Myotis nigricans, Eptesicus sp. e Lasiurus blossevillii. Um representante da família Molossidae (Molossus rufus foi encontrado morto no solo. Foram consumidos frutos pertencentes às famílias Moraceae (Ficus guaranitica, Ficus insipida, Ficus sp. e Maclura tinctoria, Solanaceae (Solanum aspero-lanatum e Solanum sp. , Piperaceae (Piper aduncum, Piper amalago e Piper sp. e Urticaceae (Cecropia pachystachya e Cecropia sp..This study aims to evaluate the abundance and frugivory of bats from the Recanto Marista, a small riparian forest remnant in the margins of the Ivaí river. The Recanto Marista has 57.6 ha, of which 40.8 ha are covered by semideciduous seasonal forest and is located in the Doutor Camargo municipality. Collections were conducted from May 2007 to January 2008 using mist nets (7 x 2.5 m totaling 13,475 m² h and comprising about 72 hours. Ten species were found pertaining to two families, Phyllostomidae (Artibeus lituratus, Sturnira lilium, Carollia perspicillata, Artibeus cf. fimbriatus, Artibeus planirotris, Desmodus rotundus and Pygoderma bilabiatum and Vespertilionidae (Myotis nigricans, Eptesicus sp. and Lasiurus

A novel insect defensin mediates the inducible antibacterial activity in larvae of the dragonfly Aeschna cyanea (Paleoptera, Odonata)

NARCIS (Netherlands)

Bulet, P.; Cociancich, S.; Reuland, M.; Sauber, F.; Bischoff, Rainer; Hegy, G.; Van Dorsselaer, A.; Hetru, C.; Hoffmann, J.A.

1992-01-01

The injection of low doses of bacteria into the aquatic larvae of dragonflies (Aeschna cyanea, Odonata, Paleoptera) induces the appearance in their hemolymph of a potent antibacterial activity. We have isolated a 38-residue peptide from this hemolymph which is strongly active against Gram-positive

Unveiling the oxidative metabolism of Achatina fulica (Mollusca: Gastropoda) experimentally infected to Angiostrongylus cantonensis (Nematoda: Metastrongylidae).

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Tunholi-Alves, Vinícius Menezes; Tunholi, Victor Menezes; Garcia, Juberlan; Mota, Esther Maria; Castro, Rosane Nora; Pontes, Emerson Guedes; Pinheiro, Jairo

2018-06-01

For the first time, alterations in the oxidative metabolism of Achatina fulica experimentally infected with different parasite loads of Angiostrongylus cantonensis were determined. For this, the hemolymph activities of lactate dehydrogenase (LDH) and hexokinase and the glucose concentrations in the hemolymph, as well as the polysaccharide reserves in the digestive gland and cephalopedal mass, were assessed. Additionally, the contents of some carboxylic acids in the hemolymph of infected and uninfected snails were determined by high-performance liquid chromatography (HPLC), permitting a better understanding of the alterations related to the host's oxidative metabolism. As the main results, activation of oxidative pathways, such as the glycolytic pathway, was demonstrated in response to the increase in the activity of hexokinase. This tendency was confirmed by the decrease in the contents of glucose in the hemolymph of parasitized snails, indicating that the infection by A. cantonensis alters the host's metabolism, and that these changes are strongly influenced by the parasite load. This metabolic scenario was accompanied by activation of the anaerobic fermentative metabolism, indicated not only by an increase in the activity of (LDH), but also by a reduction of the content of pyruvic acid and accumulation of lactic acid in the hemolymph of parasitized snails. In this circumstance, maintenance of the host's redox balance occurs through activation of the fermentative pathways, and LDH plays a central role in this process. Together, the results indicate that A. cantonensis infection induces activation of the anaerobic metabolism of A. fulica, characterized not only by the accumulation of lactic acid, but also by a reduction in the pyruvic acid and oxalic acid contents in the hemolymph of the infected snails.

Caterpillars and fungal pathogens: two co-occurring parasites of an ant-plant mutualism.

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Olivier Roux

Full Text Available In mutualisms, each interacting species obtains resources from its partner that it would obtain less efficiently if alone, and so derives a net fitness benefit. In exchange for shelter (domatia and food, mutualistic plant-ants protect their host myrmecophytes from herbivores, encroaching vines and fungal pathogens. Although selective filters enable myrmecophytes to host those ant species most favorable to their fitness, some insects can by-pass these filters, exploiting the rewards supplied whilst providing nothing in return. This is the case in French Guiana for Cecropia obtusa (Cecropiaceae as Pseudocabima guianalis caterpillars (Lepidoptera, Pyralidae can colonize saplings before the installation of their mutualistic Azteca ants. The caterpillars shelter in the domatia and feed on food bodies (FBs whose production increases as a result. They delay colonization by ants by weaving a silk shield above the youngest trichilium, where the FBs are produced, blocking access to them. This probable temporal priority effect also allows female moths to lay new eggs on trees that already shelter caterpillars, and so to occupy the niche longer and exploit Cecropia resources before colonization by ants. However, once incipient ant colonies are able to develop, they prevent further colonization by the caterpillars. Although no higher herbivory rates were noted, these caterpillars are ineffective in protecting their host trees from a pathogenic fungus, Fusarium moniliforme (Deuteromycetes, that develops on the trichilium in the absence of mutualistic ants. Therefore, the Cecropia treelets can be parasitized by two often overlooked species: the caterpillars that shelter in the domatia and feed on FBs, delaying colonization by mutualistic ants, and the fungal pathogen that develops on old trichilia. The cost of greater FB production plus the presence of the pathogenic fungus likely affect tree growth.

A new record of fungus-beetle symbiosis in Scolytodes bark beetles (Scolytinae, Curculionidae, Coleoptera)

Czech Academy of Sciences Publication Activity Database

Hulcr, J.; Kolařík, Miroslav; Kirkendall, L.

2007-01-01

Roč. 43, - (2007), s. 151-159 ISSN 0334-5114 Grant - others:GA ČR(CZ) GD206/03/H034 Program:GD Institutional research plan: CEZ:AV0Z50200510 Source of funding: V - iné verejné zdroje Keywords : scolytodes unipunctatus * cecropia * raffaela Subject RIV: EE - Microbiology, Virology Impact factor: 0.598, year: 2007

It takes time to see the menu from the body: an experiment on stable isotope composition in freshwater crayfishes

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Jussila J.

2015-01-01

Full Text Available For many applications and ecological studies in which wild individuals are brought to laboratory it would be essential to know accurately how fast novel diet is reflected in composition of different tissues. To study the effects of two different diets on the stable isotope composition of freshwater crayfish muscle and hemolymph, we conducted a three month experiment on noble crayfish (Astacus astacus and signal crayfish (Pacifastacus leniusculus by feeding them sweet corn (Zea mays or Baltic herring (Clupea harengus membras as novel food. During the experiment, the crayfish were given 0.4 g of selected food daily and the amount consumed was recorded. The samples for the stable isotope analyses were taken at the commencement of the experiment (initial control and three times (hemolymph or twice (muscle tissue during the experiment. We found that stable isotope changes can be similarly, and rather slowly, detected from muscle tissue and hemolymph under studied conditions. Hemolymph sampling, being non-lethal, can be recommended as a practical sampling method. Our results confirm earlier reports according to which diet changes reflect to crayfish isotope ratios slowly implying that isotope ratios indicate long-term diet.

Estudo quantitativo de metais presentes na hemolinfa de Biomphalaria glabrata (Gastropoda, infectadas e não infectadas com Schistosoma mansoni Quantitative study of metal present in the hemolymph of Biomphalaria glabrata (Gastropoda, infected and uninfected with Schistosoma mansoni

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Marco Antonio Vasconcelos Santos

2005-04-01

Full Text Available Inicialmente, desenvolveu-se um estudo para quantificar e comparar as concentrações de alguns metais presentes em duas amostras de hemolinfa do caramujo Biomphalaria glabrata (infectados e não-infectados com Schistosoma mansoni. A espectrometria de emissão óptica com fonte de plasma induzido (ICP-OES, foi utilizada para analisar os metais nas duas amostras. Os metais estudados foram: alumínio, cálcio, cádmio, cobalto, cromo, cobre, ferro, potássio, magnésio, manganês, chumbo e zinco. Os resultados mostram que, a princípio, os metais não são fatores determinantes no processo de defesa desses organismos contra este parasita, quando presente nos seus tecidos.We conducted a preliminary study to quantify and compare two concentrations of the same metals present in the hemolymph of snail Biomphalaria glabrata. In this context, we used Induction Coupled Plasma Optical Emission Spectroscopy technique (ICP-OES, to analyze the metals in the two samples (snails infected and not infected with Schistosoma mansoni. The metals studied were: aluminum, calcium, cadmium, cobalt, chromium, copper, iron, potassium, magnesium, manganese, lead and zinc. Preliminary results showed that such metals are not involved in the defense of these organisms against the parasite, when present in their tissues.

A feeding protocol for delivery of agents to assess development in Varroa mites.

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Ana R Cabrera

Full Text Available A novel feeding protocol for delivery of bio-active agents to Varroa mites was developed by providing mites with honey bee larva hemolymph supplemented with cultured insect cells and selected materials delivered on a fibrous cotton substrate. Mites were starved, fed on treated hemolymph to deliver selected agents and then returned to bee larvae. Transcript levels of two reference genes, actin and glyceraldehyde 3-phosphate dehydrogenase (GAPDH, as well as for nine selected genes involved in reproductive processes showed that the starvation and feeding protocol periods did not pose a high level of stress to the mites as transcript levels remained comparable between phoretic mites and those completing the protocol. The feeding protocol was used to deliver molecules such as hormone analogs or plasmids. Mites fed with Tebufenozide, an ecdysone analog, had higher transcript levels of shade than untreated or solvent treated mites. In order to extend this feeding protocol, cultured insect cells were incorporated to a final ratio of 1 part cells and 2 parts hemolymph. Although supplementation with Bombyx mori Bm5 cells increased the amount of hemolymph consumed per mite, there was a significant decrease in the percentage of mites that fed and survived. On the other hand, Drosophila melanogaster S2 cells reduced significantly the percentage of mites that fed and survived as well as the amount of hemolymph consumed. The feeding protocol provides a dynamic platform with which to challenge the Varroa mite to establish efficacy of control agents for this devastating honey bee pest.

Food niche overlap among neotropical frugivorous bats in Costa Rica

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Jorge E Lopez

2007-03-01

Full Text Available Food habits of 15 species of frugivorous bats were studied at La Selva Biological Station, Costa Rica. Eight hundred and fifty-four (854 fecal samples and 169 samples from fruit parts and seeds discarded by bats beneath feeding roosts were analyzed. During eight months of study, 47 fruit species consumed by bats were identified. Five plant genera (Cecropia, Ficus, Piper, Solanum, and Vismia constituted 85% of all plants found in fecal samples. Feeding niche breadth differed significantly among the six most common species of frugivorous bats (Artibeus jamaicensis, Carollia sowelli, C. castanea, C. perspicillata, Dermanura sp., and Glossophaga commissarisi. All species, except for Dermanura sp., showed a diet dominated by one or two plant species. This suggests a pattern of resource partitioning at a generic level, in which Carollia consumed mainly Piper, Artibeus consumed Ficus and Cecropia, and Glossophaga consumed Vismia. Cluster analysis revealed higher values of food niche overlap in congeneric species than among species of different genera. Results show that if food is a limiting factor, mechanisms other than trophic selection must reduce interspecific interference or competition for food in this frugivorous bat guild. Rev. Biol. Trop. 55 (1: 301-313. Epub 2007 March. 31.Estudiamos los hábitos alimentarios de 15 especies de murciélagos frugívoros en la Estación Biológica La Selva. Se analizó 854 muestras de heces y 169 muestras de restos de frutos y semillas en comederos. Durante ocho meses de estudio, se identificó 47 especies de frutos, que fueron consumidos por los murciélagos. Cinco géneros de plantas (Cecropia, Ficus, Piper, Solanum y Vismia constituyeron el 85% de los hallazgos en las muestras de heces y los comederos. La amplitud de nicho trófico difirió significativamente entre las seis especies de murciélagos frugívoros más frecuentemente capturados (Artibeus jamaicensis, Carollia sowelli, C. castanea, C. perspicillata

Larval serum proteins of the gypsy moth, Lymantria dispar: Allometric changes during development suggest several functions for arylphorin and lipophorin

International Nuclear Information System (INIS)

Karpells, S.T.

1989-01-01

Storage proteins are the major nutritive intermediates in insects and although the serum storage proteins are relatively well studied, definitive roles for many of them have yet to be established. To further characterize their roles in development and to establish quantitative baselines for future studies, two serum proteins, arylphorin (Ap) and lipophorin (Lp), of the gypsy moth, Lymantria dispar, were studied. Ap and Lp, isolated from larval hemolymph, were partially characterized biochemically and immunologically. Hemolymph concentrations throughout larval development were determined using quantitative immunoelectrophoresis and absolute hemolymph amounts of protein were determined by measuring hemolymph volume. Cyclic fluctuations in hemolymph concentrations of Ap in particular correlated with each molting cycle and an increase in Lp levels just prior to pupation suggest a metamorphic change in the role or demand for the protein. Sexual dimorphism in protein concentrations are explained in part by the sexual dimorphism in the number of larval instars. In fact, an additional instar of Ap accumulation in the female gypsy moth is suggested to compensate for the lack of a female-specific storage protein in this species. The last two days of each instar were found to be the optimum time to sample protein concentration with minimum variance. Allometric relationships among Ap accumulation, Lp accumulation and weight gain were uncovered. Ap labelled with [ 14 C]-N-ethylmaleimide was shown to be incorporated into newly synthesized cuticle and setae during a larval-larval molt. The antiserum developed against L. dispar Ap was used to identify the Ap of Trichoplusia in and study Ap titers in parasitized T. in larvae. The antiserum was also used to determine the immunological relatedness of 5 species of Lepidoptera

Cadmium transport by the gut and Malpighian tubules of Chironomus riparius

International Nuclear Information System (INIS)

Leonard, Erin M.; Pierce, Laura M.; Gillis, Patricia L.; Wood, Chris M.; O'Donnell, Michael J.

2009-01-01

Many aquatic insects are very insensitive to cadmium in short-term laboratory studies. LC50 values for larvae of the midge Chironomus riparius are over 25,000 times the Criterion Maximum Concentration in the United States Environmental Protection Agency (U.S. EPA (2000)) species sensitivity distribution (SSD). Excretion or sequestration of cadmium may contribute to insensitivity and we have therefore examined cadmium transport by isolated guts and renal tissues of C. riparius larvae. Regional differences of Cd transport along the gut were identified using a Cd 2+ -selective microelectrode in conjunction with the Scanning Ion-Selective Electrode Technique (SIET). Cd is transported into the anterior midgut (AMG) cells from the lumen and out of the cells into the hemolymph. The transport of Cd from the gut lumen to the hemolymph exposes other tissues such as the nervous system and muscles to Cd. The gut segments which remove Cd from the hemolymph at the highest rate are the posterior midgut (PMG) and the ileum. In addition, assays using an isolated Malpighian (renal) tubule preparation have shown that the Malpighian tubules (MT) both sequester and secrete Cd. For larvae bathed in 10 μmol l -1 Cd, the tubules can secrete the entire hemolymph burden of Cd in ∼15 h.

Antioxidant activity of Paraguayan plant extracts.

Science.gov (United States)

Velázquez, E; Tournier, H A; Mordujovich de Buschiazzo, P; Saavedra, G; Schinella, G R

2003-02-01

The antioxidant properties of six medical herbs used in the traditional Paraguayan medicine were studied using free radical-generating systems. The methanol extracts from Aristolochia giberti, Cecropia pachystachya, Eugenia uniflora, Piper fulvescens, Schinus weinmannifolia and Schinus terebinthifolia protected against enzymatic and non-enzymatic lipid peroxidation in microsomal membranes of rat. C. pachystachya, E. uniflora, S. weinmannifolia and S. terebinthifolia showed the highest scavenging activity on the superoxide and DPPH radicals.

Late Holocene paleoenvironments of the floodplain of the Solimões River, Central Amazonia, based on the palynological record of Lake Cabaliana

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Natália de Paula Sá

Full Text Available ABSTRACT The core PD-67 of 160 cm depth was collected from the delta of Lake Cabaliana situated on the Solimões River. Seventeen samples were removed for palynological and sedimentological analysis and three for radiocarbon analysis. Two dry periods, both in the Late Holocene, were observed (2800-2550 cal yr BP, 1450-550 cal yr BP separated by a wetter phase (2550-1450 cal yr BP. In 2800-2550 cal yr BP, varzea forests of Alchornea, Symmeria, Cecropia, Alternanthera and Asteraceae were predominant. Beginning in 2,550-1450 cal yr BP, the varzea was characterized by pioneer elements, such as Cassia, Laetia, Mabea, Symmeria and Cecropia, and by the expansion of Poaceae, Cyperaceae, Sagittaria, Montrichardia and Asteraceae. In 1450-550 cal yr BP the succession of varzea continued with Pseudobombax, Laetia, Luehea/Lueheopsis and Ryanaea increasing simultaneously with the terra firme vegetation of Rutaceae, Sapotaceae, Styrax, Scleronema, Anthurium, Araceae, pteridophytes and Pariana. The successional dynamics at Lake Cabaliana indicated that the local varzea had become established recently, and is composed of a mosaic of different successional stages of vegetation influenced mainly by flood pulse and variation in rainfall. It is therefore possible to propose that the recent climate history of Central Amazonia reflects changes in rainfall patterns in the basin.

Flight metabolism in Panstrongylus megistus (Hemiptera: Reduviidae): the role of carbohydrates and lipids

OpenAIRE

Canavoso, Lilián E; Stariolo, Raúl; Rubiolo, Edilberto R

2003-01-01

The metabolism of lipids and carbohydrates related to flight activity in Panstrongylus megistus was investigated. Insects were subjected to different times of flight under laboratory conditions and changes in total lipids, lipophorin density and carbohydrates were followed in the hemolymph. Lipids and glycogen were also assayed in fat body and flight muscle. In resting insects, hemolymph lipids averaged 3.4 mg/ml and significantly increased after 45 min of flight (8.8 mg/ml, P < 0.001). High-...

Natural history-driven, plant-mediated RNAi-based study reveals CYP6B46's role in a nicotine-mediated antipredator herbivore defense.

Science.gov (United States)

Kumar, Pavan; Pandit, Sagar S; Steppuhn, Anke; Baldwin, Ian T

2014-01-28

Manduca sexta (Ms) larvae are known to efficiently excrete ingested nicotine when feeding on their nicotine-producing native hostplant, Nicotiana attenuata. Here we describe how ingested nicotine is co-opted for larval defense by a unique mechanism. Plant-mediated RNAi was used to silence a midgut-expressed, nicotine-induced cytochrome P450 6B46 (CYP6B46) in larvae consuming transgenic N. attenuata plants producing MsCYP6B46 dsRNA. These and transgenic nicotine-deficient plants were planted into native habitats to study the phenotypes of larvae feeding on these plants and the behavior of their predators. The attack-behavior of a native wolf spider (Camptocosa parallela), a major nocturnal predator, provided the key to understanding MsCYP6B46's function: spiders clearly preferred CYP6B46-silenced larvae, just as they had preferred larvae fed nicotine-deficient plants. MsCYP6B46 redirects a small amount (0.65%) of ingested nicotine from the midgut into hemolymph, from which nicotine is exhaled through the spiracles as an antispider signal. CYP6B46-silenced larvae were more susceptible to spider-attack because they exhaled less nicotine because of lower hemolymph nicotine concentrations. CYP6B46-silenced larvae were impaired in distributing ingested nicotine from midgut to hemolymph, but not in the clearing of hemolymph nicotine or in the exhalation of nicotine from hemolymph. MsCYP6B46 could be a component of a previously hypothesized pump that converts nicotine to a short-lived, transportable, metabolite. Other predators, big-eyed bugs, and antlion larvae were insensitive to this defense. Thus, chemical defenses, too toxic to sequester, can be repurposed for defensive functions through respiration as a form of defensive halitosis, and predators can assist the functional elucidation of herbivore genes.

Insights into the biochemical defence and methylation of the solitary bee Osmia rufa L: A foundation for examining eusociality development.

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Aneta Strachecka

Full Text Available We examined age-related biochemical and histological changes in the fat bodies and hemolymph of Osmia rufa males and females. We analysed solitary bees during diapause, in October and in April; as well as the flying insects following diapause, in May and June. The trophocyte sizes, as well as the numbers of lipid droplets were the greatest at the beginning of diapause. Subsequently, they decreased along with age. Triglyceride and glucose concentrations systematically decreased in fat body cells but increased in the hemolymph from October to June. Concentrations/activities of (enzymatic and non-enzymatic antioxidant and proteolytic systems, as well as phenoloxidase, aspartate aminotransferase, alanine aminotransferase and alkaline phosphatase levels were constant during the diapause, usually lower in the males than the females. Prior to the diapause/overwintering, the concentrations/activities of all the compounds were higher in the fat bodies than in the hemolymph. Later in the spring and in the summer, they increased in the hemolymph and on the body surfaces, while decreasing in the fat bodies. The global DNA methylation levels increased with age. Higher levels were always observed in the males than in the females. The study will promote better understanding of bee evolution and will be useful for the protection and management of solitary bees, with benefits to the environment and agriculture.

Deltamethrin Binding to Triatoma infestans (Hemiptera: Reduviidae) Lipoproteins. Analysis by Solvent Bar Microextraction Coupled to Gas Chromatography.

Science.gov (United States)

Dulbecco, A B; Mijailovsky, S J; Girotti, J R; Juárez, M P

2015-11-01

The binding of deltamethrin (DLM) to the hemipteran Triatoma infestans (Klug) hemolymph lipoproteins was evaluated in vitro. After DLM incubation with the insect hemolymph, lipoproteins were fractioned by ultracentrifugation. DLM binding was analyzed by a microextractive technique-solvent bar microextraction-a solventless methodology to extract DLM from each lipoprotein fraction. This is a novel use of the technique applied to extract an insecticide from an insect fluid. Capillary gas chromatography with microelectron capture detection was used to detect DLM bound by the T. infestans hemolymph lipoproteins and to identify the preferred DLM carrier. We show that Lp and VHDLp I lipoproteins are mainly responsible for DLM transport in T. infestans, both in DLM-resistant and DLM-susceptible bugs. Our results also indicate that DLM amounts transported are not related to DLM susceptibility. © The Authors 2015. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

Immune-physiological aspects of synergy between avermectins and the entomopathogenic fungus Metarhizium robertsii in Colorado potato beetle larvae.

Science.gov (United States)

Tomilova, Oksana G; Kryukov, Vadim Yu; Duisembekov, Bahytzhan A; Yaroslavtseva, Olga N; Tyurin, Maksim V; Kryukova, Natalia A; Skorokhod, Valery; Dubovskiy, Ivan M; Glupov, Viktor V

2016-10-01

The interaction between the entomopathogenic fungus Metarhizium robertsii and natural avermectin metabolites of the actinomycete Streptomyces avermitilis were investigated on Colorado potato beetle larvae. A synergy in the mortality of larvae was detected after simultaneous treatment with half-lethal doses of avermectins (commercial name actarophit) 0.005% and fungus (5×10 5 conidia/ml). The treatment with avermectins led to rapid fungal colonization of the hemolymph. The defense strategies of insects infected by fungus and treated with avermectins and untreated insects were compared to investigate the mechanisms of this synergy. We have shown an increase in hemocytes, especially immunocompetent cells - plasmatocytes and granular cells in the initial stages of mycosis (third day post inoculation). In contrast, avermectins suppressed cellular immunity in hemolymph. Specifically, avermectins dramatically decreased the count of granular cells in larvae infected and uninfected with fungus. Apoptosis inducement and hemocyte necrosis under the influence of avermectins has been shown in vitro as one of the possible reasons for hemocyte mortality. In addition, avermectins enhanced the activity of phenoloxidases in integuments and hemolymph and increased the activity of glutathione-S-transferases activity in the fat body and hemolymph of infected and uninfected larvae, thereby intensifying the development of fungal infection by M. robertsii in Colorado potato beetle larvae. The combination of fungal infection and avermectins constitutes a new perspective for developing multicomponent bioinsecticides. Copyright © 2016 Elsevier Inc. All rights reserved.

High-titer preparation of Bombyx mori nucleopolyhedrovirus (BmNPV displaying recombinant protein in silkworm larvae by size exclusion chromatography and its characterization

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Tanaka Shigeyasu

2009-06-01

Full Text Available Abstract Background Budded baculoviruses are utilized for vaccine, the production of antibody and functional analysis of transmembrane proteins. In this study, we tried to produce and purify the recombinant Bombyx mori nucleopolyhedrovirus (rBmNPV-hPRR that displayed human (prorenin receptor (hPRR connected with FLAG peptide sequence on its own surface. These particles were used for further binding analysis of hPRR to human prorenin. The rBmNPV-hPRR was produced in silkworm larvae and purified from its hemolymph using size exclusion chromatography (SEC. Results A rapid method of BmNPV titer determination in hemolymph was performed using quantitative real-time PCR (Q-PCR. A correlation coefficient of BmNPV determination between end-point dilution and Q-PCR methods was found to be 0.99. rBmNPV-hPRR bacmid-injected silkworm larvae produced recombinant baculovirus of 1.31 × 108 plaque forming unit (pfu in hemolymph, which was 2.8 × 104 times higher than transfection solution in Bm5 cells. Its purification yield by Sephacryl S-1000 SF column chromatography was 264 fold from larval hemolymph at 4 days post-injection (p.i., but 35 or 39 fold at 4.5 or 5 days p.i., respectively. Protein patterns of rBmNPV-hPRR purified at 4 and 5 days were the same and ratio of envelope proteins (76, 45 and 35 kDa to VP39, one of nucleocapsid proteins, increased at 5 days p.i. hPRR was detected in only purified rBmNPV-hPRR at 5 days p.i.. Conclusion The successful purification of rBmNPV-hPRR indicates that baculovirus production using silkworm larvae and its purification from hemolymph by Sephacryl S-1000 SF column chromatography can provide an economical approach in obtaining the purified BmNPV stocks with high titer for large-scale production of hPRR. Also, it can be utilized for further binding analysis and screening of inhibitors of hPRR.

Osmotic and ionic regulation in shore crabs Carcinus maenas inhabiting a tidal estuary

Science.gov (United States)

Winkler, A.; Siebers, D.; Becker, W.

1988-03-01

Shore crabs Carcinus maenas were exposed to salinities fluctuating according to the natural tidal rhythm. To this end they were maintained in net cages positioned in the estuarine waters of the river Elbe. The cages were lifted every hour, and between 8 12 specimens were analyzed for hemolymph concentrations of Na, K, Ca, Mg, and osmolality. The results obtained were compared with the respective data measured in external brackish water. In addition, the specific activity of Na-K-ATPase in a posterior gill was determined. Hemolymph Na and Mg as well as branchial Na-K-ATPase were also determined in crabs collected in the North Sea and the Baltic. The results show that in C. maenas living in salinities fluctuating with the tides by approx. 15‰ S, Na, K and Ca were hyperregulated, and Mg was effectively hyporegulated. The concentrations of all hemolymph ions and the activity of the Na-K-ATPase were kept constant over the whole tidal cycle. In Baltic crabs, Na was effectively hyperregulated and gill Na-K-ATPase was significantly elevated by a factor of ca 2 when compared with North Sea crabs. It is suggested that long-term hyperregulation of Na in constant salinities results from an increased number of Na-K-ATPase molecules which may change by synthesis or degradation following salinity stress. Constant hemolymph levels of hyperregulated Na in crabs inhabiting fluctuating brackish water are accomplished by activation of existing Na-K-ATPase by low Na and inhibition by higher ambient concentrations.

RNA interference reveals allatotropin functioning in larvae and adults of Spodoptera frugiperda (Lepidoptera, Noctuidae

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I.T.E. Hassanien

2014-05-01

Full Text Available The allatotropin of S. frugiperda (Spofr-AT and its cDNA sequence were characterized 10 years ago, but no functional analyses of the peptide are available. Here we used the RNA interference technique to study the effects of Spofr-AT gene suppression on juvenile hormone (JH and ecdysteroid titers in the hemolymph of larvae, virgin and mated females, and of males. Spofr-AT gene silencing in last instar larvae resulted in an increase in the amount of JH III and 20-hydroxyecdysone in the hemolymph of the animals, corresponding to an acceleration of the prepupal commitment and transformation to the pupa. Mated females showed much higher JH titers in their hemolymph than virgins and laid almost twice the number of eggs. Spofr-AT gene silencing in freshly ecdysed females led to a further increase in egg production and oviposition, but had only a minor effect on the hemoylmph JH titer. Mated females contain considerable amounts of JH I and JH II in their hemoylmph, which are thought to be received from males during copulation. To confirm this hypothesis, we measured the amount of JH homologs in the male accessory reproductive glands (MARG before mating and in the bursa copulatrix (BC of the female after mating. MARG contained high amounts of JH I and JH II, which are transferred to the BC during copulation. One day after mating, JH disappeared from the BC and was then found in the hemolymph of the females. In conclusion, Spofr-AT acts as a true allatotropin in larvae and adults of both sexes of the armyworm.

Studies on the extra- and intracellular acid-base status and its role on metabolic depression in the land snail Helix lucorum (L.) during estivation.

Science.gov (United States)

Michaelidis, B

2002-05-01

The aim of the present study was to examine the acid-base status of extra- and intracellular fluids and its possible role on the regulation of the metabolic rate of Helix lucorum during prolonged estivation. For this purpose, the rate of oxygen consumption for active and estivating snails was determined. The acid-base status was also examined in the hemolymph and tissues from active and estivating snails acclimated at 25 degrees C. In addition, the buffer values of hemolymph and tissues were determined in order to examine whether there is a change in the snails during estivation. The rate of oxygen consumption decreased significantly within the 1st 10 days of estivation from 122.51+/-10 microl.g(-1).h(-1) to 25.86+/-5.2 microl.g(-1).h(-1), indicating a marked decrease in metabolic rate. P(CO2)increased within the 1st 20 days of estivation from 13.52+/-0.68 mmHg to 25.09+/-2.05 mmHg, while the pH of hemolymph (pH(e)) decreased from 7.72+/-0.04 to 7.44+/-0.06. The level of bicarbonates decreased in the hemolymph of estivating snails, indicating a metabolic acidosis, which was moderate in extracellular fluids. In contrast to pH(e), the intracellular pH (pH(i)) was maintained in the tissues of estivating H. lucorum, indicating a regulation of pH(i) despite the developed hypercapnia. According to the results presented here, it seems that the timing of pH(e) changes does not correlate with the timing of metabolic rate reduction in estivating H. lucorum.

Honey bee hemocyte profiling by flow cytometry.

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Marringa, William J; Krueger, Michael J; Burritt, Nancy L; Burritt, James B

2014-01-01

Multiple stress factors in honey bees are causing loss of bee colonies worldwide. Several infectious agents of bees are believed to contribute to this problem. The mechanisms of honey bee immunity are not completely understood, in part due to limited information about the types and abundances of hemocytes that help bees resist disease. Our study utilized flow cytometry and microscopy to examine populations of hemolymph particulates in honey bees. We found bee hemolymph includes permeabilized cells, plasmatocytes, and acellular objects that resemble microparticles, listed in order of increasing abundance. The permeabilized cells and plasmatocytes showed unexpected differences with respect to properties of the plasma membrane and labeling with annexin V. Both permeabilized cells and plasmatocytes failed to show measurable mitochondrial membrane potential by flow cytometry using the JC-1 probe. Our results suggest hemolymph particulate populations are dynamic, revealing significant differences when comparing individual hive members, and when comparing colonies exposed to diverse conditions. Shifts in hemocyte populations in bees likely represent changing conditions or metabolic differences of colony members. A better understanding of hemocyte profiles may provide insight into physiological responses of honey bees to stress factors, some of which may be related to colony failure.

Flight metabolism in Panstrongylus megistus (Hemiptera: Reduviidae): the role of carbohydrates and lipids.

Science.gov (United States)

Canavoso, Lilián E; Stariolo, Raúl; Rubiolo, Edilberto R

2003-10-01

The metabolism of lipids and carbohydrates related to flight activity in Panstrongylus megistus was investigated. Insects were subjected to different times of flight under laboratory conditions and changes in total lipids, lipophorin density and carbohydrates were followed in the hemolymph. Lipids and glycogen were also assayed in fat body and flight muscle. In resting insects, hemolymph lipids averaged 3.4 mg/ml and significantly increased after 45 min of flight (8.8 mg/ml, P < 0.001). High-density lipophorin was the sole lipoprotein observed in resting animals. A second fraction with lower density corresponding to low-density lipophorin appeared in insects subjected to flight. Particles from both fractions showed significant differences in diacylglycerol content and size. In resting insects, carbohydrate levels averaged 0.52 mg/ml. They sharply declined more than twofold after 15 min of flight, being undetectable in hemolymph of insects flown for 45 min. Lipid and glycogen from fat body and flight muscle decreased significantly after 45 min of flight. Taken together, the results indicate that P. megistus uses carbohydrates during the initiation of the flight after which, switching fuel for flight from carbohydrates to lipids.

Flight metabolism in Panstrongylus megistus (Hemiptera: Reduviidae: the role of carbohydrates and lipids

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Lilián E Canavoso

2003-10-01

Full Text Available The metabolism of lipids and carbohydrates related to flight activity in Panstrongylus megistus was investigated. Insects were subjected to different times of flight under laboratory conditions and changes in total lipids, lipophorin density and carbohydrates were followed in the hemolymph. Lipids and glycogen were also assayed in fat body and flight muscle. In resting insects, hemolymph lipids averaged 3.4 mg/ml and significantly increased after 45 min of flight (8.8 mg/ml, P < 0.001. High-density lipophorin was the sole lipoprotein observed in resting animals. A second fraction with lower density corresponding to low-density lipophorin appeared in insects subjected to flight. Particles from both fractions showed significant differences in diacylglycerol content and size. In resting insects, carbohydrate levels averaged 0.52 mg/ml. They sharply declined more than twofold after 15 min of flight, being undetectable in hemolymph of insects flown for 45 min. Lipid and glycogen from fat body and flight muscle decreased significantly after 45 min of flight. Taken together, the results indicate that P. megistus uses carbohydrates during the initiation of the flight after which, switching fuel for flight from carbohydrates to lipids.

Inducible immune proteins in the dampwood termite Zootermopsis angusticollis

Science.gov (United States)

Rosengaus, Rebeca B.; Cornelisse, Tara; Guschanski, Katerina; Traniello, James F. A.

2007-01-01

Dampwood termites, Zootermopsis angusticollis (Isoptera: Termopsidae), mount an immune response to resist microbial infection. Here we report on results of a novel analysis that allowed us to electrophoretically assess changes in hemolymph proteins in the same individual before and after exposure to a pathogen. We demonstrate that contact with a sublethal concentration of the entomopathogenic fungus Metarhizium anisopliae (Deuteromycotina:Hypomycetes) induces the production of protective proteins in nymphs, pseudergates (false workers), and soldiers. Termites exposed to an immunizing dosage of fungal conidia consistently showed an enhancement of constitutive proteins (62-85 kDa) in the hemolymph as well as an induction of novel proteins (28-48 kDa) relative to preimmunization levels. No significant differences in protein banding patterns relative to baseline levels in control and naïve termites were observed. Incubating excised and eluted induced proteins produced by immunized pseudergates or immunized soldiers with conidia significantly reduced the germination of the fungus. The fungistatic effect of eluted proteins differed significantly among five colonies examined. Our results show that the upregulation of protective proteins in the hemolymph underscores the in vivo immune response we previously recorded in Z. angusticollis.

A holistic view of dietary carbohydrate utilization in lobster: digestion, postprandial nutrient flux, and metabolism.

Science.gov (United States)

Rodríguez-Viera, Leandro; Perera, Erick; Casuso, Antonio; Perdomo-Morales, Rolando; Gutierrez, Odilia; Scull, Idania; Carrillo, Olimpia; Martos-Sitcha, Juan A; García-Galano, Tsai; Mancera, Juan Miguel

2014-01-01

Crustaceans exhibit a remarkable variation in their feeding habits and food type, but most knowledge on carbohydrate digestion and utilization in this group has come from research on few species. The aim of this study was to make an integrative analysis of dietary carbohydrate utilization in the spiny lobster Panulirus argus. We used complementary methodologies such as different assessments of digestibility, activity measurements of digestive and metabolic enzymes, and post-feeding flux of nutrients and metabolites. Several carbohydrates were well digested by the lobster, but maize starch was less digestible than all other starches studied, and its inclusion in diet affected protein digestibility. Most intense hydrolysis of carbohydrates in the gastric chamber of lobster occurred between 2-6 h after ingestion and afterwards free glucose increased in hemolymph. The inclusion of wheat in diet produced a slow clearance of glucose from the gastric fluid and a gradual increase in hemolymph glucose. More intense hydrolysis of protein in the gastric chamber occurred 6-12 h after ingestion and then amino acids tended to increase in hemolymph. Triglyceride concentration in hemolymph rose earlier in wheat-fed lobsters than in lobsters fed other carbohydrates, but it decreased the most 24 h later. Analyses of metabolite levels and activities of different metabolic enzymes revealed that intermolt lobsters had a low capacity to store and use glycogen, although it was slightly higher in wheat-fed lobsters. Lobsters fed maize and rice diets increased amino acid catabolism, while wheat-fed lobsters exhibited higher utilization of fatty acids. Multivariate analysis confirmed that the type of carbohydrate ingested had a profound effect on overall metabolism. Although we found no evidence of a protein-sparing effect of dietary carbohydrate, differences in the kinetics of their digestion and absorption impacted lobster metabolism determining the fate of other nutrients.

A holistic view of dietary carbohydrate utilization in lobster: digestion, postprandial nutrient flux, and metabolism.

Directory of Open Access Journals (Sweden)

Leandro Rodríguez-Viera

Full Text Available Crustaceans exhibit a remarkable variation in their feeding habits and food type, but most knowledge on carbohydrate digestion and utilization in this group has come from research on few species. The aim of this study was to make an integrative analysis of dietary carbohydrate utilization in the spiny lobster Panulirus argus. We used complementary methodologies such as different assessments of digestibility, activity measurements of digestive and metabolic enzymes, and post-feeding flux of nutrients and metabolites. Several carbohydrates were well digested by the lobster, but maize starch was less digestible than all other starches studied, and its inclusion in diet affected protein digestibility. Most intense hydrolysis of carbohydrates in the gastric chamber of lobster occurred between 2-6 h after ingestion and afterwards free glucose increased in hemolymph. The inclusion of wheat in diet produced a slow clearance of glucose from the gastric fluid and a gradual increase in hemolymph glucose. More intense hydrolysis of protein in the gastric chamber occurred 6-12 h after ingestion and then amino acids tended to increase in hemolymph. Triglyceride concentration in hemolymph rose earlier in wheat-fed lobsters than in lobsters fed other carbohydrates, but it decreased the most 24 h later. Analyses of metabolite levels and activities of different metabolic enzymes revealed that intermolt lobsters had a low capacity to store and use glycogen, although it was slightly higher in wheat-fed lobsters. Lobsters fed maize and rice diets increased amino acid catabolism, while wheat-fed lobsters exhibited higher utilization of fatty acids. Multivariate analysis confirmed that the type of carbohydrate ingested had a profound effect on overall metabolism. Although we found no evidence of a protein-sparing effect of dietary carbohydrate, differences in the kinetics of their digestion and absorption impacted lobster metabolism determining the fate of other

Antioxidant response of the hard shelled mussel Mytilus coruscus exposed to reduced pH and oxygen concentration.

Science.gov (United States)

Sui, Yanming; Hu, Menghong; Shang, Yueyong; Wu, Fangli; Huang, Xizhi; Dupont, Sam; Storch, Daniela; Pörtner, Hans-Otto; Li, Jiale; Lu, Weiqun; Wang, Youji

2017-03-01

Ocean acidification (OA) and hypoxic events are increasing worldwide problems, their interactive effects have not been well clarified, although their co-occurrence is prevalent. The East China Sea (the Yangtze River estuary area) suffers from not only coastal hypoxia but also pH fluctuation, representing an ideal study site to explore the combined effect of OA and hypoxia on marine bivalves. We experimentally evaluated the antioxidant response of the mussel Mytilus coruscus exposed to three pH levels (8.1, 7.7 and 7.3) at two dissolved oxygen (DO) levels (2.0mgL -1 and 6.0mgL -1 ) for 72h. Activities of superoxide dismutase, catalase, glutathione peroxidase, acid phosphatase, and alkaline phosphatase and levels of malondialdehyde were measured in gills and hemolymph. All enzymatic activities in hemolymph and gills followed a similar pattern throughout the experiment duration. Generally, low DO showed greater effects on enzyme activities than elevated CO 2 . Significant interactions between DO, pH and time were only observed at superoxide dismutase and catalase in both tissues. PCA revealed positive relationships between most enzyme activities in both gills and hemolymph with the exception of alkaline phosphatase activity and the level of malondialdehyde in the hemolymph. Overall, our results suggested that decreased pH and low DO induced similar antioxidant responses in the hard shelled mussel, and showed an additive effect on most enzyme activities. The evaluation of multiple environmental stressors, a more realistic scenario than single ones, is crucial to predict the effect of future global changes on coastal species and our results supply some insights on the potential combined effects of reduced pH and DO on marine bivalves. Copyright © 2016 Elsevier Inc. All rights reserved.

Protective Efficacy of a Pseudoalteromonas Strain in European Abalone, Haliotis tuberculata, Infected with Vibrio harveyi ORM4.

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Offret, Clément; Rochard, Vincent; Laguerre, Hélène; Mounier, Jérôme; Huchette, Sylvain; Brillet, Benjamin; Le Chevalier, Patrick; Fleury, Yannick

2018-02-06

The hemolymph of healthy marine invertebrates is known to harbor antibiotic-producing bacteria belonging to the genus Pseudoalteromonas. Such strains are potential probiotics to control infectious diseases in aquaculture. In the present study, we screened a collection of Pseudoalteromonas strains isolated from the hemolymph of oyster and mussel for antimicrobial activity against Vibrio harveyi, a pathogenic species responsible for high mortality in abalone. Subsequently, the protective efficacy of the most active strain named hCg-6 was investigated in abalone culture faced with a Vibrio harveyi ORM4 infection. First, we have controlled the Pseudoalteromonas hCg-6 safety for abalone health. To that end, animals were immersed for 4 h in Pseudoalteromonas hCg-6 suspensions in seawater. The abalone viability was monitored and Pseudoalteromonas hCg-6 was tracked by quantitative-PCR in abalone hemolymph. After immersion, no abalone death occurred while the strain hCg-6 was significantly detected in hemolymph. Therefore, the strain hCg-6 was considered safe for abalone and evaluated for its ability to protect abalone against V. harveyi (injection of 1 × 10 3 Vibrio per animal). A 4-h long immersion of abalone in a seawater suspension of Pseudoalteromonas hCg-6 (1 × 10 6  CFU mL -1 ) prior to infection with Vibrio harveyi significantly improved the abalone viability. Indeed, 15 days post infection, the hCg-6 treatment used increased the abalone survival rate from 16% in untreated animals to 40% in treated abalone. We hypothesized that Pseudoalteromonas hCg-6 antibacterial activity increased the hemomicrobiota shielding effect. In conclusion, Pseudoalteromonas hCg-6 is a promising anti-Vibrio strain for abalone culture.

Cyclomorphosis in Tardigrada: adaptation to environmental constraints

DEFF Research Database (Denmark)

Halberg, Kenneth Agerlin; Persson, Dennis; Ramløv, Hans

2009-01-01

Tardigrades exhibit a remarkable resilience against environmental extremes. In the present study, we investigate mechanisms of survival and physiological adaptations associated with sub-zero temperatures and severe osmotic stress in two commonly found cyclomorphic stages of the marine eutardigrade...... supercooling in both stages, while excluding the presence of physiologically relevant ice-nucleating agents. Experiments on osmotic stress tolerance show that the active stage tolerates the largest range of salinities. Changes in body volume and hemolymph osmolality of active-stage specimens (350-500 microm...... in hemolymph osmolality. At any investigated external salinity, active-stage H. crispae hyper-regulate, indicating a high water turnover and excretion of dilute urine. This is likely a general feature of eutardigrades....

Soft shell clams Mya arenaria with disseminated neoplasia demonstrate reverse transcriptase activity

Science.gov (United States)

House, M.L.; Kim, C.H.; Reno, P.W.

1998-01-01

Disseminated neoplasia (DN), a proliferative cell disorder of the circulatory system of bivalves, was first reported in oysters in 1969. Since that time, the disease has been determined to be transmissible through water-borne exposure, but the etiological agent has not been unequivocally identified. In order to determine if a viral agent, possibly a retrovirus, could be the causative agent of DN, transmission experiments were performed, using both a cell-free filtrate and a sucrose gradient-purified preparation of a cell-free filtrate of DN positive materials. Additionally, a PCR-enhanced reverse transcriptase assay was used to determine if reverse transcriptase was present in tissues or hemolymph from DN positive soft shell clams Mya arenaria. DN was transmitted to healthy clams by injection with whole DN cells, but not with cell-free flitrates prepared from either tissues from DN positive clams, or DN cells. The cell-free preparations from DN-positive tissues and hemolymph having high levels of DN cells in circulation exhibited positive reactions in the PCR-enhanced reverse transcriptase assay. Cell-free preparations of hemolymph from clams having low levels of DN (<0.1% of cells abnormal), hemocytes from normal soft shell clams, and normal soft shell clam tissues did not produce a positive reaction in the PCR enhanced reverse transcriptase assay.

Melatonin in octopus (Octopus vulgaris): tissue distribution, daily changes and relation with serotonin and its acid metabolite.

Science.gov (United States)

Muñoz, José L P; López Patiño, Marcos A; Hermosilla, Consuelo; Conde-Sieira, Marta; Soengas, José L; Rocha, Francisco; Míguez, Jesús M

2011-08-01

Information regarding melatonin production in molluscs is very limited. In this study the presence and daily fluctuations of melatonin levels were investigated in hemolymph, retina and nervous system-related structures in the cephalopod Octopus vulgaris. Adult animals were maintained in captivity under natural photoperiod and killed at different times in a regular daily cycle. Levels of melatonin, serotonin (5-HT) and its acid metabolite (5-hydroxyindole acetic acid, 5-HIAA) in the hemolymph, retina, optic lobe, and cerebral ganglion were assayed by HPLC. Melatonin content fluctuated rhythmically in the retina and hemolymph, peaking at night. In the retina, but not in the other neural tissues, the rhythm was opposite to that of 5-HT, which displayed basal levels at night. Also, 5-HIAA levels in the retina were higher during the night, supporting that rhythmic melatonin production could be linked to diurnal changes in 5-HT degradation. The high levels of melatonin found in the retina point to it as the major source of melatonin in octopus; in addition, a large variation of melatonin content was found in the optic lobe with maximal values at night. All these data suggest that melatonin might play a role in the transduction of the light-dark cycle information for adjustment of rhythmic physiological events in cephalopods.

Pathogen entrapment by transglutaminase--a conserved early innate immune mechanism.

Directory of Open Access Journals (Sweden)

Zhi Wang

2010-02-01

Full Text Available Clotting systems are required in almost all animals to prevent loss of body fluids after injury. Here, we show that despite the risks associated with its systemic activation, clotting is a hitherto little appreciated branch of the immune system. We compared clotting of human blood and insect hemolymph to study the best-conserved component of clotting systems, namely the Drosophila enzyme transglutaminase and its vertebrate homologue Factor XIIIa. Using labelled artificial substrates we observe that transglutaminase activity from both Drosophila hemolymph and human blood accumulates on microbial surfaces, leading to their sequestration into the clot. Using both a human and a natural insect pathogen we provide functional proof for an immune function for transglutaminase (TG. Drosophila larvae with reduced TG levels show increased mortality after septic injury. The same larvae are also more susceptible to a natural infection involving entomopathogenic nematodes and their symbiotic bacteria while neither phagocytosis, phenoloxidase or-as previously shown-the Toll or imd pathway contribute to immunity. These results firmly establish the hemolymph/blood clot as an important effector of early innate immunity, which helps to prevent septic infections. These findings will help to guide further strategies to reduce the damaging effects of clotting and enhance its beneficial contribution to immune reactions.

Botanical profile of bee pollen from the southern coastal region of Bahia, Brazil Perfil botânico do pólen apícola proveniente do litoral sul da Bahia, Brasil

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Marcos da Costa Dórea

2010-09-01

Full Text Available This paper aims to identify the botanical origin of pollen loads collected by Apis mellifera L. in Canavieiras municipality, Bahia state. It provides a list of polliniferous plant species from the Atlantic Forest biome that are important for the development of regional apiculture. Using the acetolysis method, 35 bee-pollen samples were analyzed qualitatively and quantitatively. Results showed that pollen types Elaeis (23.99%, Mimosa pudica (22.78% and Cecropia (13.68% were the most abundant among the samples. These also showed the highest relative frequencies of the material studied and were important pollen sources for bees in the study area.O presente trabalho teve por objetivo identificar a origem botânica das bolotas de pólen coletadas pelas abelhas Apis mellifera L. no município de Canavieiras, Bahia, gerando uma lista de plantas poliníferas pertencentes ao bioma Mata Atlântica e importantes para o desenvolvimento da apicultura regional. Foram analisadas palinologicamente 35 amostras de pólen apícola, utilizando-se o processo de acetólise para, em seguida, serem feitas análises qualitativas e quantitativas. Os resultados revelaram que os tipos polínicos Elaeis (23,99%, Mimosa pudica (22,78% e Cecropia (13,68% foram os mais abundantes no pólen apícola analisado. Estes tipos polínicos também apresentaram as maiores frequências relativas de todo o material estudado e representam importantes plantas fontes de pólen para as abelhas na área estudada.

Shellfish Hemocyte Data from Flow Cytometers

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National Oceanic and Atmospheric Administration, Department of Commerce — Various immune function tests using shellfish hemolymph have been developed to determine shellfish health. These tests including viability, phagocytosis, adhesion,...

Characterization of Ixodes ricinus Fibrinogen-Related Proteins (Ixoderins Discloses Their Function in the Tick Innate Immunity

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Helena Honig Mondekova

2017-12-01

Full Text Available Ticks are important vectors of serious human and animal disease-causing organisms, but their innate immunity can fight invading pathogens and may have the ability to reduce or block transmission to mammalian hosts. Lectins, sugar-binding proteins, can distinguish between self and non-self-oligosaccharide motifs on pathogen surfaces. Although tick hemolymph possesses strong lectin activity, and several lectins have already been isolated and characterized, little is known about the implementation of these molecules in tick immunity. Here, we have described and functionally characterized fibrinogen-related protein (FReP lectins in Ixodes ticks. We have shown that the FReP family contains at least 27 genes (ixoderins, ixo that could, based on phylogenetic and expression analyses, be divided into three groups with differing degrees of expansion. By using RNA interference-mediated gene silencing (RNAi we demonstrated that IXO-A was the main lectin in tick hemolymph. Further, we found that ixoderins were important for phagocytosis of Gram-negative bacteria and yeasts by tick hemocytes and that their expression was upregulated upon injection of microbes, wounding, or after blood feeding. However, although the tick hemocytes could swiftly phagocytose Borrelia afzelii spirochetes, their transmission and burst of infection in mice was not altered. Our results demonstrate that tick ixoderins are crucial immune proteins that work as opsonins in the tick hemolymph, targeting microbes for phagocytosis or lysis.

Optical sensor for measuring American Lobster vitality

International Nuclear Information System (INIS)

Tomassetti, Brian R. A.; Vetelino, John F.

2011-01-01

The vitality of the American Lobster (Homarus americanus) is correlated to the total hemolymph protein (THP) in lobster hemolymph (blood). The standard technique for determining lobster vitality is to draw blood from a lobster and measure THP with a refractometer. This technique is invasive and endangers the lobster's health since blood must be drawn from the lobster. In the present work an optical sensor is developed to measure a lobster's vitality in vivo. It is comprised of a broadband light source, a monochromator, a fiber optic reflection probe, a spectrometer and a computer. This sensor measures protein concentrations by exciting a lobster with 280 nm and 334 nm wavelength light sources and measuring the corresponding absorbance peaks for THP and the fluorescence peak for hemocyanin (Hc), the majority protein in hemolymph. In this work several lobsters are tested. For each lobster, absorbance and fluorescence peaks are measured using the sensor and compared to protein concentrations measured using a refractometer. It is found that the shell thickness and muscle density, which correspond directly to protein concentration and the molting stage of the lobster have a significant effect on the absorbance and fluorescence measurements. It is also found that within specific molting stages, such as pre-molt and post-molt, protein concentration measured with a refractometer correlates linearly to absorbance and fluorescence measurements with the optical sensor.

Optical Sensor for Measuring American Lobster Vitality

Science.gov (United States)

Tomassetti, Brian R. A.; Vetelino, John F.

2011-06-01

The vitality of the American Lobster (Homarus americanus) is correlated to the total hemolymph protein (THP) in lobster hemolymph (blood). The standard technique for determining lobster vitality is to draw blood from a lobster and measure THP with a refractometer. This technique is invasive and endangers the lobster's health since blood must be drawn from the lobster. In the present work an optical sensor is developed to measure a lobster's vitality in vivo. It is comprised of a broadband light source, a monochromator, a fiber optic reflection probe, a spectrometer and a computer. This sensor measures protein concentrations by exciting a lobster with 280 nm and 334 nm wavelength light sources and measuring the corresponding absorbance peaks for THP and the fluorescence peak for hemocyanin (Hc), the majority protein in hemolymph. In this work several lobsters are tested. For each lobster, absorbance and fluorescence peaks are measured using the sensor and compared to protein concentrations measured using a refractometer. It is found that the shell thickness and muscle density, which correspond directly to protein concentration and the molting stage of the lobster have a significant effect on the absorbance and fluorescence measurements. It is also found that within specific molting stages, such as pre-molt and post-molt, protein concentration measured with a refractometer correlates linearly to absorbance and fluorescence measurements with the optical sensor.

POPULATION PHARMACOKINETICS OF ENROFLOXACIN AND ITS METABOLITE CIPROFLOXACIN IN THE GREEN SEA URCHIN (STRONGYLOCENTROTUS DROEBACHIENSIS) FOLLOWING INTRACOELOMIC AND IMMERSION ADMINISTRATION.

Science.gov (United States)

Phillips, Brianne E; Harms, Craig A; Lewbart, Gregory A; Lahner, Lesanna L; Haulena, Martin; Rosenberg, Justin F; Papich, Mark G

2016-03-01

Sea urchin mass mortality events have been attributed to both infectious and noninfectious etiologies. Bacteria, including Vibrio spp. and Pseudoalteromonas spp., have been isolated during specific mortality events. Aquarium collection sea urchins are also subject to bacterial infections and could benefit from antimicrobial treatment, but pharmacokinetic studies have been lacking for this invertebrate group until recently. This study evaluated the pharmacokinetics of enrofloxacin and its active metabolite ciprofloxacin in the green sea urchin (Strongylocentrotus droebachiensis) after intracoelomic injection and medicated bath immersion administration. The utility of a population pharmacokinetic method using nonlinear mixed effects modeling (NLME) was also evaluated. Thirty sea urchins were assigned to either the injection or immersion group. Twelve study animals and three untreated controls were utilized for each administration method: enrofloxacin 10 mg/kg intracoelomic injection or a 6-hr enrofloxacin 10 mg/L immersion. Each animal was sampled four times from 0 to 120 hr. Water samples were collected during immersion treatment and posttreatment time points in both groups. Hemolymph and water sample drug concentrations were analyzed using high-performance liquid chromatography, and pharmacokinetic parameters were determined using an NLME population pharmacokinetic method. Enrofloxacin concentrations were fit to a two-compartment model with first-order input for the intracoelomic injection group. The enrofloxacin elimination half-life (t½), peak hemolymph concentration (CMAX), and area under the curve (AUC) were 38.82 hr, 90.92 μg/ml, and 1,199 hr·μg/ml, respectively. Enrofloxacin was modeled to a one-compartment model with first-order input for the immersion treatment. The enrofloxacin t½, CMAX, and AUC were 33.46 hr, 0.48 μg/ml, and 32.88 hr·μg/ml, respectively. Ciprofloxacin was detected in trace concentrations in all hemolymph samples, indicating

The temporal pattern of vitellogenin synthesis in Drosophila grimshawi

International Nuclear Information System (INIS)

Kambysellis, M.P.; Hatzopoulos, P.; Craddock, E.M.

1989-01-01

The temporal pattern of protein production and, in particular, vitellogenin protein synthesis during the sexual maturation of Drosophila grimshawi females has been studied in vivo by briefly feeding the flies with 35S-methionine and 3H-amino acids. The overall level of incorporation was very low in young flies; it then progressively increased to reach a maximum with the onset of sexual maturity at 13-15 days. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analyses revealed three classes of proteins: those synthesized throughout the age spectrum, which constitute the majority of protein species; proteins synthesized primarily or only in young flies; and proteins synthesized only by the older flies. In this Drosophila species, the three vitellogenins (V1, V2, and V3) appeared to be synthesized in a two-phase pattern. In the first phase, small quantities of V1 and V2 were detected immunologically in the fat body and hemolymph of newly emerged and 1 day-old flies. These proteins did not accumulate in the hemolymph or the ovaries, apparently being unstable proteins. The second phase commenced in early vitellogenesis (7-9 days of age) with synthesis in the fat body of small quantities of V1 and V2, followed by V3 proteins. These proteins were secreted and accumulated in the hemolymph and 24 h later were found in the ovaries. Their quantities increased rapidly and a steady state of synthesis, release into the hemolymph, and uptake by the ovaries was reached by days 13-15. We have estimated that during the steady state of vitellogenin synthesis, a fly can synthesize in 24 h at least 152 micrograms of vitellogenins, which is more than 2% of its body weight, at an average rate of about 6.3 micrograms vitellogenins/h. About 2 micrograms of this are synthesized in the fat body, and about 4 micrograms in the ovaries

Induced thermal stress on serotonin levels in the blue swimmer crab, Portunus pelagicus

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Saravanan Rajendiran

2016-03-01

Full Text Available The temperature of habitat water has a drastic influence on the behavioral, physiological and biochemical mechanisms of crustaceans. Hyperglycemia is a typical response of many aquatic animals to harmful physical and chemical environmental changes. In crustaceans increased circulating crustacean hyperglycemic hormone (CHH and hyperglycemia are reported to occur following exposure to several environmental stress. The biogenic amine, serotonin has been found to modulate the CHH levels and oxidation of serotonin into its metabolites is catalysed by monoamine oxidase. The blue swimmer crab, Portunus pelagicus is a dominant intertidal species utilized throughout the indo-pacific region and is a particularly important species of Palk bay. It has high nutritional value and delicious taste and hence their requirements of capture and cultivation of this species are constantly increasing. This species experiences varying and increasing temperature levels as it resides in an higher intertidal zone of Thondi coast. The present study examines the effect of thermal stress on the levels of serotonin and crustacean hyperglycemic hormone in the hemolymph of P. pelagicus and analyzes the effect of the monoamine oxidase inhibitor, pargyline on serotonin and CHH level after thermal stress. The results showed increased levels of glucose, CHH and serotonin on exposure to 26 °C in control animals. Pargyline injected crabs showed highly significant increase in the levels of CHH and serotonin on every 2 °C increase or decrease in temperature. A greater CHH level of 268.86±2.87 fmol/ml and a greater serotonin level of 177.69±10.10 ng/ml was observed at 24 °C. This could be due to the effect of in maintaining the level of serotonin in the hemolymph and preventing its oxidation, which in turn induces hyperglycemia by releasing CHH into hemolymph. Thus, the study demonstrates the effect of thermal stress on the hemolymph metabolites studied and the role of

Induced thermal stress on serotonin levels in the blue swimmer crab, Portunus pelagicus.

Science.gov (United States)

Rajendiran, Saravanan; Muhammad Iqbal, Beema Mahin; Vasudevan, Sugumar

2016-03-01

The temperature of habitat water has a drastic influence on the behavioral, physiological and biochemical mechanisms of crustaceans. Hyperglycemia is a typical response of many aquatic animals to harmful physical and chemical environmental changes. In crustaceans increased circulating crustacean hyperglycemic hormone (CHH) and hyperglycemia are reported to occur following exposure to several environmental stress. The biogenic amine, serotonin has been found to modulate the CHH levels and oxidation of serotonin into its metabolites is catalysed by monoamine oxidase. The blue swimmer crab, Portunus pelagicus is a dominant intertidal species utilized throughout the indo-pacific region and is a particularly important species of Palk bay. It has high nutritional value and delicious taste and hence their requirements of capture and cultivation of this species are constantly increasing. This species experiences varying and increasing temperature levels as it resides in an higher intertidal zone of Thondi coast. The present study examines the effect of thermal stress on the levels of serotonin and crustacean hyperglycemic hormone in the hemolymph of P. pelagicus and analyzes the effect of the monoamine oxidase inhibitor, pargyline on serotonin and CHH level after thermal stress. The results showed increased levels of glucose, CHH and serotonin on exposure to 26 °C in control animals. Pargyline injected crabs showed highly significant increase in the levels of CHH and serotonin on every 2 °C increase or decrease in temperature. A greater CHH level of 268.86±2.87 fmol/ml and a greater serotonin level of 177.69±10.10 ng/ml was observed at 24 °C. This could be due to the effect of in maintaining the level of serotonin in the hemolymph and preventing its oxidation, which in turn induces hyperglycemia by releasing CHH into hemolymph. Thus, the study demonstrates the effect of thermal stress on the hemolymph metabolites studied and the role of pargyline in elevating the

Target innervation is necessary for neuronal polyploidization in the terrestrial slug Limax.

Science.gov (United States)

Matsuo, Ryota; Yamagishi, Miki; Wakiya, Kyoko; Tanaka, Yoko; Ito, Etsuro

2013-08-01

The brain of gastropod mollusks contains many giant neurons with polyploid genomic DNAs. Such DNAs are generated through repeated DNA endoreplication during body growth. However, it is not known what triggers DNA endoreplication in neurons. There are two possibilities: (1) DNAs are replicated in response to some unknown molecules in the hemolymph that reflect the nutritive status of the animal; or (2) DNAs are replicated in response to some unknown factors that are retrogradely transported through axons from the innervated target organs. We first tested whether hemolymph with rich nutrition could induce DNA endoreplication. We tested whether the transplanted brain exhibits enhanced DNA endoreplication like an endogenous brain does when transplanted into the homocoel of the body of a slug whose body growth is promoted by an increased food supply. However, no enhancement was observed in the frequency of DNA endoreplication when we compared the transplanted brains in the growth-promoted and growth-suppressed host slugs, suggesting that the humoral environment is irrelevant to triggering the body growth-dependent DNA endoreplication. Next, we tested the requirement of target innervation by surgically dissecting a unilateral posterior pedal nerve of an endogenous brain. Substantially lower number of neurons exhibited DNA endoreplication in the pedal ganglion ipsilateral to the dissected nerve. These results support the view that enhanced DNA endoreplication is mediated by target innervation and is not brought about through the direct effect of humoral factors in the hemolymph during body growth. Copyright © 2013 Wiley Periodicals, Inc.

Pharmacokinetics and tissue distribution of enrofloxacin after single intramuscular injection in Pacific white shrimp.

Science.gov (United States)

Fang, X; Zhou, J; Liu, X

2018-02-01

The pharmacokinetic properties and tissue distribution of enrofloxacin (EF) were investigated after single intramuscular (i.m.) dose of 10 mg/kg body weight (b.w.) in Pacific white shrimp at 22 to 25°C. EF and its metabolite ciprofloxacin (CF) were determined by high-performance liquid chromatography. After i.m. administration, EF was absorbed quickly, and the peak of EF concentration (C max ) reached at first time point in hemolymph. The volume of distribution V d(area) of EF was 3.84 L/kg, indicating that the distribution of EF was good. The area under the concentration-time curve (AUC) of EF was 90.1 and 274.2 μg hr/ml in muscle and hepatopancreas, respectively, which was higher than 75.8 μg hr/ml in hemolymph. The EF elimination was slow in muscle and hepatopancreas with the half-life (T 1/2β ) of 52.3 and 75.8 hr, respectively. CF, the mainly metabolite of EF, was detected in hemolymph, muscle and hepatopancreas. The C max was 0.030, 0.013 and 0.218 μg/ml, respectively. Based on a minimum inhibitory concentration (MIC) of 0.006-0.032 μg/ml for susceptible strains, EF i.m. injected at a dose 10 mg/kg could be efficacious against common pathogenic bacteria of Pacific white shrimp. © 2017 John Wiley & Sons Ltd.

Cloning and baculovirus expression of a desiccation stress gene from the beetle, Tenebrio molitor.

Science.gov (United States)

Graham, L A; Bendena, W G; Walker, V K

1996-02-01

The cDNA sequence encoding a novel desiccation stress protein (dsp28) found in the hemolymph of the common yellow mealworm beetle, Tenebrio molitor, has been determined. The sequence encodes a 225 amino acid protein containing a 20 amino acid signal peptide. Dsp28 shows no significant similarity to any known nucleic acid or protein sequence. Levels of dsp28 mRNA were found to increase approx 5-fold following desiccation. Dsp28 cDNA has been cloned into a baculovirus expression vector and the expressed protein was compared to native dsp28. Both dsp28 expressed by recombinant baculovirus and native dsp28 are glycosylated and N-terminally processed. Although dsp28 is induced by cold in addition to desiccation stress, it does not contribute to the freezing point depression (thermal hysteresis) observed in Tenebrio hemolymph.

Long-term in vitro generation of amoebocytes from the Indian horseshoe crab Tachypleus gigas (Muller)

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Joshi, B.; Chatterji, A.; Bhonde, R.

Amoebocyte is the single type of cell circulating in the horseshoe crab hemolymph, which plays a major role in the defense system of the animal. Granules present in these cells are sensitive to nanogram quantities of bacterial endotoxins, which form...

Nitrate binding to Limulus polyphemus subunit type II hemocyanin and its functional implications

NARCIS (Netherlands)

Hazes, B; Magnus, KA; Kalk, KH; Bonaventura, C; Hol, WGJ

1996-01-01

The horseshoe crab, Limulus polyphemus, employs hemocyanin as an oxygen carrier in its hemolymph. This hemocyanin displays cooperative oxygen binding and heterotropic allosteric regulation by protons, chloride ions and divalent cations. Here, we report the crystal structure of Limulus polyphemus

Citogenética de Angiospermas coletadas em Pernambuco: V Cytogenetics of Angiosperms collected in the State of Pernambuco: V

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Andrea Pedrosa

1999-04-01

Full Text Available Foram analisadas 33 espécies, entre nativas e introduzidas, pertencentes a 20 famílias de angiospermas ocorrentes no Estado de Pernambuco. A caracterização cariotípica da maioria das espécies foi baseada no número e morfologia cromossômica, padrão de condensação de cromossomos profásicos e estrutura de núcleo interfásico. Cinco espécies tiveram seus números cromossômicos determinados pela primeira vez, sendo elas: Cereus jamacaru (2n=22, Clitoria fairchildiana (2n=22, Eugenia luschnathiana (2n=22, Licania tomentosa (2n=22 e Spondias tuberosa (n=16. No caso de Licania tomentosa esta é a primeira citação de número cromossômico para o gênero. Das outras 28 espécies, três (Cecropia cf. palmata, 2n=26; Crinum erubescens, 2n=70; e Schinus terebentifolius, 2n=28 apresentaram números cromossômicos diferentes dos registrados previamente na literatura.Thirty three native and introduced species from 20 families of angiosperms collected in the State of Pernambuco were analysed. The karyotype description of the majority of the species was based on chromosome number and morphology, condensation pattern of prophase chromosomes as well as interphase nuclear structure. In five species (Cereus jamacaru, 2n=22; Clitoria fairchildiana, 2n=22; Eugenia luschnathiana, 2n=22; Licania tomentosa, 2n=22; and Spondias tuberosa, n=16 the chromosome number is reported here for the first time. In the case of Licania tomentosa, this is also the first report for the genus. Among the other 28 species, three (Cecropia cf. palmata, 2n=26; Crinum erubescens, 2n=70; and Schinus terebentifolius, 2n=28 showed chromosome numbers different from what has previously been reported.

Detection of Ostreid herpesvirus-1 microvariants in healthy Crassostrea gigas following disease events and their possible role as reservoirs of infection.

Science.gov (United States)

Evans, Olivia; Hick, Paul; Whittington, Richard J

2017-09-01

Ostreid herpesvirus-1 microvariants (OsHV-1) cause severe mortalities in farmed Crassostrea gigas in Europe, New Zealand and Australia. Outbreaks are seasonal, recurring in the warmer months of the year in endemic estuaries. The reference genotype and microvariant genotypes of OsHV-1 have been previously detected in the tissues of apparently healthy adult oysters naturally exposed to OsHV-1 in the field. However, the role of such oysters as reservoirs of infection for subsequent mortality outbreaks remains unclear. The aims of this study were: (1) to identify the optimal sample type to use for the detection of OsHV-1 DNA in apparently healthy C. gigas; and (2) to assess whether live C. gigas maintained on-farm after an OsHV-1 related mortality event remain infected and could act as a reservoir host for subsequent outbreaks. OsHV-1 DNA was detected in the hemolymph, gill, mantle, adductor muscle, gonad and digestive gland of apparently healthy adult oysters. The likelihood of detecting OsHV-1 DNA in hemolymph was equivalent to that in gill and mantle, but the odds of detecting OsHV-1 DNA in hemolymph and gill were more than 8 times that of adductor muscle. Gill had the highest viral loads. Compared to testing whole gill homogenates, testing snippets of the gill improved the detection of OsHV-1 DNA by about four fold. The prevalence of OsHV-1 in gill and mantle was highest after the first season of OsHV-1 exposure; it then declined to low or negligible levels in the same cohorts in subsequent seasons, despite repeated seasonal exposure in monitoring lasting up to 4years. The hemolymph of individually identified oysters was repeatedly sampled over 15months, and OsHV-1 prevalence declined over that time frame in the youngest cohort, which had been exposed to OsHV-1 for the first time at the start of that season. In contrast, the prevalence in two cohorts of older oysters, which had been exposed to OsHV-1 in prior seasons, was consistently low (<10%). Viral loads were

Ultrastructure of book gill development in embryos and first instars of the horseshoe crab Limulus polyphemus L. (Chelicerata, Xiphosura

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Farley Roger D

2012-03-01

Full Text Available Abstract Background The transmission electron microscope (TEM is used for the first time to study the development of book gills in the horseshoe crab. Near the end of the nineteenth century the hypothesis was presented for homology and a common ancestry for horseshoe crab book gills and arachnid book lungs. The present developmental study and the author's recent ones of book gills (SEM and scorpion book lungs (TEM are intended to clarify early histological work and provide new ultrastructural details for further research and for hypotheses about evolutionary history and relationships. Results The observations herein are in agreement with earlier reports that the book gill lamellae are formed by proliferation and evagination of epithelial cells posterior to opisthosomal branchial appendages. A cartilage-like endoskeleton is produced in the base of the opisthosomal appendages. The lamellar precursor cells in the appendage base proliferate, migrate outward and secrete the lamellar cuticle from their apical surface. A series of external, posteriorly-directed lamellae is formed, with each lamella having a central channel for hemolymph and pillar-type space holders formed from cells of the opposed walls. This repeated, page-like pattern results also in water channels (without space holders between the sac-like hemolymph lamellae. Conclusions The developmental observations herein and in an earlier study (TEM of scorpion book lungs show that the lamellae in book gills and book lungs result from some similar activities and features of the precursor epithelial cells: proliferation, migration, alignment and apical/basal polarity with secretion of cuticle from the apical surface and the basal surface in contact with hemolymph. These cellular similarities and the resulting book-like structure suggest a common ancestry, but there are also substantial developmental differences in producing these organs for gas exchange in the different environments, aqueous

Investigating the Effect of Different Treatments with Lactic Acid Bacteria on the Fate of Listeria monocytogenes and Staphylococcus aureus Infection in Galleria mellonella Larvae.

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Grounta, Athena; Harizanis, Paschalis; Mylonakis, Eleftherios; Nychas, George-John E; Panagou, Efstathios Z

2016-01-01

The use of Galleria mellonella as a model host to elucidate microbial pathogenesis and search for novel drugs and therapies has been well appreciated over the past years. However, the effect of microorganisms with functional appeal in the specific host remains scarce. The present study investigates the effect of treatment with selected lactic acid bacteria (LAB) with probiotic potential, as potential protective agents by using live or heat-killed cells at 6 and 24 h prior to infection with Listeria monocytogenes and Staphylococcus aureus or as potential therapeutic agents by using cell-free supernatants (CFS) after infection with the same pathogens. The employed LAB strains were Lactobacillus pentosus B281 and Lactobacillus plantarum B282 (isolated from table olive fermentations) along with Lactobacillus rhamnosus GG (inhabitant of human intestinal tract). Kaplan-Meier survival curves were plotted while the pathogen's persistence in the larval hemolymph was determined by microbiological analysis. It was observed that the time (6 or 24 h) and type (live or heat-killed cells) of challenge period with LAB prior to infection greatly affected the survival of infected larvae. The highest decrease of L. monocytogenes population in the hemolymph was observed in groups challenged for 6 h with heat-killed cells by an average of 1.8 log units compared to non challenged larvae for strains B281 (p 0.0322), B282 (p 0.0325), and LGG (p 0.0356). In the case of S. aureus infection, the population of the pathogen decreased in the hemolymph by 1 log units at 8 h post infection in the groups challenged for 6 h with heat-killed cells of strains B281 (p 0.0161) and B282 (p 0.0096) and by 1.8 log units in groups challenged with heat-killed cells of LGG strain (p 0.0175). Further use of CFS of each LAB strain did not result in any significant prolonged survival but interestingly it resulted in pronounced decrease of L. monocytogenes in the hemolymph at 24 h and 48 h after infection by

Development of two related endoparasitoids in larvae of the diamondback moth, Plutella xylostella (Lepidoptera: Plutellidae)

NARCIS (Netherlands)

Malčická, Mima; Harvey, Jeffrey A.

2015-01-01

We compare the growth and development of two related solitary endoparasitoids (Braconidae, Microgastinae) in different instars (second and third) of the diamondback moth Plutella xylostella. Cotesia vestalis is a well-studied parasitoid whose larvae feed primarily on host hemolymph and fat body

Effects of molting and environmental factors on trace metal body-burdens and hemocyanin concentrations in the American lobster, Homarus americanus.

NARCIS (Netherlands)

Engel, D.W.; Brouwer, M.H.J.; Mercaldo-Allen, R.

2001-01-01

Hemocyanin concentrations in the hemolymph of marine crustacea are dependent on the molt cycle and on environmental conditions. Studies in our laboratories have found that hemocyanin levels in blue crabs are reduced after ecdysis and under conditions of environmental stress (Engel, Brouwer, &

Copper sulphate (CuSO4) toxicity on tissue phosphatases activity and carbohydrates turnover in Achatina fulica.

Science.gov (United States)

Ramalingam, K; Indra, D

2002-04-01

A time course study on the sublethal toxicity of CuSO4 on tissue carbohydrate metabolites level and their phosphatases activity in Achatina fulica revealed differential response. The levels of total carbohydrates and glycogen in the body mass muscle, foot muscle and hemolymph revealed their involvement in the endogenous derivation of energy during stress. The same metabolites in digestive gland revealed its importance to reproduction and development. The lactate accumulated in all the tissues implied the mechanism of CuSO4 toxicosis in the metabolic acidosis. The decrease of pyruvate in foot muscle, body mass muscle and hemolymph inferred the preponderance of glycolysis in energy derivation. In contrast, the pyruvate concentration in digestive gland revealed its differential response in the stress metabolic sequence of changes, as a unique tissue. The lactate/pyruvate ratio and the calcium content in tissues constitute direct evidences for the snails adaptation to toxic stress.

A semiquantitative PCR assay for assessing Perkinsus marinus infections in the eastern oyster, Crassostrea virginica.

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Marsh, A G; Gauthier, J D; Vasta, G R

1995-08-01

A 3.2-kb fragment of Perkinsus marinus DNA was cloned and sequenced. A noncoding domain was identified and targeted for the development of a semiquantitative polymerase chain reaction (PCR) assay for the presence of P. marinus in eastern oyster tissues. The assay involves extracting total DNA from oyster hemolymph and using 1 microgram of that DNA as template in a stringent PCR amplification with oligonucleotide primers that are specific for the P. marinus 3.2-kb fragment. With this assay, we can detect 10 pg of total P. marinus DNA per 1 microgram of oyster hemocyte DNA with ethidium bromide (EtBr) staining of agarose gels, 100 fg total P. marinus DNA with Southern blot autoradiography, and 10 fg of total P. marinus DNA with dot-blot hybridizations. We have used the sensitivity of the PCR assay to develop a method for estimating the level of P. marinus DNA in oyster hemolymph and have successfully applied this technique to gill tissues. Our semiquantitative assay uses a dilution series to essentially titrate the point at which a P. marinus DNA target is no longer amplified in a sample. We refer to this technique as "dilution endpoint" PCR. Using hemocytes obtained by withdrawing a 1-ml sample of hemolymph, this assay provides a nondestructive methodology for rapidly screening large numbers of adult oysters for the presence and quantification of P. marinus infection levels. This technique is applicable to other tissues (gills) and could potentially be applied to DNA extracts of whole larvae or spat.

Malpighian Tubules as Novel Targets for Mosquito Control

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Peter M. Piermarini

2017-01-01

Full Text Available The Malpighian tubules and hindgut are the renal excretory tissues of mosquitoes; they are essential to maintaining hemolymph water and solute homeostasis. Moreover, they make important contributions to detoxifying metabolic wastes and xenobiotics in the hemolymph. We have focused on elucidating the molecular mechanisms of Malpighian tubule function in adult female mosquitoes and developing chemical tools as prototypes for next-generation mosquitocides that would act via a novel mechanism of action (i.e., renal failure. To date, we have targeted inward rectifier potassium (Kir channels expressed in the Malpighian tubules of the yellow fever mosquito Aedes aegypti and malaria mosquito Anopheles gambiae. Inhibition of these channels with small molecules inhibits transepithelial K+ and fluid secretion in Malpighian tubules, leading to a disruption of hemolymph K+ and fluid homeostasis in adult female mosquitoes. In addition, we have used next-generation sequencing to characterize the transcriptome of Malpighian tubules in the Asian tiger mosquito Aedes albopictus, before and after blood meals, to reveal new molecular targets for potentially disrupting Malpighian tubule function. Within 24 h after a blood meal, the Malpighian tubules enhance the mRNA expression of genes encoding mechanisms involved with the detoxification of metabolic wastes produced during blood digestion (e.g., heme, NH3, reactive oxygen species. The development of chemical tools targeting these molecular mechanisms in Malpighian tubules may offer a promising avenue for the development of mosquitocides that are highly-selective against hematophagous females, which are the only life stage that transmits pathogens.

Sepsis and Hemocyte Loss in Honey Bees (Apis mellifera) Infected with Serratia marcescens Strain Sicaria.

Science.gov (United States)

Burritt, Nancy L; Foss, Nicole J; Neeno-Eckwall, Eric C; Church, James O; Hilger, Anna M; Hildebrand, Jacob A; Warshauer, David M; Perna, Nicole T; Burritt, James B

2016-01-01

Global loss of honey bee colonies is threatening the human food supply. Diverse pathogens reduce honey bee hardiness needed to sustain colonies, especially in winter. We isolated a free-living Gram negative bacillus from hemolymph of worker honey bees (Apis mellifera) found separated from winter clusters. In some hives, greater than 90% of the dying bees detached from the winter cluster were found to contain this bacterium in their hemolymph. Throughout the year, the same organism was rarely found in bees engaged in normal hive activities, but was detected in about half of Varroa destructor mites obtained from colonies that housed the septic bees. Flow cytometry of hemolymph from septic bees showed a significant reduction of plasmatocytes and other types of hemocytes. Interpretation of the16S rRNA sequence of the bacterium indicated that it belongs to the Serratia genus of Gram-negative Gammaproteobacteria, which has not previously been implicated as a pathogen of adult honey bees. Complete genome sequence analysis of the bacterium supported its classification as a novel strain of Serratia marcescens, which was designated as S. marcescens strain sicaria (Ss1). When compared with other strains of S. marcescens, Ss1 demonstrated several phenotypic and genetic differences, including 65 genes not previously found in other Serratia genomes. Some of the unique genes we identified in Ss1 were related to those from bacterial insect pathogens and commensals. Recovery of this organism extends a complex pathosphere of agents which may contribute to failure of honey bee colonies.

New functions of arthropod bursicon: inducing deposition and thickening of new cuticle and hemocyte granulation in the blue crab, Callinectes sapidus.

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J Sook Chung

Full Text Available Arthropod growth requires molt-associated changes in softness and stiffness of the cuticle that protects from desiccation, infection and injury. Cuticle hardening in insects depends on the blood-borne hormone, bursicon (Burs, although it has never been determined in hemolymph. Whilst also having Burs, decapod crustaceans reiterate molting many more times during their longer life span and are encased in a calcified exoskeleton, which after molting undergoes similar initial cuticle hardening processes as in insects. We investigated the role of homologous crustacean Burs in cuticular changes and growth in the blue crab, Callinectes sapidus. We found dramatic increases in size and number of Burs cells during development in paired thoracic ganglion complex (TGC neurons with pericardial organs (POs as neurohemal release sites. A skewed expression of Burs β/Burs α mRNA in TGC corresponds to protein contents of identified Burs β homodimer and Burs heterodimer in POs. In hemolymph, Burs is consistently present at ∼21 pM throughout the molt cycle, showing a peak of ∼89 pM at ecdysis. Since initial cuticle hardness determines the degree of molt-associated somatic increment (MSI, we applied recombinant Burs in vitro to cuticle explants of late premolt or early ecdysis. Burs stimulates cuticle thickening and granulation of hemocytes. These findings demonstrate novel cuticle-associated functions of Burs during molting, while the unambiguous and constant presence of Burs in cells and hemolymph throughout the molt cycle and life stages may implicate further functions of its homo- and heterodimer hormone isoforms in immunoprotective defense systems of arthropods.

Molecular epidemiology of Cryptosporidium spp. and Giardia spp. in mussels (Mytilus californianus) and California sea lions (Zalophus californianus) from Central California.

Science.gov (United States)

Adell, A D; Smith, W A; Shapiro, K; Melli, A; Conrad, P A

2014-12-01

Cryptosporidium and Giardia are of public health importance, with recognized transmission through recreational waters. Therefore, both can contaminate marine waters and shellfish, with potential to infect marine mammals in nearshore ecosystems. A 2-year study was conducted to evaluate the presence of Cryptosporidium and Giardia in mussels located at two distinct coastal areas in California, namely, (i) land runoff plume sites and (ii) locations near sea lion haul-out sites, as well as in feces of California sea lions (CSL) (Zalophus californianus) by the use of direct fluorescent antibody (DFA) detection methods and PCR with sequence analysis. In this study, 961 individual mussel hemolymph samples, 54 aliquots of pooled mussel tissue, and 303 CSL fecal samples were screened. Giardia duodenalis assemblages B and D were detected in hemolymph from mussels collected near two land runoff plume sites (Santa Rosa Creek and Carmel River), and assemblages C and D were detected in hemolymph from mussels collected near a sea lion haul-out site (White Rock). These results suggest that mussels are being contaminated by protozoa carried in terrestrial runoff and/or shed in the feces of CSL. Furthermore, low numbers of oocysts and cysts morphologically similar to Cryptosporidium and Giardia, respectively, were detected in CSL fecal samples, suggesting that CSL could be a source and a host of protozoan parasites in coastal environments. The results of this study showed that Cryptosporidium and Giardia spp. from the feces of terrestrial animals and CSL can contaminate mussels and coastal environments. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

Sepsis and Hemocyte Loss in Honey Bees (Apis mellifera) Infected with Serratia marcescens Strain Sicaria

Science.gov (United States)

Burritt, Nancy L.; Foss, Nicole J.; Neeno-Eckwall, Eric C.; Church, James O.; Hildebrand, Jacob A.; Warshauer, David M.; Perna, Nicole T.; Burritt, James B.

2016-01-01

Global loss of honey bee colonies is threatening the human food supply. Diverse pathogens reduce honey bee hardiness needed to sustain colonies, especially in winter. We isolated a free-living Gram negative bacillus from hemolymph of worker honey bees (Apis mellifera) found separated from winter clusters. In some hives, greater than 90% of the dying bees detached from the winter cluster were found to contain this bacterium in their hemolymph. Throughout the year, the same organism was rarely found in bees engaged in normal hive activities, but was detected in about half of Varroa destructor mites obtained from colonies that housed the septic bees. Flow cytometry of hemolymph from septic bees showed a significant reduction of plasmatocytes and other types of hemocytes. Interpretation of the16S rRNA sequence of the bacterium indicated that it belongs to the Serratia genus of Gram-negative Gammaproteobacteria, which has not previously been implicated as a pathogen of adult honey bees. Complete genome sequence analysis of the bacterium supported its classification as a novel strain of Serratia marcescens, which was designated as S. marcescens strain sicaria (Ss1). When compared with other strains of S. marcescens, Ss1 demonstrated several phenotypic and genetic differences, including 65 genes not previously found in other Serratia genomes. Some of the unique genes we identified in Ss1 were related to those from bacterial insect pathogens and commensals. Recovery of this organism extends a complex pathosphere of agents which may contribute to failure of honey bee colonies. PMID:28002470

Primary structure of the oligosaccharide moiety of hemocyanin from the scorpion Androctonus australis

NARCIS (Netherlands)

Vliegenthart, J.F.G.; Debeire, P.; Montreuil, J.; Goyffon, M.; Kuik, J.A. van; Halbeek, H. van

1986-01-01

Hemocyanin, the copper-containing glycoprotein that serves as an oxygen carrier in the hemolymph of some arthropods and molluscs, was obtained from the blood of the scorpion Androctonus australis. Sugar analysis of the glycoprotein revealed that its carbohydrate moiety is of the N-glycosylic type.

Author Details

African Journals Online (AJOL)

Profile of metabolite and biomolecule concentrations in the hemolymph of Bulinus globosus and Bulinus rohlfsi under aestivation and starvation. Abstract · Vol 32, No 2 (2011) - Articles Comparative Studies of two Fresh Water Snail Distributions and Physico-Chemical Parameters in Selected Human Schistosomiasis ...

A whole genome screening and RNA interference identify a juvenile hormone esterase-like gene of the diamondback moth, Plutella xylostella.

Science.gov (United States)

Gu, Xiaojun; Kumar, Sunil; Kim, Eunjin; Kim, Yonggyun

2015-09-01

Juvenile hormone (JH) plays a crucial role in preventing precocious metamorphosis and stimulating reproduction. Thus, its hemolymph titer should be under a tight control. As a negative controller, juvenile hormone esterase (JHE) performs a rapid breakdown of residual JH in the hemolymph during last instar to induce a larval-to-pupal metamorphosis. A whole genome of the diamondback moth (DBM), Plutella xylostella, has been annotated and proposed 11 JHE candidates. Sequence analysis using conserved motifs commonly found in other JHEs proposed a putative JHE (Px004817). Px004817 (64.61 kDa, pI=5.28) exhibited a characteristic JHE expression pattern by showing high peak at the early last instar, at which JHE enzyme activity was also at a maximal level. RNA interference of Px004817 reduced JHE activity and interrupted pupal development with a significant increase of larval period. This study identifies Px004817 as a JHE-like gene of P. xylostella. Copyright © 2015 Elsevier Ltd. All rights reserved.

Multiple functions of the crustacean gill: osmotic/ionic regulation, acid-base balance, ammonia excretion, and bioaccumulation of toxic metals

Science.gov (United States)

Henry, Raymond P.; Lucu, Čedomil; Onken, Horst; Weihrauch, Dirk

2012-01-01

The crustacean gill is a multi-functional organ, and it is the site of a number of physiological processes, including ion transport, which is the basis for hemolymph osmoregulation; acid-base balance; and ammonia excretion. The gill is also the site by which many toxic metals are taken up by aquatic crustaceans, and thus it plays an important role in the toxicology of these species. This review provides a comprehensive overview of the ecology, physiology, biochemistry, and molecular biology of the mechanisms of osmotic and ionic regulation performed by the gill. The current concepts of the mechanisms of ion transport, the structural, biochemical, and molecular bases of systemic physiology, and the history of their development are discussed. The relationship between branchial ion transport and hemolymph acid-base regulation is also treated. In addition, the mechanisms of ammonia transport and excretion across the gill are discussed. And finally, the toxicology of heavy metal accumulation via the gill is reviewed in detail. PMID:23162474

Direct Sensing of Nutrients via a LAT1-like Transporter in Drosophila Insulin-Producing Cells

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Gérard Manière

2016-09-01

Full Text Available Dietary leucine has been suspected to play an important role in insulin release, a hormone that controls satiety and metabolism. The mechanism by which insulin-producing cells (IPCs sense leucine and regulate insulin secretion is still poorly understood. In Drosophila, insulin-like peptides (DILP2 and DILP5 are produced by brain IPCs and are released in the hemolymph after leucine ingestion. Using Ca2+-imaging and ex vivo cultured larval brains, we demonstrate that IPCs can directly sense extracellular leucine levels via minidiscs (MND, a leucine transporter. MND knockdown in IPCs abolished leucine-dependent changes, including loss of DILP2 and DILP5 in IPC bodies, consistent with the idea that MND is necessary for leucine-dependent DILP release. This, in turn, leads to a strong increase in hemolymph sugar levels and reduced growth. GDH knockdown in IPCs also reduced leucine-dependent DILP release, suggesting that nutrient sensing is coupled to the glutamate dehydrogenase pathway.

A comparative study of metabolism and concentration factors of Fe, Cu, Zn, Mn, Co and Mg in Carcinus maenas and Cancer irroratus ovaries during ovogenesis

International Nuclear Information System (INIS)

Martin, J.-L.M.

1975-01-01

Fe, Cu, Zn, Mn, Co, and Mg were analysed in the ovary of Carcinus maenas and Cancer irroratus during ovogenesis. In both ovaries, the relatives rates, expressed as parts per millions as a ratio of wet and dry weight, are the following: Mg>Zn>Fe>Cu>Mn>Co, while in the hemolymph of Cancer irroratus these relative rates are the following: Mg>Cu>Zn>Fe>Mn>Co. Compared to concentrations of these metals in sea water, Mg expected, all metals in the ovary of Cancer irroratus have a concentration factor upper than 1. Compared to the concentration of metals in the hemolymph is, for Fe, Mn, and Co, the concentration factor upper than 1, and for Cu, Zn and Mg, the concentration factor lower than 1. A study of correlations was done between the concentrations of metals considered in pairs, and between the concentrations of metals and the parameters: water content and gonad index [fr

Role of ecdysteroids in the molting and reproductive cycles of the white shrimp, Penaeus vannamei

International Nuclear Information System (INIS)

Chan, S.M.

1989-01-01

The molt cycle was characterized in the white shrimp Penaeus vannamei based upon changes in the morphology of pleopod setae. These characters were used subsequently to determine changes in ecdysteroids and related biosynthetic events during the stages of the molting cycle and in relation to reproduction. Ecdysteroid titers were measured during the molting cycle. They increased during the premolt, decreased at ecdysis and remained minimal through the intermolt. 20-hydroxyecdysone (20-OHE) was the major ecdysteroid present during the premolt. Injection of [ 3 H]ecdysone resulted in label accumulation in the epidermis and hepatopancreas in the form of 20-OHE which was subsequently hydroxylated further to a polar metabolite. [ 3 H]ecdysone was not metabolized by the Y-organ, hemolymph, muscle or intestine. Eyestalk ablation elevated hemolymph ecdysteroid titers and reduced the duration of all molt stages. However, 20-OHE injection into intact animals caused a disproportionate reduction in the durations of only the intermolt and early premolt stages

What are the elements of motivation for acquisition of conditioned taste aversion?

Science.gov (United States)

Mita, Koichi; Okuta, Akiko; Okada, Ryuichi; Hatakeyama, Dai; Otsuka, Emi; Yamagishi, Miki; Morikawa, Mika; Naganuma, Yuki; Fujito, Yutaka; Dyakonova, Varvara; Lukowiak, Ken; Ito, Etsuro

2014-01-01

The pond snail Lymnaea stagnalis is capable of being classically conditioned to avoid food and to consolidate this aversion into a long-term memory (LTM). Previous studies have shown that the length of food deprivation is important for both the acquisition of taste aversion and its consolidation into LTM, which is referred to as conditioned taste aversion (CTA). Here we tested the hypothesis that the hemolymph glucose concentration is an important factor in the learning and memory of CTA. One-day food deprivation resulted in the best learning and memory, whereas more prolonged food deprivation had diminishing effects. Five-day food deprivation resulted in snails incapable of learning or remembering. During this food deprivation period, the hemolymph glucose concentration decreased. If snails were fed for 2days following the 5-day food deprivation, their glucose levels increased significantly and they exhibited both learning and memory, but neither learning nor memory was as good as with the 1-day food-deprived snails. Injection of the snails with insulin to reduce glucose levels resulted in better learning and memory. Insulin is also known to cause a long-term enhancement of synaptic transmission between the feeding-related neurons. On the other hand, injection of glucose into 5-day food-deprived snails did not alter their inability to learn and remember. However, if these snails were fed on sucrose for 3min, they then exhibited learning and memory formation. Our data suggest that hemolymph glucose concentration is an important factor in motivating acquisition of CTA in Lymnaea and that the action of insulin in the brain and the feeding behavior are also important factors. Copyright © 2013 Elsevier Inc. All rights reserved.

Alteration of carbohydrates metabolism and midgut glucose absorption in Gromphadorhina portentosa after subchronic exposure to imidacloprid and fenitrothion.

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Sawczyn, Tomasz; Dolezych, Bogdan; Klosok, Marcin; Augustyniak, Maria; Stygar, Dominika; Buldak, Rafal J; Kukla, Michal; Michalczyk, Katarzyna; Karcz-Socha, Iwona; Zwirska-Korczala, Krystyna

2012-01-01

This study was undertaken to test the hypothesis that following exposure to insecticides, changes take place in the metabolism of carbohydrates and absorption in the midgut of insects. The Madagascar hissing cockroach (Gromphadorhina portentosa) was chosen for the experiment as a model organism, due to it being easy to breed and its relatively large alimentary tract, which was important when preparing the microperfusion midgut bioassay. In each group of cockroaches treated with imidacloprid and fenitrothion, absorption of glucose, expressed as the area under the curve (AUC), was elevated compared to the control group. Glucose in the hemolymph of the examined insects was present in a vestigial amount, often below the threshold of determination, so the determinable carbohydrate indices were: hemolymph trehalose concentration and fat body glycogen content. The level of trehalose found in the hemolymph of insects when exposed to fenitrothion, and irrespective of the level of concentration mixed into food, were significantly lower when comparing to the control samples. Imidacloprid acted analogically with one exception at the concentration of 10 mg·kg(-1) dry food where trehalose concentration did not differ from the control values. Coupling with fat body glycogen concentration was less visible and appeared only at the concentrations of 5 and 10 mg imidacloprid·kg(-1) dry food. As described in this study changes in the sugar distribution and midgut glucose absorption indicate that insects cover the increased energy needs induced by insecticides; also at the gastrointestinal tract level. The result indicates that the midgut glucose absorption parameters could be considered as a non-specific biomarker of insecticide toxicity.

Down-regulation of activity and expression of three transport-related proteins in the gills of the euryhaline green crab, Carcinus maenas, in response to high salinity acclimation.

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Jillette, Nathaniel; Cammack, Lauren; Lowenstein, Margaret; Henry, Raymond P

2011-02-01

The euryhaline green crab, Carcinus maenas, undergoes an annual cycle of salinity exposure, having to adapt to low salinity during its annual spring migration into estuaries, and then having to re-adapt to high salinity when it moves off-shore at the end of summer. Most studies have focused on low salinity acclimation, the activation of osmoregulatory mechanisms, and the induction of transport protein and transport-related enzyme activity and gene expression. In this study we followed the changes in hemolymph osmolality, carbonic anhydrase activity, and mRNA expression of three proteins through a complete cycle of low (15 ppt) and high (32 ppt) salinity acclimation. One week of low salinity acclimation resulted in hemolymph osmoregulation and a four-fold induction of branchial carbonic anhydrase activity. Relative mRNA expression increased for two CA isoforms (CAc 100-fold, and CAg 7-fold) and the α-subunit of the Na/K-ATPase (8-fold). Upon re-exposure to high salinity, hemolymph osmolality increased to 32 ppt acclimated levels by 6 h, and mRNA levels returned to high salinity, baseline levels within 1 week. However, CA activity remained unchanged in response to high salinity exposure for the first week and then gradually declined to baseline levels over 4 weeks. The relative timing of these changes suggests that while whole-organism physiological adaptations and regulation at the gene level can be very rapid, changes at the level of protein expression and turnover are much slower. It is possible that the high metabolic cost of protein synthesis and/or processing could be the underlying reason for long biological life spans of physiologically important proteins. Published by Elsevier Inc.

Investigating the Effect of Different Treatments with Lactic Acid Bacteria on the Fate of Listeria monocytogenes and Staphylococcus aureus Infection in Galleria mellonella Larvae.

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Athena Grounta

Full Text Available The use of Galleria mellonella as a model host to elucidate microbial pathogenesis and search for novel drugs and therapies has been well appreciated over the past years. However, the effect of microorganisms with functional appeal in the specific host remains scarce. The present study investigates the effect of treatment with selected lactic acid bacteria (LAB with probiotic potential, as potential protective agents by using live or heat-killed cells at 6 and 24 h prior to infection with Listeria monocytogenes and Staphylococcus aureus or as potential therapeutic agents by using cell-free supernatants (CFS after infection with the same pathogens. The employed LAB strains were Lactobacillus pentosus B281 and Lactobacillus plantarum B282 (isolated from table olive fermentations along with Lactobacillus rhamnosus GG (inhabitant of human intestinal tract. Kaplan-Meier survival curves were plotted while the pathogen's persistence in the larval hemolymph was determined by microbiological analysis. It was observed that the time (6 or 24 h and type (live or heat-killed cells of challenge period with LAB prior to infection greatly affected the survival of infected larvae. The highest decrease of L. monocytogenes population in the hemolymph was observed in groups challenged for 6 h with heat-killed cells by an average of 1.8 log units compared to non challenged larvae for strains B281 (p 0.0322, B282 (p 0.0325, and LGG (p 0.0356. In the case of S. aureus infection, the population of the pathogen decreased in the hemolymph by 1 log units at 8 h post infection in the groups challenged for 6 h with heat-killed cells of strains B281 (p 0.0161 and B282 (p 0.0096 and by 1.8 log units in groups challenged with heat-killed cells of LGG strain (p 0.0175. Further use of CFS of each LAB strain did not result in any significant prolonged survival but interestingly it resulted in pronounced decrease of L. monocytogenes in the hemolymph at 24 h and 48 h after

NMR assignments of juvenile hormone binding protein in complex with JH III.

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Suzuki, Rintaro; Tase, Akira; Fujimoto, Zui; Shiotsuki, Takahiro; Yamazaki, Toshimasa

2009-06-01

A hemolymph juvenile hormone binding protein (JHBP) shuttles hydrophobic JH, a key hormone in regulation of the insect life cycle, from the site of the JH biosynthesis to the cells of target organs. We report complete NMR chemical shift assignments of Bombyx mori JHBP in the JH III-bound state.

Potencial alelopático de espécies nativas na germinação e crescimento inicial de Lactuca sativa L. (Asteraceae Allelopathic potential of native species in Lactuca sativa L. (Asteraceae germination and initial growth

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Fabiana Maraschin-Silva

2006-03-01

Full Text Available A alelopatia caracteriza-se pelos efeitos danosos ou benéficos sobre o desenvolvimento da vegetação, causados por substâncias químicas produzidas e liberadas para o ambiente por uma planta. Com o objetivo de avaliar o potencial alelopático de espécies brasileiras, foram testados extratos foliares de Cecropia pachystachya Trec. (Urticaceae, Peltophorum dubium (Spreng. Taub. (Fabaceae, Psychotria leiocarpa Cham. & Schltdl (Rubiaceae, Sapium glandulatum (Vell. Pax (Euphorbiaceae e Sorocea bonplandii (Baill. Burg., Lanj. & Boer (Moraceae, utilizando-se bioensaios de germinação e crescimento e alface (Lactuca sativa L. como planta alvo. Nesses bioensaios, foram usados extratos foliares aquosos nas concentrações de 2 e 4%, preparados por maceração estática com água fria e quente. Os extratos das cinco espécies causaram atraso na germinação dos aquênios da alface, bem como efeitos tóxicos no crescimento das plântulas, com redução e enfraquecimento das raízes. Os resultados obtidos mostraram a presença de substâncias químicas inibidoras nos extratos, revelando potencial alelopático para as cinco espécies avaliadas.Allelopathy is characterized by harmful or beneficial effects on vegetation development, caused by chemical substances produced and released into the environment by the plant. Aiming to assess the allelopathic potential of Brazilian species, aqueous leaf extracts of Cecropia pachystachya Trec. (Urticaceae, Peltophorum dubium (Spreng. Taub. (Fabaceae, Psychotria leiocarpa Cham. & Schltdl (Rubiaceae, Sapium glandulatum (Vell. Pax (Euphorbiaceae, and Sorocea bonplandii (Baill. Burger, Lanj. & Boer (Moraceae were tested on lettuce using germination and growth bioassays. In these bioassays, aqueous leaf extracts were used at concentrations of 2 and 4%, prepared by static maceration with cold and hot water. The five species extracts delayed lettuce germination and produced toxic effects on seedling growth, with root

Espectro polínico de amostras de mel de Apis mellifera L., coletadas na Bahia Pollen spectrum of samples of Apis mellifera L., honey collected in Bahia State, Brazil

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AUGUSTA CAROLINA DE CAMARGO CARMELLO MORETI

2000-01-01

Full Text Available O espectro polínico encontrado em amostras de mel provenientes de seis municípios do Estado da Bahia foi analisado com o objetivo de contribuir para o conhecimento da flora visitada por Apis mellifera L. 1758 (Hymenoptera: ApidaeA identificação dos tipos polínicos foi feita por meio de literatura especializada e de informações de campo. Foram encontrados 43 tipos de pólen, sendo consideradas como dominantes as espécies Eucalyptus sp. (Myrtaceae, Mimosa verrucosa Benth. (Mimosaceae, M. scabrella Benth. (Mimosaceae e Bauhinia sp. (Caesalpiniaceae. Os tipos acessórios foram Alternanthera ficoidea R.Br. (Amaranthaceae, tipo Compositae (Asteraceae e Cecropia sp. (Moraceae. Destacou-se a participação de Mimosa sp. (Mimosaceae e de outras espécies silvestres na composição dos méis nos municípios considerados, registrando-se ainda a espécie Eucalyptus sp. (Myrtaceae como uma das principais fontes de alimento das abelhas dessa região.Pollen spectrum found in honey samples from six localities of Bahia State, Brazil, was analyzed with the objective to contribute for the knowledge of the plants used by Apis mellifera L., 1758 (Hymenoptera: Apidae. The identification of the pollen types was made by specialized literature and by field information. Two hundred pollen grains were studied in order to determine the percentage and the occurrence of each type. Forty three pollen types were identified, being considered as the predominant types Eucalyptus sp. (Myrtaceae, Mimosa verrucosa Benth. (Mimosaceae, M. scabrella Benth. (Mimosaceae and Bauhinia sp. (Caesalpiniaceae. The accessory pollen types were Alternanthera ficoidea R.Br. (Amaranthaceae, Compositae type (Asteraceae and Cecropia sp. (Moraceae. It is intense the participation of Mimosa sp. (Mimosaceae and other wild species in the honey composition of the six localities considered. Eucalyptus sp. (Myrtaceae is one of the dominant sources of bee food in some regions of Bahia State.

Ethnomedicines used in Trinidad and Tobago for urinary problems and diabetes mellitus.

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Lans, Cheryl A

2006-10-13

This paper is based on ethnobotanical interviews conducted from 1996-2000 in Trinidad and Tobago with thirty male and female respondents. A non-experimental validation was conducted on the plants used for urinary problems and diabetes mellitus: This is a preliminary step to establish that the plants used are safe or effective, to help direct clinical trials, and to inform Caribbean physicians of the plants' known properties to avoid counter-prescribing. The following plants are used to treat diabetes: Antigonon leptopus, Bidens alba, Bidens pilosa, Bixa orellana, Bontia daphnoides, Carica papaya, Catharanthus roseus, Cocos nucifera, Gomphrena globosa, Laportea aestuans, Momordica charantia, Morus alba, Phyllanthus urinaria and Spiranthes acaulis. Apium graviolens is used as a heart tonic and for low blood pressure. Bixa orellana, Bontia daphnoides, Cuscuta americana and Gomphrena globosa are used for jaundice. The following plants are used for hypertension: Aloe vera, Annona muricata, Artocarpus altilis, Bixa orellana, Bidens alba, Bidens pilosa, Bonta daphnoides, Carica papaya, Cecropia peltata, Citrus paradisi, Cola nitida, Crescentia cujete, Gomphrena globosa, Hibiscus sabdariffa, Kalanchoe pinnata, Morus alba, Nopalea cochinellifera, Ocimum campechianum, Passiflora quadrangularis, Persea americana and Tamarindus indicus. The plants used for kidney problems are Theobroma cacao, Chamaesyce hirta, Flemingia strobilifera, Peperomia rotundifolia, Petiveria alliacea, Nopalea cochinellifera, Apium graveolens, Cynodon dactylon, Eleusine indica, Gomphrena globosa, Pityrogramma calomelanos and Vetiveria zizanioides. Plants are also used for gall stones and for cooling. Chamaesyce hirta, Cissus verticillata, Kalanchoe pinnata, Peperomia spp., Portulaca oleraceae, Scoparia dulcis, and Zea mays have sufficient evidence to support their traditional use for urinary problems, "cooling" and high cholesterol. Eggplant extract as a hypocholesterolemic agent has some support but

Vibrio cholerae interactions with Mytilus galloprovincialis hemocytes mediated by serum components.

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Laura eCanesi

2013-12-01

Full Text Available Edible bivalves (e.g., mussels, oysters can accumulate large amount of bacteria in their tissues and act as passive carriers of pathogens to humans. Bacterial persistence inside bivalves depends, at least in part, on hemolymph anti-bacterial activity that is exerted by both serum soluble factors and phagocytic cells (i.e., the hemocytes. It was previously shown that Mytilus galloprovincialis hemolymph serum contains opsonins that mediate D-mannose-sensitive interactions between hemocytes and V. cholerae O1 El Tor bacteria that carry the Mannose–Sensitive Hemagglutinin (MSHA. These opsonins enhance phagocytosis and killing of vibrios by facilitating their binding to hemocytes. Since V. cholerae strains not carrying the MSHA ligand (O1 classical, non O1/O139 are present in coastal water and can be entrapped by mussels, we studied whether in mussel serum, in addition to opsonins directed towards MSHA, other components can mediate opsonization of these bacteria. By comparing interactions of O1 classical and non O1/O139 strains with hemocytes in ASW and serum, it was found that M. galloprovincialis serum contains components that increase by at approximately two fold their adhesion to, association with and killing by hemocytes. Experiments conducted with high and low molecular mass fractions obtained by serum ultrafiltration indicated that these compounds have molecular mass higher than 5000 Da. Serum exposure to high temperature (80°C abolished its opsonizing capability suggesting that the involved serum active components are of protein nature. Further studies are needed to define the chemical properties and specificity of both the involved bacterial ligands and hemolymph opsonins. This information will be central not only to better understand V. cholerae ecology, but also to improve current bivalve depuration practices and properly protect human health.

The angiotensin-converting enzyme (ACE) gene family of Bombyx mori.

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Yan, Hai-Yan; Mita, Kazuei; Zhao, Xia; Tanaka, Yoshikazu; Moriyama, Minoru; Wang, Huabin; Iwanaga, Masashi; Kawasaki, Hideki

2017-04-15

We previously reported regarding an ecdysone-inducible angiotensin-converting enzyme (ACE) gene. We found another four ACE genes in the Bombyx genome. The present study was undertaken to clarify the evolutionally changed function of the ACE of Bombyx mori. Core regions of deduced amino acid sequences of ACE genes were compared with those of other insect ACE genes. Five Bombyx genes have the conserved Zn 2+ -binding-site motif (HEXXH); however, BmAcer4 has only one and BmAcer3 has no catalytic ligand. BmAcer1 and BmAcer2 were expressed in several organs. BmAcer3 was expressed in testes, and BmAcer4 and BmAcer5 were expressed in compound eyes; however, the transcription levels of these three genes were very low. Quantitative RT-PCR and Western analysis were conducted to determine the tissue distribution and developmental expression of BmAcer1and BmAcer2. Transcripts of BmAcer1 and BmAcer2 were found in the reproductive organs during the larval and pupal stages. BmAcer1 was dominant in fat bodies during the feeding stage and showed high expression in the epidermis, wing discs, and pupal wing tissues after the wandering stage. Its expression patterns in epidermis, wing discs, and wing tissues resembled the hemolymph ecdysteroid titer in the larval and pupal stages. Acer1 was observed in the hemolymph at all stages, appearing to be the source of it are fat bodies, wings, and epidermis, and functioning after being secreted into the hemolymph. BmAcer2 was abundant in the midgut during the feeding stage and after the wandering stage and in silk glands after the pupal stage. We conclude that the evolution of BmAcer occurred through duplication, and, thereafter, functional diversification developed. Copyright © 2017 Elsevier B.V. All rights reserved.

On the Ultrastructure and Function of Rhogocytes from the Pond Snail Lymnaea stagnalis.

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Maria Kokkinopoulou

Full Text Available Rhogocytes, also termed "pore cells", occur as solitary or clustered cells in the connective tissue of gastropod molluscs. Rhogocytes possess an enveloping lamina of extracellular matrix and enigmatic extracellular lacunae bridged by cytoplasmic bars that form 20 nm diaphragmatic slits likely to act as a molecular sieve. Recent papers highlight the embryogenesis and ultrastructure of these cells, and their role in heavy metal detoxification. Rhogocytes are the site of hemocyanin or hemoglobin biosynthesis in gastropods. Based on electron microscopy, we recently proposed a possible pathway of hemoglobin exocytosis through the slit apparatus, and provided molecular evidence of a common phylogenetic origin of molluscan rhogocytes, insect nephrocytes and vertebrate podocytes. However, the previously proposed secretion mode of the respiratory proteins into the hemolymph is still rather hypothetical, and the possible role of rhogocytes in detoxification requires additional data. Although our previous study on rhogocytes of the red-blooded (hemoglobin-containing freshwater snail Biomphalaria glabrata provided much new information, a disadvantage was that the hemoglobin molecules were not unequivocally defined in the electron microscope. This made it difficult to trace the exocytosis pathway of this protein. Therefore, we have now performed a similar study on the rhogocytes of the blue-blooded (hemocyanin-containing freshwater snail Lymnaea stagnalis. The intracellular hemocyanin could be identified in the electron microscope, either as individual molecules or as pseudo-crystalline arrays. Based on 3D-electron microscopy, and supplemented by in situ hybridization, immunocytochemistry and stress response experiments, we provide here additional details on the structure and hemocyanin biosynthesis of rhogocytes, and on their response in animals under cadmium and starvation stress. Moreover, we present an advanced model on the release of synthesized

Synthesis and binding affinity of an iodinated juvenile hormone

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Prestwich, G.D.; Eng, W.S.; Robles, S.; Vogt, R.G.; Wisniewski, J.R.; Wawrzenczyk, C.

1988-01-25

The synthesis of the first iodinated juvenile hormone (JH) in enantiomerically enriched form is reported. This chiral compound, 12-iodo-JH I, has an iodine atom replacing a methyl group of the natural insect juvenile hormone, JH I, which is important in regulating morphogenesis and reproduction in the Lepidoptera. The unlabeled compound shows approximately 10% of the relative binding affinity for the larval hemolymph JH binding protein (JHBP) of Manduca sexta, which specifically binds natural /sup 3/H-10R,11S-JH I (labeled at 58 Ci/mmol) with a KD of 8 X 10(-8) M. It is also approximately one-tenth as biologically active as JH I in the black Manduca and epidermal commitment assays. The 12-hydroxy and 12-oxo compounds are poor competitors and are also biologically inactive. The radioiodinated (/sup 125/I)12-iodo-JH I can be prepared in low yield at greater than 2500 Ci/mmol by nucleophilic displacement using no-carrier-added /sup 125/I-labeled sodium iodide in acetone; however, synthesis using sodium iodide carrier to give the approximately 50 Ci/mmol radioiodinated ligand proceeds in higher radiochemical yield with fewer by-products and provides a radioligand which is more readily handled in binding assays. The KD of (/sup 125/I)12-iodo-JH I was determined for hemolymph JHBP of three insects: M. sexta, 795 nM; Galleria mellonella, 47 nM; Locusta migratoria, 77 nM. The selectivity of 12-iodo-JH I for the 32-kDa JHBP of M. sexta was demonstrated by direct autoradiography of a native polyacrylamide gel electrophoresis gel of larval hemolymph incubated with the radioiodinated ligand. Thus, the in vitro and in vivo activity of 12-iodo-JH I indicate that it can serve as an important new gamma-emitting probe in the search for JH receptor proteins in target tissues.

Massively parallel amplicon sequencing reveals isotype-specific variability of antimicrobial peptide transcripts in Mytilus galloprovincialis.

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Umberto Rosani

Full Text Available BACKGROUND: Effective innate responses against potential pathogens are essential in the living world and possibly contributed to the evolutionary success of invertebrates. Taken together, antimicrobial peptide (AMP precursors of defensin, mytilin, myticin and mytimycin can represent about 40% of the hemocyte transcriptome in mussels injected with viral-like and bacterial preparations, and unique profiles of myticin C variants are expressed in single mussels. Based on amplicon pyrosequencing, we have ascertained and compared the natural and Vibrio-induced diversity of AMP transcripts in mussel hemocytes from three European regions. METHODOLOGY/PRINCIPAL FINDINGS: Hemolymph was collected from mussels farmed in the coastal regions of Palavas (France, Vigo (Spain and Venice (Italy. To represent the AMP families known in M. galloprovincialis, nine transcript sequences have been selected, amplified from hemocyte RNA and subjected to pyrosequencing. Hemolymph from farmed (offshore and wild (lagoon Venice mussels, both injected with 10(7 Vibrio cells, were similarly processed. Amplicon pyrosequencing emphasized the AMP transcript diversity, with Single Nucleotide Changes (SNC minimal for mytilin B/C and maximal for arthropod-like defensin and myticin C. Ratio of non-synonymous vs. synonymous changes also greatly differed between AMP isotypes. Overall, each amplicon revealed similar levels of nucleotidic variation across geographical regions, with two main sequence patterns confirmed for mytimycin and no substantial changes after immunostimulation. CONCLUSIONS/SIGNIFICANCE: Barcoding and bidirectional pyrosequencing allowed us to map and compare the transcript diversity of known mussel AMPs. Though most of the genuine cds variation was common to the analyzed samples we could estimate from 9 to 106 peptide variants in hemolymph pools representing 100 mussels, depending on the AMP isoform and sampling site. In this study, no prevailing SNC patterns related

New method for control of biological fouling in pipelines based on elevated partial pressures of carbon dioxide or combustion exhaust gases.

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B.J. Watten; R.E. Sears; R.F. Bumgardner [U.S. Geological Survey - Leetown Science Center, Kearneysville, WV (United States)

2005-07-01

Zebra mussels (Dreissena polymorpha) and Asian clams (Corbicula fluminea) are major macro fouling species of water conduits used in industrial and power station raw water/condensor systems. Control typically involves manual scraping and use of thermal treatment, deoxygenation or biocides. The costs of control and system damage resulting from fouling in the United States alone have been estimated at $1 billion/year. There is a pressing need for new economical and safe control strategies. Aquatic species in general are intolerant to increases in PCO{sub 2} given its effect on water, blood and hemolymph pH. These species are also sensitive to increases in elevated total dissolved gas pressure. The gas bubble disease that develops following exposure can, as with CO{sub 2} exposure, lead to mortality. We are exploiting this sensitivity by developing a control method based on manipulation of PCO{sub 2} or power plant exhaust gas (CO{sub 2}, 14%; SO{sub 2}, 0.3%), i.e., the supersaturation of hemolymph and tissues with gas followed by an induced (short term) pressure release designed to induce formation of gas emboli. SO{sub 2} present in stack gas acts with CO{sub 2} to reduce water and hemolymph pH so as to reduce required exposure periods. System effluents are degassed, CO{sub 2} recovered and pH adjusted if needed with alkaline reagents. Test results indicate the new process is effective at controlling C. fluminea as well as other target species, including crustaceans and fish. Required exposure periods (LT50) are short and decrease with increasing gas supersaturation levels. Gas recovery and reuse methods developed have reduced gas requirements by 85% making the method attractive economically and environmentally. 40 refs., 8 figs., 1 tab.

Chronic exposure to soil salinity in terrestrial species: Does plasticity and underlying physiology differ among specialized ground-dwelling spiders?

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Renault, D; Puzin, C; Foucreau, N; Bouchereau, A; Pétillon, J

2016-07-01

In salt marshes, the alternation of low and high tides entails rapid shifts of submersion and aerial exposure for terrestrial communities. In these intertidal environments, terrestrial species have to deal with an osmotic loss in body water content and an increase in sodium chloride concentration when salt load increases. In salt marshes, spiders represent an abundant arthropod group, whose physiological ecology in response to variations of soil salinity must be further investigated. In this study, we compared the effect of salinity on the survival and physiology of three species of Lycosidae; two salt marsh species (Arctosa fulvolineata and Pardosa purbeckensis) and one forest species (P. saltans). Spiders were individually exposed at three salinity conditions (0‰, 35‰ and 70‰) and survival, changes in body water content, hemolymph ions (Na(+), Ca(2+), Mg(2+), K(+); ICP-MS technique) and metabolites (mainly amino acids, polyols, sugars; LC and GC techniques) were assessed. The survival of the forest species P. saltans was very quickly hampered at moderate and high salinities. In this spider, variations of hemolymph ions and metabolites revealed a quick loss of physiological homeostasis and a rapid salt-induced dehydration of the specimens. Conversely, high survival durations were measured in the two salt-marsh spiders, and more particularly in A. fulvolineata. In both P. purbeckensis and A. fulvolineata, the proportion of Na(+), Ca(2+), Mg(2+), K(+) remained constant at the three experimental conditions. Accumulation of hemolymph Na(+) and amino acids (mainly glutamine and proline) demonstrated stronger osmoregulatory capacities in these salt-marsh resident spiders. To conclude, even if phylogenetically close (belonging to the same, monophyletic, family), we found different physiological capacities to cope with salt load among the three tested spider species. Nevertheless, physiological responses to salinity were highly consistent with the realized

Proteomic Analysis of Interaction between a Plant Virus and Its Vector Insect Reveals New Functions of Hemipteran Cuticular Protein.

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Liu, Wenwen; Gray, Stewart; Huo, Yan; Li, Li; Wei, Taiyun; Wang, Xifeng

2015-08-01

Numerous viruses can be transmitted by their corresponding vector insects; however, the molecular mechanisms enabling virus transmission by vector insects have been poorly understood, especially the identity of vector components interacting with the virus. Here, we used the yeast two-hybrid system to study proteomic interactions of a plant virus (Rice stripe virus, RSV, genus Tenuivirus) with its vector insect, small brown planthopper (Laodelphax striatellus). Sixty-six proteins of L. striatellus that interacted with the nucleocapsid protein (pc3) of RSV were identified. A virus-insect interaction network, constructed for pc3 and 29 protein homologs of Drosophila melanogaster, suggested that nine proteins might directly interact with pc3. Of the 66 proteins, five (atlasin, a novel cuticular protein, jagunal, NAC domain protein, and vitellogenin) were most likely to be involved in viral movement, replication, and transovarial transmission. This work also provides evidence that the novel cuticular protein, CPR1, from L. striatellus is essential for RSV transmission by its vector insect. CPR1 binds the nucleocapsid protein (pc3) of RSV both in vivo and in vitro and colocalizes with RSV in the hemocytes of L. striatellus. Knockdown of CPR1 transcription using RNA interference resulted in a decrease in the concentration of RSV in the hemolymph, salivary glands and in viral transmission efficiency. These data suggest that CPR1 binds RSV in the insect and stabilizes the viral concentration in the hemolymph, perhaps to protect the virus or to help move the virus to the salivary tissues. Our studies provide direct experimental evidence that viruses can use existing vector proteins to aid their survival in the hemolymph. Identifying these putative vector molecules should lead to a better understanding of the interactions between viruses and vector insects. © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

Cytochemistry of fat body trophocytes and ovaries of workers and queens of Melipona quadrifasciata anthidioides (Hymenoptera: Apidae: Meliponini) during vitellogenesis.

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Paes-de-Oliveira, Vagner Tadeu; Roat, Thaisa Cristina; Berger, Bruno; Da Cruz-Landim, Carminda

2012-12-01

The fat body (FB) of insects is where yolk proteins are synthesized. Therefore, relationships between the FB and oogenesis were studied in nurse workers, virgins, and physogastric queens of Melipona quadrifasciata anthidioides, a stingless bee in which the workers produce and lay eggs while provisioning brood cells. The relationships between FB and oogenesis, as well as the routes of materials from hemolymph to the oocytes, were studied through the cytochemical detection of lipids by osmium imidazole (OI), carbohydrates by ruthenium red (RR) and basic proteins by ammoniacal silver (AS). The results show differences in the presence of the studied materials in FB trophocytes and ovary of the classes of females studied and oogenesis phases. Material that tested positive for the treatments was detected among the classes of individuals studied in both, trophocytes and oocytes, and in the route of those materials from hemolymph to the oocytes. The differences found among the individual classes indicate relationships with the nutrition and adaptation to the parsimonious use of nutrients in the metabolism of reproduction. Copyright © 2012 Wiley Periodicals, Inc.

Evidence of the Presence of Thyroid Hormones in Achatina fulica Snails

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DANILO LUSTRINO

Full Text Available ABSTRACT The objective of this study was to identify thyroid hormones and to examine their putative site of synthesis in Achatina fulica snails. For this purpose, radioimmunoassays were performed for T3 and T4 before and after long starvation with or without hemolymph deproteinization. Sodium/iodide symporter activity in vivo was analyzed through 125I administration with and without KClO4 pretreatment. Only T4 was detected, and its concentration decreased due to starvation or deproteinization. However, high-performance liquid chromatography analysis also showed the presence of T2 and T3 apart from T4, but rT3 was not detected in the A. fulica hemolymph. The sodium/iodide symporter activity was greater in cerebral ganglia than digestive gland, but KClO4 treatment did not inhibit iodide uptake in any of the tissues analyzed. Altogether, our data confirm for the first time the presence of thyroid hormones in A. fulica snails and suggest their participation in the metabolism control in this species, although the putative site of hormone biosynthesis remains to be elucidated.

Evidence of the Presence of Thyroid Hormones in Achatina fulica Snails.

Science.gov (United States)

Lustrino, Danilo; Silva, Alba C M; Araujo, Iracema G; Tunholi, Victor M; Tunholi-Alves, Vinícius M; Castro, Rosane N; Carvalho, Denise P; Pinheiro, Jairo; Marassi, Michelle P

2017-01-01

The objective of this study was to identify thyroid hormones and to examine their putative site of synthesis in Achatina fulica snails. For this purpose, radioimmunoassays were performed for T3 and T4 before and after long starvation with or without hemolymph deproteinization. Sodium/iodide symporter activity in vivo was analyzed through 125I administration with and without KClO4 pretreatment. Only T4 was detected, and its concentration decreased due to starvation or deproteinization. However, high-performance liquid chromatography analysis also showed the presence of T2 and T3 apart from T4, but rT3 was not detected in the A. fulica hemolymph. The sodium/iodide symporter activity was greater in cerebral ganglia than digestive gland, but KClO4 treatment did not inhibit iodide uptake in any of the tissues analyzed. Altogether, our data confirm for the first time the presence of thyroid hormones in A. fulica snails and suggest their participation in the metabolism control in this species, although the putative site of hormone biosynthesis remains to be elucidated.

Virus present in the reproductive tract of asymptomatic drones of honey bee (Apis mellifera l.), and possible infection of queen during mating.

Science.gov (United States)

Da Cruz-Landim, Carminda; Roat, Thaisa C; Fernadez, Fernanda C

2012-07-01

Virus particles and viral inclusions were detected by transmission electron microscopy examination of sections of the seminal vesicles and mucus gland of asymptomatic young drones from colonies of Apis mellifera lightly infested by Varroa mite. In the mucus gland the infection was found in the muscular sheath and epithelium, while in the seminal vesicle in cells of the outer serosa. Isolated viral particles were also observed in the hemolymph occupying the intercellular spaces of the muscular sheath fibers. In the muscle the virus appeared as polygonal crystalloid inclusions, while in the epithelium mainly inside cytoplasmic vesicles. The infected cells apparently are not damaged. The virus particles are present in the hemolymph and forming more mature structures, as crystalloids, in the muscle. This suggests that the virus is liberated in the body fluid and infects the tissues penetrating the cells through endocytosis. The presence of virus in mucus gland epithelial vesicles raise the possibility of its transference to the gland secretion and therefore, to the semen. Copyright © 2012 Wiley Periodicals, Inc.

Resistance of particleboard panels made of agricultural residues and bonded with synthetic resins or PVC plastic to wood-rotting fungi

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Divino Eterno Teixeira

2009-12-01

Full Text Available This study aims to evaluate the resistance of three types of particleboard panel to biodeterioration, two of which bonded with synthetic resins and one bonded with PVC plastic. Composite panels were made using sugar cane straw particles as raw material which were bonded together with urea-formaldehyde (UF, tannin-formaldehyde (TANI and PVC plastic (PVC resins. Decay tests were performed following procedures outlined in the ASTM D2017-81/1994 standard, whereby sample specimens were subjected to attack by white rot fungus Trametes versicolor and brown rot fungus Gloeophyllum trabeum using pine (Pinus sp. and embaúba (Cecropia sp. as reference timber. Panels bonded with PVC resin were rated ‘resistant’ to attack by both fungi while those bonded with UF and TANI resins were rated ‘slightly resistant’ to their attack.

Hemelipoglycoprotein from the ornate sheep tick, Dermacentor marginatus: structural and functional characterization

Czech Academy of Sciences Publication Activity Database

Dupejová, J.; Štěrba, Ján; Vancová, Marie; Grubhoffer, Libor

2011-01-01

Roč. 4, January (2011), s. 4 ISSN 1756-3305 R&D Projects: GA MŠk(CZ) LC06009; GA AV ČR KJB600960906 Institutional research plan: CEZ:AV0Z60220518 Keywords : ORNITHODOROS PARKERI * BOOPHILUS MICROPLUS * MASS-SPECTROMETRY * CATTLE TICK * SOFT TICK * DORIN M * PROTEINS * HEMOLYMPH * VITELLOGENIN * LIPOPROTEIN Subject RIV: EA - Cell Biology Impact factor: 2.937, year: 2011

The Drosophila chitinase-like protein IDGF3 is involved in protection against nematodes and woung healing

Czech Academy of Sciences Publication Activity Database

Kučerová, Lucie; Brož, Václav; Arefin, B.; Maaroufi, H. O.; Hurychová, J.; Strnad, Hynek; Žurovec, Michal; Theopold, U.

2016-01-01

Roč. 8, č. 2 (2016), s. 199-210 ISSN 1662-811X R&D Projects: GA ČR GA14-27816S Institutional support: RVO:60077344 ; RVO:68378050 Keywords : chitinase-like proteins * imaginal disc growth factor * hemolymph clot Subject RIV: CE - Biochemistry; EB - Genetics ; Molecular Biology (UMG-J) Impact factor: 3.938, year: 2016 http://www.karger.com/Article/FullText/442351

Distribution and Metabolism of Bt-Cry1Ac Toxin in Tissues and Organs of the Cotton Bollworm, Helicoverpa armigera

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Zhuoya Zhao

2016-07-01

Full Text Available Crystal (Cry proteins derived from Bacillus thuringiensis (Bt have been widely used in transgenic crops due to their toxicity against insect pests. However, the distribution and metabolism of these toxins in insect tissues and organs have remained obscure because the target insects do not ingest much toxin. In this study, several Cry1Ac-resistant strains of Helicoverpa armigera, fed artificial diets containing high doses of Cry1Ac toxin, were used to investigate the distribution and metabolism of Cry1Ac in their bodies. Cry1Ac was only detected in larvae, not in pupae or adults. Also, Cry1Ac passed through the midgut into other tissues, such as the hemolymph and fat body, but did not reach the larval integument. Metabolic tests revealed that Cry1Ac degraded most rapidly in the fat body, followed by the hemolymph, peritrophic membrane and its contents. The toxin was metabolized slowly in the midgut, but was degraded in all locations within 48 h. These findings will improve understanding of the functional mechanism of Bt toxins in target insects and the biotransfer and the bioaccumulation of Bt toxins in arthropod food webs in the Bt crop ecosystem.

Characterization of cyclomorphic stages in the marine tardigrade Halobiotus crispae

DEFF Research Database (Denmark)

Halberg, Kenneth Agerlin; Mortensen, Hans Ramløv; Westh, Peter

2008-01-01

and pseudosimplex 1 (P1) stage. The active stage tolerates large shifts in external salinity. Total body volume of single specimens (350-500 µm) was estimated from microscopical images following salinity transfers from 20 ppt (control) to 2 ppt, 10 ppt and 40 ppt. Our results show that animals in this stage...... experience large changes in total body volume, yet exhibit a regulatory volume decrease/increase over a 48 h period. Hemolymph osmolality was measured by melting point depression in a nanoliter osmometer. In animals kept at 20 ppt the hemolymph osmotic pressure was 926 ± 29 mOsm/kg (n = 6). This value...... changed to 330 ± 50 mOsm/kg (n = 6), 584 ± 68 mOsm/kg (n = 6) and 1297 ± 32 mOsm/kg (n = 6) during exposure to 2 ppt, 10 ppt and 40 ppt respectively. At any given external salinity the active stage hyper-regulates, indicating the excretion of dilute urine. Animals in the P1 stage are freeze tolerant...

Differential infectivity of two Pseudomonas species and the immune response in the milkweed bug, Oncopeltus fasciatus (Insecta: Hemiptera).

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Schneider, M; Dorn, A

2001-10-01

Pseudomonas aeruginosa and Pseudomonas putida show a profound differential infectivity after inoculation in Oncopeltus fasciatus. Whereas P. putida has no significant impact on nymphs, P. aeruginosa kills all experimental animals within 48 h. Both Pseudomonas species, however, induce the same four hemolymph peptides in O. fasciatus. Also injection of saline solution and injury induced these peptides. In general peptide induction was stronger in nymphs than in adult males. A significantly higher number of nymphs survived a challenge with P. aeruginosa when an immunization with P. putida preceded. The antibacterial properties of the hemolymph were demonstrated in inhibition experiments with P. putida. Two of the four inducible peptides (peptides 1 and 4) could be partially sequenced after Edman degradation and were compared with known antibacterial peptides. Peptide 1, of 15 kDa, showed 47.1% identity with the glycine-rich hemiptericin of Pyrrhocoris apterus. Peptide 4, of 2 kDa, had a 77.8% identity with the proline-rich pyrrhocoricin of P. apterus and a 76.9% identity with metalnikowin 1 of Palomena prasina. Peptides 2 and 3 are also small, with molecular weights of 8 and 5 kDa.

Escherichia coli K-12 pathogenicity in the pea aphid, Acyrthosiphon pisum, reveals reduced antibacterial defense in aphids.

Science.gov (United States)

Altincicek, Boran; Ter Braak, Bas; Laughton, Alice M; Udekwu, Klas I; Gerardo, Nicole M

2011-10-01

To better understand the molecular basis underlying aphid immune tolerance to beneficial bacteria and immune defense to pathogenic bacteria, we characterized how the pea aphid Acyrthosiphon pisum responds to Escherichia coli K-12 infections. E. coli bacteria, usually cleared in the hemolymph of other insect species, were capable of growing exponentially and killing aphids within a few days. Red fluorescence protein expressing E. coli K-12 laboratory strain multiplied in the aphid hemolymph as well as in the digestive tract, resulting in death of infected aphids. Selected gene deletion mutants of the E. coli K-12 predicted to have reduced virulence during systemic infections showed no difference in either replication or killing rate when compared to the wild type E. coli strain. Of note, however, the XL1-Blue E. coli K-12 strain exhibited a significant lag phase before multiplying and killing aphids. This bacterial strain has recently been shown to be more sensitive to oxidative stress than other E. coli K-12 strains, revealing a potential role for reactive oxygen species-mediated defenses in the otherwise reduced aphid immune system. Copyright © 2011 Elsevier Ltd. All rights reserved.

Biochemical profile of Achatina fulica (Mollusca: Gastropoda) after infection by different parasitic loads of Angiostrongylus cantonensis (Nematoda, Metastrongylidae).

Science.gov (United States)

Tunholi-Alves, Vinícius Menezes; Tunholi, Victor Menezes; Amaral, Ludimila Santos; Mota, Esther Maria; Maldonado Júnior, Arnaldo; Pinheiro, Jairo; Garcia, Juberlan

2015-01-01

The effect of experimental infection by different parasitic loads of Angiostrongylus cantonensis (Nematode, Metastrongylidae) on the activities of the aminotransferases and concentration of total proteins, uric acid and urea in the hemolymph of Achatina fulica (Mollusca, Gastropoda) were investigated. There was a significant decrease in the concentration of total proteins in the exposed snails to 5000 or more larvae. This change was accompanied by an increase in the concentrations of urea and uric acid in the hemolymph, suggesting a higher rate of deamination of the amino acids. Besides this, variations in the activities of the aminotransferases were also observed, with the highest values recorded in the groups exposed to greater parasite load. These results suggest an increase in the use of total proteins, since there was increased formation of nitrogenous catabolites, in conformity with an increase in the aminotransferase activities. Infection was verified by the fact that L3 larvae recovered from the snails was proportion to the exposure dose of L1 larvae. Histopathological results also indicated presence of an inflammatory cell infiltrate, favoring an increase of both transaminases. Copyright © 2014 Elsevier Inc. All rights reserved.

Anti-Legionella dumoffii Activity of Galleria mellonella Defensin and Apolipophorin III

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Małgorzata Cytryńska

2012-12-01

Full Text Available The gram-negative bacterium Legionella dumoffii is, beside Legionella pneumophila, an etiological agent of Legionnaires’ disease, an atypical form of pneumonia. The aim of this study was to determine the antimicrobial activity of Galleria mellonella defense polypeptides against L. dumoffii. The extract of immune hemolymph, containing a mixture of defense peptides and proteins, exhibited a dose-dependent bactericidal effect on L. dumoffii. The bacterium appeared sensitive to a main component of the hemolymph extract, apolipophorin III, as well as to a defense peptide, Galleria defensin, used at the concentrations 0.4 mg/mL and 40 μg/mL, respectively. L. dumoffii cells cultured in the presence of choline were more susceptible to both defense factors analyzed. A transmission electron microscopy study of bacterial cells demonstrated that Galleria defensin and apolipophorin III induced irreversible cell wall damage and strong intracellular alterations, i.e., increased vacuolization, cytoplasm condensation and the appearance of electron-white spaces in electron micrographs. Our findings suggest that insects, such as G. mellonella, with their great diversity of antimicrobial factors, can serve as a rich source of compounds for the testing of Legionella susceptibility to defense-related peptides and proteins.

Magnetic resonance imaging of live freshwater mussels (Unionidae)

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Michael, Holliman F.; Davis, Denise; Bogan, Arthur E.; Kwak, Thomas J.; Cope, W. Gregory; Levine, Jay F.

2008-01-01

We examined the soft tissues of live freshwater mussels, Eastern elliptio Elliptio complanata, via magnetic resonance imaging (MRI), acquiring data with a widely available human whole-body MRI system. Anatomical features depicted in the profile images included the foot, stomach, intestine, anterior and posterior adductor muscles, and pericardial cavity. Noteworthy observations on soft tissue morphology included a concentration of lipids at the most posterior aspect of the foot, the presence of hemolymph-filled fissures in the posterior adductor muscle, the presence of a relatively large hemolymph-filled sinus adjacent to the posterior adductor muscle (at the ventral-anterior aspect), and segmentation of the intestine (a diagnostic description not reported previously in Unionidae). Relatively little is known about the basic biology and ecological physiology of freshwater mussels. Traditional approaches for studying anatomy and tissue processes, and for measuring sub-lethal physiological stress, are destructive or invasive. Our study, the first to evaluate freshwater mussel soft tissues by MRI, clarifies the body plan of unionid mussels and demonstrates the efficacy of this technology for in vivoevaluation of the structure, function, and integrity of mussel soft tissues.

Evaluation of the toxic effect of endocrine disruptor Bisphenol A (BPA) in the acute and chronic toxicity tests with Pomacea lineata gastropod.

Science.gov (United States)

de Andrade, André Lucas Correa; Soares, Priscila Rafaela Leão; da Silva, Stephannie Caroline Barros Lucas; da Silva, Marília Cordeiro Galvão; Santos, Thamiris Pinheiro; Cadena, Marilia Ribeiro Sales; Soares, Pierre Castro; Cadena, Pabyton Gonçalves

2017-07-01

Bisphenol A (BPA) is a plasticizer and a risk when it interacts with organisms, and can cause changes in the development and reproduction of them. This study aimed to evaluate the effects of BPA, by acute and chronic toxicity tests with neonates and adults of Pomacea lineata. Adults and neonates were divided into groups exposed to BPA (1-20mg/L), or 17β-estradiol (1mg/L) and control in the acute and chronic toxicity tests. Behavior, heart rate, reproduction and hemolymph biochemical analysis were measured. In the acute toxicity test, the 96-h LC 50 with adults was 11.09 and with neonates was 3.14mg/L. In this test, it was observed lethargic behavior and an increase of 77.6% of aspartate aminotransferase in the adults' hemolymph (ptest, it was observed behaviors associated with reproduction, as Copulate, in the groups exposed to BPA. The results that were found in this study proved that BPA is a potentially toxic agent to Pomacea lineata according to biological parameters evaluated. These data contribute to the understanding of BPA toxic effects' in the aquatic invertebrates. Copyright © 2017 Elsevier Inc. All rights reserved.

Behavior of isolated nuclei and chromosomes of Rhynchosciara in vitro

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Cestari, A N; Simoes, L C.G. [Sao Paulo Univ. (Brazil). Inst. de Biociencias

1980-09-01

Nuclei and chromosomes were isolated from salivary glands of Rhynchosciara by a method involving micropipette manipulation, after diluting the intercellular cement with a mixture of citric acid, tween 80 and sucrose. Culture media supplemented and not supplemented with different concentrations of lobster hemolymph or calf serum, were compared. In the best medium, isolated nuclei and chromosomes treated with tritiated precursors showed nucleic acid and protein synthesis.

Effects of chronic waterborne nickel exposure on growth, ion homeostasis, acid-base balance, and nickel uptake in the freshwater pulmonate snail, Lymnaea stagnalis.

Science.gov (United States)

Niyogi, Som; Brix, Kevin V; Grosell, Martin

2014-05-01

The freshwater pulmonate snail, Lymnaea stagnalis, is the most sensitive aquatic organism tested to date for Ni. We undertook a series of experiments to investigate the underlying mechanism(s) for this observed hypersensitivity. Consistent with previous experiments, juvenile snail growth in a 21-day exposure was reduced by 48% relative to the control when exposed to 1.3 μg l(-1) Ni (EC20 less than the lowest concentration tested). Ca(2+) homeostasis was significantly disrupted by Ni exposure as demonstrated by reductions in net Ca(2+) uptake, and reductions in Ca(2+) concentrations in the hemolymph and soft tissues. We also observed reduced soft tissue [Mg(2+)]. Snails underwent a significant alkalosis with hemolymph pH increasing from 8.1 to 8.3 and hemolymph TCO2 increasing from 19 to 22 mM in control versus Ni-exposed snails, respectively. Unlike in previous studies with Co and Pb, snail feeding rates were found to be unaffected by Ni at the end of the exposure. Snails accumulated Ni in the soft tissue in a concentration-dependent manner, and Ni uptake experiments with (63)Ni revealed a biphasic uptake profile - a saturable high affinity component at low exposure concentrations (36-189 nM) and a linear component at the high exposure concentrations (189-1,897 nM). The high affinity transport system had an apparent Km of 89 nM Ni(2+) and Vmax of 2.4 nmol g(-1)h(-1). This equates to a logK of 7.1, significantly higher than logK's (2.6-5.2) for any other aquatic organisms evaluated to date, which will have implications for Biotic Ligand Model development. Finally, pharmacological inhibitors that block Ca(2+) uptake pathways in snails did not inhibit Ni uptake, suggesting that the uptake of Ni does not occur via Ca(2+) uptake pathways. As with Cu and Pb, the exact mechanism for the significant disruption in Ca(2+) homeostasis and reduction in juvenile snail growth remains unknown. Copyright © 2014 Elsevier B.V. All rights reserved.

Changes in energetic metabolism of Biomphalaria glabrata (Mollusca, Planorbidae in response to exogenous calcium

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L. D. Silva

Full Text Available Abstract Calcium is considered an essential element for the metabolism of aquatic snail Biomphalaria glabrata (Say, 1818, intermediate host of Schistosoma mansoni Sambon, 1907 in Brazil, and represents a limiting factor to its distribution and adaptation to the environment. This study investigated the effect of different concentrations of exogenous CaCO3 on the energetic metabolism of B. glabrata for better understanding the physiological interference of chemical elements dissolved in the environment with the physiology of this species. Sixty-day-old snails were distributed into six groups, five exposed to different concentrations of CaCO3 (20, 40, 60, 80 and 100 mg/L and a control group. The exposure to CaCO3 was assessed over time, with analysis of 15 snails of each group in the following intervals: 1, 14, 21 or 30 days for hemolymph extraction. Concentrations of calcium and glucose in the hemolymph were determined by commercial kits, and organic acids were extracted using an ion exchange column and analyzed by high-performance liquid chromatography. Concentration of calcium in the hemolymph showed no significant difference (p>0.05 from the control group and between the concentrations tested. Concentration of glucose decreased (p<0.05 in the treatments of exposure to 20 and 40 mg/L and increased when exposed to 80 and 100 mg/L CaCO3 compared to control and to other concentrations tested over 30 days. The organic acids pyruvate, oxaloacetate, citrate, succinate, fumarate, beta-hydroxybutyrate and lactate presented increased concentrations, while propionate and acetoacetate, decreased concentrations, when exposed to CaCO3 compared to control. Considering the influence of different periods of exposure to CaCO3, on the 14th day, there were stronger alterations in the metabolism of B. glabrata. In conclusion, exposure to CaCO3 reduced the concentration of glucose, which is metabolized into pyruvate, the final product of glycolysis, and also

Ethnomedicines used in Trinidad and Tobago for urinary problems and diabetes mellitus

Science.gov (United States)

Lans, Cheryl A

2006-01-01

Background This paper is based on ethnobotanical interviews conducted from 1996–2000 in Trinidad and Tobago with thirty male and female respondents. Methods A non-experimental validation was conducted on the plants used for urinary problems and diabetes mellitus: This is a preliminary step to establish that the plants used are safe or effective, to help direct clinical trials, and to inform Caribbean physicians of the plants' known properties to avoid counter-prescribing. Results The following plants are used to treat diabetes: Antigonon leptopus, Bidens alba, Bidens pilosa, Bixa orellana, Bontia daphnoides, Carica papaya, Catharanthus roseus, Cocos nucifera, Gomphrena globosa, Laportea aestuans, Momordica charantia, Morus alba, Phyllanthus urinaria and Spiranthes acaulis. Apium graviolens is used as a heart tonic and for low blood pressure. Bixa orellana, Bontia daphnoides, Cuscuta americana and Gomphrena globosa are used for jaundice. The following plants are used for hypertension: Aloe vera, Annona muricata, Artocarpus altilis, Bixa orellana, Bidens alba, Bidens pilosa, Bonta daphnoides, Carica papaya, Cecropia peltata, Citrus paradisi, Cola nitida, Crescentia cujete, Gomphrena globosa, Hibiscus sabdariffa, Kalanchoe pinnata, Morus alba, Nopalea cochinellifera, Ocimum campechianum, Passiflora quadrangularis, Persea americana and Tamarindus indicus. The plants used for kidney problems are Theobroma cacao, Chamaesyce hirta, Flemingia strobilifera, Peperomia rotundifolia, Petiveria alliacea, Nopalea cochinellifera, Apium graveolens, Cynodon dactylon, Eleusine indica, Gomphrena globosa, Pityrogramma calomelanos and Vetiveria zizanioides. Plants are also used for gall stones and for cooling. Conclusion Chamaesyce hirta, Cissus verticillata, Kalanchoe pinnata, Peperomia spp., Portulaca oleraceae, Scoparia dulcis, and Zea mays have sufficient evidence to support their traditional use for urinary problems, "cooling" and high cholesterol. Eggplant extract as a

Ethnomedicines used in Trinidad and Tobago for urinary problems and diabetes mellitus

Directory of Open Access Journals (Sweden)

Lans Cheryl A

2006-10-01

Full Text Available Abstract Background This paper is based on ethnobotanical interviews conducted from 1996–2000 in Trinidad and Tobago with thirty male and female respondents. Methods A non-experimental validation was conducted on the plants used for urinary problems and diabetes mellitus: This is a preliminary step to establish that the plants used are safe or effective, to help direct clinical trials, and to inform Caribbean physicians of the plants' known properties to avoid counter-prescribing. Results The following plants are used to treat diabetes: Antigonon leptopus, Bidens alba, Bidens pilosa, Bixa orellana, Bontia daphnoides, Carica papaya, Catharanthus roseus, Cocos nucifera, Gomphrena globosa, Laportea aestuans, Momordica charantia, Morus alba, Phyllanthus urinaria and Spiranthes acaulis. Apium graviolens is used as a heart tonic and for low blood pressure. Bixa orellana, Bontia daphnoides, Cuscuta americana and Gomphrena globosa are used for jaundice. The following plants are used for hypertension: Aloe vera, Annona muricata, Artocarpus altilis, Bixa orellana, Bidens alba, Bidens pilosa, Bonta daphnoides, Carica papaya, Cecropia peltata, Citrus paradisi, Cola nitida, Crescentia cujete, Gomphrena globosa, Hibiscus sabdariffa, Kalanchoe pinnata, Morus alba, Nopalea cochinellifera, Ocimum campechianum, Passiflora quadrangularis, Persea americana and Tamarindus indicus. The plants used for kidney problems are Theobroma cacao, Chamaesyce hirta, Flemingia strobilifera, Peperomia rotundifolia, Petiveria alliacea, Nopalea cochinellifera, Apium graveolens, Cynodon dactylon, Eleusine indica, Gomphrena globosa, Pityrogramma calomelanos and Vetiveria zizanioides. Plants are also used for gall stones and for cooling. Conclusion Chamaesyce hirta, Cissus verticillata, Kalanchoe pinnata, Peperomia spp., Portulaca oleraceae, Scoparia dulcis, and Zea mays have sufficient evidence to support their traditional use for urinary problems, "cooling" and high cholesterol

Behavior of isolated nuclei and chromosomes of Rhynchosciara in vitro

International Nuclear Information System (INIS)

Cestari, A.N.; Simoes, L.C.G.

1980-01-01

Nuclei and chromosomes were isolated from salivary glands of Rhynchosciara by a method involving micropipette manipulation, after diluting the intercellular cement with a mixture of citric acid, tween 80 and sucrose. Culture media supplemented and not supplemented with different concentrations of lobster hemolymph or calf serum, were compared. In the best medium, isolated nuclei and chromosomes treated with tritiated precursors showed nucleic acid and protein synthesis. (Author) [pt

Pre-treatment with melatonin decreases abamectin induced toxicity in a nocturnal insect Spodoptera litura (Lepidoptera: Noctuidae).

Science.gov (United States)

Subala, Subramanian P; Zubero, Eduardo E; Alatorre-Jimenez, Moises A; Shivakumar, Muthugounder S

2017-12-01

Oxidative stress is an important component of the mechanism of pesticide toxicity. The aim of the present study was to investigate the time-dependent melatonin effects against abamectin-induced oxidative stress in a S.litura model. Larvae were divided into 5 different groups; (1) control group,(2) Melatonin group (4.3×10 -5 M/100ml diet), (3) Abamectin group 1.5ml/L, (4) Pre-melatonin treated group (PM) (4.3×10 -5 M/100ml diet) before abamectin exposure 1.5ml/L, (5) Post-melatonin treated group (TM) after abamectin exposure. Melatonin was supplemented via artificial diet in PM and TM animals during 24h. Midgut, fatbody, and hemolymph, were collected for the analysis of oxidative stress markers (Total ROS, GSH, nitrite, TBARS, LPO), antioxidant enzyme levels (SOD, GST, CAT, POX, APOX) in fifth instar larvae. Midgut damage was examined by using morphological analysis. Our results observed that ABA group showed significant changes (pmelatonin. Significant (pmelatonin treatment reduces this damage due to its antioxidant properties, especially POX levels in midgut, fatbody, and hemolymph. Therefore, indoleamine can play a vital role curtailing the abamectin toxicity in time dependent manner in S.litura. Copyright © 2017 Elsevier B.V. All rights reserved.

Parasitoid-host endocrine relations: self-reliance or co-optation?

Science.gov (United States)

Cole, T J; Beckage, N E; Tan, F F; Srinivasan, A; Ramaswamy, S B

2002-12-01

High titers of juvenile hormone (JH) maintain developmental arrest in Manduca sexta larvae parasitized by Cotesia congregata. Parasitized hosts exhibit up to 9.5 times greater amounts of total hemolymph JH (from 0.6+/-0.09 to 2.51+/-0.43ng/ml) compared to non-parasitized controls. Elevated titers are observed throughout the fifth instar, even beyond egression of the parasitoids on day 5. GC-MS analysis revealed that in hemolymph of unparasitized control larvae, JH I is the major homolog and levels of JH III are negligible; in parasitized individuals the amounts of JH I, II, and III rise, and JH III predominates. Neck ligation ensured separation of M. sexta's corpora allata from the posterior section, which contained most of the parasitoids in the infected insects. When the posterior region was sampled, JHs were not detected in the non-parasitzed larvae, but in those parasitized, JH III was found (1.98+/-0.29ng/ml, 24 h post-ligation). JH III was the only homolog produced and secreted by the parasitoid in in vitro culture. This is the first report stating that a parasitoid secretes JH III and may contribute, at least in part, to the circulating titer in the host hemocoel, concurrently promoting host production of JH I and II.

Influence of periodic heartbeat reversal and abdominal movements on hemocoelic and tracheal pressure in resting blowflies Calliphora vicina.

Science.gov (United States)

Wasserthal, Lutz Thilo

2012-01-15

In Calliphoridae and Drosophilidae, the dorsal vessel (heart and aorta with associated venous channels) is the only connection between the thorax and the abdomen. Hemolymph oscillates between the compartments by periodic heartbeat reversal, but both the mechanism and its influence on hemocoelic and tracheal pressure have remained unclear. The pumping direction of the heart regularly reverses, with a higher pulse rate during backward compared with forward pumping. A sequence of forward and backward pulse periods lasts approximately 34 s. Pulse rate, direction, velocity and the duration of heartbeat periods were determined by thermistor and electrophysiological measurements. For the first time, heartbeat-induced pressure changes were measured in the hemocoel and in the tracheal system of the thorax and the abdomen. The tracheal pressure changed from sub-atmospheric during backward heartbeat to supra-atmospheric during forward heartbeat in the thorax and inversely in the abdomen. The heartbeat reversals were coordinated with slow abdominal movements with a pumping stroke at the beginning of the forward pulse period. The pressure effect of the pumping stroke was visible only in the abdomen. Periodic hemolymph shift and abdominal movements resulted in pressure changes in the hemocoel and tracheal system alternating in the thorax and abdomen, suggesting an effect on respiratory gas exchange.

Host and environmental factors influencing "Candidatus Liberibacter asiaticus" acquisition in Diaphorina citri.

Science.gov (United States)

Wu, Fengnian; Huang, Jiaquan; Xu, Meirong; Fox, Eduardo G P; Beattie, G Andrew C; Holford, Paul; Cen, Yijing; Deng, Xiaoling

2018-05-03

Diaphorina citri is a vector of "Candidatus Liberibacter asiaticus" (CLas) associated with citrus Huanglongbing. In this study, the infection and titers of CLas in the psyllid, were monitored for life cycle stage, sex, host-plant CLas titer, host-plant genotype, and ambient temperature. Acquisition efficiency of CLas by D. citri was highest in nymphs reared at 25 °C on a host plant with high CLas titers but was independent of the host genotypes assessed and of vector sex. We further observed that D. citri nymphs acquired CLas more rapidly than adults based on acquisition access periods (AAPs). CLas did not multiply in the alimentary canal, hemolymph, and salivary glands of adults for 18 d after a 3-day AAP as adult. However, CLas multiplication was detected in hemolymph and salivary gland of adults after the bacterium was acquired by nymphs. Eighty percent of salivary glands of adults contained CLas 18 d after a 3-day AAP as nymph compared to 10% 18 d after a 3-day AAP as adults. Different factors tested herein influenced CLas acquisition efficiency of D. citri, CLas multiplication and spread inside the psyllid. These observations serve to better understand mechanisms of CLas infection in D. citri. This article is protected by copyright. All rights reserved.

cis-Canthaxanthins: unusual carotenoids in the eggs and the reproductive system of female brine shrimp Artemia

OpenAIRE

Nelis, H.J.C.F.; Lavens, P.; Moens, L.; Sorgeloos, P.; Jonckheere, J.A.; Criel, G.R.J.; De Leenheer, A.P.

1984-01-01

The significance of carotenoid accumulation in crustacean eggs remains obscure, particularly because neither eggs nor female animals have been found to display specific pigment patterns in relation to reproduction. We report here the first example of carotenoids found exclusively in the ovaries, the eggs, and the hemolymph, but not in the carcass of a female, reproductively active crustacean, i.e. the brine shrimp Artemia . These pigments are virtually absent in males and in immature animals ...

Breaking good: a chemist wanders into entomology.

Science.gov (United States)

Law, John H

2015-01-07

In this highly personal account of my career in science, I try to show how many others influenced its course. I was able to abandon work in pure chemistry and microbiology and to take up research in entomology only with the help of others. My faith in the value of collaborative, interdisciplinary work has been the key to success. Our focus on proteins of insect hemolymph has provided valuable insights into insect biochemistry and physiology.

Morel-Lavallée lesion: A closed degloving injury that requires real attention

International Nuclear Information System (INIS)

Nair, Anirudh V; Nazar, PK; Sekhar, Resmi; Ramachandran, PV; Moorthy, Srikanth

2014-01-01

Morel-Lavallée lesions are post-traumatic, closed degloving injuries occurring deep to subcutaneous plane due to disruption of capillaries resulting in an effusion containing hemolymph and necrotic fat. Magnetic resonance imaging (MRI) is the modality of choice in the evaluation of Morel-Lavallée lesion. Early diagnosis and management is essential as any delay in diagnosis or missed lesion will lead to the effusion becoming infected or leading to extensive skin necrosis

Morel-Lavallée lesion: A closed degloving injury that requires real attention.

Science.gov (United States)

Nair, Anirudh V; Nazar, Pk; Sekhar, Resmi; Ramachandran, Pv; Moorthy, Srikanth

2014-07-01

Morel-Lavallée lesions are post-traumatic, closed degloving injuries occurring deep to subcutaneous plane due to disruption of capillaries resulting in an effusion containing hemolymph and necrotic fat. Magnetic resonance imaging (MRI) is the modality of choice in the evaluation of Morel-Lavallée lesion. Early diagnosis and management is essential as any delay in diagnosis or missed lesion will lead to the effusion becoming infected or leading to extensive skin necrosis.

Fast and Cost-Effective Biochemical Spectrophotometric Analysis of Solution of Insect “Blood” and Body Surface Elution

OpenAIRE

Aleksandra Łoś; Aneta Strachecka

2018-01-01

Using insect hemolymph (“blood”) and insect body surface elutions, researchers can perform rapid and cheap biochemical analyses to determine the insect’s immunology status. The authors of this publication describe a detailed methodology for a quick marking of the concentration of total proteins and evaluation of the proteolytic system activity (acid, neutral, and alkaline proteases and protease inhibitors), as well as a methodology for quick “liver” tests in insects: alanine aminotransferase ...

Trigona corvina: An Ecological Study Based on Unusual Nest Structure and Pollen Analysis

Directory of Open Access Journals (Sweden)

David W. Roubik

2009-01-01

Full Text Available We found that the nest of Trigona corvina (Apidae; Meliponini consists mainly of pollen exines from bee excrement, forming a scutellum shield encasing the colony. A 20-year-old nest (1980–2000 from a lowland Panama forested habitat was sawed in half longitudinally, and a 95 cm transect was systematically sampled each 5 cm. Samples subjected to detailed pollen analysis held 72 botanical species belonging to 65 genera in 41 families. Over 90% of scutellum pollen volume was Cecropiaceae and Arecaceae, among >1013 grains. Potentially the oldest samples, in the middle of the nest, indicate that Mimosoideae, Euphorbiaceae, and Bombacaceae (now Malvaceae were lost when Africanized honey bee competitors colonized Panama in 1984. Cecropia deposited in the nest increased markedly after landscape-level vegetation disturbance. Pollen from Cavanillesia demonstrated that the foraging range encompassed 3 km2 and perhaps 500 plant species. Trigona corvina primarily foraged on plants with large inflorescences, consistent with foraging theory considering their aggressive behavior.

Oxygen consumption remains stable while ammonia excretion is reduced upon short time exposure to high salinity in Macrobrachium acanthurus (Caridae: Palaemonidae, a recent freshwater colonizer

Directory of Open Access Journals (Sweden)

Carolina A. Freire

2017-10-01

Full Text Available ABSTRACT Palaemonid shrimps occur in the tropical and temperate regions of South America and the Indo-Pacific, in brackish/freshwater habitats, and marine coastal areas. They form a clade that recently (i.e., ~30 mya invaded freshwater, and one included genus, Macrobrachium Bate, 1868, is especially successful in limnic habitats. Adult Macrobrachium acanthurus (Wiegmann, 1836 dwell in coastal freshwaters, have diadromous habit, and need brackish water to develop. Thus, they are widely recognized as euryhaline. Here we test how this species responds to a short-term exposure to increased salinity. We hypothesized that abrupt exposure to high salinity would result in reduced gill ventilation/perfusion and decreased oxygen consumption. Shrimps were subjected to control (0 psu and experimental salinities (10, 20, 30 psu, for four and eight hours (n = 8 in each group. The water in the experimental containers was saturated with oxygen before the beginning of the experiment; aeration was interrupted before placing the shrimp in the experimental container. Dissolved oxygen (DO, ammonia concentration, and pH were measured from the aquaria water, at the start and end of each experiment. After exposure, the shrimp’s hemolymph was sampled for lactate and osmolality assays. Muscle tissue was sampled for hydration content (Muscle Water Content, MWC. Oxygen consumption was not reduced and hemolymph lactate did not increase with increased salinity. The pH of the water decreased with time, under all conditions. Ammonia excretion decreased with increased salinity. Hemolymph osmolality and MWC remained stable at 10 and 20 psu, but osmolality increased (~50% and MWC decreased (~4% at 30 psu. The expected reduction in oxygen consumption was not observed. This shrimp is able to tolerate significant changes in water salt concentrations for a few hours by keeping its metabolism in aerobic mode, and putatively shutting down branchial salt uptake to avoid massive salt

Evaluation of the immune responses of the brown mussel Perna perna as indicators of fecal pollution.

Science.gov (United States)

Silva Dos Santos, Fernanda; Neves, Raquel Almeida Ferrando; Carvalho, Wanderson Fernandes de; Krepsky, Natascha; Crapez, Mirian Araújo Carlos

2018-06-01

The mussel Perna perna is an intertidal bivalve that is widely distributed, cultivated and consumed in South Africa, Brazil and Venezuela. Among marine resources, bivalve mollusks are one of the most impacted by anthropogenic pollution, as they can accumulate pathogenic bacteria and water pollutants. Hemocytes are molluscan defense cells, and their abundance and functions can be affected in response to contaminants, such as bacterial load. However, no previous study has investigated the immune response of P. perna hemocytes. The aim of this study was to evaluate several immune parameters in P. perna as indicators of fecal pollution in mussel hemolymph and in seawater. We collected mussels and adjacent seawater from beaches with different levels of fecal contamination in Rio de Janeiro state (Brazil): Vermelha Beach (VB); Icaraí Beach (IB); Urca Beach (UB); and Jurujuba Beach (JB). Hemocyte parameters (density, morphology, phagocytic activity and production of Reactive Oxygen Species - ROS) were evaluated using flow cytometry. We quantified Fecal Indicator Bacteria (FIB) in seawater by the multiple tubes technique for each beach and for hemolymph by the spread-plate technique. In agreement with historical evaluation of fecal contamination levels, UB presented the highest FIB abundance in seawater (thermotolerant coliforms, TEC = 1600 NMP 100 mL -1 ), whereas VB exhibited the lowest (TEC = 17 NMP 100 mL -1 ). UB mussels had six and eight times higher hemocyte density and phagocytic activity, respectively, than mussels from VB. Mussels from VB and IB presented a significantly lower number of total coliforms in hemolymph and a significantly higher relative internal complexity of hemocytes than those from UB and JB (p ≤ 0.01, PERMANOVA). ROS production by hemocytes was significantly lower in mussels from VB compared to those from JB (p = 0.04, ANOVA). Our results indicate a significant relationship between the level of fecal contamination in

The ultrastructure of book lung development in the bark scorpion Centruroides gracilis (Scorpiones: Buthidae

Directory of Open Access Journals (Sweden)

Farley Roger D

2011-07-01

Full Text Available Abstract Background Near the end of the nineteenth century the hypothesis was presented for the homology of book lungs in arachnids and book gills in the horseshoe crab. Early studies with the light microscope showed that book gill lamellae are formed by outgrowth and possibly some invagination (infolding of hypodermis (epithelium from the posterior surface of opisthosomal limb buds. Scorpion book lungs are formed near the bilateral sites of earlier limb buds. Hypodermal invaginations in the ventral opisthosoma result in spiracles and sac-like cavities (atria. In early histological sections of embryo book lungs, widening of the atrial entrance of some lamellae (air channels, air sacs, saccules was interpreted as an indication of invagination as hypothesized for book gill lamellae. The hypodermal infolding was thought to produce the many rows of lamellar precursor cells anterior to the atrium. The ultrastructure of scorpion book lung development is compared herein with earlier investigations of book gill formation. Results In scorpion embryos, there is ingression (inward migration of atrial hypodermal cells rather than invagination or infolding of the atrial hypodermal layer. The ingressing cells proliferate and align in rows anterior to the atrium. Their apical-basal polarity results in primordial air channels among double rows of cells. The cuticular walls of the air channels are produced by secretion from the apical surfaces of the aligned cells. Since the precursor cells are in rows, their secreted product is also in rows (i.e., primordial air channels, saccules. For each double row of cells, their opposed basal surfaces are gradually separated by a hemolymph channel of increasing width. Conclusions The results from this and earlier studies show there are differences and similarities in the formation of book lung and book gill lamellae. The homology hypothesis for these respiratory organs is thus supported or not supported depending on which

[Effect of population density on enzymatic activity of antioxidative and phenol oxidase systems of imagoes and nymphs of the marble cockroach Nauphoeta cinerea].

Science.gov (United States)

Murzagulov, G S; Saltykova, E S; Gaĭfullina, L R; Nikolenko, A G

2013-01-01

The work deals with effect of density of population on functional activity of components pf protective system of adult individuals and nymphs of the marble cockroach. The resistance of individuals has been noted to decrease both at individual maintenance and under conditions of overpopulation. Changes in activities of enzymes of antioxidative and phenoloxidase systems are studied ion the insect hemolymph and intestine. Possible consequences of isolation and overpopulation are discussed both for stability and for individual development.

Sequestration of Glucosinolates and Iridoid Glucosides in Sawfly Species of the Genus Athalia and Their Role in Defense Against Ants

DEFF Research Database (Denmark)

Opitz, Sebastian E. W.; Jensen, Søren Rosendal; Müller, Caroline

2010-01-01

In this study, the larval sequestration abilities and defense effectiveness of four sawfly species of the genus Athalia (Hymenoptera: Tenthredinidae) that feed as larvae either on members of the Brassicaceae or Plantaginaceae were investigated. Brassicaceae are characterized by glucosinolates (GLSs...... hemolymph of the GLSsequestering conspicuous A. rosae larvae. The results show that glucoside sequestration is widespread in the genus Athalia, but that the specific glucoside uptake can result in different defense effectiveness against a predator species....

Viral epidemiology of the adult Apis Mellifera infested by the Varroa destructor mite

OpenAIRE

Bernardi, Sara; Venturino, Ezio

2016-01-01

The ectoparasitic mite Varroa destructor has become one of the major worldwide threats for apiculture. Varroa destructor attacks the honey bee Apis mellifera weakening its host by sucking hemolymph. However, the damage to bee colonies is not strictly related to the parasitic action of the mite but it derives, above all, from its action as vector increasing the transmission of many viral diseases such as acute paralysis (ABPV) and deformed wing viruses (DWV), that are considered among the main...

The U2 snDNA Is a Useful Marker for B Chromosome Detection and Frequency Estimation in the Grasshopper Abracris flavolineata.

Science.gov (United States)

Milani, Diogo; Palacios-Gimenez, Octavio M; Cabral-de-Mello, Diogo C

2017-01-01

In this study, we describe a strategy to determine the presence of B chromosomes in the living grasshopper Abracris flavolineata by FISH using U2 snDNA as a probe in interphase hemolymph nuclei. In individuals without B chromosomes, (0B) 2 dot signals were noticed, corresponding to A complement U2 snDNA clusters. In +1B and +2B individuals, 4 or 8 additional signals were noticed, respectively. In all cases, the absence or presence of 1 or 2 B chromosomes correlated in hemolymph and in somatic or germline tissues, validating the efficiency of the marker. Our data suggest that the B chromosome of A. flavolineata is present in all somatic tissues. B-carrying individuals showed the same number of B chromosomes in germ and somatic cells, suggesting that the B is mitotically stable. The marker was used to compare B chromosome frequency in the analyzed population with a sample collected previously, in order to test for B frequency changes and differences of B chromosome prevalence among sexes, but no statistically significant differences were noticed. The identification of living animals harboring B chromosomes will be very useful in future studies of B chromosome transmission, as well as in functional studies involving RNA analysis, thus contributing to the understanding of evolutionary history and the possible role of the B chromosome in A. flavolineata. © 2017 S. Karger AG, Basel.

Dynamics of vitellogenin and vitellogenesis-inhibiting hormone levels in adult and subadult whiteleg shrimp, Litopenaeus vannamei: relation to molting and eyestalk ablation.

Science.gov (United States)

Kang, Bong Jung; Okutsu, Tomoyuki; Tsutsui, Naoaki; Shinji, Junpei; Bae, Sun-Hye; Wilder, Marcy N

2014-01-01

Levels of vitellogenin (VG) and vitellogenesis-inhibiting hormone (VIH) in the whiteleg shrimp, Litopenaeus vannamei, were measured by time-resolved fluoroimmunoassay in relation to the molting cycle and ovarian maturation induced by eyestalk ablation. During the molt cycle, VG mRNA expression levels and VG concentrations showed similar patterns of fluctuation. VG levels increased significantly at early intermolt (stage C0) in adults, but not in subadults. Unilateral and bilateral eyestalk ablation increased VG levels in adults, whereas only bilateral eyestalk ablation affected subadults. VIH levels showed contrasting patterns between adults and subadults. In adults, levels were high in late postmolt adults (stage B) and then low thereafter, whereas they increased from postmolt (stage A) to intermolt (stage C0) in subadults and remained high. Unilateral eyestalk ablation increased VIH levels 10 days following ablation in adults, after which levels decreased at 20 days. VIH levels decreased from 10 to 20 days after bilateral ablation. Both unilateral and bilateral ablation led to increased VIH levels in subadults. Eyestalk ablation induced ovarian maturation, but did not reduce VIH concentrations in the hemolymph. This phenomenon was perhaps due to other crustacean hyperglycemic hormone peptides having cross-reactivity with VIH antibodies. This is the first report to quantify concentrations of VG and VIH together in L. vannamei hemolymph, and to examine their relative dynamics.

Hemocyte-mediated phagocytosis differs between honey bee (Apis mellifera) worker castes.

Science.gov (United States)

Hystad, Eva Marit; Salmela, Heli; Amdam, Gro Vang; Münch, Daniel

2017-01-01

Honey bees as other insects rely on the innate immune system for protection against diseases. The innate immune system includes the circulating hemocytes (immune cells) that clear pathogens from hemolymph (blood) by phagocytosis, nodulation or encapsulation. Honey bee hemocyte numbers have been linked to hemolymph levels of vitellogenin. Vitellogenin is a multifunctional protein with immune-supportive functions identified in a range of species, including the honey bee. Hemocyte numbers can increase via mitosis, and this recruitment process can be important for immune system function and maintenance. Here, we tested if hemocyte mediated phagocytosis differs among the physiologically different honey bee worker castes (nurses, foragers and winter bees), and study possible interactions with vitellogenin and hemocyte recruitment. To this end, we adapted phagocytosis assays, which-together with confocal microscopy and flow cytometry-allow qualitative and quantitative assessment of hemocyte performance. We found that nurses are more efficient in phagocytic uptake than both foragers and winter bees. We detected vitellogenin within the hemocytes, and found that winter bees have the highest numbers of vitellogenin-positive hemocytes. Connections between phagocytosis, hemocyte-vitellogenin and mitosis were worker caste dependent. Our results demonstrate that the phagocytic performance of immune cells differs significantly between honey bee worker castes, and support increased immune competence in nurses as compared to forager bees. Our data, moreover, provides support for roles of vitellogenin in hemocyte activity.

Heterologous expression, purification and characterization of human β-1,2-N-acetylglucosaminyltransferase II using a silkworm-based Bombyx mori nucleopolyhedrovirus bacmid expression system.

Science.gov (United States)

Miyazaki, Takatsugu; Kato, Tatsuya; Park, Enoch Y

2018-02-03

β-1,2-N-Acetylglucosaminyltransferase II (GnTII, EC 2.4.1.143) is a Golgi-localized type II transmembrane enzyme that catalyzes the transfer of N-acetylglucosamine to the 6-arm of the trimanosyl core of N-glycans, an essential step in the conversion of oligomannose-type to complex-type N-glycans. Despite its physiological importance, there have been only a few reports on the heterologous expression and structure-function relationship of this enzyme. Here, we constructed a silkworm-based Bombyx mori nucleopolyhedrovirus bacmid expression system and expressed human GnTII (hGnTII) lacking the N-terminal cytosolic tail and transmembrane region. The recombinant hGnTII was purified from silkworm larval hemolymph in two steps by using tandem affinity purification tags, with a yield of approximately 120 μg from 10 mL hemolymph, and exhibited glycosyltransferase activity and strict substrate specificity. The enzyme was found to be N-glycosylated by the enzymatic cleavage of glycans, while hGnTII expressed in insect cells had not been reported to be glycosylated. Although insects typically produce pauci-mannosidic-type glycans, the structure of N-glycans in the recombinant hGnTII was suggested to be of the complex type, and the removal of the glycans did not affect the enzymatic activity. Copyright © 2018 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

An increase in insulin is important for the acquisition conditioned taste aversion in Lymnaea.

Science.gov (United States)

Mita, Koichi; Yamagishi, Miki; Fujito, Yutaka; Lukowiak, Ken; Ito, Etsuro

2014-12-01

Conditioned taste aversion (CTA) in Lymnaea is brought about by pairing a sucrose solution (the conditioned stimulus, CS) with an electric shock (the unconditioned stimulus, US). Following repeated CS-US pairings, CTA occurs and it is consolidated into long-term memory (LTM). The best CTA is achieved, if snails are food-deprived for 1 day before training commences. With a longer period of food deprivation (5 days), learning and memory formation does not occur. It has been hypothesized that the levels of insulin in the central nervous system (CNS) are very important for CTA to occur. To test his hypothesis, we injected insulin directly into 5-day food-deprived snails. The injection of insulin, as expected, resulted in a decrease in hemolymph glucose concentration. Consistent with our hypothesis with insulin injection, learning and memory formation of CTA occurred. That is, the 'insulin spike' is more important than an increase in hemolymph glucose concentration for CTA-LTM. If we injected an insulin receptor antibody into the snails before the insulin injection, learning was formed but memory formation was not, which is consistent with our previous study. Therefore, a rise in the insulin concentration (i.e., insulin spike) in the CNS is considered to be a key determining factor in the process of CTA-LTM. Copyright © 2014 Elsevier Inc. All rights reserved.

Evaluation of radiosensitivity hemocytes of Biomphalaria glabrata exposed to gamma radiation

International Nuclear Information System (INIS)

Silva, L.R.S.; Amaral, A.J.; Silva, E.B.; Amancio, F.F.; Melo, A.M.M.A.

2013-01-01

The mollusc Biomphalaria glabrata have characteristics that allow them to be identified as an animal model ideal for monitoring areas exposed to chemical agents and physical. This study evaluated the effect of ionizing radiation from Cobalt-60 in haemocytes present in the hemolymph of Biomphalaria glabrata, with the goal of using these cells as indicators of the presence of radiation in aquatic environments. The mollusks were divided into five groups: one control and four subjected doses of 25, 35, 45 and 55 Gy of gamma radiation. After 48 hours of irradiation, the clam hemolymph was collected and slides were prepared and stained with Giemsa for analyses under a light microscope. Statistical analysis was performed using ANOVA and Tukey's test, p <0.05. The results showed that the total number of cells after irradiation reduced compared to control except at a dose of 55 Gy. During data analysis, morphological changes were observed in haemocytes of mollusks subjected to doses of 35, 45 and 55 Gy. These modifications consisted of nucleus bilobulated and nucleo plasmatic bridges. Another change was exclusively observed in the cellular exposure of 55 Gy, where hemocytes showed misshapen nuclei and cytoplasm vacuolisation, suggestive of apoptosis. It is concluded that hemocytes are sensitive to radiation and can be used as indicators of the presence of high doses of ionizing radiation in aquatic environments. (author)

Responses of free radical metabolism to air exposure or salinity stress, in crabs (Callinectes danae and C. ornatus) with different estuarine distributions.

Science.gov (United States)

Freire, Carolina A; Togni, Valéria G; Hermes-Lima, Marcelo

2011-10-01

The swimming crabs Callinectes danae and C. ornatus are found in bays and estuaries, but C. danae is more abundant in lower salinities, while C. ornatus remains restricted to areas of higher salinity. Experimental crabs of both species were submitted to: air exposure (Ae, 3h), reimmersion in 33‰ (control) sea water (SW) (Ri, 1h) following air exposure; hyposaline (Ho, 10‰ for 2h) or hypersaline (He, 40‰ for 2h) SW, then return to control 33‰ SW (RHo and RHe, for 1h). Hemolymph was sampled for osmolality and chloride determinations. Activity of antioxidant enzymes [glutathione peroxidase (GPX), catalase, glutathione-S-transferase] and levels of carbonyl proteins and lipid peroxidation (TBARS) were evaluated in hepatopancreas, muscle, anterior and posterior gills. In Ho groups, hemolymph concentrations were lower in both species, compared to He groups. C. danae displayed higher control activities of GPX (hepatopancreas and muscle) and catalase (all four tissues) than C. ornatus. C. ornatus presented increased activities of catalase and GPX in Ae, Ri, and He groups. Increased TBARS was seen in C. ornatus tissues (He group). The more euryhaline species displayed higher constitutive activities of antioxidant enzymes, and the less euryhaline species exhibited activation of these enzymes when exposed to air or hyper-salinity. Copyright © 2011 Elsevier Inc. All rights reserved.

Hemocyte-mediated phagocytosis differs between honey bee (Apis mellifera worker castes.

Directory of Open Access Journals (Sweden)

Eva Marit Hystad

Full Text Available Honey bees as other insects rely on the innate immune system for protection against diseases. The innate immune system includes the circulating hemocytes (immune cells that clear pathogens from hemolymph (blood by phagocytosis, nodulation or encapsulation. Honey bee hemocyte numbers have been linked to hemolymph levels of vitellogenin. Vitellogenin is a multifunctional protein with immune-supportive functions identified in a range of species, including the honey bee. Hemocyte numbers can increase via mitosis, and this recruitment process can be important for immune system function and maintenance. Here, we tested if hemocyte mediated phagocytosis differs among the physiologically different honey bee worker castes (nurses, foragers and winter bees, and study possible interactions with vitellogenin and hemocyte recruitment. To this end, we adapted phagocytosis assays, which-together with confocal microscopy and flow cytometry-allow qualitative and quantitative assessment of hemocyte performance. We found that nurses are more efficient in phagocytic uptake than both foragers and winter bees. We detected vitellogenin within the hemocytes, and found that winter bees have the highest numbers of vitellogenin-positive hemocytes. Connections between phagocytosis, hemocyte-vitellogenin and mitosis were worker caste dependent. Our results demonstrate that the phagocytic performance of immune cells differs significantly between honey bee worker castes, and support increased immune competence in nurses as compared to forager bees. Our data, moreover, provides support for roles of vitellogenin in hemocyte activity.

Genotoxic and Cytotoxic Effects on the Immune Cells of the Freshwater Bivalve Dreissena polymorpha Exposed to the Environmental Neurotoxin BMAA.

Science.gov (United States)

Lepoutre, Alexandra; Milliote, Nadia; Bonnard, Marc; Palos-Ladeiro, Mélissa; Rioult, Damien; Bonnard, Isabelle; Bastien, Fanny; Faassen, Elisabeth; Geffard, Alain; Lance, Emilie

2018-03-01

The environmental neurotoxin β- N -Methylamino-l-alanine (BMAA) has been pointed out to be involved in human neurodegenerative diseases. This molecule is known to be bioaccumulated by bivalves. However, little data about its toxic effects on freshwater mussels is available, particularly on the hemolymphatic compartment and its hemocyte cells involved in various physiological processes such as immune defenses, digestion and excretion, tissue repair, and shell production. Here we exposed Dreissena polymorpha to dissolved BMAA, at the environmental concentration of 7.5 µg of /mussel/3 days, during 21 days followed by 14 days of depuration in clear water, with the objective of assessing the BMAA presence in the hemolymphatic compartment, as well as the impact of the hemocyte cells in terms of potential cytotoxicity, immunotoxicity, and genotoxiciy. Data showed that hemocytes were in contact with BMAA. The presence of BMAA in hemolymph did not induce significant effect on hemocytes phagocytosis activity. However, significant DNA damage on hemocytes occurred during the first week (days 3 and 8) of BMAA exposure, followed by an increase of hemocyte mortality after 2 weeks of exposure. Those effects might be an indirect consequence of the BMAA-induced oxidative stress in cells. However, DNA strand breaks and mortality did not persist during the entire exposure, despite the BMAA persistence in the hemolymph, suggesting potential induction of some DNA-repair mechanisms.

Cationic composition and acid-base state of the extracellular fluid, and specific buffer value of hemoglobin from the branchiopod crustacean Triops cancriformis.

Science.gov (United States)

Pirow, Ralph; Buchen, Ina; Richter, Marc; Allmer, Carsten; Nunes, Frank; Günsel, Andreas; Heikens, Wiebke; Lamkemeyer, Tobias; von Reumont, Björn M; Hetz, Stefan K

2009-04-01

Recent insights into the allosteric control of oxygen binding in the extracellular hemoglobin (Hb) of the tadpole shrimp Triops cancriformis raised the question about the physico-chemical properties of the protein's native environment. This study determined the cationic composition and acid-base state of the animal's extracellular fluid. The physiological concentrations of potential cationic effectors (calcium, magnesium) were more than one order of magnitude below the level effective to increase Hb oxygen affinity. The extracellular fluid in the pericardial space had a typical bicarbonate concentration of 7.6 mM but a remarkably high CO(2) partial pressure of 1.36 kPa at pH 7.52 and 20 degrees C. The discrepancy between this high CO(2) partial pressure and the comparably low values for water-breathing decapods could not solely be explained by the hemolymph-sampling procedure but may additionally arise from differences in cardiovascular complexity and efficiency. T. cancriformis hemolymph had a non-bicarbonate buffer value of 2.1 meq L(-1) pH(-1). Hb covered 40-60% of the non-bicarbonate buffering power. The specific buffer value of Hb of 1.1 meq (mmol heme)(-1) pH(-1) suggested a minimum requirement of two titratable histidines per heme-binding domain, which is supported by available information from N-terminal sequencing and expressed sequence tags.

Genotoxic and Cytotoxic Effects on the Immune Cells of the Freshwater Bivalve Dreissena polymorpha Exposed to the Environmental Neurotoxin BMAA

Directory of Open Access Journals (Sweden)

Alexandra Lepoutre

2018-03-01

Full Text Available The environmental neurotoxin β-N-Methylamino-l-alanine (BMAA has been pointed out to be involved in human neurodegenerative diseases. This molecule is known to be bioaccumulated by bivalves. However, little data about its toxic effects on freshwater mussels is available, particularly on the hemolymphatic compartment and its hemocyte cells involved in various physiological processes such as immune defenses, digestion and excretion, tissue repair, and shell production. Here we exposed Dreissena polymorpha to dissolved BMAA, at the environmental concentration of 7.5 µg of /mussel/3 days, during 21 days followed by 14 days of depuration in clear water, with the objective of assessing the BMAA presence in the hemolymphatic compartment, as well as the impact of the hemocyte cells in terms of potential cytotoxicity, immunotoxicity, and genotoxiciy. Data showed that hemocytes were in contact with BMAA. The presence of BMAA in hemolymph did not induce significant effect on hemocytes phagocytosis activity. However, significant DNA damage on hemocytes occurred during the first week (days 3 and 8 of BMAA exposure, followed by an increase of hemocyte mortality after 2 weeks of exposure. Those effects might be an indirect consequence of the BMAA-induced oxidative stress in cells. However, DNA strand breaks and mortality did not persist during the entire exposure, despite the BMAA persistence in the hemolymph, suggesting potential induction of some DNA-repair mechanisms.

Effects of Oral Exposure to Fungicides on Honey Bee Nutrition and Virus Levels.

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Degrandi-Hoffman, Gloria; Chen, Yanping; Watkins Dejong, Emily; Chambers, Mona L; Hidalgo, Geoffrey

2015-12-01

Sublethal exposure to fungicides can affect honey bees (Apis mellifera L.) in ways that resemble malnutrition. These include reduced brood rearing, queen loss, and increased pathogen levels. We examined the effects of oral exposure to the fungicides boscalid and pyraclostrobin on factors affecting colony nutrition and immune function including pollen consumption, protein digestion, hemolymph protein titers, and changes in virus levels. Because the fungicides are respiratory inhibitors, we also measured ATP concentrations in flight muscle. The effects were evaluated in 3- and 7-d-old worker bees at high fungicide concentrations in cage studies, and at field-relevant concentrations in colony studies. Though fungicide levels differed greatly between the cage and colony studies, similar effects were observed. Hemolymph protein concentrations were comparable between bees feeding on pollen with and without added fungicides. However, in both cage and colony studies, bees consumed less pollen containing fungicides and digested less of the protein. Bees fed fungicide-treated pollen also had lower ATP concentrations and higher virus titers. The combination of effects we detected could produce symptoms that are similar to those from poor nutrition and weaken colonies making them more vulnerable to loss from additional stressors such as parasites and pathogens. Published by Oxford University Press on behalf of Entomological Society of America 2015. This work is written by US Government employees and is in the public domain in the US.

Citotoxicidad de extractos de plantas medicinales sobre la línea celular de carcinoma de pulmón humano A549 Cytotoxicity of medicinal plant extracts on the human lung carcinoma cell line A549

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Alexis Díaz García

2011-03-01

Full Text Available OBJETIVO: evaluar el efecto de 10 extractos de plantas medicinales sobre el crecimiento de la línea celular humana de carcinoma de pulmón A549. METODOS: el efecto de los extractos sobre la células tumorales se midió a través de un ensayo colorimétrico mediante el empleo del bromuro de 3-(4,5-dimetil-tiazol-2-yl-2,5-difenil tetrazolio a concentraciones entre 3,9-250 µg/mL durante 72 h y se calculó la concentración citotóxica media para cada uno. RESULTADOS: del total de los extractos evaluados solo cuatro (Parthenium hysterophorus, Bixa orellana, Momordica charantia y Cucurbita maxima evidenciaron concentraciones citotóxicas medias inferiores a 100 µg/mL. Excepto Parthenium hysterophorus, las restantes se emplean en la medicina tradicional para el tratamiento del cáncer. Los extractos de Cecropia peltata, Melia azedarach, Annona glabra, Artemisia absintium, Lepidium virginicum y Bidens pilosa no mostraron efectos citotóxicos significativos. CONCLUSIONES: Los extractos de plantas que se emplean en la medicina tradicional para el tratamiento del cáncer, mostraron citotoxicidad sobre las células tumorales. El conocimiento etnobotánico representa una herramienta importante en la selección de plantas medicinales, en la búsqueda de nuevos compuestos para el tratamiento del cáncer.OBJECTIVES: to evaluate the effect of 10 Cuban medicinal plant extracts on the human lung tumor cell line A549. METHODS: the effect of the plant extracts on tumor cells was determined by a colorimetric assay using the 3-(4,5-Dimethylthiazol-2-yl-2,5-diphenyltetrazolium bromide (MTT at concentrations ranging from 3,9-250 µg/mL for 72 hours and the mean cytotoxic concentration was calculated for each of them. RESULTS: the ethanolic extracts of Parthenium hysterophorus, Bixa orellana, Momordica charantia and Cucurbita maxima showed mean cytotoxic concentrations under 100 µg/mL. Except for P. hysterophorus, the others are used in traditional medicine to fight

Cytogenetic and morphophysiological differences in snail Lymnaea stagnalis (Gastropoda, Pulmonata) from reservoirs of the Chernobyl NPP resettling zone

International Nuclear Information System (INIS)

Koneva, A.Yu.; Afonin, V.Yu.; Dramashka, S.Ya.; Golubew, A.P.

2006-01-01

A cytogenetic analysis of the molluscs obtained from animals of two populations of the Chernobyl NPP zone was carried out. Preservation of the raised number of cells with micronuclei in hemolymph of the molluscs obtained from animals lived under conditions with the highest level of radiation contamination was noted. Interpopulational distinctions in a number of biological parameters were revealed. Away of fertilization which is used for animal production under laboratory conditions affects sensitivity of molluscs to additional acute irradiation was established. (authors)

Fibrinogen-related proteins in ixodid ticks

Czech Academy of Sciences Publication Activity Database

Štěrba, J.; Dupejová, J.; Fišer, M.; Vancová, Marie; Grubhoffer, Libor

2011-01-01

Roč. 4, - (2011), e127 ISSN 1756-3305 R&D Projects: GA MŠk(CZ) LC06009; GA AV ČR KJB600960906; GA ČR GA206/09/1782 Institutional research plan: CEZ:AV0Z60220518 Keywords : AMERICAN DOG TICK * INNATE IMMUNITY * DORIN-M * RHIPICEPHALUS-APPENDICULATUS * MOLECULAR CHARACTERIZATION * DERMACENTOR-VARIABILIS * ORNITHODOROS-MOUBATA * LECTINS * HEMOLYMPH * BINDING Subject RIV: EE - Microbiology, Virology Impact factor: 2.937, year: 2011 http://www.parasitesandvectors.com/content/pdf/1756-3305-4-127.pdf

L-3,4-Dihydroxyphenylalanine (l-DOPA) induces neuroendocrinological, physiological, and immunological regulation in white shrimp, Litopenaeus vannamei.

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Mapanao, Ratchaneegorn; Kuo, Hsin-Wei; Chang, Chin-Chuan; Liu, Kuan-Fu; Cheng, Winton

2018-03-01

L-3,4-Dihydroxyphenylalanine (l-DOPA) is a precursor for dopamine (DA) synthesis. Assessments were conducted to analyze the effects of l-DOPA on mediating regulation of neuroendocrinological, immunological, and physiological parameters in the shrimp, Litopenaeus vannamei when they were individually injected with 0.01 N HCl or l-DOPA at 0.5 or 1.0 μmol shrimp -1 for 60, 120, and 240 min. For catecholamine synthesis evaluation, tyrosine hydroxylase (TH) and DA beta hydroxylase (DBH) activities, l-DOPA, DA, and norepinephrine (NE) levels in hemolymph were determined. The total hemocyte count (THC), differential hemocyte count (DHC), phenoloxidase (PO) activity, respiratory bursts (RBs), superoxide dismutase (SOD) activity, phagocytic activity, and clearance efficiency in response to the pathogen, Vibrio alginolyticus were assessed for immune responses, and plasma glucose and lactate levels were for physiological response. Results showed that the TH activity, THC, hyaline cells (HCs), and semigranular cells (SGCs) at 120 min, DA levels at 60-240 min, PO activity in hemocytes per 50 μL of hemolymph at 60-120 min, and PO activity per granulocyte (granular cells (GCs) + SGCs) at 60 min significantly increased, but TH activity, l-DOPA levels, GCs, SGCs, and respiratory bursts in hemocytes per 10 μL of hemolymph at 60 min, respiratory bursts per hemocyte and SOD activity at 120 min, phagocytic activity at 60-240 min, and the clearance efficiency at 60-120 min significantly decreased in shrimp injected with l-DOPA at 1.0 μmol shrimp -1 . In another experiment, 60 min after shrimp had received l-DOPA at 0.5 or 1.0 μmol shrimp -1 , they were challenged with an injection of V. alginolyticus at 2 × 10 5  colony-forming units (cfu) shrimp -1 . The injection of l-DOPA at 1.0 μmol shrimp -1 also significantly increased the cumulative mortality of shrimp by 16.7%, compared to the HCl-challenged control after 120 h. These results suggest

Vertebrate estrogen regulates the development of female characteristics in silkworm, Bombyx mori.

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Shen, Guanwang; Lin, Ying; Yang, Congwen; Xing, Runmiao; Zhang, Haiyan; Chen, Enxiang; Han, Chaoshan; Liu, Hongling; Zhang, Weiwei; Xia, Qingyou

2015-01-01

The vertebrate estrogens include 17-β-estradiol (E2), which has an analog in silkworm ovaries. In this study, the Bombyx mori vitellogenin gene (BmVg) was used as a biomarker to analyze the function of the E2 in silkworm. In most oviparous animals, Vg has female-specific expression. However, BmVg expression was also detected in B. mori males. Stage specific fluctuation of BmVg expression was similar in males and females, but expression levels in males were lower than in females. E2 treatment by injection or feeding of male larvae in the final instar stage induced and stimulated male BmVg transcription and protein synthesis. When silkworm ovary primordia were transplanted into males, BmVg was induced in male fat bodies. Transplanted ovaries primordia were also able to develop into ovaries and produce mature eggs. When females were treated with E2 promoted BmVg/BmVn protein accumulation in hemolymph, ovaries and eggs. However, BmVg transcription was decreased in female fat bodies. An E2 analog was identified in the hemolymph of day 3 wandering silkworms using high-performance liquid chromatography. Estradiol titers from fifth late-instar larvae to pupal stage were determined by enzyme-linked immunosorbent assay. The results suggested that silkworms synthesized a vertebrate E2 analog. This study found that E2 promoted the synthesis of BmVg, a female typical protein in silkworms. Copyright © 2014 Elsevier Inc. All rights reserved.

The Silencing of a 14-3-3ɛ Homolog in Tenebrio molitor Leads to Increased Antimicrobial Activity in Hemocyte and Reduces Larval Survivability.

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Seo, Gi Won; Jo, Yong Hun; Seong, Jeong Hwan; Park, Ki Beom; Patnaik, Bharat Bhusan; Tindwa, Hamisi; Kim, Sun-Am; Lee, Yong Seok; Kim, Yu Jung; Han, Yeon Soo

2016-08-20

The 14-3-3 family of phosphorylated serine-binding proteins acts as signaling molecules in biological processes such as metabolism, division, differentiation, autophagy, and apoptosis. Herein, we report the requirement of 14-3-3ɛ isoform from Tenebrio molitor (Tm14-3-3ɛ) in the hemocyte antimicrobial activity. The Tm14-3-3ɛ transcript is 771 nucleotides in length and encodes a polypeptide of 256 amino acid residues. The protein has the typical 14-3-3 domain, the nuclear export signal (NES) sequence, and the peptide binding residues. The Tm14-3-3ɛ transcript shows a significant three-fold expression in the hemocyte of T. molitor larvae when infected with Escherichia coli Tm14-3-3ɛ silenced larvae show significantly lower survival rates when infected with E. coli. Under Tm14-3-3ɛ silenced condition, a strong antimicrobial activity is elicited in the hemocyte of the host inoculated with E. coli. This suggests impaired secretion of antimicrobial peptides (AMP) into the hemolymph. Furthermore, a reduction in AMP secretion under Tm14-3-3ɛ silenced condition would be responsible for loss in the capacity to kill bacteria and might explain the reduced survivability of the larvae upon E. coli challenge. This shows that Tm14-3-3ɛ is required to maintain innate immunity in T. molitor by enabling antimicrobial secretion into the hemolymph and explains the functional specialization of the isoform.

The Silencing of a 14-3-3ɛ Homolog in Tenebrio molitor Leads to Increased Antimicrobial Activity in Hemocyte and Reduces Larval Survivability

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Gi Won Seo

2016-08-01

Full Text Available The 14-3-3 family of phosphorylated serine-binding proteins acts as signaling molecules in biological processes such as metabolism, division, differentiation, autophagy, and apoptosis. Herein, we report the requirement of 14-3-3ɛ isoform from Tenebrio molitor (Tm14-3-3ɛ in the hemocyte antimicrobial activity. The Tm14-3-3ɛ transcript is 771 nucleotides in length and encodes a polypeptide of 256 amino acid residues. The protein has the typical 14-3-3 domain, the nuclear export signal (NES sequence, and the peptide binding residues. The Tm14-3-3ɛ transcript shows a significant three-fold expression in the hemocyte of T. molitor larvae when infected with Escherichia coli Tm14-3-3ɛ silenced larvae show significantly lower survival rates when infected with E. coli. Under Tm14-3-3ɛ silenced condition, a strong antimicrobial activity is elicited in the hemocyte of the host inoculated with E. coli. This suggests impaired secretion of antimicrobial peptides (AMP into the hemolymph. Furthermore, a reduction in AMP secretion under Tm14-3-3ɛ silenced condition would be responsible for loss in the capacity to kill bacteria and might explain the reduced survivability of the larvae upon E. coli challenge. This shows that Tm14-3-3ɛ is required to maintain innate immunity in T. molitor by enabling antimicrobial secretion into the hemolymph and explains the functional specialization of the isoform.

The Gut Entomotype of Red Palm Weevil Rhynchophorus ferrugineus Olivier (Coleoptera: Dryophthoridae and Their Effect on Host Nutrition Metabolism

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Abrar Muhammad

2017-11-01

Full Text Available For invasive insects, the potential roles of gut microbiota in exploiting new food resources and spreading remain elusive. Red palm weevil (RPW, Rhynchophorus ferrugineus Olivier, is an invasive destructive pest which feeds on nutrient-poor tender tissues and has caused extensive mortality of palm trees. The microbes associated with insects can improve their nutrition assimilation. However, experimental evidence on the interactions between RPW and its gut microbiota is still absent. The aim of this study is to determine the dynamics changes and the bacterial entomotype in the RPW gut and its potential physiological roles. Here, we confirmed RPW harbors a complex gut microbiota mainly constituted by bacteria in the families Enterobacteriaceae, Lactobacillaceae, Entomoplasmataceae, and Streptococcaceae. RPW gut microbiota exhibited a highly stable microbial community with low variance in abundance across different life stages and host plants. Furthermore, the abundance of Enterobacteriaceae was markedly increased but that of Acetobacteraceae was reduced significantly after administration of antibiotics. Although no significant effects were found on the body weight gain of RPW larvae, these alterations dramatically decreased the concentration of hemolymph protein and glucose while that of hemolymph triglyceride increased. In the gut of wild-caught RPW larvae, seven bacterial species in the genera Klebsiella, Serratia, Enterobacter, and Citrobacter were shown to have an ability to degrade cellulose. Together, RPW accommodate a stable gut microbiota which can degrade plant polysaccharides and confer their host optimal adaptation to its environment by modulating its metabolism.

Complement C3 gene: Expression characterization and innate immune response in razor clam Sinonovacula constricta.

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Peng, Maoxiao; Niu, Donghong; Wang, Fei; Chen, Zhiyi; Li, Jiale

2016-08-01

Complement component 3 (C3) is central to the complement system, playing an important role in immune defense, immune regulation and immune pathology. Several C3 genes have been characterized in invertebrates but very few in shellfish. The C3 gene was identified from the razor clam Sinonovacula constricta, referred to here as Sc-C3. It was found to be highly homologous with the C3 gene of Ruditapes decussatus. All eight model motifs of the C3 gene were found to be included in the thiolester bond and the C345C region. Sc-C3 was widely expressed in all healthy tissues with expression being highest in hemolymph. A significant difference in expression was revealed at the umbo larvae development stage. The expression of Sc-C3 was highly regulated in the hemolymph and liver, with a distinct response pattern being noted after a challenge with Micrococcus lysodeikticus and Vibrio parahemolyticus. It is therefore suggested that a complicated and unique response pathway may be present in S. constricta. Further, serum of S. constricta containing Sc-C3 was extracted. This was activated by LPS or bacterium for verification for function. The more obvious immune function of Sc-C3 was described as an effective membrane rupture in hemocyte cells of rabbit, V. parahemolyticus and Vibrio anguillarum. Thus, Sc-C3 plays an essential role in the immune defense of S. constricta. Copyright © 2016 Elsevier Ltd. All rights reserved.

Short-term heat shock affects the course of immune response in Galleria mellonella naturally infected with the entomopathogenic fungus Beauveria bassiana.

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Vertyporokh, Lidiia; Taszłow, Paulina; Samorek-Pieróg, Małgorzata; Wojda, Iwona

2015-09-01

We aimed to investigate how exposition of infected insects to short-term heat shock affects the biochemical and molecular aspects of their immune response. Galleria mellonella larvae were exposed to 43°C for 15min, at the seventy second hour after natural infection with entomopathogenic fungus Beauveria bassiana. As a result, both qualitative and quantitative changes in hemolymph protein profiles, and among them infection-induced changes in the amount of apolipophorin III (apoLp-III), were observed. Heat shock differently affects the expression of the tested immune-related genes. It transiently inhibits expression of antifungal peptides gallerimycin and galiomicin in both the fat body and hemocytes of infected larvae. The same, although to a lesser extent, concerned apoLp-III gene expression and was observed directly after heat shock. Nevertheless, in larvae that had recovered from heat shock, apoLp-III expression was higher in comparison to unshocked larvae in the fat body but not in hemocytes, which was consistent with the higher amount of this protein detected in the hemolymph of the infected, shocked larvae. Furthermore, lysozyme-type activity was higher directly after heat shock, while antifungal activity was significantly higher also in larvae that had recovered from heat shock, in comparison to the respective values in their non-shocked, infected counterparts. These results show how changes in the external temperature modulate the immune response of G. mellonella suffering from infection with its natural pathogen B. bassiana. Copyright © 2015 Elsevier Inc. All rights reserved.

Molecular cloning and responsive expression to injury stimulus of a defender against cell death 1 (DAD1) gene from bay scallops Argopecten irradians.

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Zhu, Ling; Song, Linsheng; Zhang, Huan; Zhao, Jianmin; Li, Chenghua; Xu, Wei

2008-06-01

Apoptosis is an active process of cell death, which is an integral part of growth and development in multicellular organisms. The defender against cell death 1 (DAD1), the regulatory protein to inhibit the apoptosis process, was first cloned from the bay scallop Argopecten irradians by randomly sequencing a whole tissue cDNA library and rapid amplification of cDNA end (RACE). The full-length cDNA of the A. irradians DAD1 was 607 bp, consist of a 5'-terminal untranslated region (UTR) of 63 bp, a 3'-terminal UTR of 205 bp with a canonical polyadenylation signal sequence AATAAA and a poly (A) tail, and an open reading frame of 339 bp. The deduced amino acid sequence of the A. irradians DAD1 showed 75.5% identity to Araneus ventricosus, 74.5% to Drosophila melanogaster, and 73.6% to Homo sapiens, Sus scrofa, Mesocricetus auratus, Rattus norvegicus and Mus musculus. Excluding the Saccharomyces cerevisiae DAD1 homologue, all animal DAD1 including A. irradians DAD1 homologue formed a subgroup and all plant DAD1 proteins formed another subgroup in the phylogenetic analysis. The A. irradians DAD1 was expressed in all examined tissues including adductor muscle, mantle, gills, digestive gland, gonad and hemolymph, suggesting that A. irradians DAD1 is expressed in most body tissues. Furthermore, the mRNA expression levels of A. irradians DAD1 gene of hemolymph were particularly high after injury, suggesting that the gene is responsive to injury stimuli.

Methyl Farnesoate Plays a Dual Role in Regulating Drosophila Metamorphosis

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Wen, Di; Rivera-Perez, Crisalejandra; Abdou, Mohamed; Jia, Qiangqiang; He, Qianyu; Liu, Xi; Zyaan, Ola; Xu, Jingjing; Bendena, William G.; Tobe, Stephen S.; Noriega, Fernando G.; Palli, Subba R.; Wang, Jian; Li, Sheng

2015-01-01

Corpus allatum (CA) ablation results in juvenile hormone (JH) deficiency and pupal lethality in Drosophila. The fly CA produces and releases three sesquiterpenoid hormones: JH III bisepoxide (JHB3), JH III, and methyl farnesoate (MF). In the whole body extracts, MF is the most abundant sesquiterpenoid, followed by JHB3 and JH III. Knockout of JH acid methyl transferase (jhamt) did not result in lethality; it decreased biosynthesis of JHB3, but MF biosynthesis was not affected. RNAi-mediated reduction of 3-hydroxy-3-methylglutaryl CoA reductase (hmgcr) expression in the CA decreased biosynthesis and titers of the three sesquiterpenoids, resulting in partial lethality. Reducing hmgcr expression in the CA of the jhamt mutant further decreased MF titer to a very low level, and caused complete lethality. JH III, JHB3, and MF function through Met and Gce, the two JH receptors, and induce expression of Kr-h1, a JH primary-response gene. As well, a portion of MF is converted to JHB3 in the hemolymph or peripheral tissues. Topical application of JHB3, JH III, or MF precluded lethality in JH-deficient animals, but not in the Met gce double mutant. Taken together, these experiments show that MF is produced by the larval CA and released into the hemolymph, from where it exerts its anti-metamorphic effects indirectly after conversion to JHB3, as well as acting as a hormone itself through the two JH receptors, Met and Gce. PMID:25774983

Osmotic concentration in three races of honey bee, Apis mellifera L. under environmental conditions of arid zone.

Science.gov (United States)

Ali, Hussain; Alqarni, Abdulaziz S; Owayss, Ayman A; Hassan, Awad M; Smith, Brian H

2017-07-01

Hemolymph osmolarity has great effect on honey bee health, especially in arid and semi-arid zones. It regulates water and nutrients in stressed tissues. Osmotic concentration in three races ( Apis mellifera ligustica , A. m. carnica and A. m. jemenitica ) of Apis mellifera was tested in central Saudi Arabia during spring and summer seasons in 2015. Newly emerged bee workers were first marked and later their hemolymph was extracted after intervals of 1, 5, 10, 15, 20 and 25 days. A significant positive correlation between age and osmolarity was found in all three races during spring and summer seasons. The lowest combined osmotic concentration for all three races was found after 1 day interval, while the highest osmotic concentration was recorded after 25 days. Among all races, A. m. ligustica showed significantly high osmotic concentration after 25 days in spring and summer seasons as compared to the other two races. Only A. m. jemenitica showed similar osmotic concentration after 10 and 15 days in both spring and summer seasons compared to other two races. Mean osmotic concentration of all three races was significantly different after 20 and 25 days in spring and summer seasons. Overall mean recorded during summer was significantly higher than the mean of spring season. Combined osmotic concentration in young drones of all races was significantly lower than that of old drones during spring and summer seasons.

Unveiling the oxidative metabolism of Rhipicephalus microplus (Acari: Ixodidae) experimentally exposed to entomopathogenic fungi.

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Tunholi-Alves, Vinícius Menezes; Tunholi Alves, Victor Menezes; da Silva, Jairo Pinheiro; Nora Castro, Rosane; Salgueiro, Fernanda Barbosa; Perinotto, Wendell Marcelo de Souza; Gôlo, Patrícia Silva; Camargo, Mariana Guedes; Angelo, Isabele da Costa; Bittencourt, Vânia Rita Elias Pinheiro

2016-10-01

Rhipicephalus microplus is an important tick in tropical regions due to the high economic losses caused by its parasitism. Metarhizium anisopliae and Beauveria bassiana are well-known entomopathogenic fungi that can afflict R. microplus ticks. The development of new targets and strategies to control this parasite can be driven by studies of this tick's physiology. Recently, it was reported that when exposed to adverse physiological conditions, ticks can activate fermentative pathways, indicating transition from aerobic to anaerobic metabolism. Nevertheless, the precise mechanism by which entomopathogenic fungi influence R. microplus metabolism has not been clarified, limiting understanding of the tick-fungus association. Thus, the present study aimed to evaluate the effect of infection of ticks by M. anisopliae and B. bassiana on the amount of selected carboxylic acids present in the hemolymph, enabling increased understanding of changes previously reported. The results showed preservation in the concentrations of oxalic, lactic, and pyruvic acids in the hemolymph 24 and 48 h after dropping from cattle; while there were variations in the concentration of these carboxylic acids after infection of female ticks to M. anisopliae and B. bassiana. Significant increases were observed in the concentration of oxalic and lactic acids and significant reduction of pyruvic acid for both observation times (24 and 48 h) after infection by entomopathogenic fungi. These results indicate that B. bassiana and M. anisopliae infection alters the basal metabolism of R. microplus females, resulting in the activation of fermentative pathways.

Entomopathogenic Fungi: New Insights into Host-Pathogen Interactions.

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Butt, T M; Coates, C J; Dubovskiy, I M; Ratcliffe, N A

2016-01-01

Although many insects successfully live in dangerous environments exposed to diverse communities of microbes, they are often exploited and killed by specialist pathogens. Studies of host-pathogen interactions (HPI) provide valuable insights into the dynamics of the highly aggressive coevolutionary arms race between entomopathogenic fungi (EPF) and their arthropod hosts. The host defenses are designed to exclude the pathogen or mitigate the damage inflicted while the pathogen responds with immune evasion and utilization of host resources. EPF neutralize their immediate surroundings on the insect integument and benefit from the physiochemical properties of the cuticle and its compounds that exclude competing microbes. EPF also exhibit adaptations aimed at minimizing trauma that can be deleterious to both host and pathogen (eg, melanization of hemolymph), form narrow penetration pegs that alleviate host dehydration and produce blastospores that lack immunogenic sugars/enzymes but facilitate rapid assimilation of hemolymph nutrients. In response, insects deploy an extensive armory of hemocytes and macromolecules, such as lectins and phenoloxidase, that repel, immobilize, and kill EPF. New evidence suggests that immune bioactives work synergistically (eg, lysozyme with antimicrobial peptides) to combat infections. Some proteins, including transferrin and apolipophorin III, also demonstrate multifunctional properties, participating in metabolism, homeostasis, and pathogen recognition. This review discusses the molecular intricacies of these HPI, highlighting the interplay between immunity, stress management, and metabolism. Increased knowledge in this area could enhance the efficacy of EPF, ensuring their future in integrated pest management programs. Copyright © 2016 Elsevier Inc. All rights reserved.

Late glacial and Holocene history of the dry forest area in south Colombian Cauca Valley from sites Quilichao and la Teta

International Nuclear Information System (INIS)

Berrio, Juan Carlos; Hooghiemstra, Henry; Marchant, Rob; Rangel Orlando

2002-01-01

Two sedimentary cores with records of pollen and charcoal content within a chronology provided by radiocarbon are presented from the southern Cauca Valley in Colombia (1020 m). These records document the late glacial and Holocene dry forest vegetation, fire and environment history. Specifically, core Quilichao -1 (640 cm; 3 degrade 6' N, 76 degrade 31' W) represents the periods of 13/150-7720 14 C yr BP and following a hiatus from 2880 14 C yr BP to recent. Core la Teta - 2 (250 cm; 3 degrade 5' N, 76 degrade 32' W) provides a record from 8700 14 C yr BP around 13/150 21 4 C yr BP Quilichao shown an active late glacial drainage system and presence of dry forest. From 11/465-10/520 14 C yr BP dry forest consists mainly of Crotalaria Moraceae Urticaceae, Melastomataceae, Piper and low stature trees, such as Acalypha, Alchornea, Cecropia and Celtis. At higher elevation on the slopes Andean forest with Quercus, Hedyosmum, Myrica and Alnus are common

Palynological Origin, Phenolic Content, and Antioxidant Properties of Honeybee-Collected Pollen from Bahia, Brazil

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Francisco A. R. Santos

2012-02-01

Full Text Available The aim of this study was to determine the palynological origin, phenolic and flavonoid content, and antioxidant properties of twenty-five samples of bee pollen harvested during a nine-month period (February–November from the Canavieiras municipality (northeastern Brazil. Of the 25 samples analyzed, only two (February 01 and 02 were heterofloral. The predominant pollens in the samples analyzed during that month were: Cecropia, Eucalyptus, Elaeis, Mimosa pudica, Eupatorium, and Scoparia. Ethyl acetate fractions were analyzed by HPLC-DAD. The flavonoids isoquercetin, myricetin, tricetin, quercetin, luteolin, selagin, kaempferol, and isorhamnetin were detected. The flavonoid present in all 22 samples was isolated and identified as isorhamnetin 3-O-b-neohesperidoside. The total phenolic contents determined using the Folin-Ciocalteu reagent ranged from 41.5 to 213.2 mg GAE/g. Antioxidant activities based on the 1,1-diphenyl-2-picryl hydrazyl (DPPH, 2,2-azinobis 3-ethylbenzothiozoline-6-sulfonic acid (ABTS, and Fe2+ ion chelating activity assays were observed for all extracts, and correlated with the total phenolic content.

ANÁLISIS DE LAS FUERZAS DEL MERCADO DENDROENERGÉTICO EN LA VEREDA CAJETE DEL MUNICIPIO DE POPAYÁN

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ALEX ENRIQUE ORDOÑEZ HOYOS

2017-07-01

Full Text Available This paper presents an analysis of the wood energy market forces in the village of Cajete the municipality of Popayan. The methodology led to the realization of a random sampling of wood used as woodfuel and proceeded on which the application of a stratified survey of 64 selected users according to previously identified patterns of consumption. They were found to be the most commonly used species such as fuelwood in the sidewalk coffee (Coffea arabica with 22%, pine (Pinus sp 18%, Cúcharo (Myrsine guianensis 15% and the Yarumo (Cecropia peltata 15%. The total demand for fuelwood estimated for the sidewalk was 122,7 m3 per month, being households that represent the highest proportion 96,2%, while ovens and restaurants represent 3,8%. With respect to the offer found that 63% of users collect firewood from the forests and farms of the area, while 37% buy the product that comes mainly from sawmills located in the urban area of the municipality of Popayán.

SH2B Regulation of Growth, Metabolism, and Longevity in Both Insects and Mammals

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Song, Wei; Ren, Decheng; Li, Wenjun; Jiang, Lin; Cho, Kae Won; Huang, Ping; Fan, Chen; Song, Yiyun; Liu, Yong; Rui, Liangyou

2010-01-01

SH2B1 is a key regulator of body weight in mammals. Here we identified dSH2B as the Drosophila homolog of SH2B1. dSH2B bound to Chico and directly promoted insulin-like signaling. Disruption of dSH2B decreased insulin-like signaling and somatic growth in flies. dSH2B deficiency also increased hemolymph carbohydrate levels, whole body lipid levels, lifespan, and resistance to starvation and oxidative stress. Systemic overexpression of dSH2B resulted in opposite phenotypes. dSH2B overexpression...

Delayed Presentation of a Chronic Morel-Lavallée Lesion.

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Christian, David; Leland, Hyuma A; Osias, Walter; Eberlin, Seth; Howell, Lori

2016-07-01

Morel-Lavellée lesions are soft tissue degloving injuries resulting from shearing trauma that induces separation of the superficial and deep fascias creating a potential space that becomes filled with hemolymph. Here we present a case of a 28-year-old male presenting with a persistent Type I Morel-Lavallée lesion 2.5 years after an automobile versus pedestrian accident. These lesions can be visualized via computed tomography, plain film and ultrasound, but magnetic resonance imaging is the modality of choice for their identification and characterization.

Análise fitossociológica de um remanescente de vegetação na microbacia do Córrego Criminoso (Bacia do Rio Taquari, Coxim, MS, Brasil: subsídios para a recomposição da vegetação Phytosociological analysis of a vegetation remnant in the Córrego Criminoso Basin (Taquari River Basin, Coxim District, Mato Grosso do Sul, Brazil: data for vegetation recovery studies

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Adriana Maria Güntzel

2011-09-01

Full Text Available O objetivo deste estudo foi realizar a análise florística e fitossociológica preliminar em um remanescente de vegetação na área de entorno do Córrego Criminoso, visando subsidiar futuros projetos de recomposição da vegetação. A área total do remanescente foi determinada com o auxílio de GPS e dos programas Pathfinder e Autocad. Amostragens de indivíduos com CAP maior ou igual a 20 cm foram realizadas em 20 parcelas de 300 m² distribuídas na área. Foram encontradas 26 famílias botânicas contendo 36 gêneros e 49 espécies. A família Leguminosae foi a mais rica em espécies, seguida da família Annonaceae. A espécie Xylopia aromatica foi a mais freqüente, apresentando valores de IVI e IVC de 99,77 e 85,73, respectivamente. Observou-se que mais de 50% das espécies presentes na área são pioneiras ou secundárias, sendo cinco consideradas típicas de matas ciliares: Tapirira guianenses, Cecropia pachystachya, Terminalia argentea, Ocotea pulchellae e Luehea grandiflora. O remanescente de vegetação do entorno do Córrego Criminoso encontra-se fortemente degradado, necessitando de ações de manejo específicos nos fragmentos vegetados e nas áreas destituídas de vegetação, cujas recomendações são propostas.The aim of this study was to complete a preliminary, floristic, phytosociological analyses of a vegetation remnant in the Criminoso Stream Basin, in order to provide data for future projects focusing on vegetation recovery. The total area of the remnant was determined using a GPS and the software Pathfinder and Autocad. Samples of individuals larger or equal to 20 cm CBH were made in 20 quadrats that were 300 m². Thirty-six genera and 49 species, within 26 botanical families, were found. The family Leguminosae was the richest in species, followed by the Annonaceae. Xylopia aromatica was the most frequent species, presenting values of IVI and IVC of 99.77 and 85.73, respectively. More than 50% of the species present

Comparison of the diet of Alouatta caraya (Primates: Atelidae between a riparian island and mainland on the Upper Parana River, southern Brazil Comparação da dieta de Alouatta caraya (Primates: Atelidae em mata ciliar insular e continental na região do Alto rio Paraná, Sul do Brasil

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Gabriela Ludwig

2008-09-01

Full Text Available Howler monkeys (Alouatta Lacèpéde, 1799 are folivores-frugivores with flexible diets depending on conditions. Here, we compare the diets of Alouatta caraya (Humboldt, 1812 in two riparian forests (island and mainland, in Porto Rico region, Upper Paraná River, Southern Brazil. Howlers were followed from October 2004 to September 2005 in the riparian forest of a 1,050 ha island and in the continuous riparian forest on the mainland (left bank of the river. The "scan sampling" method with instantaneous samples every 15 minutes was used. Besides vines, diet breadth was similar: 17 species consumed on the island versus 16 species on the mainland. Both consumed leaves followed by fruits more than any other food type (leaves: island - 65%, mainland - 49%, fruits: island - 24%, mainland - 46%. Even though the plant Cecropia pachystachya Trécul is less abundant in the mainland it was the item most consumed in both locations all year long, which suggests its importance for the howlers. Diet also varied both seasonally and between the island and mainland, apparently following changes in local abundance of each item and due to plant phenologies.Os bugios (Alouatta Lacèpéde, 1799 são primatas folívoros-frugívoros que podem adaptar suas dietas dependendo das condições do ambiente. O presente trabalho compara a dieta de grupos de Alouatta caraya (Humboldt, 1812 presentes em matas ciliares estruturalmente diferentes (ilha e continente, na região de Porto Rico, Alto rio Paraná, sul do Brasil. Os bugios-pretos foram seguidos de outubro de 2004 a setembro de 2005 em mata ciliar de uma ilha fluvial de 1050 ha e em mata ciliar contínua da margem esquerda do rio. Foi utilizado o método "scan sampling" em amostragens instantâneas com intervalos de 15 minutos. Além das lianas, 17 espécies foram consumidas na ilha versus 16 espécies no continente. Ambos os grupos consumiram mais folhas seguido de frutos do que qualquer outro tipo de alimento (folhas

Ocorrência do pólen de Podocarpus sp. (Podocarpaceae nas coletas de Frieseomelitta varia Lepeletier 1836 (Apidae: Meliponinae em uma área de Manaus, AM, Brasil Occurrence of Podocarpus sp. (Podocarpaceae pollen on Frieseomelitta varia Lepeletier 1836 (Apidae: Meliponinae in Manaus, Amazonas, Brazil

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Antonio Carlos Marques-Souza

2010-06-01

Full Text Available Durante o período de doze meses, o pólen transportado por Frieseomelitta varia Lepeletier 1836 foi coletado das corbículas das operárias, logo após o fechamento da entrada das colméias. Feita a identificação polínica dos grãos de pólen e sua frequência mensal nas amostras, constatou-se que as espécies das famílias mais visitadas pela ordem de coleta, foram: Cecropiaceae, Sapotaceae, Myrtaceae e Moraceae. As operárias coletaram o pólen de 79 espécies de plantas distribuídas em 60 gêneros e 37 famílias, sendo as mais frequentes: Cecropia sp. visitada o ano todo com atratividade em janeiro de 82,03%, Morus sp. com atratividade de 37,46% em dezembro; Myrcia amazonica DC. com atratividade em abril de 32,34% e Pouteria macrophylla (A.DC. Eyma em junho com atratividade de 36,54%. Quanto a Podocarpus sp. esse é o primeiro relato da presença do pólen dessa espécie em coletas de meliponíneos, o que não deixa de ser um fato curioso, uma vez que se trata de uma gimnosperma encontrada em áreas específicas da região amazônica e que apresentou atratividade para Frieseomelitta varia Lepeletier 1836 de 4,94% no mês de março.Over a 12 month period, pollen was sampled from the corbiculae of workers of the stingless bee Frieseomelitta varia Lepeletier 1836. Collections were obtained immediately after sealing the hives, located in a forest fragment in the city of Manaus. Pollen was identified and monthly frequency of each type was tallied. The most-visited plant families in decreasing order of frequency were Cecropiaceae, Sapotaceae, Myrtaceae and Moraceae. Identified pollen was harvested from 79 plant species in 60 genera and 37 families. The most frequent species were: Cecropia sp., visited year-round but with highest attractiveness (82% in January; Morus sp., with peak attractiveness (37% in December; Myrcia amazonica DC. with 32% peak attractiveness in April; and Pouteria macrophylla (A.DC. Eyma, with 37% peak attractiveness in

Effect of Propolis Oral Intake on Physiological Condition of Young Worker Honey Bees, Apis Mellifera L.

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Damiani Natalia

2017-12-01

Full Text Available Honey bees collect resin from various plant species and transform it into propolis that is incorporated into the nest. The role of resins in the bee health field is poorly understood. The aim was to evaluate the effects of forced consumption of propolis on the physiological condition and short-term survival of Apis mellifera worker bees. It was tested if the number of circulating hemocytes in hemolymph, the abdominal fat bodies and the hypopharyngeal glands development were affected by the feeding with propolis extracts in laboratory conditions during the warm and the cold seasons. Propolis added to sugar candy was consumed by workers for fourteen days without affecting the bee survival. The number of circulating hemocytes in hemolymph remained constant despite the differential diet during the experiment. However, the development of fat bodies and hypopharyngeal glands was altered by propolis ingestion. The abdominal fat body development in winter bees diminished after fourteen days of propolis consumption, while it increased in summer bees. The hypopharyngeal gland development decreased for the assayed period in workers from both seasons. Our results encourage us to continue exploring this research field and learn how long-term forced ingestion of a plant-derived compound, a non-nutritive substance, can modify physiological bee parameters. A broader understanding of the multiple roles of propolis in the health of the honey bee colonies could be obtained by studying the ways in which it is processed and metabolized and the effect that generates in another physiological responses.

Cecropins as a marker of Spodoptera frugiperda immunosuppression during entomopathogenic bacterial challenge.

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Duvic, B; Jouan, V; Essa, N; Girard, P-A; Pagès, S; Abi Khattar, Z; Volkoff, N-A; Givaudan, A; Destoumieux-Garzon, D; Escoubas, J-M

2012-06-01

An antimicrobial peptide (AMP) of the cecropin family was isolated by HPLC from plasma of the insect pest, Spodoptera frugiperda. Its molecular mass is 3910.9 Da as determined by mass spectrometry. Thanks to the EST database Spodobase, we were able to describe 13 cDNAs encoding six different cecropins which belong to the sub-families CecA, CecB, CecC and CecD. The purified peptide identified as CecB1 was chemically synthesized (syCecB1). It was shown to be active against Gram-positive and Gram-negative bacteria as well as fungi. Two closely related entomopathogenic bacteria, Xenorhabdus nematophila F1 and Xenorhabdus mauleonii VC01(T) showed different susceptibility to syCecB1. Indeed, X. nematophila was sensitive to syCecB1 whereas X. mauleonii had a minimal inhibitory concentration (MIC) eight times higher. Interestingly, injection of live X. nematophila into insects did not induce the expression of AMPs in hemolymph. This effect was not observed when this bacterium was heat-killed before injection. On the opposite, both live and heat-killed X. mauleonii induced the expression of AMPs in the hemolymph of S. frugiperda. The same phenomenon was observed for another immune-related protein lacking antimicrobial activity. Altogether, our data suggest that Xenorhabdus strains have developed different strategies to supplant the humoral defense mechanisms of S. frugiperda, either by increasing their resistance to AMPs or by preventing their expression during such host-pathogen interaction. Copyright © 2012 Elsevier Ltd. All rights reserved.

OpnS, an outer membrane porin of Xenorhabdus nematophila, confers a competitive advantage for growth in the insect host.

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van der Hoeven, Ransome; Forst, Steven

2009-09-01

The gammaproteobacterium Xenorhabdus nematophila engages in a mutualistic association with an entomopathogenic nematode and also functions as a pathogen toward different insect hosts. We studied the role of the growth-phase-regulated outer membrane protein OpnS in host interactions. OpnS was shown to be a 16-stranded beta-barrel porin. opnS was expressed during growth in insect hemolymph and expression was elevated as the cell density increased. When wild-type and opnS deletion strains were coinjected into insects, the wild-type strain was predominantly recovered from the insect cadaver. Similarly, an opnS-complemented strain outcompeted the DeltaopnS strain. Coinjection of the wild-type and DeltaopnS strains together with uncolonized nematodes into insects resulted in nematode progeny that were almost exclusively colonized with the wild-type strain. Likewise, nematode progeny recovered after coinjection of a mixture of nematodes carrying either the wild-type or DeltaopnS strain were colonized by the wild-type strain. In addition, the DeltaopnS strain displayed a competitive growth defect when grown together with the wild-type strain in insect hemolymph but not in defined culture medium. The DeltaopnS strain displayed increased sensitivity to antimicrobial compounds, suggesting that deletion of OpnS affected the integrity of the outer membrane. These findings show that the OpnS porin confers a competitive advantage for the growth and/or the survival of X. nematophila in the insect host and provides a new model for studying the biological relevance of differential regulation of porins in a natural host environment.

Differential sensitivity of Chironomus and human hemoglobin to gamma radiation

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Gaikwad, Pallavi S. [Stress Biology Research Laboratory, Department of Zoology, Savitribai Phule University, Pune, 411007 (India); Molecular Biology Division, Bhabha Atomic Research Centre, Trombay, Mumbai, 400085 (India); Panicker, Lata [Solid State Physics Division, Bhabha Atomic Research Centre, Trombay, Mumbai, 400085 (India); Mohole, Madhura; Sawant, Sangeeta [Bioinformatics Center, Savitribai Phule Pune University, Pune, 411007 (India); Mukhopadhyaya, Rita [Molecular Biology Division, Bhabha Atomic Research Centre, Trombay, Mumbai, 400085 (India); Nath, Bimalendu B., E-mail: bbnath@gmail.com [Stress Biology Research Laboratory, Department of Zoology, Savitribai Phule University, Pune, 411007 (India)

2016-08-05

Chironomus ramosus is known to tolerate high doses of gamma radiation exposure. Larvae of this insect possess more than 95% of hemoglobin (Hb) in its circulatory hemolymph. This is a comparative study to see effect of gamma radiation on Hb of Chironomus and humans, two evolutionarily diverse organisms one having extracellular and the other intracellular Hb respectively. Stability and integrity of Chironomus and human Hb to gamma radiation was compared using biophysical techniques like Dynamic Light Scattering (DLS), UV-visible spectroscopy, fluorescence spectrometry and CD spectroscopy after exposure of whole larvae, larval hemolymph, human peripheral blood, purified Chironomus and human Hb. Sequence- and structure-based bioinformatics methods were used to analyze the sequence and structural similarities or differences in the heme pockets of respective Hbs. Resistivity of Chironomus Hb to gamma radiation is remarkably higher than human Hb. Human Hb exhibited loss of heme iron at a relatively low dose of gamma radiation exposure as compared to Chironomus Hb. Unlike human Hb, the heme pocket of Chironomus Hb is rich in aromatic amino acids. Higher hydophobicity around heme pocket confers stability of Chironomus Hb compared to human Hb. Previously reported gamma radiation tolerance of Chironomus can be largely attributed to its evolutionarily ancient form of extracellular Hb as evident from the present study. -- Highlights: •Comparison of radiation tolerant Chironomus Hb and radiation sensitive Human Hb. •Amino acid composition of midge and human heme confer differential hydrophobicity. •Heme pocket of evolutionarily ancient midge Hb provide gamma radiation resistivity.

Potential costs of bacterial infection on storage protein gene expression and reproduction in queenless Apis mellifera worker bees on distinct dietary regimes.

Science.gov (United States)

Lourenço, Anete Pedro; Martins, Juliana Ramos; Guidugli-Lazzarini, Karina Rosa; Macedo, Liliane Maria Fróes; Bitondi, Márcia Maria Gentile; Simões, Zilá Luz Paulino

2012-09-01

Insects are able to combat infection by initiating an efficient immune response that involves synthesizing antimicrobial peptides and a range of other defense molecules. These responses may be costly to the organism, resulting in it exploiting endogenous resources to maintain homeostasis or support defense to the detriment of other physiological needs. We used queenless worker bees on distinct dietary regimes that may alter hemolymph protein storage and ovary activation to investigate the physiological costs of infection with Serratia marcescens. The expression of the genes encoding the storage proteins vitellogenin and hexamerin 70a, the vitellogenin receptor, and vasa (which has a putative role in reproduction), was impaired in the infected bees. This impairment was mainly evident in the bees fed beebread, which caused significantly higher expression of these genes than did royal jelly or syrup, and this was confirmed at the vitellogenin and hexamerin 70a protein levels. Beebread was also the only diet that promoted ovary activation in the queenless bees, but this activation was significantly impaired by the infection. The expression of the genes encoding the storage proteins apolipophorins-I and -III and the lipophorin receptor was not altered by infection regardless the diet provided to the bees. Similarly, the storage of apolipophorin-I in the hemolymph was only slightly impaired by the infection, independently of the supplied diet. Taken together these results indicate that, infection demands a physiological cost from the transcription of specific protein storage-related genes and from the reproductive capacity. Copyright © 2012 Elsevier Ltd. All rights reserved.

Heat Tolerance Induction of the Indian Meal Moth (Lepidoptera: Pyralidae) Is Accompanied by Upregulation of Heat Shock Proteins and Polyols.

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Kim, Minhyun; Lee, Seunghee; Chun, Yong Shik; Na, Jahyun; Kwon, Hyeok; Kim, Wook; Kim, Yonggyun

2017-08-01

The Indian meal moth, Plodia interpunctella, causes massive damage to stored grains and processed foods. Heat treatment has been widely used to control insect pests infesting stored grains. However, heat treatment may result in unsatisfactory control owing to heat tolerance of target insects. This study quantified the heat tolerance and analyzed its induction in P. interpunctella. Susceptibility of P. interpunctella to different high temperatures was assessed in all developmental stages. Heat treatment at 44 °C for 1 h caused significant mortalities to all developmental stages, with late-instar larvae exhibiting the highest tolerance. However, the survivorship to heat treatment was significantly increased by pre-exposure to 37 °C for 30 min. The induction of heat tolerance was accompanied by upregulation of two heat shock proteins of Hsc70 and Hsp90. Trehalose and glycerol concentrations in the hemolymph also increased after pre-exposure to 37 °C for 30 min. RNA interference (RNAi) by specific double-stranded RNAs effectively suppressed the inducible expressions of both Hsc70 and Hsp90 in response to 37 °C for 30 min. Either RNAi of Hsc70 or Hsp90 significantly impaired the heat tolerance induction of P. interpunctella. These results suggest that the induction of heat tolerance in P. interpunctella involves the upregulation of these heat shock proteins and hemolymph polyol levels. © The Authors 2017. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

Differential sensitivity of Chironomus and human hemoglobin to gamma radiation

International Nuclear Information System (INIS)

Gaikwad, Pallavi S.; Panicker, Lata; Mohole, Madhura; Sawant, Sangeeta; Mukhopadhyaya, Rita; Nath, Bimalendu B.

2016-01-01

Chironomus ramosus is known to tolerate high doses of gamma radiation exposure. Larvae of this insect possess more than 95% of hemoglobin (Hb) in its circulatory hemolymph. This is a comparative study to see effect of gamma radiation on Hb of Chironomus and humans, two evolutionarily diverse organisms one having extracellular and the other intracellular Hb respectively. Stability and integrity of Chironomus and human Hb to gamma radiation was compared using biophysical techniques like Dynamic Light Scattering (DLS), UV-visible spectroscopy, fluorescence spectrometry and CD spectroscopy after exposure of whole larvae, larval hemolymph, human peripheral blood, purified Chironomus and human Hb. Sequence- and structure-based bioinformatics methods were used to analyze the sequence and structural similarities or differences in the heme pockets of respective Hbs. Resistivity of Chironomus Hb to gamma radiation is remarkably higher than human Hb. Human Hb exhibited loss of heme iron at a relatively low dose of gamma radiation exposure as compared to Chironomus Hb. Unlike human Hb, the heme pocket of Chironomus Hb is rich in aromatic amino acids. Higher hydophobicity around heme pocket confers stability of Chironomus Hb compared to human Hb. Previously reported gamma radiation tolerance of Chironomus can be largely attributed to its evolutionarily ancient form of extracellular Hb as evident from the present study. -- Highlights: •Comparison of radiation tolerant Chironomus Hb and radiation sensitive Human Hb. •Amino acid composition of midge and human heme confer differential hydrophobicity. •Heme pocket of evolutionarily ancient midge Hb provide gamma radiation resistivity.

Ca2+-dependent proteolytic activity in crab claw muscle: effects of inhibitors and specificity for myofibrillar proteins

International Nuclear Information System (INIS)

Mykles, D.L.; Skinner, D.M.

1983-01-01

The claw closer muscle of the Bermuda land crab, Gecarcinus lateralis, undergoes a sequential atrophy and restoration during each molting cycle. The role of Ca 2+ -dependent proteinases in the turn-over of myofibrillar protein in normal anecdysial (intermolt) claw muscle is described. Crab Ca 2+ -dependent proteinase degrades the myofibrillar proteins actin, myosin heavy and light chains, paramyosin, tropomyosin, and troponin-T and -I. Ca 2+ -dependent proteinase activity in whole homogenates and 90,000 x g supernatant fractions from muscle homogenates has been characterized with respect to Ca 2+ requirement, substrate specificity, and effects of proteinase inhibitors. The enzyme is inhibited by antipain, leupeptin, E-64, and iodoacetamide; it is insensitive to pepstatin A. The specificity of crab Ca 2+ -dependent proteinase was examined with native myosin with normal ATPase activity as well as with radioiodinated myosin and radioiodinated hemolymph proteins. Hydrolysis of 125 I-myosin occurs in two phases, both Ca 2+ -dependent: (1) heavy chain (M/sub r/ = 200,000) is cleaved into four large fragments (M/sub r/ = 160,000, 110,000, 73,000, 60,000) and numerous smaller fragments; light chain (M/sub r/ = 18,000) is cleaved to a 15,000-Da fragment; (2) the fragments produced in the first phase are hydrolyzed to acid-soluble material. Although radioiodinated native hemolymph proteins are not susceptible to the Ca 2+ -dependent proteinase, those denatured by carboxymethylation are degraded. These data suggest that crab Ca 2+ -dependent proteinase is involved in turnover of myofibrillar protein in normal muscle and muscle undergoing proecdysial atrophy

Antibacterial activity of hemolymph of mud crab, Scylla serrata to marine and freshwater microbes

Digital Repository Service at National Institute of Oceanography (India)

Sabu, A.S.; Chatterji, A.

stream_size 9 stream_content_type text/plain stream_name Diversity_Life_Process_Ocean_Land_2007_14.pdf.txt stream_source_info Diversity_Life_Process_Ocean_Land_2007_14.pdf.txt Content-Encoding ISO-8859-1 Content-Type text...

[Radioecological studies of freshwater mollusks in the Chernobyl accident exclusion zone].

Science.gov (United States)

Gudkov, D I; Nazarov, A B; Dziubenko, E V; Kaglian, A E; Klenus, V G

2009-01-01

Species-specificity and dynamics of 90Sr, 137Cs and some transuranic elements accumulation in bivalve and gastropod freshwater molluscs of the Chernobyl exclusion zone during 1997-2008 was analyzed. The results of radiation dose and chromosome aberration rate estimation and the analysis of hemolymph composition of freshwater snail (Lymnaea stagnalis L.) was produced. The absorbed dose rate was registered in the range of 0.3-85.0 microGy/h. In closed water bodies the heightened chromosome aberration rate (up to 27%) in embryo tissues, and also the change of haematological indexes for the adult individuals of snails was registered.

The entomopathogenic fungus Nomuraea rileyi impairs cellular immunity of its host Helicoverpa armigera.

Science.gov (United States)

Zhong, Ke; Liu, Zhan-Chi; Wang, Jia-Lin; Liu, Xu-Sheng

2017-09-01

In this study, we investigated the effect of the entomopathogenic fungus Nomuraea rileyi on Helicoverpa armigera cellular immune responses. Nomuraea rileyi infection had no effect on total hemocyte count (THC), but impaired hemocyte-mediated phagocytosis, nodulation, and encapsulation responses. Nomuraea rileyi infection led to a significant reduction in hemocyte spreading. An in vitro assay revealed that plasma from N. rileyi infected H. armigera larvae suppressed the spreading ability of hemocytes from naïve larvae. We infer that N. rileyi suppresses the cellular immune response of its host, possibly by secreting exogenous, cytotoxic compounds into the host's hemolymph. © 2017 Wiley Periodicals, Inc.

Antioxidant properties of species from the Brazilian cerrado by different assays

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K.S. Farias

2013-01-01

Full Text Available The purpose of this study was to screen the antioxidant activity of medicinal plant extracts from the Brazilian cerrado, through other methods than the total phenolic content and its correlation with the antioxidant activity. Ethanolic extracts of ten species were evaluated through three antioxidant assays, in vitro, including 2,2-diphenyl-1-picrylhydrazyl (DPPH, total antioxidant activity and reducing power; and by using the Folin-Ciocalteu method the total phenolic content was determined. Ethanolic extracts of Stryphnodendron obovatum, Cecropia pachystachya and Duguetia furfuraceae showed strong antioxidant activity (IC50<5 µg mL-1 in the DPPH free radical scavenging assay; the species Vernonia phosphorea, Hymenaea stignocarpa and Jacaranda ulei may also be highlighted. These results were confirmed in the assays of total antioxidant capacity and reducing power. The extracts of S. obovatum and V. phosphorea showed an abundant phenolic content; therefore, the phenolic content may play a role in the antioxidant activity. These two species, traditionally used in Brazil, showed great power in these assay systems and may be a promising source for the development of natural antioxidants and future candidates for phytochemical and pharmacological studies in related diseases.

Medicinal and ethnoveterinary remedies of hunters in Trinidad

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Georges Karla

2001-11-01

Full Text Available Abstract Background Ethnomedicines are used by hunters for themselves and their hunting dogs in Trinidad. Plants are used for snakebites, scorpion stings, for injuries and mange of dogs and to facilitate hunting success. Results Plants used include Piper hispidum, Pithecelobium unguis-cati, Bauhinia excisa, Bauhinia cumanensis, Cecropia peltata, Aframomum melegueta, Aristolochia rugosa, Aristolochia trilobata, Jatropha curcas, Jatropha gossypifolia, Nicotiana tabacum, Vernonia scorpioides, Petiveria alliacea, Renealmia alpinia, Justicia secunda, Phyllanthus urinaria,Phyllanthus niruri,Momordica charantia, Xiphidium caeruleum, Ottonia ovata, Lepianthes peltata, Capsicum frutescens, Costus scaber, Dendropanax arboreus, Siparuma guianensis, Syngonium podophyllum, Monstera dubia, Solanum species, Eclipta prostrata, Spiranthes acaulis, Croton gossypifolius, Barleria lupulina, Cola nitida, Acrocomia ierensis (tentative ID. Conclusion Plant use is based on odour, and plant morphological characteristics and is embedded in a complex cultural context based on indigenous Amerindian beliefs. It is suggested that the medicinal plants exerted a physiological action on the hunter or his dog. Some of the plants mentioned contain chemicals that may explain the ethnomedicinal and ethnoveterinary use. For instance some of the plants influence the immune system or are effective against internal and external parasites. Plant baths may contribute to the health and well being of the hunting dogs.

Vascular epiphytes as regeneration indicators of disturbed forests of the Colombian Amazon region

International Nuclear Information System (INIS)

Triana Moreno, Luz Amparo; Garzon Venegas, Nelson Javier; Sanchez Zambrano, Jairo; Vargas Orlando

2003-01-01

In order to compare how the distribution and composition of vascular epiphytes varies, in three disturbed forests with different recovery times, and to verify whether these factors can indicate the regeneration state, three stubbles that had been abandoned during 12, 18 and 22 years were selected in the neighborhood of Leticia City (Amazons, Colombia). In each stubble 7 Cecropia sciadophylla (Cecropiaceae) individuals were selected, and a sampling of epiphytes was made in the first 3 m of each tree. The number of species, their abundance and covering were used as criteria to compare the three stubbles, because the sensitivity of epiphytes to environmental changes. The results show that the evaluated factors are useful for the characterization of the forest regeneration process. In spite of the fact that the number of species in the there stubbles was similar, the composition varied in such ways that about half the species of each stubble, were exclusive. The dominance of Monstera obliqua (Araceae) was evident, constituting more than 80% of the epiphytic covering of the total sample. The youngest stubble presented a denser covering and a high diversity index, whereas in the oldest stubbles these values diminish drastically

Predation and aggressiveness in host plant protection: a generalization using ants from the genus Azteca

Science.gov (United States)

Dejean, Alain; Grangier, Julien; Leroy, Céline; Orivel, Jerôme

2009-01-01

In studying the ant genus Azteca, a Neotropical group of arboreal species, we aimed to determine the extent to which the ants use predation and/or aggressiveness to protect their host plants from defoliating insects. We compared a territorially dominant, carton-nester, Azteca chartifex, and three plant-ant species. Azteca alfari and Azteca ovaticeps are associated with the myrmecophyte Cecropia (Cecropiaceae) and their colonies shelter in its hollow branches; whereas Azteca bequaerti is associated with Tococa guianensis (Melastomataceae) and its colonies shelter in leaf pouches situated at the base of the laminas. Whereas A. bequaerti workers react to the vibrations transmitted by the lamina when an alien insect lands on a leaf making it unnecessary for them to patrol their plant, the workers of the three other species rather discover prey by contact. The workers of all four species use a predatory behaviour involving spread-eagling alien insects after recruiting nestmates at short range, and, in some cases, at long range. Because A. alfari and A. ovaticeps discard part of the insects they kill, we deduced that the workers’ predatory behaviour and territorial aggressiveness combine in the biotic defence of their host tree.

Medicinal and ethnoveterinary remedies of hunters in Trinidad.

Science.gov (United States)

Lans, C; Harper, T; Georges, K; Bridgewater, E

2001-01-01

Ethnomedicines are used by hunters for themselves and their hunting dogs in Trinidad. Plants are used for snakebites, scorpion stings, for injuries and mange of dogs and to facilitate hunting success. Plants used include Piper hispidum, Pithecelobium unguis-cati, Bauhinia excisa, Bauhinia cumanensis, Cecropia peltata, Aframomum melegueta, Aristolochia rugosa, Aristolochia trilobata, Jatropha curcas, Jatropha gossypifolia, Nicotiana tabacum, Vernonia scorpioides, Petiveria alliacea, Renealmia alpinia, Justicia secunda, Phyllanthus urinaria,Phyllanthus niruri,Momordica charantia, Xiphidium caeruleum, Ottonia ovata, Lepianthes peltata, Capsicum frutescens, Costus scaber, Dendropanax arboreus, Siparuma guianensis, Syngonium podophyllum, Monstera dubia, Solanum species, Eclipta prostrata, Spiranthes acaulis, Croton gossypifolius, Barleria lupulina, Cola nitida, Acrocomia ierensis (tentative ID). Plant use is based on odour, and plant morphological characteristics and is embedded in a complex cultural context based on indigenous Amerindian beliefs. It is suggested that the medicinal plants exerted a physiological action on the hunter or his dog. Some of the plants mentioned contain chemicals that may explain the ethnomedicinal and ethnoveterinary use. For instance some of the plants influence the immune system or are effective against internal and external parasites. Plant baths may contribute to the health and well being of the hunting dogs.

Estabelecimento de Espécies Florestais Nativas Via Semeadura Direta no Rio Piauitinga - Sergipe

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Janisson Batista de Jesus

2017-04-01

Full Text Available RESUMO A técnica de semeadura direta vem se destacando nos projetos de recuperação florestal pela redução de custos na implantação e tem sido cada vez mais utilizada. Este trabalho foi realizado com o objetivo de avaliar a emergência, sobrevivência e estabelecimento de espécies florestais nativas, por meio de sementes em áreas de mata ciliar no Rio Piauitinga, município de Lagarto, SE. As espécies utilizadas foram Libidibia ferrea var. leiostachya (Benth. L. P. Queiroz, Cassia grandis L. f., Cecropia pachystachya Trec., Enterolobium contortisiliquum (Vell. Morong e Guazuma ulmifolia Lam. O experimento em campo foi implantado em Delineamento em Blocos Casualizados (DBC com quatro repetições, em esquema fatorial, testando-se as cinco espécies com e sem tratamento para superação de dormência. A semeadura direta mostrou-se uma técnica viável para a área de estudo apenas para as espécies L. ferrea var. leiostachya, E. contortisiliquum e Cassia grandis, utilizando-se sementes com e sem dormência.

Estrutura fitossociológica do componente arbóreo de um fragmento de Floresta Ombrófila Densa na mata sul de Pernambuco, nordeste do Brasil

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Roberto Felix Costa

2008-01-01

Full Text Available This work aimed to describe physiognomy and structure of a Dense Ombrophyllous Forest fragment located in Catende, Pernambuco. All trees with DAP (breast height diameter ≥ 4.77 cm were measured in 40 10 x 25 m plots, systematically installed in five transects. A total of 1049 individuals were observed, distributed in 91 species, 64 genera and 37 botanical families. Mimosaceae and Lauraceae were the families with higher species number, while Anacardiaceae and Moraceae had the highest individuals numbers. Shannon index (H´ was 3.83 nats/ind. Total basal area, height and diameter (average and maximum were 23.59 m²/ha, 13.57 m (± 7.13, 45 m, 13.20 cm (± 10.60 and 127.32 cm, respectively. The species with higher importance values were Tapirira guianensis Aubl., Dialium guianense (Aubl. Sandwith, Helicostylis tomentosa (Poepp. & Endl. Rusby, Thyrsodium spruceanum Benth., Plathymenia foliolosa Benth., Brosimum discolor Schott, Parkia pendula (Willd. Benth. ex Walp., Schefflera morototoni (Aubl. Maguire, Steyerm. & Frodin, Eschweilera ovata (Cambess. Miers and Cecropia palmata Willd. Diametric distribution had an inverted-J-like curve, predicted for a forest unequal in ages in a secondary succession stage.

The Postpharyngeal Gland: Specialized Organ for Lipid Nutrition in Leaf-Cutting Ants.

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Decio, Pâmela; Vieira, Alexsandro Santana; Dias, Nathalia Baptista; Palma, Mario Sergio; Bueno, Odair Correa

2016-01-01

There are several hypotheses about the possible functions of the postpharyngeal gland (PPG) in ants. The proposed functions include roles as cephalic or gastric caeca and diverticulum of the digestive tract, mixing of hydrocarbons, nestmate recognition, feeding larvae, and the accumulation of lipids inside this gland, whose origin is contradictory. The current study aimed to investigate the functions of these glands by examining the protein expression profile of the PPGs of Atta sexdens rubropilosa (Hymenoptera, Formicidae). Mated females received lipid supplementation and their glands were extracted and analyzed using a proteomic approach. The protocol used combined two-dimensional electrophoresis and shotgun strategies, followed by mass spectrometry. We also detected lipid β-oxidation by immunofluorescent marking of acyl-CoA dehydrogenase. Supplying ants with lipids elicited responses in the glandular cells of the PPG; these included increased expression of proteins related to defense mechanisms and signal transduction and reorganization of the cytoskeleton due to cell expansion. In addition, some proteins in PPG were overexpressed, especially those involved in lipid and energy metabolism. Part of the lipids may be reduced, used for the synthesis of fatty alcohol, transported to the hemolymph, or may be used as substrate for the synthesis of acetyl-CoA, which is oxidized to form molecules that drive oxidative phosphorylation and produce energy for cellular metabolic processes. These findings suggest that this organ is specialized for lipid nutrition of adult leaf-cutting ants and characterized like a of diverticulum foregut, with the ability to absorb, store, metabolize, and mobilize lipids to the hemolymph. However, we do not rule out that the PPG may have other functions in other species of ants.

The Role of DmCatD, a Cathepsin D-Like Peptidase, and Acid Phosphatase in the Process of Follicular Atresia in Dipetalogaster maxima (Hemiptera: Reduviidae), a Vector of Chagas' Disease

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Leyria, Jimena; Fruttero, Leonardo L.; Nazar, Magalí; Canavoso, Lilián E.

2015-01-01

In this work, we have investigated the involvement of DmCatD, a cathepsin D-like peptidase, and acid phosphatase in the process of follicular atresia of Dipetalogaster maxima, a hematophagous insect vector of Chagas’ disease. For the studies, fat bodies, ovaries and hemolymph were sampled from anautogenous females at representative days of the reproductive cycle: pre-vitellogenesis, vitellogenesis as well as early and late atresia. Real time PCR (qPCR) and western blot assays showed that DmCatD was expressed in fat bodies and ovaries at all reproductive stages, being the expression of its active form significantly higher at the atretic stages. In hemolymph samples, only the immunoreactive band compatible with pro-DmCatD was observed by western blot. Acid phosphatase activity in ovarian tissues significantly increased during follicular atresia in comparison to pre-vitellogenesis and vitellogenesis. A further enzyme characterization with inhibitors showed that the high levels of acid phosphatase activity in atretic ovaries corresponded mainly to a tyrosine phosphatase. Immunofluorescence assays demonstrated that DmCatD and tyrosine phosphatase were associated with yolk bodies in vitellogenic follicles, while in atretic stages they displayed a different cellular distribution. DmCatD and tyrosine phosphatase partially co-localized with vitellin. Moreover, their interaction was supported by FRET analysis. In vitro assays using homogenates of atretic ovaries as the enzyme source and enzyme inhibitors demonstrated that DmCatD, together with a tyrosine phosphatase, were necessary to promote the degradation of vitellin. Taken together, the results strongly suggested that both acid hydrolases play a central role in early vitellin proteolysis during the process of follicular atresia. PMID:26091289

OpnS, an Outer Membrane Porin of Xenorhabdus nematophila, Confers a Competitive Advantage for Growth in the Insect Host▿ †

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van der Hoeven, Ransome; Forst, Steven

2009-01-01

The gammaproteobacterium Xenorhabdus nematophila engages in a mutualistic association with an entomopathogenic nematode and also functions as a pathogen toward different insect hosts. We studied the role of the growth-phase-regulated outer membrane protein OpnS in host interactions. OpnS was shown to be a 16-stranded β-barrel porin. opnS was expressed during growth in insect hemolymph and expression was elevated as the cell density increased. When wild-type and opnS deletion strains were coinjected into insects, the wild-type strain was predominantly recovered from the insect cadaver. Similarly, an opnS-complemented strain outcompeted the ΔopnS strain. Coinjection of the wild-type and ΔopnS strains together with uncolonized nematodes into insects resulted in nematode progeny that were almost exclusively colonized with the wild-type strain. Likewise, nematode progeny recovered after coinjection of a mixture of nematodes carrying either the wild-type or ΔopnS strain were colonized by the wild-type strain. In addition, the ΔopnS strain displayed a competitive growth defect when grown together with the wild-type strain in insect hemolymph but not in defined culture medium. The ΔopnS strain displayed increased sensitivity to antimicrobial compounds, suggesting that deletion of OpnS affected the integrity of the outer membrane. These findings show that the OpnS porin confers a competitive advantage for the growth and/or the survival of X. nematophila in the insect host and provides a new model for studying the biological relevance of differential regulation of porins in a natural host environment. PMID:19465651

Lactobacillus paracasei modulates the immune system of Galleria mellonella and protects against Candida albicans infection

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Rossoni, Rodnei Dennis; Fuchs, Beth Burgwyn; de Barros, Patrícia Pimentel; Velloso, Marisol dos Santos; Jorge, Antonio Olavo Cardoso; Junqueira, Juliana Campos; Mylonakis, Eleftherios

2017-01-01

Probiotics have been described as a potential strategy to control opportunistic infections due to their ability to stimulate the immune system. Using the non-vertebrate model host Galleria mellonella, we evaluated whether clinical isolates of Lactobacillus spp. are able to provide protection against Candida albicans infection. Among different strains of Lactobacillus paracasei, Lactobacillus rhamnosus and Lactobacillus fermentum, we verified that L. paracasei 28.4 strain had the greatest ability to prolong the survival of larvae infected with a lethal dose of C. albicans. We found that the injection of 107 cells/larvae of L. paracasei into G. mellonella larvae infected by C. albicans increased the survival of these insects compared to the control group (P = 0.0001). After that, we investigated the immune mechanisms involved in the protection against C. albicans infection, evaluating the number of hemocytes and the gene expression of antifungal peptides. We found that L. paracasei increased the hemocyte quantity (2.38 x 106 cells/mL) in relation to the control group (1.29 x 106 cells/mL), indicating that this strain is capable of raising the number of circulating hemocytes into the G. mellonella hemolymph. Further, we found that L. paracasei 28.4 upregulated genes that encode the antifungal peptides galiomicin and gallerymicin. In relation to the control group, L. paracasei 28.4 increased gene expression of galiomicin by 6.67-fold and 17.29-fold for gallerymicin. Finally, we verified that the prophylactic provision of probiotic led to a significant reduction of the number of fungal cells in G. mellonella hemolymph. In conclusion, L. paracasei 28.4 can modulate the immune system of G. mellonella and protect against candidiasis. PMID:28267809

Lactobacillus paracasei modulates the immune system of Galleria mellonella and protects against Candida albicans infection.

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Rossoni, Rodnei Dennis; Fuchs, Beth Burgwyn; de Barros, Patrícia Pimentel; Velloso, Marisol Dos Santos; Jorge, Antonio Olavo Cardoso; Junqueira, Juliana Campos; Mylonakis, Eleftherios

2017-01-01

Probiotics have been described as a potential strategy to control opportunistic infections due to their ability to stimulate the immune system. Using the non-vertebrate model host Galleria mellonella, we evaluated whether clinical isolates of Lactobacillus spp. are able to provide protection against Candida albicans infection. Among different strains of Lactobacillus paracasei, Lactobacillus rhamnosus and Lactobacillus fermentum, we verified that L. paracasei 28.4 strain had the greatest ability to prolong the survival of larvae infected with a lethal dose of C. albicans. We found that the injection of 107 cells/larvae of L. paracasei into G. mellonella larvae infected by C. albicans increased the survival of these insects compared to the control group (P = 0.0001). After that, we investigated the immune mechanisms involved in the protection against C. albicans infection, evaluating the number of hemocytes and the gene expression of antifungal peptides. We found that L. paracasei increased the hemocyte quantity (2.38 x 106 cells/mL) in relation to the control group (1.29 x 106 cells/mL), indicating that this strain is capable of raising the number of circulating hemocytes into the G. mellonella hemolymph. Further, we found that L. paracasei 28.4 upregulated genes that encode the antifungal peptides galiomicin and gallerymicin. In relation to the control group, L. paracasei 28.4 increased gene expression of galiomicin by 6.67-fold and 17.29-fold for gallerymicin. Finally, we verified that the prophylactic provision of probiotic led to a significant reduction of the number of fungal cells in G. mellonella hemolymph. In conclusion, L. paracasei 28.4 can modulate the immune system of G. mellonella and protect against candidiasis.

Efficacy of Aloe Vera Cream in the Treatment of Paederus Dermatitis in Mice

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Ramin Khaghani

2017-06-01

Full Text Available Background: Dermatitis caused by Paederus beetle involves many people around the world, especially Iran. The symptoms include redness, itching and severe irritation. This study evaluated the effectiveness of the Aloe vera cream on the treatment of dermatitis caused by Paederus beetles.Methods: Forty male 6–8 weeks BALB/C mice were randomly divided into four groups of 10 mice. After removing the mice’s back hair, the backs of mice were marked by a circle with a diameter of 3 mm. The Paederus beetles were collected from Babol in Mazandaran Province, northern Iran and transferred to the animal lab of Aja University of Medical Sciences, Tehran, Iran. The end of abdominal segment Paederus was cut with scissors and hemolymph content was pushed by forceps on the circle. Only hemolymph of one Paederus applied to the back of each mouse. Groups 1, 2, 3 and 4 were treated with the base (vehicle, dexamethasone 0.1%, Aloe vera 0.5% and Aloe vera 2% creams respectively. After 2 days, dermatitis appeared. Then the mentioned creams were applied on the mice once a day. The wound area was measured every day. Dermatitis surface area under curve (AUC of each mouse was cal­culated for 17 days after induction of dermatitis. Statistical analysis of ANOVA was used.Results: Application of Aloe vera 0.5% and 2% significantly reduced the healing duration and dermatitis area in comparison with the vehicle and dexamethasone cream (P< 0.05. But dexamethasone had no significant effect on the healing of dermatitis as compared to vehicle.Conclusion: Aloe vera may clinically effective in the treatment of Paederus dermatitis.

A novel member of the trehalose transporter family functions as an H+-dependent trehalose transporter in the reabsorption of trehalose in Malpighian tubules.

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Shingo eKikuta

2012-07-01

Full Text Available In insects, Malpighian tubules are functionally analogous to mammalian kidneys in that they not only are essential to excrete waste molecules into the lumen but also are responsible for the reabsorption of indispensable molecules, such as sugars, from the lumen to the principal cells. Among sugars, the disaccharide trehalose is highly important to insects because it is the main hemolymph sugar to serve as a source of energy and carbon. The trehalose transporter TRET1 participates in the transfer of newly synthesized trehalose from the fat body across the cellular membrane into the hemolymph. Although transport proteins must play a pivotal role in the reabsorption of trehalose in Malpighian tubules, the molecular context underlying this process remains obscure. Previously, we identified a Tret1 homolog (Nlst8 that is expressed principally in the Malpighian tubules of the brown planthopper (BPH. Here, we used the Xenopus oocyte expression system to show that NlST8 exerts trehalose transport activity that is elevated under low pH conditions. These functional assays indicate that Nlst8 encodes a proton-dependent trehalose transporter (H-TRET1. To examine the involvement of Nlst8 in trehalose reabsorption, we analyzed the sugar composition of honeydew by using BPH with RNAi gene silencing. Trehalose was detected in the honeydew as waste excreted from Nlst8-dsRNA-injected BPH under hyperglycemic conditions. However, trehalose was not expelled from GFP-dsRNA-injected BPH even under hyperglycemic conditions. We conclude that NlST8 could participate in trehalose reabsorption driven by a H+ gradient from the lumen to the principal cells of the Malpighian tubules.

No genetic tradeoffs between hygienic behaviour and individual innate immunity in the honey bee, Apis mellifera.

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Harpur, Brock A; Chernyshova, Anna; Soltani, Arash; Tsvetkov, Nadejda; Mahjoorighasrodashti, Mohammad; Xu, Zhixing; Zayed, Amro

2014-01-01

Many animals have individual and social mechanisms for combating pathogens. Animals may exhibit short-term physiological tradeoffs between social and individual immunity because the latter is often energetically costly. Genetic tradeoffs between these two traits can also occur if mutations that enhance social immunity diminish individual immunity, or vice versa. Physiological tradeoffs between individual and social immunity have been previously documented in insects, but there has been no study of genetic tradeoffs involving these traits. There is strong evidence that some genes influence both innate immunity and behaviour in social insects--a prerequisite for genetic tradeoffs. Quantifying genetic tradeoffs is critical for understanding the evolution of immunity in social insects and for devising effective strategies for breeding disease-resistant pollinator populations. We conducted two experiments to test the hypothesis of a genetic tradeoff between social and individual immunity in the honey bee, Apis mellifera. First, we estimated the relative contribution of genetics to individual variation in innate immunity of honey bee workers, as only heritable traits can experience genetic tradeoffs. Second, we examined if worker bees with hygienic sisters have reduced individual innate immune response. We genotyped several hundred workers from two colonies and found that patriline genotype does not significantly influence the antimicrobial activity of a worker's hemolymph. Further, we did not find a negative correlation between hygienic behaviour and the average antimicrobial activity of a worker's hemolymph across 30 honey bee colonies. Taken together, our work indicates no genetic tradeoffs between hygienic behaviour and innate immunity in honey bees. Our work suggests that using artificial selection to increase hygienic behaviour of honey bee colonies is not expected to concurrently compromise individual innate immunity of worker bees.

The specificity of immune priming in silkworm, Bombyx mori, is mediated by the phagocytic ability of granular cells.

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Wu, Gongqing; Li, Mei; Liu, Yi; Ding, Ying; Yi, Yunhong

2015-10-01

In the past decade, the phenomenon of immune priming was documented in many invertebrates in a large number of studies; however, in most of these studies, behavioral evidence was used to identify the immune priming. The underlying mechanism and the degree of specificity of the priming response remain unclear. We studied the mechanism of immune priming in the larvae of the silkworm, Bombyx mori, and analyzed the specificity of the priming response using two closely related Gram-negative pathogenic bacteria (Photorhabdus luminescens TT01 and P. luminescens H06) and one Gram-positive pathogenic bacterium (Bacillus thuringiensis HD-1). Primed with heat-killed bacteria, the B. mori larvae were more likely to survive subsequent homologous exposure (the identical bacteria used in the priming and in the subsequent challenge) than heterologous (different bacteria used in the priming and subsequent exposure) exposure to live bacteria. This result indicated that the B. mori larvae possessed a strong immune priming response and revealed a degree of specificity to TT01, H06 and HD-1 bacteria. The degree of enhanced immune protection was positively correlated with the level of phagocytic ability of the granular cells and the antibacterial activity of the cell-free hemolymph. Moreover, the granular cells of the immune-primed larvae increased the phagocytosis of a previously encountered bacterial strain compared with other bacteria. Thus, the enhanced immune protection of the B. mori larvae after priming was mediated by the phagocytic ability of the granular cells and the antibacterial activity of the hemolymph; the specificity of the priming response was primarily attributed to the phagocytosis of bacteria by the granular cells. Copyright © 2015 Elsevier Ltd. All rights reserved.

Experimental infection in Cavia porcellus by infected Amblyomma ovale nymphs with Rickettsia sp. (Atlantic rainforest strain).

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Brustolin, Joice Magali; da Silva Krawczak, Felipe; Alves, Marta Elena Machado; Weiller, Maria Amélia; de Souza, Camila Lopes; Rosa, Fábio Brum; Cadore, Gustavo Cauduro; Dos Anjos Lopes, Sônia Terezinha; Labruna, Marcelo Bahia; Vogel, Fernanda Silveira Flores; de Avila Botton, Sônia; Sangioni, Luís Antônio

2018-03-01

This study describes experimental infection of guinea pigs (Cavia porcellus) infested with naturally infected Amblyomma ovale nymphs with Rickettsia sp. (Atlantic rainforest strain), and the capacity of A. ovale nymphs to transmit this bacterium. Twenty-six guinea pigs were divided into the following groups: G1, 10 animals infested with uninfected A. ovale nymphs; G2, 10 animals infested with nymphs infected with Rickettsia sp. (Atlantic rainforest strain); and G3, 6 animals without tick infestation. Blood samples were taken 7, 14, 21, and 28 days post-infestation for serological and hematological tests. For histopathological analysis and rickettsial DNA detection, fragments of the spleen, lung, brain, and liver were harvested after euthanasia. The average feeding period for nymphs was 6.6 days for G1 and 6 days for G2. Hemolymph and PCR assays, performed to detect the causative agent in ticks, indicated that in G1, all ticks were negative, and in G2, all nymphs were positive by PCR and 80% (8/10) was positive by hemolymph tests. The only clinical change was skin scarring at the tick attachment site. Hematological parameters indicated leukopenia and total plasma protein (TPP) increased with decreased platelets in G1. In G2, leukocytosis, neutrophilia, monocytosis, an increase in platelets, and reduced TPP were observed. Only G2 guinea pigs were seroconverted (80%; 8/10). Histopathology tests indicated mild, diffuse hemosiderosis and mild, multifocal, follicular hyperplasia in the spleen. Molecular analysis did not detect Rickettsia sp. DNA in C. porcellus tissues. We demonstrated the capacity of A. ovale nymphs to transmit Rickettsia sp. (Atlantic rainforest strain) to guinea pigs.

High-resolution mass spectrometry driven discovery of peptidic danger signals in insect immunity.

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Arton Berisha

Full Text Available The 'danger model' is an alternative concept for immune response postulating that the immune system reacts to entities that do damage (danger associated molecular patterns, DAMP and not only to entities that are foreign (pathogen-associated molecular patterns, PAMP as proposed by classical immunology concepts. In this study we used Galleria mellonella to validate the danger model in insects. Hemolymph of G. mellonella was digested with thermolysin (as a representative for virulence-associated metalloproteinases produced by humanpathogens followed by chromatographic fractionation. Immune-stimulatory activity was tested by measuring lysozyme activity with the lytic zone assays against Micrococcus luteus cell wall components. Peptides were analyzed by nano-scale liquid chromatography coupled to high-resolution Fourier transform mass spectrometers. Addressing the lack of a genome sequence we complemented the rudimentary NCBI protein database with a recently established transcriptome and de novo sequencing methods for peptide identification. This approach led to identification of 127 peptides, 9 of which were identified in bioactive fractions. Detailed MS/MS experiments in comparison with synthetic analogues confirmed the amino acid sequence of all 9 peptides. To test the potential of these putative danger signals to induce immune responses we injected the synthetic analogues into G. mellonella and monitored the anti-bacterial activity against living Micrococcus luteus. Six out of 9 peptides identified in the bioactive fractions exhibited immune-stimulatory activity when injected. Hence, we provide evidence that small peptides resulting from thermolysin-mediated digestion of hemolymph proteins function as endogenous danger signals which can set the immune system into alarm. Consequently, our study indicates that the danger model also plays a role in insect immunity.

Identification of a female spawn-associated Kazal-type inhibitor from the tropical abalone Haliotis asinina.

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Wang, Tianfang; Nuurai, Parinyaporn; McDougall, Carmel; York, Patrick S; Bose, Utpal; Degnan, Bernard M; Cummins, Scott F

2016-07-01

Abalone (Haliotis) undergoes a period of reproductive maturation, followed by the synchronous release of gametes, called broadcast spawning. Field and laboratory studies have shown that the tropical species Haliotis asinina undergoes a two-week spawning cycle, thus providing an excellent opportunity to investigate the presence of endogenous spawning-associated peptides. In female H. asinina, we have isolated a peptide (5145 Da) whose relative abundance in hemolymph increases substantially just prior to spawning and is still detected using reverse-phase high-performance liquid chromatography chromatograms up to 1-day post-spawn. We have isolated this peptide from female hemolymph as well as samples prepared from the gravid female gonad, and demonstrated through comparative sequence analysis that it contains features characteristic of Kazal-type proteinase inhibitors (KPIs). Has-KPI is expressed specifically within the gonad of adult females. A recombinant Has-KPI was generated using a yeast expression system. The recombinant Has-KPI does not induce premature spawning of female H. asinina when administered intramuscularly. However it displays homomeric aggregations and interaction with at least one mollusc-type neuropeptide (LRDFVamide), suggesting a role for it in regulating neuropeptide endocrine communication. This research provides new understanding of a peptide that can regulate reproductive processes in female abalone, which has the potential to lead to the development of greater control over abalone spawning. The findings also highlight the need to further explore abalone reproduction to clearly define a role for novel spawning-associated peptide in sexual maturation and spawning. Copyright © 2016 European Peptide Society and John Wiley & Sons, Ltd. Copyright © 2016 European Peptide Society and John Wiley & Sons, Ltd.

Protein content and electrophoretic profile of fat body and ovary extracts from workers of Melipona quadrifasciata anthidioides (Hymenoptera, Meliponini

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Vagner T. Paes de Oliveira

Full Text Available Workers of Melipona quadrifasciata anthidioides (Lepeletier, 1836 develop their ovaries and lay eggs, therefore the production of vitellogenin is expected. In electrophoretic profiles only fat body extracts from nurse workers and ovary extracts from newly-emerged workers show protein with molecular mass similar to vitellogenin. However, an increase in the protein content was detected in forager fat body. This increase was attributed to storage of vitellogenin or other proteins in the previous phase and not discharged into the hemolymph or to an effect of the increased titre of juvenile hormone in this phase of worker life over the fat body functioning.

Influence of the Chernobyl NPP accident consequences to the animal kingdom. Chapter 4

International Nuclear Information System (INIS)

Pikulik, M.M.; Plenin, A.E.; Galkovskaya, G.A.; Zarubov, A.I.; Molotkov, D.V.; Khmeleva, N.N.; Golubev, A.P.; Moroz, M.D.; Petukhov, V.B.; Petrikov, A.M.; Kokhnenko, O.S.; Shevtsova, T.M.; Ermolaev, V.V.; Ermolaeva, I.A.; Maksimova, S.L.; Matveenko, A.A.; Blinov, V.V.; Shlyakhtenok, A.S.; Eliseeva, K.G.; Vojtovich, A.M.; Trusova, V.D.; Ogurtsova, S.Eh.; Drobenkov, S.M.; Nikiforov, M.E.; Tishechkin, A.K.; Samusenko, I.Eh.; Parejko, O.A.; Goncharova, R.I.; Ryabokon', N.I.; Rozhdestvenskaya, A.S.; Sidorovich, V.E.; Kozlo, P.G.; Dunin, V.F.; Kuchmel', S.V.; Deryabina, T.G.

1995-01-01

The total regularities of ionizing irradiation effect on the living systems, as well as molecular, cell and tissues radiation effects, radiation effects on organism, population and ecosystem are shown. The results of monitoring of zoo plankton systems, state of populations of mass species of benthos invertebrates and fishes in water reservoirs in the Chernobyl accident zone are given. Disorders in hemolymph and changes in complexes of soil invertebrates, as well as changes of population structures of amphibians and reptiles living in radioactive contaminated zone are described. The state of model groups and forming of population structure of insects, birds and mammals were investigated. 19 tabs., 16 figs

Estrutura fitossociológia de um fragmento natural de floresta inundável em área de orizicultura irrigada, município de Lagoa da Confusão, Tocantins Phytosociologial structure of a natural fragment of floodplain forest in area of irrigated rice cultivation, municipal district of Lagoa da Confusão, Tocantins, Brazil

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Elizabeth Rodrigues Brito

2006-10-01

Full Text Available Os fragmentos naturais de florestas inundáveis conhecidos como ipucas localizam-se na planície do Araguaia, sob a forma de depressões naturais, que no Estado do Tocantins estão situados em áreas de planícies de inundação, que favorece seu alagamento e, conseqüentemente, o maior tempo de retenção da água em épocas de elevada precipitação. O presente estudo foi desenvolvido na fazenda Lago Verde, Município de Lagoa da Confusão, Tocantins, situado entre as coordenadas UTM: 643586 e 644060 leste e 8792795 e 8799167 norte. O objetivo do trabalho foi caracterizar a estrutura de um fragmento de floresta inundável de aproximadamente um hectare, inserido em área de orizicultura irrigada. Para o levantamento fitossociológico, foram amostrados todos os indivíduos arbustivo-arbóreos com perímetro a 1,30 m do solo (PAP > 15 cm. Ao todo, foram encontrados 807 indivíduos, 35 famílias e 70 espécies. As espécies com maior VC, em ordem decrescente, foram Hirtella racemosa Lam., Qualea multiflora Mart. e Cecropia pachystachya Trécul. As famílias mais ricas em espécies foram Fabaceae (9, Vochysiaceae (6, Annonaceae e Malvaceae (4. O índice de diversidade de Shannon (H' foi de 3,44. A distribuição de classes de diâmetro apresentou curva na forma de "J" invertido, estando a maioria dos indivíduos na primeira classe.The natural fragments of floodplain forests known as "ipucas" are located in the Araguaia plain, in form of natural depressions, which are located in areas of floodplains in the State of Tocantins, favoring its flooding and, consequently, the largest period of water retention during periods of high precipitation. The present study was developed at Lagoa Verde farm, municipal district of Lagoa da Confusão, Tocantins between the coordinates UTM: 643586 and 644060 east and 8792795 and 8799167 north. The objective of the work was to characterize the structure of a floodplain forest fragment of approximately one-hectare area

The effect of ammonium chloride concentrationson the content of carotenoid pigments in the body of Lymnaea stagnalis

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G. E. Kyrychuk

2015-09-01

Full Text Available The paper deals with the peculiarities of distribution of β-carotene and xanthophylls in the body of Lymnaea stagnalis (Linnaeus, 1758 (Pulmonata, Lymnaeidae in the normal conditions and under the action of different concentrations of NH4Cl, depending on duration of exposure (2, 7, 14 and 21 days. L. stagnalis (300 copies were collected in September 2013–2014 (in Zhytomyr region. Toxicant NH4Cl was used in concentrations corresponding to 0.5, 2, 5 and 10 MAC. The content of β-carotene and xanthophylls was determined by the Taylor method. 2400 biochemical analyses were carried out. The article shows that under the action of 0.5 MPC NH4Cl adaptive processes are stimulated in the body of L. stagnalis and total antioxidant activity is activated, as evidenced by changes in the content of β-carotene and xanthophylls in the organs (hepatopancreas, pallium and foot and tissues (hemolymph of the mollusks under study. The research proves that the action of concentration discussed above during 2, 14 and 21 days causes the increase in the content of β-carotene in hepatopancreas (1.63–5.73 times, pallium (1.68–5.37 times and foot (1.81–3.89 times of animals. Incubation of mollusks in solutions of NH4Cl (0.5 MAC for 7 days resulted in the decrease of figures in these organs by 23,6–38,8% relatively to control. It should be noted that from the overall dynamics hemolymph is somewhat distracting; for it 1.35–1.46 times’ decrease in the content of β-carotene after short-term action of the toxicant (2 and 7 days and β-carotene increase by 2.79–7.38 times under conditions of prolonged exposure (14 and 21 days is recorded. The content of xanthophylls correlates with the content of β-carotene on the 2nd, 7th and 14th day of toxic action, but by increasing the time of contact with toxic substances to 21 days the decrease in figures by 20,9–40,7% in foot and pallium is recorded. When the toxicant concentration is increased to 2 MAC (with 2

Characterization of putative virulence factors of Serratia marcescens strain SEN for pathogenesis in Spodoptera litura.

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Aggarwal, Chetana; Paul, Sangeeta; Tripathi, Vishwas; Paul, Bishwajeet; Khan, Md Aslam

2017-02-01

Two Serratia marcescens strains, SEN and ICC-4, isolated from diseased insect cadavers were observed to differ considerably in their virulence towards Spodoptera litura. The present study was aimed to characterize the possible virulence factors present in the virulent Serratia marcescens strain SEN. Both the S. marcescens strains were evaluated for the presence of various lytic enzymes such as chitinase, lipase, protease and phospholipase. The virulent S. marcescens strain SEN was observed to possess considerably higher activity of chitinase and protease enzymes; activity of phospholipase enzyme was also higher. Although, all the three toxin genes shlA, phlA and swr could be detected in both the S. marcescens strains, there was a higher expression of these genes in the virulent strain SEN. S. marcescens strain ICC-4 showed greater reduction in overall growth yield in the post-exponential phase in the presence of midgut juice and hemolymph of S. litura larvae, as compared to S. marcescens strain SEN. Proliferation of the S. marcescens strain SEN was also considerably higher in foregut, midgut and hemolymph of S. litura larvae, as compared to strain ICC-4. Peritrophic membrane treated with broth culture of the S. marcescens strain SEN showed higher damage as compared to strain ICC-4. The peritrophic membrane of larvae fed on diet treated with the virulent strain showed considerable damage while the peritrophic membrane of larvae fed on diet treated with the non-virulent strain showed no damage. This is the first report documenting the fate of ingested S. marcescens in S. litura gut and the relative expression of toxin genes from two S. marcescens strains differing in their virulence towards S. litura. Copyright © 2016 Elsevier Inc. All rights reserved.

GP-9s are ubiquitous proteins unlikely involved in olfactory mediation of social organization in the red imported fire ant, Solenopsis invicta.

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Walter S Leal

Full Text Available The red imported fire ant (RIFA, Solenopsis invicta, is an invasive species, accidentally introduced in the United States that can cause painful (sometimes life-threatening stings to human, pets, and livestock. Their colonies have two social forms: monogyne and polygyne that have a single and multiple functional queens, respectively. A major gene (Gp-9, identified as a putative pheromone-binding protein on the basis of a modest amino acid sequence identity, has been suggested to influence the expression of colony social organization. Monogyne queens are reported to possess only the GP-9B alleles, whereas polygyne queens possess both GP-9B and GP-9b. Thus, both social forms are reported to express GP-9B, with GP-9b being a marker expressed in polygynes but it is absent in monogynes. Here, we report two types of polygyne colonies, one that does not express GP-9b (monogyne-like and the other expressing both proteins, GP-9B and GP-9b. Given their expression pattern, GP-9s are hemolymph proteins, which are more likely to be involved in the transport of lipids and small ligands within the homocoel. GP-9B existed in two forms, one of them is phosphorylated. The helical-rich content of the protein resembles the secondary structures of a beetle hemolymph protein and moth pheromone-binding proteins. An olfactory role is unlikely given the lack of specific expression in the sensillar lymph. In marked contrast to GP-9s, a chemosensory protein, SinvCSP, is demonstrated to be specifically expressed in the antennae. Within the antennae, expression of SinvCSP is restricted to the last two segments, which are known to house olfactory sensilla.

[Functional morphology of blowfly Calliphora vicina hemocytes].

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Kind, T V

2012-01-01

In the hemolymph of Calliphora seven types of hemocytes were revealed. These are prohemocytes, which are the stem cells, stable and unstable hyaline cells, thrombocytoids, spindle cells, juvenile plasmatocytes and plasmatocytes I-IV, which represent sequential stages of one cell line differentiation were registered. The margin between them is completion of the crop emptying and beginning of wandering stage. In the feeding and crop emptying larvae take place rising of hyaline cells, thrombocytoids and hyaline cells amount with parallel growth of their defense function. The second wave of hemogenesis occur in the end of crop emptying period. It is accompanied by burst of plasmatocyte I production with their subsequent differentiation to plasmatocytes II-IV. Production of stable hyaline cells and respectively prothrombocytoids may be regulated not only by hormonal background but also by inorganic or organic particles invaded into the hemocel. Three types of hemocytes are involved in loosing of hemolymph from alien particles, notably thrombocytoids, juvenile plasmatocytes and plasmatocytes I and II. Thrombocytoids are responsible for parasitic eggs encapsulation. In addition they can phagocytize tiny organic and inorganic particles. Juvenile plasmatocytes respond to alien invasion almost as quickly as thrombocytoids at the onset of invasion. Plasmatocytes I and II start phagocytosis more slowly, hours post invasion, frequently accumulating the particles previously catched by thrombocytoids. Plasmatocytes I can absorb foreign particles and group in morules and can also surround filled thrombocytoids forming distinctive capsules. Both morules and capsules are temporary structures and disintegrate some hours lately. It is supposed the existence of three levels of immune defence: the fast response reaction of thrombocytoids and juvenile plasmatocytes and slow cellular reactions of plasmatocytes I. They are prerequisites for more extensive humoral response.

A novel endogenous inhibitor of phenoloxidase from Musca domestica has a cystine motif commonly found in snail and spider toxins.

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Daquinag, A C; Sato, T; Koda, H; Takao, T; Fukuda, M; Shimonishi, Y; Tsukamoto, T

1999-02-16

Phenoloxidase inhibitor (POI), found in the hemolymph of housefly pupae, is a novel dopa-containing and cystine-rich peptide that competitively inhibits phenoloxidase with a Ki in the nanomolar range. [Tyr32]POI is a potential precursor molecule also found in the hemolymph that may be posttranslationally oxidized to the dopa-containing peptide after creation of a rigid structure. By employing both a solid-phase peptide synthesis system based on a 9-fluorenylmethoxycarbonyl strategy and a specific air oxidation technique to ensure correct folding, we have been able to synthesize [Tyr32]POI. The synthetic [Tyr32]POI was confirmed to be identical to the native [Tyr32]POI by coelution high-performance liquid chromatography analysis and by enzymatic analysis using the phenoloxidase inhibition assay. To determine the disulfide pairings within the peptides, a series of enzyme hydrolyses and partial reduction/alkylation steps were performed. Three cystine pairs (Cys11-Cys25, Cys18-Cys29, and Cys24-Cys36) were determined by identification of the resulting peptides. The disulfide pairings of the two adjacent Cys residues (Cys11-Cys25 and Cys24-Cys36) were unambiguously assigned by comparing the derived fragments with the two possible isomers synthesized through a novel disulfide-linking technique. The arrangement of the disulfide bridges in POI was found to be topologically identical to those found for several peptides within the inhibitor cystine knot structural family. Although these peptides share a low primary sequence homology and display a diversity of biological functions, they nonetheless share similarities in their cystine motifs and tertiary structure. The tertiary structure model of POI, which was derived through molecular dynamics and energy minimization studies using restraints with determined disulfide connectivities, suggests that POI is a new class member of the inhibitor cystine-knot structural family.

JH III production, titers and degradation in relation to reproduction in male and female Anthonomus grandis.

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Taub-Montemayor, Tina E; Min, Kyung-Jin; Chen, Zhaorigetu; Bartlett, Terri; Rankin, Mary Ann

2005-04-01

Juvenile hormone (JH) is necessary for the production of vitellogenin (Vg) in the boll weevil, Anthonomus grandis. Occurrence of Vg in this species is typically restricted to reproductively competent females, and is not detected in untreated males. However, the JH analog, methoprene stimulates Vg production in intact males and in the isolated abdomens of both male and female boll weevils (where in each case no Vg is detected without treatment), suggesting that males are competent to produce Vg but are normally not stimulated to do so. Preliminary work indicating that male boll weevil corpora allata (CA) produced little or no JH in vitro suggested that failure of males to produce Vg might be due to very low JH levels compared to females. This study re-examines the question of JH in male boll weevils by determining in vitro production of JH III by male CA during the first 10 days after adult emergence, determining hemolymph JH esterase activity during this same time period and hemolymph JH III titers in adults of both sexes. We also re-examine the ability of isolated male abdomens to produce Vg in response to hormonal stimulation, analyzing the effect of a wide range of methoprene and JH III dosages. Results indicate that male A. grandis have circulating JH titers and JH production similar to females. JH esterase activity is slightly but significantly higher in males than females. Vg production by isolated abdomens of both sexes after stimulation with methoprene or JH III was confirmed. Dose response studies indicated that high levels of methoprene were less effective than intermediate doses in stimulating Vg synthesis in both sexes. We conclude that the sexually dimorphic effect of JH on Vg synthesis is not due to differences in JH production or differences in JH titer between the sexes.

THE EVOLUTION OF SOME BIOCHEMICAL PARAMETERS IN THE HONEYBEES’ HAEMOLYMPH (A. M. CARPATHICA COLLECTED IN THE INACTIVE SEASON

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AGRIPINA SAPCALIU

2008-10-01

Full Text Available The biochemical analyses of the blood are largely used for the routine diagnosis and especially for the metabolic survey in farm animals. These facts conduct us to the idea that similar analyses, applied on honeybee hemolymph, could be used IN monitoring the healthy state of honeybee colonies. The present studies represent preliminary researches, which aimed to investigate the variability of the main biochemical parameters in the hemolymph of the healthy honeybees (Apis mellifera in inactive season. The researches were carried out on honeybee samples collected from 5 honeybee colonies belonging to a breeding apiary of the Institute for Beekeeping Research and Development from Bucharest. In order to perform the biochemical analyses, the honeybees samples, consisting in 50 individuals on sample (10 individuals/colony were randomly collected and their haemolimph collected, at different time intervals, in inactive season (fall-winter. Totally, there were collected 250 haemolyph samples in a 2 years interval and the following 21 biochemical parameters were analysed: GLU, HDL-c, ALP, T-cho, Tprot, Alb., BUN, LDH, CPK,, Mg, IP, GGT, GOT, GPT, Ca, Cre,, Amy, T–BIL, TG, UA.. The test was carried out after the collection and processing of the samples using the SPOTCHEM EZSP4430, equipment with dry kits, the slides technique, respectively .During the 2nd part of the inactive season, the values of most biochemical parameters increase in different proportions, their levels being maintained also in the first part of the active seasons (April, May, June.The values obtained for the main studied biochemical parameters in the haemolymph of the healthy honeybees collected from honeybee colonies kept in natural conditions show a highly variable evolution in the two consecutive years of experiments during the inactive season.

CRF-like diuretic hormone negatively affects both feeding and reproduction in the desert locust, Schistocerca gregaria.

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Pieter Van Wielendaele

Full Text Available Diuretic hormones (DH related to the vertebrate Corticotropin Releasing Factor (CRF have been identified in diverse insect species. In the migratory locust, Locusta migratoria, the CRF-like DH (CRF/DH is localized in the same neurosecretory cells as the Ovary Maturating Parsin (OMP, a neurohormone that stimulates oocyte growth, vitellogenesis and hemolymph ecdysteroid levels in adult female locusts. In this study, we investigated whether CRF-like DH can influence feeding and reproduction in the desert locust, Schistocerca gregaria. We identified two highly similar S. gregaria CRF-like DH precursor cDNAs, each of which also encodes an OMP isoform. Alignment with other insect CRF-like DH precursors shows relatively high conservation of the CRF/DH sequence while the precursor region corresponding to OMP is not well conserved. Quantitative real-time RT-PCR revealed that the precursor transcripts mainly occur in the central nervous system and their highest expression level was observed in the brain. Injection of locust CRF/DH caused a significantly reduced food intake, while RNAi knockdown stimulated food intake. Therefore, our data indicate that CRF-like DH induces satiety. Furthermore, injection of CRF/DH in adult females retarded oocyte growth and caused lower ecdysteroid titers in hemolymph and ovaries, while RNAi knockdown resulted in opposite effects. The observed effects of CRF/DH may be part of a wider repertoire of neurohormonal activities, constituting an integrating control system that affects food intake and excretion, as well as anabolic processes like oocyte growth and ecdysteroidogenesis, following a meal. Our discussion about the functional relationship between CRF/DH and OMP led to the hypothesis that OMP may possibly act as a monitoring peptide that can elicit negative feedback effects.

No genetic tradeoffs between hygienic behaviour and individual innate immunity in the honey bee, Apis mellifera.

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Brock A Harpur

Full Text Available Many animals have individual and social mechanisms for combating pathogens. Animals may exhibit short-term physiological tradeoffs between social and individual immunity because the latter is often energetically costly. Genetic tradeoffs between these two traits can also occur if mutations that enhance social immunity diminish individual immunity, or vice versa. Physiological tradeoffs between individual and social immunity have been previously documented in insects, but there has been no study of genetic tradeoffs involving these traits. There is strong evidence that some genes influence both innate immunity and behaviour in social insects--a prerequisite for genetic tradeoffs. Quantifying genetic tradeoffs is critical for understanding the evolution of immunity in social insects and for devising effective strategies for breeding disease-resistant pollinator populations. We conducted two experiments to test the hypothesis of a genetic tradeoff between social and individual immunity in the honey bee, Apis mellifera. First, we estimated the relative contribution of genetics to individual variation in innate immunity of honey bee workers, as only heritable traits can experience genetic tradeoffs. Second, we examined if worker bees with hygienic sisters have reduced individual innate immune response. We genotyped several hundred workers from two colonies and found that patriline genotype does not significantly influence the antimicrobial activity of a worker's hemolymph. Further, we did not find a negative correlation between hygienic behaviour and the average antimicrobial activity of a worker's hemolymph across 30 honey bee colonies. Taken together, our work indicates no genetic tradeoffs between hygienic behaviour and innate immunity in honey bees. Our work suggests that using artificial selection to increase hygienic behaviour of honey bee colonies is not expected to concurrently compromise individual innate immunity of worker bees.

Identification and Characterization of 30 K Protein Genes Found in Bombyx mori (Lepidoptera: Bombycidae) Transcriptome

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Shi, Xiao-Feng; Li, Yi-Nü; Yi, Yong-Zhu; Xiao, Xing-Guo; Zhang, Zhi-Fang

2015-01-01

The 30 K proteins, the major group of hemolymph proteins in the silkworm, Bombyx mori (Lepidoptera: Bombycidae), are structurally related with molecular masses of ∼30 kDa and are involved in various physiological processes, e.g., energy storage, embryonic development, and immune responses. For this report, known 30 K protein gene sequences were used as Blastn queries against sequences in the B. mori transcriptome (SilkTransDB). Twenty-nine cDNAs (Bm30K-1–29) were retrieved, including four being previously unidentified in the Lipoprotein_11 family. The genomic structures of the 29 genes were analyzed and they were mapped to their corresponding chromosomes. Furthermore, phylogenetic analysis revealed that the 29 genes encode three types of 30 K proteins. The members increased in each type is mainly a result of gene duplication with the appearance of each type preceding the differentiation of each species included in the tree. Real-Time Quantitative Polymerase Chain Reaction (Q-PCR) confirmed that the genes could be expressed, and that the three types have different temporal expression patterns. Proteins from the hemolymph was separated by SDS-PAGE, and those with molecular mass of ∼30 kDa were isolated and identified by mass spectrometry sequencing in combination with searches of various databases containing B. mori 30K protein sequences. Of the 34 proteins identified, 13 are members of the 30 K protein family, with one that had not been found in the SilkTransDB, although it had been found in the B. mori genome. Taken together, our results indicate that the 30 K protein family contains many members with various functions. Other methods will be required to find more members of the family. PMID:26078299

The oxidative stress and antioxidant responses of Litopenaeus vannamei to low temperature and air exposure.

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Xu, Zihan; Regenstein, Joe M; Xie, Dandan; Lu, Wenjing; Ren, Xingchen; Yuan, Jiajia; Mao, Linchun

2018-01-01

Low temperature and air exposure were the key attributes for waterless transportation of fish and shrimp. In order to investigate the oxidative stress and antioxidant responses of the live shrimp Litopenaeus vannamei in the mimic waterless transportation, live shrimp were cooled at 13 °C for 3 min, stored in oxygen at 15 °C for 12 h, and then revived in water at 25 °C. The survival rate of shrimp under this waterless transportation system was over 86.67%. The ultrastructure of hepatopancreas cells were observed while activities of superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), glutathione peroxidase (GSH-Px), antisuperoxide anion free radicals (ASAFR), total antioxidant capacity (TAOC), reactive oxygen species (ROS) production, content of malondialdehyde (MDA) and relative mRNA expressions of CAT and GSH-Px in the hemolymph and hepatopancreas were determined. Slight distortions of some organelles in hepatopancreas cells was reversible upon the shrimp revived from the cold shock. The activities of SOD, POD, CAT, GSH-Px, TAOC, ROS production and relative mRNA expressions of CAT and GSH-Px increased following the cold shock and reached peak levels after 3 or 6 h of storage, and then decreased gradually. There was no significant difference between the fresh and the revived shrimp in SOD, POD, GSH-Px, TAOC, ROS, MDA and relative mRNA expressions of CAT and GSH-Px. The oxidative stress and antioxidant responses were tissue-specific because hepatopancreas seemed to have a greater ability to defend against organelle damage and was more sensitive to stress than hemolymph based on the results of SOD activity, MDA content and GSH-Px mRNA expression. These results revealed that low temperature and air exposure caused significant oxidative and antioxidant responses, but did not lead to irreversible damages in this waterless system. Copyright © 2017 Elsevier Ltd. All rights reserved.

Protein levels and colony development of Africanized and European honey bees fed natural and artificial diets.

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Morais, M M; Turcatto, A P; Pereira, R A; Francoy, T M; Guidugli-Lazzarini, K R; Gonçalves, L S; de Almeida, J M V; Ellis, J D; De Jong, D

2013-12-19

Pollen substitute diets are a valuable resource for maintaining strong and health honey bee colonies. Specific diets may be useful in one region or country and inadequate or economically unviable in others. We compared two artificial protein diets that had been formulated from locally-available ingredients in Brazil with bee bread and a non-protein sucrose diet. Groups of 100 newly-emerged, adult workers of Africanized honey bees in Brazil and European honey bees in the USA were confined in small cages and fed on one of four diets for seven days. The artificial diets included a high protein diet made of soy milk powder and albumin, and a lower protein level diet consisting of soy milk powder, brewer's yeast and rice bran. The initial protein levels in newly emerged bees were approximately 18-21 µg/µL hemolymph. After feeding on the diets for seven days, the protein levels in the hemolymph were similar among the protein diet groups (~37-49 µg/µL after seven days), although Africanized bees acquired higher protein levels, increasing 145 and 100% on diets D1 and D2, respectively, versus 83 and 60% in the European bees. All the protein diets resulted in significantly higher levels of protein than sucrose solution alone. In the field, the two pollen substitute diets were tested during periods of low pollen availability in the field in two regions of Brazil. Food consumption, population development, colony weight, and honey production were evaluated to determine the impact of the diets on colony strength parameters. The colonies fed artificial diets had a significant improvement in all parameters, while control colonies dwindled during the dearth period. We conclude that these two artificial protein diets have good potential as pollen substitutes during dearth periods and that Africanized bees more efficiently utilize artificial protein diets than do European honey bees.

INSECTICIDAL AND OXIDATIVE EFFECTS OF AZADIRACHTIN ON THE MODEL ORGANISM Galleria mellonella L. (LEPIDOPTERA: PYRALIDAE).

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Dere, Beyza; Altuntaş, Hülya; Nurullahoğlu, Z Ulya

2015-07-01

The insecticidal effects, specifically, changes in hemolymph total protein and malondialdehyde (MDA) levels, and antioxidant enzyme activities of azadirachtin (AZA) given to the wax moth, Galleria mellonella L. (Lepidoptera: Pyralidae) larvae via force feeding were investigated. Bioassays showed that the LD50 and LD99 (lethal dose) values of AZA were 2.1 and 4.6 μg/larva, respectively. Experimental analyses were performed with five doses of AZA (0.5, 1, 1.5, 2, and 3 μg/larva). Total protein level in larval hemolymph increased at all AZA doses at 24 h whereas a considerable decrease was observed at 2 and 3 μg/larva doses, and only an increase displayed at 1.5 μg/larva at 72 h. The level of MDA increased at 2 and 3 μg/larva doses at 24 h compared with controls. This trend was also observed at 1.5, 2, and 3 μg/larva doses at 72 h and MDA levels were lower when compared with those of 24 h at all doses except for 1.5 μg/larva dose. Catalase activity decreased at 1, 1.5, and 2 μg/larva doses at 24 h whereas increased at all doses except for 0.5 μg/larva at 72 h compared with controls. AZA led to a decline in superoxide dismutase activity at all experimental doses at 24 and 72 h except for 3 μg/larva doses at 72 h. An increase in glutathione-S-transferase (GST) activity was evident at all AZA doses at 24 h. AZA displayed 68% decline in GST activity at 72 h post treatments when compared to 24 h. Consequently, We infer that the toxicity of AZA extends beyond its known actions in molting processes to redox homeostasis. © 2015 Wiley Periodicals, Inc.

Evaluation of uptake and chronic toxicity of virgin polystyrene microbeads in freshwater zebra mussel Dreissena polymorpha (Mollusca: Bivalvia).

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Magni, Stefano; Gagné, François; André, Chantale; Della Torre, Camilla; Auclair, Joëlle; Hanana, Houda; Parenti, Camilla Carla; Bonasoro, Francesco; Binelli, Andrea

2018-08-01

Microplastics (MPs), plastic debris smaller than 5mm, are widely found in both marine and freshwater ecosystems. However, few studies regarding their hazardous effects on inland water organisms, have been conducted. For this reason, the aim of our research was the evaluation of uptake and chronic toxicity of two mixtures (MIXs) of virgin polystyrene microbeads (PMs) of 10μm and 1μm in size (MIX 1, with 5×10 5 of 1μmsizePMs/L and 5×10 5 of 10μmsizePMs/L, and MIX 2 with 2×10 6 of 1μmsizePMs/L and 2×10 6 of 10μmsizePMs/L) on freshwater zebra mussel Dreissena polymorpha (Mollusca: Bivalvia) during 6 exposure days. The PM uptake in the mussel body and hemolymph was assessed using confocal microscopy, while the chronic toxicity of PMs was evaluated on exposed mussels using a comprehensive battery of biomarkers of cellular stress, oxidative damage and neuro- genotoxicity. Confocal microscopy analyses showed that MPs concentrated in the gut lumen of exposed mussels, absorbed and transferred firstly in the tissues and then in the hemolymph. The results revealed that PMs do not produce oxidative stress and genetic damage, with the exception of a significant modulation of catalase and glutathione peroxidase activities in mussels exposed to MIX 1. Regarding neurotoxicity, we observed only a significant increase of dopamine concentration in mussels exposed to both MIXs, suggesting a possible implication of this neurotransmitter in an elimination process of accumulated PMs. This research represents a first study about the evaluation of virgin MP toxicity in zebra mussel and more research is warranted concerning the long term neurological effects of virgin MPs. Crown Copyright © 2018. Published by Elsevier B.V. All rights reserved.

Isotopic incorporation rates and discrimination factors in mantis shrimp crustaceans.

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Maya S deVries

Full Text Available Stable isotope analysis has provided insights into the trophic ecology of a wide diversity of animals. Knowledge about isotopic incorporation rates and isotopic discrimination between the consumer and its diet for different tissue types is essential for interpreting stable isotope data, but these parameters remain understudied in many animal taxa and particularly in aquatic invertebrates. We performed a 292-day diet shift experiment on 92 individuals of the predatory mantis shrimp, Neogonodactylus bredini, to quantify carbon and nitrogen incorporation rates and isotope discrimination factors in muscle and hemolymph tissues. Average isotopic discrimination factors between mantis shrimp muscle and the new diet were 3.0 ± 0.6 ‰ and 0.9 ± 0.3 ‰ for carbon and nitrogen, respectively, which is contrary to what is seen in many other animals (e.g. C and N discrimination is generally 0-1 ‰ and 3-4 ‰, respectively. Surprisingly, the average residence time of nitrogen in hemolymph (28.9 ± 8.3 days was over 8 times longer than that of carbon (3.4 ± 1.4 days. In muscle, the average residence times of carbon and nitrogen were of the same magnitude (89.3 ± 44.4 and 72.8 ± 18.8 days, respectively. We compared the mantis shrimps' incorporation rates, along with rates from four other invertebrate taxa from the literature, to those predicted by an allometric equation relating carbon incorporation rate to body mass that was developed for teleost fishes and sharks. The rate of carbon incorporation into muscle was consistent with rates predicted by this equation. Our findings provide new insight into isotopic discrimination factors and incorporation rates in invertebrates with the former showing a different trend than what is commonly observed in other animals.

A strategy for solubilizing delipidated apolipoprotein with lysophosphatidylcholine and reconstitution with phosphatidylcholine

International Nuclear Information System (INIS)

Kawooya, J.K.; Wells, M.A.; Law, J.H.

1989-01-01

The apolipoproteins of insect lipophorin were dissociated in guanidinium chloride and isolated by gel permeation chromatography. Over 98% of the total lipid in lipophorin was associated with apolipophorin I (apoLp-I), thus suggesting this apolipoprotein to be the lipid binding component of the particle. ApoLp-I was delipidated with ethanol/ether and solubilized in buffer that contained radioactive lysophosphatidylcholine ([ 3 H]LPC) above the critical micellar concentration. Sonic irradiation of radioactive phosphatidylcholine ([ 14 C]PC) with [ 3 H]LPC-solubilized apoLp-I at a molar ratio of 318 resulted in reconstituted lipophorin I (RLp-I). [ 3 H]LPC was bound to fatty acid free bovine serum albumin and was separated from RLp-I by density gradient ultracentrifugation and gel permeation chromatography. Negatively stained RLp-I particles were quasispherical with an average radius of 55 angstrom, and their overall morphology and secondary structure were similar to those of native hemolymph lipophorin. The RLp-I particle had a ρ = 1.137 g/mL, a M r ∼ 5.2 x 10 5 , and a [ 14 C]PC:apoLp-I molar ratio of 308. From the compositional analysis, molecular size, trypsinization, and lipolysis with phospholipase A 2 , the authors concluded that each RLp-I particle contained one molecule of apoLp-I and a monomolecular layer of [ 14 C]PC. When injected into the hemolymph of adult moths in vivo, RLp-I was loaded with lipid, as judged by a decrease in its density both in the presence and in the absence of adipokinetic hormone. The similarities in morphology and immunology of RLp-I and native lipophorin, together with the ability of RLp-I to load lipid, suggest that reconstituted lipophorins may serve as models to probe lipophorin structure and function

Cuticular bacteria appear detrimental to social spiders in mixed but not monoculture exposure

Science.gov (United States)

Keiser, Carl N.; Shearer, Taylor A.; DeMarco, Alexander E.; Brittingham, Hayley A.; Knutson, Karen A.; Kuo, Candice; Zhao, Katherine; Pruitt, Jonathan N.

2016-01-01

Abstract Much of an animal’s health status, life history, and behavior are dictated by interactions with its endogenous and exogenous bacterial communities. Unfortunately, interactions between hosts and members of their resident bacterial community are often ignored in animal behavior and behavioral ecology. Here, we aim to identify the nature of host–microbe interactions in a nonmodel organism, the African social spider Stegodyphus dumicola. We collected and identified bacteria from the cuticles of spiders in situ and then exposed spiders to bacterial monocultures cultures via topical application or injection. We also topically inoculated spiders with a concomitant “cocktail” of bacteria and measured the behavior of spiders daily for 24 days after inoculation. Lastly, we collected and identified bacteria from the cuticles of prey items in the capture webs of spiders, and then fed spiders domestic crickets which had been injected with these bacteria. We also injected 1 species of prey-borne bacteria into the hemolymph of spiders. Only Bacillus thuringiensis caused increased mortality when injected into the hemolymph of spiders, whereas no bacterial monocultures caused increased mortality when applied topically, relative to control solutions. However, a bacterial cocktail of cuticular bacteria caused weight loss and mortality when applied topically, yet did not detectibly alter spider behavior. Consuming prey injected with prey-borne bacteria was associated with an elongated lifespan in spiders. Thus, indirect evidence from multiple experiments suggests that the effects of these bacteria on spider survivorship appear contingent on their mode of colonization and whether they are applied in monoculture or within a mixed cocktail. We urge that follow-up studies should test these host–microbe interactions across different social contexts to determine the role that microbes play in colony performance. PMID:29491926

Preparation of a specifically tritiated locust adipokinetic hormone analog with full biological potency

Energy Technology Data Exchange (ETDEWEB)

Muramoto, K; Ramachandran, J; Moshitzky, P; Applebaum, S W [Hormone Research Laboratory and Department of Biochemistry and Biophysics, University of California, San Francisco, CA, USA and Department of Entomology, The Hebrew University of Jerusalem, Rehovot, Israel

1984-01-01

A synthetic peptide related to locus adipokinetic hormone (AKH) and shrimp red pigment concentrating hormone (RPCH) containing a tyrosine residue in place of phenylalanine was iodinated and the 3,5-diiodotyrosyl derivative was isolated by reverse phase HPLC. Catalytic dehalogenation of the diiodo derivative in the presence of tritium yielded the tritiated AKH analog which was isolated by gel filtration on Sephadex LH-20 and reverse phase HPLC. The tritiated peptide was formed to be identical to AKH in its ability to stimulate lipid release into the hemolymph of locusts in vivo where the diiodotryrosyl derivative was inactive. The specific radioactivity of the tritiated peptide was 57.2 Ci/mmol, or 99% of the theoretical value.

Cellular tropism, population dynamics, host range and taxonomic status of an aphid secondary symbiont, SMLS (Sitobion miscanthi L type symbiont.

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Tong Li

Full Text Available SMLS (Sitobion miscanthi L type symbiont is a newly reported aphid secondary symbiont. Phylogenetic evidence from molecular markers indicates that SMLS belongs to the Rickettsiaceae and has a sibling relationship with Orientia tsutsugamushi. A comparative analysis of coxA nucleotide sequences further supports recognition of SMLS as a new genus in the Rickettsiaceae. In situ hybridization reveals that SMLS is housed in both sheath cells and secondary bacteriocytes and it is also detected in aphid hemolymph. The population dynamics of SMLS differ from those of Buchnera aphidicola and titer levels of SMLS increase in older aphids. A survey of 13 other aphids reveals that SMLS only occurs in wheat-associated species.

Fast and Cost-Effective Biochemical Spectrophotometric Analysis of Solution of Insect “Blood” and Body Surface Elution

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Aleksandra Łoś

2018-05-01

Full Text Available Using insect hemolymph (“blood” and insect body surface elutions, researchers can perform rapid and cheap biochemical analyses to determine the insect’s immunology status. The authors of this publication describe a detailed methodology for a quick marking of the concentration of total proteins and evaluation of the proteolytic system activity (acid, neutral, and alkaline proteases and protease inhibitors, as well as a methodology for quick “liver” tests in insects: alanine aminotransferase (ALT, aspartate aminotransferase (AST, alkaline phosphatase (ALP, and urea and glucose concentration analyses. The meaning and examples of an interpretation of the results of the presented methodology for biochemical parameter determination are described for the example of honey bees.

[Hemoglobins, XXXII. Analysis of the primary structure of the monomeric hemoglobin CTT VIIA (erythrocruorin) or Chironomus thummi thummi, Diptera (author's transl)].

Science.gov (United States)

Kleinschmidt, T; Braunitzer, G

1980-01-01

The dimeric hemoglobin CTT VIIA (erythrocruorin) was isolated from the hemolymph of the larva from Chironomus thummi thummi and purified by preparative polyacrylamide gel electrophoresis. Peptides obtained by limited tryptical digestion were sequenced by automatic Edman degradation. For the elucidation of the sequence in the C-terminal region of the chain, additional cleavages with proteinase of Staphylococcus aureus and chymotrypsin were necessary. CTT VIIA is compared with human beta-chains and other hemoglobins of Chironomus. The amino acid residues in the pocket are especially discussed. Most of them are invariant in all Chironomus hemoglobins, independent of the size of the heme pocket, which is normal in some components and enlarged in others.

Plants used traditionally to treat malaria in Brazil: the archives of Flora Medicinal

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Botsaris Alexandros S

2007-05-01

Full Text Available Abstract The archives of Flora Medicinal, an ancient pharmaceutical laboratory that supported ethnomedical research in Brazil for more than 30 years, were searched for plants with antimalarial use. Forty plant species indicated to treat malaria were described by Dr. J. Monteiro da Silva (Flora Medicinal leader and his co-workers. Eight species, Bathysa cuspidata, Cosmos sulphureus, Cecropia hololeuca, Erisma calcaratum, Gomphrena arborescens, Musa paradisiaca, Ocotea odorifera, and Pradosia lactescens, are related as antimalarial for the first time in ethnobotanical studies. Some species, including Mikania glomerata, Melampodium divaricatum, Galipea multiflora, Aspidosperma polyneuron, and Coutarea hexandra, were reported to have activity in malaria patients under clinical observation. In the information obtained, also, there were many details about the appropriate indication of each plant. For example, some plants are indicated to increase others' potency. There are also plants that are traditionally employed for specific symptoms or conditions that often accompany malaria, such as weakness, renal failure or cerebral malaria. Many plants that have been considered to lack activity against malaria due to absence of in vitro activity against Plasmodium can have other mechanisms of action. Thus researchers should observe ethnomedical information before deciding which kind of screening should be used in the search of antimalarial drugs.

Tropical rainforest biome of Biosphere 2. Structure, composition and results of the first 2 years of operation

Energy Technology Data Exchange (ETDEWEB)

Leigh, Linda S. [Systems Ecology and Energy Analysis Program, Department of Environmental Engineering Sciences, University of Florida, Gainesville, FL (United States); Burgess, Tony; Marino, Bruno D.V.; Wei, Yong Dan [Biosphere 2 Center, Inc. P.O. Box 689, Oracle, AZ (United States)

1999-06-01

The tropical rainforest biome in the Biosphere 2 mesocosm was managed with rainfall and temperature conditions to simulate a natural rainforest typical of the new world tropics. The establishment of the biome was based on the introduction of excessive numbers of species allowing self-organization of an ecologically unique rainforest. Over 282 species of plants from rainforest areas were planted within the topographically diverse rainforest biome (area of 1900 m{sup 2}, volume of 35,000 m{sup 3}), just before the Biosphere 2 closure in 1991. Approximately 61% of these species survived when counted in 1993, representing a plant species richness reduction to 172 species in 0.19 hectare. Rank order graphs show that a high diversity community resulted not unlike insular rainforests. The plants of the rainforest mesocosm, however, grew under anomalous conditions of soil (amended desert grassland soil), atmospheric composition (CO{sub 2} up to 4500 ppm by volume (ppmv)) and rainwater composition (high salinity and nutrients). Stem growth rates of a dominant canopy tree, Cecropia, were up to four times higher but had reduced diameter at breast height compared to natural counterparts. Human intervention in plant succession was also an important factor in shaping the ecology of the rainforest biome of Biosphere 2

Non-experimental validation of ethnoveterinary plants and indigenous knowledge used for backyard pigs and chickens in Trinidad and Tobago.

Science.gov (United States)

Lans, C; Georges, K; Brown, G

2007-06-01

This paper presents the findings of an exploratory study on ethnoveterinary medicines used for backyard pigs and backyard chickens in Trinidad and Tobago. Research data was collected from 1995 to September 2000. Six plants are used for backyard pigs. Crushed leaves of immortelle (Erythrina pallida, E. micropteryx) are used to remove dead piglets from the uterus. Leaf decoctions of bois canôt (Cecropia peltata) and bamboo (Bambusa vulgaris) are used for labour pains or leaves are fed as a postpartum cleanser. Boiled green papaya fruit (Carica papaya) is fed to pigs to induce milk let-down. The leaves and flowers of male papaya plants (Carica papaya) are fed to deworm pigs. Sour orange juice (Citrus aurantium) is given to pigs to produce lean meat, and coffee grounds are used for scours. Eyebright and plantain leaves (Plantago major) are used for eye injuries of backyard chickens. Worm grass (Chenopodium ambrosioides) and cotton bush (Gossypium species) are used as anthelmintics. Aloe gel (Aloe vera) is used for internal injuries and the yellow sap from the cut Aloe vera leaf or the juice of Citrus limonia is used to purge the birds. A literature review revealed few toxicity concerns and the potential usefulness of the plants.

The potential sensitivity of tropical plants to increased ultraviolet-B radiation

International Nuclear Information System (INIS)

Ziska, L.H.

1996-01-01

Little is known concerning the impact of stratospheric ozone depletion and increasing ultraviolet (UV)-B radiation on the phenology and growth of tropical plants. This is because, ostensibly, tropical plants are already exposed to relatively high levels of UV-B radiation (relative to a temperate environment) and should, therefore, possess a greater degree of tolerance to increased UV-B radiation. In this brief review I hope to show that, potentially, direct and indirect effects on photosynthesis, assimilate partitioning, phenology and biomass could occur in both tropical crops (e.g. cassava, rice) and native species (e.g. Cecropia obtusifolia (Bertol. Fl)., Tetramolopium humile (Gray), Nana sandwicensis L.). However, it should be noted that differences in sensitivity to UV-B radiation can be related to experimental conditions, and care should be taken to ensure that the quantity and quality of background solar radiation remains at near ambient conditions. Nevertheless, by integrating current and past studies on the impact of UV-B radiation on tropical species, I hope to be able to demonstrate that photosynthesis, morphology and growth in tropical plants could be directly affected by UV-B radiation and that UV-B radiation may be a factor in species and community dynamics in natural plant populations in the tropics

Self-anointing behavior in free-ranging spider monkeys (Ateles geoffroyi) in Mexico.

Science.gov (United States)

Laska, Matthias; Bauer, Verena; Salazar, Laura Teresa Hernandez

2007-04-01

During 250 h of observation, a total of 20 episodes of self-anointing, that is, the application of scent-bearing material onto the body, were recorded in a group of free-ranging Mexican spider monkeys (Ateles geoffroyi). The animals used the leaves of three species of plants (Brongniartia alamosana, Fabaceae; Cecropia obtusifolia, Cecropiaceae; and Apium graveolens, Umbelliferae) two of which have not been reported so far in this context in any New World primate species. The findings that only two males displayed self-anointing, that only the sternal and axillary regions of the body were rubbed with the mix of saliva and plant material, and a lack of correlation between the occurrence of self-anointing and time of day, season of the year, ambient temperature or humidity do not fit the hypothesis that this behavior functions in repelling insects and/or mitigating topical skin infections in this species. Rather, the data and the observation that the leaves of all three plant species spread an intensive and aromatic odor when crushed, support the hypothesis that self-anointing in A. geoffroyi may play a role in the context of social communication, possibly for signaling of social status or to increase sexual attractiveness.

Geographical and Botanical Origin of Apis mellifera (Apidae) Honey in four Colombian Departments

International Nuclear Information System (INIS)

Nates Parra, Guiomar; Montoya, Paula Maria; Chamorro, Fermin J; Ramirez, Nedy; Giraldo, Catalina; Obregon, Diana

2013-01-01

The aim of this work was to find palynological markers which permit differentiate honeys from the departments of Boyaca, Cundinamarca, Santander and Magdalena, by its geographical and botanical origin. Melissopalynological analyses were made of 184 honey samples obtained from 131 localities. A discriminant analysis and comparisons between the species composition of honey samples were made to find geo-graphical and botanical origin differences. A total of 297 pollen species distributed in 69 families was found, being Mimosa sp., Cecropia sp., Eucalyptus sp., Piper sp. and Quercus humboldtii the most representatives. The major families were Fabaceae, Asteraceae, Myrtaceae, Rubiaceae, Fagaceae and Melastomataceae. Six honey groups differentiated by its geographical origin were found: Altiplano Cundiboyacense, Medio Chicamocha, Sumapaz, Bajo Chicamocha, Sierra Nevada de Santa Marta and Comunera Province. In a broader scale, honeys from the Andean and sub-Andean regions could be differentiated as well. Between the honey types differentiated by its botanical origin, the most important were monofloral honeys of Trifolium pratense, Coffea arabica, Eucalyptus sp., Inga sp. and Heliocarpus americanus, Asteraceae oligofloral honeys and mixtures of Q. humboldtii honeydew and floral nectar (Eucalyptussp., Brassicaceae Type, Asteraceae). This information in addition to the obtained by physico-chemical and sensorial analysis, may be the basis to acquire honeys' origin denomination.

Adipokinetic hormone-induced enhancement of antioxidant capacity of Pyrrhocoris apterus hemolymph in response to oxidative stress

Czech Academy of Sciences Publication Activity Database

Večeřa, J.; Krishnan, Natraj; Alquicer, Glenda; Kodrík, Dalibor; Socha, Radomír

2007-01-01

Roč. 146, - (2007), s. 336-342 ISSN 1532-0456 R&D Projects: GA ČR GA522/07/0788 Institutional research plan: CEZ:AV0Z50070508 Keywords : adipokinetic hormone * antioxidant activity * oxidative stress Subject RIV: ED - Physiology Impact factor: 2.345, year: 2007

Feeding ecology of the Green-cheeked parakeet (Pyrrhura molinae in dry forests in western Brazil Ecologia alimentar da Tiriba-de-cara-suja (Pyrrhura molinae em matas secas do oeste brasileiro

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J. Ragusa-Netto

2007-05-01

Full Text Available The genus Pyrrhura includes small to medium-sized parakeets, which inhabit both low and dry alongside tall rainforests, mainly in South America. Pyrrhura molinae is still common, year round, in the markedly seasonal forests of western Brazil. This parakeet, as well as most Neotropical parrots, continues to be poorly understood. Hence, in the present study I examined their foraging ecology both in a highly deciduous and in a semi-deciduous forest in western Brazil. In addition, I assessed the relationship between food resource production (flowers and fruits, and the diet of this parakeet. Pyrrhura molinae exhibited a flexible diet consisting of 16 tree species, from which it consumed flowers (three species, seeds (three species, fruit pulp or aril (four species, and both pulp and seeds (six species. Parakeets consumed a wide array of fleshy fruits in the semi-deciduous forest, especially Cecropia pachystachya catkins. Conversely, in the highly deciduous forest they extensively foraged for figs (70% of the diet, in addition to nectar and seeds from dry fruits. Ficus calyptroceras, besides being abundant, bore fruits year round, and was substantially used by parakeets every month. Potentially, by exploiting a diverse set of plant food resources, and particularly due to the substantial use of figs, asynchronously produced, Pyrrhura molinae persists during the long dry season in the markedly seasonal forests of western Brazil.O gênero Pyrrhura é constituido de pequenos periquitos comuns tanto em matas secas quanto úmidas, sobretudo da América do Sul. Pyrrhura molinae ocorre durante o ano todo em florestas altamente sazonais do oeste brasileiro. Essa espécie, bem como a maioria dos psitacídeo, permanece pouco conhecida. Portanto, nesse estudo, foi examinada a ecologia alimentar de P. molinae em dois tipos de florestas secas (altamente decídua e semidecídua, do oeste brasileiro, bem como as relações entre a produção de flores frutos e a

Site of water vapor absorption in the desert cockroach, Arenivaga investigata.

Science.gov (United States)

O'Donnell, M J

1977-01-01

The desert cockroach, Arenivaga investigata, can gain weight by absorption of water-vapor from unsaturated atmospheres above 82.5% relative humidity. Blocking the anus or the dorsal surface with wax does not prevent water vapor uptake, but interference with movements of the mouthparts or blocking the mouth with wax-prevents such uptake. Weight gains are associated with the protrusion from the mouth of two bladder-like extensions of the hypopharynx. During absorption these structures are warmer than the surrounding mouthparts, their surface temperature increasing with relative humidity. This suggests that the surfaces of the bladder-like structures function at least as sites for condensation of water vapor, but the precise location of its transfer into the hemolymph has not yet been identified. Images PMID:266217

Pesquisa de Rickettsia spp em carrapatos Amblyomma cajennense e Amblyomma dubitatum no Estado de São Paulo Survey of Rickettsia spp in the ticks Amblyomma cajennense and Amblyomma dubitatum in the State of São Paulo

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Richard Campos Pacheco

2009-06-01

Full Text Available Foi pesquisada a presença de riquétsias em 3.545 carrapatos Amblyomma cajennense e 2.666 Amblyomma dubitatum. Através do teste de hemolinfa, reação em cadeia pela polimerase e isolamento de rickettsia em cultivo celular, todos os Amblyomma cajennense foram negativos, sendo que 634 (23,8% Amblyomma dubitatum mostraram-se infectados com Rickettsia bellii.The presence of rickettsial infection was surveyed in 3,545 Amblyomma cajennense ticks and 2,666 Amblyomma dubitatum ticks. Using the hemolymph test, polymerase chain reaction and isolation of Rickettsia in cell cultures, all of the Amblyomma cajennense were negative, whereas 634 (23.8% of the Amblyomma dubitatum ticks were shown to be infected with Rickettsia bellii.

Cellular Immune Reactions of the Sunn Pest, Eurygaster integriceps, to the Entomopathogenic Fungus, Beauveria bassiana and Its Secondary Metabolites

Science.gov (United States)

Zibaee, Arash; Bandani, Ali Reza; Talaei-Hassanlouei, Reza; Malagoli, Davide

2011-01-01

In this study, five morphological types of circulating hemocytes were recognized in the hemolymph of the adult sunn pest, Eurygaster integriceps Puton (Hemiptera: Scutelleridae), namely prohemocytes, plasmatocytes, granulocytes, adipohemocytes, and oenocytoids. The effects of the secondary metabolites of the entomopathogenic fungus Beauveria bassiana on cellular immune defenses of Eurygaster integriceps were investigated. The results showed that the fungal secondary metabolites inhibited phagocytic activity of E. integriceps hemocytes and hampered nodule formation. A reduction of phenoloxidase activity was also observed. The data suggest that B. bassiana produce secondary metabolites that disable several immune mechanisms allowing the fungus to overcome and then kill its host. This characteristic makes B. bassiana a promising model for biological control of insect pests such as E. integriceps. PMID:22233481

Oxygen and carbon dioxide transporting qualities of hemocyanin in the hemolymph of a natant decapod Palaemon adspersus

DEFF Research Database (Denmark)

Weber, Roy E.; Hagerman, L.

1981-01-01

, pH dependent Bohr factor (=logP 50/pH=-2.0 and -0.9 at pH 7.85 and 7.4, respectively) (Figs. 1 and 2). These qualities are distinct from those typifying reptant hemocyanins and appear illsuited for O2 transport at low ambient tensions, but well-adapted for O2 delivery in tissues at high PO2...

Citotoxicidad de extractos de plantas medicinales sobre la línea celular de carcinoma de pulmón humano A549

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Alexis Díaz García

2011-03-01

Full Text Available OBJETIVO: evaluar el efecto de 10 extractos de plantas medicinales sobre el crecimiento de la línea celular humana de carcinoma de pulmón A549. METODOS: el efecto de los extractos sobre la células tumorales se midió a través de un ensayo colorimétrico mediante el empleo del bromuro de 3-(4,5-dimetil-tiazol-2-yl-2,5-difenil tetrazolio a concentraciones entre 3,9-250 µg/mL durante 72 h y se calculó la concentración citotóxica media para cada uno. RESULTADOS: del total de los extractos evaluados solo cuatro (Parthenium hysterophorus, Bixa orellana, Momordica charantia y Cucurbita maxima evidenciaron concentraciones citotóxicas medias inferiores a 100 µg/mL. Excepto Parthenium hysterophorus, las restantes se emplean en la medicina tradicional para el tratamiento del cáncer. Los extractos de Cecropia peltata, Melia azedarach, Annona glabra, Artemisia absintium, Lepidium virginicum y Bidens pilosa no mostraron efectos citotóxicos significativos. CONCLUSIONES: Los extractos de plantas que se emplean en la medicina tradicional para el tratamiento del cáncer, mostraron citotoxicidad sobre las células tumorales. El conocimiento etnobotánico representa una herramienta importante en la selección de plantas medicinales, en la búsqueda de nuevos compuestos para el tratamiento del cáncer.

Dieta de Traupíneos (Passeriformes, Emberizidae no Parque Estadual do Ibitipoca, Minas Gerais, Brasil Diet of Tanagers (Passeriformes, Emberizidae in Ibitipoca State Park, Minas Gerais, Brazil

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Marco A. Manhães

Full Text Available Eleven species of tanagers in Ibitipoca State Park, Minas Gerais, southeastern Brazil, were studied for 12 months and the birds diet was described. Fruits of 52 plant species were the main item consumed for all tanagers, representing 59.70% of all items in 1330 events, although different proportions in the use of this resource have been found among the bird species. The main plant families found were Myrtaceae with eight species accounting for 22.29% (n=177, and Melastomataceae with seven species accounting for 22.29% (n=177 of fruit intake. The most frequent plant species in the diet was Cecropia glaziovi Snethl. (Cecropiaceae, which made up 17.76% (n=141 of fruit intake of all species of tanagers. Arthropod consumption came second with 22.63% (n=301, but flowers, leaves, nectar, food remains left by tourists and galls were also represented. Several environmental features influenced diet composition. An Asteraceae, Vanillosmopsis erythropappa Schultz, very common in the park, was important for some birds as its flowers attracted arthropods and its branches and leaves were used as a substrate for insectivory. Fruit intake rate, some bird-plant interaction and features of food-handling were also discussed, pointing to the importance of these birds for the structure of the heterogeneous local landscape, possibly through the spread of seeds of different plant species.

Catálogo sistemático dos polens das plantas arbóreas do Brasil meridional: XXIII. Moraceae Catalogue of southern Brazilian pollen grains: XXIII. Moraceae

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Ortrud Monika Barth

1976-01-01

Full Text Available Foi estudada a morfologia polínica de espécies pertencentes a oito gêneros. Os diferentes tipos polínicos econtrados, correspondendo aos gêneros, bem como as variações das estruturas dentro de alguns deles, confirmam as relações filogenéticas nesta família. Os de morfologia mais simples correspondem a árvores ou arbustos dos gêneros Brosimum, Cecropia, Chlorophora e Coussapoa, seguindo-se Ficus, Pourouma e Sorocea; os grãos do gênero herbáceo Dorstenia apresentam uma complexa morfologia polínica. A estrutura dos esporodermas e das aberturas dos grãos de pólen comprova o seu relacionamento com outras famílias das Urticales.The pollen morphology from some species of eight genera of Moraceae has been studied. The different pollen types found, corresponding to the genera, as well as the structural variations in some of them, confirm the phylogenetic relations in this family. The specimens with a less complex pollen morphology were the trees or shrubs from the genera Brosimum, Cepropia, Chlorophora and Coussapoa, followed by Ficus, Pourouma and Sorocea; the pollen grains from the herbaceous Dorstenia show a complex pollen morphology. The structures of the sporoderm and the apertures of the pollen grains point out the relationship between these genera and other families from the Urticales.

Acute physiological impacts of CO2 ocean sequestration on marine animals

International Nuclear Information System (INIS)

Ishimatsu, A.; Hayashi, M.; Lee, K.S.; Murata, K.; Kumagai, E.

2005-01-01

The biological impacts of ocean carbon dioxide (CO 2 ) sequestration must be carefully considered before it is implemented as a mitigation strategy. This paper presented details of a study investigating the effects of high CO 2 concentrations on marine fish, lobster, and octopus. The influence of water temperature on the physiological effects of CO 2 was also discussed. In the first part of the study, eggs and larvae of red seabream were exposed to both CO 2 and HCI-acidified seawater at identical pH levels. Seabream in the CO 2 group showed a much higher mortality rate than fish in the HCI group. Other tests showed that Japanese Flounder died after complete recovery of pH in seawater equilibrated with 5 per cent CO 2 . Cardiac output was rapidly depressed in Yellowtail fish without significant changes in blood oxygen concentrations. Lower temperatures resulted in higher mortality and delayed pH recovery during hypercapnia in all fish. Western rock lobsters were the most tolerant to CO 2 among all species tested. The recovery of hemolymph pH was complete at exposure to CO 2 concentrations of 1 per cent. Changes in hemolymph bicarbonate concentrations indicated that acid-based regulatory mechanisms differed between fish and lobsters. Mortality rates for octopus were significant at CO 2 concentrations of 1 per cent. The results of all tests showed that aquatic animals are more susceptible to increases in ambient CO 2 levels than terrestrial animals. It was concluded that even slight elevations in CO 2 concentration levels adversely affected physiological functioning in the tested species. It was concluded that CO 2 sequestration in deeper, colder waters will have a more pronounced effect on aquatic animals due to the interactions between CO 2 and lower temperatures, as well as the fact that most deep-sea fish are less tolerant to environmental perturbations. 3 refs., 1 tab., 3 figs

Acute physiological impacts of CO{sub 2} ocean sequestration on marine animals

Energy Technology Data Exchange (ETDEWEB)

Ishimatsu, A.; Hayashi, M.; Lee, K.S.; Murata, K.; Kumagai, E. [Nagasaki Univ., Nagasaki (Japan). Marine Research Inst.; Kikkawa, T. [Marine Ecology Research Inst., Chiba (Japan). Central Laboratory; Kita, J. [Research Inst. of Innovative Technology for the Earth, Kyoto (Japan)

2005-07-01

The biological impacts of ocean carbon dioxide (CO{sub 2}) sequestration must be carefully considered before it is implemented as a mitigation strategy. This paper presented details of a study investigating the effects of high CO{sub 2} concentrations on marine fish, lobster, and octopus. The influence of water temperature on the physiological effects of CO{sub 2} was also discussed. In the first part of the study, eggs and larvae of red seabream were exposed to both CO{sub 2} and HCI-acidified seawater at identical pH levels. Seabream in the CO{sub 2} group showed a much higher mortality rate than fish in the HCI group. Other tests showed that Japanese Flounder died after complete recovery of pH in seawater equilibrated with 5 per cent CO{sub 2}. Cardiac output was rapidly depressed in Yellowtail fish without significant changes in blood oxygen concentrations. Lower temperatures resulted in higher mortality and delayed pH recovery during hypercapnia in all fish. Western rock lobsters were the most tolerant to CO{sub 2} among all species tested. The recovery of hemolymph pH was complete at exposure to CO{sub 2} concentrations of 1 per cent. Changes in hemolymph bicarbonate concentrations indicated that acid-based regulatory mechanisms differed between fish and lobsters. Mortality rates for octopus were significant at CO{sub 2} concentrations of 1 per cent. The results of all tests showed that aquatic animals are more susceptible to increases in ambient CO{sub 2} levels than terrestrial animals. It was concluded that even slight elevations in CO{sub 2} concentration levels adversely affected physiological functioning in the tested species. It was concluded that CO{sub 2} sequestration in deeper, colder waters will have a more pronounced effect on aquatic animals due to the interactions between CO{sub 2} and lower temperatures, as well as the fact that most deep-sea fish are less tolerant to environmental perturbations. 3 refs., 1 tab., 3 figs.

Silencing of RpATG6 impaired the yolk accumulation and the biogenesis of the yolk organelles in the insect vector R. prolixus.

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Priscila H Vieira

2018-05-01

Full Text Available In oviparous animals, the egg yolk is synthesized by the mother in a major metabolic challenge, where the different yolk components are secreted to the hemolymph and delivered to the oocytes mostly by endocytosis. The yolk macromolecules are then stored in a wide range of endocytic-originated vesicles which are collectively referred to as yolk organelles and occupy most of the mature oocytes cytoplasm. After fertilization, the contents of these organelles are degraded in a regulated manner to supply the embryo cells with fundamental molecules for de novo synthesis. Yolk accumulation and its regulated degradation are therefore crucial for successful development, however, most of the molecular mechanisms involved in the biogenesis, sorting and degradation of targeted yolk organelles are still poorly understood. ATG6 is part of two PI3P-kinase complexes that can regulate the recruitment of the endocytic or the autophagy machineries. Here, we investigate the role of RpATG6 in the endocytosis of the yolk macromolecules and in the biogenesis of the yolk organelles in the insect vector Rhodnius prolixus. We found that vitellogenic females express high levels of RpATG6 in the ovaries, when compared to the levels detected in the midgut and fat body. RNAi silencing of RpATG6 resulted in yolk proteins accumulated in the vitellogenic hemolymph, as a consequence of poor uptake by the oocytes. Accordingly, the silenced oocytes are unviable, white (contrasting to the control pink oocytes, smaller (62% of the control oocyte volume and accumulate only 40% of the yolk proteins, 80% of the TAG and 50% of the polymer polyphosphate quantified in control oocytes. The cortex of silenced oocytes present atypical smaller vesicles indicating that the yolk organelles were not properly formed and/or sorted, which was supported by the lack of endocytic vesicles near the plasma membrane of silenced oocytes as seen by TEM. Altogether, we found that RpATG6 is central for the

JAK/STAT signaling pathway-mediated immune response in silkworm (Bombyx mori) challenged by Beauveria bassiana.

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Geng, Tao; Lv, Ding-Ding; Huang, Yu-Xia; Hou, Cheng-Xiang; Qin, Guang-Xing; Guo, Xi-Jie

2016-12-20

Innate immunity was critical in insects defensive system and able to be induced by Janus kinase/signal transducer and activator of transcription cascade transduction (JAK/STAT) signaling pathway. Currently, it had been identified many JAK/STAT signaling pathway-related genes in silkworm, but little function was known on insect innate immunity. To explore the roles of JAK/STAT pathway in antifungal immune response in silkworm (Bombyx mori) against Beauveria bassiana infection, the expression patterns of B. mori C-type lectin 5 (BmCTL5) and genes encoding 6 components of JAK/STAT signaling pathway in silkworm challenged by B. bassiana were analyzed using quantitative real time PCR. Meanwhile the activation of JAK/STAT signaling pathway by various pathogenic micro-organisms and the affect of JAK/STAT signaling pathway inhibitors on antifungal activity in silkworm hemolymph was also detected. Moreover, RNAi assay of BmCTL5 and the affect on expression levels of signaling factors were also analyzed. We found that JAK/STAT pathway could be obviously activated in silkworm challenged with B. bassiana and had no response to bacteria and B. mori cytoplasmic polyhedrosis virus (BmCPV). However, the temporal expression patterns of JAK/STAT signaling pathway related genes were significantly different. B. mori downstream receptor kinase (BmDRK) might be a positive regulator of JAK/STAT signaling pathway in silkworm against B. bassiana infection. Moreover, antifungal activity assay showed that the suppression of JAK/STAT signaling pathway by inhibitors could significantly inhibit the antifungal activity in hemolymph and resulted in increased sensitivity of silkworm to B. bassiana infection, indicating that JAK/STAT signaling pathway might be involved in the synthesis and secretion of antifungal substances. The results of RNAi assays suggested that BmCTL5 might be one pattern recognition receptors for JAK/STAT signaling pathway in silkworm. These findings yield insights for better

The innate immune and systemic response in honey bees to a bacterial pathogen, Paenibacillus larvae

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Foster Leonard J

2009-08-01

Full Text Available Abstract Background There is a major paradox in our understanding of honey bee immunity: the high population density in a bee colony implies a high rate of disease transmission among individuals, yet bees are predicted to express only two-thirds as many immunity genes as solitary insects, e.g., mosquito or fruit fly. This suggests that the immune response in bees is subdued in favor of social immunity, yet some specific immune factors are up-regulated in response to infection. To explore the response to infection more broadly, we employ mass spectrometry-based proteomics in a quantitative analysis of honey bee larvae infected with the bacterium Paenibacillus larvae. Newly-eclosed bee larvae, in the second stage of their life cycle, are susceptible to this infection, but become progressively more resistant with age. We used this host-pathogen system to probe not only the role of the immune system in responding to a highly evolved infection, but also what other mechanisms might be employed in response to infection. Results Using quantitative proteomics, we compared the hemolymph (insect blood of five-day old healthy and infected honey bee larvae and found a strong up-regulation of some metabolic enzymes and chaperones, while royal jelly (food and energy storage proteins were down-regulated. We also observed increased levels of the immune factors prophenoloxidase (proPO, lysozyme and the antimicrobial peptide hymenoptaecin. Furthermore, mass spectrometry evidence suggests that healthy larvae have significant levels of catalytically inactive proPO in the hemolymph that is proteolytically activated upon infection. Phenoloxidase (PO enzyme activity was undetectable in one or two-day-old larvae and increased dramatically thereafter, paralleling very closely the age-related ability of larvae to resist infection. Conclusion We propose a model for the host response to infection where energy stores and metabolic enzymes are regulated in concert with direct

Behavioral Responses of Concholepas concholepas (Bruguière, 1789) Larvae to Natural and Artificial Settlement Cues and Microbial Films.

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Rodriguez, S R; Riquelme, C; Campos, E O; Chavez, P; Brandan, E; Inestrosa, N C

1995-12-01

The behavioral responses of veliger larvae of the gastropod Concholepas concholepas were studied in the presence of different natural and artificial settlement cues and microbial films. Early pre-competent larvae stopped swimming, sank (due to ciliary arrests, retraction of the velum into the shell, or both), and remained inactive on the substratum when exposed to conspecific mucus and hemolymph. In both cases the effect was time-dependent and the number of larvae showing these behaviors decreased over time. Larvae exposed to NH4Cl (ammonium ion) showed a similar time- and dose-dependent response. A positive and time-dependent response was also observed when larvae were exposed to different extracellular matrix (ECM) components (i.e., collagen, gelatin, and fibronectin) and sulfated polysaccharides (i.e., carrageenan, heparin, and chondroitin sulfate). In this case the larvae remained attached to the substratum. However, the effect of sulfated polysaccharides on C. concholepas larval behavior was faster than that observed with other ECM molecules. We also studied the responses of premetamorphic C. concholepas larvae exposed to different microbial films. In chemotaxis experiments with different films, with glass as the substratum, larvae showed a significant preference for multispecific and diatoms films. When shells of C. concholepas were used as the substratum, the preference for multispecific films was clear and significant. Likewise, larvae showed velar contractions in the presence of all the films tested. Larvae exposed to multispecific films and to the microalga Prasinocladus marinus showed an increased ciliar movement. The finding that mucus and hemolymph of conspecific adults and ECM molecules (mainly sulfated polysaccharides) induce the cessation of swimming of C. concholepas larvae suggests a possible role for cell-surface receptors in mediating the larval response of marine organisms. Likewise, the positive chemotaxis responses of C. concholepas larvae to

The IGFBP7 homolog Imp-L2 promotes insulin signaling in distinct neurons of the Drosophila brain.

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Bader, R; Sarraf-Zadeh, L; Peters, M; Moderau, N; Stocker, H; Köhler, K; Pankratz, M J; Hafen, E

2013-06-15

In Drosophila, Insulin-like peptide 2 (Dilp-2) is expressed by insulin-producing cells in the brain, and is secreted into the hemolymph to activate insulin signaling systemically. Within the brain, however, a more local activation of insulin signaling may be required to couple behavioral and physiological traits to nutritional inputs. We show that a small subset of neurons in the larval brain has high Dilp-2-mediated insulin signaling activity. This local insulin signaling activation is accompanied by selective Dilp-2 uptake and depends on the expression of the Imaginal morphogenesis protein-late 2 (Imp-L2) in the target neurons. We suggest that Imp-L2 acts as a licensing factor for neuronal IIS activation through Dilp-2 to further increase the precision of insulin activity in the brain.

The dipteran parasitoid Exorista bombycis induces pro- and anti-oxidative reactions in the silkworm Bombyx mori: Enzymatic and genetic analysis.

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Makwana, Pooja; Pradeep, Appukuttan Nair R; Hungund, Shambhavi P; Ponnuvel, Kangayam M; Trivedy, Kanika

2017-02-01

Hymenopteran parasitoids inject various factors including polydnaviruses along with their eggs into their host insects that suppress host immunity reactions to the eggs and larvae. Less is known about the mechanisms evolved in dipteran parasitoids that suppress host immunity. Here we report that the dipteran, Exorista bombycis, parasitization leads to pro-oxidative reactions and activation of anti-oxidative enzymes in the silkworm Bombyx mori larva. We recorded increased activity of oxidase, superoxide dismutase, thioredoxin peroxidase, catalase, glutathione-S-transferase (GST), and peroxidases in the hemolymph plasma, hemocytes, and fat body collected from B. mori after E. bombycis parasitization. Microarray and qPCR showed differential expression of genes encoding pro- and anti-oxidant enzymes in the hemocytes. The significance of this work lies in increased understanding of dipteran parasitoid biology. © 2017 Wiley Periodicals, Inc.

Evidence of spotted fever group rickettsiae in state of Rio de Janeiro, Brazil

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ROZENTAL Tatiana

2002-01-01

Full Text Available Ticks were obtained from dogs from February to September of 1999 at weekly intervals, in the County of Piraí, State of Rio de Janeiro. Four hundred seventy four ixodids were taxonomically identified, 103 Amblyomma cajennense, seven Amblyomma ovale, 209 Rhipicephalus sanguineus, and 155 Amblyomma sp. An hemolymph test associated with Giemsa's stain revealed two specimens in 163 ticks tested (R. sanguineus and Amblyomma sp, containing rickettsia-like organisms. Direct immunofluorescence verified the presence of spotted fever group rickettsia in one specimen of R. sanguineus. Considering the limited information on rickettsiosis in Brazil, principally in relation to the vectors involved in perpetuating it in foci, these preliminary results give us an idea on the importance of infection in ticks, allowing to expand our knowledge on this zoonosis.

Methyl farnesoate couples environmental changes to testicular development in the crab Carcinus maenas.

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Nagaraju, G P C; Borst, D W

2008-09-01

Carcinus maenas males have two major color phases. Green-phase males molt frequently and tend to live in brackish estuaries during the summer. After becoming red-phase males, they molt infrequently, have higher mating success, and live in cooler, deeper water. We found profound differences between these two phases in the way salinity and temperature affect hemolymph levels of methyl farnesoate (MF), a hormone that affects crustacean reproduction. Few green-phase males (salinity. MF levels of green-phase males were increased by injecting MF, by ESA, or by exposure to 5 ppt SW at 18 degrees C. The testicular index of these treated animals nearly doubled after two weeks. Our results strongly suggest that environmental conditions such as temperature and salinity, affect testicular development in this crab by changing its MF levels.

Humoral and Haemocytic Responses of Litopenaeus vannamei to Cd Exposure

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Juan C. Bautista-Covarrubias

2014-01-01

Full Text Available White shrimp, Litopenaeus vannamei, subadults were exposed to four dilutions of the 96 h cadmium LC50 reported for postlarvae (PL12 of this species, and the effects were evaluated after 5, 48, and 96 h of exposure. While treatments did not affect survival and hemolymph clotting time increased with time, but not as a response to Cd exposure, the intensity of other responses was related to concentration, to time of exposure, and to their interaction. Hemocyanin decreased with time in all metal concentrations but increased in the control treatment, and an almost similar trend was observed with hemocyte numbers. As an initial response, phenoloxidase activity decreased with all metal concentrations, but it increased later to values similar or higher than the control treatment.

Polliniferous plants aud foraging strategles Of Apis mellifera (Hyínenoptera: Apidae in the Yucatán Peninsula, Mexico

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Rogel Villanueva-G

2002-12-01

Full Text Available A study of the most important polliniferous plants for European and Africanized honeybees (Apis mellifera L. was made in Quintana Roo state. Comparisons were made between the plants visited by both bee types in order to determine whether there were qualitative or quantitative differences in their choice of plant species. Also some foraging strategies of the honeybees were analysed. Pollen from pollen load samples was acetolysed and mounted on slides. Subsequently the polien grains were identified, counted and photographed. A total of 206 pollen load samples were collected at Palmas and St. Teresa during two years. The most frequent species in the ponen load samples from European and Africanized honeyhees were Cecropia peltata, Metopium brownei, Lonchocarpus sp. 2, Viguiera dentata, Eragrostis sp. 1, Bursera simaruba and Eupatorium albicaule. Both types of honey bees show a high reliance on pollen from only a few species, the first five named above comprised around 50% of all the mean percentage frequencies. Families that contributed with the largest number of polien species were Fabaceae, Asteraceac, Boraginaceae, Convolvulaecae, Euphorbiaceae, Sapindaceae, Poaceac, Myrtaceae, Sapotaceae and Tiliaceae. C. peltata, Trema micrantha, B. simaruba, Eugenia sp. 1, Thouinia canesceras, Pouteria sp. 1, Mimosa bahamensis and V. dentata, were the pollen species with the largest percentages of oceurrence in both European and Africanized bee pollen load samples, and also represent a "long-term" food resources during the year.Un estudio de las plantas poliníferas más importantes para las abejas europeas y africanizadas (Apis melifera L. se realizó en el estado de Quintana Roo. Se hicieron comparaciones entre plantas visitadas por ambos tipos de abejas, con el objetivo de determinar si hay diferencias cualitativas o cuantitativas en la elección de la especie de planta. Adicionalmente, se analizaron algunas estrategias de forrajeo de las abejas. Muestras

Revision of the Neotropical spider genus Gephyroctenus (Araneae: Ctenidae: Calocteninae Revisão do gênero de aranhas neotropicais Gephyroctenus Mello-Leitão (Araneae: Ctenidae: Calocteninae

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Daniele Polotow

2008-12-01

Full Text Available Gephyroctenus Mello-Leitão, 1936 (type species G. philodromoides Mello-Leitão, 1936 is revised. Three species formerly described in this genus are synonymized with species from other genera: G. kolosvaryi Caporiacco, 1947, with Acanthoctenus spinipes Keyserling, 1877, G. parvus Caporiacco, 1947, with Phymatoctenus comosus Simon, 1897, and G. vachoni Caporiacco, 1955, with Caloctenus gracilitarsis Simon, 1897. Eight new species are described: G. portovelho sp. nov., from the States of Rondônia and Amazonas, Brazil; G. divisor sp. nov. and G. acre sp. nov., from the State of Acre, Brazil; G. atininga sp. nov., G. esteio sp. nov. and G. mapia sp. nov., from the State of Amazonas, Brazil; G. juruti sp. nov., from the Department of Loreto, Peru and the State of Pará, Brazil; G. panguana sp. nov., from the Department of Huanuco, Peru. The genus can be distinguished by the presence of a cymbial retrolateral groove, retrolateral origin of embolus, embolus long and thin, median apophysis with a subdistal hook, and hyaline projection close to the embolus base in the male palp and by the fused median and lateral fields in a single epigynal plate, copulatory opening located dorsally in an atrium, and elongated copulatory ducts surrounding the spermathecae in the female epigynum. Field observations on the hunting behavior on ants in trumpet trees (Cecropia are provided for two species, G. philodromoides and G. mapia sp. nov.Gephyroctenus Mello-Leitão, 1936 (espécie-tipo G. philodromoides Mello-Leitão, 1936, é revisado. Três espécies descritas neste gênero são sinonimizadas com espécies de outros gêneros: G. kolosvaryi Caporiacco, 1947, com Acanthoctenus spinipes Keyserling, 1877, G. parvus Caporiacco, 1947, com Phymatoctenus comosus Simon, 1897, e G. vachoni Caporiacco, 1955, com Caloctenus gracilitarsis Simon, 1897. Oito novas espécies são descritas: G. portovelho sp. nov., dos Estados de Rondônia e Amazonas, Brasil; G. divisor sp. nov. e G

Atrial granular cells of the snail Achatina fulica release proteins into hemolymph after stimulation of the heart nerve.

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Shabelnikov, Sergej V; Bystrova, Olga A; Ivanov, Vadim A; Margulis, Boris A; Martynova, Marina

2009-10-01

The atrium of the gastropod mollusc Achatina fulica receives rich innervation and contains numerous granular cells (GCs). We studied the atrial innervation and discovered that axon profiles typical in appearance of peptidergic neurons form close unspecialized membrane contacts with GCs. Then, we investigated, at both morphological and biochemical levels, the effect of electrical stimulation of the heart nerve on GCs of Achatina heart perfused in situ. The ultrastructural study demonstrated changes in granule morphology consistent with secretion. These events included alteration of granule content, intracellular granule fusion and formation of complex degranulation channels, within which the granule matrix solubilized. It was shown that electrical stimulation resulted in a significant increase of the total protein concentration in the perfusate. Furthermore, SDS-PAGE analysis of the perfusate revealed three new proteins with molecular masses of 16, 22, and 57 kDa. Affinity-purified polyclonal antibodies against the 16 kDa protein were obtained; the whole-mount immunofluorescence technique revealed the presence of this protein in the granules of atrial GCs. In GCs of the stimulated atrium, a progressive loss of their granular content was observed. The results suggest that the central nervous system can modulate the secretory activity of the atrial GCs through non-synaptic pathways.

Tropical Atlantic marine macroalgae with bioactivity against virulent and antibiotic resistant Vibrio

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Giselle Cristina Silva

2013-03-01

Full Text Available The antibacterial activity of ethanol, methanol, hexane and acetone-based extracts of the macroalgae Padina gymnospora (PG, Hypnea musciformes (HM, Ulva fasciata (UF and Caulerpa prolifera (CP was investigated. The disk diffusion method was used to evaluate the algae antimicrobial effect against standard strains of Vibrio parahaemolyticus, Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa and Salmonella enterica and five virulent antibiotic-resistant strains of V. brasiliensis, V. xuii and V. navarrensis (isolated from the hemolymph of Litopenaeus vannamei. Ethanol extracts of PG and HM inhibited all Vibrio strains. E. coli and P. aeruginosa were only susceptible to ethanol extracts of PG. Among the methanol extracts, only UF was bioactive, inhibiting V. navarrensis. The observed inhibitory effect of ethanol extracts of PG, HM and UF against virulent antibiotic-resistant bacteria suggests these macroalgal species constitute a potential source of bioactive compounds.

Caracterização dos tipos celulares presentes na hemolinfa de larvas e ninfas de Rhipicephalus sanguineus (Latreille (Ixodoidea, Ixodidae em diferentes estados nutricionais Characterization of the celular types present in the Haemolymph of larvae and nimphs of Rhipicephalus sanguineus (Latreille (Ixodoidea, Ixodidae in different nutritional stages

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Maria Elisa Carneiro

1996-01-01

Full Text Available With the purpose of characterize morphologically the hemocytes of larvae and nymphs of Rhipicephalus sanguineus on different nutritional phases, it was obtained samples of haemolymph were obtained by sectioning the forelegs and collecting the drop there formed. After dried, the samples were fixed by methanol and stained by Giemsa. Microscopical observation resulted in the characterization of five basic cellular types: prohemocytes, plasmatocytes, granulocytes, spherulocytes and oenocytoids. Moreover, undefined cell types, whose morphological patterns didn't have correlation with the characteristics cited for hemocytes, were found in low frequency. The change in the relative composition of the haemolimph was characterized by decrease of the number of granulocytes and greater variabilily of the cell types present in the hemolymph as the tick evolved. This fact may be linked to the alterations that these cells suffer along the development of the tick.

Moessbauer spectroscopic studies of iron-storage proteins

Energy Technology Data Exchange (ETDEWEB)

St. Pierre, T.G.

1986-01-01

/sup 57/Fe Moessbauer spectroscopy was used to study iron storage proteins. Various cryostats and a superconducting magnet were used to obtain sample environment temperatures from 1.3 to 200K and applied magnetic fields of up to 10T. The Moessbauer spectra of ferritins isolated from iron-overloaded human spleen, limpet (Patella vulgata), giant limpet (Patella laticostata) and chiton (Clavarizona hirtosa) hemolymph, and bacterial (Pseudomonas aeruginosa) cells are used to gain information on the magnetic ordering- and superparamagnetic transition temperatures of the microcrystalline cores of the proteins. Investigations were made about the cause of the difference in the magnetic anisotropy constants of the cores of iron-overloaded human spleen ferritin and hemosiderin. Livers taken from an iron-overloaded hornbill and artificially iron-loaded rats showed no component with a superparamagnetic transition temperature approaching that of the human spleen hemosiderin.

Drosophila insulin release is triggered by adipose Stunted ligand to brain Methuselah receptor.

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Delanoue, Renald; Meschi, Eleonora; Agrawal, Neha; Mauri, Alessandra; Tsatskis, Yonit; McNeill, Helen; Léopold, Pierre

2016-09-30

Animals adapt their growth rate and body size to available nutrients by a general modulation of insulin-insulin-like growth factor signaling. In Drosophila, dietary amino acids promote the release in the hemolymph of brain insulin-like peptides (Dilps), which in turn activate systemic organ growth. Dilp secretion by insulin-producing cells involves a relay through unknown cytokines produced by fat cells. Here, we identify Methuselah (Mth) as a secretin-incretin receptor subfamily member required in the insulin-producing cells for proper nutrient coupling. We further show, using genetic and ex vivo organ culture experiments, that the Mth ligand Stunted (Sun) is a circulating insulinotropic peptide produced by fat cells. Therefore, Sun and Mth define a new cross-organ circuitry that modulates physiological insulin levels in response to nutrients. Copyright © 2016, American Association for the Advancement of Science.

Females are the brighter sex: Differences in external fluorescence across sexes and life stages of a crab spider.

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Brandt, Erin E; Masta, Susan E

2017-01-01

Fluorescence is increasingly recognized to be widespread in nature. In particular, some arachnids fluoresce externally, and in spiders the hemolymph fluoresces. In this study, we examined the external fluorescence and the fluorophores of different sexes and life stages of the crab spider Misumena vatia (Clerk 1757), a sit-and-wait predator that feeds on insects as they visit flowers. We designed novel instrumentation to measure external fluorescence in whole specimens. We found that although males and females possess internal fluorophores with similar properties, the external expression of fluorescence varies across sexes and life stages. Spiders fluoresce brightly as immatures. Females maintain their brightness to adulthood, whereas males become increasingly dim as they mature. We suggest that external fluorescence likely contributes to visual signaling in these animals, and that it differs between the sexes as a result of differences in foraging ecology and behavior.

Biogenic Amines in Insect Antennae

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Marianna I. Zhukovskaya

2017-06-01

Full Text Available Insect antenna is a multisensory organ, each modality of which can be modulated by biogenic amines. Octopamine (OA and its metabolic precursor tyramine (TA affect activity of antennal olfactory receptor neurons. There is some evidence that dopamine (DA modulates gustatory neurons. Serotonin can serve as a neurotransmitter in some afferent mechanosensory neurons and both as a neurotransmitter and neurohormone in efferent fibers targeted at the antennal vessel and mechanosensory organs. As a neurohormone, serotonin affects the generation of the transepithelial potential by sensillar accessory cells. Other possible targets of biogenic amines in insect antennae are hygro- and thermosensory neurons and epithelial cells. We suggest that the insect antenna is partially autonomous in the sense that biologically active substances entering its hemolymph may exert their effects and be cleared from this compartment without affecting other body parts.

Arboreal Ants Use the “Velcro® Principle” to Capture Very Large Prey

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Dejean, Alain; Leroy, Céline; Corbara, Bruno; Roux, Olivier; Céréghino, Régis; Orivel, Jérôme; Boulay, Raphaël

2010-01-01

Plant-ants live in a mutualistic association with host plants known as “myrmecophytes” that provide them with a nesting place and sometimes with extra-floral nectar (EFN) and/or food bodies (FBs); the ants can also attend sap-sucking Hemiptera for their honeydew. In return, plant-ants, like most other arboreal ants, protect their host plants from defoliators. To satisfy their nitrogen requirements, however, some have optimized their ability to capture prey in the restricted environment represented by the crowns of trees by using elaborate hunting techniques. In this study, we investigated the predatory behavior of the ant Azteca andreae which is associated with the myrmecophyte Cecropia obtusa. We noted that up to 8350 ant workers per tree hide side-by-side beneath the leaf margins of their host plant with their mandibles open, waiting for insects to alight. The latter are immediately seized by their extremities, and then spread-eagled; nestmates are recruited to help stretch, carve up and transport prey. This group ambush hunting technique is particularly effective when the underside of the leaves is downy, as is the case for C. obtusa. In this case, the hook-shaped claws of the A. andreae workers and the velvet-like structure of the underside of the leaves combine to act like natural Velcro® that is reinforced by the group ambush strategy of the workers, allowing them to capture prey of up to 13,350 times the mean weight of a single worker. PMID:20593032

Biodiversidad florística del Jardín Botánico de la Universidad Nacional de Ucayali

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Víctor Araujo Abanto

2015-12-01

Full Text Available El objetivo fue la clasificación taxonómica y descripciones botánicas de 36 especies (23 familias y 33 géneros, árboles, arbustos, lianas y hierbas en el Jardín Botánico y Campus de la Universidad Nacional de Ucayali. De las 36 especies, 6 son ornamentales: Sanchezia tigrina, Crinum augustum, Sansevieria trifasciata, Dracaena fragans, Costus guanaiensis y Vitex cymosa; Rhynchosia phaseoloides cuyas semillas son cotizadas como artesanal; 7 podrían tener uso ornamental: Tabernaemontana heteropylla, Sparattosperma leucanthum, Cayaponia ophthalmica, Gurania eriantha, Gurania spinulosa, Stigmaphyllon florosum y Capirona decorticans; 3 son medicinales: Euphorbia umbellata (tratamiento del cáncer y otros males, Uncaria guianensis (refuerza el sistema inmunológico y Pilocarpus peruvianus contiene Pilocarpina de uso oftalmológico; Pourouma guianensis tiene fuerte olor a mentol; 3 con frutos alimenticios Tamarindus indica, Bunchosia armeniaca y Physalis angulata, conteniendo este último alto contenido en pro vitamina A; 10 son maderables: Dipteryx odorata, Vitex cymosa, Tectona grandis, Ocotea bofo, Perebea xanthochyma, Virola pavonis, Calycophyllum spruceanum, Capirona decorticans, Guazuma ulmifolia y Bambusa arundinacea; Pueraria phaseoloides usado como forraje, en el Jardín Botánico está como maleza al igual que Davilla nítida y Urena lobata, que podría servir como ornamental, su corteza tiene propiedades similares al yute verdadero; Cecropia membranácea, C. concolor, C. sciadophylla y Gouania lupuloides que pueden servir como alimento de la fauna silvestre.

Arboreal ants use the "Velcro(R principle" to capture very large prey.

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Alain Dejean

2010-06-01

Full Text Available Plant-ants live in a mutualistic association with host plants known as "myrmecophytes" that provide them with a nesting place and sometimes with extra-floral nectar (EFN and/or food bodies (FBs; the ants can also attend sap-sucking Hemiptera for their honeydew. In return, plant-ants, like most other arboreal ants, protect their host plants from defoliators. To satisfy their nitrogen requirements, however, some have optimized their ability to capture prey in the restricted environment represented by the crowns of trees by using elaborate hunting techniques. In this study, we investigated the predatory behavior of the ant Azteca andreae which is associated with the myrmecophyte Cecropia obtusa. We noted that up to 8350 ant workers per tree hide side-by-side beneath the leaf margins of their host plant with their mandibles open, waiting for insects to alight. The latter are immediately seized by their extremities, and then spread-eagled; nestmates are recruited to help stretch, carve up and transport prey. This group ambush hunting technique is particularly effective when the underside of the leaves is downy, as is the case for C. obtusa. In this case, the hook-shaped claws of the A. andreae workers and the velvet-like structure of the underside of the leaves combine to act like natural Velcro that is reinforced by the group ambush strategy of the workers, allowing them to capture prey of up to 13,350 times the mean weight of a single worker.

Domoic acid toxicokinetics in Dungeness crabs: New insights into mechanisms that regulate bioaccumulation

Energy Technology Data Exchange (ETDEWEB)

Schultz, Irvin R., E-mail: ir_schultz@pnl.gov; Skillman, Ann; Sloan-Evans, Siobhan; Woodruff, Dana

2013-09-15

Highlights: •Domoic acid is retained in the hepatopancreas after oral dosing but not after intravascular dosing. •Localization of domoic acid in the hepatopancreas is extracellular, likely in the lumen of tubules. •Isolated hepatopancreas cells do not absorb domoic acid from culture media. •Domoic acid is eliminated through urinary excretion. -- Abstract: Domoic acid (DA) is an excitatory neurotoxic amino acid produced by several marine algal species and is the causative agent of amnesic shellfish poisoning. Profound differences in the toxicokinetics of DA have been identified in a wide variety of shellfish. We characterized the toxicokinetics of DA in Dungeness crabs (Metacarcinus magister) after oral and intravascular dosing (IV) using a variety of doses ranging from 0.1 to 20 mg/kg. After a 1 mg/kg oral dose, DA disappeared from the foregut within 2 h and largely accumulated in the hepatopancreas, with hemolymph and other tissues having 100–1000 times lower concentrations. After IV dosing, hemolymph concentrations of DA were unexpectedly high and toxicokinetic analysis indicated the steady-state volume of distribution (V{sub ss}) was 123–197 ml/kg, which is well below the hemolymph volume of 350 ml/kg for crabs. This indicated only limited extravascular distribution of DA was occurring after IV injection, which is surprising considering the capacity of the hepatopancreas to sequester DA after oral dosing. Additional studies measured the partitioning of DA in hepatopancreas cellular and subcellular fractions. The subcellular distribution of DA was primarily associated with the S8 fraction and could be filtered through a 30,000 MW cut-off filter, indicating DA was not appreciably bound to macromolecules. Interestingly, very little (<0.4%) of the total hepatopancreas DA tissue content was associated with the cellular fraction isolated after dissociation and separation from tissue fragments. The in vivo and in vitro results led us to hypothesize that DA

Differential expression of genes encoding anti-oxidant enzymes in Sydney rock oysters, Saccostrea glomerata (Gould) selected for disease resistance.

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Green, Timothy J; Dixon, Tom J; Devic, Emilie; Adlard, Robert D; Barnes, Andrew C

2009-05-01

Sydney rock oysters (Saccostrea glomerata) selectively bred for disease resistance (R) and wild-caught control oysters (W) were exposed to a field infection of disseminating neoplasia. Cumulative mortality of W oysters (31.7%) was significantly greater than R oysters (0.0%) over the 118 days of the experiment. In an attempt to understand the biochemical and molecular pathways involved in disease resistance, differentially expressed sequence tags (ESTs) between R and W S. glomerata hemocytes were identified using the PCR technique, suppression subtractive hybridisation (SSH). Sequencing of 300 clones from two SSH libraries revealed 183 distinct sequences of which 113 shared high similarity to sequences in the public databases. Putative function could be assigned to 64 of the sequences. Expression of nine ESTs homologous to genes previously shown to be involved in bivalve immunity was further studied using quantitative reverse-transcriptase PCR (qRT-PCR). The base-line expression of an extracellular superoxide dismutase (ecSOD) and a small heat shock protein (sHsP) were significantly increased, whilst peroxiredoxin 6 (Prx6) and interferon inhibiting cytokine factor (IK) were significantly decreased in R oysters. From these results it was hypothesised that R oysters would be able to generate the anti-parasitic compound, hydrogen peroxide (H(2)O(2)) faster and to higher concentrations during respiratory burst due to the differential expression of genes for the two anti-oxidant enzymes of ecSOD and Prx6. To investigate this hypothesis, protein extracts from hemolymph were analysed for oxidative burst enzyme activity. Analysis of the cell free hemolymph proteins separated by native-polyacrylamide gel electrophoresis (PAGE) failed to detect true superoxide dismutase (SOD) activity by assaying dismutation of superoxide anion in zymograms. However, the ecSOD enzyme appears to generate hydrogen peroxide, presumably via another process, which is yet to be elucidated. This

Immunological responses of the mangrove oysters Crassostrea gasar naturally infected by Perkinsus sp. in the Mamanguape Estuary, Paraíba state (Northeastern, Brazil).

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Queiroga, Fernando Ramos; Marques-Santos, Luis Fernando; Hégaret, Hélène; Soudant, Philippe; Farias, Natanael Dantas; Schlindwein, Aline Daiane; Mirella da Silva, Patricia

2013-08-01

Perkinsus genus includes protozoan parasites of marine mollusks, especially bivalves. In the last four years, this parasite has been detected in mangrove oysters Crassostrea rhizophorae and Crassostrea gasar from the Northeastern region of Brazil. Hemocytes are the key cells of the oyster immune system, being responsible for a variety of cellular and humoral reactions, such as phagocytosis, encapsulation and the release of several effector molecules that control the invasion and proliferation of microorganisms. In Brazil, there is little information on perkinsosis and none on the immune responses of native oysters' species against Perkinsus spp. The objective of this study was to determine the effects of natural infection by Perkinsus sp. on the immunological parameters of mangrove oysters C. gasar cultured in the Mamanguape River Estuary (Paraíba, Brazil). Adults oysters (N = 40/month) were sampled in December 2011, March, May, August and October 2012. Gills were removed and used to determine the presence and intensity of the Perkinsus sp. infection, according to a scale of four levels (1-4), using the Ray's fluid thioglycollate medium assay. Immunological parameters were measured in hemolymph samples by flow cytometry, including: total hemocyte count (THC), differential hemocyte count (DHC), cell mortality, phagocytic capacity, and production of Reactive Oxygen Species (ROS). The plasma was used to determine the hemagglutination activity. The results showed the occurrence of Perkinsus sp. with the highest mean prevalence (93.3%) seen so far in oyster populations in Brazil. Despite that, no oyster mortality was associated. In contrast, we observed an increase in hemocyte mortality and a suppression of two of the main defense mechanisms, phagocytosis and ROS production in infected oysters. The increase in the percentage of blast-like cells on the hemolymph, and the increase in THC in oysters heavily infected (at the maximum intensity, 4) suggest an induction of

The hemocytes of Panstrogyllus Megistus (Hemiptera: Reduviidae Os hemócitos de Panstrongylus megistus (Hemiptera: Reduviidae

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Margherita Anna Barracco

1987-09-01

Full Text Available Five hemocyte types were identified in the hemolymph of Panstrongylus megistus by phase contrast and common light microscopy using some histochemical methods. These are: Prohemocytes, small cells presenting a great nucleus/cytoplasm ratio; Plasmatocytes, the most numerous hemocytes, are polymorphic cells mainly characterized by a large amount of lysosomes; Granulocytes, hemocytes very similar to plasmatocytes which contain cytoplasmic granules and are especially rich in polysaccharides; Oenocytoids, cells presenting a small nucleus and a thick cytoplasm; they show many small round vacuoles when observed in Giemsa smears and many cytoplasmic granules under phase microscopy; Adipohemocytes, very large hemocytes, presenting many fat droplet inclusions which could correspond to free fat bodies which entered the hemolymph. Only prohemocytes and plasmatocytes can be clearly classified; all the other hemocyte types have a more ambiguous classification.Cinco tipos de hemócitos foram identificados na hemolinfa de Panstrongylus megistus através da microscopia de constraste de fase e de luz, usando alguns testes histoquímicos: Pró-hemócitos-células pequenas que mostram uma grande relação núcleocitoplasmática; Plasmotócitos-células polimporficas, que se caracterizam principalmente pela sua grande abundância em lisossomos - são os hemócitos mais numerosos; Granulócitos-células muito semelhantes aos plasmatócitos que contêm grânulos citoplasmáticos particularmente ricos em polissacarídeos; Enocitóides-hemócitos que apresentam um núcleo pequeno e cujo citoplasma basofílico revela-se muito denso e homogênio - mostram uma série de pequenos vacúolos esféricos quando observados nos esfregaços corados pelo Giemsa, mas a microscopia de fase revela uma grande quantidade de pequenos grânulo ao invés de vacúolos; Adipo-hemócitos-hemócitos muito grandes que contêm uma grande quantidade de inclusões lípicas - poderiam corresponder a

The influence of diet on the δ 13C of shell carbon in the pulmonate snail Helix aspersa

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Stott, Lowell D.

2002-02-01

The influence of diet and atmospheric CO 2 on the carbon isotope composition of shell aragonite and shell-bound organic carbon in the pulmonate snail Helix aspersa raised in the laboratory was investigated. Three separate groups of snails were raised on romaine lettuce (C3 plant, δ 13C=-25.8‰), corn (C4 plant, δ 13C=-10.5‰), and sour orange ( 12C-enriched C3 plant, δ 13C=-39.1‰). The isotopic composition of body tissues closely tracked the isotopic composition of the snail diet as demonstrated previously. However, the isotopic composition of the acid insoluble organic matrix extracted from the aragonite shells does not track diet in all groups. In snails that were fed corn the isotopic composition of the organic matrix was more negative than the body by as much as 5‰ whereas the matrix was approximately 1‰ heavier than the body tissues in snails fed a diet of C3 plant material. These results indicate that isotopic composition of the organic matrix carbon cannot be used as an isotopic substrate for paleodietary reconstructions without first determining the source of the carbon and any associated fractionations. The isotopic composition of the shell aragonite is offset from the body tissues by 12.3‰ in each of the culture groups. This offset was not influenced by the consumption of carbonate and is not attributable to the diffusion of atmospheric CO 2 into the hemolymph. The carbon isotopic composition of shell aragonite is best explained in terms of equilibrium fractionations associated with exchange between metabolic CO 2 and HCO 3 in the hemolymph and the fractionation associated with carbonate precipitation. These results differ from previous studies, based primarily on samples collected in the field, that have suggested atmospheric carbon dioxide contributes significantly to the shell δ 13C. The culture results indicate that the δ 13C of aragonite is a good recorder of the isotopic composition of the snail body tissue, and therefore a better

Physiological responses of Daphnia pulex to acid stress

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Pirow Ralph

2009-04-01

Full Text Available Abstract Background Acidity exerts a determining influence on the composition and diversity of freshwater faunas. While the physiological implications of freshwater acidification have been intensively studied in teleost fish and crayfish, much less is known about the acid-stress physiology of ecologically important groups such as cladoceran zooplankton. This study analyzed the extracellular acid-base state and CO2 partial pressure (PCO2, circulation and ventilation, as well as the respiration rate of Daphnia pulex acclimated to acidic (pH 5.5 and 6.0 and circumneutral (pH 7.8 conditions. Results D. pulex had a remarkably high extracellular pH of 8.33 and extracellular PCO2 of 0.56 kPa under normal ambient conditions (pH 7.8 and normocapnia. The hemolymph had a high bicarbonate concentration of 20.9 mM and a total buffer value of 51.5 meq L-1 pH-1. Bicarbonate covered 93% of the total buffer value. Acidic conditions induced a slight acidosis (ΔpH = 0.16–0.23, a 30–65% bicarbonate loss, and elevated systemic activities (tachycardia, hyperventilation, hypermetabolism. pH 6.0 animals partly compensated the bicarbonate loss by increasing the non-bicarbonate buffer value from 2.0 to 5.1 meq L-1 pH-1. The extracellular PCO2 of pH 5.5 animals was significantly reduced to 0.33 kPa, and these animals showed the highest tolerance to a short-term exposure to severe acid stress. Conclusion Chronic exposure to acidic conditions had a pervasive impact on Daphnia's physiology including acid-base balance, extracellular PCO2, circulation and ventilation, and energy metabolism. Compensatory changes in extracellular non-bicarbonate buffering capacity and the improved tolerance to severe acid stress indicated the activation of defense mechanisms which may result from gene-expression mediated adjustments in hemolymph buffer proteins and in epithelial properties. Mechanistic analyses of the interdependence between extracellular acid-base balance and CO2 transport

Phagocytic activities of hemocytes from the deep-sea symbiotic mussels Bathymodiolus japonicus, B. platifrons, and B. septemdierum.

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Tame, Akihiro; Yoshida, Takao; Ohishi, Kazue; Maruyama, Tadashi

2015-07-01

Deep-sea mytilid mussels harbor symbiotic bacteria in their gill epithelial cells that are horizontally or environmentally transmitted to the next generation of hosts. To understand the immune defense system in deep-sea symbiotic mussels, we examined the hemocyte populations of the symbiotic Bathymodiolus mussel species Bathymodiolus japonicus, Bathymodiolus platifrons, and Bathymodiolus septemdierum, and characterized three types of hemocytes: agranulocytes (AGs), basophilic granulocytes (BGs), and eosinophilic granulocytes (EGs). Of these, the EG cells were the largest (diameter, 8.4-10.0 μm) and had eosinophilic cytoplasm with numerous eosinophilic granules (diameter, 0.8-1.2 μm). Meanwhile, the BGs were of medium size (diameter, 6.7-8.0 μm) and contained small basophilic granules (diameter, 0.3-0.4 μm) in basophilic cytoplasm, and the AGs, the smallest of the hemocytes (diameter, 4.8-6.0 μm), had basophilic cytoplasm lacking granules. A lectin binding assay revealed that concanavalin A bound to all three hemocyte types, while wheat germ agglutinin bound exclusively to EGs and BGs. The total hemocyte population densities within the hemolymph of all three Bathymodiolus mussel species were similar (8.4-13.3 × 10(5) cells/mL), and the percentages of circulating AGs, BGs, and EGs in the hemolymph of these organisms were 44.7-48.5%, 14.3-17.6%, and 34.3-41.0%, respectively. To analyze the functional differences between these hemocytes, the phagocytic activity and post-phagocytic phagosome-lysosome fusion events were analyzed in each cell type using a fluorescent Alexa Fluor(®) 488-conjugated Escherichia coli bioparticle and a LysoTracker(®) lysosomal marker, respectively. While the AGs exhibited no phagocytic activity, both types of granulocytes were phagocytic. Of the three hemocyte types, the EGs exhibited the highest level of phagocytic activity as well as rapid phagosome-lysosome fusion, which occurred within 2 h of incubation. Meanwhile, the BGs showed

Effect of gamma irradiation on the total nitrogen and protein content in body during different stages of silkworm development

International Nuclear Information System (INIS)

Petkov, N.; Malinova, K.; Binkh, N.T.

1996-01-01

The aim was to determine the effect of gamma irradiation of eggs of silk moth in B 2 stage in doses of 1.00, 2.00 and 3.00 Gy on the changes of total nitrogen and protein content during different stages of Bombyx mori L. development. Highest levels of total nitrogen and protein were found in silk gland 14.032-14.355 mg%, followed by pupae - 7.448-8.092 and 46.550-48.906 mg%, moths after egg laying - 6.650-7.825 and 41.563-48.906 mg% and silkworm hemolymph - 6.920-6.980 and 43.250-43.625 mg%, respectively. The irradiation of eggs with 2.00 and 3,00 Gy gamma rays stimulated the increase of total nitrogen and protein content in silk gland by 6.66-7.3% compared to non-irradiated eggs of the same breed. 14 refs., 3 tabs. (author)

Motility precedes egress of malaria parasites from oocysts

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Klug, Dennis; Frischknecht, Friedrich

2017-01-01

Malaria is transmitted when an infected Anopheles mosquito deposits Plasmodium sporozoites in the skin during a bite. Sporozoites are formed within oocysts at the mosquito midgut wall and are released into the hemolymph, from where they invade the salivary glands and are subsequently transmitted to the vertebrate host. We found that a thrombospondin-repeat containing sporozoite-specific protein named thrombospondin-releated protein 1 (TRP1) is important for oocyst egress and salivary gland invasion, and hence for the transmission of malaria. We imaged the release of sporozoites from oocysts in situ, which was preceded by active motility. Parasites lacking TRP1 failed to migrate within oocysts and did not egress, suggesting that TRP1 is a vital component of the events that precede intra-oocyst motility and subsequently sporozoite egress and salivary gland invasion. DOI: http://dx.doi.org/10.7554/eLife.19157.001 PMID:28115054

Pharmacokinetics of cefovecin (Convenia) in white bamboo sharks (Chiloscyllium plagiosum) and Atlantic horseshoe crabs (Limulus polyphemus).

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Steeil, James C; Schumacher, Juergen; George, Robert H; Bulman, Frank; Baine, Katherine; Cox, Sherry

2014-06-01

Cefovecin was administered to six healthy adult white bamboo sharks (Chiloscyllium plagiosum) and six healthy adult Atlantic horseshoe crabs (Limulus polyphemus) to determine its pharmacokinetics in these species. A single dose of cefovecin at 8 mg/kg was administered subcutaneously in the epaxial region of the bamboo sharks and in the proximal articulation of the lateral leg of the horseshoe crabs. Blood and hemolymph samples were collected at various time points from bamboo sharks and Atlantic horseshoe crabs. High performance liquid chromatography was performed to determine plasma levels of cefovecin. The terminal halflife of cefovecin in Atlantic horseshoe crabs was 37.70 +/- 9.04 hr and in white bamboo sharks was 2.02 +/- 4.62 hr. Cefovecin concentrations were detected for 4 days in white bamboo sharks and for 14 days in Atlantic horseshoe crabs. No adverse effects associated with cefovecin administration were seen in either species.

Females are the brighter sex: Differences in external fluorescence across sexes and life stages of a crab spider.

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Erin E Brandt

Full Text Available Fluorescence is increasingly recognized to be widespread in nature. In particular, some arachnids fluoresce externally, and in spiders the hemolymph fluoresces. In this study, we examined the external fluorescence and the fluorophores of different sexes and life stages of the crab spider Misumena vatia (Clerk 1757, a sit-and-wait predator that feeds on insects as they visit flowers. We designed novel instrumentation to measure external fluorescence in whole specimens. We found that although males and females possess internal fluorophores with similar properties, the external expression of fluorescence varies across sexes and life stages. Spiders fluoresce brightly as immatures. Females maintain their brightness to adulthood, whereas males become increasingly dim as they mature. We suggest that external fluorescence likely contributes to visual signaling in these animals, and that it differs between the sexes as a result of differences in foraging ecology and behavior.

Flowers, fruits, and the abundance of the yellow-chevroned parakeet (Brotogeris chiriri at a gallery forest in the South Pantanal (Brazil Flores, frutos e abundância do periquito-de-asa-amarela (Brotogeris chiriri em uma mata ciliar do Pantanal Sul (Brasil

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J. Ragusa-Netto

2004-11-01

Full Text Available Parakeets usually forage for massive and ephemeral plant resources at forest canopies. Fruit pulp is widely cited as a major food resource for these birds, which often eat seeds and nectar. In this study, I assessed flower and fruit production at a gallery forest in the Pantanal flood plain (Brazil in order to evaluate the relationship between food resource production and abundance of a common parakeet, Brotogeris chiriri. Also, I evaluated the relationship between food resource production and foraging activity. Parakeet abundance varied markedly along the year, coinciding with massive episodes of flower and fleshy fruit availability. Inga vera nectar, intensely used during the latter part of dry season, was by far the most exploited food item by parakeets when they were very abundant. The nectar comprised 34% of the parakeets' diet (N = 131 feeding records at the gallery forest, while fleshy fruits made up the rest. Parakeets principally exploited fruits of Cecropia pachystachya and Ficus luschnathiana, besides palm fruits and Inga vera arils. The consistent relationship between foraging activity and parakeet abundance, as well as the coincidence between fluctuations of these parameters and availability of major food resources, suggests that food availability mostly influenced B. chiriri occurrence in the gallery forest. Furthermore, I found no evidence for gallery forest use for roosting and/or breeding, in spite of the fact that such factors usually influence local parakeet abundance.Periquitos normalmente exploram recursos massivos e efêmeros no dossel das florestas tropicais. Dentre os itens alimentares mais utilizados está a polpa de frutos, embora sementes e néctar também sejam consumidos. Neste estudo, foi avaliada a produção de flores e frutos em uma mata ciliar do Pantanal (Brasil e sua relação com a abundância do periquito Brotogeris chiriri. Além disso, avaliaram-se as relações entre a produção de flores e frutos e o

Nitrogen use strategies of seedlings from neotropical tree species of distinct successional groups.

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Oliveira, Halley Caixeta; da Silva, Ligia Maria Inocêncio; de Freitas, Letícia Dias; Debiasi, Tatiane Viegas; Marchiori, Nidia Mara; Aidar, Marcos Pereira Marinho; Bianchini, Edmilson; Pimenta, José Antonio; Stolf-Moreira, Renata

2017-05-01

Few studies have analyzed the strategies of neotropical tree seedlings for absorbing, translocating and assimilating the nitrogen. Here, we compared the nitrogen use strategies of seedlings from six tree species that are native to the Brazilian Atlantic Forest and that belong to different successional groups: Trema micrantha, Heliocarpus popayanensis and Cecropia pachystachya (pioneers), Cariniana estrellensis, Eugenia brasiliensis and Guarea kunthiana (non-pioneers). The effects of cultivating seedlings with nitrate or ammonium on the growth, physiology and nitrogen metabolism were analyzed. Nitrate-grown pioneer species had much higher leaf nitrate reductase activity than non-pioneer ones, but non-pioneer seedlings were also able to use nitrate as a nitrogen source. In addition to this remarkable difference between the groups in the capacity for leaf nitrate assimilation, substantial variations in the nitrogen use strategies were observed within the successional classes. Differently from the other non-pioneers, the canopy species C. estrellensis seemed to assimilate nitrate mainly in the leaves. Morphophysiological analyses showed a gradient of ammonium toxicity response, with E. brasiliensis as the most tolerant species, and T. micrantha and H. popayanensis as the most sensitive ones. Guarea kunthiana showed a relatively low tolerance to ammonium and an unusual high translocation of this cation in the xylem sap. In contrast to the other pioneers, C. pachystachya had a high plasticity in the use of nitrogen sources. Overall, these results suggest that nitrogen use strategies of neotropical tree seedlings were not determined solely by their successional position. Copyright © 2017 Elsevier Masson SAS. All rights reserved.

Leaf litter breakdown rates and associated fauna of native and exotic trees used in Neotropical Riparia Reforestation; Tasas de perdida de masa de la hojarasca y fauna asociada en especies de arboles comunmente utilizados en la Reforestacion de Riberas Neotropicales

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Gutierrez Isaza, Nataly; Blanco, Juan Felipe

2014-07-01

A signature of globalization is the prevalence of exotic trees along reforested urban and rural riparian zones in the neotropics, but little is known about the instream processing of its leaf litter. In this study, leaf litter breakdown rates were measured during 35 days using mesh bags within a reference headwater stream for seven exotic and three native tree species commonly used in urban and rural reforestation. Artocarpus altilis, Schefflera actinophylla and Terminalia catappa scored the highest mass loss rates (>85 %; mean life: t50 <15 d), while Cecropia sp. and Cespedesia macrophylla (mass loss =36 and 15 %; t50 =58 and 172 d, respectively) scored the lowest rates. However, a broad range of rates was observed among the ten species studied. The carbon to phosphorus ratio (c:p) and toughness of the leaf litter were the best predictors of breakdown rates. However, these leaf properties were not correlated with the very low values of macro invertebrates abundance and diversity, and the few morpho classified as shredders. Therefore physical rather than biological controls seem to best explain the observed variability of mass loss rates, and thus slow decomposing leaf litter species seems to provide a habitat rather than a food resource, particularly to collectors. This study suggests that riparian reforestation will propagate species-specific ecological influences on instream processes such as leaf litter processing depending on leaf quality properties, therefore ecosystem-wide influences should be considered for improving reforestation strategies. Future studies should test for differences in breakdown rates and colonization by macro invertebrates relative for leaf litter species origin (native vs. exotic).

Leaf litter breakdown rates and associated fauna of native and exotic trees used in Neotropical Riparia Reforestation

International Nuclear Information System (INIS)

Gutierrez Isaza, Nataly; Blanco, Juan Felipe

2014-01-01

A signature of globalization is the prevalence of exotic trees along reforested urban and rural riparian zones in the neotropics, but little is known about the instream processing of its leaf litter. In this study, leaf litter breakdown rates were measured during 35 days using mesh bags within a reference headwater stream for seven exotic and three native tree species commonly used in urban and rural reforestation. Artocarpus altilis, Schefflera actinophylla and Terminalia catappa scored the highest mass loss rates (>85 %; mean life: t50 <15 d), while Cecropia sp. and Cespedesia macrophylla (mass loss =36 and 15 %; t50 =58 and 172 d, respectively) scored the lowest rates. However, a broad range of rates was observed among the ten species studied. The carbon to phosphorus ratio (c:p) and toughness of the leaf litter were the best predictors of breakdown rates. However, these leaf properties were not correlated with the very low values of macro invertebrates abundance and diversity, and the few morpho classified as shredders. Therefore physical rather than biological controls seem to best explain the observed variability of mass loss rates, and thus slow decomposing leaf litter species seems to provide a habitat rather than a food resource, particularly to collectors. This study suggests that riparian reforestation will propagate species-specific ecological influences on instream processes such as leaf litter processing depending on leaf quality properties, therefore ecosystem-wide influences should be considered for improving reforestation strategies. Future studies should test for differences in breakdown rates and colonization by macro invertebrates relative for leaf litter species origin (native vs. exotic).

Recovery after 25 years of the tree and palms species diversity on a selectively logged forest in a Venezuelan lowland ecosystem

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Lozada, J.R.; Arends, E.; Sánchez, D.; Villarreal, A.; Guevara, J.; Soriano, P.; Costa, M.

2016-07-01

Aim of the study: We evaluate palm and tree species diversity in a floodplain forest and the changes affecting the plots subjected to different intensities of selective logging. Area of study: The western alluvial plains of Venezuela. Materials and Methods: A randomized complete blocks design was established 25 years ago with three felling treatments (trees with diameter greater than 20 cm, 40 cm and 60 cm). Each treatment had three replications, using 1 ha permanent plots. We have measured all trees and palms bigger than over 10 cm in diameter. The data set was used to calculate the Importance Value Index of each species, the Shannon-Wiener index, the Hill Numbers and the Chao-Sørensen index. Main results: Disturbance increases the importance value index of pioneer species like Cecropia peltata, Ochroma pyramidale and Triplaris americana. All treatments produce changes on the floristic diversity but most of them are not significant. Only the high impact treatment causes a decrease in the species richness, but after 5 year of recovery this parameter is close to its previous levels (N0= 43.5). In logged forests, species loss (9.2%) is lower than in the control plots (11.7%) and is also lower than the rate of occurrence of species input (14.6%). Research highlights: In these logged forests restoration of diversity is acceptable because is higher than 91% (Chao-Sørensen index). Selective logging, with low and medium intensity, is a disturbance that works in a similar way to natural disturbances. All the diversity indexes recovered the pre-harvest level values. (Author)

ORIGEN GEOGRÁFICO Y BOTÁNICO DE MIELES DE Apis mellifera (APIDAE EN CUATRO DEPARTAMENTOS DE COLOMBIA

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Guiomar Nates Parra

2013-09-01

Full Text Available El objetivo de este trabajo fue determinar marcadores palinológicos que permitieran caracterizar el origen geográfico y botánico de mieles provenientes de los departamentos de Boyacá, Cundinamarca, Santander y Magdalena. Se realizaron análisis melisopalinológicos de 184 muestras de miel procedentes de 131 apiarios. Se determinaron diferencias significativas entre tipos de mieles mediante un análisis discriminante y comparando la composición de especies entre las muestras. En total se encontraron 297 especies distribuidas en 69 familias, dentro de las cuales las más representativas fueron Mimosa sp., Cecropia sp., Eucalyptus sp., Piper sp. y Quercus humboldtii. Las familias más importantes fueron Fabaceae, Asteraceae,Myrtaceae, Rubiaceae, Fagaceae, y Melastomataceae. Se lograron determinar seis grupos de mieles diferenciadas por su origen geográfico: Altiplano Cundiboyacense, Medio Chicamocha, Sumapaz, Bajo Chicamocha, Sierra Nevada de Santa Marta y Provincia Comunera; también se encontraron diferencias entre las mieles de las regiones andinas y subandinas. Dentro de los tipos de mieles diferenciadas por origen botánico predominaron las mieles monoflorales de Trifolium pratense, Coffea arabica, Eucalyptus sp., Inga sp.y Heliocarpus americanus, mieles oligoflorales de asteráceas y mezclas de mielato de Q. humboldtii y néctar floral (Eucalyptus sp. tipo Brassicaceae, asteráceas. La información de este trabajo junto con la obtenida en análisis fisicoquímicos y sensoriales servirá de base para que los apicultores puedan solicitar la denominación de origen de estas mieles.

Evaluación de la capacidad antioxidante y metabolitos secundarios de extractos de dieciséis plantas medicinales

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Echavarría, Ana

2016-09-01

Full Text Available This study evaluated the antioxidant capacity of sixteen medicinal plants: Escoba amarga (Parthenium hysterophons, ajenjo (Artemisia absinthium, guarumo (Cecropia obtusifolia, chaya (Cnidoscolus chayamansa, borraja (Borago officinalis, balsa (Ochroma sp., linaza (Linum usitatissimum, hierba Luisa (Cymbopogon citratus, toronjil (Melissa officinalis, buganvilla (Bougainvillea spectabilis, alcachofa (Cynara scolymus, guaviduca (Piper carpunya, altamisa (Ambrosia cumanensis, diente de León (Taxacum officinales, buscapina (Parietaria officinalisand moringa (Moringa oleifera. For this, the DPPH (radical 1, 1-difenil-2-picrilhidrazil method was used; furthermore, recognition assays of secondary metabolites were performed, in order to obtain the first signs of phytochemical compounds of interest. The free radical scavenging activity of the extracts was expressed as IC value (g/mL (necessary amount to inhibit the formation of 50% of DPPH radical. The low value of IC50 reflects better free radical scavenging action. Although most of the samples tested showed good antioxidant capacity with this method (DPPH, tests of hydroalcoholic extracts show that alcachofa (IC50 9.89 mg/mL, moringa (IC50 11.4 mg/mL and borraja (IC50 14.0 mg/mL were those with higher antioxidant capacity. Through chemical characterization tests, the presence of flavonoids, tannins, triterpenes, alkaloids and saponins were detected in most of the analyzed species (approximately 56-69%; only 20% of them showed the presence of polyphenols, cyanogenic glycosides, lactones, coumarins, anthraquinones and sterols. According to the results obtained, these plants might be considered as promising sources of secondary metabolites with antioxidant activity.

Serratia marcescens Induces Apoptotic Cell Death in Host Immune Cells via a Lipopolysaccharide- and Flagella-dependent Mechanism*

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Ishii, Kenichi; Adachi, Tatsuo; Imamura, Katsutoshi; Takano, Shinya; Usui, Kimihito; Suzuki, Kazushi; Hamamoto, Hiroshi; Watanabe, Takeshi; Sekimizu, Kazuhisa

2012-01-01

Injection of Serratia marcescens into the blood (hemolymph) of the silkworm, Bombyx mori, induced the activation of c-Jun NH2-terminal kinase (JNK), followed by caspase activation and apoptosis of blood cells (hemocytes). This process impaired the innate immune response in which pathogen cell wall components, such as glucan, stimulate hemocytes, leading to the activation of insect cytokine paralytic peptide. S. marcescens induced apoptotic cell death of silkworm hemocytes and mouse peritoneal macrophages in vitro. We searched for S. marcescens transposon mutants with attenuated ability to induce apoptosis of silkworm hemocytes. Among the genes identified, disruption mutants of wecA (a gene involved in lipopolysaccharide O-antigen synthesis), and flhD and fliR (essential genes in flagella synthesis) showed reduced motility and impaired induction of mouse macrophage cell death. These findings suggest that S. marcescens induces apoptosis of host immune cells via lipopolysaccharide- and flagella-dependent motility, leading to the suppression of host innate immunity. PMID:22859304

A possible role for the immune system in adult neurogenesis: new insights from an invertebrate model.

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Harzsch, Steffen; von Bohlen Und Halbach, Oliver

2016-04-01

Persistent neurogenesis in the adult brain of both vertebrates and invertebrates was previously considered to be driven by self-renewing neuronal stem cells of ectodermal origin. Recent findings in an invertebrate model challenge this view and instead provide evidence for a recruitment of neuronal precursors from a non-neuronal source. In the brain of adult crayfish, a neurogenic niche was identified that contributes progeny to the adult central olfactory pathway. The niche may function in attracting cells from the hemolymph and transforming them into cells with a neuronal fate. This finding implies that the first-generation neuronal precursors located in the crayfish neurogenic niche are not self-renewing. Evidence is summarized in support of a critical re-evaluation of long-term self-renewal of mammalian neuronal stem cells. Latest findings suggest that a tight link between the immune system and the system driving adult neurogenesis may not only exist in the crayfish but also in mammals. Copyright © 2015 Elsevier GmbH. All rights reserved.

Trophic transference of microplastics under a low exposure scenario: Insights on the likelihood of particle cascading along marine food-webs.

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Santana, M F M; Moreira, F T; Turra, A

2017-08-15

Microplastics are emergent pollutants in marine environments, whose risks along food-web still need to be understood. Within this knowledge gap, MPs transference and persistence along trophic levels are key processes. We assessed the potential occurrence of these processes considering a less extreme scenario of exposure than used previously, with microplastics present only in the hemolymph of prey (the mussel Perna perna) and absent in the gut cavity. Predators were the crab Callinectes ornatus and the puffer fish Spheoeroides greeleyi. Transference of microplastics occurred from prey to predators but without evidences of particle persistence in their tissues after 10days of exposure. This suggests a reduced likelihood of trophic cascading of particles and, consequently, a reduced risk of direct impacts of microplastics on higher trophic levels. However, the contact with microplastics along food-webs is still concerning, modulated by the concentration of particles in prey and predators' depuration capacity and rate. Copyright © 2017. Published by Elsevier Ltd.

Detection of Panulirus argus Virus 1 in Caribbean spiny lobsters.

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Montgomery-Fullerton, Megan M; Cooper, Roland A; Kauffman, Kathryn M; Shields, Jeffrey D; Ratzlaff, Robert E

2007-06-07

Panulirus argus Virus 1 (PaV1) is a pathogenic virus that infects Caribbean spiny lobsters P. argus in the Florida Keys. We have developed a PCR detection assay for PaV1 for the purpose of studying the natural history of the virus and for monitoring the prevalence of infection. The detection of the virus in hemolymph and other tissues is based on the PCR amplification of a 499 bp product using specific primers designed from a cloned fragment of the PaV1 genome. The sensitivity limit for the assay was 1.2 fg of purified viral DNA. The PaV1 primers did not react with lobster DNA, oyster DNA, Ostreid Herpesvirus 1, or murine cytomegalovirus. Using this assay, we successfully followed the course of infection in lobsters inoculated with PaV1 and we detected infections in wild-caught lobsters from the Florida Keys. We have also established guidelines for interpreting infection results from the PCR assay for PaV1.

Intracellular survival of Staphylococcus aureus during persistent infection in the insect Tenebrio molitor.

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McGonigle, John E; Purves, Joanne; Rolff, Jens

2016-06-01

Survival of bacteria within host cells and tissues presents a challenge to the immune systems of higher organisms. Escape from phagocytic immune cells compounds this issue, as immune cells become potential vehicles for pathogen dissemination. However, the duration of persistence within phagocytes and its contribution to pathogen load has yet to be determined. We investigate the immunological significance of intracellular persistence within the insect model Tenebrio molitor, assessing the extent, duration and location of bacterial recovery during a persistent infection. Relative abundance of Staphylococcus aureus in both intracellular and extracellular fractions was determined over 21 days, and live S. aureus were successfully recovered from both the hemolymph and within phagocytic immune cells across the entire time course. The proportion of bacteria recovered from within phagocytes also increased over time. Our results show that to accurately estimate pathogen load it is vital to account for bacteria persisting within immune cells. Copyright © 2016 Elsevier Ltd. All rights reserved.

Necrosis-Driven Systemic Immune Response Alters SAM Metabolism through the FOXO-GNMT Axis

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Fumiaki Obata

2014-05-01

Full Text Available Sterile inflammation triggered by endogenous factors is thought to contribute to the pathogenesis of acute and chronic inflammatory diseases. Here, we demonstrate that apoptosis-deficient mutants spontaneously develop a necrosis-driven systemic immune response in Drosophila and provide an in vivo model for studying the organismal response to sterile inflammation. Metabolomic analysis of hemolymph from apoptosis-deficient mutants revealed increased sarcosine and reduced S-adenosyl-methionine (SAM levels due to glycine N-methyltransferase (Gnmt upregulation. We showed that Gnmt was elevated in response to Toll activation induced by the local necrosis of wing epidermal cells. Necrosis-driven inflammatory conditions induced dFoxO hyperactivation, leading to an energy-wasting phenotype. Gnmt was cell-autonomously upregulated by dFoxO in the fat body as a possible rheostat for controlling energy loss, which functioned during fasting as well as inflammatory conditions. We propose that the dFoxO-Gnmt axis is essential for the maintenance of organismal SAM metabolism and energy homeostasis.

Acquisition and transmission of Hepatozoon canis (Apicomplexa: Hepatozoidae) by the tick Amblyomma ovale (Acari: Ixodidae).

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Rubini, A S; Paduan, K S; Martins, T F; Labruna, M B; O'Dwyer, L H

2009-10-14

The present study aimed to evaluate under controlled conditions the acquisition of Hepatozoon canis by Amblyomma ovale after feeding on infected dogs, and the subsequent induction of infection in uninfected dogs that ingested the experimentally infected ticks. Two H. canis naturally infected dogs were infested with A. ovale adult ticks derived from an uninfected laboratory tick colony. After feeding, two A. ovale females presented H. canis oocysts in the hemolymph at the first and fourth days after removal of ticks from dogs. The oocysts had an average size of 244.34 microm x 255.46 microm. Three uninfected dogs were fed with ticks previously fed on the infected dogs. Only one dog became infected 32 days after oral inoculation, presenting circulating gametocytes, parasitemia less than 1%, and positive PCR confirmed to be H. canis by DNA sequencing. The results obtained indicated A. ovale ticks as potential vector of H. canis in rural areas of Brazil.

Crystallization and preliminary X-ray analysis of cecropin B from Bombyx mori

International Nuclear Information System (INIS)

Liu, Zhongyuan; Zhou, Qiangjun; Mao, Xinfang; Zheng, Xiangdong; Guo, Jiubiao; Zhang, Fuchun; Wen, Tingyi; Pang, Hai

2010-01-01

Cecropin B derived from the hemolymph of Bombyx mori has been crystallized by the hanging-drop vapour-diffusion method. The crystal diffracted to 1.43 Å resolution using X-ray radiation. Cecropin B is a 37-residue cationic antimicrobial peptide derived from the haemolymph of Bombyx mori. The precise mechanism by which cecropins exert their antimicrobial and cytolytic activities is not well understood. Crystals of cecropin B were obtained by the hanging-drop vapour-diffusion method using polyethylene glycol as a precipitant at 289 K. The crystal diffracted to 1.43 Å resolution using X-ray radiation and belonged to the orthorhombic space group P1, with unit-cell parameters a = 15.08, b = 22.75, c = 30.20 Å, α = 96.9, β = 103.1, γ = 96.5°. The asymmetric unit contained only one molecule of cecropin B, with a calculated Matthews coefficient of 2.48 Å 3 Da −1 and a solvent content of 50.4%

Crystal structure of Bombyx mori arylphorins reveals a 3:3 heterohexamer with multiple papain cleavage sites

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Hou, Yong; Li, Jianwei; Li, Yi; Dong, Zhaoming; Xia, Qingyou; Yuan, Y Adam

2014-01-01

In holometabolous insects, the accumulation and utilization of storage proteins (SPs), including arylphorins and methionine-rich proteins, are critical for the insect metamorphosis. SPs function as amino acids reserves, which are synthesized in fat body, secreted into the larval hemolymph and taken up by fat body shortly before pupation. However, the detailed molecular mechanisms of digestion and utilization of SPs during development are largely unknown. Here, we report the crystal structure of Bombyx mori arylphorins at 2.8 Å, which displays a heterohexameric structural arrangement formed by trimerization of dimers comprising two structural similar arylphorins. Our limited proteolysis assay and microarray data strongly suggest that papain-like proteases are the major players for B. mori arylphorins digestion in vitro and in vivo. Consistent with the biochemical data, dozens of papain cleavage sites are mapped on the surface of the heterohexameric structure of B. mori arylphorins. Hence, our results provide the insightful information to understand the metamorphosis of holometabolous insects at molecular level. PMID:24639361

A novel immune-related gene HDD1 of silkworm Bombyx mori is involved in bacterial response.

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Zhang, Kui; Pan, Guangzhao; Zhao, Yuzu; Hao, Xiangwei; Li, Chongyang; Shen, Li; Zhang, Rui; Su, Jingjing; Cui, Hongjuan

2017-08-01

Insects have evolved an effective immune system to respond to various challenges. In this study, a novel immune-related gene, called BmHDD1, was first charactered in silkworm, Bombyx mori. BmHDD1 contained an ORF of 837bp and encoding a deduced protein of 278 amino acids. BmHDD1 was specifically expressed in hemocytes, and highly expressed at the molting and metamorphosis stages under normal physiological conditions. Our results suggested that BmHDD1 was mainly generated by hemocytes and secreted into hemolymph. Our results also showed that the expression level of BmHDD1 was significantly increased after 20E injection, which indicated that BmHDD1 might be regulated by ecdysone. More importantly, BmHDD1 was dramatically induced after injected with different types of PAMPs or bacteria, either in hemocytes or fat body. Those results suggested that BmHDD1 plays a role in developing and immunity system in silkworm, Bombyx mori. Copyright © 2017. Published by Elsevier Ltd.

Immunosuppression in Honeybee Queens by the Neonicotinoids Thiacloprid and Clothianidin.

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Brandt, Annely; Grikscheit, Katharina; Siede, Reinhold; Grosse, Robert; Meixner, Marina Doris; Büchler, Ralph

2017-07-05

Queen health is crucial to colony survival of honeybees, since reproduction and colony growth rely solely on the queen. Queen failure is considered a relevant cause of colony losses, yet few data exist concerning effects of environmental stressors on queens. Here we demonstrate for the first time that exposure to field-realistic concentrations of neonicotinoid pesticides can severely affect the immunocompetence of queens of western honeybees (Apis mellifera L.). In young queens exposed to thiacloprid (200 µg/l or 2000 µg/l) or clothianidin (10 µg/l or 50 µg/l), the total hemocyte number and the proportion of active, differentiated hemocytes was significantly reduced. Moreover, functional aspects of the immune defence namely the wound healing/melanisation response, as well as the antimicrobial activity of the hemolymph were impaired. Our results demonstrate that neonicotinoid insecticides can negatively affect the immunocompetence of queens, possibly leading to an impaired disease resistance capacity.

Enzyme Histochemistry for Functional Histology in Invertebrates.

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Cima, Francesca

2017-01-01

In invertebrates, enzyme histochemistry has recently found a renaissance regarding its applications in morphology and ecology. Many enzyme activities are useful for the morphofunctional characterization of cells, as biomarkers of biological and pathologic processes, and as markers of the response to environmental stressors. Here, the adjustments to classic techniques, including the most common enzymes used for digestion, absorption, transport, and oxidation, as well as techniques for azo-coupling, metal salt substitution and oxidative coupling polymerization, are presented in detail for various terrestrial and aquatic invertebrates. This chapter also provides strategies to solve the problems regarding anesthesia, small body size, the presence of an exo- or endoskeleton and the search for the best fixative in relation to the internal fluid osmolarity. These techniques have the aim of obtaining good results for both the pre- and post-embedding labeling of specimens, tissue blocks, sections, and hemolymph smears using both light and transmission electron microscopy.

Effects of azadirachtin on Rhodnius prolixus: immunity and trypanosoma interaction

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Patricia de Azambuja

1992-01-01

Full Text Available The effects of azadirachtin, a tetranortriterpenoid from the neem tree Aradirachta indica J. on both immunity and Trypanosoma cruzi interaction within Rhodniusprolixus and other triatomines, were presented Given through a blood meal, azadirachtin affected the immune reactivity as shown by a significant reduction in numbers of hemocytes and consequently nodule formation follwing challenge with Enterobacter cloacae ß12, reduction in ability to produce antibacterial activities in the hemolymph when injected with bacteria, and decreased ability to destroy the infection caused by inoculation of E. cloacae cells. A single dose of azadirachtin was able to block the development of T. cruzi in R. prolixus if given through the meal at different intervals, together with, before or after parasite infection. Similary, these results were observed with different triatomine species and different strains of T. cruzi. Azadirachtin induced a permanent resistance of the vector against reinfection with T. cruzi. The significance of these data is discussed in relation to the general mode of azadirachtin action in insects.

Physiological markers in insects indicating treatment with ionizing radiation

International Nuclear Information System (INIS)

Nation, J.L.; Smittle, B.J.; Milne, K.R.

1999-01-01

Seven markers or tests that can be applied to 3rd instars of the Caribbean fruit fly as indicators of exposure to ionizing radiation are described, including (1) whole body melanization, (2) phenoloxidase spot test, (3) quantitative phenoloxidase measurement, (4) measurement of the ratio between size of the supraesophageal ganglion and the proventriculus, (5) development of imaginal discs, (6) number of hemocytes in one μl of hemolymph, and (7) larval weight. The markers work best and are most definitive when larvae are exposed to at least 50 Gy no later than the first day of the 3rd instar. All of the tests are sensitive enough to be applied to a single 3rd instar larva. Combinations of some of the tests could be used on a single larva. Tests (1) and (2) are easiest to use and require no specific technical training, and seem to have the most potential for practical use in quarantine. (author)

Evidence for exploitative competition: Comparative foraging behavior and roosting ecology of short-tailed fruit bats (Phyllostomidae)

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Bonaccorso, F.J.; Winkelmann, J.R.; Shin, D.; Agrawal, C.I.; Aslami, N.; Bonney, C.; Hsu, A.; Jekielek, P.E.; Knox, A.K.; Kopach, S.J.; Jennings, T.D.; Lasky, J.R.; Menesale, S.A.; Richards, J.H.; Rutland, J.A.; Sessa, A.K.; Zhaurova, L.; Kunz, T.H.

2007-01-01

Chestnut short-tailed bats, Carollia castanea, and Seba's short-tailed bats, C. perspicillata (Phyllostomidae), were radio-tracked (N = 1593 positions) in lowland rain forest at Tiputini Biodiversity Station, Orellana Province, Ecuador. For 11 C. castanea, mean home range was 6.8 ?? 2.2 ha, mean core-use area was 1.7 ?? 0.8 ha, and mean long axis across home range was 438 ?? 106 m. For three C. perspicillata, mean home range was 5.5 ?? 1.7 ha, mean core-use area was 1.3 ?? 0.6 ha, and mean long axis was 493 ?? 172 m. Groups of less than five C. castanea occupied day-roosts in earthen cavities that undercut banks the Tiputini River. Carollia perspicillata used tree hollows and buildings as day-roosts. Interspecific and intraspecific overlap among short-tailed bats occurred in core-use areas associated with clumps of fruiting Piper hispidum (peppers) and Cecropia sciadophylla. Piper hispidum seeds were present in 80 percent of the fecal samples from C. castanea and 56 percent of samples from C. perspicillata. Carollia perspicillata handled pepper fruits significantly faster than C. castanea; however, C. castanea commenced foraging before C. perspicillata emerged from day-roosts. Evidence for exploitative competition between C. castanea and C. perspicillata is suggested by our observations that 95 percent of ripe P. hispidum fruits available at sunset disappear before sunrise (N = 74 marked fruits). Piper hispidum plants produced zero to 12 ripe infructescences per plant each night during peak production. Few ripe infructescences of P. hispidum were available during the dry season; however, ripe infructescences of C. sciadophylla, remained abundant. ?? 2007 The Author(s) Journal compilation ?? 2007 by The Association for Tropical Biology and Conservation.

Local ecological knowledge and its relationship with biodiversity conservation among two Quilombola groups living in the Atlantic Rainforest, Brazil.

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Conde, Bruno Esteves; Ticktin, Tamara; Fonseca, Amanda Surerus; Macedo, Arthur Ladeira; Orsi, Timothy Ongaro; Chedier, Luciana Moreira; Rodrigues, Eliana; Pimenta, Daniel Sales

2017-01-01

Information on the knowledge, uses, and abundance of natural resources in local communities can provide insight on conservation status and conservation strategies in these locations. The aim of this research was to evaluate the uses, knowledge and conservation status of plants in two Quilombolas (descendants of slaves of African origin) communities in the Atlantic rainforest of Brazil, São Sebastião da Boa Vista (SSBV) and São Bento (SB). We used a combination of ethnobotanical and ecological survey methods to ask: 1) What ethnobotanical knowledge do the communities hold? 2) What native species are most valuable to them? 3) What is the conservation status of the native species used? Thirteen local experts described the names and uses of 212 species in SSBV (105 native species) and 221 in SB (96 native species). Shannon Wiener diversity and Pielou's Equitability indices of ethnobotanical knowledge of species were very high (5.27/0.96 and 5.28/0.96, respectively). Species with the highest cultural significance and use-value indexes in SSBV were Dalbergia hortensis (26/2.14), Eremanthus erythropappus (6.88/1), and Tibouchina granulosa (6.02/1); while Piptadenia gonoacantha (3.32/1), Sparattosperma leucanthum (3.32/1) and Cecropia glaziovii (3.32/0.67) were the highest in SB. Thirty-three native species ranked in the highest conservation priority category at SSBV and 31 at SB. D. hortensis was noteworthy because of its extremely high cultural importance at SSBV, and its categorization as a conservation priority in both communities. This information can be used towards generating sustainable use and conservation plans that are appropriate for the local communities.

Atlantic frugivory: a plant-frugivore interaction data set for the Atlantic Forest.

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Bello, Carolina; Galetti, Mauro; Montan, Denise; Pizo, Marco A; Mariguela, Tatiane C; Culot, Laurence; Bufalo, Felipe; Labecca, Fabio; Pedrosa, Felipe; Constantini, Rafaela; Emer, Carine; Silva, Wesley R; da Silva, Fernanda R; Ovaskainen, Otso; Jordano, Pedro

2017-06-01

The data set provided here includes 8,320 frugivory interactions (records of pairwise interactions between plant and frugivore species) reported for the Atlantic Forest. The data set includes interactions between 331 vertebrate species (232 birds, 90 mammals, 5 fishes, 1 amphibian, and 3 reptiles) and 788 plant species. We also present information on traits directly related to the frugivory process (endozoochory), such as the size of fruits and seeds and the body mass and gape size of frugivores. Data were extracted from 166 published and unpublished sources spanning from 1961 to 2016. While this is probably the most comprehensive data set available for a tropical ecosystem, it is arguably taxonomically and geographically biased. The plant families better represented are Melastomataceae, Myrtaceae, Moraceae, Urticaceae, and Solanaceae. Myrsine coriacea, Alchornea glandulosa, Cecropia pachystachya, and Trema micrantha are the plant species with the most animal dispersers (83, 76, 76, and 74 species, respectively). Among the animal taxa, the highest number of interactions is reported for birds (3,883) followed by mammals (1,315). The woolly spider monkey or muriqui, Brachyteles arachnoides, and Rufous-bellied Thrush, Turdus rufiventris, are the frugivores with the most diverse fruit diets (137 and 121 plants species, respectively). The most important general patterns that we note are that larger seeded plant species (>12 mm) are mainly eaten by terrestrial mammals (rodents, ungulates, primates, and carnivores) and that birds are the main consumers of fruits with a high concentration of lipids. Our data set is geographically biased, with most interactions recorded for the southeast Atlantic Forest. © 2017 by the Ecological Society of America.

ECOLOGICAL FACTORS ASSOCIATED WITH THE ESTABLISHMENT OF RUSTIC SPECIES IN DISTURBED ECOSYSTEMS IN THE ATLANTIC FOREST, PIRAÍ, RIO DE JANEIRO STATE – BRAZIL

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Hiram Feijó Baylão Junior

2013-08-01

Full Text Available http://dx.doi.org/10.5902/1980509810542 Disturbed ecosystems do not present original floristic composition. Their soils are depleted, shallow, stony, with low infiltration and present erosion with different levels of Geodynamics. The region has areas of pasture with sparse herbaceous vegetation which is weakened in every rain and fire, covering less ground. The individuals that colonized and settled in these environments were considered rustic species. This study raised and identified by census, the rustic species and their structure in the most degraded part (lower third of the watershed of Cacaria’s river at the base of the Serra do Mar, Piraí, Rio de Janeiro state, and evaluated the influence of ecological exposure, slope, elevation, topography and rock outcrops in the establishment and growth of these species. For the vegetation survey it was conducted census in an area of 22 hectares, where it was measured, geo-referenced and identified all spontaneous tree species that were isolated in a pasture area. Ecological factors exposure, elevation and slope were determined with a compass, altimeter and clinometer, respectively. We identified 131 individuals, representing 14 species, grouped into nine families. Tabernaemontana laeta Mart., Sparattosperma leucanthum (Vell. Schum., Machaerium hirtum (Vell. Stellfeld, Tabebuia chrysotricha (Mart. ex DC. Stan., Cecropia pachystachya Trec., Peltophorum dubium (Spreng. Taub., Guarea guidonia (L. Sleumer, Acacia polyphylla DC. and Psidium guajava L.were present in portions of the slope with exposure to the north, with altitudes from 60m to 80m and with slope strongly corrugated (20-45%, indicating a preference of these species for microhabitats with those characteristics. 

Floristic composition and soil fertility in gold mining of Lavrinha, Pontes and Lacerda, MT, Brazil

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Tatiani Botini Pires

2011-12-01

Full Text Available The mining activities cause intense environmental degradation, especially to the soil and vegetation. This study evaluated attributes of floristic composition and soil fertility in areas of open gold mining areas. One hundred plots of 10 × 10 m were distributed in the area effectively mined and in its surroundings. Stem circunference measures (AGC of all identified woody individuals with greater than or equal to 9 cm were taken at 0.3 m height above the ground. In all plots soil samples at 0-20 cm depth were collected for analysis of fertility attributes that are routinely evaluated. In total we observed the occurrence of 92 species and 43 families. The most abundant families were Myrtaceae, Fabaceae and Dilleniaceae; and families that had the greatest numbers of species were Fabaceae (10, Malpighiaceae (7 and Vochysiaceae (5. In the area effectively mined the species with higher importance value index (IVI were Curatella americana (89.1 Cecropia hololeuca (40.9, Roupala montana (12.5 and Pouteria ramiflora (10.2 while in surrounding of the mined area the species with the highest IVI were Myrcia multiflora (25.6, Caryocar brasiliense (15.7, Magonia pubescens (14.8 and Vatareia macrocarpa (14.4. In both areas, the soil had low pH, low availability of P, K+, Ca2+ and Mg2+ and high Al3+ saturation. In spite of the low soil fertility of the studied area, it was able to maintain an overall density of vegetation with AGC > 9 cm of 430 individuals per hectare in the effectively mined area and of 2,220 individuals per hectare in their surroundings.

Vegetation changes and human impact inferred from an oxbow lake in southwestern Amazonia, Brazil since the 19th century

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Rodríguez-Zorro, Paula A.; Enters, Dirk; Hermanowski, Barbara; da Costa, Marcondes Lima; Behling, Hermann

2015-10-01

Pollen and X-ray fluorescence spectrometry (XRF) analyses from a 272 cm-long sediment core of Lago Amapá, an oxbow lake in western Amazonia, reveal the first palaeoecological investigation of late Holocene sediments in Acre state, Brazil. Radiocarbon dating of older sediments failed due to re-deposition of organic material but a historical map suggests that lacustrine deposition started at 1900 AD. We detected two periods of changes in sediment and vegetation, dominated by pioneer taxa especially Cecropia. The first period around 1900 AD is documenting an initial oxbow lake, with regular fluvial input (high Ti) and low accumulation of organic matter (low inc/coh ratio). During that period Andean pollen taxa originating from Peruvian Andean headwaters were deposited. A fully lacustrine phase started about 1950 AD and is characterized by prolonged periods of stagnant water (low Fe/Mn ratio). The increase of pioneer taxa, sedimentation rates and a reduction of most of the XRF element counts point to a period during which Lago Amapá was a more isolated lake which was flooded only during exceptional severe flood events and is catching mainly anthropogenic disturbances. The extensive human influence during this period was assumed by 1) the high occurrence of pioneer taxa and the absence of charcoal which could indicate changes in vegetation possibly as a result of logging, 2) the Ca and Ti/K ratio which reflect changes to a local sediment source, and 3) comparison of Landsat images from the last 30 years which shows broad changes in vegetation cover and land transformation in the peripheral areas of the oxbow lake.

INVASIVE WEEDS IN BOGOR BOTANIC GARDENS, INDONESIA AND ITS IMPLICATION ON SURROUNDING LANDSCAPES

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Edi Santosa

2014-07-01

Full Text Available Conservation areas with the objective for collection and exchange plant materials have been speculated as weed bank for surrounding areas. Objective of this study was to identify and characterize ruderal invasive weeds in the Bogor Botanic Gardens (BBG. Observations were conducted in all vak (collection blocks in the BBG in order to identify the weeds species, determine their invasiveness, dominance and distribution. Weeds associations with host plants were observed. Current weed control program and data of dead trees collection were analyzed in relevant to weed. Distribution of weeds outside BBG was observed by transects method following river and road directions. Results showed that there were seven invasive weeds, i.e., Cecropia adenopus (Cecropiaceae, Cissus nodosa Blume (Vitaceae, Cissus sicyoides Blume (Vitaceae, Dioscorea bulbifera L. (Dioscoreaceae, Ficus elastica Roxb. (Moraceae, Mikania micrantha H.B.K. (Asteraceae and Paraserianthes falcataria (L. Nielsen (Fabaceae. These seven weeds species invaded 41 out of 215 plant families in BBG. Six species of weeds, i.e., C. adenopus, C. nodosa Blume., C. sicyoides Blume., D. bulbifera L., M. micrantha H.B.K. and P. falcataria (L. Nielsen, were introduced as BBG collections for the first time while the F. elastica Roxb was considered as native. It is most likely that the weeds dispersal agents are the wind, birds, bats, visitors, and waters. All of these weeds existed in surrounding areas outside BBG. Given the detrimental impact of invasive weeds on the plant collection in BBG, it is necessary to develop long–term comprehensive control measures both inside and neighboring areas by involving other government authorities beyond BBG.

Cardiovascular responses to locomotor activity and feeding in unrestrained three-toed sloths, Bradypus variegatus

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D.P.F. Duarte

2004-10-01

Full Text Available Heart rate (HR and systolic (SBP, diastolic (DBP and mean (MBP blood pressure were recorded by biotelemetry in nine conscious unrestrained sloths for 1 min every 15 min over a 24-h period. The animals were allowed to freely move in an acoustically isolated and temperature-controlled (24 ± 1ºC experimental room with light-dark cycle (12/12 h. Behavior was closely monitored through a unidirectional visor and classified as resting (sitting or suspended, feeding (chewing and swallowing embauba leaves, Cecropia adenops, or locomotor activity around the tree trunk or on the room floor. Locomotor activity caused statistically significant increases in SBP (+8%, from 121 ± 22 to 131 ± 18 mmHg, DBP (+7%, from 86 ± 17 to 92 ± 10 mmHg, MBP (+8%, from 97 ± 19 to 105 ± 12 mmHg, and HR (+14%, from 84 ± 15 to 96 ± 15 bpm compared to resting values, indicating a possible major influence of the autonomic nervous system on the modulation of cardiac function during this behavior. During feeding, the increase in blood pressure was even higher (SBP +27%, from 119 ± 21 to 151 ± 21 mmHg; DBP +21%, from 85 ± 16 to 103 ± 15 mmHg; MBP +24%, from 96 ± 17 to 119 ± 17 mmHg, while HR remained at 14% (from 84 ± 15 to 96 ± 10 bpm above resting values. The proportionally greater increase in blood pressure than in HR during feeding suggests an increase in peripheral vascular resistance as part of the overall response to this behavior.

Laccase 1 gene from Plutella xylostella (PxLac1) and its functions in humoral immune response.

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Wang, Ze-Hua; Hu, Rong-Min; Ye, Xi-Qian; Huang, Jian-Hua; Chen, Xue-Xin; Shi, Min

Laccase (EC 1.10.3.2) is a phenoloxidase found in many insect species. The Laccase 1 gene from Plutella xylostella (PxLac1) was cloned, and its expression patterns and functions were determined using qPCR and RNAi methods. The results showed that the expression levels of PxLac1 were consistently high in all larval stages, and the most abundant was in the midgut during the 4th instar stage. Moreover, the expression of PxLac1 was up-regulated in response to bacterial infection, and decreased 24 h after being parasitized by Cotesia vestalis. Further analyses indicated that the effect of parasitization on PxLac1 was induced by active C. vestalis Bracovirus (CvBV). Haemocyte-free hemolymph phenoloxidase (PO) activity was suppressed when PxLac1 was treated with RNAi. Our results provide evidence for a connection between the Laccase 1 gene and insect immunity, and revealed that parasitoid polydnavirus suppresses host PO activity via PxLac1 regulation. Copyright © 2018. Published by Elsevier Ltd.

Electron microscopy of Drosophila garland cell nephrocytes: Optimal preparation, immunostaining and STEM tomography.

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Hochapfel, Florian; Denk, Lucia; Maaßen, Christine; Zaytseva, Yulia; Rachel, Reinhard; Witzgall, Ralph; Krahn, Michael P

2018-01-29

Due to its structural and molecular similarities to mammalian podocytes, the Drosophila nephrocyte emerged as a model system to study podocyte development and associated diseases. Similar to podocytes, nephrocytes establish a slit diaphragm between foot process-like structures in order to filter the hemolymph. One major obstacle in nephrocyte research is the distinct visualization of this subcellular structure to assess its integrity. Therefore, we developed a specialized dissection and fixation protocol, including high pressure freezing and freeze substitution techniques, to improve the preservation of the intricate ultrastructural details necessary for electron microscopic assessment. By means of scanning transmission electron microscopy (STEM) tomography, a three-dimensional dataset was generated to further understand the complex architecture of the nephrocyte channel system. Moreover, a staining protocol for immunolabeling of ultrathin sections of Epon-embedded nephrocytes is discussed, which allows the reliable detection of GFP-tagged fusion proteins combined with superior sample preservation. Due to the growing number of available GFP-trap fly lines, this approach is widely applicable for high resolution localization studies in wild type and mutant nephrocytes. © 2018 Wiley Periodicals, Inc.

Juvenile hormone-binding proteins of Melanoplus bivittatus identified by EFDA photoaffinity labeling

International Nuclear Information System (INIS)

Winder, B.S.

1988-01-01

Proteins that bind juvenile hormone in the hemolymph and fat body of the grasshopper, Melanoplus bivittatus were identified by photoaffinity labeling with radiolabeled epoxyfarnesyl diazoacetate ( 3 H-EFDA), and were characterized by electrophoretic analysis. A protocol was developed which allowed detection of 3 H-EFDA that was covalently linked to proteins upon exposure to ultraviolet light at 254 nm. Quantification of protein-linked 3 H-EFDA by liquid scintillation spectrometry took advantage of the differential solubility of unlinked 3 H-EFDA in toluene alone, and of the protein-linked 3 H-EFDA in toluene plus the detergent, Triton X-100. Competition between EFDA and juvenile hormone (JH) for binding to JH-specific binding sites was measured by hydroxyapatite protein binding assays in the presence of radiolabeled JH or EFDA and competing non-radiolabeled hormone. The protein-linked EFDA was detected on fluorograms of SDS or nondenaturing polyacrylamide gels (PAGE), and by liquid scintillation spectrometry of membranes to which the proteins had been electrophoretically transferred. Proteins which specifically bound JH were identified by photolabeling proteins in the presence and absence of nonlabeled JH-III

Primeira evidência de Trypanosoma rangeli no sudeste do Brasil, região endêmica para doença de Chagas First evidence of Trypanosoma rangeli in the southeast of Brazil, an endemic region to Chagas' disease

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Luis Eduardo Ramirez

1998-02-01

Full Text Available Informa-se, pela primeira vez, os achados de Trypanosoma rangeli no Triângulo Mineiro, Sudeste do Brasil, área altamente endêmica para doença de Chagas, assim como a infecção natural da espécie Didelphis albiventris.com este mesmo tripanosoma. Estes foram demonstrados por esfregaços sangüíneos, xenodiagnóstico, hemocultura, microhematócrito e PCR. A PCR foi realizada nas fezes e hemolinfa de Triatoma infestans, usando como controle cepas de T. rangeli provenientes da Colômbia.This short communication informs the discovery of Trypanosoma rangeli for the first time at Triângulo Mineiro region, South-east of Brazil, a highly endemic area of Chagas'disease and also the natural infection of Didelphis albiventris with the same trypanosome. Both the findings were demonstrated through blood smears, xenodiagnosis, microhematocrit technics and PCR. The last one was realized in faeces and hemolymph of Triatoma infestans utilizing as controls strains of T. rangeli from Colombia.

Purification and characterization of tenecin 4, a new anti-Gram-negative bacterial peptide, from the beetle Tenebrio molitor.

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Chae, Jun-Ho; Kurokawa, Kenji; So, Young-In; Hwang, Hyun Ok; Kim, Min-Su; Park, Ji-Won; Jo, Yong-Hun; Lee, Yong Seok; Lee, Bok Luel

2012-03-01

The biochemical characterization of novel antimicrobial peptides (AMPs) and the determination of ligand molecules that induce AMP production are essential for understanding the host innate immune response in insects. Here, we purified a new 14-kDa AMP, named tenecin 4, from the larval hemolymph of the beetle Tenebrio molitor. Tenecin 4 contains 14% glycine residues and has moderate similarities both to the C-terminal region of Drosophila attacin and to silk-moth gloverin proteins. Purified tenecin 4 showed bactericidal activity against Gram-negative Escherichia coli but not against Gram-positive Bacillus subtilis or the fungus Candida albicans. Tenecin 4 production was induced by Toll cascade-activating ligands, such as β-1,3-glucan, lysine-type peptidoglycan and active Spätzle, and by the probable Imd pathway-activating ligand monomeric meso-diaminopimelic acid-type peptidoglycan. Taken together, these data show that tenecin 4 is a defense protein against Gram-negative pathogens and is induced by multiple ligands in Tenebrio larvae. Copyright © 2011 Elsevier Ltd. All rights reserved.

Origins and activation of prophenoloxidases in the digestive tract of the cricket, Gryllus bimaculatus.

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Joseph, Woodring

2014-10-01

The function of Phenoloxidases (POs) in sclerotization and defense in insects is well understood, but little is known concerning their occurrence, origins, and function in the digestive tract. In Gyrllus bimaculatus gut all of the PO activity is found in the lumen of the digestive tract, and no detectible activity is found in homogenates of the gut epithelium or secretions from incubated epithelial tissues. Prophenoloxidases (PPOs) are synthesized in the hemocytes of  Bombyx mori and are transported into the cuticle. It is suggested that the PPOs in the caecal lumen of G. bimaculatus likewise are synthesized in hemocytes and are transported by unknown means into the caecal lumen, where they are activated to POs by trypsin. Peristalsis transports the POs both forward into the crop and posterior within the peritrophic membrane into the hind gut. The PPOs in the hemolymph consist of a trimer (270-280 kDa) and a tetramer (340-370 kDa). The active POs in the gut lumen consist of a monomer (85-95 kDa) in addition to an activated trimer and tetramer. © 2014 Wiley Periodicals, Inc.

The Drosophila blood-brain barrier: Development and function of a glial endothelium

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Stefanie eLimmer

2014-11-01

Full Text Available The efficacy of neuronal function requires a well-balanced extracellular ion homeostasis and a steady supply with nutrients and metabolites. Therefore, all organisms equipped with a complex nervous system developed a so-called blood-brain barrier, protecting it from an uncontrolled entry of solutes, metabolites or pathogens. In higher vertebrates, this diffusion barrier is established by polarized endothelial cells that form extensive tight junctions, whereas in lower vertebrates and invertebrates the blood-brain barrier is exclusively formed by glial cells. Here, we review the development and function of the glial blood-brain barrier of Drosophila melanogaster. In the Drosophila nervous system, at least seven morphologically distinct glial cell classes can be distinguished. Two of these glial classes form the blood-brain barrier. Perineurial glial cells participate in nutrient uptake and establish a first diffusion barrier. The subperineurial glial cells form septate junctions, which block paracellular diffusion and thus seal the nervous system from the hemolymph. We summarize the molecular basis of septate junction formation and address the different transport systems expressed by the blood-brain barrier forming glial cells.

Dung beetles eat acorns to increase their ovarian development and thermal tolerance.

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José R Verdú

Full Text Available Animals eat different foods in proportions that yield a more favorable balance of nutrients. Despite known examples of these behaviors across different taxa, their ecological and physiological benefits remain unclear. We identified a surprising dietary shift that confers ecophysiological advantages in a dung beetle species. Thorectes lusitanicus, a Mediterranean ecosystem species adapted to eat semi-dry and dry dung (dung-fiber consumers is also actively attracted to oak acorns, consuming and burying them. Acorn consumption appears to confer potential advantages over beetles that do not eat acorns: acorn-fed beetles showed important improvements in the fat body mass, hemolymph composition, and ovary development. During the reproductive period (October-December beetles incorporating acorns into their diets should have greatly improved resistance to low-temperature conditions and improved ovarian development. In addition to enhancing the understanding of the relevance of dietary plasticity to the evolutionary biology of dung beetles, these results open the way to a more general understanding of the ecophysiological implications of differential dietary selection on the ecology and biogeography of these insects.

Studies of two naturally occurring compounds which effect release of acetylcholine from synaptosomes

International Nuclear Information System (INIS)

Koenig, M.L.

1985-01-01

Two naturally occurring compounds which effect the release of neurotransmitter from synaptosomes have been purified to apparent homogeneity. Iotrochotin (IOT) isolated from wound exudate of the Caribbean purple bleeder sponge promotes release in a manner that is independent of the extracellular Ca 2+ ion concentration. Leptinotarsin (LPT-d), a protein taken from hemolymph of the Colorado potato beetle, Leptinotarsa decemlineata, stimulates Ca 2+ -dependent release. IOT is slightly acidic and has a molecular weight of approximately 18 kD. [ 3 H]acetylcholine which has been introduced into synaptosomes as [ 3 H]choline can be released by IOT. The toxin releasable pool of labelled neurotransmitter is not depleted by depolarization of the synaptosomes with high potassium, and therefore seems to be primarily extravesicular. LPT-d is a larger protein (molecular weight = 45 kD) than IOT, and seems to effect primarily vesicular release by opening at least one type of presynaptic Ca 2+ channel. The facilitatory effects of the toxin on synaptosomal release can be inhibited by inorganic Ca 2+ channel antagonists, but are not generally affected by organic antagonists

Analysis of odorant-binding protein gene family members in the polyembryonic wasp, Copidosoma floridanum: evidence for caste bias and host interaction.

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Donnell, David M

2014-01-01

The polyembryonic wasp Copidosoma floridanum produces two larval castes, soldiers and reproductives, during development within its caterpillar host. Primary structures were determined for 6 odorant-binding protein (OBP) gene family members in Copidosoma and then analyzed alongside two formerly sequenced OBP genes from this wasp. The genes were examined for caste-bias in expression patterns using reverse transcription-polymerase chain reaction (RT-PCR) and in situ expression studies. Six of the 8 genes show a clear bias in gene expression towards one or the other larval caste. Of the 3 distinct in situ probe hybridization patterns observed in this study, none lie in tissues with clear chemosensory functions. Two of the patterns suggest the majority of the Copidosoma OBP gene family members discovered thus far come into contact with host hemolymph. Most of these OBPs are expressed exclusively in the serosal membrane encompassing each of the reproductive larvae. The absence of expression in the membrane surrounding soldier larvae strongly suggests these OBPs are performing caste-specific functions in the host. Copyright © 2013 Elsevier Ltd. All rights reserved.

Osmoregulated Chloride Currents in Hemocytes from Mytilus galloprovincialis.

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Monica Bregante

Full Text Available We investigated the biophysical properties of the transport mediated by ion channels in hemocytes from the hemolymph of the bivalve Mytilus galloprovincialis. Besides other transporters, mytilus hemocytes possess a specialized channel sensitive to the osmotic pressure with functional properties similar to those of other transport proteins present in vertebrates. As chloride fluxes may play an important role in the regulation of cell volume in case of modifications of the ionic composition of the external medium, we focused our attention on an inwardly-rectifying voltage-dependent, chloride-selective channel activated by negative membrane potentials and potentiated by the low osmolality of the external solution. The chloride channel was slightly inhibited by micromolar concentrations of zinc chloride in the bath solution, while the antifouling agent zinc pyrithione did not affect the channel conductance at all. This is the first direct electrophysiological characterization of a functional ion channel in ancestral immunocytes of mytilus, which may bring a contribution to the understanding of the response of bivalves to salt and contaminant stresses.

Ultrastructural and functional characterization of circulating hemocytes from Galleria mellonella larva: Cell types and their role in the innate immunity.

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Wu, Gongqing; Liu, Yi; Ding, Ying; Yi, Yunhong

2016-08-01

Galleria mellonella larvae have been widely used as a model to study the virulence of various human pathogens. Hemocytes play important roles in the innate immune response of G. mellonella. In this study, the hemocytes of G. mellonella larvae were analyzed by transmission electron microscope, light microscope, and cytochemistry. The cytological and morphological analyses revealed four types of hemocytes; Plasmatocytes, granular cells, spherule cells and oenocytoids. Differential hemocyte counts showed that under our conditions plasmatocytes and granular cells were the most abundant circulating cell types in the hemolymph. We also investigated the role of different types of hemocytes in the cellular and humoral immune defenses. The in-vivo experiment showed that plasmatocytes, granular cells and oenocytoids phagocytized FITC-labelled Escherichia coli bacteria in larvae of G. mellonella, whereas the granular cells exhibited the strongest phagocytic ability against these microbial cells. After incubation with L-DOPA, plasmatocytes, granular cells and oenocytoids are stained brown, indicating the presence of phenoloxidase activity. These results shed new light on our understanding of the immune function of G. mellonella hemocytes. Copyright © 2016 Elsevier Ltd. All rights reserved.

Infection of army ant pupae by two new parasitoid mites (Mesostigmata: Uropodina

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Adrian Brückner

2017-10-01

Full Text Available A great variety of parasites and parasitoids exploit ant societies. Among them are the Mesostigmata mites, a particularly common and diverse group of ant-associated arthropods. While parasitism is ubiquitous in Mesostigmata, parasitoidism has only been described in the genus Macrodinychus. Yet information about the basic biology of most Macrodinychus species is lacking. Out of 24 formally described species, information about basic life-history traits is only available for three species. Here we formally describe two new Macrodinychus species, i.e. Macrodinychus hilpertae and Macrodinychus derbyensis. In both species, immature stages developed as ecto-parasitoids on ant pupae of the South-East Asian army ant Leptogenys distinguenda. By piercing the developing ant with their chelicera, the mites apparently suck ant hemolymph, ultimately killing host individuals. We compare infection rates among all studied Macrodinychus species and discuss possible host countermeasures against parasitoidism. The cryptic lifestyle of living inside ant nests has certainly hampered the scientific discovery of Macrodinychus mites and we expect that many more macrodinychid species await scientific discovery and description.

Combined Effect of Cameo2 and CBP on the Cellular Uptake of Lutein in the Silkworm, Bombyx mori

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Dong, Xiao-Long; Chai, Chun-Li; Pan, Cai-Xia; Tang, Hui; Chen, Yan-Hong; Dai, Fang-Yin; Pan, Min-Hui; Lu, Cheng

2014-01-01

Formation of yellow-red color cocoons in the silkworm, Bombyx mori, occurs as the result of the selective delivery of carotenoids from the midgut to the silk gland via the hemolymph. This process of pigment transport is thought to be mediated by specific cellular carotenoids carrier proteins. Previous studies indicated that two proteins, Cameo2 and CBP, are associated with the selective transport of lutein from the midgut into the silk gland in Bombyx mori. However, the exact roles of Cameo2 and CBP during the uptake and transport of carotenoids are still unknown. In this study, we investigated the respective contributions of these two proteins to lutein and β-carotene transport in Bombyx mori as well as commercial cell-line. We found that tissues, expressed both Cameo2 and CBP, accumulate lutein. Cells, co-expressed Cameo2 and CBP, absorb 2 fold more lutein (PBombyx mori. Cameo2 and CBP, as the membrane protein and the cytosol protein, respectively, have the combined effect to facilitate the cellular uptake of lutein. PMID:24475153

The Drosophila blood-brain barrier: development and function of a glial endothelium.

Science.gov (United States)

Limmer, Stefanie; Weiler, Astrid; Volkenhoff, Anne; Babatz, Felix; Klämbt, Christian

2014-01-01

The efficacy of neuronal function requires a well-balanced extracellular ion homeostasis and a steady supply with nutrients and metabolites. Therefore, all organisms equipped with a complex nervous system developed a so-called blood-brain barrier, protecting it from an uncontrolled entry of solutes, metabolites or pathogens. In higher vertebrates, this diffusion barrier is established by polarized endothelial cells that form extensive tight junctions, whereas in lower vertebrates and invertebrates the blood-brain barrier is exclusively formed by glial cells. Here, we review the development and function of the glial blood-brain barrier of Drosophila melanogaster. In the Drosophila nervous system, at least seven morphologically distinct glial cell classes can be distinguished. Two of these glial classes form the blood-brain barrier. Perineurial glial cells participate in nutrient uptake and establish a first diffusion barrier. The subperineurial glial (SPG) cells form septate junctions, which block paracellular diffusion and thus seal the nervous system from the hemolymph. We summarize the molecular basis of septate junction formation and address the different transport systems expressed by the blood-brain barrier forming glial cells.

Infection of army ant pupae by two new parasitoid mites (Mesostigmata: Uropodina).

Science.gov (United States)

Brückner, Adrian; Klompen, Hans; Bruce, Andrew Iain; Hashim, Rosli; von Beeren, Christoph

2017-01-01

A great variety of parasites and parasitoids exploit ant societies. Among them are the Mesostigmata mites, a particularly common and diverse group of ant-associated arthropods. While parasitism is ubiquitous in Mesostigmata, parasitoidism has only been described in the genus Macrodinychus . Yet information about the basic biology of most Macrodinychus species is lacking. Out of 24 formally described species, information about basic life-history traits is only available for three species. Here we formally describe two new Macrodinychus species, i.e. Macrodinychus hilpertae and Macrodinychus derbyensis . In both species, immature stages developed as ecto-parasitoids on ant pupae of the South-East Asian army ant Leptogenys distinguenda . By piercing the developing ant with their chelicera, the mites apparently suck ant hemolymph, ultimately killing host individuals. We compare infection rates among all studied Macrodinychus species and discuss possible host countermeasures against parasitoidism. The cryptic lifestyle of living inside ant nests has certainly hampered the scientific discovery of Macrodinychus mites and we expect that many more macrodinychid species await scientific discovery and description.

Functional characterization of bursicon receptor and genome-wide analysis for identification of genes affected by bursicon receptor RNAi

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Bai, Hua; Palli, Subba R.

2010-01-01

Bursicon is an insect neuropeptide hormone that is secreted from the central nervous system into the hemolymph and initiates cuticle tanning. The receptor for bursicon is encoded by the rickets (rk) gene and belongs to the G protein-coupled receptor (GPCR) superfamily. The bursicon and its receptor regulate cuticle tanning as well as wing expansion after adult eclosion. However, the molecular action of bursicon signaling remains unclear. We utilized RNA interference (RNAi) and microarray to study the function of the bursicon receptor (Tcrk) in the model insect, Tribolium castaneum. The data included here showed that in addition to cuticle tanning and wing expansion reported previously, Tcrk is also required for development and expansion of integumentary structures and adult eclosion. Using custom microarrays, we identified 24 genes that are differentially expressed between Tcrk RNAi and control insects. Knockdown in the expression of one of these genes, TC004091, resulted in the arrest of adult eclosion. Identification of genes that are involved in bursicon receptor mediated biological processes will provide tools for future studies on mechanisms of bursicon action. PMID:20457145

Novel chimeric foot-and-mouth disease virus-like particles harboring serotype O VP1 protect guinea pigs against challenge.

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Li, Haitao; Li, Zhiyong; Xie, Yinli; Qin, Xiaodong; Qi, Xingcai; Sun, Peng; Bai, Xingwen; Ma, Youji; Zhang, Zhidong

2016-02-01

Foot-and-mouth disease is a highly contagious, acute viral disease of cloven-hoofed animal species causing severe economic losses worldwide. Among the seven serotypes of foot-and-mouth disease virus (FMDV), serotype O is predominant, but its viral capsid is more acid sensitive than other serotypes, making it more difficult to produce empty serotype O VLPs in the low pH insect hemolymph. Therefore, a novel chimeric virus-like particle (VLP)-based candidate vaccine for serotype O FMDV was developed and characterized in the present study. The chimeric VLPs were composed of antigenic VP1 from serotype O and segments of viral capsid proteins from serotype Asia1. These VLPs elicited significantly higher FMDV-specific antibody levels in immunized mice than did the inactivated vaccine. Furthermore, the chimeric VLPs protected guinea pigs from FMDV challenge with an efficacy similar to that of the inactivated vaccine. These results suggest that chimeric VLPs have the potential for use in vaccines against serotype O FMDV infection. Copyright © 2015 Elsevier B.V. All rights reserved.

Seasonal inhalant insect allergy: Harmonia axyridis ladybug.

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Goetz, David W

2009-08-01

The exotic Asian lady beetle, Harmonia axyridis, has become a prominent cause of seasonal inhalant allergy (allergic rhinitis, asthma, and urticaria) in the last two decades in North America and Europe after being introduced into the environment as an agricultural pest-control predator. Seeking winter hibernation sites, ladybug swarms will invade human habitats in the fall. Large fall swarms and smaller spring dispersions produce corresponding peaks in ladybug allergy. Ladybug allergy prevalence in endemic areas has been reported as high as 10%. For some individuals ladybug allergy is their first expression of allergic disease. Exposures at home, work, school, and in other settings may be sensitizing. Ladybug hemolymph is the primary source of allergens. Har a 1 and Har a 2 major ladybug allergens have been characterized. 'Reflex bleeding' from tibiofemoral joints (for communication and during alarm) disperses these allergens. Ladybug skin testing should be routine in endemic areas. Avoidance continues to be the first step in treatment. Allergen vaccine therapy may be effective, but a commercial extract of H. axyridis is needed.

Lipoproteins in Drosophila melanogaster—Assembly, Function, and Influence on Tissue Lipid Composition

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Palm, Wilhelm; Sampaio, Julio L.; Brankatschk, Marko; Carvalho, Maria; Mahmoud, Ali; Shevchenko, Andrej; Eaton, Suzanne

2012-01-01

Interorgan lipid transport occurs via lipoproteins, and altered lipoprotein levels correlate with metabolic disease. However, precisely how lipoproteins affect tissue lipid composition has not been comprehensively analyzed. Here, we identify the major lipoproteins of Drosophila melanogaster and use genetics and mass spectrometry to study their assembly, interorgan trafficking, and influence on tissue lipids. The apoB-family lipoprotein Lipophorin (Lpp) is the major hemolymph lipid carrier. It is produced as a phospholipid-rich particle by the fat body, and its secretion requires Microsomal Triglyceride Transfer Protein (MTP). Lpp acquires sterols and most diacylglycerol (DAG) at the gut via Lipid Transfer Particle (LTP), another fat body-derived apoB-family lipoprotein. The gut, like the fat body, is a lipogenic organ, incorporating both de novo–synthesized and dietary fatty acids into DAG for export. We identify distinct requirements for LTP and Lpp-dependent lipid mobilization in contributing to the neutral and polar lipid composition of the brain and wing imaginal disc. These studies define major routes of interorgan lipid transport in Drosophila and uncover surprising tissue-specific differences in lipoprotein lipid utilization. PMID:22844248

A Plant Bacterial Pathogen Manipulates Its Insect Vector's Energy Metabolism

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Hijaz, Faraj; Ebert, Timothy A.; Rogers, Michael E.

2016-01-01

and biochemical levels. The growth of the plant-pathogenic bacteria in the hemolymph of their vectors indicated that the hemolymph contains all the necessary nutrients for their growth. In addition to nutrients, “Candidatus Liberibacter asiaticus” (CLas) can take up energetic nucleotides, such as ATP, from its vector, Diaphorina citri, using ATP translocase. In this study, we found that the CLas pathogen manipulates the energy metabolism of its insect vector. The accumulation of ATP in CLas-infected D. citri psyllids indicated that CLas induces ATP production to fulfill its need for this energetic compound. As a result of ATP accumulation, a shorter life span and altered feeding behavior were observed. These findings increase our knowledge of insect transmission of the persistent-circulative-propagative type of plant pathogens vectored by insects. PMID:28039132

Aspectos estructurales y tipos de vegetación de la isla Mocagua, río Amazonas

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Prieto Adriana

1995-06-01

Full Text Available An analysis of some parameters of vegetation structure (coberture -%-, density and specific diversity is presented for plant formations in the Mocagua island (Colombian Amazonia. The structure of the vegetation types ranging from simple river beach zone, aquatic and floodplain environments, to complex arrangement in forest formations. The average number of individuals with DAP > 10 cm (58/0.1 ha is lower than other similar regions. In addition, we describe the types of vegetation founded according with floristic affinities, wich is related to the water content in the sustrata.Tha natural vegetation comprise from aquatic communities of Oxyearyum eubense - Pontederia rotundifolia, to continental ones of Rheedia madruno- Palieourea eroeea, Triehilia pallida - Sommera sabiceoides, phases of Montriehardia arboreseens and Ceeropia latíloba, Paspalum repens and Ceeropia latíloba, and Leptochloa seabra and Torulinium maeroeephalum.Se presentan los resultados de los análisis sobre parámetros estructurales de la vegetación (cobertura-%-, densidad y diversidad específica de las formaciones vegetales de la isla Mocagua (Amazonas, Colombia. La estructura de los tipos de vegetación diferenciados cubre desde los arreglos simples como la de las zonas de playa, ambientes lacustres e inundables, hasta arreglos complejos con diferentes estratos en formaciones boscosas. El número de individuos con DAP > 10 cm (prom.=58/0.1 ha en comparación con los valores en otras zonas es bajo. Los tipos de vegetación descritos obedecen en su distribución esencialmente al contenido de agua en el sustrato; la vegetación natural comprende desde comunidades acuáticas de Oxycaryum cubense - Pontederia rotundifolia, hasta las continentales de Rheedia. madruno - Palieourea eroeea, Triehilia pallida - Sommera sabieeoides y las fases de Montriehardia arboreseens y Ceeropia latiloba, Paspalum repens y Cecropia latiloba, y la de Leptochloa scabra y Torulinium maerocephalum.

Plantas hipoglicemiantes utilizadas por comunidades tradicionais na Bacia do Alto Paraguai e Vale do Guaporé, Mato Grosso - Brasil

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M. Macedo

Full Text Available No Brasil, com inúmeras espécies tropicais, evidencia-se o conhecimento e uso da vegetação medicinal pelas comunidades quilombolas, ribeirinhas, rurais, tradicionais e indígenas, que são praticadas até os dias atuais. O Estado de Mato Grosso pela sua localização no Planalto Central brasileiro possui uma variedade destas comunidades, que utilizam estas plantas medicinais, levando em consideração o conhecimento popular passado de geração em geração. Esta pesquisa visa identificar a forma de coleta e os usos desse grupo vegetal indicados pelos usuários. O estudo foi realizado em 15 comunidades tradicionais (não indígenas na Bacia do Alto Paraguai e 2 (duas no Vale do Guaporé. A coleta de dados deu-se através de líderes comunitários, benzedeiras, parteiras e usuários utilizando o método qualitativo, mediante abordagem qualitativa, com auxílio de entrevistas estruturadas, semi-estruturadas e abertas. Dentre as espécies identificadas 17 estacam-se para o tratamento do Diabetes, dentre elas citam-se: Anacadium humile; Bauhinia glabra; Cecropia pachystachya; Hancornia speciosa; Heteropteris aphrodisiaca; Leonotis nepetifolia; Momordica charantia; Solanum lycocarpum. Registra-se ainda que as partes mais utilizadas desses vegetais são: folhas, casca do caule, raiz, planta toda, brotos, óleos dos frutos e polvilho dos frutos. De acordo com os usuários a maioria das plantas não podem ser colhidas após o nascer do sol e o preparo deve ser feito com folhas secas ou que tenham sido submetidas ao processo de secagem sobre o fogão de lenha ou em local abafado.

Plant-based medicines for anxiety disorders, Part 1: a review of preclinical studies.

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Sarris, Jerome; McIntyre, Erica; Camfield, David A

2013-03-01

Research in the area of herbal psychopharmacology has revealed a variety of promising medicines that may provide benefit in the treatment of general anxiety and specific anxiety disorders. However, a comprehensive review of plant-based anxiolytics has been absent to date. This article (part 1) reviews herbal medicines for which only preclinical investigations for anxiolytic activity have been performed. In part 2, we review herbal medicines for which there have been clinical investigations for anxiolytic activity. An open-ended, language-restricted (English) search of MEDLINE (PubMed), CINAHL, Scopus and the Cochrane Library databases was conducted (up to 28 October 2012) using specific search criteria to identify herbal medicines that have been investigated for anxiolytic activity. This search of the literature revealed 1,525 papers, from which 53 herbal medicines were included in the full review (having at least one study using the whole plant extract). Of these plants, 21 had human clinical trial evidence (reviewed in part 2), with another 32 having solely preclinical studies (reviewed here in part 1). Preclinical evidence of anxiolytic activity (without human clinical trials) was found for Albizia julibrissin, Sonchus oleraceus, Uncaria rhynchophylla, Stachys lavandulifolia, Cecropia glazioui, Magnolia spp., Eschscholzia californica, Erythrina spp., Annona spp., Rubus brasiliensis, Apocynum venetum, Nauclea latifolia, Equisetum arvense, Tilia spp., Securidaca longepedunculata, Achillea millefolium, Leea indica, Juncus effusus, Coriandrum sativum, Eurycoma longifolia, Turnera diffusa, Euphorbia hirta, Justicia spp., Crocus sativus, Aloysia polystachya, Albies pindrow, Casimiroa edulis, Davilla rugosa, Gastrodia elata, Sphaerathus indicus, Zizyphus jujuba and Panax ginseng. Common mechanisms of action for the majority of botanicals reviewed primarily involve GABA, either via direct receptor binding or ionic channel or cell membrane modulation; GABA transaminase

Estrutura da comunidade de morcegos (Mammalia, Chiroptera da Estação Experimental de Itirapina, estado de São Paulo, Brasil

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Therys M. Sato

2015-01-01

Full Text Available A estrutura de comunidades de morcegos no Brasil ainda é pouco estudada e vêm sofrendo sérias modificações devido à perda de habitats. O principal objetivo do presente trabalho foi estudar a comunidade de morcegos da Estação Experimental de Itirapina, município de Itirapina, Estado de São Paulo. Nesta Estação, além das espécies cultivadas de Eucalyptus e Pinus, há diversas plantas que podem fornecer alimento aos morcegos. Entre julho de 2001 e julho de 2006, foram realizadas 58 sessões noturnas de captura de morcegos com 4 a 12 redes-de-neblina dispostas a cada sessão. Nesse período, foram capturados 720 indivíduos de 16 espécies de morcegos dos quais 13 pertencem à família Phyllostomidae, duas à Vespertilionidae e uma à Molossidae. A curva cumulativa de espécies atingiu o equilíbrio, no qual apenas espécies raras são acrescentadas. Cinco espécies (Artibeus lituratus, Platyrrhinus lineatus, Carollia perspicillata, Glossophaga soricina e Sturnira lilium representaram 80% dos morcegos capturados na Estação Experimental, onde se alimentavam de frutos de plantas pioneiras, tais como Cecropia pachystachya, Solanum spp. e Piper spp. A prevalência do vírus rábico foi zero na amostragem das 10 espécies analisadas. Apesar de estar muito modificada, a Estação Experimental de Itirapina pode ser uma importante área de abrigo e alimento para os morcegos e tem potencial de atuar como corredor entre áreas de Cerrado e Mata Atlântica.

Abundance and frugivory of the toco toucan (Ramphastos toco in a gallery forest in Brazil's Southern Pantanal

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J. Ragusa-Netto

Full Text Available Unlike other toucan species, the Toco toucan (Ramphastos toco - the largest Ramphastidae - usually inhabits dry semi-open areas. This conspicuous canopy frugivore uses a large home range that includes a variety of vegetation types, among which gallery forests are widely cited as important to this species. However, the factors relating to the occurrence of Toco toucans in such habitats are unclear. I studied the abundance of Toco toucans as well as the availability of fleshy fruit in a gallery forest in the southern Pantanal (sub-region of Miranda, Brazil, in order to assess the relationship between these parameters. Also, I examined toucan foraging activity to analyze its relationship with both toucan abundance and fruit availability. The presence of the Toco toucan was more common in the gallery forest from the middle to the end of the dry season and during the middle of the wet season. Toucans foraged for fleshy fruits, mainly Genipa americana, Ficus luschnatiana, and Cecropia pachystachya fruits, feeding mostly on G. americana (by far the favorite food resource and F. luschnatiana fruits during the dry season, while C. pachystachya fruits were important in the wet season. Toco toucans foraged particularly heavily (> 80% of foraging activity on G. americana fruits during the latter part of the dry season, when fleshy fruit availability declined sharply. Toco toucan abundance in the gallery forest was associated with the availability of the most commonly consumed fleshy fruits, and also with its foraging activity. This finding suggests that the Toco toucan moved to the gallery forest periodically in response to the availability of abundant food resources, especially the G. americana fruits widely available and exploited during the severely dry season. Therefore, these fruits potentially contribute to Toco toucan persistence in the South Pantanal during the harshest period of the year.

Abundance and frugivory of the Toco toucan (Ramphastos toco) in a gallery forest in Brazil's southern Pantanal.

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Ragusa-Netto, J

2006-02-01

Unlike other toucan species, the Toco toucan (Ramphastos toco)--the largest Ramphastidae--usually inhabits dry semi-open areas. This conspicuous canopy frugivore uses a large home range that includes a variety of vegetation types, among which gallery forests are widely cited as important to this species. However, the factors relating to the occurrence of Toco toucans in such habitats are unclear. I studied the abundance of Toco toucans as well as the availability of fleshy fruit in a gallery forest in the southern Pantanal (sub-region of Miranda, Brazil), in order to assess the relationship between these parameters. Also, I examined toucan foraging activity to analyze its relationship with both toucan abundance and fruit availability. The presence of the Toco toucan was more common in the gallery forest from the middle to the end of the dry season and during the middle of the wet season. Toucans foraged for fleshy fruits, mainly Genipa americana, Ficus luschnatiana, and Cecropia pachystachya fruits, feeding mostly on G. americana (by far the favorite food resource) and F. luschnatiana fruits during the dry season, while C. pachystachya fruits were important in the wet season. Toco toucans foraged particularly heavily (> 80% of foraging activity) on G. americana fruits during the latter part of the dry season, when fleshy fruit availability declined sharply. Toco toucan abundance in the gallery forest was associated with the availability of the most commonly consumed fleshy fruits, and also with its foraging activity. This finding suggests that the Toco toucan moved to the gallery forest periodically in response to the availability of abundant food resources, especially the G. americana fruits widely available and exploited during the severely dry season. Therefore, these fruits potentially contribute to Toco toucan persistence in the South Pantanal during the harshest period of the year.

Effects of insect and decapod exclusion and leaf litter species identity on breakdown rates in a tropical headwater stream

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José Rincón

2014-04-01

Full Text Available La gran variedad de especies vegetales junto a los ríos tropicales afecta la diversidad de detritos orgánicos que llegan a los arroyos, creando variaciones temporales en cantidad y calidad de hojarasca. Examinamos la influencia de las especies vegetales y de la exclusión de macroinvertebrados sobre la descomposición de hojarasca en la parte alta de un arroyo en las montañas Luquillo de Puerto Rico. Incubamos hojarasca de Dacryodes excelsa (Burseraceae, Guarea guidonia (Meliaceae, Cecropia scheberiana (Moraceae, Manilkara bidentata (Sapotaceae y Prestoea acuminata (Palmae en bolsas descomposición que colocamos en una poza de Quebrada Prieta. Usamos bolsas de malla fina para excluir los macroinvertebrados y de malla gruesa para permitir la entrada a crustáceos decápodos (camarones y cangrejos jóvenes e insectos acuáticos (principalmente efemerópteros, quironómidos y tricópteros. D. excelsa y G. guidonia (en ambos tipos de malla tuvieron tasas de descomposición más altas que C. scheberiana, M. bidentata y P. acuminata. La descomposición fue más rápida para todas las especies de hoja con la malla gruesa, resaltando el papel de los descomponedores en este hábitat. Tras 42 días de esta incubación, las densidades totales de invertebrados, efemerópteros y tricópteros fueron mayores en las bolsas con hojas de D. excelsa y G. guidonia, e inferiores en P. acuminata, C. scheberiana y M. bidentata. A mayor densidad de insectos hubo mayor tasa de descomposición. Nuestros resultados indican la importancia de la especie de las hojas y de la presencia de macroinvertebrados en el proceso de descomposición de hojarasca en partes altas de arroyos tropicales.

Native trees show conservative water use relative to invasive trees: results from a removal experiment in a Hawaiian wet forest.

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Cavaleri, Molly A; Ostertag, Rebecca; Cordell, Susan; Sack, Lawren

2014-01-01

While the supply of freshwater is expected to decline in many regions in the coming decades, invasive plant species, often 'high water spenders', are greatly expanding their ranges worldwide. In this study, we quantified the ecohydrological differences between native and invasive trees and also the effects of woody invasive removal on plot-level water use in a heavily invaded mono-dominant lowland wet tropical forest on the Island of Hawaii. We measured transpiration rates of co-occurring native and invasive tree species with and without woody invasive removal treatments. Twenty native Metrosideros polymorpha and 10 trees each of three invasive species, Cecropia obtusifolia, Macaranga mappa and Melastoma septemnervium, were instrumented with heat-dissipation sap-flux probes in four 100 m(2) plots (two invaded, two removal) for 10 months. In the invaded plots, where both natives and invasives were present, Metrosideros had the lowest sap-flow rates per unit sapwood, but the highest sap-flow rates per whole tree, owing to its larger mean diameter than the invasive trees. Stand-level water use within the removal plots was half that of the invaded plots, even though the removal of invasives caused a small but significant increase in compensatory water use by the remaining native trees. By investigating the effects of invasive species on ecohydrology and comparing native vs. invasive physiological traits, we not only gain understanding about the functioning of invasive species, but we also highlight potential water-conservation strategies for heavily invaded mono-dominant tropical forests worldwide. Native-dominated forests free of invasive species can be conservative in overall water use, providing a strong rationale for the control of invasive species and preservation of native-dominated stands.

Fatores de virulência de Bacillus thuringiensis: o que existe além das proteínas Cry

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Gislayne Vilas-Bôas

2012-03-01

Virulence Factors of Bacillus thuringiensis Berliner: Something Beyond of Cry Proteins? Abstract. The Cry proteins produced by the entomopathogenic bacterium Bacillus thuringiensis Berliner are widely known due to its high toxicity against a variety of insects. The mode of action of these proteins is specific and becomes B. thuringiensis-based products the most used in biological control programs of insect pests in agriculture and of important human disease vectors. However, while the Cry proteins are the best-known insect-specific virulence factor, strains of B. thuringiensis show also a wide range of other virulence factors, which allow the bacteria to achieve the hemolymph and colonize efficiently the insect host. Among these factors, we highlight the Vip proteins, Cyt, enterotoxins, hemolysins, phospholipases, proteases and enzymes of degradation, in addition to the recently described parasporin. This review explores the action of these virulence factors, as well as, the characterization and control of expression of their genes. Additionally, we discuss aspects related to the ecological niche of the bacteria with emphasis on the characteristics involved in the biosafety of the use of B. thuringiensis-based products for biological control of target insects.

Overexpression of host plant urease in transgenic silkworms.

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Jiang, Liang; Huang, Chunlin; Sun, Qiang; Guo, Huizhen; Peng, Zhengwen; Dang, Yinghui; Liu, Weiqiang; Xing, Dongxu; Xu, Guowen; Zhao, Ping; Xia, Qingyou

2015-06-01

Bombyx mori and mulberry constitute a model of insect-host plant interactions. Urease hydrolyzes urea to ammonia and is important for the nitrogen metabolism of silkworms because ammonia is assimilated into silk protein. Silkworms do not synthesize urease and acquire it from mulberry leaves. We synthesized the artificial DNA sequence ureas using the codon bias of B. mori to encode the signal peptide and mulberry urease protein. A transgenic vector that overexpresses ure-as under control of the silkworm midgut-specific P2 promoter was constructed. Transgenic silkworms were created via embryo microinjection. RT-PCR results showed that urease was expressed during the larval stage and qPCR revealed the expression only in the midgut of transgenic lines. Urea concentration in the midgut and hemolymph of transgenic silkworms was significantly lower than in a nontransgenic line when silkworms were fed an artificial diet. Analysis of the daily body weight and food conversion efficiency of the fourth and fifth instar larvae and economic characteristics indicated no differences between transgenic silkworms and the nontransgenic line. These results suggested that overexpression of host plant urease promoted nitrogen metabolism in silkworms.

Thermolysin damages animal life through degradation of plasma proteins enhanced by rapid cleavage of serpins and activation of proteases.

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Kong, Lulu; Lu, Anrui; Guan, Jingmin; Yang, Bing; Li, Muwang; Hillyer, Julián F; Ramarao, Nalini; Söderhäll, Kenneth; Liu, Chaoliang; Ling, Erjun

2015-01-01

Thermolysin, a metallopeptidase secreted by pathogenic microbes, is concluded as an important virulence factor due to cleaving purified host proteins in vitro. Using the silkworm Bombyx mori as a model system, we found that thermolysin injection into larvae induces the destruction of the coagulation response and the activation of hemolymph melanization, which results in larval death. Thermolysin triggers the rapid degradation of insect and mammalian plasma proteins at a level that is considerably greater than expected in vitro and/or in vivo. To more specifically explore the mechanism, thermolysin-induced changes to key proteins belonging to the insect melanization pathway were assessed as a window for observing plasma protein cleavage. The application of thermolysin induced the rapid cleavage of the melanization negative regulator serpin-3, but did not directly activate the melanization rate-limiting enzyme prophenoloxidase (PPO) or the terminal serine proteases responsible for PPO activation. Terminal serine proteases of melanization are activated indirectly after thermolysin exposure. We hypothesize that thermolysin induces the rapid degradation of serpins and the activation of proteases directly or indirectly, boosting uncontrolled plasma protein degradation in insects and mammalians. © 2014 Wiley Periodicals, Inc.

Biofuel cell backpacked insect and its application to wireless sensing.

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Shoji, Kan; Akiyama, Yoshitake; Suzuki, Masato; Nakamura, Nobuhumi; Ohno, Hiroyuki; Morishima, Keisuke

2016-04-15

This study investigated an enzymatic biofuel cell (BFC) which can be backpacked by cockroaches. The BFC generates electric power from trehalose in insect hemolymph by the trehalase and glucose dehydrogenase (GDH) reaction systems which dehydrogenate β-glucose obtained by hydrolyzing trehalose. First, an insect-mountable BFC (imBFC) was designed and fabricated with a 3D printer. The electrochemical reaction of anode-modified poly-L-lysine, vitamin K3, diaphorase, nicotinamide adenine dinucleotide, GDH and poly(sodium 4-styrenesulfonate) in the imBFC was evaluated and an oxidation current of 1.18 mAcm(-2) (at +0.6 V vs. Ag|AgCl) was observed. Then, the performance of the imBFC was evaluated and a maximum power output of 333 μW (285 μW cm(-)(2)) (at 0.5 V) was obtained. Furthermore, driving of both an LED device and a wireless temperature and humidity sensor device were powered by the imBFC. These results indicate that the imBFC has sufficient potential as a battery for novel ubiquitous robots such as insect cyborgs. Copyright © 2015 Elsevier B.V. All rights reserved.

Genomic organization, sequence characterization and expression analysis of Tenebrio molitor apolipophorin-III in response to an intracellular pathogen, Listeria monocytogenes.

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Noh, Ju Young; Patnaik, Bharat Bhusan; Tindwa, Hamisi; Seo, Gi Won; Kim, Dong Hyun; Patnaik, Hongray Howrelia; Jo, Yong Hun; Lee, Yong Seok; Lee, Bok Luel; Kim, Nam Jung; Han, Yeon Soo

2014-01-25

Apolipophorin III (apoLp-III) is a well-known hemolymph protein having a functional role in lipid transport and immune response of insects. We cloned full-length cDNA encoding putative apoLp-III from larvae of the coleopteran beetle, Tenebrio molitor (TmapoLp-III), by identification of clones corresponding to the partial sequence of TmapoLp-III, subsequently followed with full length sequencing by a clone-by-clone primer walking method. The complete cDNA consists of 890 nucleotides, including an ORF encoding 196 amino acid residues. Excluding a putative signal peptide of the first 20 amino acid residues, the 176-residue mature apoLp-III has a calculated molecular mass of 19,146Da. Genomic sequence analysis with respect to its cDNA showed that TmapoLp-III was organized into four exons interrupted by three introns. Several immune-related transcription factor binding sites were discovered in the putative 5'-flanking region. BLAST and phylogenetic analyses reveal that TmapoLp-III has high sequence identity (88%) with Tribolium castaneum apoLp-III but shares little sequence homologies (molitor. Copyright © 2013 Elsevier B.V. All rights reserved.

A role for the adult fat body in Drosophila male courtship behavior.

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Anna A Lazareva

2007-01-01

Full Text Available Mating behavior in Drosophila depends critically on the sexual identity of specific regions in the brain, but several studies have identified courtship genes that express products only outside the nervous system. Although these genes are each active in a variety of non-neuronal cell types, they are all prominently expressed in the adult fat body, suggesting an important role for this tissue in behavior. To test its role in male courtship, fat body was feminized using the highly specific Larval serum protein promoter. We report here that the specific feminization of this tissue strongly reduces the competence of males to perform courtship. This effect is limited to the fat body of sexually mature adults as the feminization of larval fat body that normally persists in young adults does not affect mating. We propose that feminization of fat body affects the synthesis of male-specific secreted circulating proteins that influence the central nervous system. In support of this idea, we demonstrate that Takeout, a protein known to influence mating, is present in the hemolymph of adult males but not females and acts as a secreted protein.

Common Virulence Factors and Tissue Targets of Entomopathogenic Bacteria for Biological Control of Lepidopteran Pests

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Anaïs Castagnola

2014-01-01

Full Text Available This review focuses on common insecticidal virulence factors from entomopathogenic bacteria with special emphasis on two insect pathogenic bacteria Photorhabdus (Proteobacteria: Enterobacteriaceae and Bacillus (Firmicutes: Bacillaceae. Insect pathogenic bacteria of diverse taxonomic groups and phylogenetic origin have been shown to have striking similarities in the virulence factors they produce. It has been suggested that the detection of phage elements surrounding toxin genes, horizontal and lateral gene transfer events, and plasmid shuffling occurrences may be some of the reasons that virulence factor genes have so many analogs throughout the bacterial kingdom. Comparison of virulence factors of Photorhabdus, and Bacillus, two bacteria with dissimilar life styles opens the possibility of re-examining newly discovered toxins for novel tissue targets. For example, nematodes residing in the hemolymph may release bacteria with virulence factors targeting neurons or neuromuscular junctions. The first section of this review focuses on toxins and their context in agriculture. The second describes the mode of action of toxins from common entomopathogens and the third draws comparisons between Gram positive and Gram negative bacteria. The fourth section reviews the implications of the nervous system in biocontrol.

Induced Hyperproteinemia and Its Effects on the Remodeling of Fat Bodies in Silkworm, Bombyx mori

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Chen, Xue-Dong; Wang, Yong-Feng; Wang, Yu-Long; Li, Qiu-Ying; Ma, Huan-Yu; Wang, Lu; Sima, Yang-Hu; Xu, Shi-Qing

2018-01-01

Hyperproteinemia, which is characterized by an abnormally elevated plasma protein concentration (PPC), is a high-mortality, metabolic complication associated with severe liver and kidney disease. It is difficult to clinically distinguish the difference between the impacts of primary diseases and hyperproteinemia on tissues and organs, and there are no available animal models of hyperproteinemia. Here, we constructed an animal model of hyperproteinemia with a controllable PPC and no primary disease effects in the silkworm Bombyx mori that has attracted interest owing to its potential use in the pathological analysis of model animals. Silkworm have an open circulatory system in which each organ is directly immersed in hemolymph. The fat body (FB) of a silkworm, as a major organ for nutrient storage and energy metabolism, can effectively reflect hyperproteinemia-induced metabolic abnormalities in damaged visceral tissues. A pathogenesis study showed that hyperproteinemia attenuated cell autophagy and apoptosis by attenuating an endocrine hormone, thereby preventing FB remodeling during metamorphosis. Meanwhile, hyperproteinemia increased oxidative stress in the FB and resulted in a dysfunction of amino acid conversion. Supplementation with exogenous 20-hydroxyecdysone effectively mitigated the hyperproteinemia-mediated inhibition of FB remodeling. PMID:29651251

Induced Hyperproteinemia and Its Effects on the Remodeling of Fat Bodies in Silkworm, Bombyx mori

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Xue-Dong Chen

2018-03-01

Full Text Available Hyperproteinemia, which is characterized by an abnormally elevated plasma protein concentration (PPC, is a high-mortality, metabolic complication associated with severe liver and kidney disease. It is difficult to clinically distinguish the difference between the impacts of primary diseases and hyperproteinemia on tissues and organs, and there are no available animal models of hyperproteinemia. Here, we constructed an animal model of hyperproteinemia with a controllable PPC and no primary disease effects in the silkworm Bombyx mori that has attracted interest owing to its potential use in the pathological analysis of model animals. Silkworm have an open circulatory system in which each organ is directly immersed in hemolymph. The fat body (FB of a silkworm, as a major organ for nutrient storage and energy metabolism, can effectively reflect hyperproteinemia-induced metabolic abnormalities in damaged visceral tissues. A pathogenesis study showed that hyperproteinemia attenuated cell autophagy and apoptosis by attenuating an endocrine hormone, thereby preventing FB remodeling during metamorphosis. Meanwhile, hyperproteinemia increased oxidative stress in the FB and resulted in a dysfunction of amino acid conversion. Supplementation with exogenous 20-hydroxyecdysone effectively mitigated the hyperproteinemia-mediated inhibition of FB remodeling.

The neonicotinoids thiacloprid, imidacloprid, and clothianidin affect the immunocompetence of honey bees (Apis mellifera L.).

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Brandt, Annely; Gorenflo, Anna; Siede, Reinhold; Meixner, Marina; Büchler, Ralph

2016-03-01

A strong immune defense is vital for honey bee health and colony survival. This defense can be weakened by environmental factors that may render honey bees more vulnerable to parasites and pathogens. Honey bees are frequently exposed to neonicotinoid pesticides, which are being discussed as one of the stress factors that may lead to colony failure. We investigated the sublethal effects of the neonicotinoids thiacloprid, imidacloprid, and clothianidin on individual immunity, by studying three major aspects of immunocompetence in worker bees: total hemocyte number, encapsulation response, and antimicrobial activity of the hemolymph. In laboratory experiments, we found a strong impact of all three neonicotinoids. Thiacloprid (24h oral exposure, 200 μg/l or 2000 μg/l) and imidacloprid (1 μg/l or 10 μg/l) reduced hemocyte density, encapsulation response, and antimicrobial activity even at field realistic concentrations. Clothianidin had an effect on these immune parameters only at higher than field realistic concentrations (50-200 μg/l). These results suggest that neonicotinoids affect the individual immunocompetence of honey bees, possibly leading to an impaired disease resistance capacity. Copyright © 2016 Elsevier Ltd. All rights reserved.

Hostplant suitability and defensive chemistry of the Catalpa sphinx, Ceratomia catalpae.

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Bowers, M Deane

2003-10-01

The growth and survival of the Catalpa sphinx, Ceratomia catalpae (Sphingidae), were measured on five different species of Catalpa: C. bignonioides, C. bungeii, C. fargeseii, C. ovata, and C. speciosa. Larval growth varied significantly among these host plant species; however, survival did not differ. Quantification of the iridoid glycoside content of larvae, pupae, adults, larval frass, and leaves of the larval host plant, C. bignonioides, by gas chromatography showed that leaves contained both catalpol and catalposide; larvae, pupae, and frass contained only catalpol; and the adults contained no detectable iridoid glycosides. Amounts were highest in the larvae and declined in the pupal stage. Very small amounts of catalpol were detected in adults of the parasitoid, Cotesia congregata, and in the silken cocoons. The hemolymph in which the parasitoid larvae grew contained over 50% dry weight catalpol. Larvae of C. catalpae often regurgitate when disturbed. This may serve as a defense against predators. A comparison of the growth of larvae pinched with forceps to induce regurgitation with those that were not so treated showed that larvae that were pinched, and usually regurgitated, grew significantly more slowly than those that were not.

[Knockdown of InR gene in ventral nephrocytes promotes resistance to toxic stress in Drosophila melanogaster females].

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Andreenkova, O V; Karpova, E K; Menshanov, P N; Rauschenbach, I Yu

2015-02-01

Hemolymph filtration in insects is performed by nephrocytes, additional cells of the circulatory system that are not connected to Malpighian vessels. Drosophila has two types of nephrocytes: the ventral ("garland"), which are situated around the connection site of the esophagus and proventriculus, and the pericardial, which are localized around the heart. In this study, we examined the role of the of insulin-like receptor (InR)gene in regulation of the function of ventral nephrocytes (VNC) in D. melanogaster females. Immunofluorescent analysis of female VNC with anti-InR antibodies revealed for the first time that the InR gene is expressed in VNC cells. To determine whether a change in the level of InR expression has an effect on VNC function in Drosophila females, we implemented an antisense suppressor of the InR gene, together with a driver that is expressed specifically in VNC. VNC function was evaluated by survival of the females exposed to toxic stress (treatment with AgNO3). This study has shown for the first time that suppression of InR expression in VNC leads to a rise in the survival of flies under conditions of toxic stress.

Genotoxic effect of polycyclic aromatic hydrocarbons in the metropolitan area of Porto Alegre, Brazil, evaluated by Helix aspersa (Mueller, 1774)

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Ianistcki, M. [Laboratorio de Genetica Toxicologica, Department of Biology, ULBRA, Av. Farroupilha 8001, Pr. 14/Sala 218, Bairro Sao Jose, CEP 92425-900 Canoas, RS (Brazil); Dallarosa, J. [Laboratorio de Ecologia, UFRGS (Brazil); Sauer, C.; Teixeira, C.E. [Fundacao Estadual de Protecao Ambiental Henrique Luis Roessler, FEPAM, RS (Brazil); Silva, J. da, E-mail: juliana.silva@ulbra.b [Laboratorio de Genetica Toxicologica, Department of Biology, ULBRA, Av. Farroupilha 8001, Pr. 14/Sala 218, Bairro Sao Jose, CEP 92425-900 Canoas, RS (Brazil)

2009-07-15

The purpose of this study was to biomonitor metropolitan areas of Porto Alegre (Brazil) for PAHs associated with atmospheric particles and check their effects on the DNA of the land mollusk Helix aspersa. The sampling sites are located in an urban area with heavy traffic: (i) Canoas, (ii) Sapucaia do Sul, and (iii) FIERGS/Porto Alegre. The samples were collected during a continuous period of 24 hours during 15 days using Stacked Filter Units (SFU) on polycarbonate filters (two separated size fractions: PM{sub 10-2.5} and PM{sub <2.5}). The concentrations of 16 major PAHs were determined according to EPA. Comet assay on H. aspersa hemolymph cells was chosen for genotoxicity evaluation. This evaluation shows that, in general, the smaller PM-size fractions (PM{sub <2.5}) have the highest genotoxicity and contain higher concentrations of extractable organic matter. In addition, associations between chemical characteristics and PM carcinogenicity tend to be stronger for the smaller PM-size fractions. - DNA damage in H. aspersa exposed to atmospheric particulate in Metropolitan Area of Porto Alegre demonstrated association with PAHs in the fine filter (PM{sub <2.5}).

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María Barranco

2015-10-01

The decline in the number of hives of Apis mellifera in many countries of the European Union and the United States has worried beekeepers, farmers, environmentalists, governments and corporations. Among the factors considered are: poor nutrition of bees, artificial supplements and to a lesser extent to a decrease in the abundance and diversity of flowers in the wild plant species all of which makes bees susceptible to diseases caused by viruses, bacteria and microsporidia which, in turn, are considered cofactors for the Colony Collapse Disorder (CCD. It also noted that the mites as Acarapis woodi and Varroa destructor may be causing the CCD by feeding on the hemolymph of larvae and adults thus lowering their defenses and favoring the spread of infectious diseases. Another hypothesis points to a general weakening in bee populations due to low genetic diversity by producing large amounts of colonies from a restricted set of queen bees and artificial insemination of the same. But of all the above factors, pesticides, primarily the so-called neonicotinoids, are considered as one of the main causes of the disappearance of bees worldwide in recent years. In this article we examine the evidence about

Drosophila comet assay: insights, uses, and future perspectives

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Gaivão, Isabel; Sierra, L. María

2014-01-01

The comet assay, a very useful tool in genotoxicity and DNA repair testing, is being applied to Drosophila melanogaster since around 15 years ago, by several research groups. This organism is a valuable model for all kind of processes related to human health, including DNA damage response. The assay has been performed mainly in vivo using different larvae cell types (from brain, midgut, hemolymph, and imaginal disk), but also in vitro with the S2 cell line. Since its first application, it has been used to analyze the genotoxicity and action mechanisms of different chemicals, demonstrating good sensitivity and proving its usefulness. Moreover, it is the only assay that can be used to analyze DNA repair in somatic cells in vivo, comparing the effects of chemicals in different repair strains, and to quantitate repair activities in vitro. Additionally, the comet assay in Drosophila, in vivo and in vitro, has been applied to study the influence of protein overexpression on genome integrity and degradation. Although the assay is well established, it could benefit from some research to determine optimal experimental design to standardize it, and then to allow comparisons among laboratories independently of the chosen cell type. PMID:25221574

[Identification of novel therapeutically effective antibiotics using silkworm infection model].

Science.gov (United States)

Hamamoto, Hiroshi; Urai, Makoto; Paudel, Atmika; Horie, Ryo; Murakami, Kazuhisa; Sekimizu, Kazuhisa

2012-01-01

Most antibiotics obtained by in vitro screening with antibacterial activity have inappropriate properties as medicines due to their toxicity and pharmacodynamics in animal bodies. Thus, evaluation of the therapeutic effects of these samples using animal models is essential in the crude stage. Mammals are not suitable for therapeutic evaluation of a large number of samples due to high costs and ethical issues. We propose the use of silkworms (Bombyx mori) as model animals for screening therapeutically effective antibiotics. Silkworms are infected by various pathogenic bacteria and are effectively treated with similar ED(50) values of clinically used antibiotics. Furthermore, the drug metabolism pathways, such as cytochrome P450 and conjugation systems, are similar between silkworms and mammals. Silkworms have many advantages compared with other infection models, such as their 1) low cost, 2) few associated ethical problems, 3) adequate body size for easily handling, and 4) easier separation of organs and hemolymph. These features of the silkworm allow for efficient screening of therapeutically effective antibiotics. In this review, we discuss the advantages of the silkworm model in the early stages of drug development and the screening results of some antibiotics using the silkworm infection model.

Extracellular digestion during hyposaline exposure in the Dungeness crab, Cancer magister, and the blue crab, Callinectes sapidus.

Science.gov (United States)

Curtis, Daniel L; van Breukelen, Frank; McGaw, Iain J

2013-12-01

Extracellular digestive processes were examined in the Dungeness crab, Cancer magister and the blue crab, Callinectes sapidus, during hyposaline exposure. Both species are found in estuaries as adults, but vary in their ability to balance the cardiovascular and respiratory demands of concurrent osmoregulation and digestion. The weak osmoregulator, C. magister, is unable to balance the demands of osmoregulation and digestion. Concordant with observed decreases in oxygen consumption and mechanical digestion, proteolytic digestion within the foregut and hepatopancreas was delayed, resulting in a relative reduction of circulating amino acids post-feeding in low salinity. In contrast, the efficient osmoregulator, C. sapidus, balances the demands of osmoregulation and digestion, and mechanical digestion continues unabated in low salinity. Protease activity in the gut fluid and hepatopancreas showed either no change or a reduction over time. The transport of amino acids into the cells post-feeding is opposed by an efflux of amino acids at the cellular level, and resulted in a build up of amino acids in the hemolymph. Despite differences in the extracellular responses to low salinity exposure following feeding, both species were able to maintain high digestive efficiencies. © 2013.

Insecticidal activity of two proteases against Spodoptera frugiperda larvae infected with recombinant baculoviruses

Science.gov (United States)

2010-01-01

Background Baculovirus comprise the largest group of insect viruses most studied worldwide, mainly because they efficiently kill agricutural insect pests. In this study, two recombinant baculoviruses containing the ScathL gene from Sarcophaga peregrina (vSynScathL), and the Keratinase gene from the fungus Aspergillus fumigatus (vSynKerat), were constructed. and their insecticidal properties analysed against Spodoptera frugiperda larvae. Results Bioassays of third-instar and neonate S. frugiperda larvae with vSynScathL and vSynKerat showed a decrease in the time needed to kill the infected insects when compared to the wild type virus. We have also shown that both recombinants were able to increase phenoloxidase activity in the hemolymph of S. frugiperda larvae. The expression of proteases in infected larvae resulted in destruction of internal tissues late in infection, which could be the reason for the increased viral speed of kill. Conclusions Baculoviruses and their recombinant forms constitute viable alternatives to chemical insecticides. Recombinant baculoviruses containing protease genes can be added to the list of engineered baculoviruses with great potential to be used in integrated pest management programs. PMID:20587066

Genotoxicity of Nicotiana tabacum leaves on Helix aspersa.

Science.gov (United States)

da Silva, Fernanda R; Erdtmann, Bernardo; Dalpiaz, Tiago; Nunes, Emilene; Ferraz, Alexandre; Martins, Tales L C; Dias, Johny F; da Rosa, Darlan P; Porawskie, Marilene; Bona, Silvia; da Silva, Juliana

2013-07-01

Tobacco farmers are routinely exposed to complex mixtures of inorganic and organic chemicals present in tobacco leaves. In this study, we examined the genotoxicity of tobacco leaves in the snail Helix aspersa as a measure of the risk to human health. DNA damage was evaluated using the micronucleus test and the Comet assay and the concentration of cytochrome P450 enzymes was estimated. Two groups of snails were studied: one fed on tobacco leaves and one fed on lettuce (Lactuca sativa L) leaves (control group). All of the snails received leaves (tobacco and lettuce leaves were the only food provided) and water ad libitum. Hemolymph cells were collected after 0, 24, 48 and 72 h. The Comet assay and micronucleus test showed that exposure to tobacco leaves for different periods of time caused significant DNA damage. Inhibition of cytochrome P450 enzymes occurred only in the tobacco group. Chemical analysis indicated the presence of the alkaloid nicotine, coumarins, saponins, flavonoids and various metals. These results show that tobacco leaves are genotoxic in H. aspersa and inhibit cytochrome P450 activity, probably through the action of the complex chemical mixture present in the plant.

Genotoxicity of Nicotiana tabacum leaves on Helix aspersa

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Fernanda R. da Silva

2013-01-01

Full Text Available Tobacco farmers are routinely exposed to complex mixtures of inorganic and organic chemicals present in tobacco leaves. In this study, we examined the genotoxicity of tobacco leaves in the snail Helix aspersa as a measure of the risk to human health. DNA damage was evaluated using the micronucleus test and the Comet assay and the concentration of cytochrome P450 enzymes was estimated. Two groups of snails were studied: one fed on tobacco leaves and one fed on lettuce (Lactuca sativa L leaves (control group. All of the snails received leaves (tobacco and lettuce leaves were the only food provided and water ad libitum. Hemolymph cells were collected after 0, 24, 48 and 72 h. The Comet assay and micronucleus test showed that exposure to tobacco leaves for different periods of time caused significant DNA damage. Inhibition of cytochrome P450 enzymes occurred only in the tobacco group. Chemical analysis indicated the presence of the alkaloid nicotine, coumarins, saponins, flavonoids and various metals. These results show that tobacco leaves are genotoxic in H. aspersa and inhibit cytochrome P450 activity, probably through the action of the complex chemical mixture present in the plant.

Hypoxia increases susceptibility of Pacific white shrimp to whitespot syndrome virus (WSSV

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M. Lehmann

2016-04-01

Full Text Available The present study aimed to evaluate the mortality, reactive oxygen species production (ROS and total hemocyte counts (THC of the marine shrimp Litopenaeus vannamei infected with the white spot syndrome virus (WSSV at three levels of oxygen saturation. For this, 360 shrimp (20±2g were distributed in 24 tanks (60L, divided in two groups (infected and non-infected, which were subjected to 30, 60 and 100% of dissolved oxygen saturation (in quadruplicate. During 96 hours after infection, daily hemolymph samples were collected for hemato-immunological parameter evaluation (THC and ROS and dead animals were removed and computed to assess cumulative mortality rates. In the infected group, animals subjected to 100% saturation showed higher ROS production (P<0.05 after 48 hours, while THC was significantly reduced (P<0.05, regardless of oxygen saturation. The hypoxia resulted in high mortality when compared to 100% saturation condition. In the uninfected group, no significant differences were observed in all evaluated parameters. Thus, the hypoxia condition increased the susceptibility of shrimp to the infection of WSSV, which may be partly related to the low ROS production showed by the animals subjected to 30% oxygen saturation.

The spread of Vibrio parahaemolyticus in tissues of the Pacific white shrimp Litopenaeus vannamei analyzed by PCR and histopathology.

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Khimmakthong, Umaporn; Sukkarun, Pimwarang

2017-12-01

V. parahaemolyticus are bacteria that cause the Acute Hepatopancreatic Necrosis Disease (AHPND), or Early Mortality Syndrome (EMS), in shrimp. To further understand the pathogenesis mechanisms of V. parahaemolyticus infection in shrimp, the spreading of this bacterium in various tissues was investigated. The spread of infection in shrimp that were exposed to seawater bacteria was studied by PCR and histopathology at 1 min, 1, 6, 12, 24, 48 and 72 h after exposure. The PCR results showed that V. parahaemolyticus was at its most widespread at 6 h after exposure, at which point V. parahaemolyticus was found in the gills, hepatopancreas, intestine, muscles, and hemolymph. However, examinations after 6 h of infection found only small amounts of V. parahaemolyticus in hepatopancreas and intestines. Histopathology of the hepatopancreas showed abnormalities on gross examination at 1 min-72 h after exposure. This study indicates that V. parahaemolyticus can spread quickly by using the hepatopancreas as the target tissue. After 6 h of infection, V. parahaemolyticus was eliminated by immune system while their toxins still caused damage to shrimp tissues. Copyright © 2017 Elsevier Ltd. All rights reserved.

The characterization of the fat bodies and oenocytes in the adult females of the sand fly vectors Lutzomyia longipalpis and Phlebotomus papatasi.

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de Assis, Wiviane Alves; Malta, Juliana; Pimenta, Paulo Filemon P; Ramalho-Ortigão, José Marcelo; Martins, Gustavo Ferreira

2014-09-01

The fat body (FB) is responsible for the storage and synthesis of the majority of proteins and metabolites secreted into the hemolymph. Oenocytes are responsible for lipid processing and detoxification. The FB is distributed throughout the insect body cavity and organized as peripheral and perivisceral portions in the abdomen, with trophocytes and oenocytes attached to the peripheral portion. Here, we investigated the morphology and the subcellular changes in the peripheral and perivisceral FBs and in oenocytes of the sand flies Lutzomyia longipalpis and Phlebotomus papatasi after blood feeding. In L. longipalpis two-sized oenocytes (small and large) were identified, with both cell types displaying well-developed reticular system and smooth endoplasmic reticulum, whereas in P. papatasi, only small cells were observed. Detailed features of FBs of L. longipalpis and P. papatasi are shared either prior to or after blood feeding. The peripheral and perivisceral FBs responded to blood feeding with the development of glycogen zones and rough endoplasmic reticulum. This study provides the first detailed description of the FBs and oenocytes in sand flies, contributing significantly towards are better understanding of the biology of such important disease vectors. Copyright © 2014 Elsevier Ltd. All rights reserved.

Rove beetles of medical importance in Brazil (Coleoptera, Staphylinidae, Paederinae

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Juliana S. Vieira

2014-09-01

Full Text Available Rove beetles of medical importance in Brazil (Coleoptera, Staphylinidae, Paederinae. The rove beetles of the genus Paederus Fabricius, 1775 are the most important group within Coleoptera causing dermatitis around the world. The medical importance of Paederus depends on its toxic hemolymph released when these beetles are crushed on human skin. The effects are mainly dermatitis linearis and some sporadic cases of conjunctivitis. In Brazil seven species of Paederus are known to cause dermatitis: P. amazonicus Sharp, 1876, P. brasiliensis Erichson, 1840, P. columbinus Laporte, 1835, P. ferus Erichson, 1840, P. mutans Sharp, 1876, P. protensus Sharp, 1876 stat. rev., and Paederus rutilicornis Erichson, 1840. Paederus mutans and P. protensus are for the first time recorded as of medical importance, whereas the record of P. rutilicornis in Brazil is doubtful. All seven species are redescribed and a dichotomous key is provided. The geographic distributions of all species are documented. The results provided here include the most recent and relevant taxonomic revision of Paederus of the Neotropical region, the first identification key for Brazilian species and the increase of recorded species of medical importance in the world.

Effects of waterborne nickel on the physiological and immunological parameters of the Pacific abalone Haliotis discus hannai during thermal stress.

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Min, Eun Young; Cha, Yong-Joo; Kang, Ju-Chan

2015-09-01

In this study, the 96-h LC50 at 22 and 26 °C values was 28.591 and 11.761 mg/L, respectively, for NiCl2 exposure in the abalone. The alteration of physiological and immune-toxicological parameters such as the total hemocyte count (THC), lysozyme, phenoloxidase (PO), and phagocytosis activity was measured in the abalone exposed to nickel (200 and 400 μg/L) under thermal stress for 96 h. In this study, Mg and THC decreased, while Ca, lysozyme, PO, and phagocytosis activity increased in the hemolymph of Pacific abalone exposed to NiCl2 when compared to a control at both 22 and 26 °C. However, these parameters were not affected by a rise in temperature from 22 to 26 °C in non-exposed groups. Our results showed that NiCl2 below 400 μg/L was able to stimulate immune responses in abalone. However, complex stressors, thermal changes, or NiCl2 can modify the immunological response and lead to changes in the physiology of host-pollutant interactions in the abalone.

Effects of Titanium Dioxide Nanoparticles on the Synthesis of Fibroin in Silkworm (Bombyx mori).

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Ni, Min; Li, FanChi; Tian, JiangHai; Hu, JingSheng; Zhang, Hua; Xu, KaiZun; Wang, BinBin; Li, YangYang; Shen, WeiDe; Li, Bing

2015-08-01

Silkworm (Bombyx mori) is an economically important insect, and its silk production capacity largely depends on its ability to synthesize fibroin. While breeding of B. mori varieties has been a key strategy to improve silk production, little improvement of B. mori silk production has been achieved to date. As a result, the development of sericulture economy has not progressed well, pointing to the need of new ways for improvement of B. mori silk production. Titanium dioxide nanoparticles (TiO2 NPs), a food additive widely used for livestock, have been shown to promote animal growth and increase the protein synthesis in animals. However, no studies on effect of TiO2 NPs on fibroin synthesis in B. mori have been available. In this study, the differential expression profiles of genes and proteins in the silk gland of B. mori fed without or with TiO2 NPs (5 μg ml(-1)) were analyzed and compared using digital gene expression (DGE), reverse transcription quantitative polymerase chain reaction (RT-qPCR), semi-qPCR, and Western blot analysis. The effects of TiO2 NPs feeding on the activity of proteases in the midgut and the synthesis and transportation of amino acids in hemolymph were also investigated. DGE analyses showed that among a total of 4,741 genes detected, 306 genes were differentially expressed after the TiO2 NPs feeding, of which 137 genes were upregulated whereas 169 genes were downregulated. 106 genes were shown to be involved in fibroin synthesis, of which 97 genes, including those encoding cuticular protein glycine-rich 10, serine protease inhibitor 28, aspartate aminotransferase, lysyl-tRNA synthetase, and splicing factor arginine/serine-rich 6, and silk gland factor-1 (SGF-1), were upregulated with the maximum induction of 8.52-folds, whereas nine genes, including those encoding aspartylglucosaminidase, the cathepsin L in Tribolium castaneum, and similar to SPRY domain-containing SOCS box protein 3, were downregulated with the maximum reduction of 8

Developmental changes in concentrations of vitellin, vitellogenin, and lipids in hemolymph, hepatopancreas, and ovaries from different ovarian stages of Indian white prawn Fenneropenaeus indicus

DEFF Research Database (Denmark)

Boucard, C.G.V.; Levy, P.; Ceccaldi, H.J.

2002-01-01

The objective of the present study was to characterize the relationship between vitellogenin (Vtg) and vitellin (Vt) concentration profiles during the reproductive cycle of the penaeid prawn Fenneropenaeus indicus. Vt was purified from ovaries of vitellogenic females by gradient ultracentrifugati...

Thermal acclimation mitigates cold-induced paracellular leak from the Drosophila gut.

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MacMillan, Heath A; Yerushalmi, Gil Y; Jonusaite, Sima; Kelly, Scott P; Donini, Andrew

2017-08-18

Chill susceptible insects suffer tissue damage and die at low temperatures. The mechanisms that cause chilling injury are not well understood but a growing body of evidence suggests that a cold-induced loss of ion and water homeostasis leads to hemolymph hyperkalemia that depolarizes cells, leading to cell death. The apparent root of this cascade is the net leak of osmolytes down their concentration gradients in the cold. Many insects, however, are capable of adjusting their thermal physiology, and cold-acclimated Drosophila can maintain homeostasis and avoid injury better than warm-acclimated flies. Here, we test whether chilling causes a loss of epithelial barrier function in female adult Drosophila, and provide the first evidence of cold-induced epithelial barrier failure in an invertebrate. Flies had increased rates of paracellular leak through the gut epithelia at 0 °C, but cold acclimation reduced paracellular permeability and improved cold tolerance. Improved barrier function was associated with changes in the abundance of select septate junction proteins and the appearance of a tortuous ultrastructure in subapical intercellular regions of contact between adjacent midgut epithelial cells. Thus, cold causes paracellular leak in a chill susceptible insect and cold acclimation can mitigate this effect through changes in the composition and structure of transepithelial barriers.

White Shrimp Litopenaeus vannamei That Have Received Gracilaria tenuistipitata Extract Show Early Recovery of Immune Parameters after Ammonia Stressing

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Yu-Yuan Chen

2015-06-01

Full Text Available White shrimp Litopenaeus vannamei immersed in seawater (35‰ containing Gracilaria tenuistipitata extract (GTE at 0 (control, 400, and 600 mg/L for 3 h were exposed to 5 mg/L ammonia-N (ammonia as nitrogen, and immune parameters including hyaline cells (HCs, granular cells (GCs, including semi-granular cells, total hemocyte count (THC, phenoloxidase (PO activity, respiratory bursts (RBs, superoxide dismutase (SOD activity, lysozyme activity, and hemolymph protein level were examined 24~120 h post-stress. The immune parameters of shrimp immersed in 600 mg/L GTE returned to original values earlier, at 96~120 h post-stress, whereas in control shrimp they did not. In another experiment, shrimp were immersed in seawater containing GTE at 0 and 600 mg/L for 3 h and examined for transcript levels of immune-related genes at 24 h post-stress. Transcript levels of lipopolysaccharide and β-1,3-glucan binding protein (LGBP, peroxinectin (PX, cytMnSOD, mtMnSOD, and HSP70 were up-regulated at 24 h post-stress in GTE receiving shrimp. We concluded that white shrimp immersed in seawater containing GTE exhibited a capability for maintaining homeostasis by regulating cellular and humoral immunity against ammonia stress as evidenced by up-regulated gene expression and earlier recovery of immune parameters.

Toxic Effects of Couroupita guianensis Against Spodoptera litura (Fabricius) (Lepidoptera: Noctuidae).

Science.gov (United States)

Baskar, K; Ignacimuthu, S; Jayakumar, M

2015-02-01

Laboratory experiments were conducted to find out the efficacy of different crude extracts and fractions of Couroupita guianensis (Lecythidaceae) against Spodoptera litura (Fabricius). Results revealed that hexane, chloroform and ethyl acetate extracts of C. guianensis showed larvicidal and pupicidal activities against S. litura. Maximum larvicidal activity (68.66%) was observed in hexane extract at 5.0% concentration followed by chloroform and ethyl acetate extracts, and least LC50 value of 2.64% was observed in hexane extract. A 100% pupicidal activity was observed in hexane extract. Based on the efficacy of crude extracts, the effective crude extract (hexane extract) was further fractionated and subjected to screening for biological activities against S. litura. Among the eight fractions isolated from the hexane extract, fraction 8 showed maximum antifeedant activity (81. 8%) and larvicidal activity (76.9%) at 1000-ppm concentration; this fraction showed least LC50 value of 375.92 ppm for larvicidal activity. Cent per cent pupicidal activity was recorded. Reduced midgut and hemolymph protein contents were observed at 1000 ppm of fraction 8. Histopathological studies revealed that fraction 8 severely damaged the midgut cells of S. litura. This fraction could be used to develop botanical formulation to control agricultural pests.

Altered dopamine levels induced by the parasite Profilicollis antarcticus on its intermediate host, the crab Hemigrapsus crenulatus

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JOSÉ MIGUEL ROJAS

2005-01-01

Full Text Available A serotonergic pathway is apparently involved in parasite-host interactions. Previous studies conducted in our laboratory showed increased rates in oxygen consumption and alterations in body posture in the crab Hemigrapsus crenulatus parasitized by the acanthocephalan, Profilicollis antarcticus. Such changes may be related to the functions described for biogenic amines in crustaceans. During the infective stage the acanthocephalans live freely in the hemocelomic cavity, suggesting that the possible alteration induced by biogenic amines may be related to their neurohormonal function in crustaceans. To test whether the presence of P. antarcticus produced neurohormonal changes in its intermediate host, H. crenulatus, we analyzed serotonin and dopamine levels in the host using HPLC with electrochemical detection. Two groups of 11 female crabs were studied; one group was artificially inoculated with two cystacanths while the other was used as the control. Our results show a dramatic increase in hemolymph dopamine, but not serotonin in H. crenulatus parasitized by the acanthocephalan P. antarcticus. Our results, along with those reported by Maynard (1996, suggest a parasite-specific strategy involved in the behavior alteration caused by the acanthocephalans on their intermediate host. The use of a biogenic amine as a mechanism of interaction by the parasites gives them an endless number of alternative potential actions on their intermediate hosts

Altered dopamine levels induced by the parasite Profilicollis antarcticus on its intermediate host, the crab Hemigrapsus crenulatus.

Science.gov (United States)

Rojas, José Miguel; Ojeda, F Patricio

2005-01-01

A serotonergic pathway is apparently involved in parasite-host interactions. Previous studies conducted in our laboratory showed increased rates in oxygen consumption and alterations in body posture in the crab Hemigrapsus crenulatus parasitized by the acanthocephalan, Profilicollis antarcticus. Such changes may be related to the functions described for biogenic amines in crustaceans. During the infective stage the acanthocephalans live freely in the hemocelomic cavity, suggesting that the possible alteration induced by biogenic amines may be related to their neurohormonal function in crustaceans. To test whether the presence of P. antarcticus produced neurohormonal changes in its intermediate host, H. crenulatus, we analyzed serotonin and dopamine levels in the host using HPLC with electrochemical detection. Two groups of 11 female crabs were studied; one group was artificially inoculated with two cystacanths while the other was used as the control. Our results show a dramatic increase in hemolymph dopamine, but not serotonin in H. crenulatus parasitized by the acanthocephalan P. antarcticus. Our results, along with those reported by Maynard (1996), suggest a parasite-specific strategy involved in the behavior alteration caused by the acanthocephalans on their intermediate host. The use of a biogenic amine as a mechanism of interaction by the parasites gives them an endless number of alternative potential actions on their intermediate hosts.

Gonadotropic and physiological functions of juvenile hormone in Bumblebee (Bombus terrestris workers.

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Hagai Shpigler

Full Text Available The evolution of advanced sociality in bees is associated with apparent modifications in juvenile hormone (JH signaling. By contrast to most insects in which JH is a gonadotropin regulating female fertility, in the highly eusocial honey bee (Apis mellifera JH has lost its gonadotrophic function in adult females, and instead regulates age-related division of labor among worker bees. In order to shed light on the evolution of JH signaling in bees we performed allatectomy and replacement therapies to manipulate JH levels in workers of the "primitively eusocial" bumblebee Bombus terrestris. Allatectomized worker bees showed remarkable reduction in ovarian development, egg laying, Vitellogenin and Krüppel homolog 1 fat body transcript levels, hemolymph Vitellogenin protein abundance, wax secretion, and egg-cell construction. These effects were reverted, at least partially, by treating allatectomized bees with JH-III, the natural JH of bees. Allatectomy also affected the amount of ester component in Dufour's gland secretion, which is thought to convey a social signal relating to worker fertility. These findings provide a strong support for the hypothesis that in contrast to honey bees, JH is a gonadotropin in bumblebees and lend credence to the hypothesis that the evolution of advanced eusociality in honey bees was associated with major modifications in JH signaling.

Facultative parasitism by the bivalve Kurtiella pedroana in the sand crab Emerita analoga

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Bhaduri, Ritin; Valentich-Scott, Paul; Hilgers, Mark; Singh, Rajvir; Hickman, Mikaila; Lafferty, Kevin D.

2017-01-01

It is rare that an organism capable of independent or commensalistic existence can also become endoparasitic on a host. In this study, we documented a potential step toward parasitism in the commensal clam Kurtiella pedroana (Bivalvia: Galeommatoidea). Galeommatoideans are known commensals of various invertebrates, including crustaceans. Emerita analoga (Decapoda: Hippidae) is an abundant intertidal decapod inhabiting sandy beaches of the Pacific coast of North and South America. Crabs collected from Monterey Bay, California, were measured and examined externally and internally for associated molluscs. Out of the 520 crabs, 37 large female individuals harbored 49 bivalves (prevalence of 7.11% and mean intensity of 1.3). Forty-one ectocommensal clams were either inside the crab's branchial chambers or on their lateroventral surfaces, and were attached by byssal threads. Our key finding was eight clams that lacked byssal threads and were living in the hemocoel. These internal clams were significantly smaller than the ectocommensals. Because these internal clams lacked access to their normal food, we hypothesize they might have fed on their host's hemolymph as would a parasite. This clam species likely can't reproduce inside its host, implying that endoparasitism is a dead-end state for K. pedroana. Facultative parasitism in a free-living or an ectocommensal is uncommon and suggests a pathway to parasitism.

Crimean-Congo Hemorrhagic Fever: Tick-Host-Virus Interactions

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Anna Papa

2017-05-01

Full Text Available Crimean-Congo hemorrhagic fever virus (CCHFV is transmitted to humans by bite of infected ticks or by direct contact with blood or tissues of viremic patients or animals. It causes to humans a severe disease with fatality up to 30%. The current knowledge about the vector-host-CCHFV interactions is very limited due to the high-level containment required for CCHFV studies. Among ticks, Hyalomma spp. are considered the most competent virus vectors. CCHFV evades the tick immune response, and following its replication in the lining of the tick's midgut, it is disseminated by the hemolymph in the salivary glands and reproductive organs. The introduction of salivary gland secretions into the host cells is the major route via which CCHFV enters the host. Following an initial amplification at the site of inoculation, the virus is spread to the target organs. Apoptosis is induced via both intrinsic and extrinsic pathways. Genetic factors and immune status of the host may affect the release of cytokines which play a major role in disease progression and outcome. It is expected that the use of new technology of metabolomics, transcriptomics and proteomics will lead to improved understanding of CCHFV-host interactions and identify potential targets for blocking the CCHFV transmission.

Spiny lobsters detect conspecific blood-borne alarm cues exclusively through olfactory sensilla.

Science.gov (United States)

Shabani, Shkelzen; Kamio, Michiya; Derby, Charles D

2008-08-01

When attacked by predators, diverse animals actively or passively release molecules that evoke alarm and related anti-predatory behavior by nearby conspecifics. The actively released molecules are alarm pheromones, whereas the passively released molecules are alarm cues. For example, many insects have alarm-signaling systems that involve active release of alarm pheromones from specialized glands and detection of these signals using specific sensors. Many crustaceans passively release alarm cues, but the nature of the cues, sensors and responses is poorly characterized. Here we show in laboratory and field experiments that injured Caribbean spiny lobsters, Panulirus argus, passively release alarm cues via blood (hemolymph) that induce alarm responses in the form of avoidance and suppression of feeding. These cues are detected exclusively through specific olfactory chemosensors, the aesthetasc sensilla. The alarm cues for Caribbean spiny lobsters are not unique to the species but do show some phylogenetic specificity: P. argus responds primarily with alarm behavior to conspecific blood, but with mixed alarm and appetitive behaviors to blood from the congener Panulirus interruptus, or with appetitive behaviors to blood from the blue crab Callinectes sapidus. This study lays the foundation for future neuroethological studies of alarm cue systems in this and other decapod crustaceans.

Enhanced trace element concentrations in tissues of the clam Ruditapes decussatus transplanted to areas influenced by human activities (Ria Formosa, Portugal

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Maria João Botelho

2017-06-01

Full Text Available To examine the extent to which human activities near the Ria Formosa coastal lagoon influence the accumulation of trace elements (TE in Ruditapes decussatus, individuals were transplanted from a natural bank located in the lower lagoon to three sites located in clam growth grounds under the influence of a small city (Faro, a fish farming centre (Olhão and a site near the lagoon inlet (Lavajo. Concentrations were determined in substrate of the clam grounds and in the digestive gland, gills, mantle plus siphons, and remaining tissues of clams in four periods of the year. These measurements were accomplished with the monthly survey of the gametogenic stages, condition index, proteins, glycogen, total lipids, pH and osmolarity of hemolymph. Arsenic, Cu, Mn, V, Cr and Pb were preferentially linked to the digestive gland, while Cd was linked to the gills. TE concentrations in the digestive gland and remaining tissues were higher in winter, most likely reflecting additional inputs associated with rain. The lack of disruptions in biological parameters and the prolonged period of spawning and gonad recovery in clams suggest that the current TE availability in the lagoon has a minor influence on the reproductive cycle and hence on clam production.

The plasticity of extracellular fluid homeostasis in insects.

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Beyenbach, Klaus W

2016-09-01

In chemistry, the ratio of all dissolved solutes to the solution's volume yields the osmotic concentration. The present Review uses this chemical perspective to examine how insects deal with challenges to extracellular fluid (ECF) volume, solute content and osmotic concentration (pressure). Solute/volume plots of the ECF (hemolymph) reveal that insects tolerate large changes in all three of these ECF variables. Challenges beyond those tolerances may be 'corrected' or 'compensated'. While a correction simply reverses the challenge, compensation accommodates the challenge with changes in the other two variables. Most insects osmoregulate by keeping ECF volume and osmotic concentration within a wide range of tolerance. Other insects osmoconform, allowing the ECF osmotic concentration to match the ambient osmotic concentration. Aphids are unique in handling solute and volume loads largely outside the ECF, in the lumen of the gut. This strategy may be related to the apparent absence of Malpighian tubules in aphids. Other insects can suspend ECF homeostasis altogether in order to survive extreme temperatures. Thus, ECF homeostasis in insects is highly dynamic and plastic, which may partly explain why insects remain the most successful class of animals in terms of both species number and biomass. © 2016. Published by The Company of Biologists Ltd.

Biochemical characterisation of the tissue degrading enzyme, collagenase, in the spined soldier bug, Podisus maculiventris (Hemiptera: Pentatomidae

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Ghamari Mahboob

2014-07-01

Full Text Available Podisus maculiventris (Say is a generalist predator attacking many insect species from different orders. The bug injects saliva into its prey's body. The ingested hemolymph and liquefied internal tissues pass through the bug's alimentary tract. Collagenase working on peptide bonds of collagen and basement membrane proteins, leads to the disintegration of the prey's internal organs. As yet, there is an almost complete lack of knowledge on the collagenase activity in P. maculiventris. The collagenase activity of the salivary glands and midgut was optimum at pH 8.0 which was congruent with the optimal pH of the total proteolytic activity of the salivary glands. More collagenolytic activity was determined in the posterior lobe of the salivary glands and anterior midgut. Significant inhibition of collagenolytic activity by ethylenediaminetetraacetic acid (EDTA revealed the enzyme is a metalloproteinase. The collagenase activity notably decreased when the bug went hungry. The salivary gland collagenase is a vital enzyme in extra-oral digestion and facilitates the action of other digestive enzymes. The midgut collagenase may be involved in the digestion of the ingested muscle fibers. The collagenase probably acts as an intoxicating agent in the saliva (venom of P. maculiventris. Paralysing toxins are present in the salivary gland secretion.

Silencing a sugar transporter gene reduces growth and fecundity in the brown planthopper, Nilaparvata lugens (Stål) (Hemiptera: Delphacidae).

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Ge, Lin-Quan; Jiang, Yi-Ping; Xia, Ting; Song, Qi-Sheng; Stanley, David; Kuai, Peng; Lu, Xiu-Li; Yang, Guo-Qing; Wu, Jin-Cai

2015-07-17

The brown planthopper (BPH), Nilaparvata lugens, sugar transporter gene 6 (Nlst6) is a facilitative glucose/fructose transporter (often called a passive carrier) expressed in midgut that mediates sugar transport from the midgut lumen to hemolymph. The influence of down regulating expression of sugar transporter genes on insect growth, development, and fecundity is unknown. Nonetheless, it is reasonable to suspect that transporter-mediated uptake of dietary sugar is essential to the biology of phloem-feeding insects. Based on this reasoning, we posed the hypothesis that silencing, or reducing expression, of a BPH sugar transporter gene would be deleterious to the insects. To test our hypothesis, we examined the effects of Nlst6 knockdown on BPH biology. Reducing expression of Nlst6 led to profound effects on BPHs. It significantly prolonged the pre-oviposition period, shortened the oviposition period, decreased the number of eggs deposited and reduced body weight, compared to controls. Nlst6 knockdown also significantly decreased fat body and ovarian (particularly vitellogenin) protein content as well as vitellogenin gene expression. Experimental BPHs accumulated less fat body glucose compared to controls. We infer that Nlst6 acts in BPH growth and fecundity, and has potential as a novel target gene for control of phloem-feeding pest insects.

Stress biomarkers and alkali-labile phosphate level in mussels (Mytilus galloprovincialis) collected in the urban area of Venice (Venice Lagoon, Italy)

International Nuclear Information System (INIS)

Pampanin, Daniela M.; Marangon, Ilenia; Volpato, Elisa; Campesan, Giancarlo; Nasci, Cristina

2005-01-01

In this study, a spatial and temporal survey at three sites located in the 'canals' of the Venice historic centre (Italy) and at a reference site was undertaken to evaluate stress effects on mussels sampled in the Venice urban area, where raw sewage is discharged without treatment directly into the water. A battery of biomarkers (metallothionein, micronuclei, condition index and survival in air) was used to evaluate the stress condition of the animals. At the same time the alkali-labile phosphate assay (ALP) was performed in mussel' hemolymph with the aim to find an estrogenic effect biomarker in this mussel species. Biomarker results showed an impairment of the general health condition in the mussels coming from the urban area, in agreement with the chemical analysis. Significantly higher level of the ALP was found in male mussels sampled in April in the urban area, in comparison with the ones from the reference site (P0.001). Finally, the PCA proved an easy and useful tool to summarize the obtained results, also able to classify the data to indicate a pollution gradient in the Venice urban area. - The overall biological and chemical data show a higher stress condition in the mussels (Mytilus galloprovincialis) from the urban area of Venice

Identification and characterization of a pyridoxal 5'-phosphate phosphatase in the silkworm (Bombyx mori).

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Huang, ShuoHao; Han, CaiYun; Ma, ZhenQiao; Zhou, Jie; Zhang, JianYun; Huang, LongQuan

2017-03-01

Vitamin B 6 comprises six interconvertible pyridine compounds, among which pyridoxal 5'-phosphate (PLP) is a coenzyme for over 140 enzymes. PLP is also a very reactive aldehyde. The most well established mechanism for maintaining low levels of free PLP is its dephosphorylation by phosphatases. A human PLP-specific phosphatase has been identified and characterized. However, very little is known about the phosphatase in other living organisms. In this study, a cDNA clone of putative PLP phosphatase was identified from B. mori and characterized. The cDNA encodes a polypeptide of 343 amino acid residues, and the recombinant enzyme purified from E. coli exhibited properties similar to that of human PLP phosphatase. B. mori has a single copy of the PLPP gene, which is located on 11th chromosome, spans a 5.7kb region and contains five exons and four introns. PLP phosphatase transcript was detected in every larva tissue except hemolymph, and was most highly represented in Malpighian tube. We further down-regulated the gene expression of the PLP phosphatase in 5th instar larvae with the RNA interference. However, no significant changes in the gene expression of PLP biosynthetic enzymes and composition of B 6 vitamers were detected as compared with the control. Copyright © 2017 Elsevier Inc. All rights reserved.

Paradox of the drinking-straw model of the butterfly proboscis.

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Tsai, Chen-Chih; Monaenkova, Daria; Beard, Charles E; Adler, Peter H; Kornev, Konstantin G

2014-06-15

Fluid-feeding Lepidoptera use an elongated proboscis, conventionally modeled as a drinking straw, to feed from pools and films of liquid. Using the monarch butterfly, Danaus plexippus (Linnaeus), we show that the inherent structural features of the lepidopteran proboscis contradict the basic assumptions of the drinking-straw model. By experimentally characterizing permeability and flow in the proboscis, we show that tapering of the food canal in the drinking region increases resistance, significantly hindering the flow of fluid. The calculated pressure differential required for a suction pump to support flow along the entire proboscis is greater than 1 atm (~101 kPa) when the butterfly feeds from a pool of liquid. We suggest that behavioral strategies employed by butterflies and moths can resolve this paradoxical pressure anomaly. Butterflies can alter the taper, the interlegular spacing and the terminal opening of the food canal, thereby controlling fluid entry and flow, by splaying the galeal tips apart, sliding the galeae along one another, pulsing hemolymph into each galeal lumen, and pressing the proboscis against a substrate. Thus, although physical construction of the proboscis limits its mechanical capabilities, its functionality can be modified and enhanced by behavioral strategies. © 2014. Published by The Company of Biologists Ltd.

Vitamin B12 and eicosanoids in insects

International Nuclear Information System (INIS)

Wakayama, J.E.

1985-01-01

Vitamin B 12 was not detected in the house fly, Musca domestica, which apparently cannot interconvert propionate and succinate. In contrast, the termite readily interconverts succinate and methylmalonate, and contains high amounts of vitamin B 12 . The intestinal bacteria were the major source of vitamin B 12 in the termite, Coptotermes formosanus. The presence of arachidonic acid (20:4) and eicosatrienoic acid (20:3,n-6) at low levels in adult male and female house flies was demonstrated by chemical ionization-gas chromatography-mass spectrometry. After injection, over 80% of 20:4 was rapidly incorporated into the phospholipid (PL) fraction. Over 80% of the sequestered 20:4 was in the 2-position of PLs. The 20:4 was injected into the insect or was included in the diet prior to administration of [ 3 H] 20:4; large amounts of radioactivity were recovered in the triacylglycerol and free fatty acid fractions. Arachidonic acid (20:4) injected into house flies was rapidly converted to prostaglandins, and was also catabolized rapidly. Radiolabeled 20:4 injected into the hemolymph was incorporated into the reproductive tissues of male insects. About 2.1% of the total radioactivity from [ 3 H] 20:4 injected into males just prior to mating was transferred to females during mating

ECOLOGY AND PARASITOLOGICAL ANALYSIS OF HORSE FLIES (DIPTERA: TABANIDAE IN ANTIOQUIA, COLOMBIA

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PARRA-HENAO GABRIEL

2008-07-01

Full Text Available During the months of June to September 2006, collections of tabanids (Diptera:Tabanidae and ticks were conducted in the Caucasia municipality, Antioquia,Colombia. Tabanids were caught on horses during daylight using hand nets and pots atthe ecotone zone between secondary forests and paddock habitats. Ticks were collecteddirectly from cattle by hand. The purpose of the study was to identify possible vectorsof bovine trypanosomosis, and register the diversity and abundance of tabanids inthe zone. The arthropods were brought to the laboratory for taxonomic determinationand protozooans searching in proboscis, midgut, and salivary glands of flies. Inthe case of ticks, protozoans were searched in hemolymph. One hundred and fortytabanids belonging to four genera and nine species were caught. Among the species,Lepiselaga crassipes was the most abundant (43.6%, with the highest abundancein July and a biting peak at 14:00 h. The highest diversity of tabanids was observedduring September. Three tabanids were found infected with flagellates morphologicallycompatible with Trypanosoma vivax. 315 ticks belonging to Boophilus microplusspecies were collected, all of them negative to flagellates. These results suggest T.vivax transmission by tabanids in the study area. However, the specific status ofthe parasites should be determined by molecular techniques and the transmissionmechanism should be established too by controlled studies

Quantitative assessment of viable Cryptosporidium parvum load in commercial oysters (Crassostrea virginica) in the Chesapeake Bay.

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Graczyk, Thaddeus K; Lewis, Earl J; Glass, Gregory; Dasilva, Alexandre J; Tamang, Leena; Girouard, Autumn S; Curriero, Frank C

2007-01-01

The epidemiological importance of increasing reports worldwide on Cryptosporidium contamination of oysters remains unknown in relation to foodborne cryptosporidiosis. Thirty market-size oysters (Crassostrea virginica), collected from each of 53 commercial harvesting sites in Chesapeake Bay, MD, were quantitatively tested in groups of six for Cryptosporidium sp. oocysts by immunofluorescent antibody (IFA). After IFA analysis, the samples were retrospectively retested for viable Cryptosporidium parvum oocysts by combined fluorescent in situ hybridization (FISH) and IFA. The mean cumulative numbers of Cryptosporidium sp. oocysts in six oysters (overall, 42.1+/-4.1) were significantly higher than in the numbers of viable C. parvum oocysts (overall, 28.0+/-2.9). Of 265 oyster groups, 221 (83.4%) contained viable C. parvum oocysts, and overall, from 10-32% (mean, 23%) of the total viable oocysts were identified in the hemolymph as distinct from gill washings. The amount of viable C. parvum oocysts was not related to oyster size or to the level of fecal coliforms at the sampling site. This study demonstrated that, although oysters are frequently contaminated with oocysts, the levels of viable oocysts may be too low to cause infection in healthy individuals. FISH assay for identification can be retrospectively applied to properly stored samples.

Uptake and retention of metallic nanoparticles in the Mediterranean mussel (Mytilus galloprovincialis)

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Hull, Matthew S.; Vikesland, Peter J. [Virginia Tech Department of Civil and Environmental Engineering, Blacksburg, VA (United States); Virginia Tech Institute for Critical Technology and Applied Science (ICTAS) (United States); Virginia Tech Center for Sustainable Nanotechnology (VT SuN) (United States); Center for the Environmental Implications of Nanotechnology (CEINT) (United States); Schultz, Irvin R., E-mail: ir_schultz@pnl.gov [Battelle Pacific Northwest National Laboratory (PNNL), Marine Sciences Laboratory, Sequim, WA (United States)

2013-09-15

Highlights: •We measured uptake of two types of nanomaterials in a marine mussel. •Uptake from water was rapid and complete in less than 24 h. •Only particles suspended in the water appeared to be absorbed. •Most absorbed nanomaterial was concentrated in the digestive gland. -- Abstract: We measured the uptake, distribution and elimination of two types of metallic nanoparticles (MetNPs) by the aquatic mussel Mytilus galloprovincialis in static seawater column exposures. Test MetNPs included polyethylene glycol (PEG) functionalized Fe{sub 3}O{sub 4} nanoparticles (PEG-FeOxNP) and PEG-functionalized cadmium-selenide quantum dots (PEG-Qdot). Exposure water was sampled at various times to assess MetNP clearance, and mussels were serially euthanized to assess uptake of MetNPs into the hemolymph, digestive gland, and remaining carcass. Results indicated that >90% of both types of MetNPs were taken up by mussels within 8 h of initial exposure. Nearly the entire retained dose of FeOxNPs and PEG-Qdots was deposited in the digestive gland. Our results provide important insights on the uptake and elimination kinetics of MetNPs in filter-feeding marine bivalves, and will be useful for subsequent development of toxicokinetic models to predict the kinetics of these processes.

Antiviral activity against human immunodeficiency virus-1 in vitro by myristoylated-peptide from Heliothis virescens

International Nuclear Information System (INIS)

Ourth, Donald D.

2004-01-01

An insect antiviral compound was purified from Heliothis virescens larval hemolymph by gel-filtration high pressure liquid chromatography (HPLC) and C-18 reverse-phase HPLC and its structure was determined by mass spectrometry. The antiviral compound is an N-myristoylated-peptide containing six amino acids with calculated molecular weight of 916 Da. The N-terminus contains the fatty acid myristoyl, and the C-terminus contains histidine with two methyl groups giving the histidine a permanent positive charge. The remainder of the compound is essentially non-polar. The structure of the compound corresponds with the 'myristate plus basic' motif expressed by certain viral proteins in their binding to the cytoplasmic side of the plasma membrane to initiate viral assembly and budding from a host cell. The insect antiviral compound may inhibit viral assembly and/or budding of viruses from host cells that could include the human immunodeficiency virus-1 (HIV-1) and herpes simplex virus-1 that use this motif for exit from a host cell. Using the formazan assay, the myristoylated-peptide was effective against HIV-1, with a nine times increase in the viability and protection in vitro of treated CEM-SS cells when compared with infected but untreated control cells

Phylogenetic diversity and functional characterization of the Manila clam microbiota: a culture-based approach.

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Leite, Laura; Jude-Lemeilleur, Florence; Raymond, Natalie; Henriques, Isabel; Garabetian, Frédéric; Alves, Artur

2017-09-01

According to the hologenome theory, the microbiota contributes to the fitness of the holobiont having an important role in its adaptation, survival, development, health, and evolution. Environmental stress also affects the microbiota and its capability to assist the holobiont in coping with stress factors. Here, we analyzed the diversity of cultivable bacteria associated with Manila clam tissues (mantle, gills, hemolymph) in two non-contaminated sites (Portugal and France) and one metal-contaminated site (Portugal). A total of 240 isolates were obtained. Representative isolates (n = 198) of the overall diversity were identified by 16S rDNA sequencing and subjected to functional characterization. Isolates affiliated with Proteobacteria, Actinobacteria, Firmicutes, and Bacteroidetes. Proteobacteria (mostly Pseudoalteromonadaceae and Vibrionaceae) were dominant in non-contaminated sites while Actinobacteria (mostly Microbacteriaceae) dominated in the metal-contaminated site. The main factor affecting the microbiota composition was contamination. No significant differences were observed between clam tissues and geographic regions. Several isolates tested positive for antibacterial activity, biofilm formation, protease, and siderophore production. The results show that the Manila clam harbors a diverse microbiota that may contribute to clam protection and overall fitness, as well as to its adaptation to stressful environments. In addition, the Manila clam microbiota is revealed as a promising source of novel probiotics with potential application in aquaculture.

Marine pollution risk in a coastal city: use of an eco-genotoxic tool as a stress indicator in mussels from the Eastern Aegean Sea.

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Kacar, Asli; Pazi, Idil; Gonul, Tolga; Kucuksezgin, Filiz

2016-08-01

Coastal areas, such as bays, estuaries, and harbors, are heavily polluted since these areas are the settlements to which toxic chemicals from industrial and domestic wastes are discharged. The genetic damage was evaluated using bioindicator mussel Mytilus galloprovincialis caused by toxic chemicals (metals and polycyclic aromatic hydrocarbons) in İzmir and Çandarlı Bays (the Eastern Aegean Sea) through comet assay. Three sampling sites from the two bays were selected and the study was conducted during the spring and autumn periods. The highest levels of DNA damage expressed as %Tail-DNA were observed in İzmir Bay (34.60 % Tail-DNA) in the spring. Analysis of the correlation between PAHs and metals in mussels and %T-DNA in the hemolymph and gill cells showed a statistically significant positive correlation between %T-DNA and ∑PAH, chromium (p llution level of the İzmir and Çandarlı Bays by using the DNA damage to the mussel, which can identify the effects of environmental pollutants at the cellular levels. These results confirm that comet assay can be used to determine the temporal and spatial differences of DNA damage, and as a suitable tool for the measurement of genotoxicity in regions with low pollutant concentrations.

Passive vectoring of entomopathogenic fungus Beauveria bassiana among the wax moth Galleria mellonella larvae by the ectoparasitoid Habrobracon hebetor females.

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Kryukov, Vadim Yu; Kryukova, Natalia A; Tyurin, Maksim V; Yaroslavtseva, Olga N; Glupov, Viktor V

2017-03-15

Females of the ectoparasitoid Habrobracon hebetor attack and envenomate numerous host individuals during oviposition. The vectoring of the entomopathogenic fungus Beauveria bassiana during the adhesion stage by ectoparasitoid females among the wax moth larvae Galleria mellonella was explored under laboratory conditions. Vectoring occurred both from infected parasitoids to wax moth larvae and from infected to healthy wax moth larvae by parasitoids. The efficacy of vectoring in both cases was dose dependent. Parasitoid females were unable to recognize infected larvae in a labyrinth test. In addition, the presence of H. hebetor females significantly (1.5-13 fold) increased the mycoses level in clusters of G. mellonella, with 40% of the larvae infected with fungal conidia. Envenomation by H. hebetor increased conidia germination on the cuticles of the wax moth larvae by 4.4 fold. An enhanced germination rate (2 fold) was registered in the n-hexane epicuticular extract of envenomated larvae compared to that of healthy larvae. Both envenomation and mycoses enhanced the phenoloxidase (PO) activity in the integument of G. mellonella and, in contrast, decreased the encapsulation rate in hemolymphs. We hypothesize that changes in the integument property and inhibition of cellular immunity provide the highest infection efficacy of entomopathogenic fungi with H. hebetor. © 2017 Institute of Zoology, Chinese Academy of Sciences.

White Shrimp Litopenaeus vannamei That Have Received Gracilaria tenuistipitata Extract Show Early Recovery of Immune Parameters after Ammonia Stressing.

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Chen, Yu-Yuan; Chen, Jiann-Chu; Lin, Yong-Chin; Yeh, Su-Tuen; Huang, Chien-Lun

2015-06-05

White shrimp Litopenaeus vannamei immersed in seawater (35‰) containing Gracilaria tenuistipitata extract (GTE) at 0 (control), 400, and 600 mg/L for 3 h were exposed to 5 mg/L ammonia-N (ammonia as nitrogen), and immune parameters including hyaline cells (HCs), granular cells (GCs, including semi-granular cells), total hemocyte count (THC), phenoloxidase (PO) activity, respiratory bursts (RBs), superoxide dismutase (SOD) activity, lysozyme activity, and hemolymph protein level were examined 24~120 h post-stress. The immune parameters of shrimp immersed in 600 mg/L GTE returned to original values earlier, at 96~120 h post-stress, whereas in control shrimp they did not. In another experiment, shrimp were immersed in seawater containing GTE at 0 and 600 mg/L for 3 h and examined for transcript levels of immune-related genes at 24 h post-stress. Transcript levels of lipopolysaccharide and β-1,3-glucan binding protein (LGBP), peroxinectin (PX), cytMnSOD, mtMnSOD, and HSP70 were up-regulated at 24 h post-stress in GTE receiving shrimp. We concluded that white shrimp immersed in seawater containing GTE exhibited a capability for maintaining homeostasis by regulating cellular and humoral immunity against ammonia stress as evidenced by up-regulated gene expression and earlier recovery of immune parameters.

Infection rate of Babesia spp. sporokinetes in engorged Boophilus microplus from an area of enzootic stability in the State of Minas Gerais, Brazil

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Quintão-Silva Maria G

2003-01-01

Full Text Available The infection rates of Babesia sporokinetes in engorged Boophilus microplus were evaluated during a 2-year period in a dairy farm located in an area of enzootic stability. Every 14 days engorged females were collected from calves and from adult animals. Ticks were incubated at 27 ± 0.5ºC and 80-90% relative humidity and Babesia infection rates were determined by microscopic examination of Giemsa-stained hemolymph smears. After 52 collections, 2105 ticks were obtained, from which 982 were collected from calves and 1123 from cows. The total Babesia infection rate was 10%, however the incidence was higher (p < 0.05 in ticks collected from calves (17.5% than in those collected from cows (3.6%. Females collected from cows showed the highest infection rates in January, March, and August, and absence of infection in April and May. Ticks feeding on calves were infected throughout the experimental period. The infection rates of engorged females collected from naturally infected calves that were artificially infested with Babesia-free-larvae of B. microplus gradually decreased until the calves were four months old. No differences were observed among infection rates of ticks collected from calves maintained under natural conditions.

Early gene Broad complex plays a key role in regulating the immune response triggered by ecdysone in the Malpighian tubules of Drosophila melanogaster.

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Verma, Puja; Tapadia, Madhu G

2015-08-01

In insects, humoral response to injury is accomplished by the production of antimicrobial peptides (AMPs) which are secreted in the hemolymph to eliminate the pathogen. Drosophila Malpighian tubules (MTs), however, are unique immune organs that show constitutive expression of AMPs even in unchallenged conditions and the onset of immune response is developmental stage dependent. Earlier reports have shown ecdysone positively regulates immune response after pathogenic challenge however, a robust response requires prior potentiation by the hormone. Here we provide evidence to show that MTs do not require prior potentiation with ecdysone hormone for expression of AMPs and they respond to ecdysone very fast even without immune challenge, although the different AMPs Diptericin, Cecropin, Attacin, Drosocin show differential expression in response to ecdysone. We show that early gene Broad complex (BR-C) could be regulating the IMD pathway by activating Relish and physically interacting with it to activate AMPs expression. BR-C depletion from Malpighian tubules renders the flies susceptible to infection. We also show that in MTs ecdysone signaling is transduced by EcR-B1 and B2. In the absence of ecdysone signaling the IMD pathway associated genes are down regulated and activation and translocation of transcription factor Relish is also affected. Copyright © 2015 Elsevier Ltd. All rights reserved.

Temporal and spatial resource use by female three-toed sloths and their young in an agricultural landscape in Costa Rica

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Oscar Ramirez

2011-12-01

Full Text Available The information on ecological behavior of wild sloths is very scarce. In this study we determined the home ranges and resources used by three adult female three-toed sloths (Bradypus variegatus and their four young in an agricultural matrix of cacao (Theobroma cacao, pasture, riparian forests and living fencerows in Costa Rica. Births occurred during November-December and the young became independent at five to seven months of age. Initially, mothers remained fixed in one or a few trees, but expanded their use of resources as young sloths became independent from them. Mothers initially guided the young to preferred food and cover resources, but they gradually left their young in small nucleus areas and colonized new areas for themselves. Home range sizes for young sloths (up to seven months of age varied between 0.04-0.6 hectares, while home range sizes for mothers varied from 0.04-25.0 hectares. During the maternal care period, 22 tree species were used, with the most common being Cecropia obtusifolia (30.9%, Coussapoa villosa (25.6%, Nectandra salicifolia (12.1%, Pterocarpus officinalis (5.8% and Samanea saman (5.4%. However, young sloths used only 20 tree species, with the most common being C. villosa (18.4%, S. saman (18.5 % and N. salicifolia (16.7%. The cacao agroforest was used only by mother sloths and never by their young following separation. However, in the riparian forest, both mother sloths and young used the tree species. A total of 28 tree species were used by the mother sloth; including the food species: C. obtusifolia, C. villosa, N. salicifolia and P. officinalis. However, the young used 18 trees species in this habitat with N. salicifolia and S. saman most commonly used, although they rested and fed during the day in C. obtusifolia, C. villosa and O. sinnuata. The cacao agroforest with adjacent riparian forests and fencerows provides an important habitat type that links the smaller secondary forests and other patches. Rev

The bandit, a new DNA transposon from a hookworm-possible horizontal genetic transfer between host and parasite.

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Thewarach Laha

2007-09-01

Full Text Available An enhanced understanding of the hookworm genome and its resident mobile genetic elements should facilitate understanding of the genome evolution, genome organization, possibly host-parasite co-evolution and horizontal gene transfer, and from a practical perspective, development of transposon-based transgenesis for hookworms and other parasitic nematodes.A novel mariner-like element (MLE was characterized from the genome of the dog hookworm, Ancylostoma caninum, and termed bandit. The consensus sequence of the bandit transposon was 1,285 base pairs (bp in length. The new transposon was flanked by perfect terminal inverted repeats of 32 nucleotides in length with a common target site duplication TA, and it encoded an open reading frame (ORF of 342 deduced amino acid residues. Phylogenetic comparisons confirmed that the ORF encoded a mariner-like transposase, which included conserved catalytic domains, and that the bandit transposon belonged to the cecropia subfamily of MLEs. The phylogenetic analysis also indicated that the Hsmar1 transposon from humans was the closest known relative of bandit, and that bandit and Hsmar1 constituted a clade discrete from the Tc1 subfamily of MLEs from the nematode Caenorhabditis elegans. Moreover, homology models based on the crystal structure of Mos1 from Drosophila mauritiana revealed closer identity in active site residues of the catalytic domain including Ser281, Lys289 and Asp293 between bandit and Hsmar1 than between Mos1 and either bandit or Hsmar1. The entire bandit ORF was amplified from genomic DNA and a fragment of the bandit ORF was amplified from RNA, indicating that this transposon is actively transcribed in hookworms.A mariner-like transposon termed bandit has colonized the genome of the hookworm A. caninum. Although MLEs exhibit a broad host range, and are identified in other nematodes, the closest phylogenetic relative of bandit is the Hsmar1 element of humans. This surprising finding suggests

Mid-Holocene to Present Climate Transition in Tropical South America

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Turcq, B.; Cordeiro, R.; Sifeddine, A.; Braconnot, P.; Dias, P. S.; Costa, R.; Jorgetti, T.

2008-12-01

The classical illustration of Holocene climate changes in tropical South America is the huge rising of Titicaca lake level from 4400 to 4000 cal BP. Because the Amazon basin is the source of Andean rainfalls we have explored Amazonian data of climate changes during the Holocene to better understand the cause of this abrupt transition. Amazonian data confirm the existence of mid-Holocene dryness: (1) lacustrine level studies show a lower precipitation/evaporation budget than present, with the lowest lake levels between 8500 and 6800 cal BP; (2) although the dominant Holocene vegetation has always been the rainforest in the heart of Amazonia, this forest expanded towards the northwestern and southwestern regions from 6800 to 1550 cal BP, moreover, pioneer elements of the rainforest developed during the mid-Holocene and the best example is those of Cecropia, between 9000 and 5000 cal BP. (3) soil d13C indicates a forest expansion over savannas areas in Roraima (north), Mato Grosso and Rondonia (southwest), during the Holocene. (4) the mid-Holocene (8000- 4000 cal BP) is characterized by repeated occurrences of forest fires, marked by the presence of charcoals in soils and lacustrine sediments. However these different records are not characterized by abrupt transitions at the end of the Middle Holocene in Amazonia. In the Andean records there is a clear north-south shift in the timing of the transition. Analysis of coupled Ocean Atmosphere Model simulations suggest that convection in Amazon basin is directly controlled by insolation leading to an almost linear response of local climate to the global forcing. Differently, in the eastern and south-western regions where the rain is brought by the South American Monsoon, the climate transition appears more abrupt. It may be because the involved climate mechanisms are more complex and depend on Ocean/Atmosphere/Vegetation coupled process (ITCZ position, ZCAS formation, etc.). Tectonic movements or threshold links to

Effect of salinity on the metabolism and osmoregulation of selected ontogenetic stages of an Amazon population of Macrobrachium amazonicum shrimp (Decapoda, Palaemonidae).

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Mazzarelli, C C M; Santos, M R; Amorim, R V; Augusto, A

2015-05-01

Probably as a function of their wide geographical distribution, the different population of Macrobrachium amazonicum shrimp may present distinct physiological, biochemical, reproductive, behavioral, and ecological patterns. These differences are so accentuated that the existence of allopatric speciation has been suggested, although initial studies indicate that the genetic variability of populations happen at an intraspecific level. Among the biological responses described for M. amazonicum populations, those regarding osmoregulation and metabolism play a key role for being related to the occupation of diverse habitats. To this effect, we investigated osmoregulation through the role of free amino acids in cell volume control and metabolism, through oxygen consumption in larvae (zoeae I, II, V and IX) and/or post-larvae of a M. amazonicum population from Amazon, kept in aquaculture fish hatcheries in the state of São Paulo. The results add information regarding the existence of distinct physiological responses among M. amazonicum populations and suggest that possible adjustments to metabolism and to the use of free amino acids as osmolytes of the regulation of the larvae and post-larvae cell volume depend on the appearance of structures responsible for hemolymph osmoregulation like, for example, the gills. In this respect, we verified that zoeae I do not alter their metabolism due to the exposition to fresh or brackish water, but they reduce intracellular concentration of free amino acids when exposed to fresh water, what may suggest the inexistence or inefficient performance of the structures responsible for volume regulation and hemolymph composition. On the other hand, in zoeae II and V exposed to fresh and brackish water, metabolism alterations were not followed by changes in free amino acids concentration. Thus it is possible, as the structures responsible for osmoregulation and ionic regulation become functional, that the role of free amino acids gets

Morphological and molecular description of Blastocrithidia cyrtomeni sp. nov. (Kinetoplastea: Trypanosomatidae associated with Cyrtomenus bergi Froeschner (Hemiptera: Cydnidae from Colombia

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Ana Milena Caicedo

2011-05-01

Full Text Available A new trypanosomatid species, Blastocrithidia cyrtomeni, is herein described using morphological and molecular data. It was found parasitising the alimentary tract of the insect host Cyrtomenus bergi, a polyphagous pest. The morphology of B. cyrtomeni was investigated using light and transmission microscopy and molecular phylogeny was inferred from the sequences of spliced leader RNA (SL rRNA - 5S rRNA gene repeats and the 18S small subunit (SSU rRNA gene. Epimastigotes of variable size with straphanger cysts adhering to the middle of the flagellum were observed in the intestinal tract, hemolymph and Malpighian tubules. Kinetoplasts were always observed anterior to the nucleus. The ultrastructure of longitudinal sections of epimastigotes showed the flagellum arising laterally from a relatively shallow flagellar pocket near the kinetoplast. SL RNA and 5S rRNA gene repeats were positive in all cases, producing a 0.8-kb band. The amplicons were 797-803 bp long with > 98.5% identity, indicating that they originated from the same organism. According to the sequence analysis of the SL-5S rRNA gene repeats and the 18S SSU rRNA gene, B. cyrtomeni is different from all other known species or isolates of Trypanosomatidae. Both analyses indicate that among known species, it is most closely related to Blastocrithidia triatomae.

Effects of ergothioneine from mushrooms (Flammulina velutipes) on melanosis and lipid oxidation of kuruma shrimp (Marsupenaeus japonicus).

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Encarnacion, Angel B; Fagutao, Fernand; Hirono, Ikuo; Ushio, Hideki; Ohshima, Toshiaki

2010-02-24

The antimelanosic and antioxidative properties of a hot water extract prepared from the fruiting body of the edible mushroom (Flammulina velutipes) were evaluated by dietary supplementation in Kuruma shrimp (Marsupenaeus japonicus) for possible aquaculture application. The extract contained ergothioneine (ERT) at a level of 2.05 mg/mL. A commercial standard of l-ergothioneine (l-ERT) and the mushroom extract showed inhibitory activity against mushroom polyphenoloxidase (PPO). Feeding of the extract had no adverse effects on the immune systems of the shrimp under the present experimental conditions. Supplementation of the extract in the diet significantly suppressed PPO activities in the hemolymphs of the shrimp. Expression of the prophenoloxidase (proPO) gene decreased in the hemocyte of the Kuruma shrimp fed with the mushroom extract. Consequently, development of melanosis was significantly suppressed in the supplement fed shrimp during ice storage. Lipid oxidation was also effectively controlled in the supplement fed group throughout the storage period. In vitro experiments showed that l-ERT effectively inhibited the activation of proPO in the hemocyte lysate supernatant (HLS). The transcript of the proPO gene in the hemocyte showed lower expression in the l-ERT-treated HLS. It was concluded that dietary supplementation of the mushroom extract in shrimp could be a promising approach to control post mortem development of melanosis and lipid oxidation in shrimp muscles.

Short communication. Impact of the amino acid proline on the cold hardiness of honey bee, Apis mellifera L.

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M. Mollaei

2013-07-01

Full Text Available Like many insects, honey bee can increase its cold tolerance through freeze avoidance, using antifreeze proteins (AFPs to lower its supercooling point (SCP. Proline is the most dominant amino acid in honey bee hemolymph, which can be obtained by the insect through feeding. In the current study the antifreeze activity of this amino acid was evaluated on worker honey bees, immediately before the start of cold season. The experiment was established on four treatments including three different concentrations of proline (1%, 3% and 4.35% diluted in 1:1 water sucrose syrup, and the syrup without proline (control. Newly emerged worker honey bees were fed on the mentioned diets for 2 weeks, under cage condition, and then 20 bees from each treatment (cage were selected randomly for determination of cold hardiness inside a cooling bath. Using a CHY data logger, equipped with a K100 sensor attached to the bee’s gaster, the SCP, the amount of released heat and the rate of this release as measures of insect cold hardiness were recorded. Proline significantly reduced honey bees’ SCP. The lowest point, -7.67±0.2646°C, was observed in the concentration of 1% proline. The amount of released heat and the rate of this release were not significantly different across the treatments.

Acclimatory responses of the Daphnia pulex proteome to environmental changes. I. Chronic exposure to hypoxia affects the oxygen transport system and carbohydrate metabolism

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Madlung Johannes

2009-04-01

Full Text Available Abstract Background Freshwater planktonic crustaceans of the genus Daphnia show a remarkable plasticity to cope with environmental changes in oxygen concentration and temperature. One of the key proteins of adaptive gene control in Daphnia pulex under hypoxia is hemoglobin (Hb, which increases in hemolymph concentration by an order of magnitude and shows an enhanced oxygen affinity due to changes in subunit composition. To explore the full spectrum of adaptive protein expression in response to low-oxygen conditions, two-dimensional gel electrophoresis and mass spectrometry were used to analyze the proteome composition of animals acclimated to normoxia (oxygen partial pressure [Po2]: 20 kPa and hypoxia (Po2: 3 kPa, respectively. Results The comparative proteome analysis showed an up-regulation of more than 50 protein spots under hypoxia. Identification of a major share of these spots revealed acclimatory changes for Hb, glycolytic enzymes (enolase, and enzymes involved in the degradation of storage and structural carbohydrates (e.g. cellubiohydrolase. Proteolytic enzymes remained constitutively expressed on a high level. Conclusion Acclimatory adjustments of the D. pulex proteome to hypoxia included a strong induction of Hb and carbohydrate-degrading enzymes. The scenario of adaptive protein expression under environmental hypoxia can be interpreted as a process to improve oxygen transport and carbohydrate provision for the maintenance of ATP production, even during short episodes of tissue hypoxia requiring support from anaerobic metabolism.

Induced resistance to infection of lobsters Homarus americanus by Aerococcus viridans (var.) homari, the bacterium causing gaffkemia.

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Stewart, James E; Arie, B; Marks, L J

2004-12-13

A vaccine composed of steam sterilized (autoclaved) cells of a virulent strain of Aerococcus viridans (var.) homari was effective in protecting lobsters Homarus americanus against gaffkemia. At 15 degrees C the heat-killed vaccines (HKV) at concentrations between 1 and 5 x 10(7) particles kg(-1) lobster body wt induced maximal protection in induction periods ranging from 7 to 11 d. Protection was substantial over the course of a 30 d post-induction trial period. Spring-caught lobsters (i.e. those more fully rehabilitated following ecdysis) gained more protection (LD50 = 1.9 x 10(4)) from the vaccination than did those caught in the late fall-early winter period (lobsters that were not yet fully recovered from ecdysis) (LD50 = 3.2 x 10(3)). The protection offered by the HK vaccine was comparable to that induced by a vaccine produced by incubating the pathogen with low concentrations (2 pg ml(-1)) of the antibiotic vancomycin. The bacterins produced by both methods exhibited similar new properties: (1) agglutination at low titres by lobster hemolymph serum, suggesting an impaired capsule layer, and (2) increased permeability to the large Alcian Blue molecule. With both vaccines, the protection may be a direct result of increased exposure to intact bacterial cell structures by the lobster defences, an exposure which otherwise would be prevented by an intact capsule.

Herbivory-induced glucose transporter gene expression in the brown planthopper, Nilaparvata lugens.

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Kikuta, Shingo; Nakamura, Yuki; Hattori, Makoto; Sato, Ryoichi; Kikawada, Takahiro; Noda, Hiroaki

2015-09-01

Nilaparvata lugens, the brown planthopper (BPH) feeds on rice phloem sap, containing high amounts of sucrose as a carbon source. Nutrients such as sugars in the digestive tract are incorporated into the body cavity via transporters with substrate selectivity. Eighteen sugar transporter genes of BPH (Nlst) were reported and three transporters have been functionally characterized. However, individual characteristics of NlST members associated with sugar transport remain poorly understood. Comparative gene expression analyses using oligo-microarray and quantitative RT-PCR revealed that the sugar transporter gene Nlst16 was markedly up-regulated during BPH feeding. Expression of Nlst16 was induced 2 h after BPH feeding on rice plants. Nlst16, mainly expressed in the midgut, appears to be involved in carbohydrate incorporation from the gut cavity into the hemolymph. Nlst1 (NlHT1), the most highly expressed sugar transporter gene in the midgut was not up-regulated during BPH feeding. The biochemical function of NlST16 was shown as facilitative glucose transport along gradients. Glucose uptake activity by NlST16 was higher than that of NlST1 in the Xenopus oocyte expression system. At least two NlST members are responsible for glucose uptake in the BPH midgut, suggesting that the midgut of BPH is equipped with various types of transporters having diversified manner for sugar uptake. Copyright © 2015 Elsevier Ltd. All rights reserved.

Natural selection on immune defense: A field experiment.

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Langeloh, Laura; Behrmann-Godel, Jasminca; Seppälä, Otto

2017-02-01

Predicting the evolution of phenotypic traits requires an understanding of natural selection on them. Despite its indispensability in the fight against parasites, selection on host immune defense has remained understudied. Theory predicts immune traits to be under stabilizing selection due to associated trade-offs with other fitness-related traits. Empirical studies, however, report mainly positive directional selection. This discrepancy could be caused by low phenotypic variation in the examined individuals and/or variation in host resource level that confounds trade-offs in empirical studies. In a field experiment where we maintained Lymnaea stagnalis snails individually in cages in a lake, we investigated phenotypic selection on two immune defense traits, phenoloxidase (PO)-like activity and antibacterial activity, in hemolymph. We used a diverse laboratory population and manipulated snail resource level by limiting their food supply. For six weeks, we followed immune activity, growth, and two fitness components, survival and fecundity of snails. We found that PO-like activity and growth were under stabilizing selection, while antibacterial activity was under positive directional selection. Selection on immune traits was mainly driven by variation in survival. The form of selection on immune defense apparently depends on the particular trait, possibly due to its importance for countering the present parasite community. © 2016 The Author(s). Evolution © 2016 The Society for the Study of Evolution.

Taste-independent detection of the caloric content of sugar in Drosophila.

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Dus, Monica; Min, SooHong; Keene, Alex C; Lee, Ga Young; Suh, Greg S B

2011-07-12

Feeding behavior is influenced primarily by two factors: nutritional needs and food palatability. However, the role of food deprivation and metabolic needs in the selection of appropriate food is poorly understood. Here, we show that the fruit fly, Drosophila melanogaster, selects calorie-rich foods following prolonged food deprivation in the absence of taste-receptor signaling. Flies mutant for the sugar receptors Gr5a and Gr64a cannot detect the taste of sugar, but still consumed sugar over plain agar after 15 h of starvation. Similarly, pox-neuro mutants that are insensitive to the taste of sugar preferentially consumed sugar over plain agar upon starvation. Moreover, when given a choice between metabolizable sugar (sucrose or D-glucose) and nonmetabolizable (zero-calorie) sugar (sucralose or L-glucose), starved Gr5a; Gr64a double mutants preferred metabolizable sugars. These findings suggest the existence of a taste-independent metabolic sensor that functions in food selection. The preference for calorie-rich food correlates with a decrease in the two main hemolymph sugars, trehalose and glucose, and in glycogen stores, indicating that this sensor is triggered when the internal energy sources are depleted. Thus, the need to replenish depleted energy stores during periods of starvation may be met through the activity of a taste-independent metabolic sensing pathway.

Molecular Cloning, Bioinformatic Analysis, and Expression of Bombyx mori Lebocin 5 Gene Related to Beauveria bassiana Infection.

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Lü, Dingding; Hou, Chengxiang; Qin, Guangxing; Gao, Kun; Chen, Tian; Guo, Xijie

2017-01-01

A full-length cDNA of lebocin 5 (BmLeb5) was first cloned from silkworm, Bombyx mori , by rapid amplification of cDNA ends. The BmLeb5 gene is 808 bp in length and the open reading frame encodes a 179-amino acid hydroxyproline-rich peptide. Bioinformatic analysis results showed that BmLeb5 owns an O-glycosylation site and four RXXR motifs as other lebocins. Sequence similarity and phylogenic analysis results indicated that lebocins form a multiple gene family in silkworm as cecropins. Quantitative real-time PCR analysis revealed that BmLeb5 was highest expressed in the fat body. In the silkworm larvae infected by Beauveria bassiana , the expression level of BmLeb5 was upregulated in the fat body and hemolymph which are the most important immune tissues in silkworm. The recombinant protein of BmLeb5 was for the first time successfully expressed with prokaryotic expression system and purified. There are no reports so far that the expression of lebocins could be induced by entomopathogenic fungus. Our study suggested that BmLeb5 might play an important role in the immune response of silkworm to defend B. bassiana infection. The results also provided helpful information for further studying the lebocin family functioned in antifungal immune response in the silkworm.

Insulin-like growth factor (IGF)-like peptide and 20-hydroxyecdysone regulate the growth and development of the male genital disk through different mechanisms in the silkmoth, Bombyx mori.

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Fujinaga, Daiki; Kohmura, Yusuke; Okamoto, Naoki; Kataoka, Hiroshi; Mizoguchi, Akira

2017-08-01

It is well established that ecdysteroids play pivotal roles in the regulation of insect molting and metamorphosis. However, the mechanisms by which ecdysteroids regulate the growth and development of adult organs after pupation are poorly understood. Recently, we have identified insulin-like growth factor (IGF)-like peptides (IGFLPs), which are secreted after pupation under the control of 20-hydroxyecdysone (20E). In the silkmoth, Bombyx mori, massive amounts of Bombyx-IGFLP (BIGFLP) are present in the hemolymph during pupal-adult development, suggesting its importance in the regulation of adult tissue growth. Thus, we hypothesized that the growth and development of adult tissues including imaginal disks are regulated by the combined effects of BIGFLP and 20E. In this study, we investigated the growth-promoting effects of BIGFLP and 20E using the male genital disks of B. mori cultured ex vivo, and further analyzed the cell signaling pathways mediating hormone actions. We demonstrate that 20E induces the elongation of genital disks, that both hormones stimulate protein synthesis in an additive manner, and that BIGFLP and 20E exert their effects through the insulin/IGF signaling pathway and mitogen-activated protein kinase pathway, respectively. These results show that the growth and development of the genital disk are coordinately regulated by both BIGFLP and 20E. Copyright © 2017 Elsevier Ltd. All rights reserved.

Rab proteins in the brain and corpus allatum of Bombyx mori.

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Uno, Tomohide; Furutani, Masayuki; Watanabe, Chihiro; Sakamoto, Katsuhiko; Uno, Yuichi; Kanamaru, Kengo; Yamagata, Hiroshi; Mizoguchi, Akira; Takeda, Makio

2016-07-01

In eukaryotic cells, Rab guanosine triphosphate-ases serve as key regulators of membrane-trafficking events, such as exocytosis and endocytosis. Rab3, Rab6, and Rab27 control the regulatory secretory pathway of neuropeptides and neurotransmitters. The cDNAs of Rab3, Rab6, and Rab27 from B. mori were inserted into a plasmid, transformed into Escherichia coli, and then subsequently purified. We then produced antibodies against Rab3, Rab6, and Rab27 of Bombyx mori in rabbits and rats for use in western immunoblotting and immunohistochemistry. Western immunoblotting of brain tissue revealed a single band at approximately 26 kDa. Immunohistochemistry results revealed that Rab3, Rab6, and Rab27 expression was restricted to neurons in the pars intercerebralis and dorsolateral protocerebrum of the brain. Rab3 and Rab6 co-localized with bombyxin, an insect neuropeptide. However, there was no Rab that co-localized with prothoracicotropic hormone. The corpus allatum secretes neuropeptides synthesized in the brain into the hemolymph. Results showed that Rab3 and Rab6 co-localized with bombyxin in the corpus allatum. These findings suggest that Rab3 and Rab6 are involved in neurosecretion in B. mori. This study is the first to report a possible relationship between Rab and neurosecretion in the insect corpus allatum.