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Sample records for thuringiensis vip3aa20 insecticidal

  1. Activity of vegetative insecticidal proteins Vip3Aa58 and Vip3Aa59 of Bacillus thuringiensis against lepidopteran pests.

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    Baranek, Jakub; Kaznowski, Adam; Konecka, Edyta; Naimov, Samir

    2015-09-01

    Vegetative insecticidal proteins (Vips) secreted by some isolates of Bacillus thuringiensis show activity against insects and are regarded as insecticides against pests. A number of B. thuringiensis strains harbouring vip3A genes were isolated from different sources and identified by using a PCR based approach. The isolates with the highest insecticidal activity were indicated in screening tests, and their vip genes were cloned and sequenced. The analysis revealed two polymorphic Vip protein forms, which were classified as Vip3Aa58 and Vip3Aa59. After expression of the vip genes, the proteins were isolated and characterized. The activity of both toxins was estimated against economically important lepidopteran pests of woodlands (Dendrolimus pini), orchards (Cydia pomonella) and field crops (Spodoptera exigua). Vip3Aa58 and Vip3Aa59 were highly toxic and their potency surpassed those of many Cry proteins used in commercial bioinsecticides. Vip3Aa59 revealed similar larvicidal activity as Vip3Aa58 against S. exigua and C. pomonella. Despite 98% similarity of amino acid sequences of both proteins, Vip3Aa59 was significantly more active against D. pini. Additionally the effect of proteolytic activation of Vip58Aa and Vip3Aa59 on toxicity of D. pini and S. exigua was studied. Both Vip3Aa proteins did not show any activity against Tenebrio molitor (Coleoptera) larvae. The results suggest that the Vip3Aa58 and Vip3Aa59 toxins might be useful for controlling populations of insect pests of crops and forests. Copyright © 2015 Elsevier Inc. All rights reserved.

  2. Selection and characterization of resistance to the Vip3Aa20 protein from Bacillus thuringiensis in Spodoptera frugiperda.

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    Bernardi, Oderlei; Bernardi, Daniel; Horikoshi, Renato J; Okuma, Daniela M; Miraldo, Leonardo L; Fatoretto, Julio; Medeiros, Fernanda Cl; Burd, Tony; Omoto, Celso

    2016-09-01

    Spodoptera frugiperda is one the main target pests of maize events expressing Vip3Aa20 protein from Bacillus thuringiensis (Bt) in Brazil. In this study, we selected a resistant strain of S. frugiperda on Bt maize expressing Vip3Aa20 protein and characterized the inheritance and fitness costs of the resistance. The resistance ratio of the Vip3Aa20-resistant strain of S. frugiperda was >3200-fold. Neonates of the Vip3Aa20-resistant strain were able to survive and emerge as fertile adults on Vip3Aa20 maize, while larvae from susceptible and heterozygous strains did not survive. The inheritance of Vip3Aa20 resistance was autosomal recessive and monogenic. Life history studies to investigate fitness cost revealed an 11% reduction in the survival rate until adult stage and a ∼50% lower reproductive rate of the Vip3Aa20-resistant strain compared with susceptible and heterozygous strains. This is the first characterization of S. frugiperda resistance to Vip3Aa protein. Our results provide useful information for resistance management programs designed to prevent or delay resistance evolution to Vip3Aa proteins in S. frugiperda. © 2016 Society of Chemical Industry. © 2016 Society of Chemical Industry.

  3. Susceptibility of Spodoptera frugiperda and S. exigua to Bacillus thuringiensis Vip3Aa insecticidal protein.

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    Chakroun, Maissa; Bel, Yolanda; Caccia, Silvia; Abdelkefi-Mesrati, Lobna; Escriche, Baltasar; Ferré, Juan

    2012-07-01

    The Vip3Aa protein is an insecticidal protein secreted by Bacillus thuringiensis during the vegetative stage of growth. The activity of this protein has been tested after different steps/protocols of purification using Spodoptera frugiperda as a control insect. The results showed that the Vip3Aa protoxin was stable and retained full toxicity after being subjected to common biochemical steps used in protein purification. Bioassays with the protoxin in S. frugiperda and S. exigua showed pronounced differences in LC(50) values when mortality was measured at 7 vs. 10d. At 7d most live larvae were arrested in their development. LC(50) values of "functional mortality" (dead larvae plus larvae remaining in the first instar), measured at 7d, were similar or even lower than the LC(50) values of mortality at 10d. This strong growth inhibition was not observed when testing the trypsin-activated protein (62 kDa) in either species. S. exigua was less susceptible than S. frugiperda to the protoxin form, with LC(50) values around 10-fold higher. However, both species were equally susceptible to the trypsin-activated form. Processing of Vip3Aa protoxin to the activated form was faster with S. frugiperda midgut juice than with S. exigua midgut juice. The results strongly suggest that the differences in the rate of activation of the Vip3Aa protoxin between both species are the basis for the differences in susceptibility towards the protoxin form. Copyright © 2012 Elsevier Inc. All rights reserved.

  4. Vip3Aa induces apoptosis in cultured Spodoptera frugiperda (Sf9) cells.

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    Jiang, Kun; Mei, Si-Qi; Wang, Ting-Ting; Pan, Jin-Hua; Chen, Yue-Hua; Cai, Jun

    2016-09-15

    The vegetative insecticidal proteins (Vip) secreted by many Bacillus thuringiensis strains during their vegetative growth stage are regarded as second generation insecticidal proteins, as they share no sequence or structural homology with known crystal insecticidal proteins (Cry) and have a broad insecticidal spectrum. Compared with insecticidal crystal proteins (ICPs), the insecticidal mechanisms of Vips have been little studied. Here we investigated the mechanism responsible for Vip3Aa toxicity in cultured insect cells. Using, flow cytometry analyzes, TUNEL staining and DNA fragmentation assays, we show that Vip3Aa can induce apoptosis in Spodoptera frugiperda (Sf9) cells and cause cells to arrest at the G2/M phase. We also show that Vip3Aa can disrupt mitochondrial membrane potential (ΔΨm), leading to the activation of Sf-caspase-1, suggesting that a mitochondrial mediated and caspase dependent pathway may be involved in Vip3Aa-induced apoptosis in Sf9 cells. Copyright © 2016 Elsevier Ltd. All rights reserved.

  5. In-silico determination of insecticidal potential of Vip3Aa-Cry1Ac fusion protein against Lepidopteran targets using molecular docking

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    Aftab eAhmad

    2015-12-01

    Full Text Available Study and research of Bt (Bacillus thuringiensis transgenic plants have opened new ways to combat insect pests. Over the decades, however, insect pests, especially the Lepidopteran, have developed tolerance against Bt delta-endotoxins. Such issues can be addressed through the development of novel toxins with greater toxicity and affinity against a broad range of insect receptors. In this computational study, functional domains of Bacillus thuringiensis crystal delta-endotoxin (Cry1Ac insecticidal protein and vegetative insecticidal protein (Vip3Aa have been fused to develop a broad-range Vip3Aa-Cry1Ac fusion protein. Cry1Ac and Vip3Aa are non-homologous insecticidal proteins possessing receptors against different targets within the midgut of insects. The insecticidal proteins were fused to broaden the insecticidal activity. Molecular docking analysis of the fusion protein against aminopeptidase-N (APN and cadherin receptors of five Lepidopteran insects (Agrotis ipsilon, Helicoverpa armigera, Pectinophora gossypiella, Spodoptera exigua and Spodoptera litura revealed that the Ser290, Ser293, Leu337, Thr340 and Arg437 residues of the fusion protein are involved in the interaction with insect receptors. The Helicoverpa armigera cadherin receptor, however, showed no interaction, which might be due to either loss or burial of interactive residues inside the fusion protein. These findings revealed that the Vip3Aa-Cry1Ac fusion protein has a strong affinity against Lepidopteran insect receptors and hence has a potential to be an efficient broad-range insecticidal protein.

  6. Bacillus thuringiensis Cry1Ia10 and Vip3Aa protein interactions and their toxicity in Spodoptera spp. (Lepidoptera).

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    Bergamasco, V B; Mendes, D R P; Fernandes, O A; Desidério, J A; Lemos, M V F

    2013-02-01

    The polyphagous pests belonging to the genus Spodoptera are considered to be among the most important causes of damage and are widely distributed throughout the Americas'. Due to the extensive use of genetically modified plants containing Bacillus thuringiensis genes that code for insecticidal proteins, resistant insects may arise. To prevent the development of resistance, pyramided plants, which express multiple insecticidal proteins that act through distinct mode of actions, can be used. This study analyzed the mechanisms of action for the proteins Cry1Ia10 and Vip3Aa on neonatal Spodoptera frugiperda, Spodoptera albula, Spodoptera eridania and Spodoptera cosmioides larvae. The interactions of these toxins with receptors on the intestinal epithelial membrane were also analyzed by binding biotinylated toxins to brush border membrane vesicles (BBMVs) from the intestines of these insects. A putative receptor of approximately 65 kDa was found by ligand blotting in all of these species. In vitro competition assays using biotinylated proteins have indicated that Vip3Aa and Cry1Ia10 do not compete for the same receptor for S. frugiperda, S. albula and S. cosmioides and that Vip3Aa was more efficient than Cry1Ia10 when tested individually, by bioassays. A synergistic effect of the toxins in S. frugiperda, S. albula and S. cosmioides was observed when they were combined. However, in S. eridania, Cry1Ia10 and Vip3Aa might compete for the same receptor and through bioassays Cry1Ia10 was more efficient than Vip3Aa and showed an antagonistic effect when the proteins were combined. These results suggest that using these genes to develop pyramided plants may not prove effective in preventing the development of resistance in S. eridiana. Copyright © 2012 Elsevier Inc. All rights reserved.

  7. Amélioration des activités insecticides des protéines Vip3 de Bacillus thuringiensis

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    Sameh SELLAMI

    2016-07-01

    Full Text Available Vip3 proteins were considered as a second generation of biopesticides. They are synthesized and secreted by Bacillus thuringiensis during the vegetative growth phase. Vip3 proteins, which are discovered in 1990, are of great interest for the control of Lepidopteran insects pests such as Agrotis ipsilon, Spodoptera littoralis, Spodoptera exigua and Spodoptera frugiperda. Many researches were conducted on the Vip3 proteins in order to enlarge their spectrum, improve their insecticidal activities and resolve the problems of resistance that appeared recently after the massive use of δ-endotoxins, considered as first generation of biopesticides. In this review, we tried to summarize research studies interested in the improvement of the insecticidal activities of Vip3 proteins.

  8. First report of detection of the putative receptor of Bacillus thuringiensis toxin Vip3Aa from black cutworm (Agrotis ipsilon

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    Gamal H. Osman

    2018-03-01

    Full Text Available Black cutworm (BCW Agrotis ipsilon, an economically important lepidopteran insect, has attracted a great attention. Bacillus thuringiensis (Bt is spore forming soil bacteria and is an excellent environment-friendly approach for the control of phytophagous and disease-transmitting insects. In fact, bio-pesticide formulations and insect resistant transgenic plants based on the bacterium Bt delta-endotoxin have attracted worldwide attention as a safer alternative to harmful chemical pesticides. The major objective of the current study was to understand the mechanism of interaction of Bt toxin with its receptor molecule(s. The investigation involved the isolation, identification, and characterization of a putative receptor – vip3Aa. In addition, the kinetics of vip toxin binding to its receptor molecule was also studied. The present data suggest that Vip3Aa toxin bound specifically with high affinity to a 48-kDa protein present at the brush border membrane vesicles (BBMV prepared from the midgut epithelial cells of BCW larvae. Keywords: Receptor, vip3Aa, Bacillus thuringiensis, BBMV

  9. In Vivo and In Vitro Binding of Vip3Aa to Spodoptera frugiperda Midgut and Characterization of Binding Sites by 125I Radiolabeling

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    Chakroun, Maissa

    2014-01-01

    Bacillus thuringiensis vegetative insecticidal proteins (Vip3A) have been recently introduced in important crops as a strategy to delay the emerging resistance to the existing Cry toxins. The mode of action of Vip3A proteins has been studied in Spodoptera frugiperda with the aim of characterizing their binding to the insect midgut. Immunofluorescence histological localization of Vip3Aa in the midgut of intoxicated larvae showed that Vip3Aa bound to the brush border membrane along the entire apical surface. The presence of fluorescence in the cytoplasm of epithelial cells seems to suggest internalization of Vip3Aa or a fragment of it. Successful radiolabeling and optimization of the binding protocol for the 125I-Vip3Aa to S. frugiperda brush border membrane vesicles (BBMV) allowed the determination of binding parameters of Vip3A proteins for the first time. Heterologous competition using Vip3Ad, Vip3Ae, and Vip3Af as competitor proteins showed that they share the same binding site with Vip3Aa. In contrast, when using Cry1Ab and Cry1Ac as competitors, no competitive binding was observed, which makes them appropriate candidates to be used in combination with Vip3A proteins in transgenic crops. PMID:25002420

  10. Insights into the Structure of the Vip3Aa Insecticidal Protein by Protease Digestion Analysis

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    Yolanda Bel

    2017-04-01

    Full Text Available Vip3 proteins are secretable proteins from Bacillus thuringiensis whose mode of action is still poorly understood. In this study, the activation process for Vip3 proteins was closely examined in order to better understand the Vip3Aa protein stability and to shed light on its structure. The Vip3Aa protoxin (of 89 kDa was treated with trypsin at concentrations from 1:100 to 120:100 (trypsin:Vip3A, w:w. If the action of trypsin was not properly neutralized, the results of SDS-PAGE analysis (as well as those with Agrotis ipsilon midgut juice equivocally indicated that the protoxin could be completely processed. However, when the proteolytic reaction was efficiently stopped, it was revealed that the protoxin was only cleaved at a primary cleavage site, regardless of the amount of trypsin used. The 66 kDa and the 19 kDa peptides generated by the proteases co-eluted after gel filtration chromatography, indicating that they remain together after cleavage. The 66 kDa fragment was found to be extremely resistant to proteases. The trypsin treatment of the protoxin in the presence of SDS revealed the presence of secondary cleavage sites at S-509, and presumably at T-466 and V-372, rendering C-terminal fragments of approximately 29, 32, and 42 kDa, respectively. The fact that the predicted secondary structure of the Vip3Aa protein shows a cluster of beta sheets in the C-terminal region of the protein might be the reason behind the higher stability to proteases compared to the rest of the protein, which is mainly composed of alpha helices.

  11. 40 CFR 174.501 - Bacillus thuringiensis Vip3Aa protein in corn and cotton; exemption from the requirement of a...

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    2010-07-01

    ... 40 Protection of Environment 23 2010-07-01 2010-07-01 false Bacillus thuringiensis Vip3Aa protein in corn and cotton; exemption from the requirement of a tolerance. 174.501 Section 174.501 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) PESTICIDE PROGRAMS PROCEDURES AND REQUIREMENTS FOR PLANT-INCORPORATED PROTECTANTS...

  12. The Vip3Ag4 Insecticidal Protoxin from Bacillus thuringiensis Adopts A Tetrameric Configuration That Is Maintained on Proteolysis

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    Leopoldo Palma

    2017-05-01

    Full Text Available The Vip3 proteins produced during vegetative growth by strains of the bacterium Bacillus thuringiensis show insecticidal activity against lepidopteran insects with a mechanism of action that may involve pore formation and apoptosis. These proteins are promising supplements to our arsenal of insecticidal proteins, but the molecular details of their activity are not understood. As a first step in the structural characterisation of these proteins, we have analysed their secondary structure and resolved the surface topology of a tetrameric complex of the Vip3Ag4 protein by transmission electron microscopy. Sites sensitive to proteolysis by trypsin are identified and the trypsin-cleaved protein appears to retain a similar structure as an octomeric complex comprising four copies each of the ~65 kDa and ~21 kDa products of proteolysis. This processed form of the toxin may represent the active toxin. The quality and monodispersity of the protein produced in this study make Vip3Ag4 a candidate for more detailed structural analysis using cryo-electron microscopy.

  13. Resistance Risk Assessment of Spodoptera frugiperda (Lepidoptera: Noctuidae) and Diatraea saccharalis (Lepidoptera: Crambidae) to Vip3Aa20 Insecticidal Protein Expressed in Corn.

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    Bernardi, Oderlei; Bernardi, Daniel; Amado, Douglas; Sousa, Renan S; Fatoretto, Julio; Medeiros, Fernanda C L; Conville, Jared; Burd, Tony; Omoto, Celso

    2015-12-01

    Transgenic Agrisure Viptera 3 corn that expresses Cry1Ab, Vip3Aa20, and EPSPS proteins and Agrisure Viptera expressing Vip3Aa20 are used for control of Spodoptera frugiperda (J.E. Smith) and Diatraea saccharalis (F.) in Brazil. To support a resistance management program, resistance risk assessment studies were conducted to characterize the dose expression of Vip3Aa20 protein and level of control against these species. The Vip3Aa20 expression in Agrisure Viptera 3 and Agrisure Viptera decreased from V6 to V10 stage of growth. However, Vip3Aa20 expression in Agrisure Viptera 3 at V6 and V10 stages was 13- and 16-fold greater than Cry1Ab, respectively. The Vip3Aa20 expression in lyophilized tissue of Agrisure Viptera 3 and Agrisure Viptera diluted 25-fold in an artificial diet caused complete larval mortality of S. frugiperda and D. saccharalis. In contrast, lyophilized tissue of Bt11 at the same dilution does not provide complete mortality of these species. Agrisure Viptera 3 and Agrisure Viptera also caused a high level of mortality against S. frugiperda and D. saccharalis. Moreover, 100% mortality was observed for S. frugiperda larvae (neonates through fifth-instar larvae) when fed in corn with the Vip trait technology. Viptera corn achieves a high level of control against S. frugiperda and D. saccharalis providing a high dose, which is an important determination to support the refuge strategy for an effective resistance management program. © 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.

  14. Caracterização do gene vip3A e toxicidade da proteína Vip3Aa50 à lagarta-do-cartucho e à lagarta-da-soja

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    Camila Soares Figueiredo

    2013-09-01

    Full Text Available O objetivo deste trabalho foi caracterizar o gene vip3A de Bacillus thuringiensis e verificar a toxicidade da proteína Vip3Aa50 a larvas da lagarta-do-cartucho (Spodoptera frugiperda e da lagarta-da-soja (Anticarsia gemmatalis. O gene vip3A foi amplificado por PCR, com iniciadores específicos, e gerou um fragmento de 2.370 pb. Esse fragmento foi clonado em vetor pGEM-T Easy e, em seguida, sequenciado, subclonado em vetor de expressão pET-28a (+ e inserido em células de Escherichia coli BL21 (DE3. A expressão da proteína Vip3Aa50 foi induzida por isopropil-β-D-1-tiogalactopiranosídeo (IPTG, visualizada em SDS-PAGE e detectada por "Western blot". Os ensaios de toxicidade revelaram alta atividade da proteína Vip3Aa50 contra as larvas neonatas da lagarta-da-soja e da lagarta-do-cartucho, com CL50 de 20,3 e 79,6 ng cm-2, respectivamente. O gene vip3Aa50 é um novo gene da classe vip3A.

  15. Transgenic cotton coexpressing Vip3A and Cry1Ac has a broad insecticidal spectrum against lepidopteran pests.

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    Chen, Wen-Bo; Lu, Guo-Qing; Cheng, Hong-Mei; Liu, Chen-Xi; Xiao, Yu-Tao; Xu, Chao; Shen, Zhi-Cheng; Wu, Kong-Ming

    2017-10-01

    Although farmers in China have grown transgenic Bt-Cry1Ac cotton to resist the major pest Helicoverpa armigera since 1997 with great success, many secondary lepidopteran pests that are tolerant to Cry1Ac are now reported to cause considerable economic damage. Vip3AcAa, a chimeric protein with the N-terminal part of Vip3Ac and the C-terminal part of Vip3Aa, has a broad insecticidal spectrum against lepidopteran pests and has no cross resistance to Cry1Ac. In the present study, we tested insecticidal activities of Vip3AcAa against Spodoptera litura, Spodoptera exigua, and Agrotis ipsilon, which are relatively tolerant to Cry1Ac proteins. The bioassay results showed that insecticidal activities of Vip3AcAa against these three pests are superior to Cry1Ac, and after an activation pretreatment, Vip3AcAa retained insecticidal activity against S. litura, S. exigua and A. ipsilon that was similar to the unprocessed protein. The putative receptor for this chimeric protein in the brush border membrane vesicle (BBMV) in the three pests was also identified using biotinylated Vip3AcAa toxin. To broaden Bt cotton activity against a wider spectrum of pests, we introduced the vip3AcAa and cry1Ac genes into cotton. Larval mortality rates for S. litura, A. ipsilon and S. exigua that had fed on this new cotton increased significantly compared with larvae fed on non-Bt cotton and Bt-Cry1Ac cotton in a laboratory experiment. These results suggested that the Vip3AcAa protein is an excellent option for a "pyramid" strategy for integrated pest management in China. Copyright © 2017 Elsevier Inc. All rights reserved.

  16. Comprehensive analysis of gene expression profiles of the beet armyworm Spodoptera exigua larvae challenged with Bacillus thuringiensis Vip3Aa toxin.

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    Yolanda Bel

    Full Text Available Host-pathogen interactions result in complex relationship, many aspects of which are not completely understood. Vip proteins, which are Bacillus thuringensis (Bt insecticidal toxins produced during the vegetative stage, are selectively effective against specific insect pests. This new group of Bt proteins represents an interesting alternative to the classical Bt Cry toxins because current data suggests that they do not share the same mode of action. We have designed and developed a genome-wide microarray for the beet armyworm Spodoptera exigua, a serious lepidopteran pest of many agricultural crops, and used it to better understand how lepidopteran larvae respond to the treatment with the insecticidal protein Vip3Aa. With this approach, the goal of our study was to evaluate the changes in gene expression levels caused by treatment with sublethal doses of Vip3Aa (causing 99% growth inhibition at 8 and 24 h after feeding. Results indicated that the toxin provoked a wide transcriptional response, with 19% of the microarray unigenes responding significantly to treatment. The number of up- and down-regulated unigenes was very similar. The number of genes whose expression was regulated at 8 h was similar to the number of genes whose expression was regulated after 24 h of treatment. The up-regulated sequences were enriched for genes involved in innate immune response and in pathogen response such as antimicrobial peptides (AMPs and repat genes. The down-regulated sequences were mainly unigenes with homology to genes involved in metabolism. Genes related to the mode of action of Bt Cry proteins were found, in general, to be slightly overexpressed. The present study is the first genome-wide analysis of the response of lepidopteran insects to Vip3Aa intoxication. An insight into the molecular mechanisms and components related to Vip intoxication will allow designing of more effective management strategies for pest control.

  17. Impact of Spodoptera frugiperda neonate pretreatment conditions on Vip3Aa19 insecticidal protein activity and laboratory bioassay variation.

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    Da Silva, Karen F; Spencer, Terence A; Camargo Gil, Carolina; Siegfried, Blair D; Walters, Frederick S

    2016-04-01

    Variation in response to insecticidal proteins is common upon repetition of insect bioassays. Understanding this variation is a prerequisite to detecting biologically important differences. We tracked neonate Spodoptera frugiperda (J.E. Smith) susceptibility to Vip3Aa19 over 17 generations using standardized bioassay methods. Five larval pretreatment conditions and one bioassay condition were tested to determine whether susceptibility was affected. These included: storage time; prefeeding; storage at reduced temperature; storage at reduced humidity; colony introgression of field-collected individuals. Extremes of photoperiod during the bioassay itself were also examined. LC50 values for two strains of S. frugiperda varied 6.6-fold or 8.8-fold over 17 generations. Storage time and humidity had no impact on Vip3Aa19 susceptibility, whereas prefeeding significantly reduced subsequent mortality (by 27%). Storage at reduced temperature increased mortality for one colony (from 45.6 to 73.0%) but not for the other. Introgression of field-collected individuals affected susceptibility at the first generation but not for subsequent generations. A 24 h bioassay photophase significantly reduced susceptibility (by 26%) for both colonies. Certain pretreatment and bioassay conditions were identified that can affect S. frugiperda Vip3Aa19 susceptibility, but innate larval heterogeneity was also present. Our observations should help to increase the consistency of insecticidal protein bioassay results. © 2015 Syngenta Crop Protection, LLC. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

  18. Synergism and Antagonism between Bacillus thuringiensis Vip3A and Cry1 Proteins in Heliothis virescens, Diatraea saccharalis and Spodoptera frugiperda

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    Lemes, Ana Rita Nunes; Davolos, Camila Chiaradia; Legori, Paula Cristina Brunini Crialesi; Fernandes, Odair Aparecido; Ferré, Juan; Lemos, Manoel Victor Franco; Desiderio, Janete Apparecida

    2014-01-01

    Second generation Bt crops (insect resistant crops carrying Bacillus thuringiensis genes) combine more than one gene that codes for insecticidal proteins in the same plant to provide better control of agricultural pests. Some of the new combinations involve co-expression of cry and vip genes. Because Cry and Vip proteins have different midgut targets and possibly different mechanisms of toxicity, it is important to evaluate possible synergistic or antagonistic interactions between these two classes of toxins. Three members of the Cry1 class of proteins and three from the Vip3A class were tested against Heliothis virescens for possible interactions. At the level of LC50, Cry1Ac was the most active protein, whereas the rest of proteins tested were similarly active. However, at the level of LC90, Cry1Aa and Cry1Ca were the least active proteins, and Cry1Ac and Vip3A proteins were not significantly different. Under the experimental conditions used in this study, we found an antagonistic effect of Cry1Ca with the three Vip3A proteins. The interaction between Cry1Ca and Vip3Aa was also tested on two other species of Lepidoptera. Whereas antagonism was observed in Spodoptera frugiperda, synergism was found in Diatraea saccharalis. In all cases, the interaction between Vip3A and Cry1 proteins was more evident at the LC90 level than at the LC50 level. The fact that the same combination of proteins may result in a synergistic or an antagonistic interaction may be an indication that there are different types of interactions within the host, depending on the insect species tested. PMID:25275646

  19. A Novel Tenebrio molitor Cadherin Is a Functional Receptor for Bacillus thuringiensis Cry3Aa Toxin*

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    Fabrick, Jeff; Oppert, Cris; Lorenzen, Marcé D.; Morris, Kaley; Oppert, Brenda; Jurat-Fuentes, Juan Luis

    2009-01-01

    Cry toxins produced by the bacterium Bacillus thuringiensis are effective biological insecticides. Cadherin-like proteins have been reported as functional Cry1A toxin receptors in Lepidoptera. Here we present data that demonstrate that a coleopteran cadherin is a functional Cry3Aa toxin receptor. The Cry3Aa receptor cadherin was cloned from Tenebrio molitor larval midgut mRNA, and the predicted protein, TmCad1, has domain structure and a putative toxin binding region similar to those in lepidopteran cadherin B. thuringiensis receptors. A peptide containing the putative toxin binding region from TmCad1 bound specifically to Cry3Aa and promoted the formation of Cry3Aa toxin oligomers, proposed to be mediators of toxicity in lepidopterans. Injection of TmCad1-specific double-stranded RNA into T. molitor larvae resulted in knockdown of the TmCad1 transcript and conferred resistance to Cry3Aa toxicity. These data demonstrate the functional role of TmCad1 as a Cry3Aa receptor in T. molitor and reveal similarities between the mode of action of Cry toxins in Lepidoptera and Coleoptera. PMID:19416969

  20. A Novel Tenebrio molitor Cadherin Is a Functional Receptor for Bacillus thuringiensis Cry3Aa Toxin*

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    Fabrick, Jeff; Oppert, Cris; Lorenzen, Marcé D.; Morris, Kaley; Oppert, Brenda; Jurat-Fuentes, Juan Luis

    2009-01-01

    Cry toxins produced by the bacterium Bacillus thuringiensis are effective biological insecticides. Cadherin-like proteins have been reported as functional Cry1A toxin receptors in Lepidoptera. Here we present data that demonstrate that a coleopteran cadherin is a functional Cry3Aa toxin receptor. The Cry3Aa receptor cadherin was cloned from Tenebrio molitor larval midgut mRNA, and the predicted protein, TmCad1, has domain structure and a putative toxin binding region similar to those in lepid...

  1. Sub-lethal effects of Vip3A toxin on survival, development and fecundity of Heliothis virescens and Plutella xylostella.

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    Gulzar, Asim; Wright, Denis J

    2015-11-01

    The assessment of sub-lethal effects is important to interpret the overall insecticide efficacy in controlling insect pest populations. In addition to the lethal effect, sub-lethal effects may also occur in exposed insects. Vegetative insecticidal proteins (Vips) have shown a broad spectrum of insecticidal activity against many insect pest species. In this study the sub-lethal effects of the Bacillus thuringiensis vegetative insecticidal toxin Vip3A on the development and reproduction of Heliothis virescens F. and Plutella xylostella L. were evaluated in the laboratory. The results indicated that the sub-lethal concentration of Vip3A increased the duration of the larval and pupal stages as compared with the control treatment for both species. The percent pupation and percent adult emergence were significantly lower for Vip3A-treated insects. The proportion of pairs that produced eggs and the longevity of adults were not significantly different between treatments. H. virescens and P. xylostella treated with Vip3A showed an 11 and 17 % decrease in their intrinsic rate of increase (rm) respectively compared with untreated insects. The results from this study will be helpful to develop the strategy to incorporate Vip 3A containing crops in an integrated pest management programme.

  2. Bacillus thuringiensis monogenic strains: screening and interactions with insecticides used against rice pests

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    Pinto, Laura M.N.; Dörr, Natália C.; Ribeiro, Ana Paula A.; de Salles, Silvia M.; de Oliveira, Jaime V.; Menezes, Valmir G.; Fiuza, Lidia M.

    2012-01-01

    The screening of Bacillus thuringiensis (Bt) Cry proteins with high potential to control insect pests has been the goal of numerous research groups. In this study, we evaluated six monogenic Bt strains (Bt dendrolimus HD-37, Bt kurstaki HD-1, Bt kurstaki HD-73, Bt thuringiensis 4412, Bt kurstaki NRD-12 and Bt entomocidus 60.5, which codify the cry1Aa, cry1Ab, cry1Ac, cry1Ba, cry1C, cry2A genes respectively) as potential insecticides for the most important insect pests of irrigated rice: Spodoptera frugiperda, Diatraea saccharalis, Oryzophagus oryzae, Oebalus poecilus and Tibraca limbativentris. We also analyzed their compatibility with chemical insecticides (thiamethoxam, labdacyhalothrin, malathion and fipronil), which are extensively used in rice crops. The bioassay results showed that Bt thuringiensis 4412 and Bt entomocidus 60.5 were the most toxic for the lepidopterans, with a 93% and 82% mortality rate for S. frugiperda and D. saccharalis, respectively. For O. oryzae, the Bt kurstaki NRD-12 (64%) and Bt dendrolimus HD-37 (62%) strains were the most toxic. The Bt dendrolimus HD-37 strain also caused high mortality (82%) to O. poecilus, however the strains assessed to T. limbativentris caused a maximum rate of 5%. The assays for the Bt strains interaction with insecticides revealed the compatibility of the six strains with the four insecticides tested. The results from this study showed the high potential of cry1Aa and cry1Ba genes for genetic engineering of rice plants or the strains to biopesticide formulations. PMID:24031872

  3. BACILLUS THURINGIENSIS ELASTASES WITH INSECTICIDE ACTIVITY

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    E. V. Matseliukh

    2015-10-01

    Full Text Available The purpose of the research was a screening of proteases with elastase activity among Bacillus thuringiensis strains, their isolation, partially purification, study of physicochemical properties and insecticide activity in relation to the larvae of the Colorado beetle. The objects of the investigation were 18 strains of B. thuringiensis, isolated from different sources: sea water, dry biological product "Bitoksibatsillin" and also from natural populations of Colorado beetles of the Crimea, Kherson, Odesa, Mykolaiv and Zaporizhiia regions of Ukraine. Purification of enzymes with elastase activity isolated from above mentioned strains was performed by gel-chromatography and insecticide activity was studied on the 3–4 larvae instar of Colorado beetle. The ability of a number of B. thuringiensis strains to synthesize the proteases with elastase activity has been established. The most active were enzymes obtained from strains IMV B-7465, IMV B-7324 isolated from sea water, and strains 9, 902, Bt-H and 0-239 isolated from Colorado beetles. The study of the physicochemical properties of the partially purified proteases of these strains showed that they belonged to enzymes of the serine type. Peptidases of a number of B. thuringiensis strains (IMV B-7324, IMV B-7465, 902, 0-239, 9 are metal-dependent enzymes. Optimal conditions of action of all tested enzymes are the neutral and alkaline рН values and the temperatures of 30–40 °С. The studies of influence of the complex enzyme preparations and partially purified ones of B. thuringiensis strains on the larvae instar of Colorado beetles indicated that enzymes with elastase activity could be responsible for insecticide action of the tested strains.

  4. Detection and characterization of recombinant DNA expressing vip3A-type insecticidal gene in GMOs--standard single, multiplex and construct-specific PCR assays.

    Science.gov (United States)

    Singh, Chandra K; Ojha, Abhishek; Bhatanagar, Raj K; Kachru, Devendra N

    2008-01-01

    Vegetative insecticidal protein (Vip), a unique class of insecticidal protein, is now part of transgenic plants for conferring resistance against lepidopteron pests. In order to address the imminent regulatory need for detection and labeling of vip3A carrying genetically modified (GM) products, we have developed a standard single PCR and a multiplex PCR assay. As far as we are aware, this is the first report on PCR-based detection of a vip3A-type gene (vip-s) in transgenic cotton and tobacco. Our assay involves amplification of a 284-bp region of the vip-s gene. This assay can possibly detect as many as 20 natural wild-type isolates bearing a vip3A-like gene and two synthetic genes of vip3A in transgenic plants. The limit of detection as established by our assay for GM trait (vip-s) is 0.1%. Spiking with nontarget DNA originating from diverse plant sources had no inhibitory effect on vip-s detection. Since autoclaving of vip-s bearing GM leaf samples showed no deterioration/interference in detection efficacy, the assay seems to be suitable for processed food products as well. The vip-s amplicon identity was reconfirmed by restriction endonuclease assay. The primer set for vip-s was equally effective in a multiplex PCR assay format (duplex, triplex and quadruplex), used in conjunction with the primer sets for the npt-II selectable marker gene, Cauliflower mosaic virus 35S promoter and nopaline synthetase terminator, enabling concurrent detection of the transgene, regulatory sequences and marker gene. Further, the entire transgene construct was amplified using the forward primer of the promoter and the reverse primer of the terminator. The resultant amplicon served as a template for nested PCR to confirm the construct integrity. The method is suitable for screening any vip3A-carrying GM plant and food. The availability of a reliable PCR assay method prior to commercial release of vip3A-based transgenic crops and food would facilitate rapid and efficient regulatory

  5. Managing fall armyworm, Spodoptera frugiperda (Lepidoptera: Noctuidae), with Bt maize and insecticides in southern Brazil.

    Science.gov (United States)

    Burtet, Leonardo M; Bernardi, Oderlei; Melo, Adriano A; Pes, Maiquel P; Strahl, Thiago T; Guedes, Jerson Vc

    2017-12-01

    Maize plants expressing insecticidal proteins of Bacillus thuringiensis are valuable options for managing fall armyworm (FAW), Spodoptera frugiperda, in Brazil. However, control failures were reported, and therefore insecticides have been used to control this species. Based on these, we evaluated the use of Bt maize and its integration with insecticides against FAW in southern Brazil. Early-planted Agrisure TL, Herculex, Optimum Intrasect and non-Bt maize plants were severely damaged by FAW and required up to three insecticidal sprays. In contrast, YieldGard VT Pro, YieldGard VT Pro 3, PowerCore, Agrisure Viptera and Agrisure Viptera 3 showed little damage and did not require insecticides. Late-planted Bt maize plants showed significant damage by FAW and required up to four sprays, with the exceptions of Agrisure Viptera and Agrisure Viptera 3. Exalt (first and second sprays); Lannate + Premio (first spray) and Avatar (second spray); and Karate + Match (first spray) and Ampligo (second spray) were the most effective insecticides against FAW larvae in Bt and non-Bt maize. Maize plants expressing Cry proteins exhibited FAW control failures in southern Brazil, necessitating insecticidal sprays. In contrast, Bt maize containing the Vip3Aa20 protein remained effective against FAW. However, regardless of the insecticide used against FAW surviving on Bt maize, grain yields were similar. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

  6. Interação de proteínas Cry1 e Vip3A de Bacillus thuringiensis para controle de lepidópteros-praga

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    Paula Cristina Brunini Crialesi-Legori

    2014-02-01

    Full Text Available O objetivo deste trabalho foi avaliar a suscetibilidade das lagartas Anticarsia gemmatalis (Lepidoptera: Erebidae e Chrysodeixis includens (Lepidoptera: Noctuidae às proteínas Cry1 e Vip3A, bem como determinar se há a interação entre essas proteínas no controle das duas espécies. Bioensaios com as proteínas isoladas e em combinações foram realizados, e as concentrações letais CL50 e CL90 foram estimadas para cada condição. As proteínas Cry1Aa, Cry1Ac e Vip3Af foram as mais efetivas no controle de A. gemmatalis, enquanto Cry1Ac, Vip3Aa e Vip3Af foram mais efetivas no de C. includens. As proteínas Cry1Ac e Cry1Ca causaram maior inibição do desenvolvimento das larvas sobreviventes à CL50, em ambas as espécies. Combinações entre Vip3A e Cry1 apresentam efeito sinérgico no controle das espécies e a combinação Vip3Aa+Cry1Ea destaca-se no controle de A. gemmatalis e C. includens. Essas proteínas combinadas são promissoras na construção de plantas piramidadas, para o controle simultâneo das pragas.

  7. Resistance: a threat to the insecticidal crystal proteins of Bacillus thuringiensis

    Science.gov (United States)

    Leah S. Bauer

    1995-01-01

    Insecticidal crystal proteins (also known as d-endotoxins) synthesized by the bacterium Bacillus thuringiensis Berliner (Bt) are the active ingredient of various environmentally friendly insecticides that are 1) highly compatible with natural enemies and other nontarget organisms due to narrow host specificity, 2) harmless to vertebrates, 3) biodegradable in the...

  8. Environmental Distribution and Diversity of Insecticidal Proteins of Bacillus thuringiensis Berliner

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    Xavier, R.

    2007-01-01

    Full Text Available Bacillus thuringiensis Berliner based biopesticides have been successfully used world over for the control of agricultural pests and vectors of human diseases. Currently there are more than 200 B. thuringiensis strains with differing insecticidal activities are available as biocontrol agents and for developing transgenic plants. However, two major disadvantages are the development of insect resistance and high target specificity (narrow host range. Globally there is a continuous search for new B. thuringiensis strains with novel insecticidal activities. The present study aims to study the environmental distribution of B. thuringiensis and their toxic potential against insect pests. Soil and grain samples were collected from different environments and were processed by a modified acetate selection method. Initially B. thuringiensis isolates were screened on the basis of colony morphology and phase contrast microscopy for the presence of parasporal crystal inclusions. The population dynamics showed that B. thuringiensis is abundant in sericulture environment compared to other niches. Relative abundance of B. thuringiensis strains in sericulture environment shows the persistent association of B. thuringiensis with Bombyx mori (silk worm as insect pathogen. The protein profiles of the selected strains were studied by SDS-PAGE. The protein profiles of majority of B. thuringiensis isolates from grain storage facilities predominantly showing the 130 kDa and 68 kDa proteins, which is characteristics of lepidopteran active B. thuringiensis. However, one isolate BTRX-4 has 80-85 kDa protein, which is novel in that, this strain also exhibits antilepidopteran activity, which is normally presented by B. thuringiensis strains having 130 kDa and 68 kDa proteins. The protein profile of B. thuringiensis isolates from sericulture environment shows two different protein profiles. B. thuringiensis isolates BTRX-16 to BTRX-22 predominantly show 130 kDa protein

  9. Interactions of transgenic Bacillus thuringiensis insecticidal crops with spiders (Araneae)

    Science.gov (United States)

    Genetically modified crops expressing insecticidal proteins from Bacillus thuringiensis (Bt) have dramatically increased in acreage since their introduction in the mid-1990’s. Although the insecticidal mechanisms of Bt target specific pests, concerns persist regarding direct and indirect effects on...

  10. Adsorption of Insecticidal Crystal Protein Cry11Aa onto Nano-Mg(OH)2: Effects on Bioactivity and Anti-Ultraviolet Ability.

    Science.gov (United States)

    Pan, Xiaohong; Xu, Zhangyan; Li, Lan; Shao, Enshi; Chen, Saili; Huang, Tengzhou; Chen, Zhi; Rao, Wenhua; Huang, Tianpei; Zhang, Lingling; Wu, Songqing; Guan, Xiong

    2017-11-01

    The traditional Bacillus thuringiensis (Bt) formulations for field applications are not resistant to harsh environmental conditions. Hence, the active ingredients of the Bt bioinsecticides could degrade quickly and has low anti-ultraviolet ability in the field, which significantly limits its practical application. In the present study, we developed an efficient and stable delivery system for Bt Cry11Aa toxins. We coated Cry11Aa proteins with Mg(OH) 2 nanoparticles (MHNPs), and then assessed the effects of MHNPs on bioactivity and anti-ultraviolet ability of the Cry11Aa proteins. Our results indicated that MHNPs, like "coating clothes", could effectively protect the Cry protein and enhance the insecticidal bioactivity after UV radiation (the degradation rate was decreased from 64.29% to 16.67%). In addtion, MHNPs could improve the proteolysis of Cry11Aa in the midgut and aggravate the damage of the Cry protein to the gut epithelial cells, leading to increased insecticidal activity against Culex quinquefasciatus. Our results revealed that MHNPs, as an excellent nanocarrier, could substantially improve the insecticidal bioactivity and anti-ultraviolet ability of Cry11Aa.

  11. Bacillus thuringiensis Cry3Aa toxin increases the susceptibility of Crioceris quatuordecimpunctata to Beauveria bassiana infection

    Science.gov (United States)

    The spotted asparagus beetle, Crioceris quatuordecimpunctata (Coleoptera: Chrysomelidae), is one of the most devastating pests of asparagus in China and elsewhere. In this study, we investigated the interaction of Bacillus thuringiensis (Bt) Cry3Aa toxin and the entomopathogenic fungus Beauveria bas...

  12. Expression of Bacillus thuringiensis serovar. israelensis toxins in Asticcacaulis excentricus to control dipteran larvae of vectors of diseases

    Directory of Open Access Journals (Sweden)

    Óscar Enrique Guevara

    2004-01-01

    Full Text Available Bacillus thuringiensis cry genes encode for a diverse group of crystal-forming proteins that exhibit insecticidal activity towards dipteran, lepidopteran and coleopteran larvae. The effectiveness of insecticides based on mosquito larvicidal B. thuringiensis strains can be enhanced by using aquatic prosthecated bacteria as alternative hosts, since they do not sink, cytoplasmic located toxins are protected f rom UV radiation and, most importantly, mosquito larvae feed on them. An Asticcacaulis excentricus reference strain was transformed with the cry1 1Aa gene from Bacillus thuringiensis serovar. israelensis. Western blot and electrophoresis were used to test recombinant protein expression; Western blot revealed a 72 kDa protein corresponding to B. thuringiensis serovar. israelensis Cry1 1 Aa. These aquatic bacte­rias toxicity achieved 50% mortality at 23 ng/mL concentration in f irst instar Culex quinquefasciatus larvae. Other bioassays indicated that recombinant A. excentricus is toxic against Aedes aegyptiand Anopheles albimanus first instar larvae. Buoyancy tests demonstrated the advantage of A. excentricus over B. thuringiensis. Key words: Asticcacaulis excentricus, Bacillus thuringiensis, prosthecated bacteria, dengue, malaria.

  13. Expresión de la toxina Cry11Aa de Bacillus thuringiensis serovar. israelensis en Asticcacaulis excentricus, para el control de larvas acuáticas de dípteros de la familia Culicidae, vectores de enfermedades Expression of Bacillus thuringiensis serovar. israelensis toxins in Asticcacaulis excentricus to control dipteran larvae of vectors of diseases

    Directory of Open Access Journals (Sweden)

    Orduz Sergio

    2004-07-01

    Full Text Available Los genes cry de Bacillus thuringiensis codifican para un diverso grupo de proteínas formadoras de cristales que exhiben actividad insecticida contra larvas de dípteros, lepidópteros y coleópteros, entre otros. La efectividad de los insecticidas basados en formulaciones de proteínas de B. thuringiensis puede ser mejorada usando bacterias prostecadas acuáticas como portadoras alternativas de los genes cry, ya que no se sedimentan rápidamente; las proteínas expresadas en el citoplasma están protegidas de los rayos ultra violeta y, lo más importante, las larvas de los mosquitos se alimentan de ellas. Una cepa de referencia de Asticcacaulis excentricus fue transformada con el plásmido pSOD3, el cual contiene el gen que codifica para la proteína Cry1 1 Aa de B. thuringiensis serovar. israelensis. La expresión de la proteína recombinante fue evaluada por electroforesis de proteínas y por Western blot. El Western blot revelado con un anticuerpo policlonal anti-Cry1 1 Aa mostró una banda de 72 kDa correspondiente a la proteína Cry 11 Aa. La toxicidad de las cepas de A. excentricus transformadas fue evaluada en bioensayos con larvas de primer estadío del mosquito Culex quinquefasciatus. Se alcanzó un promedio de mortalidad del 50% de las larvas de primer instar a concentraciones de 23 ng/mL de la toxina. Otros bioensayos indican que A. excentricus recombinante es tóxica para larvas de primer instar de las especies Aedes aegypti y Anopheles albimanus. Los ensayos de flotabilidad indican que A. excentricus no sedimenta hasta pasados 7 días, mientras que B. thuringiensis serovar. israelensis sedimenta al cabo de algunas horas. Palabras clave: Asticcacaulis excentricus, Bacillus thuringiensis, bacterias prostecadas, dengue, malaria.Bacillus thuringiensis cry genes encode for a diverse group of crystal-forming proteins that exhibit insecticidal activity towards dipteran, lepidopteran and coleopteran larvae. The effectiveness of

  14. Isolation and identification of some Bacillus thuringiensis strains with insecticidal activity against Ceratitis capitata

    International Nuclear Information System (INIS)

    Majdoub, Nihed

    2010-01-01

    The aims of the present work is to study the effect of toxins (delta-endotoxins), extracted from different strains of Bacillus thuringiensis on Ceratitis capitata, a devastating of citrus and fruit trees. Strains of B. thuringiensis were isolated from the mud of Sebket Sejoumi. Among 70 isolates tested, 15 showed a significant identicalness activity in which 5 isolates led to mortality rates ≥ 90 pour cent . These mortality rates are caused by endotoxins of B. thuringiensis. Analysis of proteins profiles of different isolates of B. thuringiensis revealed variability between them. The preliminary results of this study encourage us towards the characterization of the insecticidal activity produced by B. thuringiensis strains for large scale application.

  15. Relação entre toxicidade de proteínas Vip3Aa e sua capacidade de ligação a receptores intestinais de lepidópteros-praga

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    Suzana Cristina Marucci

    2015-08-01

    Full Text Available Resumo:O objetivo deste trabalho foi avaliar a toxicidade de novas proteínas Vip3Aa e sua capacidade de ligação a vesículas de membrana da microvilosidade apical (VMMA do intestino de lagartas neonatas de Spodoptera frugiperda, Anticarsia gemmatalise Heliothis virescens. Proteínas expressas pelos genes vip3Aa42 e vip3Aa43 mostraram-se tóxicas a S. frugiperda (CL50 de 78,2 e 113 ng cm-2, respectivamente e A. gemmatalis(CL50 de 239,2 e 57,5 ng cm-2, respectivamente, e pouco tóxicas a H. virescens (CL50>5.000 ng cm-2. Os ensaios de ligação às VMMA mostraram que as proteínas unem-se de forma efetiva aos receptores nas vesículas das espécies avaliadas, mas essa capacidade de ligação somente é efetiva na ativação da toxicidade para as populações avaliadas de S. frugiperdae A. gemmatalis.

  16. Occurrence of Natural Bacillus thuringiensis Contaminants and Residues of Bacillus thuringiensis-Based Insecticides on Fresh Fruits and Vegetables

    Science.gov (United States)

    Frederiksen, Kristine; Rosenquist, Hanne; Jørgensen, Kirsten; Wilcks, Andrea

    2006-01-01

    A total of 128 Bacillus cereus-like strains isolated from fresh fruits and vegetables for sale in retail shops in Denmark were characterized. Of these strains, 39% (50/128) were classified as Bacillus thuringiensis on the basis of their content of cry genes determined by PCR or crystal proteins visualized by microscopy. Random amplified polymorphic DNA analysis and plasmid profiling indicated that 23 of the 50 B. thuringiensis strains were of the same subtype as B. thuringiensis strains used as commercial bioinsecticides. Fourteen isolates were indistinguishable from B. thuringiensis subsp. kurstaki HD1 present in the products Dipel, Biobit, and Foray, and nine isolates grouped with B. thuringiensis subsp. aizawai present in Turex. The commercial strains were primarily isolated from samples of tomatoes, cucumbers, and peppers. A multiplex PCR method was developed to simultaneously detect all three genes in the enterotoxin hemolysin BL (HBL) and the nonhemolytic enterotoxin (NHE), respectively. This revealed that the frequency of these enterotoxin genes was higher among the strains indistinguishable from the commercial strains than among the other B. thuringiensis and B. cereus-like strains isolated from fruits and vegetables. The same was seen for a third enterotoxin, CytK. In conclusion, the present study strongly indicates that residues of B. thuringiensis-based insecticides can be found on fresh fruits and vegetables and that these are potentially enterotoxigenic. PMID:16672488

  17. Binding specificity of Bacillus thuringiensis Cry1Aa for purified, native Bombyx mori aminopeptidase N and cadherin-like receptors

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    Jenkins Jeremy L

    2001-10-01

    Full Text Available Abstract Background To better understand the molecular interactions of Bt toxins with non-target insects, we have examined the real-time binding specificity and affinity of Cry1 toxins to native silkworm (Bombyx mori midgut receptors. Previous studies on B. mori receptors utilized brush border membrane vesicles or purifed receptors in blot-type assays. Results The Bombyx mori (silkworm aminopeptidase N (APN and cadherin-like receptors for Bacillus thuringiensis insecticidal Cry1Aa toxin were purified and their real-time binding affinities for Cry toxins were examined by surface plasmon resonance. Cry1Ab and Cry1Ac toxins did not bind to the immobilized native receptors, correlating with their low toxicities. Cry1Aa displayed moderate affinity for B. mori APN (75 nM, and unusually tight binding to the cadherin-like receptor (2.6 nM, which results from slow dissociation rates. The binding of a hybrid toxin (Aa/Aa/Ac was identical to Cry1Aa. Conclusions These results indicate domain II of Cry1Aa is essential for binding to native B. mori receptors and for toxicity. Moreover, the high-affinity binding of Cry1Aa to native cadherin-like receptor emphasizes the importance of this receptor class for Bt toxin research.

  18. Binding specificity of Bacillus thuringiensis Cry1Aa for purified, native Bombyx mori aminopeptidase N and cadherin-like receptors

    Science.gov (United States)

    Jenkins, Jeremy L; Dean, Donald H

    2001-01-01

    Background To better understand the molecular interactions of Bt toxins with non-target insects, we have examined the real-time binding specificity and affinity of Cry1 toxins to native silkworm (Bombyx mori) midgut receptors. Previous studies on B. mori receptors utilized brush border membrane vesicles or purifed receptors in blot-type assays. Results The Bombyx mori (silkworm) aminopeptidase N (APN) and cadherin-like receptors for Bacillus thuringiensis insecticidal Cry1Aa toxin were purified and their real-time binding affinities for Cry toxins were examined by surface plasmon resonance. Cry1Ab and Cry1Ac toxins did not bind to the immobilized native receptors, correlating with their low toxicities. Cry1Aa displayed moderate affinity for B. mori APN (75 nM), and unusually tight binding to the cadherin-like receptor (2.6 nM), which results from slow dissociation rates. The binding of a hybrid toxin (Aa/Aa/Ac) was identical to Cry1Aa. Conclusions These results indicate domain II of Cry1Aa is essential for binding to native B. mori receptors and for toxicity. Moreover, the high-affinity binding of Cry1Aa to native cadherin-like receptor emphasizes the importance of this receptor class for Bt toxin research. PMID:11722800

  19. [Genes of insecticidal crystal proteins with dual specificity in Bacillus thuringiensis strains, isolated in the Crimea territory].

    Science.gov (United States)

    Rymar, S Iu; Isakova, I A; Kuznietsova, L M; Kordium, V A

    2006-01-01

    The insecticidal crystal proteins of 15 B. thuringiensis strains, isolated in the Crimea territory that are toxical for some Lepidoptera and Colorado potato beetle larvae were identified by PAGE electrophoresis. Ten strains produced the crystal proteins with high molecular weight (> 120 kD). PCR with use of broad specificity primers and DNA of these B. thuringiensis strains as template demonstrated the specific PCR products (1000 bp). Amplified DNA fragments were cloned and sequenced. The nucleotide sequence analysis revealed that the genomes of ten strains of B. thuringiensis carried Cry1B genes, which are responsible for production of the insecticidal crystal proteins with dual specificity. The influence of the solubilization conditions on the structure and toxicity of Cry1B protein for Colorado potato beetle larvae was shown. The dual toxicity of studied B. thuringiensis strains is explained by the Cry1B genes presence in their genomes. These strains may be used to develop the broad specificity bioinsecticides.

  20. Detecting authorized and unauthorized genetically modified organisms containing vip3A by real-time PCR and next-generation sequencing.

    Science.gov (United States)

    Liang, Chanjuan; van Dijk, Jeroen P; Scholtens, Ingrid M J; Staats, Martijn; Prins, Theo W; Voorhuijzen, Marleen M; da Silva, Andrea M; Arisi, Ana Carolina Maisonnave; den Dunnen, Johan T; Kok, Esther J

    2014-04-01

    The growing number of biotech crops with novel genetic elements increasingly complicates the detection of genetically modified organisms (GMOs) in food and feed samples using conventional screening methods. Unauthorized GMOs (UGMOs) in food and feed are currently identified through combining GMO element screening with sequencing the DNA flanking these elements. In this study, a specific and sensitive qPCR assay was developed for vip3A element detection based on the vip3Aa20 coding sequences of the recently marketed MIR162 maize and COT102 cotton. Furthermore, SiteFinding-PCR in combination with Sanger, Illumina or Pacific BioSciences (PacBio) sequencing was performed targeting the flanking DNA of the vip3Aa20 element in MIR162. De novo assembly and Basic Local Alignment Search Tool searches were used to mimic UGMO identification. PacBio data resulted in relatively long contigs in the upstream (1,326 nucleotides (nt); 95 % identity) and downstream (1,135 nt; 92 % identity) regions, whereas Illumina data resulted in two smaller contigs of 858 and 1,038 nt with higher sequence identity (>99 % identity). Both approaches outperformed Sanger sequencing, underlining the potential for next-generation sequencing in UGMO identification.

  1. Isolation of strains of Bacillus thuringiensis insecticidal biological activity against Ceratitis capitata

    International Nuclear Information System (INIS)

    Hmaied, Ezzedine; Ben Mbarek, Wael

    2010-01-01

    The present work is to study the effect of toxins (δ-endotoxins) extracted from strains of Bacillus thuringiensis isolated from the mud on the fly Sabkhat Dejoumi Ceratitis capitata, a pest of citrus and fruit trees. Among 51 isolated tested, 15 showed a very significant insecticidal activity, characterized by mortality rates exceeding 80 pour cent. These mortality rates are caused by endotoxins of Bt revealed variability between them. The preliminary results of this study encourage us towards the characterization of the insecticidal activity produced by strains of Bt for large scale application.

  2. Characterization of Baculovirus Insecticides Expressing Tailored Bacillus thuringiensis CryIA(b) Crystal Proteins

    NARCIS (Netherlands)

    Martens, John W M; Knoester, Marga; Weijts, Franci; Groffen, Sander J A; Hu, Zhihong; Bosch, Dirk; Vlak, Just M.

    1995-01-01

    Full-length, truncated, and mature forms of the CryIA(b) insecticidal crystal protein gene of Bacillus thuringiensis were engineered into the p10 locus of Autographa californica nuclear polyhedrosis virus (AcNPV). A signal sequence of Heliothis virescens juvenile hormone esterase was introduced at

  3. Transcriptome profiling of the intoxication response of Tenebrio molitor larvae to Bacillus thuringiensis Cry3Aa protoxin.

    Science.gov (United States)

    Oppert, Brenda; Dowd, Scot E; Bouffard, Pascal; Li, Lewyn; Conesa, Ana; Lorenzen, Marcé D; Toutges, Michelle; Marshall, Jeremy; Huestis, Diana L; Fabrick, Jeff; Oppert, Cris; Jurat-Fuentes, Juan Luis

    2012-01-01

    Bacillus thuringiensis (Bt) crystal (Cry) proteins are effective against a select number of insect pests, but improvements are needed to increase efficacy and decrease time to mortality for coleopteran pests. To gain insight into the Bt intoxication process in Coleoptera, we performed RNA-Seq on cDNA generated from the guts of Tenebrio molitor larvae that consumed either a control diet or a diet containing Cry3Aa protoxin. Approximately 134,090 and 124,287 sequence reads from the control and Cry3Aa-treated groups were assembled into 1,318 and 1,140 contigs, respectively. Enrichment analyses indicated that functions associated with mitochondrial respiration, signalling, maintenance of cell structure, membrane integrity, protein recycling/synthesis, and glycosyl hydrolases were significantly increased in Cry3Aa-treated larvae, whereas functions associated with many metabolic processes were reduced, especially glycolysis, tricarboxylic acid cycle, and fatty acid synthesis. Microarray analysis was used to evaluate temporal changes in gene expression after 6, 12 or 24 h of Cry3Aa exposure. Overall, microarray analysis indicated that transcripts related to allergens, chitin-binding proteins, glycosyl hydrolases, and tubulins were induced, and those related to immunity and metabolism were repressed in Cry3Aa-intoxicated larvae. The 24 h microarray data validated most of the RNA-Seq data. Of the three intoxication intervals, larvae demonstrated more differential expression of transcripts after 12 h exposure to Cry3Aa. Gene expression examined by three different methods in control vs. Cry3Aa-treated larvae at the 24 h time point indicated that transcripts encoding proteins with chitin-binding domain 3 were the most differentially expressed in Cry3Aa-intoxicated larvae. Overall, the data suggest that T. molitor larvae mount a complex response to Cry3Aa during the initial 24 h of intoxication. Data from this study represent the largest genetic sequence dataset for T. molitor

  4. Transcriptome profiling of the intoxication response of Tenebrio molitor larvae to Bacillus thuringiensis Cry3Aa protoxin.

    Directory of Open Access Journals (Sweden)

    Brenda Oppert

    Full Text Available Bacillus thuringiensis (Bt crystal (Cry proteins are effective against a select number of insect pests, but improvements are needed to increase efficacy and decrease time to mortality for coleopteran pests. To gain insight into the Bt intoxication process in Coleoptera, we performed RNA-Seq on cDNA generated from the guts of Tenebrio molitor larvae that consumed either a control diet or a diet containing Cry3Aa protoxin. Approximately 134,090 and 124,287 sequence reads from the control and Cry3Aa-treated groups were assembled into 1,318 and 1,140 contigs, respectively. Enrichment analyses indicated that functions associated with mitochondrial respiration, signalling, maintenance of cell structure, membrane integrity, protein recycling/synthesis, and glycosyl hydrolases were significantly increased in Cry3Aa-treated larvae, whereas functions associated with many metabolic processes were reduced, especially glycolysis, tricarboxylic acid cycle, and fatty acid synthesis. Microarray analysis was used to evaluate temporal changes in gene expression after 6, 12 or 24 h of Cry3Aa exposure. Overall, microarray analysis indicated that transcripts related to allergens, chitin-binding proteins, glycosyl hydrolases, and tubulins were induced, and those related to immunity and metabolism were repressed in Cry3Aa-intoxicated larvae. The 24 h microarray data validated most of the RNA-Seq data. Of the three intoxication intervals, larvae demonstrated more differential expression of transcripts after 12 h exposure to Cry3Aa. Gene expression examined by three different methods in control vs. Cry3Aa-treated larvae at the 24 h time point indicated that transcripts encoding proteins with chitin-binding domain 3 were the most differentially expressed in Cry3Aa-intoxicated larvae. Overall, the data suggest that T. molitor larvae mount a complex response to Cry3Aa during the initial 24 h of intoxication. Data from this study represent the largest genetic sequence

  5. Detection, cloning and bioinformatics analysis of vip1/vip2 genes ...

    African Journals Online (AJOL)

    Bio-insecticides based on the spore forming bacterium, Bacillus thuringiensis (Bt) have been used for commercial scale for the past 40 years. Bt is a Gram-positive soil bacterium that forms insecticidal crystal proteins (ICPs) during sporulation; it has been characterized as an insect pathogen. Vegetative insecticidal protein ...

  6. c-di-GMP Regulates Various Phenotypes and Insecticidal Activity of Gram-Positive Bacillus thuringiensis

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    Yang Fu

    2018-02-01

    Full Text Available C-di-GMP has been well investigated to play significant roles in the physiology of many Gram-negative bacteria. However, its effect on Gram-positive bacteria is less known. In order to more understand the c-di-GMP functions in Gram-positive bacteria, we have carried out a detailed study on the c-di-GMP-metabolizing enzymes and their physiological functions in Bacillus thuringiensis, a Gram-positive entomopathogenic bacterium that has been applied as an insecticide successfully. We performed a systematic study on the ten putative c-di-GMP-synthesizing enzyme diguanylate cyclases (DGCs and c-di-GMP-degrading enzyme phosphodiesterases (PDEs in B. thuringiensis BMB171, and artificially elevated the intracellular c-di-GMP level in BMB171 by deleting one or more pde genes. We found increasing level of intracellular c-di-GMP exhibits similar activities as those in Gram-negative bacteria, including altered activities in cell motility, biofilm formation, and cell-cell aggregation. Unexpectedly, we additionally found a novel function exhibited by the increasing level of c-di-GMP to promote the insecticidal activity of this bacterium against Helicoverpa armigera. Through whole-genome transcriptome profile analyses, we found that 4.3% of the B. thuringiensis genes were differentially transcribed when c-di-GMP level was increased, and 77.3% of such gene products are involved in some regulatory pathways not reported in other bacteria to date. In summary, our study represents the first comprehensive report on the c-di-GMP-metabolizing enzymes, their effects on phenotypes, and the transcriptome mediated by c-di-GMP in an important Gram-positive bacterium.

  7. Isolation of Bacillus thuringiensis from the state of Amazonas, in Brazil, and screening against Aedes aegypti (Diptera, Culicidae

    Directory of Open Access Journals (Sweden)

    Joelma Soares-da-Silva

    2015-03-01

    Full Text Available We investigated the use of Bacillus thuringiensis isolated in the state of Amazonas, in Brazil, for the biological control of the dengue vector Aedes aegypti. From 25 soil samples collected in nine municipalities, 484 bacterial colonies were obtained, 57 (11.78% of which were identified as B. thuringiensis. Six isolates, IBt-03, IBt-06, IBt-07, IBt-28, IBt-30, and BtAM-27 showed insecticidal activity, and only BtAM-27 presents the five genes investigated cry4Aa, cry4Ba, cry10Aa, cry11Aa, and cry11Ba. The IBt-07 and IBt- 28, with lower LC50 values, showed equal toxicity compared to the standards. The isolates of B. thuringiensis from Amazonas constitute potential new means of biological control for A. aegypti, because of their larvicidal activity and the possibility that they may also contain new combinations of toxins.

  8. Increased toxicity of Bacillus thuringiensis Cry3Aa against Crioceris quatuordecimpunctata, Phaedon brassicae and Colaphellus bowringi by a Tenebrio molitor cadherin fragment

    Science.gov (United States)

    BACKGROUND: Biopesticides containing Cry insecticidal proteins from the bacterium Bacillus thuringiensis (Bt) are effective against many lepidopteran pests, but there is a lack of Bt-based pesticides to efficiently control important coleopteran pests. Based on the reported increase of Bt toxin olig...

  9. Larvicidal activity of Bacillus thuringiensis var. israelensis Cry11Aa toxin against Haemonchus contortus.

    Science.gov (United States)

    DE Lara, Ana Paula DE Souza Stori; Lorenzon, Lucas Bigolin; Vianna, Ana Muñoz; Santos, Francisco Denis Souza; Pinto, Luciano Silva; Aires Berne, Maria Elisabeth; Leite, Fábio Pereira Leivas

    2016-10-01

    Effective control of gastrointestinal parasites is necessary in sheep production. The development of anthelmintics resistance is causing the available chemically based anthelmintics to become less effective. Biological control strategies present an alternative to this problem. In the current study, we tested the larvicidal effects of Bacillus thuringiensis var. israelensis Cry11Aa toxin against Haemonchus contortus larvae. Bacterial suspensions [2 × 108 colony-forming units (CFU) g-1 of the feces] of B. thuringiensis var. israelensis and recombinant Escherichia coli expressing Cry11Aa toxin were added to naturally H. contortus egg-contaminated feces. The larvae were quantified, and significant reductions of 62 and 81% (P var. israelensis and recombinant E. coli expressing Cry11Aa toxin were then orally administered to lambs naturally infected with H. contortus. Twelve hours after administration, feces were collected and submitted to coprocultures. Significant larvae reductions (P var. israelensis is a promising new class of biological anthelmintics for treating sheep against H. contortus.

  10. Identification of Bacillus thuringiensis Cry3Aa toxin domain II loop 1 as the binding site of Tenebrio molitor cadherin repeat CR12.

    Science.gov (United States)

    Zúñiga-Navarrete, Fernando; Gómez, Isabel; Peña, Guadalupe; Amaro, Itzel; Ortíz, Ernesto; Becerril, Baltazar; Ibarra, Jorge E; Bravo, Alejandra; Soberón, Mario

    2015-04-01

    Bacillus thuringiensis Cry toxins exert their toxic effect by specific recognition of larval midgut proteins leading to oligomerization of the toxin, membrane insertion and pore formation. The exposed domain II loop regions of Cry toxins have been shown to be involved in receptor binding. Insect cadherins have shown to be functionally involved in toxin binding facilitating toxin oligomerization. Here, we isolated a VHH (VHHA5) antibody by phage display that binds Cry3Aa loop 1 and competed with the binding of Cry3Aa to Tenebrio molitor brush border membranes. VHHA5 also competed with the binding of Cry3Aa to a cadherin fragment (CR12) that was previously shown to be involved in binding and toxicity of Cry3Aa, indicating that Cry3Aa binds CR12 through domain II loop 1. Moreover, we show that a loop 1 mutant, previously characterized to have increased toxicity to T. molitor, displayed a correlative enhanced binding affinity to T. molitor CR12 and to VHHA5. These results show that Cry3Aa domain II loop 1 is a binding site of CR12 T. molitor cadherin. Copyright © 2015 Elsevier Ltd. All rights reserved.

  11. High concentrations of protein test substances may have non-toxic effects on Daphnia magna: implications for regulatory study designs and ecological risk assessments for GM crops.

    Science.gov (United States)

    Raybould, Alan; Burns, Andrea; Hamer, Mick

    2014-01-01

    Laboratory testing for possible adverse effects of insecticidal proteins on non-target organisms (NTOs) is an important part of many ecological risk assessments for regulatory decision-making about the cultivation of insect-resistant genetically modified (IRGM) crops. To increase confidence in the risk assessments, regulatory guidelines for effects testing specify that representative surrogate species for NTOs are exposed to concentrations of insecticidal proteins that are in excess of worst-case predicted exposures in the field. High concentrations in effects tests are achieved by using protein test substances produced in microbes, such as Escherichia coli. In a study that exposed Daphnia magna to a single high concentration of a microbial test substance containing Vip3Aa20, the insecticidal protein in MIR162 maize, small reductions in growth were observed. These effects were surprising as many other studies strongly suggest that the activity of Vip3Aa20 is limited to Lepidoptera. A plausible explanation for the effect on growth is that high concentrations of test substance have a non-toxic effect on Daphnia, perhaps by reducing its feeding rate. A follow-up study tested that hypothesis by exposing D. magna to several concentrations of Vip3Aa20, and a high concentration of a non-toxic protein, bovine serum albumin (BSA). Vip3Aa20 and BSA had sporadic effects on the reproduction and growth of D. magna. The pattern of the effects suggests that they result from non-toxic effects of high concentrations of protein, and not from toxicity. The implications of these results for regulatory NTO effects testing and ERA of IRGM crops are discussed.

  12. Bacillus thuringiensis Cry3Aa protoxin intoxication of Tenebrio molitor induces widespread changes in the expression of serine peptidase transcripts.

    Science.gov (United States)

    Oppert, Brenda; Martynov, Alexander G; Elpidina, Elena N

    2012-09-01

    The yellow mealworm, Tenebrio molitor, is a pest of stored grain products and is sensitive to the Bacillus thuringiensis (Bt) Cry3Aa toxin. As digestive peptidases are a determining factor in Cry toxicity and resistance, we evaluated the expression of peptidase transcripts in the midgut of T. molitor larvae fed either a control or Cry3Aa protoxin diet for 24 h (RNA-Seq), or in larvae exposed to the protoxin for 6, 12, or 24 h (microarrays). Cysteine peptidase transcripts (9) were similar to cathepsins B, L, and K, and their expression did not vary more than 2.5-fold in control and Cry3Aa-treated larvae. Serine peptidase transcripts (48) included trypsin, chymotrypsin and chymotrypsin-like, elastase 1-like, and unclassified serine peptidases, as well as homologs lacking functional amino acids. Highly expressed trypsin and chymotrypsin transcripts were severely repressed, and most serine peptidase transcripts were expressed 2- to 15-fold lower in Cry3Aa-treated larvae. Many serine peptidase and homolog transcripts were found only in control larvae. However, expression of a few serine peptidase transcripts was increased or found only in Cry3Aa-treated larvae. Therefore, Bt intoxication significantly impacted the expression of serine peptidases, potentially important in protoxin processing, while the insect maintained the production of critical digestive cysteine peptidases. Published by Elsevier Inc.

  13. A Tenebrio molitor GPI-anchored alkaline phosphatase is involved in binding of Bacillus thuringiensis Cry3Aa to brush border membrane vesicles.

    Science.gov (United States)

    Zúñiga-Navarrete, Fernando; Gómez, Isabel; Peña, Guadalupe; Bravo, Alejandra; Soberón, Mario

    2013-03-01

    Bacillus thuringiensis Cry toxins recognizes their target cells in part by the binding to glycosyl-phosphatidyl-inositol (GPI) anchored proteins such as aminopeptidase-N (APN) or alkaline phosphatases (ALP). Treatment of Tenebrio molitor brush border membrane vesicles (BBMV) with phospholipase C that cleaves out GPI-anchored proteins from the membranes, showed that GPI-anchored proteins are involved in binding of Cry3Aa toxin to BBMV. A 68 kDa GPI-anchored ALP was shown to bind Cry3Aa by toxin overlay assays. The 68 kDa GPI-anchored ALP was preferentially expressed in early instar larvae in comparison to late instar larvae. Our work shows for the first time that GPI-anchored ALP is important for Cry3Aa binding to T. molitor BBMV suggesting that the mode of action of Cry toxins is conserved in different insect orders. Copyright © 2012 Elsevier Inc. All rights reserved.

  14. Evolution of Bacillus thuringiensis Cry toxins insecticidal activity.

    Science.gov (United States)

    Bravo, Alejandra; Gómez, Isabel; Porta, Helena; García-Gómez, Blanca Ines; Rodriguez-Almazan, Claudia; Pardo, Liliana; Soberón, Mario

    2013-01-01

    Insecticidal Cry proteins produced by Bacillus thuringiensis are use worldwide in transgenic crops for efficient pest control. Among the family of Cry toxins, the three domain Cry family is the better characterized regarding their natural evolution leading to a large number of Cry proteins with similar structure, mode of action but different insect specificity. Also, this group is the better characterized regarding the study of their mode of action and the molecular basis of insect specificity. In this review we discuss how Cry toxins have evolved insect specificity in nature and analyse several cases of improvement of Cry toxin action by genetic engineering, some of these examples are currently used in transgenic crops. We believe that the success in the improvement of insecticidal activity by genetic evolution of Cry toxins will depend on the knowledge of the rate-limiting steps of Cry toxicity in different insect pests, the mapping of the specificity binding regions in the Cry toxins, as well as the improvement of mutagenesis strategies and selection procedures. © 2012 The Authors. Microbial Biotechnology © 2012 Society for Applied Microbiology and Blackwell Publishing Ltd.

  15. 78 FR 69849 - Issuance of an Experimental Use Permit

    Science.gov (United States)

    2013-11-21

    ... http://www.regulations.gov or at the Office of Pesticide Programs Regulatory Public Docket (OPP Docket... Dv49 double stranded RNA (dsRNA) suppression cassette in combination with Bacillus thuringiensis (Bt....105, Cry2Ab2, Vip3Aa20, Dv49 dsRNA, Cry3Bb1, Cry34Ab1, Cry35Ab1, eCry3.1Ab, respectively) are to be...

  16. Evaluation of the time-concentration-mortality responses of Plutella xylostella larvae to the interaction of Isaria fumosorosea with the insecticides beta-cypermethrin and Bacillus thuringiensis.

    Science.gov (United States)

    Nian, Xiao-ge; He, Yu-rong; Lu, Li-hua; Zhao, Rui

    2015-02-01

    Entomopathogenic fungi are potential candidates for controlling Plutella xylostella, a cosmopolitan pest of crucifers. In this study, bioassays were conducted to evaluate the interaction between Isaria fumosorosea and sublethal doses of two insecticides, beta-cypermethrin and Bacillus thuringiensis, against P. xylostella. Data of each assay were in good agreement with the time-concentration-mortality model, indicating a strong dependence of the fungus and insecticide interaction on both concentration and post-exposure time. Using beta-cypermethrin 58-116 µg mL(-1) or B. thuringiensis 222.5-890 µg mL(-1) with the fungus significantly enhanced fungal efficacy. The LC50 values of the fungus declined over a 1-7 day period after exposure, and the LT50 values decreased with increasing concentration. Based on LC50 or LC90 estimates, synergism between the fungus and beta-cypermethrin resulted in a 2.7-28.3-fold reduction in LC50 values and a 12.1-19.6-fold reduction in LC90 values, while synergism of the fungus with B. thuringiensis led to a 2.4-385.0-fold reduction in LC50 values and a 4.4-151.7-fold reduction in LC90 values. Results show that sublethal doses of B. thuringiensis and beta-cypermethrin can synergise I. fumosorosea activity on P. xylostella, suggesting that combination of I. fumosorosea with the two insecticides might offer an integrated approach to controlling P. xylostella in practice. © 2014 Society of Chemical Industry. © 2014 Society of Chemical Industry.

  17. Mobility of adsorbed Cry1Aa insecticidal toxin from Bacillus thuringiensis (Bt) on montmorillonite measured by fluorescence recovery after photobleaching (FRAP)

    Science.gov (United States)

    Helassa, Nordine; Daudin, Gabrielle; Noinville, Sylvie; Janot, Jean-Marc; Déjardin, Philippe; Staunton, Siobhán; Quiquampoix, Hervé

    2010-06-01

    The insecticidal toxins produced by genetically modified Bt crops are introduced into soil through root exudates and tissue decomposition and adsorb readily on soil components, especially on clays. This immobilisation and the consequent concentration of the toxins in "hot spots" could increase the exposure of soil organisms. Whereas the effects on non-target organisms are well documented, few studies consider the migration of the toxin in soil. In this study, the residual mobility of Bt Cry1Aa insecticidal toxin adsorbed on montmorillonite was assessed using fluorescence recovery after photobleaching (FRAP). This technique, which is usually used to study dynamics of cytoplasmic and membrane molecules in live cells, was applied for the first time to a protein adsorbed on a finely divided swelling clay mineral, montmorillonite. No mobility of adsorbed toxin was observed at any pH and at different degrees of surface saturation.

  18. Evaluation of Cytotoxicity, Genotoxicity and Hematotoxicity of the Recombinant Spore-Crystal Complexes Cry1Ia, Cry10Aa and Cry1Ba6 from Bacillus thuringiensis in Swiss Mice

    Directory of Open Access Journals (Sweden)

    Ingrid de Souza Freire

    2014-09-01

    Full Text Available The insecticidal properties of Cry-endotoxins from Bacillus thuringiensis (Bt have long been used as spore-crystals in commercial spray formulations for insect control. Recently, some Bt-endotoxin genes have been cloned in many different plants. Toxicological evaluations of three spore-crystal endotoxins, BtCry1Ia, BtCry10Aa and BtCry1Ba6 from B. thuringiensis, were carried out on mice to understand their adverse effects on hematological systems and on genetic material. These three spore-crystals have shown toxic activity to the boll weevil, which is one of the most aggressive pests of the cotton crop. Cry1Ia, Cry10Aa and Cry1Ba6 did not increase the micronucleus frequency in the peripheral erythrocytes of mice and did not cause changes in the frequency of polychromatic erythrocytes. However, some hematologic disburbances were observed, specifically related to Cry1Ia and Cry1Ba6, respectively, for the erythroid and lymphoid lineage. Thus, although the profile of such adverse side effects can be related to their high level of exposure, which is not commonly found in the environment, results showed that these Bt spore-crystals were not harmless to mice, indicating that each spore-crystal endotoxin presents a characteristic profile of toxicity and might be investigated individually.

  19. Improved insecticidal toxicity by fusing Cry1Ac of Bacillus thuringiensis with Av3 of Anemonia viridis.

    Science.gov (United States)

    Yan, Fu; Cheng, Xing; Ding, Xuezhi; Yao, Ting; Chen, Hanna; Li, Wenping; Hu, Shengbiao; Yu, Ziquan; Sun, Yunjun; Zhang, Youming; Xia, Liqiu

    2014-05-01

    Av3, a neurotoxin of Anemonia viridis, is toxic to crustaceans and cockroaches but inactive in mammals. In the present study, Av3 was expressed in Escherichia coli Origami B (DE3) and purified by reversed-phase liquid chromatography. The purified Av3 was injected into the hemocoel of Helicoverpa armigera, rendering the worm paralyzed. Then, Av3 was expressed alone or fusion expressed with the Cry1Ac in acrystalliferous strain Cry(-)B of Bacillus thuringiensis. The shape of Cry1Ac was changed by fusion with Av3. The expressed fusion protein, Cry1AcAv3, formed irregular rhombus- or crescent-shaped crystalline inclusions, which is quite different from the shape of original Cry1Ac crystals. The toxicity of Cry1Ac was improved by fused expression. Compared with original Cry1Ac expressed in Cry(-)B, the oral toxicity of Cry1AcAv3 to H. armigera was elevated about 2.6-fold. No toxicity was detected when Av3 was expressed in Cry(-)B alone. The present study confirmed that marine toxins could be used in bio-control and implied that fused expression with other insecticidal proteins could be an efficient way for their application.

  20. [Transformation and expression of specific insecticide gene Bt cry3A in resident endogenetic bacteria isolated from Apriona germari (Hope) larvae intestines].

    Science.gov (United States)

    Zhongkang, Wang; Wei, He; Guoxiong, Peng; Yuxian, Xia; Qiang, Li; Youping, Yin

    2008-09-01

    Transforming the specific insecticidal gene Bt cry3A into the dominant resident endogenetic bacteria in intestines of Apriona germari (Hope) larvae to construct transgenic bacteria that can colonize and express the insecticidal gene Bt cry3A perfectly in intestines of Apriona germari (Hope) larvae. We isolated and identified the dominant resident endogenetic bacteria by traditional methods and molecular method based of 16S rDNA analysis. Two Escherichia coli--Bacillus thuringiensis shuttle plasmid pHT305a and pHT7911 which contained specific insecticidal gene Bt cry3A were transformed into two resident endogenetic bacteria Brevibacillus brevis Ag12 and Bacillus thuringiensis Ag13 isolated from A. germari larvae intestines respectively by electro-transformation. Eighteen species of bacteria have isolated and identified from Apriona germari larvae intestines and two of them (Brevibacillus brevis Ag12 and Bacillus thuringiensis Ag13) were selected as starting bacteria to recieve the Bt cry3A. The 4 transgenic engineering strains Ag12-7911, Ag12-305a, Ag13-7911 and Ag13-305a were obtained successfully and validated by testing the plasmid stability in recombinants, transformants vegetal properties, crystal poisonous protein observation, expressional protein SDS-PAGE. The Bt cry3A gene had been transformed into Brevibacillus brevis and Bacillus thuringiensis. Both bioassay and examination of the engineering strains in intestines after feeding them to larvae showed that all these transformant strains (Brevibacillus brevis Ag12-305a, Bacillus thurigiensis Ag13-305a, Brevibacillus brevis Ag12-7911 and Bacillus thurigiensis Ag13-7911) could colonize and express 65 kDa protoxin in intestines of A. germari larvae and had insecticidal activity. We obtained four transgenic bacteria that can colonize and express the target insecticide gene Bt cry3A in A. germari larvae. They may be developed as a new insecticide.

  1. Construction and characterization of the interdomain chimeras using Cry11Aa and Cry11Ba from Bacillus thuringiensis and identification of a possible novel toxic chimera.

    Science.gov (United States)

    Sun, Yunjun; Zhao, Qiang; Zheng, Dasheng; Ding, Xuezhi; Wang, Jingfang; Hu, Quanfang; Yuan, Zhiming; Park, Hyun-Woo; Xia, Liqiu

    2014-01-01

    Three structural domains of mosquitocidal Cry11Aa and Cry11Ba from Bacillus thuringiensis were exchanged to produce interdomain chimeras [BAA (11Ba/11Aa/11Aa), ABA (11Aa/11Ba/11Aa), AAB (11Aa/11Aa/11Ba), ABB (11Aa/11Ba/11Ba), BAB (11Ba/11Aa/11Ba), BBA (11Ba/11Ba/11Aa]. Chimeras BAB, BAA, BBA, and AAB formed inclusion bodies in the crystal-negative B. thuringiensis host and produced expected protein bands on SDS-PAGE gel. However, no inclusion body or target protein could be found for chimeras ABA and ABB. In bioassays using the fourth-instar larvae of Culex quinquefasciatus and Aedes aegypti, AAB had ~50 % lethal concentrations of 4.8 and 2.2 μg ml(-1), respectively; however, the rest of chimeras were not toxic. This study thus helps to understand the domain-function relationships of the Cry11Aa and Cry11Ba toxins. The toxic chimera, AAB, might be a candidate for mosquito control as its amino acid sequence is different from the two parental toxins.

  2. Presence and significance of Bacillus thuringiensis Cry proteins associated with the Andean weevil Premnotrypes vorax (Coleoptera: Curculionidae

    Directory of Open Access Journals (Sweden)

    SilvioAlejandro López-Pazos

    2009-12-01

    Full Text Available The Andean weevil Premnotrypes vorax represents an important cause of damage to Colombian potato crops. Due to the impact of this plague on the economy of the country, we searched for new alternatives for its biological control, based on the entomopathogenic bacteria Bacillus thuringiensis. A total of 300 B. thuringiensis strains obtained from potato plantations infested with P. vorax were analyzed through crystal morphology, SDS-PAGE, PCR and bioassays. We used site- directed mutagenesis to modify the Cry3Aa protein. Most of the B. thuringiensis isolates had a bipyramidal crystal morphology. SDS-PAGE analyses had seven strains groups with σ-endotoxins from 35 to 135 kDa. The genes cry 2 and cry 1 were significantly more frequent in the P. vorax habitat (PCR analyses. Three mutant toxins, 1 (D354E, 2 (R345A, ∆Y350, ∆Y351, and 3 (Q482A, S484A, R485A, were analyzed to assess their activity against P. vorax larvae. Toxicity was low, or absent, against P. vorax for isolates, wild type cry 3Aa and cry 3Aa mutants. The genetic characterization of the collection provides opportunities for the selection of strains to be tested in bioassays against other insect pests of agricultural importance, and for designing Cry proteins with improved insecticidal toxicity. Rev. Biol. Trop. 57 (4: 1235-1243. Epub 2009 December 01.El gorgojo andino Premnotrypes vorax es una causa importante de daño en los cultivos colombianos de este tubérculo. Debido al impacto que esta plaga tiene sobre la economía del país, nos interesamos en buscar alternativas nuevas para el control biológico de P. vorax, basadas en la bacteria entomopatógena Bacillus thuringiensis. Se recolectaron un total de 300 cepas de B. thuringiensis a partir de plantaciones de papa infestadas con P. vorax, las cuales fueron analizadas por medio de la morfología del cristal, SDS-PAGE, PCR y ensayos biológicos. La mayoría de los aislamientos de B. thuringiensis presentaron cristales

  3. Isolation and characterization of Bacillus thuringiensis from soils in ...

    African Journals Online (AJOL)

    Bioassays were used to test the insecticidal activity of B. thuringiensis strains ... of crystal protein genes, 7 tested positive for cry 4, cry 11, and cyt toxin genes. ... mosquitocidal cry and cyt genes in Bacillus thuringiensis subsp. israelensis.

  4. Prevalence and Toxin Characteristics of Bacillus thuringiensis Isolated from Organic Vegetables.

    Science.gov (United States)

    Kim, Jung-Beom; Choi, Ok-Kyung; Kwon, Sun-Mok; Cho, Seung-Hak; Park, Byung-Jae; Jin, Na Young; Yu, Yong Man; Oh, Deog-Hwan

    2017-08-28

    The prevalence and toxin characteristics of Bacillus thuringiensis isolated from 39 organic vegetables were investigated. B. thuringiensis was detected in 30 out of the 39 organic vegetables (76.9%) with a mean value of 2.60 log CFU/g. Twenty-five out of the 30 B. thuringiensis isolates (83.3%) showed insecticidal toxicity against Spodoptera exigua . The hblCDA, nheABC , and entFM genes were found to be the major toxin genes, but the ces gene was not detected in any of the tested B. thuringiensis isolates. The hemolysin BL enterotoxin was detected in all 30 B. thuringiensis isolates (100%). The non-hemolytic enterotoxin complex was found in 27 out of 30 B. thuringiensis isolates (90.0%). The B. thuringiensis tested in this study had similar toxin gene characteristics to B. cereus , which possessed more than one toxin gene. B. thuringiensis could have the potential risk of foodborne illness based on the toxin genes and toxin-producing ability.

  5. Persistence of Bt Bacillus thuringiensis Cry1Aa toxin in various soils determined by physicochemical reactions

    Science.gov (United States)

    Helassa, N.; Noinville, S.; Déjardin, P.; Janot, J. M.; Quiquampoix, H.; Staunton, S.

    2009-04-01

    Insecticidal Cry proteins from the soil bacterium, Bacillus thuringiensis (Bt) are produced by a class of genetically modified (GM) crops, and released into soils through root exudates and upon decomposition of residues. In contrast to the protoxin produced by the Bacillus, the protein produced in GM crops does not require activation in insect midguts and thereby potentially looses some of its species specificity. Although gene transfer and resistance emergence phenomena are well documented, the fate of these toxins in soil has not yet been clearly elucidated. Cry proteins, in common with other proteins, are adsorbed on soils and soil components. Adsorption on soil, and the reversibility of this adsorption is an important aspect of the environmental behaviour of these toxins. The orientation of the molecule and conformational changes on surfaces may modify the toxicity and confer some protection against microbial degradation. Adsorption will have important consequences for both the risk of exposition of non target species and the acquisition of resistance by target species. We have adopted different approaches to investigate the fate of Cry1Aa in soils and model minerals. In each series of experiments we endeavoured to maintain the protein in a monomeric form (pH above 6.5 and a high ionic strength imposed with 150 mM NaCl). The adsorption and the desorbability of the Cry1Aa Bt insecticidal protein were measured on two different homoionic clays: montmorillonite and kaolinite. Adsorption isotherms obtained followed a low affinity interaction for both clays and could be fitted using the Langmuir equation. Binding of the toxin decreased as the pH increased from 6.5 (close to the isoelectric point) to 9. Maximum adsorption was about 40 times greater on montmorillonite (1.71 g g-1) than on kaolinite (0.04 g g-1) in line with the contrasting respective specific surface areas of the minerals. Finally, some of the adsorbed toxin was desorbed by water and more, about 36

  6. Monoclonal Antibody Analysis and Insecticidal Spectrum of Three Types of Lepidopteran-Specific Insecticidal Crystal Proteins of Bacillus thuringiensis

    Science.gov (United States)

    Höfte, Herman; Van Rie, Jeroen; Jansens, Stefan; Van Houtven, Annemie; Vanderbruggen, Hilde; Vaeck, Mark

    1988-01-01

    We have investigated the protein composition and the insecticidal spectrum of crystals of 29 Bacillus thuringiensis strains active against lepidopteran larvae. All crystals contained proteins of 130 to 140 kilodaltons (kDa) which could be grouped into three types by the molecular weight of the protoxin and the trypsin-activated core fragment. Proteins of the three types showed a characteristic insecticidal spectrum when tested against five lepidopteran species. Type A crystal proteins were protoxins of 130 or 133 kDa, which were processed into 60-kDa toxins by trypsin. Several genes encoding crystal proteins of this type have been cloned and sequenced earlier. They are highly conserved in the N-terminal half of the toxic fragment and were previously classified in three subtypes (the 4.5-, 5.3-, and 6.6-kilobase subtypes) based on the restriction map of their genes. The present study shows that different proteins of these three subtypes were equally toxic against Manduca sexta and Pieris brassicae and had no detectable activity against Spodoptera littoralis. However, the 4.5-, 5.3-, and 6.6-kilobase subtypes differed in their toxicity against Heliothis virescens and Mamestra brassicae. Type B crystal proteins consisted of 140-kDa protoxins with a 55-kDa tryptic core fragment. These were only active against one of the five insect species tested (P. brassicae). The protoxin and the trypsin-activated toxin of type C were 135- and 63-kDa proteins, respectively. Proteins of this type were associated with high toxicity against S. littoralis and M. brassicae. A panel of 35 monoclonal antibodies was used to compare the structural characteristics of crystal proteins of the three different types and subtypes. Each type of protein could be associated with a typical epitope structure, indicating an unambiguous correlation between antigenic structure and insect specificity. Images PMID:16347711

  7. Structural studies of {delta}-endotoxin Cry 1 C from Bacillus thuringiensis

    Energy Technology Data Exchange (ETDEWEB)

    Guimaraes, B.G.; Garratt, R.C.; Oliva, G. [Sao Paulo Univ., Sao Carlos, SP (Brazil). Inst. de Fisica; Lemos, M.V.F. [UNESP, Jaboticabal, SP (Brazil). Dept. de Biologia Aplicada Agropecuaria; Arantes, O.M.N. [Universidade Estadual de Londrina, PR (Brazil). Dept. de Biologia Geral

    1996-12-31

    Full text. The {delta}-endotoxins are a family of crystal protein by a soil bacterium, Bacillus thuringiensis. The study of these proteins has been of great interest due to their highly specific activity against insects of the orders Lepidoptera, Diptera and Coleoptera. Thus, the {delta}a-endotoxins have been used for more than two decades as biological insecticides to control agricultural pests and, more recently, insects vectors of some diseases. The knowledge of their three-dimensional structures is very important to understand their mechanism of action and their high specificity. To date, the structure of only three proteins of the {delta}-endotoxins family have been reported: Cry3A, a coleopteran-specific toxin (beetle toxin){sup 1}, Cry1Aa, a lepidopteran-specific toxin (butterfly toxin){sup 2} and CytB, a dipteran-specific toxin (mosquito toxin){sup 3} Our work is aimed at the determination of the crystallographic structure by X-ray diffraction of {delta}-endotoxin Cry1C, also toxic to insects of the Lepidoptera order but towards families other than those affected by Cry1Aa. A comparison between these structures may lead to important conclusions about the reasons for the specificity and would allow the planning of mutants with more efficient activity. The cry1C gene was cloned into an adequate vector and expressed in an acrystalliferous B. thuringiensis strain. After cell culture and sporulation the microcrystals of Cry1C were separated by ultra-centrifugation in sacharose. The protoxin inclusion bodies were activated by commercial trpsin and the protease-resistant core was purified by anion-exchange chromatography. Crystallization experiments are being conducted in order to obtain single crystals suitable for diffraction measurements. We intend to use the Protein Crystallograph Station of the LNLS to collect data as soon as it is available and we have suitable crystals. (author) 3 refs.

  8. Preferential protection of domains ii and iii of bacillus thuringiensis cry1aa toxin by brush border membrane vesicles

    OpenAIRE

    Hussain, Syed-Rehan A.; Flórez, Álvaro M.; Dean, Donald H.; Alzate, Óscar

    2011-01-01

    Título español: Protección preferencial de los dominios II y III de la toxina Cry1Aa de Bacillus thuringiensis en Vesículas de Membrana de Borde de Cepillo Abstract The surface exposed Leucine 371 on loop 2 of domain II, in Cry1Aa toxin, was mutated to Lysine to generate the trypsin-sensitive mutant, L371K. Upon trypsin digestion L371K is cleaved into approximately 37 and 26 kDa fragments. These are separable on SDS-PAGE, but remain as a single molecule of 65 kDa upon purification by ...

  9. Preferential Protection of Domains II and III of Bacillus thuringiensis Cry1Aa Toxin by Brush Border Membrane Vesicles

    OpenAIRE

    Syed-Rehan A. Hussain; Álvaro M. Flórez; Donald H. Dean; Óscar Alzate

    2011-01-01

    Título español: Protección preferencial de los dominios II y III de la toxina Cry1Aa de Bacillus thuringiensis en Vesículas de Membrana de Borde de Cepillo Abstract The surface exposed Leucine 371 on loop 2 of domain II, in Cry1Aa toxin, was mutated to Lysine to generate the trypsin-sensitive mutant, L371K. Upon trypsin digestion L371K is cleaved into approximately 37 and 26 kDa fragments. These are separable on SDS-PAGE, but remain as a single molecule of 65 kDa upon purification by ...

  10. Characterization of lepidopteran-specific cry1 and cry2 gene harbouring native Bacillus thuringiensis isolates toxic against Helicoverpa armigera

    Directory of Open Access Journals (Sweden)

    Showkat Ahmad Lone

    2017-09-01

    Full Text Available Bacillus thuringiensis (Bt based biopesticides are feasible alternatives to chemical pesticides. Here, we present the distribution of lepidopteran-specific cry1 and cry2 genes in native B. thuringiensis. Forty four out of 86 colonies were found to harbour crystals by phase contrast microscopy exhibiting a Bt index of 0.51. PCR analysis resulted in the amplification of cry1 in 24 and cry2 in 14 isolates. Twelve of the isolates showed presence of both cry1 and cry2, while 18 isolates did not show presence of either of the genes. Toxicity screening using spore-crystal mixtures against 2nd instar larvae of Helicoverpa armigera revealed that the isolates (50% were either mildly toxic or not toxic (36.36%, and only 13.63% were toxic. The results are interesting, particularly so because the same isolates were previously reported to contain lepidopteran specific vip3A genes also, hence can complement the toxicity of the isolates harbouring vip3A genes.

  11. Bacillus thuringiensis and its application in agriculture | Ali | African ...

    African Journals Online (AJOL)

    Presently, a number of approaches to pest control via genetic engineering have been developed and genetically engineered crops expressing insecticidal characteristics are under cultivation for the last 15 years. Use of Bacillus thuringiensis genes encoding o̅ endotoxins with insecticidal characteristics is the major ...

  12. Diversity of Bacillus thuringiensis strains isolated from coffee plantations infested with the coffee berry borer Hypothenemus hampei

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    Glen Arrieta

    2004-09-01

    Full Text Available The coffee berry borer Hypothenemus hampei Ferrari (Coleoptera: Scolytidae was first reported infecting Costa Rican coffee plantations in the year 2000. Due to the impact that this plague has in the economy of the country, we were interested in seeking new alternatives for the biological control of H. hampei, based on the entomopathogenic bacteria Bacillus thuringiensis. Atotal of 202 B. thuringiensis isolates obtained from Costa Rican coffee plantations infested with H. hampei were analyzed through crystal morphology of the crystal inclusions and SDS-PAGE of d-endotoxins, while 105 strains were further evaluated by PCR for the presence cry, cyt and vip genes. Most of the Bt strains showed diverse crystal morphologies: pleomorphic (35%, oval (37%, bipyramidal (3%, bipyramidal and oval (12%, bipyramidal, oval and pleomorphic (10% and bipyramidal, oval and cubic (3%. The SDS-PAGE analyses of the crystal preparations showed five strains with delta -endotoxin from 20 to 40 kDa, six from 40 to 50 kDa, seven from 50 to 60 kDa, 19 from 60 to 70 kDa, 29 from 70 to 100 kDa and 39 from 100-145 kDa. PCR analyses demonstrated that the collection showed diverse cry genes profiles having several genes per strain: 78 strains contained the vip3 gene, 82 the cry2 gene, 45 the cry1 and 29 strains harbored cry3-cry7 genes. A total of 13 strains did not amplified with any of the cry primers used: cry1, cry2, cry37, cry5, cry11, cry12 and cry14. Forty-three different genetic profiles were found, mainly due to the combination of cry1A genes with other cry and vip genes. The genetic characterization of the collection provides opportunities for the selection of strains to be tested in bioassays against H. hampei and other insect pests of agricultural importance. Rev. Biol. Trop. 52(3: 757-764. Epub 2004 Dic 15.En el año 2000 se reportó por primera vez la principal plaga del cafeto, conocida como broca (Hypothenemus hampei Ferrari (Coleoptera: Scolitidae en

  13. Ultra-violet-resistant mutants of Bacillus thuringiensis

    Energy Technology Data Exchange (ETDEWEB)

    Jones, D R; Karunakaran, V [Polytechnic of Central London (UK). Faculty of Engineering and Science, School of Biological and Health Sciences; Burges, H D [Institute of Horticultural Research, Littlehampton (UK); Hacking, A J [Reading Univ. (UK). Dextra Labs.Ltd.

    1991-06-01

    One of the main disadvantages of using Bacillus thuringiensis as an insecticide is that the spore and crystal preparations applied to foliage are readily washed away by rain and are inactivated by sunlight. Spores from some strains of B. thuringiensis have been shown to be highly sensitive to u.v. light. This study has demonstrated how mutants with increased resistance to u.v., isolated by successive rounds of u.v. irradiation, and additionally with increased specific pathogenicity can be isolated. These techniques should be applied to strains that are frequently used in the industrial production of B.thuringiensis toxin. (author).

  14. Ultra-violet-resistant mutants of Bacillus thuringiensis

    International Nuclear Information System (INIS)

    Jones, D.R.; Karunakaran, V.; Hacking, A.J.

    1991-01-01

    One of the main disadvantages of using Bacillus thuringiensis as an insecticide is that the spore and crystal preparations applied to foliage are readily washed away by rain and are inactivated by sunlight. Spores from some strains of B. thuringiensis have been shown to be highly sensitive to u.v. light. This study has demonstrated how mutants with increased resistance to u.v., isolated by successive rounds of u.v. irradiation, and additionally with increased specific pathogenicity can be isolated. These techniques should be applied to strains that are frequently used in the industrial production of B.thuringiensis toxin. (author)

  15. Characterization of parasporin gene harboring Indian isolates of Bacillus thuringiensis

    OpenAIRE

    Lenina, N. K.; Naveenkumar, A.; Sozhavendan, A. E.; Balakrishnan, N.; Balasubramani, V.; Udayasuriyan, V.

    2013-01-01

    Bacillus thuringiensis (Bt) is popularly known as insecticidal bacterium. However, non-insecticidal Bt strains are more extensively available in natural environment than the insecticidal ones. Parasporin (PS) is a collection of genealogically heterogeneous Cry proteins synthesized in non-insecticidal isolates of Bt. An important character generally related with PS proteins is their strong cytocidal activity preferentially on human cancer cells of various origins. Identification and characteri...

  16. A theoretical model of the tridimensional structure of Bacillus thuringiensis subsp. medellin Cry 11Bb toxin deduced by homology modelling

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    Gutierrez Pablo

    2001-01-01

    Full Text Available Cry11Bb is an insecticidal crystal protein produced by Bacillus thuringiensis subsp. medellin during its stationary phase; this ¶-endotoxin is active against dipteran insects and has great potential for mosquito borne disease control. Here, we report the first theoretical model of the tridimensional structure of a Cry11 toxin. The tridimensional structure of the Cry11Bb toxin was obtained by homology modelling on the structures of the Cry1Aa and Cry3Aa toxins. In this work we give a brief description of our model and hypothesize the residues of the Cry11Bb toxin that could be important in receptor recognition and pore formation. This model will serve as a starting point for the design of mutagenesis experiments aimed to the improvement of toxicity, and to provide a new tool for the elucidation of the mechanism of action of these mosquitocidal proteins.

  17. Characterization of a Chitin-Binding Protein from Bacillus thuringiensis HD-1.

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    Naresh Arora

    Full Text Available Strains of Bacillus thuringiensis produce insecticidal proteins. These strains have been isolated from diverse ecological niches, such as soil, phylloplane, insect cadavers and grain dust. To effectively propagate, these strains produce a range of molecules that facilitate its multiplication in a competing environment. In this report, we have examined synthesis of a chitin-binding protein and evaluated its effect on fungi encountered in environment and its interaction with insecticidal proteins synthesized by B. thuringiensis. The gene encoding chitin-binding protein has been cloned and expressed. The purified protein has been demonstrated to interact with Cry insecticidal protein, Cry1Ac by Circular Dichrosim spectroscopy (CD and in vitro pull down assays. The chitin-binding protein potentiates insecticidal activity of bacillar insecticidal protein, Cry1Ac. Further, chitin-binding protein was fungistatic against several soil fungi. The chitin binding protein is expressed in spore mother cell and deposited along with insecticidal protein, Cry1Ac. It interacts with Cry1Ac to potentiate its insecticidal activity and facilitate propagation of Bacillus strain in environment by inhibiting growth of certain fungi.

  18. Structural studies of δ-endotoxin Cry 1 C from Bacillus thuringiensis

    International Nuclear Information System (INIS)

    Guimaraes, B.G.; Garratt, R.C.; Oliva, G.; Lemos, M.V.F.; Arantes, O.M.N.

    1996-01-01

    Full text. The δ-endotoxins are a family of crystal protein by a soil bacterium, Bacillus thuringiensis. The study of these proteins has been of great interest due to their highly specific activity against insects of the orders Lepidoptera, Diptera and Coleoptera. Thus, the δa-endotoxins have been used for more than two decades as biological insecticides to control agricultural pests and, more recently, insects vectors of some diseases. The knowledge of their three-dimensional structures is very important to understand their mechanism of action and their high specificity. To date, the structure of only three proteins of the δ-endotoxins family have been reported: Cry3A, a coleopteran-specific toxin (beetle toxin) 1 , Cry1Aa, a lepidopteran-specific toxin (butterfly toxin) 2 and CytB, a dipteran-specific toxin (mosquito toxin) 3 Our work is aimed at the determination of the crystallographic structure by X-ray diffraction of δ-endotoxin Cry1C, also toxic to insects of the Lepidoptera order but towards families other than those affected by Cry1Aa. A comparison between these structures may lead to important conclusions about the reasons for the specificity and would allow the planning of mutants with more efficient activity. The cry1C gene was cloned into an adequate vector and expressed in an acrystalliferous B. thuringiensis strain. After cell culture and sporulation the microcrystals of Cry1C were separated by ultra-centrifugation in sacharose. The protoxin inclusion bodies were activated by commercial trpsin and the protease-resistant core was purified by anion-exchange chromatography. Crystallization experiments are being conducted in order to obtain single crystals suitable for diffraction measurements. We intend to use the Protein Crystallograph Station of the LNLS to collect data as soon as it is available and we have suitable crystals. (author)

  19. Isolation and molecular characterization of Bacillus thuringiensis found in soils of the Cerrado region of Brazil, and their toxicity to Aedes aegypti larvae

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    Katiane dos Santos Lobo

    Full Text Available ABSTRACT This study investigated the potential of Bacillus thuringiensis isolates obtained in the Cerrado region of the Brazilian state of Maranhão for the biological control of Aedes aegypti larvae. The isolates were obtained from soil samples and the identification of the B. thuringiensis colonies was based on morphological characteristics. Bioassays were run to assess the pathogenicity and toxicity of the different strains of the B. thuringiensis against third-instar larvae of A. aegypti. Protein profiles were obtained by sodium dodecyl sulfate polyacrylamide gel electrophoresis. Polymerase chain reaction assays were used to detect the toxin genes found in the bacterial isolates. Overall, 12 (4.0% of the 300 isolates obtained from 45 soil samples were found to present larvicidal activity, with the BtMA-104, BtMA-401 and BtMA-560 isolates causing 100% of mortality. The BtMA-401 isolate was the most virulent, with the lowest median lethal concentration (LC50 (0.004 × 107 spores/mL, followed by the Bacillus thuringiensis var. israelensis standard (0.32 × 107 spores/mL. The protein profiles of BtMA-25 and BtMA-401 isolates indicated the presence of molecular mass consistent with the presence of the proteins Cry4Aa, Cry11Aa and Cyt1, similar to the profile of Bacillus thuringiensis var. israelensis IPS-82. Surprisingly, however, none of the cry and cyt genes analyzed were amplified in the isolate BtMA-401. The results of the present study revealed the larvicidal potential of B. thuringiensis isolates found in the soils of the Cerrado region from Maranhão, although further research will be necessary to better elucidate and describe other genes associated with the production of insecticidal toxins in these isolates.

  20. Proteomic analysis of Bacillus thuringiensis at different growth phases by using an automated online two-dimensional liquid chromatography-tandem mass spectrometry strategy.

    Science.gov (United States)

    Huang, Shaoya; Ding, Xuezhi; Sun, Yunjun; Yang, Qi; Xiao, Xiuqing; Cao, Zhenping; Xia, Liqiu

    2012-08-01

    The proteome of a new Bacillus thuringiensis subsp. kurstaki strain, 4.0718, from the middle vegetative (T(1)), early sporulation (T(2)), and late sporulation (T(3)) phases was analyzed using an integrated liquid chromatography (LC)-based protein identification system. The system comprised two-dimensional (2D) LC coupled with nanoscale electrospray ionization (ESI) tandem mass spectrometry (MS/MS) on a high-resolution hybrid mass spectrometer with an automated data analysis system. After deletion of redundant proteins from the different batches and B. thuringiensis subspecies, 918, 703, and 778 proteins were identified in the respective three phases. Their molecular masses ranged from 4.6 Da to 477.4 Da, and their isoelectric points ranged from 4.01 to 11.84. Function clustering revealed that most of the proteins in the three phases were functional metabolic proteins, followed by proteins participating in cell processes. Small molecular and macromolecular metabolic proteins were further classified according to the Kyoto Encyclopedia of Genes and Genome and BioCyc metabolic pathway database. Three protoxins (Cry2Aa, Cry1Aa, and Cry1Ac) as well as a series of potential intracellular active factors were detected. Many significant proteins related to spore and crystal formation, including sporulation proteins, help proteins, chaperones, and so on, were identified. The expression patterns of two identified proteins, CotJc and glutamine synthetase, were validated by Western blot analysis, which further confirmed the MS results. This study is the first to use shotgun technology to research the proteome of B. thuringiensis. Valuable experimental data are provided regarding the methodology of analyzing the B. thuringiensis proteome (which can be used to produce insecticidal crystal proteins) and have been added to the related protein database.

  1. Toxicity and residual control of Plutella xylostella L. (Lepidoptera: Plutellidae with Bacillus thuringiensis Berliner and insecticides Toxicidade e controle residual de Plutella xylostella L. (Lepidoptera: Plutellidae com Bacillus thuringiensis Berliner e inseticidas

    Directory of Open Access Journals (Sweden)

    Carla Pedroso de Moraes

    2012-08-01

    Full Text Available Plutella xylostella L. is the most important worldwide pest of cruciferous plants and indiscriminate use of insecticides has led to the resistance of the species to different groups. This research was conducted to compare the toxicity and persistence of two strains of Bacillus thuringiensis to P. xylostella larvae. Concentrations between 125 and 500g 100L-1 of water of the commercial products were evaluated and compared to the insect growth inhibitor diflubenzuron and to the neurotoxic pyrethroid deltamethrin. The efficacy of the insecticides was compared between treated plants kept indoor greenhouse and outdoor. Third instar larvae were more susceptible to B. thuringiensis than first instar ones. Agree and Dipel showed similar control rates of third instars until 10 days after treatment, but on the 15th day, Agree was significantly more efficient than Dipel. Both B. thuringiensis formulations showed a reduction in mortality after 10 days when the treated plants were exposed to natural weather conditions in comparison to the same treatments kept inside greenhouse. Dimilin (100g 100L-1 of water and deltamethrin (30ml of commercial product 100L-1 of water were not efficient to control third instar larvae of P. xylostella. This inefficiency cannot be attributed to a resistant population of P. xylostella since the larval population used in the experiments was not subjected to insecticide pressure, once the crop was organically cultivated all year round. The results showed that both formulations of B. thuringiensis are sound alternatives for the control of the diamondback moth in organically conducted cruciferous crops, considering the high residual control provided under subtropical weather conditions.Larvas de Plutella xylostella L. são as principais pragas de crucíferas cultivadas e o uso excessivo e indiscriminado de inseticidas tem levado a resistência da espécie para diferentes grupos de inseticidas. Este trabalho foi conduzido para

  2. Fragments of Tenebrio molitor cadherin enhance Cry3Aa toxicity for the red flour beetle, Tribolium castaneum (Herbst)

    Czech Academy of Sciences Publication Activity Database

    Moustafa, M.A.M.; Vlasák, Josef; Sehnal, František

    2016-01-01

    Roč. 140, č. 4 (2016), s. 277-286 ISSN 0931-2048 Institutional support: RVO:60077344 Keywords : Bacillus thuringiensis Cry3Aa * biocontrol * toxicity modulation Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 1.641, year: 2016

  3. Bombyx mori ABC transporter C2 structures responsible for the receptor function of Bacillus thuringiensis Cry1Aa toxin.

    Science.gov (United States)

    Tanaka, Shiho; Endo, Haruka; Adegawa, Satomi; Iizuka, Ami; Imamura, Kazuhiro; Kikuta, Shingo; Sato, Ryoichi

    2017-12-01

    Because Bombyx mori ABC transporter C2 (BmABCC2) has 1000-fold higher potential than B. mori cadherin-like protein as a receptor for Bacillus thuringiensis Cry1Aa toxin (Tanaka et al., 2013), the gate-opening ability of the latent pore under six extracellular loops (ECLs) of BmABCC2 was expected to be the reason for its higher potential (Heckel, 2012). In this study, cell swelling assays in Sf9 cells showed that BmABCC2 mutants lacking substrate-excreting activity retained receptor activity, indicating that the gate-opening activity of BmABCC2 is not responsible for Cry1Aa toxicity. The analysis of 29 BmABCC2 mutants demonstrated that 770 DYWL 773 of ECL 4 comprise a putative binding site to Cry1Aa. This suggests that specific toxicity of Cry1Aa toxin to a restricted range of lepidopteran insects is dependent on conservation and variation in the amino acid residues around 770 DYWL 773 of ECL 4 in the ABCC2. Copyright © 2017 Elsevier Ltd. All rights reserved.

  4. Isolation of bacillus thuringiensis from different samples from Mansehra District

    International Nuclear Information System (INIS)

    Younis, F.; Lodhi, A.F.; Raza, G.

    2009-01-01

    The insecticidal activity of Bacillus thuringiensis has made it very interesting for the control of a variety of agricultural pests and human disease vectors. The present study is an attempt to explore the potential and diversity. of Bacillus thuringiensis. from the local environment for the control of cotton spotted bollworm (Earias sp.), a major pest of cotton. Two hundred and ninety eight samples of soil, grain dust, wild animal dung, birds dropping, decaying leaves and dead insects were collected from different ecological environments of Mansehra District yielding 438 Bacillus thuringiensis isolates that produce parasporal crystalline inclusions. In this study the soil samples were found to be the richest source for Bacillus thuringiensis. (author)

  5. Selection of optimum conditions of medium acidity and aeration for submerget cultivation of Bacillus thuringiensis and Beauveria bassiana

    Directory of Open Access Journals (Sweden)

    O. A. Dregval

    2010-06-01

    Full Text Available The paper deals with the influence of medium pH and aeration rate on growth and sporulation of Bacillus thuringiensis and Вeauveria bassiana, which are main constituents of the complex microbial insecticide. It was established optimal medium pH for B. thuringiensis – 6.0 and for В. bassiana – 6.0–7.0. The maximum productivity of the studied microorganisms was observed in the same range of aeration – 7– 14 mmol O2/l/h. The selected conditions of cultivation are necessary for the production of complex biological insecticide based on the association of B. thuringiensis and B. bassiana.

  6. Contributions of gut bacteria to Bacillus thuringiensis-induced mortality vary across a range of Lepidoptera

    Directory of Open Access Journals (Sweden)

    Holt Jonathan

    2009-03-01

    Full Text Available Abstract Background Gut microbiota contribute to the health of their hosts, and alterations in the composition of this microbiota can lead to disease. Previously, we demonstrated that indigenous gut bacteria were required for the insecticidal toxin of Bacillus thuringiensis to kill the gypsy moth, Lymantria dispar. B. thuringiensis and its associated insecticidal toxins are commonly used for the control of lepidopteran pests. A variety of factors associated with the insect host, B. thuringiensis strain, and environment affect the wide range of susceptibilities among Lepidoptera, but the interaction of gut bacteria with these factors is not understood. To assess the contribution of gut bacteria to B. thuringiensis susceptibility across a range of Lepidoptera we examined larval mortality of six species in the presence and absence of their indigenous gut bacteria. We then assessed the effect of feeding an enteric bacterium isolated from L. dispar on larval mortality following ingestion of B. thuringiensis toxin. Results Oral administration of antibiotics reduced larval mortality due to B. thuringiensis in five of six species tested. These included Vanessa cardui (L., Manduca sexta (L., Pieris rapae (L. and Heliothis virescens (F. treated with a formulation composed of B. thuringiensis cells and toxins (DiPel, and Lymantria dispar (L. treated with a cell-free formulation of B. thuringiensis toxin (MVPII. Antibiotics eliminated populations of gut bacteria below detectable levels in each of the insects, with the exception of H. virescens, which did not have detectable gut bacteria prior to treatment. Oral administration of the Gram-negative Enterobacter sp. NAB3, an indigenous gut resident of L. dispar, restored larval mortality in all four of the species in which antibiotics both reduced susceptibility to B. thuringiensis and eliminated gut bacteria, but not in H. virescens. In contrast, ingestion of B. thuringiensis toxin (MVPII following antibiotic

  7. Cry64Ba and Cry64Ca, Two ETX/MTX2-Type Bacillus thuringiensis Insecticidal Proteins Active against Hemipteran Pests.

    Science.gov (United States)

    Liu, Yonglei; Wang, Yinglong; Shu, Changlong; Lin, Kejian; Song, Fuping; Bravo, Alejandra; Soberón, Mario; Zhang, Jie

    2018-02-01

    Genetically modified crops that express insecticidal Bacillus thuringiensis (Bt) proteins have become a primary approach for control of lepidopteran (moth) and coleopteran (beetle) pests that feed by chewing the plants. However, the sap-sucking insects (Hemiptera) are not particularly susceptible to Bt toxins. In this study, we describe two Cry toxins (Cry64Ba and Cry64Ca) from Bt strain 1012 that showed toxicity against two important hemipteran rice pests, Laodelphax striatellus and Sogatella furcifera Both of these proteins contain an ETX/MTX2 domain and share common sequence features with the β-pore-forming toxins. Coexpression of cry64Ba and cry64Ca genes in the acrystalliferous Bt strain HD73 - resulted in high insecticidal activity against both hemipteran pests. No toxicity was observed on other pests such as Ostrinia furnacalis , Plutella xylostella , or Colaphellus bowringi Also, no hemolytic activity or toxicity against cancer cells was detected. Binding assays showed specific binding of the Cry64Ba/Cry64Ca toxin complex to brush border membrane vesicles isolated from L. striatellus Cry64Ba and Cry64Ca are Bt Cry toxins highly effective against hemipteran pests and could provide a novel strategy for the environmentally friendly biological control of rice planthoppers in transgenic plants. IMPORTANCE In Asia, rice is an important staple food, whose production is threatened by rice planthoppers. To date, no effective Bacillus thuringiensis (Bt) protein has been shown to have activity against rice planthoppers. We cloned two Bt toxin genes from Bt strain 1012 that showed toxicity against small brown planthoppers ( Laodelphax striatellus ) and white-backed planthoppers ( Sogatella furcifera ). To our knowledge, the proteins encoded by the cry64Ba and cry64Ca genes are the most efficient insecticidal Bt Cry proteins with activity against hemipteran insects reported so far. Cry64Ba and Cry64Ca showed no toxicity against some lepidopteran or coleopteran pests

  8. Activity of a Brazilian strain of Bacillus thuringiensis israelensis against the cotton Boll Weevil Anthonomus grandis Boheman (Coleoptera: Tenebrionidae).

    Science.gov (United States)

    Monnerat, R; Martins, E; Praça, L; Dumas, V; Berry, C

    2012-02-01

    A Brazilian Bacillus thuringiensis subspecies israelensis, toxic to Diptera, including mosquitoes, was found also to show toxicity to the coleopteran boll weevil Anthonomus grandis Boheman at an equivalent level to that of the standard coleopteran-active B. thuringiensis subspecies tenebrionis T08017. Recombinant B. thuringiensis strains expressing the individual Cyt1Aa, Cry4Aa, Cry4Ba and Cry11Aa toxins from this strain were assessed to evaluate their potential contribution to the activity against A. grandis, either alone or in combination. Whilst individual toxins produced mortality, none was sufficiently potent to allow calculation of LC50 values. Combinations of toxins were unable to attain the same potency as the parental B. thuringiensis subsp. israelensis, suggesting a major role for other factors produced by this strain.

  9. Expression of a Synthetic Gene for the Major Cytotoxin (Cyt1Aa of Bacillus thuringiensis subsp. israelensis in the Chloroplast of Wild-Type Chlamydomonas

    Directory of Open Access Journals (Sweden)

    Seongjoon Kang

    2018-05-01

    Full Text Available Chlamydomonas reinhardtii (Chlamydomonas strains that are toxic to mosquito larvae because they express chloroplast transgenes that are based on the mosquitocidal proteins of Bacillus thuringiensis subsp. israelensis (Bti could be very useful in mosquito control. Chlamydomonas has several advantages for this approach, including genetic controls not generally available with industrial algae. The Bti toxin is produced by sporulating bacteria and has been used for mosquito control for >30 years without creating highly resistant mosquito populations. The suite of toxins is four main proteins: three Cry proteins and the cytotoxic Cyt1Aa (27 kDa. Cyt1Aa is not very toxic to mosquitoes by itself, but it prevents the development of resistance. The production of Cyt1Aa in other microbes, however, has been challenging due to its affinity for certain membrane phospholipids. Here we report on the production of recombinant Cyt1Aa (rCyt1A in the chloroplast of photosynthetic Chlamydomonas at levels of at least 0.3% total protein. Live cell bioassays demonstrated toxicity of the rCyt1Aa Chlamydomonas to larvae of Aedes aegypti. We also expressed the chloroplast cyt1Aa gene in a wild-type Chlamydomonas strain (21 gr that can grow on nitrate. These results have implications for developing a Chlamydomonas strain that will be toxic to mosquito larvae but will not induce strongly resistant populations.

  10. Ecological aspects of Bacillus thuringiensis in an Oxisol Ecologia do Bacillus thuringiensis num Latossolo

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    Lessandra Heck Paes Leme Ferreira

    2003-02-01

    Full Text Available Bacillus thuringiensis is a Gram positive, sporangial bacterium, known for its insecticidal habilities. Survival and conjugation ability of B. thuringiensis strains were investigated; vegetative cells were evaluated in non-sterile soil. Vegetative cells decreased rapidly in number, and after 48 hours the population was predominantly spores. No plasmid transfer was observed in non-sterile soil, probably because the cells died and the remaining cells sporulated quickly. Soil is not a favorable environment for B. thuringiensis multiplication and conjugation. The fate of purified B. thuringiensis toxin was analyzed by extractable toxin quantification using ELISA. The extractable toxin probably declined due to binding on surface-active particles in the soil.O comportamento de células vegetativas do Bacillus thuringiensis foi estudado em solo não esterilizado. Após o inóculo grande parte das células morrem e o restante esporula em 24 horas. Não foi observada conjugação provavelmente porque poucas células sobrevivem no solo e rapidamente esporulam, mostrando que este não é o ambiente propício para a multiplicação e conjugação desta bactéria. A toxina purificada, portanto livre de células, diminui rapidamente sua quantidade em solo não esterilizado. Provavelmente a ligação da toxina na fração argilosa do solo é a principal responsável por este fenômeno.

  11. Correspondence of High Levels of Beta-Exotoxin I and the Presence of cry1B in Bacillus thuringiensis

    Science.gov (United States)

    Espinasse, Sylvain; Gohar, Michel; Chaufaux, Josette; Buisson, Christophe; Perchat, Stéphane; Sanchis, Vincent

    2002-01-01

    Examination of 640 natural isolates of Bacillus thuringiensis showed that the 58 strains (9%) whose supernatants were toxic to Anthonomus grandis (Coleoptera: Curculionidae) produced between 10 and 175 μg of β-exotoxin I per ml. We also found that 55 (46%) of a sample of 118 strains whose culture supernatants were not toxic to A. grandis nevertheless produced between 2 and 5 μg/ml. However, these amounts of β-exotoxin I were below the threshold for detectable toxicity against this insect species. Secretion of large amounts of β-exotoxin I was strongly associated with the presence of cry1B and vip2 genes in the 640 natural B. thuringiensis isolates studied. We concluded that strains carrying cry1B and vip2 genes also possess, on the same plasmid, genetic determinants necessary to promote high levels of production of β-exotoxin I. PMID:12200263

  12. Histopathological Effects of Bt and TcdA Insecticidal Proteins on the Midgut Epithelium of Western Corn Rootworm Larvae (Diabrotica virgifera virgifera

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    Andrew J. Bowling

    2017-05-01

    Full Text Available Western corn rootworm (WCR, Diabrotica virgifera virgifera LeConte is a major corn pest in the United States, causing annual losses of over $1 billion. One approach to protect against crop loss by this insect is the use of transgenic corn hybrids expressing one or more crystal (Cry proteins derived from Bacillus thuringiensis. Cry34Ab1 and Cry35Ab1 together comprise a binary insecticidal toxin with specific activity against WCR. These proteins have been developed as insect resistance traits in commercialized corn hybrids resistant to WCR feeding damage. Cry34/35Ab1 is a pore forming toxin, but the specific effects of Cry34/35Ab1 on WCR cells and tissues have not been well characterized microscopically, and the overall histopathology is poorly understood. Using high-resolution resin-based histopathology methods, the effects of Cry34/35Ab1 as well as Cry3Aa1, Cry6Aa1, and the Photorhabdus toxin complex protein TcdA have been directly visualized and documented. Clear symptoms of intoxication were observed for all insecticidal proteins tested, including swelling and sloughing of enterocytes, constriction of midgut circular muscles, stem cell activation, and obstruction of the midgut lumen. These data demonstrate the effects of these insecticidal proteins on WCR midgut cells, and the collective response of the midgut to intoxication. Taken together, these results advance our understanding of the insect cell biology and pathology of these insecticidal proteins, which should further the field of insect resistance traits and corn rootworm management.

  13. Proteomic Analysis of Bacillus thuringiensis Strain 4.0718 at Different Growth Phases

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    Xiaohui Li

    2012-01-01

    Full Text Available The growth process of Bacillus thuringiensis Bt4.0718 strain was studied using proteomic technologies. The proteins of Bt whole cells at three phases—middle vegetative, early sporulation, and late sporulation—were extracted with lysis buffer, followed with separation by 2-DE and identified by MALDI-TOF/TOF MS. Bioactive factors such as insecticidal crystal proteins (ICPs including Cry1Ac(3, Cry2Aa, and BTRX28, immune inhibitor (InhA, and InhA precursor were identified. InhA started to express at the middle vegetative phase, suggesting its contribution to the survival of Bt in the host body. At the early sporulation phase, ICPs started their expression. CotJC, OppA, ORF1, and SpoIVA related to the formation of crystals and spores were identified, the expression characteristics of which ensured the stable formation of crystals and spores. This study provides an important foundation for further exploration of the stable expression of ICPs, the smooth formation of crystals, and the construction of recombinant strains.

  14. Development of ELISA for the detection of transgenic vegetative insecticidal protein in GM crops/produce.

    Science.gov (United States)

    Kumar, R

    2012-01-11

    In the process of the development of insect-resistant genetically modified (GM) crops and also to evaluate the consistency in the expression of toxin under field conditions, immunological assays are commonly being used. An immunoassay was developed to support the labelling of vegetative insecticidal protein (Vip3A)-based GM produce. The developed ELISA for the measurement of Vip3A is a triple antibody sandwich procedure utilising a polyclonal capture antibody (mouse anti-Vip3A) and a polyclonal detection antibody (rabbit anti-Vip3A) followed by use of a third HRP-conjugated anti-species antibody (goat anti-rabbit IgG). The limit of detection limit of the ELISA assay was 16 ng ml(-1) with a linear quantification range from approximately 31 to 500 ng ml(-1) of Vip3A protein. Furthermore, the assay was in-house validated with GM brinjal samples. The assay was specific, sensitive and reproducible, which can be helpful to detect and track down the spread of unapproved and intentionally/unintentionally released GM produce harbouring Vip protein.

  15. Cytotoxicity analysis of three Bacillus thuringiensis subsp. israelensis δ-endotoxins towards insect and mammalian cells.

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    Roberto Franco Teixeira Corrêa

    Full Text Available Three members of the δ-endotoxin group of toxins expressed by Bacillus thuringiensis subsp. israelensis, Cyt2Ba, Cry4Aa and Cry11A, were individually expressed in recombinant acrystalliferous B. thuringiensis strains for in vitro evaluation of their toxic activities against insect and mammalian cell lines. Both Cry4Aa and Cry11A toxins, activated with either trypsin or Spodoptera frugiperda gastric juice (GJ, resulted in different cleavage patterns for the activated toxins as seen by SDS-PAGE. The GJ-processed proteins were not cytotoxic to insect cell cultures. On the other hand, the combination of the trypsin-activated Cry4Aa and Cry11A toxins yielded the highest levels of cytotoxicity to all insect cells tested. The combination of activated Cyt2Ba and Cry11A also showed higher toxic activity than that of toxins activated individually. When activated Cry4Aa, Cry11A and Cyt2Ba were used simultaneously in the same assay a decrease in toxic activity was observed in all insect cells tested. No toxic effect was observed for the trypsin-activated Cry toxins in mammalian cells, but activated Cyt2Ba was toxic to human breast cancer cells (MCF-7 when tested at 20 µg/mL.

  16. NFATc3 and VIP in Idiopathic Pulmonary Fibrosis and Chronic Obstructive Pulmonary Disease.

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    Anthony M Szema

    Full Text Available Idiopathic pulmonary fibrosis (IPF and chronic obstructive pulmonary disease (COPD are both debilitating lung diseases which can lead to hypoxemia and pulmonary hypertension (PH. Nuclear Factor of Activated T-cells (NFAT is a transcription factor implicated in the etiology of vascular remodeling in hypoxic PH. We have previously shown that mice lacking the ability to generate Vasoactive Intestinal Peptide (VIP develop spontaneous PH, pulmonary arterial remodeling and lung inflammation. Inhibition of NFAT attenuated PH in these mice suggesting a connection between NFAT and VIP. To test the hypotheses that: 1 VIP inhibits NFAT isoform c3 (NFATc3 activity in pulmonary vascular smooth muscle cells; 2 lung NFATc3 activation is associated with disease severity in IPF and COPD patients, and 3 VIP and NFATc3 expression correlate in lung tissue from IPF and COPD patients. NFAT activity was determined in isolated pulmonary arteries from NFAT-luciferase reporter mice. The % of nuclei with NFAT nuclear accumulation was determined in primary human pulmonary artery smooth muscle cell (PASMC cultures; in lung airway epithelia and smooth muscle and pulmonary endothelia and smooth muscle from IPF and COPD patients; and in PASMC from mouse lung sections by fluorescence microscopy. Both NFAT and VIP mRNA levels were measured in lungs from IPF and COPD patients. Empirical strategies applied to test hypotheses regarding VIP, NFATc3 expression and activity, and disease type and severity. This study shows a significant negative correlation between NFAT isoform c3 protein expression levels in PASMC, activity of NFATc3 in pulmonary endothelial cells, expression and activity of NFATc3 in bronchial epithelial cells and lung function in IPF patients, supporting the concept that NFATc3 is activated in the early stages of IPF. We further show that there is a significant positive correlation between NFATc3 mRNA expression and VIP RNA expression only in lungs from IPF patients

  17. Identification and cloning of two insecticidal protein genes from ...

    African Journals Online (AJOL)

    Bacillus thuringiensis (Bt) is the most widely applied type of microbial pesticide due to its high specificity and environmental safety. The activity of Bt is largely attributed to the insecticidal crystal protein encoded by the cry genes. Different insecticidal crystal proteins of Bt have different bioactivity against distinct agricultural ...

  18. Molecular Approaches to Improve the Insecticidal Activity of Bacillus thuringiensis Cry Toxins

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    Wagner A. Lucena

    2014-08-01

    Full Text Available Bacillus thuringiensis (Bt is a gram-positive spore-forming soil bacterium that is distributed worldwide. Originally recognized as a pathogen of the silkworm, several strains were found on epizootic events in insect pests. In the 1960s, Bt began to be successfully used to control insect pests in agriculture, particularly because of its specificity, which reflects directly on their lack of cytotoxicity to human health, non-target organisms and the environment. Since the introduction of transgenic plants expressing Bt genes in the mid-1980s, numerous methodologies have been used to search for and improve toxins derived from native Bt strains. These improvements directly influence the increase in productivity and the decreased use of chemical insecticides on Bt-crops. Recently, DNA shuffling and in silico evaluations are emerging as promising tools for the development and exploration of mutant Bt toxins with enhanced activity against target insect pests. In this report, we describe natural and in vitro evolution of Cry toxins, as well as their relevance in the mechanism of action for insect control. Moreover, the use of DNA shuffling to improve two Bt toxins will be discussed together with in silico analyses of the generated mutations to evaluate their potential effect on protein structure and cytotoxicity.

  19. Profile of cry from native Bacillus thuringiensis isolates and ...

    African Journals Online (AJOL)

    The characterization of 255 Bacillus thuringiensis isolates of Coorg, Sharavatti and BR hills, containing genes known to be active against coleopteran and lepidopteran insect species was done through PCR amplification using the specific and degenerate primers. The isolates were also tested for their insecticidal activity ...

  20. Bt Proteins Have No Detrimental Effects on Larvae of the Green Lacewing, Chrysopa pallens (Rambur) (Neuroptera: Chrysopidae).

    Science.gov (United States)

    Ali, I; Zhang, S; Muhammad, M S; Iqbal, M; Cui, J-J J-J

    2018-06-01

    Biosafety of a genetically modified crop is required to be assessed prior to its commercialization. For this, a suitable artificial diet was developed and used to establish a dietary exposure test for assessing the toxicity of midgut-active Bt insecticidal proteins on Chrysopa pallens (Rambur). Subsequently, this dietary exposure test was used to evaluate the toxicity of the proteins Cry1Ab, Cry1Ac, Cry1Ah, Cry1Ca, Cry1F, Cry2Aa, Cry2Ab, and Vip3Aa on C. pallens larvae. Temporal stability, bioactivity, and the intake of the insecticidal proteins were confirmed by enzyme-linked immunosorbent assay and a sensitive-insect bioassay. The life history characteristics, such as survival, pupation, adult emergence, 7-day larval weight, larval developmental time, and emerged male and female fresh weights remained unaffected, when C. pallens were fed the pure artificial diet (negative control) and the artificial diets containing 200 μg/g of each purified protein: Cry1Ab, Cry1Ac, Cry1Ah, Cry1Ca, Cry1F, Cry2Aa, Cry2Ab, or Vip3Aa. On the contrary, all of the life history characteristics of C. pallens larvae were adversely affected when fed artificial diet containing boric acid (positive control). The results demonstrate that diets containing the tested concentrations of Cry1Ab, Cry1Ac, Cry1Ah, Cry1Ca, Cry1F, Cry2Aa, Cry2Ab, and Vip3Aa have null effects on C. pallens larvae. The outcome indicates that genetically modified crops expressing the tested Bt proteins are safe for the lacewing, C. pallens.

  1. The occurrence of Photorhabdus-like toxin complexes in Bacillus thuringiensis

    Science.gov (United States)

    Recently, genomic sequencing of a Bacillus thuringiensis (Bt) isolate from our collection revealed the presence of an apparent operon encoding an insecticidal toxin complex (Tca) similar to that first described from the entomopathogen Photorhabdus luminescens. To determine whether these genes are w...

  2. Potato expressing beetle-specific Bacillus thuringiensis Cry3Aa toxin reduces performance of a moth

    Czech Academy of Sciences Publication Activity Database

    Hussein, H. M.; Habuštová, Oxana; Turanli, Ferit; Sehnal, František

    2006-01-01

    Roč. 32, č. 1 (2006), s. 1-13 ISSN 0098-0331 R&D Projects: GA ČR(CZ) GA522/02/1507 Institutional research plan: CEZ:AV0Z50070508 Keywords : Bacillus thuringiensis * Spodoptera littoralis * Leptinotarsa decemlineata Subject RIV: GF - Plant Pathology, Vermin, Weed, Plant Protection Impact factor: 1.896, year: 2006

  3. Frequency of Cry1F Non-Recessive Resistance Alleles in North Carolina Field Populations of Spodoptera frugiperda (Lepidoptera: Noctuidae).

    Science.gov (United States)

    Li, Guoping; Reisig, Dominic; Miao, Jin; Gould, Fred; Huang, Fangneng; Feng, Hongqiang

    2016-01-01

    Fall armyworm, Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae), is a target species of transgenic corn (Zea mays L.) that expresses single and pyramided Bacillus thuringiensis (Bt) toxin. In 2014, S. frugiperda were collected from a light trap in North Carolina, and a total of 212 F1/F2 isofemale lines of S. frugiperda were screened for resistance to Bt and non-Bt corn. All of the 212 isolines were susceptible to corn tissue expressing Cry1A.105 + Cry2Ab, Cry1F + Cry1A.105 + Cry2Ab, and Cry1F + Cry1Ab + Vip3Aa20. Growth rate bioassays were performed to isolate non-recessive Bt resistance alleles. Seven individuals out of the 212 isofemale lines carried major non-recessive alleles conferring resistance to Cry1F. A pooled colony was created from the seven individuals. This colony was 151.21 times more resistant to Cry1F than a known-susceptible population and was also resistant to Cry1A.105, but was not resistant to Cry2Ab and Vip3Aa20. The results demonstrate that field populations of S. frugiperda collected from North Carolina are generally susceptible to Cry1F, but that some individuals carry resistant alleles. The data generated in this study can be used as baseline data for resistance monitoring.

  4. Bacillus thuringiensis subsp. israelensis and Its Dipteran-Specific Toxins

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    Eitan Ben-Dov

    2014-03-01

    Full Text Available Bacillus thuringiensis subsp. israelensis (Bti is the first Bacillus thuringiensis to be found and used as an effective biological control agent against larvae of many mosquito and black fly species around the world. Its larvicidal activity resides in four major (of 134, 128, 72 and 27 kDa and at least two minor (of 78 and 29 kDa polypeptides encoded respectively by cry4Aa, cry4Ba, cry11Aa, cyt1Aa, cry10Aa and cyt2Ba, all mapped on the 128 kb plasmid known as pBtoxis. These six δ-endotoxins form a complex parasporal crystalline body with remarkably high, specific and different toxicities to Aedes, Culex and Anopheles larvae. Cry toxins are composed of three domains (perforating domain I and receptor binding II and III and create cation-selective channels, whereas Cyts are composed of one domain that acts as well as a detergent-like membrane perforator. Despite the low toxicities of Cyt1Aa and Cyt2Ba alone against exposed larvae, they are highly synergistic with the Cry toxins and hence their combinations prevent emergence of resistance in the targets. The lack of significant levels of resistance in field mosquito populations treated for decades with Bti-bioinsecticide suggests that this bacterium will be an effective biocontrol agent for years to come.

  5. Insecticide susceptibility of Aedes albopictus and Ae. aegypti from Brazil and the Swiss-Italian border region.

    Science.gov (United States)

    Suter, Tobias; Crespo, Mônica Maria; de Oliveira, Mariana Francelino; de Oliveira, Thaynan Sama Alves; de Melo-Santos, Maria Alice Varjal; de Oliveira, Cláudia Maria Fontes; Ayres, Constância Flávia Junqueira; Barbosa, Rosângela Maria Rodrigues; Araújo, Ana Paula; Regis, Lêda Narcisa; Flacio, Eleonora; Engeler, Lukas; Müller, Pie; Silva-Filha, Maria Helena Neves Lobo

    2017-09-19

    Aedes aegypti and Ae. albopictus are two highly invasive mosquito species, both vectors of several viruses, including dengue, chikungunya and Zika. While Ae. aegypti is the primary vector in the tropics and sub-tropics, Ae. albopictus is increasingly under the public health watch as it has been implicated in arbovirus-transmission in more temperate regions, including continental Europe. Vector control using insecticides is the pillar of most control programmes; hence development of insecticide resistance is of great concern. As part of a Brazilian-Swiss Joint Research Programme we set out to assess whether there are any signs of existing or incipient insecticide resistance primarily against the larvicide Bacillus thuringiensis svar. israelensis (Bti), but also against currently applied and potentially alternative insecticides in our areas, Recife (Brazil) and the Swiss-Italian border region. Following World Health Organization guidelines, dose-response curves for a range of insecticides were established for both colonized and field caught Ae. aegypti and Ae. albopictus. The larvicides included Bti, two of its toxins, Cry11Aa and Cry4Ba, Lysinibacillus sphaericus, Vectomax CG®, a formulated combination of Bti and L. sphaericus, and diflubenzuron. In addition to the larvicides, the Swiss-Italian Ae. albopictus populations were also tested against five adulticides (bendiocarb, dichlorodiphenyltrichloroethane, malathion, permethrin and λ-cyhalothrin). Showing a similar dose-response, all mosquito populations were fully susceptible to the larvicides tested and, in particular, to Bti which is currently used both in Brazil and Switzerland. In addition, there were no signs of incipient resistance against Bti as larvae were equally susceptible to the individual toxins, Cry11Aa and Cry4Ba. The field-caught Swiss-Italian populations were susceptible to the adulticides tested but DDT mortality rates showed signs of reduced susceptibility. The insecticides currently used for

  6. Radioimmunoassay of vasoactive intestinal polypeptide (VIP) in plasma. [/sup 125/I tracer techniques

    Energy Technology Data Exchange (ETDEWEB)

    Fahrenkrug, J.; Schaffalitzky de Muckadell, O.B.

    1977-06-01

    A sensitive and specific radioimmunoassay for vasoactive intestinal polypeptide (VIP) has been developed, which can detect 3.3 pmol x L/sup -1/ of the peptide in plasma. Antisera to highly purified porcine VIP coupled to albumin were raised in eight rabbits. The final dilution, the avidity, and the specificity of each antiserum were determined. /sup 125/I-VIP served as label, and highly purified porcine VIP was used as standard. Separation of antibody-bound and free VIP was achieved by plasma-coated charcoal. Nonspecific interference with the assay system was excluded by extraction of plasm samples with ethanol. The reliability of the assay was investigated by recovery experiments, by serial dilution of plasma samples with high concentration of endogenous VIP, and by immunosorption. The within-and between assay reproducibility at a concentration of 18.3 pmol x L/sup -1/ was 1.6 and 2.3 pmol x L/sup -1/ (1 S.D.), respectively. Median fasting concentration of VIP in plasma from 74 normal subjects was 7.3 pmol x L/sup -1/ (range: 0 to 20.0 pmol x L/sup -1/).

  7. Modification of Cry4Aa toward Improved Toxin Processing in the Gut of the Pea Aphid, Acyrthosiphon pisum.

    Directory of Open Access Journals (Sweden)

    Michael A Rausch

    Full Text Available Aphids are sap-sucking insects (order: Hemiptera that cause extensive damage to a wide range of agricultural crops. Our goal was to optimize a naturally occurring insecticidal crystalline (Cry toxins produced by the soil-dwelling bacterium Bacillus thuringiensis for use against the pea aphid, Acyrthosiphon pisum. On the basis that activation of the Cry4Aa toxin is a rate-limiting factor contributing to the relatively low aphicidal activity of this toxin, we introduced cathepsin L and cathepsin B cleavage sites into Cry4Aa for rapid activation in the aphid gut environment. Incubation of modified Cry4Aa and aphid proteases in vitro demonstrated enhanced processing of the toxin into the active form for some of the modified constructs relative to non-modified Cry4Aa. Aphids fed artificial diet with toxin at a final concentration of 125 μg/ml showed enhanced mortality after two days for one of the four modified constructs. Although only modest toxin improvement was achieved by use of this strategy, such specific toxin modifications designed to overcome factors that limit aphid toxicity could be applied toward managing aphid populations via transgenic plant resistance.

  8. VIP receptors from porcine liver: High yield solubilization in a GTP-insensitive form

    International Nuclear Information System (INIS)

    Voisin, T.; Couvineau, A.; Guijarro, L.; Laburthe, M.

    1990-01-01

    Vasoactive intestinal peptide (VIP) receptors were solubilized from porcine liver membranes using CHAPS. The binding of 125 I-VIP to solubilized receptors was reversible, saturable and specific. Scatchard analysis indicated the presence of one binding site with a Kd of 6.5 ± 0.3 nM and a Bmax of 1.20 ± 0.15 pmol/mg protein. Solubilized and membrane-bound receptors displayed the same pharmacological profile since VIP and VIP-related peptides inhibited 125 I-VIP binding to both receptor preparations with the same rank order of potency e.g. VIP>helodermin>rat GRF>rat PHI>secretin>human GRF. GTP inhibited 125 I-VIP binding to membrane-bound receptors but not to solubilized receptors supporting functional uncoupling of VIP receptor and G protein during solubilization. Affinity labeling of solubilized and membrane-bound VIP receptors with 125 I-VIP revealed the presence of a single molecular component with Mr 55,000 in both cases. It is concluded that VIP receptors from porcine liver can be solubilized with a good yield, in a GTP-insensitive, G protein-free form. This represents a major advance towards the purification of VIP receptors

  9. Fatores de virulência de Bacillus thuringiensis: o que existe além das proteínas Cry

    Directory of Open Access Journals (Sweden)

    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.

  10. Detection of the mosquitocidal toxin genes encoding Cry11 proteins from Bacillus thuringiensis using a novel PCR-RFLP method Detección de genes que codifican proteínas mosquitocidas Cry11 de Bacillus thuringiensis mediante un método de PCR-RFLP novedoso

    Directory of Open Access Journals (Sweden)

    D. H. Sauka

    2010-02-01

    Full Text Available A polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP method for detection of cry11 genes from Bacillus thuringiensis was established. Based on the analysis of conserved regions of the cry11 genes, 2 oligonucleotide primers were designed to amplify a 1459-bp fragment of the cry11Aa gene, and a 1471-bp of the cry11Ba and cry11Bb genes. The amplification products were digested with restriction endonuclease HinfI. Exotic B. thuringiensis strains and native isolates collected from soils, leaves and stored product dust of Argentina were analyzed to study the distribution of cry11 genes. The PCR-RFLP patterns revealed the detection of cry11 genes in 3 of 64 exotic strains and in 10 of 107 native B. thuringiensis isolates tested. Just the cry11Aa gene subclass was detected among these bacteria. Since the methodology was also developed to detect cry11Ba and cry11Bb genes, an experimental future confirmation will be required. Based on the results obtained, the PCR-RFLP method presented may be a valuable tool for specific detection of the mosquitocidal toxin genes encoding Cry11 proteins from B. thuringiensis.En el presente estudio se estableció una estrategia basada en la amplificación génica (PCR y el posterior análisis de restricción (RFLP para detectar todos los genes cry11 de Bacillus thuringiensis informados hasta ahora. De acuerdo con el análisis de las regiones conservadas en los genes cry11, se diseñaron dos cebadores para amplificar un fragmento de 1459 pb de los genes cry11Aa y un fragmento de 1471 pb de los genes cry11Ba y cry11Bb. Los productos de la amplificación fueron digeridos con la enzima de restricción HinfI. Se analizaron cepas exóticas de B. thuringiensis y aislamientos nativos de Argentina obtenidos a partir de muestras de suelos, hojas y polvillo de silos, para estudiar la distribución de los genes cry11. Los patrones de PCR-RFLP revelaron la presencia de genes cry11 en 3 de las 64 cepas ex

  11. IMPACT OF BT ( BACILLUS THURINGIENSIS ) CROPS ON BAT ACTIVITY IN SOUTH TEXAS AGROECOSYSTEMS

    Science.gov (United States)

    The widespread adoption of transgenic insecticidal crops raises concerns that nontarget species may be harmed and food webs disrupted. The goal of this research is to determine how transgenic Bt (Bacillus thuringiensis) crops impact the activity of Brazilian freetailed bats (Tada...

  12. Microimaging of Bacillus thuringiensis Toxin-binding proteins in gypsy moth larval gut using confocal fluorescence microscopy

    Science.gov (United States)

    Daniel J. Krofcheck; Algimantas P. Valaitis

    2010-01-01

    After ingestion by susceptible insect larvae, Bacillus thuringiensis (Bt) insecticidal proteins bind to the brush border membranes of gut epithelial cells and disrupt the integrity of the plasma membrane by forming...

  13. Characterization of Bacillus thuringiensis soil isolates from Cuba, with insecticidal activity against mosquitoes

    Directory of Open Access Journals (Sweden)

    Aileen González

    2011-09-01

    Full Text Available Chemical insecticides may be toxic and cause environmental degradation. Consequently, biological control for insects represents an alternative with low ecological impact. In this work, three soil isolates (A21, A51 and C17 from different regions of the Cuban archipelago were identified, characterized and evaluated against Aedes aegypti and Culex quinquefasciatus. The new isolates were compared with reference IPS82 strain and two strains isolated from biolarvicides Bactivec and Bactoculicida, respectively. The differentiation was done by morphological, biochemical, bioassays activity and molecular methods (SDS-PAGE, plasmid profile and random amplified polymorphic analysis. All isolates were identified as Bacillus thuringiensis. The A21, A51 and C17 isolates showed higher larvicide activity than Bactivec’s isolated reference strain, against both A. aegypti and C. quinquefasciatus. A21 isolate had a protein profile similar to IPS82 and Bactivec strain. A51 and C17 isolates produced a characteristic proteins pattern. A21 and A51 isolates had plasmid patterns similar to IPS82 standard strain, while C17 isolate had different both plasmid profile and protein bands. All the studied isolates showed a diverse RAPD patterns and were different from the strains previously used in biological control in Cuba. Rev. Biol. Trop. 59 (3: 1007-1016. Epub 2011 September 01.El uso prolongado de insecticidas ha conducido al desarrollo de resistencia en diferentes especies de mosquitos y al incremento de la degradación del ambiente. El control biológico de insectos ha devenido como una alternativa útil y de bajo impacto ambiental. En nuestro estudio fueron identificados, caracterizados tres aislamientos de suelos procedentes de diferentes regiones del archipiélago cubano y comparados con cepas de referencia: aisladas de los biolarvicidas Bactivec y Bactoculicida, además de IPS82. La diferenciación de los mismos se llevó a cabo mediante métodos morfol

  14. Bacillus thuringiensis: the legacy to the XXI century

    Directory of Open Access Journals (Sweden)

    M. Realpe

    1998-01-01

    Full Text Available Bacillus thuringiensis-based insecticides are the main production line of the biopesticides world market. The research devoted to this area, promoted by the necessity to solve problems in agriculture and public health has resulted in an exhaustive knowledge of its biology. The diversity of the B. thuringiensis strains has permitted to develop several products mainly, but not exclusively, for insect control. With the new developments in the field of molecular biology, it has been possible to understand the molecular basis of the mode of action and to increase the range of activity as well. As a result of the broad use in several countries, resistant strains of some of the susceptible insects have appeared. The aim of this review is to elaborate a theoretical framework of the current state of research on B. thuringiensis, describing briefly the knowledge on this bacterium, with emphasis on biological phenomena that underlie its toxic activity and the problems that will be faced during the XXI century with the increasingly common resistance, all this analyzed from a biotechnological perspective.

  15. Synergistic effect of certain insecticides combined with Bacillus thuringiensis on mosquito larvae

    Directory of Open Access Journals (Sweden)

    C.P. Narkhede

    2017-04-01

    Full Text Available For effective vector control it is essential to formulate new preparations having multiple action against the vector pest. Developing combined formulation of biopesticide and chemical pesticide is one of the novel concept to fight against the vectors with new weapons; however, compatibility of biopesticide i.e. Bacillus thuringiensis (Bt and chemical pesticide is a real hurdle. In this investigation, local isolate Bacillus thuringiensis SV2 (BtSV2 was tested for its compatibility with various available mosquito larvicides. Temephos was most compatible with BtSV2 than with other tested pesticides. These two compatible agents were tested for larvicidal potential. Our study revealed that the synergistic effect of both agents reduces LC50 value by 30.68 and 22.36% against the Ae. aegypti and An. stephensi, respectively. The larvicidal potential increased when compared to individual pesticides. It was also observed a biochemical change in larvae after the TBT (Temephos + Bacillus thuringiensis combination treatment; it involves decreased level of alpha esterase, acetylcholine esterase and protein while level of beta esterase and acid phosphatase was unchanged and alkaline phosphatase activity was increased. Increased potential of combined formulation may be due to altered physiological condition.

  16. Recent advancement on chemical arsenal of Bt toxin and its application in pest management system in agricultural field.

    Science.gov (United States)

    Chattopadhyay, Pritam; Banerjee, Goutam

    2018-04-01

    Bacillus thuringiensis ( Bt ) is a Gram-positive, spore-forming, soil bacterium, which is very popular bio-control agent in agricultural and forestry. In general, B. thuringiensis secretes an array of insecticidal proteins including toxins produced during vegetative growth phase (such as secreted insecticidal protein, Sip; vegetative insecticidal proteins, Vip), parasporal crystalline δ-endotoxins produced during vegetative stationary phase (such as cytolytic toxin, Cyt; and crystal toxin, Cry), and β-exotoxins. Till date, a wide spectrum of Cry proteins has been reported and most of them belong to three-domain-Cry toxins, Bin-like toxin, and Etx_Mtx2-like toxins. To the best of our knowledge, neither Bt insecticidal toxins are exclusive to Bt nor all the strains of Bt are capable of producing insecticidal Bt toxins. The lacuna in their latest classification has also been discussed. In this review, the updated information regarding the insecticidal Bt toxins and their different mode of actions were summarized. Before applying the Bt toxins on agricultural field, the non-specific effects of toxins should be investigated. We also have summarized the problem of insect resistance and the strategies to combat with this problem. We strongly believe that this information will help a lot to the budding researchers in the field of modern pest control biotechnology.

  17. Characterization of a Bacillus thuringiensis strain collection isolated from diverse Costa Rican natural ecosystems

    Directory of Open Access Journals (Sweden)

    Glen Arrieta

    2006-03-01

    Full Text Available Costa Rican natural ecosystems are among the most diverse in the world. For this reason, we isolated strains of the entomopathogenic bacteria Bacillus thuringiensis (Bt to determine their diversity, distribution and abundance. A total of 146 Bt strains were obtained from environmental samples collected from diverse natural ecosystems and life zones of Costa Rica. We recovered Bt strains from 71%, 63%, 61% and 54% of soil samples, fresh leaves, other substrates and leaf litter respectively. Bt was isolated in 65%of the samples collected in the humid tropical forest in national parks (Braulio Carrillo, Gandoca Manzanillo, Sierpe, Hitoy Cerere, and Cahuita, and in 59% of the samples collected in the dry tropical forest (Parque Nacional Marino las Baulas, Palo Verde and Santa Rosa. In the very humid tropical forest (Tortuguero Bt was isolated in 75% of the samples and in the very humid tropical forest transition perhumid (Cararait was found in 69% of the samples. The strains exhibit a diverse number, size and morphology of parasporal inclusion bodies: irregular (47%,oval (20%, bipyramidal (3%, bipyramidal and cubic (1%, bipyramidal, oval and irregular (5% and bipyramidal, oval and cubic crystals (2%. Strains isolated from Braulio Carrillo, Tortuguero and Cahuita, presented predominantly irregular crystals. On the other hand, more than 60% of the isolates from Térraba-Sierpe and Hitoy-Cerere had medium oval crystals. Strains from Gandoca-Manzanillo, Palo Verde and Carara presented mainly combinations of oval and irregular crystals. Nevertheless, the greatest diversity in crystal morphology was observed in those from Santa Rosa, Llanos del Río Medio Queso and Parque Marino las Baulas. Protein analyses of the crystal-spore preparations showed ä -endotoxin with diverse electrophoretic patterns, with molecular weights in the range of 20 to 160 kDa. Fifty six percent of the strains amplified with the cry2 primer, 54% with vip3, 20% with cry1, 9% with

  18. Avaliação de produtos à base de Bacillus thuringiensis no controle da traça-das-crucíferas Evaluation of insecticides based on Bacillus thuringiensis in the control of the diamondback moth

    Directory of Open Access Journals (Sweden)

    Patrícia T Medeiros

    2006-06-01

    Full Text Available Avaliou-se em dois experimentos a suscetibilidade da traça-das-crucíferas a inseticidas à base de Bacillus thuringiensis em repolho cv. Itiban. O delineamento do primeiro experimento (de julho a setembro/03, em área de plantio comercial em Brazlândia (DF, foi de blocos casualizados, com seis tratamentos e dez repetições; os bioinseticidas utilizados foram B. thuringiensis kurstaki (S1450CO, B. thuringiensis aizawai comercial (Bta e três produtos formulados com as estirpes S1450BB, S811BB, S845BB de B. thuringiensis pertencentes ao Banco de Bacillus spp. da Embrapa Recursos Genéticos e Biotecnologia. No segundo experimento, realizado no campo experimental da Embrapa (DF, de outubro/03 a janeiro/04, o delineamento foi de blocos casualizados, com seis tratamentos e quatro repetições; os inseticidas utilizados foram os mesmos do primeiro experimento, com a adição de Spinosad, e retirado o tratamento S811BB. Os produtos foram aplicados quando foi atingido o nível de dano de seis furos nas quatro folhas centrais do repolho. O Bta comercial foi o produto mais eficaz no primeiro experimento, tendo sido aplicado cinco vezes e diferiu estaticamente dos demais produtos. Os formulados S845BB e S1450BB não apresentaram diferenças quando comparados ao produto comercial S1450 e foram aplicados seis vezes. O produto S811BB também foi aplicado seis vezes, mas sua eficácia foi inferior aos demais produtos e não diferiu da testemunha. Já no segundo experimento, o S1450 comercial foi aplicado cinco vezes e os demais produtos à base de Bt, seis vezes. Todos os produtos utilizados não diferiram entre si, diferindo apenas no número de aplicações.Two experiments were performed to evaluate the susceptibility of the diamondback moth to insecticides based on Bacillus thuringiensis in cabbage cv. Itiban. The first experiment was carried out from July to September 2003, in a production area in Brazlândia (DF, Brazil. Randomized blocks with six

  19. Acute, sublethal, and combination effects of azadirachtin and Bacillus thuringiensis on the cotton bollworm, Helicoverpa armigera.

    Science.gov (United States)

    Abedi, Zahra; Saber, Moosa; Vojoudi, Samad; Mahdavi, Vahid; Parsaeyan, Ehsan

    2014-02-26

    The cotton bollworm, Helicoverpa armigera Hübner (Lepidoptera: Noctuidae) is a polyphagous and cosmopolitan insect pest that causes damage to various plants. In this study, the lethal and sublethal effects of azadirachtin and Bacillus thuringiensis Berliner sub sp . kurstaki (Bacillales: Bacillaceae) were evaluated on third instar H. armigera under laboratory conditions. The LC50 values of azadirachtin and Bt were 12.95 and 96.8 µg a.i./mL, respectively. A total mortality of 56.7% was caused on third instar larvae when LC20 values of the insecticides were applied in combination with each other. The LT50 values of azadirachtin and Bt were 4.8 and 3.6 days, respectively. The results of the sublethal study showed that the application of LC30 value of azadirachtin and Bt reduced the larval and pupal weight and increased larval and pupal duration of H. armigera. The longevity and fecundity of female adults were affected significantly by the insecticides. Female fecundity was reduced by the treatments, respectively. The lowest adult emergence ratio and pupation ratio were observed in the azadirachtin treatment. The results indicated that both insecticides have high potential for controlling of the pest. This is an open access paper. We use the Creative Commons Attribution 3.0 license that permits unrestricted use, provided that the paper is properly attributed.

  20. Experimental research for tumor VIP receptor imaging

    International Nuclear Information System (INIS)

    Li Qianwei; Tan Tianzhi

    1998-01-01

    To study the possibility of radioactive labelled vasoactive intestinal peptide (VIP) for tumor VIP receptor imaging. 125 I-VIP was prepared by chloramine-T method, and purified by Sephadex G-50 column chromatography. The bioactivity and stability of 125 I-VIP were measured by silica 60 F 254 TLC and competition test to SGC7901 cell in vitro. The biodistribution of 125 I-VIP was studied in the nude mice bearing tumor. The results showed that labelled rate of 125 I was 73.8%, the specific activity was 18.2 PBq/mol, the radiochemical purity (RCP) was over 98% and remained 96.3% after 48 days stored at -80 degree C. The specific binding of 125 I-VIP to the SGC7901 cell was inhibited by VIP in dose dependence in the competition experiment. The radioactivity of tumor was higher than that of muscles in all phases (P<0.05-0.01), the peak activity of tumor occurred at 30 min (3.58 +- 0.48ID%/g) and the peak ratio of T/N occurred at 60 min after the injection. The activity of lungs was obviously higher than that of blood, the intestine was always in low level. Most of the activity in the body was mainly eliminated from kidney. The present study demonstrated that the radioactive labelled VIP is a promising agent for tumor VIP receptor scintigraphy

  1. Occurrence of natural Bacillus thuringiensis contaminants and residues of Bacillus thuringiensis-based insecticides on fresh fruits and vegetables

    DEFF Research Database (Denmark)

    Frederiksen, Kristine; Rosenquist, Hanne; Jørgensen, Kirsten

    2006-01-01

    . kurstaki HD1 present in the products Dipel, Biobit, and Foray, and nine isolates grouped with B. thuringiensis subsp. aizawai present in Turex. The commercial strains were primarily isolated from samples of tomatoes, cucumbers, and peppers. A multiplex PCR method was developed to simultaneously detect all...

  2. Insecticide toxicity to Trichogramma pretiosum (Hymenoptera: Trichogrammatidae) females and effect on descendant generation.

    Science.gov (United States)

    Vianna, Ulysses R; Pratissoli, Dirceu; Zanuncio, José C; Lima, Eraldo R; Brunner, Jay; Pereira, Fabrício F; Serrão, José E

    2009-02-01

    The effect of nine insecticides used in tomato production was evaluated on adults of two Trichogramma pretiosum Riley (Hymenoptera: Trichogrammatidae) populations from Rive and Afonso Cláudio, Espírito Santo State, Brazil. The experiment was developed in an acclimatized chamber at 25 +/- 1 degrees C, 70 +/- 10% relative humidity and 14 h photophase. Eggs of Anagasta kuehniella (Lepidoptera: Pyralidae), previously immersed in insecticides solutions were offered to females of both T. pretiosum populations. Bacillus thuringiensis, lufenuron and triflumuron had lowest negative effects on parasitism and viability of individuals of these populations; however, abamectin and pyrethroids (betacyflurin 50 and 125 g/l and esfenvalerate) insecticides reduced parasitism rates. T. pretiosum emerged from A. kuehniella eggs treated with esfenvalerate but were not able to parasitize non treated eggs of this host. B. thuringiensis, lufenuron and triflumuron may be used in integrated pest management programs to control tomato pests, because they have moderated negative effect on parasitoid wasps.

  3. Beetle-specific Bacillus thuringiensis Cry3Aa toxin reduces larval growth and curbs reproduction in Spodoptera littoralis (Boisd.)

    Czech Academy of Sciences Publication Activity Database

    Hussein, Hany; Habuštová, Oxana; Sehnal, František

    2005-01-01

    Roč. 61, - (2005), s. 1186-1192 ISSN 1526-498X R&D Projects: GA ČR(CZ) GA522/02/1507 Institutional research plan: CEZ:AV0Z50070508 Keywords : Bacillus thuringiensis * Spodoptera littoralis * Bt applications Subject RIV: ED - Physiology Impact factor: 1.175, year: 2005

  4. Estirpes de Bacillus thuringiensis efetivas contra insetos das ordens Lepidoptera, Coleoptera e Diptera Bacillus thuringiensis strains effective against insects of Lepidoptera, Coleoptera and Diptera orders

    Directory of Open Access Journals (Sweden)

    Lílian Botelho Praça

    2004-01-01

    Full Text Available O objetivo deste trabalho foi selecionar entre 300 estirpes de Bacillus thuringiensis as efetivas simultaneamente contra larvas de Spodoptera frugiperda J.E. Smith e Anticarsia gemmatalis Hübner (Lepidoptera: Noctuidae, Anthonomus grandis Boheman (Coleoptera: Curculionidae, Aedes aegypti Linnaeus e Culex quinquefasciatus Say (Diptera: Culicidae. Foram selecionadas duas estirpes de B. thuringiensis, denominadas S234 e S997, que apresentaram atividade contra as três ordens de insetos. As estirpes foram caracterizadas por métodos morfológicos, bioquímicos e moleculares. As mesmas apresentaram duas proteínas principais de 130 e 65 kDa, produtos de reação em cadeia da polimerase de tamanho esperado para a detecção dos genes cry1Aa, cry1Ab, cry1Ac, cry1B e cry2 e cristais bipiramidais, cubóides e esféricos.The aim of this work was to select among 300 strains of Bacillus thuringiensis those which are simultaneously effective against larvae of Spodoptera frugiperda J.E. Smith and Anticarsia gemmatalis Hübner (Lepidoptera: Noctuidae, Anthonomus grandis Boheman (Coleoptera: Curculionidae, Aedes aegypti Linnaeus and Culex quinquefasciatus Say (Diptera: Culicidae. Two strains of B. thuringiensis were selected, S234 and S997, which presented activity against those three insect orders. Both strains were characterized by morphological, biochemical and molecular methods. They have presented two main proteins with 130 and 65 kDa, polimerase chain reaction products with expected sizes for detection of the genes cry1Aa, cry1Ab, cry1Ac, cry1B and cry2 and bipiramidal, cubical and spherical crystals.

  5. Midgut microbiota and host immunocompetence underlie Bacillus thuringiensis killing mechanism

    Science.gov (United States)

    Caccia, Silvia; Di Lelio, Ilaria; La Storia, Antonietta; Marinelli, Adriana; Varricchio, Paola; Franzetti, Eleonora; Banyuls, Núria; Tettamanti, Gianluca; Casartelli, Morena; Giordana, Barbara; Ferré, Juan; Gigliotti, Silvia; Pennacchio, Francesco

    2016-01-01

    Bacillus thuringiensis is a widely used bacterial entomopathogen producing insecticidal toxins, some of which are expressed in insect-resistant transgenic crops. Surprisingly, the killing mechanism of B. thuringiensis remains controversial. In particular, the importance of the septicemia induced by the host midgut microbiota is still debated as a result of the lack of experimental evidence obtained without drastic manipulation of the midgut and its content. Here this key issue is addressed by RNAi-mediated silencing of an immune gene in a lepidopteran host Spodoptera littoralis, leaving the midgut microbiota unaltered. The resulting cellular immunosuppression was characterized by a reduced nodulation response, which was associated with a significant enhancement of host larvae mortality triggered by B. thuringiensis and a Cry toxin. This was determined by an uncontrolled proliferation of midgut bacteria, after entering the body cavity through toxin-induced epithelial lesions. Consequently, the hemolymphatic microbiota dramatically changed upon treatment with Cry1Ca toxin, showing a remarkable predominance of Serratia and Clostridium species, which switched from asymptomatic gut symbionts to hemocoelic pathogens. These experimental results demonstrate the important contribution of host enteric flora in B. thuringiensis-killing activity and provide a sound foundation for developing new insect control strategies aimed at enhancing the impact of biocontrol agents by reducing the immunocompetence of the host. PMID:27506800

  6. Slow release formulations of Bacillus thuringiensis israelensis (AM 65-52 and spinosyns: effectiveness against the West Nile vector Culex pipiens in Saudi Arabia

    Directory of Open Access Journals (Sweden)

    Alaa Sulaiman Alsobhi

    2016-07-01

    Full Text Available Objective: To investigate the effectiveness of slow release formulations of Bacillus thuringiensis israelensis (AM 65-52 (B. thuringiensis israelensis and spinosyns against the West Nile vector Culex pipiens (Cx. pipiens in Saudi Arabia. Methods: We tested slow release insecticide formulations of Natular DT, Tap 60 and VectoBac granule against II instars of Cx. pipiens larvae in 50 L laboratory arenas. Results: Slow release formulations of B. thuringiensis israelensis and spinosyns gave continuous control against Cx. pipiens for several weeks. Natular DT was more effective over Tap 60 and VectoBac granule of about 1.3 and 5.8 times, respectively. Variations in the durations of effective control among the tested slow release formulations may reflect differences in their active ingredients and the mode of action. Conclusions: Our results highlighted the effectiveness of B. thuringiensis israelensis and spinosyns against an important West Nile vector, providing baseline data to develop ecofriendly mosquito control programs in Saudi Arabia.

  7. VIP and its homologous increase vascular conductance in certain endocrine and exocrine glands

    International Nuclear Information System (INIS)

    Huffman, L.J.; Connors, J.M.; Hedge, G.A.

    1988-01-01

    The effects of vasoactive intestinal peptide (VIP) and related structural homologues on tissue vascular conductances were investigated in anesthetized male rats. VIP, peptide histidine isoleucine (PHI), secretin, growth hormone-releasing factor (GHRF), gastric inhibitory peptide (GIP), or saline was infused intravenously over 4 min. Tissue blood flows were measured during this time by use of 141 Ce-labeled microspheres. Circulating thyrotropin (TSH), triiodothyronine (T 3 ), and thyroxine (T 4 ) levels were determined before and at 20 min and 2 h after treatment. Marked increases in thyroid, pancreatic, and salivary gland vascular Cs occurred during peptide infusion with the order of potency correlating with the degree of structural homology to VIP. PHI and secretin produced maximal increases in vascular Cs, which were the same as those obtained with VIP. Circulating TSH, T 3 , and T 4 levels were not different from values in saline-infused rats after peptide treatments that caused striking increases in thyroid vascular C. These observations indicate that the vascular beds of certain endocrine and exocrine glands are responsive to the vasodilatory action of VIP and related homologues with the order of potency corresponding to the degree of structural homology to VIP. These results are also consistent with the proposal that structural homologues of VIP act at the same vascular receptor as VIP. Alternative, the involvement of different vascular receptors, acting through the same mechanism at a level beyond the receptor site, cannot be excluded

  8. Comparative biodistribution profile of [131I]VIP and [131I]VIP10-28

    Directory of Open Access Journals (Sweden)

    Maria Tereza Colturato

    2005-10-01

    Full Text Available Various tumor cells express significantly higher amounts of VIP receptors (VIPR that provided the basis for the clinical use of radiolabeled VIP for the in vivo localization of tumors. This work studied the labeling of VIP and VIP10-28 with iodine-131 to compare the biological distribution of the labeled compounds in Nuce mice and the affinity for tumor cells. Both VIP and VIP10-28 peptides contain two tyrosine residues, in positions 10 and 22, that are theoretically equally susceptible to radioiodination employing oxidative electrophilic substitution using oxidizing agents like Chloramine T. Radiochemical purity of the reaction mixture was determined by electrophoresis and HPLC. The VIP peptide and the fragment were labeled with radioiodine with good radiochemical yield (above 96%. Suitable, but important differences can be observed in biological distribution studies. Comparatively, blood clearance was faster for labeled VIP and perhaps because of this, the uptake in tumor was lower, especially during the first hour. These differences observed in the biological distribution of the compounds can be related to the lipophilicity of the labeled compounds.Várias células tumorais expressam significantemente uma alta quantidade de receptores VIP (VIPR que determinam a base para o uso clínico de VIP radiomarcado para localização de tumores in vivo. Foi estudado neste trabalho a marcação do VIP e do fragmento VIP10-28 com iodo-131 comparando a distribuição biológica dos compostos marcados em camundongos Nude e sua afinidade pelas células tumorais. Ambos os peptídeos, VIP e VIP10-28. contém dois resíduos de tirosina nas posições 10 e 22, que teoricamente são igualmente susceptíveis pela substituição eletrofílica oxidativa do radioiodo utilizando Cloramina T como agente oxidante. A pureza radioquímica da mistura de reação foi determinada por eletroforese e cromatografia líquida de alta eficiência (CLAE. O VIP e fragmento foram

  9. Treatment of an Aedes aegypti colony with the Cry11Aa toxin for 54 generations results in the development of resistance

    Directory of Open Access Journals (Sweden)

    Gloria Cadavid-Restrepo

    2012-02-01

    Full Text Available To study the potential for the emergence of resistance in Aedes aegypti populations, a wild colony was subjected to selective pressure with Cry11Aa, one of four endotoxins that compose the Bacillus thuringiensis serovar israelensis toxin. This bacterium is the base component of the most important biopesticide used in the control of mosquitoes worldwide. After 54 generations of selection, significant resistance levels were observed. At the beginning of the selection experiment, the half lethal concentration was 26.3 ng/mL and had risen to 345.6 ng/mL by generation 54. The highest rate of resistance, 13.1, was detected in the 54th generation. Because digestive proteases play a key role in the processing and activation of B. thuringiensis toxin, we analysed the involvement of insect gut proteases in resistance to the Cry11Aa B. thuringiensis serovar israelensis toxin. The protease activity from larval gut extracts from the Cry11Aa resistant population was lower than that of the B. thuringiensisserovar israelensis susceptible colony. We suggest that differences in protoxin proteolysis could contribute to the resistance of this Ae. aegypti colony.

  10. Bacillus thuringiensis Cry1Ca-resistant Spodoptera exigua lacks expression of one of four Aminopeptidase N genes

    NARCIS (Netherlands)

    Herrero, S.; Gechev, T.; Bakker, P.L.; Moar, W.; Maagd, de R.A.

    2005-01-01

    BACKGROUND: Insecticidal toxins from Bacillus thuringiensis bind to receptors on midgut epithelial cells of susceptible insect larvae. Aminopeptidases N (APNs) from several insect species have been shown to be putative receptors for these toxins. Here we report the cloning and expression analysis of

  11. Avaliação da eficiência de formulações de Bacillus thuringiensis para o controle de traça-das-crucíferas em repolho no Distrito Federal Efficiency of Bacillus thuringiensis formulations in controlling Diamondback Moth in cabbage in the Federal District

    Directory of Open Access Journals (Sweden)

    Marina Castelo Branco

    1999-11-01

    Full Text Available A traça-das-crucíferas (Plutella xylostella é a praga mais importante do repolho no Distrito Federal. Seu controle é feito basicamente com inseticidas. Novos produtos são constantemente avaliados para o controle da praga e neste trabalho duas novas formulações de Bacillus thuringiensis [Bacillus thuringiensis var. aizawai (350 e 500 g/hae B. thuringiensis var. kurstaki x B. thuringiensis var. aizawai (350 e 500 ml/ha] foram avaliadas no período de maio a outubro de 1995. Os tratamentos B. thuringiensis var. kurstaki (500 ml/ha, deltametrina (240 ml/ha e uma testemunha sem pulverização foram também incluídos no experimento. O delineamento foi blocos ao acaso, com sete tratamentos e quatro repetições. Os resultados mostraram que B. thuringiensis var. aizawai nas duas dosagens avaliadas e B. thuringiensis var. kurstaki x B. thuringiensis var. aizawai (500 ml/ha foram os produtos mais eficientes. Ao final do experimento larvas e pupas de traça-das-crucíferas foram coletadas no campo e a primeira geração foi submetida a um teste de laboratório onde discos de folhas de repolho foram tratados com as dosagens dos inseticidas a base de B. thuringiensis utilizadas no campo. Larvas de segundo estádio foram colocadas sobre os discos tratados e a mortalidade de larvas avaliada após 72 h. Todos os tratamentos causaram mais de 97% de mortalidade de larvas. O resultado do teste de laboratório sugere que a menor eficiência de B. thuringiensis var. kurstaki x B. thuringiensis var. aizawai (350 ml/ha e B. thuringiensis var. kurstaki (500 ml/ha no teste de campo quando comparada aos demais Bacillus, pode ser devido à mais rápida degradação destes produtos no ambiente.The Diamondback Moth (Plutella xylostella is the most important cabbage pest in the Federal District. New insecticides are frequently tested for its control and in this study two new Bacillus thuringiensis formulations [B. thuringiensis var. aizawai (350 and 500 g/ha and B

  12. F2 screen, inheritance and cross-resistance of field-derived Vip3A resistance in Spodoptera frugiperda (Lepidoptera: Noctuidae) collected from Louisiana, USA.

    Science.gov (United States)

    Yang, Fei; Morsello, Shannon; Head, Graham P; Sansone, Chris; Huang, Fangneng; Gilreath, Ryan T; Kerns, David L

    2017-11-28

    Fall armyworm, Spodoptera frugiperda, is a target pest of the Vip3A protein used in pyramided Bt corn and cotton in the USA. In this study, we provide the first documentation of a resistance allele conferring Vip3A resistance in a field-derived population of S. frugiperda from the USA, and characterize its inheritance and cross-resistance. An F 2 screen with 104 two-parent families generated from a field collection of S. frugiperda in Louisiana, USA, resulted in one family carrying a Vip3A resistance allele. The Vip3A-resistant strain (RR) derived from the two-parent family showed a high level of resistance to Vip3A in both diet and whole-plant bioassays, with a resistance ratio of >632.0-fold relative to a susceptible population (SS) based on diet-overlay bioassays. The inheritance of Vip3A resistance was monogenic, autosomal and recessive. Furthermore, the Vip3A resistance conferred no cross-resistance to Cry1F, Cry2Ab2 or Cry2Ae purified proteins, with resistance ratios of 3.5, 5.0 and 1.1, respectively. These findings provide valuable information for characterizing Vip3A resistance, resistance monitoring, and developing effective resistance management strategies for the sustainable use of the Vip3A technology. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

  13. Cry1A(b)16 toxin from Bacillus thuringiensis: Theoretical refinement of three-dimensional structure and prediction of peptides as molecular markers for detection of genetically modified organisms.

    Science.gov (United States)

    Plácido, Alexandra; Coelho, Andreia; Abreu Nascimento, Lucas; Gomes Vasconcelos, Andreanne; Fátima Barroso, Maria; Ramos-Jesus, Joilson; Costa, Vladimir; das Chagas Alves Lima, Francisco; Delerue-Matos, Cristina; Martins Ramos, Ricardo; Marani, Mariela M; Roberto de Souza de Almeida Leite, José

    2017-07-01

    Transgenic maize produced by the insertion of the Cry transgene into its genome became the second most cultivated crop worldwide. Cry gene from Bacillus thuringiensis kurstaki expresses protein derivatives of crystalline endotoxins which confer insect resistance onto the maize crop. Mandatory labeling of processed food containing or made by genetically modified organisms is in force in many countries, so, it is very urgent to develop fast and practical methods for GMO identification, for example, biosensors. In the absence of an available empirical structure of Cry1A(b)16 protein, a theoretical model was effectively generated, in this work, by homology modeling and molecular dynamics simulations based on two available homologous protein structures. Molecular dynamics simulations were carried out to refine the selected model, and an analysis of its global structure was performed. The refined models of Cry1A(b)16 showed a standard fold and structural characteristics similar to those seen in Bacillus thuringiensis Cry1A(a) insecticidal toxin and Bacillus thuringiensis serovar kurstaki Cry1A(c) toxin. After in silico analysis of Cry1A(b)16, two immunoreactive candidate peptides were selected and specific polyclonal antibodies were produced resulting in antibody-peptide interaction. Biosensing devices are expected to be developed for detection of the Cry1A(b) protein as a marker of transgenic maize in food. Proteins 2017; 85:1248-1257. © 2017 Wiley Periodicals, Inc. © 2017 Wiley Periodicals, Inc.

  14. Radioiodination of vasoactive intestinal peptide (VIP)

    International Nuclear Information System (INIS)

    Wang, Y.; Wang, L.; Yin, D.

    2002-01-01

    In recent years, increasing biochemical and radiochemical research has been performed to develop radiolabelled peptides as specific ligands for tumour associated receptors. VIP, a 28-amino acid peptide containing two tyrosines and three lysines, has demonstrated that various tumour cells express significantly higher amounts of VIP-receptors and could be applied to the clinic diagnosis. For these purposes, radiohalogenation of VIP by direct and indirect method was studied. Direct labelling works well for radioiodine but is limited to dehalogenation of labelling products in vivo. Conjugate labelling methods including Boltonhunter and wood reagents were developed but introduction of such a molecule to peptides may lead to the decrease of biological activity in vivo. In order to resolve these problems, N-Succinimidyl-3-(tri-nbutylstannyl) benzoate (ATE) was elected for the radioiodination of VIP and already employed to radioiodination of IgG successfully. The in vitro stability and biological activity would be compared in these two methods. Vasoactive intestinal peptide (VIP) and human immunoglobulin (IgG) were radioiodinated by direct and indirect methods. Iodogen was employed in direct method and N-Succinimidyl-3-(tri-n-butylstannyl) benzoate (ATE) was applied as a prosthetic group in the conjugation labelling. The subject of our study was optimizing the radiohalogenation of IgG and VIP followed by separation and analysis of reaction products. The advantages and disadvantages were illustrated by comparing the in vitro stability and biological activity in these two methods. Na 123 I was prepared by nuclear reaction of 124 Te(p, 2n) 123 I using cyclone-30. More than 95% of radiochemical purity, more than 95% of radionuclide purity and about 100 mCi/mL of radioactivity concentration were obtained. ATE was supplied by Dr. Pozzi and radioiodinated with iodogen and 96% of labelling efficiency was obtained. The stability of radioactive S 125 IB kept well in dark at 4

  15. A synthetic cryIC gene, encoding a Bacillus thuringiensis δ-endotoxin, confers Spodoptera resistance in alfalfa and tobacco

    NARCIS (Netherlands)

    Strizhov, N.; Keller, M.; Mathur, J.; Koncz-Kaiman, Z.; Bosch, D.; Prudovksy, E.; Schell, J.; Sneh, B.; Koncz, C.; Zilberstein, A.

    1996-01-01

    Spodoptera species, representing widespread polyphagous insect pests, are resistant to Bacillus thuringiensis δ-endotoxins used thus far as insecticides in transgenic plants. Here we describe the chemical synthesis of a cryIC gene by a novel template directed ligation–PCR method. This simple and

  16. Seleção e caracterização de estirpes de Bacillus thuringiensis eficientes contra a Diatraea saccharalis (Lepidoptera: Crambidae Selection and characterization of Bacillus thuringiensis efficient strains against Diatraea saccharalis (Lepidoptera: Crambidae

    Directory of Open Access Journals (Sweden)

    Cristina Lima de Macedo

    2012-12-01

    Full Text Available O objetivo deste trabalho foi selecionar e caracterizar estirpes nativas de Bacillus thuringiensis tóxicas a Diatraea saccharalis (Lepidoptera: Crambidae. Cento e seis estirpes pertencentes ao Banco de Bactérias de Invertebrados, da Embrapa Recursos Genéticos e Biotecnologia, foram testadas quanto à toxicidade a D. saccharalis, e, as mais tóxicas, caracterizadas por métodos bioquímicos e moleculares. Das 106 estirpes testadas, 16 causaram 100% de mortalidade em 24 horas. As três estirpes mais tóxicas apresentaram concentração letal média entre 8 e 43 ng cm-2. O perfil proteico das 16 estirpes mostrou a presença de proteínas de 130 e 65 kDa, e a caracterização molecular mostrou a presença dos genes tipo cry1 e cry2: cry1Aa, cry1Ab, cry1Ac e cry2Aa. A concentração letal média de esporos e cristais obtidos a partir de estirpes recombinantes, que expressavam individualmente os genes cry1Aa, cry1Ab, cry1Ac e cry2Aa, variou entre 222 e 610 ng cm-2, valores muito superiores aos das estirpes nativas mais tóxicas, que apresentavam possibilidade de expressão simultânea desses genes. Este resultado é indicativo de que há sinergia entre as toxinas. Há interação entre as toxinas de B. thuringiensis e seus receptores na broca-do-colmo da cana-de-açúcar.The objective of this work was to select and characterize native strains of Bacillus thuringiensis toxic to Diatraea saccharalis (Lepidoptera: Crambidae. A hundred-and-six strains, belonging to the bank of invertebrate bacteria (Brazil, of Embrapa Genetic Resources and Biotechnology, were tested as to their toxicity to D. saccharalis, and the most toxic ones were characterized by biochemical and molecular methods. Out of the 106 tested strains, 16 caused 100% mortality within 24 hours. The three most toxic strains showed median lethal concentrations between 8 and 43 ng cm-2. The protein profile of the 16 strains showed the presence of 130 and 65 kDa proteins, and the molecular

  17. Morphological and functional correlates of VIP neurons in cerebral cortex

    International Nuclear Information System (INIS)

    Magistretti, P.J.; Morrison, J.H.; Shoemaker, W.J.; Bloom, F.E.

    1984-01-01

    Vasoactive Intestinal Polypeptide (VIP) promotes the hydrolysis of 3H-glycogen newly synthesized from 3H-glucose by mouse cortical slices. This effect occurs rapidly, approximately 50% of the maximal effect being reached within one minute. The maximal effect is achieved after 5 minutes and maintained for at least 25 minutes. Furthermore the glycogenolytic effect of VIP is reversible, and pharmacologically specific. Thus several neuropeptides present in cerebral cortex such as cholecystokinin-8, somatostatin-28, somatostatin-14, met-enkephalin, leu-enkephalin, do not affect 3H-glycogen levels. VIP fragments 6-28, 16-28 and 21-28 are similarly inactive. Furthermore, among the peptides which share structural homologies with VIP, such as glucagon, secretin, PHI-27 and Gastric Inhibitory Peptide, only secretin and PHI-27 promote 3H-glycogen hydrolysis, with EC50 of 500 and 300 nM respectively, compared to an EC50 of 25 nM for VIP. Immunohistochemical observations indicate that each VIP-containing bipolar cell is identified with a unique radical cortical volume, which is generally between 15-60 micrograms in diameter and overlaps with the contiguous domains of neighbouring VIP-containing bipolar cells. Thus this set of biochemical and morphological observations support the notion that VIP neurons have the capacity to regulate the availability of energy substrates in cerebral cortex locally, within circumscribed, contiguous, radial domains

  18. Digestive enzyme as benchmark for insecticide resistance development in Culex pipiens larvae to chemical and bacteriologic insecticides.

    Science.gov (United States)

    Kamel, Nashwa H; Bahgat, Iman M; El Kady, Gamal A

    2013-04-01

    This work monitored changes in some digestive enzymes (trypsin and aminopeptidase) associated with the building up of resistance in Cx. pipiens larvae to two chemical insecticides (methomyl and/or malathion) and one biological insecticide (Bacillus thuringiensis-H14 or B.t H 14). The LC50 value of methomyl for both field- and the 12th generation (F12) of the selected strain was 1.789 ppm and 8.925 ppm respectively. The LC50 value of malathion for both field and the F12 of the selected strain was 0.082 ppm and 0.156 ppm respectively, and those of B.t H14 of field strain and the F12 was 2.550ppm & 2.395ppm respectively. The specific activity of trypsin enzyme in control susceptible colony was 20.806 +/- 0.452micromol/min/mg protein; but at F4 and F8 for malathion and methomyl treated larvae were 10.810 +/- 0.860 & 15.616+/-0.408 micromol/min/mg protein, respectively. Trypsin activity of F12 in treated larvae with B.t.H14 was 2.097 +/- 0.587 microiol/min/mg protein. Aminopeptidase specific activity for susceptible control larvae was 173.05 +/- 1.3111 micromol/min/mg protein. This activity decreased to 145.15 +/- 4.12, 152.497 +/- 6.775 & 102.04 +/- 3.58a micromol/min/mg protein after larval (F 12) treatment with methomyl, malathion and B.t H 14 respectively.

  19. Activity of wild-type and hybrid Bacillus thuringiensis delta-endotoxins against Agrotis ipsilon

    NARCIS (Netherlands)

    Maagd, de R.A.; Weemen-Hendriks, M.; Molthoff, J.W.; Naimov, S.

    2003-01-01

    Twelve Cry1 and two Cry9 ?-endotoxins fromBacillus thuringiensis were tested for their activity against black cutworm (Agrotis ipsilon).A. ipsilon was not susceptible to many toxins, but three toxins had significant activity. Cry9Ca was the most toxic, followed by Cry1Aa and Cry1Fb. Hybrids between

  20. Effects of various insecticides on the development of the egg parasitoid Trichogramma dendrolimi (Hymenoptera: Trichogrammatidae).

    Science.gov (United States)

    Takada, Y; Kawamura, S; Tanaka, T

    2001-12-01

    The toxicity of six insecticides, acephate, methomyl, ethofenprox, cartap, chlorfluazuron, and Bacillus thuringiensis (Bt) was tested on different developmental stages of the egg parasitoid, Trichogramma dendrolimi (Matsumura). Each of the insecticides tested showed different degrees of toxicity to the parasitoid, Ethofenprox showed the highest toxicity and cartap showed relatively higher toxicity compared with the other insecticides. The development of the parasitoids treated with these two insecticides was normal, similar to that of the control group. Only the emergence of adult wasps from host eggs was disturbed. Emergence of the host, Mamestra brassicae larva was reduced following treatment with ethofenprox, cartap and methomyl. However, adult female wasps, which emerged from host eggs treated with the insecticides had the ability to oviposit normally.

  1. Investigation of Cytocidal Activity of Bacillus Thuringiensis Parasporal Toxin on CCRF-CEM Cell Line

    Directory of Open Access Journals (Sweden)

    Elham Moazamian

    2013-03-01

    Full Text Available Background & Objective: Parasporin is a parasporal protein of Bacillus thuringiensis and exhibits special cytocidal activity against human cancer cells. Similar to other insecticidal Bacillus thuringiensis crystal toxins, parasporin shows target specificity and damages the cellular membrane. In this study, different strains of Bacillus thuringiensis isolated from various regions of Iran and their cytocidal activity against CCRF-CEM cell line and human erythrocyte were investigated.   Materials & Methods: Fifty soil samples were collected from different Iranian provinces, and characterization was performed based on protein crystal morphology by phase-contrast microscope and variations of Cry protein toxin using SDS-PAGE. After parasporin was processed with proteinase K, the active form was produced and protein activity on the cell line was evaluated. Results: Parasporal inclusion proteins showed different cytotoxicity against acute lymphoblastic leukemia cells (ALL, but not against normal lymphocyte. Isolated parasporin demonstrated no hemolytic activity against human erythrocyte. It appears that these proteins have the ability to differentiate between normal lymphocytes and leukemia cells and have specific receptors on specific cancer cell lines. Conclusion: Our results provide evidence that the parasporin-producing organism is a common member in Bacillus thuringiensis populations occurring in the natural environments of Iran.

  2. Long lasting persistence of Bacillus thuringiensis Subsp. israelensis (Bti in mosquito natural habitats.

    Directory of Open Access Journals (Sweden)

    Mathieu Tilquin

    Full Text Available BACKGROUND: The detrimental effects of chemical insecticides on the environment and human health have lead to the call for biological alternatives. Today, one of the most promising solutions is the use of spray formulations based on Bacillus thuringiensis subsp. israelensis (Bti in insect control programs. As a result, the amounts of Bti spread in the environment are expected to increase worldwide, whilst the common belief that commercial Bti is easily cleared from the ecosystem has not yet been clearly established. METHODOLOGY/MAIN FINDINGS: In this study, we aimed to determine the nature and origin of the high toxicity toward mosquito larvae found in decaying leaf litter collected in several natural mosquito breeding sites in the Rhône-Alpes region. From the toxic fraction of the leaf litter, we isolated B. cereus-like bacteria that were further characterized as B. thuringiensis subsp. israelensis using PCR amplification of specific toxin genes. Immunological analysis of these Bti strains showed that they belong to the H14 group. We finally used amplified length polymorphism (AFLP markers to show that the strains isolated from the leaf litter were closely related to those present in the commercial insecticide used for field application, and differed from natural worldwide genotypes. CONCLUSIONS/SIGNIFICANCE: Our results raise the issue of the persistence, potential proliferation and environmental accumulation of human-spread Bti in natural mosquito habitats. Such Bti environmental persistence may lengthen the exposure time of insects to this bio-insecticide, thereby increasing the risk of resistance acquisition in target insects, and of a negative impact on non-target insects.

  3. Diversity of Bacillus thuringiensis strains in the maize and bean phylloplane and their respective soils in Colombia.

    Science.gov (United States)

    Jara, S; Maduell, P; Orduz, S

    2006-07-01

    To evaluate the distribution of Bacillus thuringiensis strains from maize and bean phylloplane and their respective soils. B. thuringiensis was isolated from the phylloplane and soil of maize and bean from three municipalities in Antioquia, Colombia. Ninety six samples of phylloplane and 24 of soil were analyzed. A total of 214 isolates were obtained from 96 phylloplane samples while 59 isolates were recovered from 24 soil samples. Sixty five per cent and 12% of the phylloplane and soil isolates, respectively, showed activity against Spodoptera frugiperda. These isolates contained delta-endotoxin proteins of 57 and 130 kDa. The most toxic isolates against S. frugiperda had the genotype cry1Aa, cry1Ac, cry1B, and cry1D. In contrast, 27% of the phylloplane isolates and 88% of the soil isolates were active against Culex quinquefasciatus and had protein profiles similar to B. thuringiensis serovar. medellin and B. thuringiensis serovar. israelensis. The most active isolates contain cry4 and cry11 genes. The predominant population of B. thuringiensis on the phylloplane harbored the cry1 gene and was active against S. frugiperda, whereas in soil, isolates harboring cry11 gene and active against C. quinquefasciatus were the majority. The predominance of specific B. thuringiensis populations, both on the leaves and in the soil, suggests the presence of selection in B. thuringiensis populations on the studied environment.

  4. Further research on the biological function of inclusion bodies of Anomala cuprea entomopoxvirus, with special reference to the effect on the insecticidal activity of a Bacillus thuringiensis formulation.

    Science.gov (United States)

    Mitsuhashi, Wataru; Asano, Shoji; Miyamoto, Kazuhisa; Wada, Sanae

    2014-01-01

    Entomopoxviruses (EVs) form two types of inclusion body: spheroids, which contain virions, and spindles, which do not. The authors tested whether the spindles from a coleopteran EV, Anomala cuprea EV (ACEV), enhanced the insecticidal activity of a commercial Bacillus thuringiensis (Bt) formulation and the susceptibility of scarabaeid pest species in Japan to the virus's spheroids, to assess whether ACEV inclusion bodies are potential biological control agents for pest insects. Peroral inoculation with both ACEV spindles and the Bt toxin only or the complete Bt formulation shortened the survival and increased the mortality of treated insects compared with those of insects inoculated with Bt without the spindles (8-38 h of decrease in LT50 values among assays). ACEV showed high infectivity to a major scarabaeid pest species in Japanese sugar cane fields. The results suggest that spindles or the constituent protein fusolin can be used as a coagent with Bt formulations, and that fusolin coexpression with a Bt toxin in crops might improve the insecticidal efficacy. In addition, the spheroids are potential biocontrol agents for some scarabaeid pests that are not easy to control because of their underground habitation. © 2013 Society of Chemical Industry.

  5. Efektivitas Bacillus thuringiensis dalam Pengendalian Larva Nyamuk Anopheles sp.

    Directory of Open Access Journals (Sweden)

    Citra Inneke Wibowo

    2017-08-01

    Full Text Available Nyamuk Anopheles sp adalah vektor penyakit malaria. Pengendalian vektor penyakit malaria dapat dilakukan secara biologis yaitu dengan menggunakan Bacillus thuringiensis. Tujuan penelitian adalah untuk mengetahui efektivitas konsentrasi Bacillus thuringiensis dalam pengendalian larva nyamuk Anopheles sp.Penelitian ini dilakukan secara eksperimental menggunakan Rancangan Acak Lengkap Faktorial (RAL Faktorial yang terdiri atas dua faktor yaitu konsentrasi Bacillus thuringiensis dan stadia larva Anopheles dengan pengulangan tiga kali.Perlakuan yang dicobakan adalahkonsentrasi Bacillus thuringiensis (A yang terdiri atas 5 taraf:A0: konsentrasi B.thuringiensis 0 CFU.mL-1, A1: konsentrasi B.thuringiensis 102 CFU.mL-1, A2: konsentrasi B.thuringiensis 104 CFU.mL-1, A3: konsentrasi B.thuringiensis 106CFU.mL-1, A4: konsentrasi B.thuringiensis 108CFU.mL-1. Perlakuan tahapan instar larva Anopheles sp. (B adalah sebagai berikut:B1: stadia larva instar I, B2: stadia larva instar II, B3: stadia larva instar III, B4: stadia larva instar IVsehingga terdapat 60 satuan percobaan. Hasil penelitian  menunjukkan konsentrasi B. thuringiensis isolat CK dan IPB CC yang paling berpengaruh dalam pengendalian larva Anopheles sp adalah 108 CFU.mL-1 . Instar larva yang paling peka terhadap B. thuringiensis isolat IPB CC adalah instar I dan II sedangkan instar yang peka terhadap isolat CK adalah instar II, Perlakuan konsentrasi isolat B. thuringiensis dan tingkat instar larva yang paling baik dalam pengendalian larva Anopheles sp. adalah 108 CFU.mL-1, dan instar I dan II.

  6. Localization of Bacillus thuringiensis Cry1A toxin-binding molecules in gypsy moth larval gut sections using fluorescence microscopy

    Science.gov (United States)

    Algimantas P. Valaitis

    2011-01-01

    The microbial insecticide Bacillus thuringiensis (Bt) produces Cry toxins, proteins that bind to the brush border membranes of gut epithelial cells of insects that ingest it, disrupting the integrity of the membranes, and leading to cell lysis and insect death. In gypsy moth, Lymantria dispar, two toxin-binding molecules for the...

  7. Cytotoxic effects of delfin insecticide (Bacillus thuringiensis) on cell ...

    African Journals Online (AJOL)

    STORAGESEVER

    2008-08-04

    Aug 4, 2008 ... In acute exposure cells showed deformities such as swelling of cells, oval shaped deformity, and ... Commercial grade of delfin insecticide used in this study was manufactured by .... exposure to cigarette extracts. Antibiotics caused .... administration of a neem pesticide on rat metabolic enzymes. J. Environ.

  8. Elucidation of the mechanisms of CryIIIA overproduction in a mutagenized strain of Bacillus thuringiensis var. tenebrionis

    International Nuclear Information System (INIS)

    Adams, L.F.; Mathewes, S.; O'Hara, P.; Peterson, A.; Gürtler, H.

    1994-01-01

    NB176 is a Bacillus thuringiensis mutant derived by λ-irradiation of NB125 Bacillus thuringiensis var. tenebrionis (Krieg). It exhibits two interesting phenotypes: (i) oligosporogeny and (ii) twofold to threefold overproduction of the CryIIIA protein. Southern profiles of the NB176 strain showed an additional copy(s) of the cryIIIA gene located on a 4 kb HindIII fragment, in addition to the expected cryIIIA gene on a 3 kb HindIII fragment. Each cryIIIA gene-bearing HindIII fragment was cloned from NB176. The restriction map of the 3 kb HindIII fragment was identical to that published by Donovan and coworkers. Sequencing of the 4 kb HindIII fragment showed no alterations in the promoter region of the cryIIIA gene but did show replacement of the region immediately following the cryIIIA open reading frame with a sequence encoding a transposase with 50% amino acid homology to that of Tn 1000. These findings suggest that the overproduction phenotype of NB176 results from extra copies of the cryIIIA gene produced from a transposition event(s) induced or stabilized by γ-irradiation. Integration of additional copies of the cryIIIA gene into the native 90MDa plasmid of the wild-type B. thuringiensis var. tenebrionis strain resulted in strains that made enormous crystals, many possessing greatly enhanced insecticidal activity

  9. Baseline sensitivity of maize borers in India to the Bacillus thuringiensis insecticidal proteins Cry1A.105 and Cry2Ab2.

    Science.gov (United States)

    Jalali, Sushil K; Yadavalli, Lalitha; Ojha, Rakshit; Kumar, Pradyumn; Sulaikhabeevi, Suby B; Sharma, Reema; Nair, Rupa; Kadanur, Ravi C; Kamath, Subray P; Komarlingam, Mohan S

    2015-08-01

    Among the major pests of maize in India are two stem borers, Chilo partellus (Swinhoe) and Sesamia inferens (Walker), and an earworm, Helicoverpa armigera (Hübner). As a pest control strategy, transgenic Bacillus thuringiensis (Bt) maize hybrids are undergoing regulatory trials in India. We have determined the sensitivity of the target lepidopterans to the insecticidal Bt proteins expressed in Bt maize, as this determines product efficacy and the resistance management strategy to be adopted. Maize hybrids with event MON89034 express two insecticidal Bt proteins, Cry1A.105 and Cry2Ab2. Sensitivity profiles of 53 populations of C. partellus, 21 populations of S. inferens and 21 populations of H. armigera, collected between 2008 and 2013 from maize-growing areas in India, to Cry1A.105 and Cry2Ab2 proteins were generated through dose-response assays. Cry1A.105 protein was the most effective to neonates of C. partellus (mean MIC90 range 0.30-1.0 µg mL(-1) ) and H. armigera (mean MIC90 range 0.71-8.22 µg mL(-1) ), whereas Cry2Ab2 (mean MIC90 range 0.65-1.70 µg mL(-1) ) was the most effective to S. inferens. Populations of C. partellus, S. inferens and H. armigera were susceptible to the Bt proteins Cry1A.105 and Cry2Ab2. The Bt sensitivity data will serve as precommercialisation benchmarks for resistance monitoring purposes. © 2014 Society of Chemical Industry.

  10. Antitumor effects of radioiodinated antisense oligonucleotide mediated by VIP receptor

    International Nuclear Information System (INIS)

    Ou Xiaohong; Tan Tianzhi; Li Yunchun; Kuang Anren

    2004-01-01

    Purpose: we had constructed a targeting delivery system based on intestinal peptide (VIP) for antisense oligonucleotide (ASON) transfer into VIP receptor-positive cells in previous study. The aims of present studies are to observe the antitumor effect of VIP-131I-ASON in HT29 human colon adenocarcinoma xenografts. Methods: A 15-met phosphorothioate ASON, which was complementary to the translation start region of the C-myc oncogene mRNA, was labeled with 131I and the labelled compound was linked to the VIP bound covalently 'to a polylysine chain so as to deliver oligonucleotide into tumor cells. Distribution experiments for evaluating the radiolabeled antisense complexe uptake in tumor tissue were performed in BALB/c nude mice bearing with HT29 tumor xenografts. Nude mice beating HT29 tumor xenografts were adminstered VIP-131I-ASON (3.7,7.4 MBq) or 131I-ASON (3.7 MBq), 131I labeled control sense and nosense DNA (3.7 MBq), or saline. Antitumor effects were assessed using endpoints of tumor growth delay. C-myc-encoded protein expression of tumor was measured by immunocytohistochemical staining. Results: Distribution experiment performed with athymic mice bearing human colon tumor xenografts revealed maximal accumulation of conjugated ASON in the tumor tissue 2 h after administration and significantly higher than that in nude mice injected unconjngated ASON [(5.89±1.03)%ID/g and(1.56±0.31)%ID/g, respectively; t=7.7954 P<0.001]. The radioratio of tumor to muscle was peaked 4h after administration. VIP-131I-ASON exhibited strong antitumor effects against HT29 xenografts, decreasing their growth rate 7-fold compare with that in saline-treated mice(tumor growth delay, 25.4±0.89 day). The antitumor effects of unconjugated 131I-ASON were much less profound than VIP-131I-ASON (tumor growth delay, 3.2±1.3 and 25.4±0.89 day, respectively; q=51.4126 P<0.01). Sense, nosense control ON with VIP carder caused no therapeutic effect. There was no progressive weight loss or

  11. [Susceptibility of Aedes aegypti (L.) strains from Havana to a Bacillus thuringiensis var. israelensis].

    Science.gov (United States)

    Menéndez Díaz, Zulema; Rodríguez Rodríguez, Jinnay; Gato Armas, René; Companioni Ibañez, Ariamys; Díaz Pérez, Manuel; Bruzón Aguila, Rosa Yirian

    2012-01-01

    the integration of chemical and biological methods is one of the strategies for the vector control, due to the existing environmental problems and the concerns of the community as a result of the synthetic organic insecticide actions. The bacterium called Bacillus thuringiensis var. israelensis in liquid formulation has been widely used in the vector control programs in several countries and has shown high efficacy at lab in Cuba. to determine the susceptibility of Aedes aegypti collected in the municipalities of La Habana province to Bacillus thuringiensis var. israelensis. fifteen Aedes aegypti strains, one from each municipality, were used including larvae and pupas collected in 2010 and one reference strain known as Rockefeller. The aqueous formulation of Bacillus thuringiensis var. israelensis (Bactivec, Labiofam, Cuba) was used. The bioassays complied with the World Health Organization guidelines for use of bacterial larvicides in the public health sector. The larval mortality was read after 24 hours and the results were processed by the statistical system SPSS (11.0) through Probit analysis. the evaluated mosquito strains showed high susceptibility to biolarvicide, there were no significant differences in LC50 values of Ae. aegypti strains, neither in the comparison of these values with those of the reference strain. the presented results indicate that the use of Bacillus thuringiensis var. israelensis continues to be a choice for the control of Aedes aegypti larval populations in La Habana province.

  12. Baseline sensitivity of lepidopteran corn pests in India to Cry1Ab insecticidal protein of Bacillus thuringiensis.

    Science.gov (United States)

    Jalali, Sushil K; Lalitha, Yadavalli; Kamath, Subray P; Mohan, Komarlingam S; Head, Graham P

    2010-08-01

    Genetically engineered corn (Bt corn) expressing Bacillus thuringiensis Berliner insecticidal protein Cry1Ab is a biotechnological option being considered for management of lepidopteran corn pests in India. As a resistance management practice it was essential to determine the sensitivity of multiple populations of the stalk borer Chilo partellus (Swinhoe), pink borer Sesamia inferens (Walker) and the cob borer Helicoverpa armigera (Hübner) to Cry1Ab protein through bioassays. The insect populations were collected during growing seasons of Rabi 2005 (October 2005 to February 2006) and Kharif 2006 (May to September 2006). Multiple populations of the three lepidopteran corn pests were found to be susceptible to Cry1Ab. Median lethal concentrations (LC(50)) ranged between 0.008 and 0.068 microg Cry1Ab mL(-1) diet for 18 populations of C. partellus (across two seasons), between 0.12 and 1.99 microg mL(-1) for seven populations of H. armigera and between 0.46 and 0.56 microg mL(-1) for two populations of S. inferens. Dose-response concentrations for lethality and growth inhibition have been determined to mark baseline sensitivity of multiple populations of key lepidopteran corn pests in India to Cry1Ab protein. These benchmark values will be referenced while monitoring resistance to Cry1Ab should Bt corn hybrids expressing Cry1Ab be approved for commercial cultivation in India. Copyright (c) 2010 Society of Chemical Industry.

  13. The VIP Experiment

    International Nuclear Information System (INIS)

    Sirghi, D.L.; Sirghi, F.

    2005-01-01

    The Pauli Exclusion Principle (PEP) is a basic principle of Quantum Mechanics, and its validity has never been seriously challenged. However, given its importance, it is very important to check it as thoroughly as possible. The recently approved VIP (VIolation of Pep) experiment, represents an improved version of the Ramberg and Snow experiment (Ramberg and Snow, Phys. Lett. B238 (1990) 438). VIP shall be performed at the Gran Sasso underground laboratories, and aims to test the Pauli Exclusion Principle for electrons with unprecedented accuracy. VIP is a Collaboration among four Institutions out of three countries (LNF-INFN, and INFN Trieste Italy; SMI-Vienna, Austria; IFIN-HH, Bucharest, Romania). It uses an apparatus with CCDs (Charge Coupled Device) as detectors of X rays - looking for PEP violating transitions in Copper: transitions from the 2p level to 1s with the 1s already occupied by 2 electrons. The characteristic of such transition is the energy - displaced with respect to the normal 2p → 1s one by about 300 eV. VIP will bring the limit on the probability that PEP is violated by electrons to 10 -30 , exploring so a region where new theories allow for a possible PEP violation. (authors)

  14. The New Transgenic cry1Ab/vip3H Rice Poses No Unexpected Ecological Risks to Arthropod Communities in Rice Agroecosystems.

    Science.gov (United States)

    Lu, Zengbin; Dang, Cong; Han, Naishun; Shen, Zhicheng; Peng, Yufa; Stanley, David; Ye, Gongyin

    2016-04-01

    The ecological risks to nontarget organisms should be rigorously assessed before Bt crops are released. Here, the impacts of a new Cry1Ab/Vip3H rice line on arthropod communities in rice agroecosystems were evaluated across 3 yr. Arthropods collected via vacuum were sorted into five guilds. The abundance and proportion of each guild as well as community-level parameters were determined in Cry1Ab/Vip3H and control rice fields. Changes in arthropod species assemblage over sampling dates were investigated by principal response curves (PRCs). Cry1Ab/Vip3H rice did not exert significant impacts on the seasonal density and proportion of each guild, except parasitoids. Detritivore seasonal density, but not its relative abundance, was significantly affected by Cry1Ab/Vip3H rice. Four community indices (species richness S, Shannon-Wiener index H', Simpson index D, and evenness index J') were similar between rice types. PRCs revealed a slight community difference between rice types in the past two tested years, with rice types accounting for 1.0-3.5% of the variance among arthropod communities. However, sampling dates explain 32.1-67.6% for these community differences. Of the 46 taxa with higher species weights, 26.1% of the taxa were significantly different, including seven taxa with higher abundance and five with lower density in Cry1Ab/Vip3H rice fields. These differences may be attributed to change in abundance of prey or hosts but not to direct effects of Bt proteins. We infer that this new Cry1Ab/Vip3H rice line poses no unintended ecological risks to the arthropod community. © 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.

  15. Deletion of aprA and nprA genes for alkaline protease A and neutral protease A from bacillus thuringiensis: effect on insecticidal crystal proteins.

    Science.gov (United States)

    Tan, Y; Donovan, W P

    2001-11-17

    The aprA gene encoding alkaline protease A (AprA) was cloned from Bacillus thuringiensis subsp. kurstaki, and the cloned gene was used to construct aprA-deleted (aprA1) strains of B. thuringiensis. An aprA1 strain of B. thuringiensis that contained the wild-type gene for neutral protease A (nprA(+)) displayed levels of extracellular proteolytic activity that were similar to those of an aprA(+)nprA(+) strain. However, when EDTA was included in the protease assay to inhibit NprA activity the aprA1nprA(+) strain displayed only 2% of the extracellular proteolytic activity of the aprA(+)nprA(+) strain. A strain that was deleted for both aprA and nprA (aprA1nprA3 strain) failed to produce detectable levels of proteolytic activity either in the presence or absence of EDTA in the assay. Compared with the aprA(+)nprA(+) strain the aprA1nprA(+) strain yielded 10% more full-length Cry1Bb crystal protein and the aprA1nprA3 strain yielded 25% more full-length Cry1Bb protein. No significant differences were seen in the 50% lethal dose of Cry1Bb protein from aprA(+)nprA(+) and aprA1nprA3 strains against three species of lepidopteran insects. These results suggest that enhanced yield of certain crystal proteins can be obtained by deletion of the genes aprA and nprA which are the major extracellular proteases of B. thuringiensis.

  16. Efficacy of Insecticide and Bioinsecticide Ground Sprays to Control Metisa plana Walker (Lepidoptera: Psychidae) in Oil Palm Plantations, Malaysia.

    Science.gov (United States)

    Salim, Hasber; Rawi, Che Salmah Md; Ahmad, Abu Hassan; Al-Shami, Salman Abdo

    2015-12-01

    The effectiveness of the synthetic insecticides trichlorfon, lambda-cyhalothrin, cypermethrin emulsion concentrated (EC) and cypermethrin emulsion water based (EW) and a bio-insecticide, Bacillus thuringiensis subsp. kurstaki (Btk), was evaluated at 3, 7, 14 and 30 days after treatment (DAT) for the control of Metisa plana larvae in an oil palm (Elaeis guineensis) plantation in Malaysia. Although all synthetic insecticides effectively reduced the larval population of M. plana, trichlorfon, lambda-cyhalothrin and cypermethrin EC were the fastest-acting. The larval population dropped below the economic threshold level (ETL) 30 days after a single application of the synthetic insecticides. Application of Btk, however, gave poor results, with the larval population remaining above the ETL post treatment. In terms of operational productivity, ground spraying using power spray equipment was time-consuming and resulted in poor coverage. Power spraying may not be appropriate for controlling M. plana infestations in large fields. Using a power sprayer, one man could cover 2-3 ha per day. Hence, power spraying is recommended during outbreaks of infestation in areas smaller than 50 ha.

  17. Effect of ultraviolet and gamma rays on the activity of delta-endotoxin protein crystals of Bacillus thuringiensis

    International Nuclear Information System (INIS)

    Burges, H.D.; Hillyer, S.; Chanter, D.O.

    1975-01-01

    Sensitive bioassays with larvae of Pieris brassicae revealed no reduction of insecticidal activity as a result of severe gamma or ultraviolet irradiation of crystals of Bacillus thuringiensis (serotype V). The measured response was the inhibition of larval feeding by the crystals over exposure periods short enough for the presence of live spores not to influence feeding. The results were analyzed using a logistic model. (U.S.)

  18. Inheritance of Cry1F resistance, cross-resistance and frequency of resistant alleles in Spodoptera frugiperda (Lepidoptera: Noctuidae).

    Science.gov (United States)

    Vélez, A M; Spencer, T A; Alves, A P; Moellenbeck, D; Meagher, R L; Chirakkal, H; Siegfried, B D

    2013-12-01

    Transgenic maize, Zea maize L., expressing the Cry1F protein from Bacillus thuringiensis has been registered for Spodoptera frugiperda (J. E. Smith) control since 2003. Unexpected damage to Cry1F maize was reported in 2006 in Puerto Rico and Cry1F resistance in S. frugiperda was documented. The inheritance of Cry1F resistance was characterized in a S. frugiperda resistant strain originating from Puerto Rico, which displayed >289-fold resistance to purified Cry1F. Concentration-response bioassays of reciprocal crosses of resistant and susceptible parental populations indicated that resistance is recessive and autosomal. Bioassays of the backcross of the F1 generation crossed with the resistant parental strain suggest that a single locus is responsible for resistance. In addition, cross-resistance to Cry1Aa, Cry1Ab, Cry1Ac, Cry1Ba, Cry2Aa and Vip3Aa was assessed in the Cry1F-resistant strain. There was no significant cross-resistance to Cry1Aa, Cry1Ba and Cry2Aa, although only limited effects were observed in the susceptible strain. Vip3Aa was highly effective against susceptible and resistant insects indicating no cross-resistance with Cry1F. In contrast, low levels of cross-resistance were observed for both Cry1Ab and Cry1Ac. Because the resistance is recessive and conferred by a single locus, an F1 screening assay was used to measure the frequency of Cry1F-resistant alleles from populations of Florida and Texas in 2010 and 2011. A total frequency of resistant alleles of 0.13 and 0.02 was found for Florida and Texas populations, respectively, indicating resistant alleles could be found in US populations, although there have been no reports of reduced efficacy of Cry1F-expressing plants.

  19. VIP and VIP gene silencing modulation of differentiation marker N-cadherin and cell shape of corneal endothelium in human corneas ex vivo.

    Science.gov (United States)

    Koh, Shay-Whey M; Chandrasekara, Krish; Abbondandolo, Cara J; Coll, Timothy J; Rutzen, Allan R

    2008-08-01

    Vasoactive intestinal peptide (VIP) is expressed by corneal endothelial (CE) cells and is present in the aqueous humor, which bathes CE cells in vivo. This study demonstrated the role of CE cell VIP in maintaining the expression level of a CE differentiation marker, N-cadherin, and the hexagonal cell shape. To determine the most effective VIP concentration, bovine corneoscleral explants were treated with 0 (control) and 10(-12) to 10(-6) M VIP. Paired human corneas (nine donors) from an eye bank were used as control; the other corneas were treated with VIP. To silence endogenous VIP, paired fresh human donor corneas (from seven cadavers) were transduced with VIP shRNA or the control lentiviral particles and then bisected/quartered for quantitative analysis by semiquantitative RT-PCR (for mRNA) and Western blot analysis/immunocytochemistry (for protein), whereas alizarin red S staining revealed CE cell shape. VIP concentration dependently increased bovine CE cell N-cadherin mRNA levels, with the maximal effect observed between 10(-10) (1.47 +/- 0.06-fold; P = 0.002) and 10(-8) M VIP (1.48 +/- 0.18-fold; P = 0.012). VIP (10(-8) M) treatment increased N-cadherin protein levels in bovine and human CE cells to 1.98 +/- 0.28-fold (P = 0.005) and 1.17 +/- 0.10 (range, 0.91-1.87)-fold (P = 0.050) of their respective controls. VIP antagonist (SN)VIPhyb diminished the VIP effect. VIP silencing resulted in deterioration of the hexagonal cell shape and decreased levels of VIP protein and mRNA, N-cadherin (but not connexin-43) mRNA and protein, and the antiapoptotic Bcl-2 protein. Through its autocrine VIP, CE cells play an active role in maintaining the differentiated state and suppressing apoptosis in the corneal endothelium in situ.

  20. Risk assessment of the biological plant protection product Turex 50 WG, with the organism Bacillus thuringiensis ssp. aizawai CG-91. Opinion of the Panel on Plant Protection Products of the Norwegian Scientific Committee for Food Safety

    OpenAIRE

    Källqvist, Torsten; Dirven, Hubert; Gjøen, Tor; Tronsmo, Arne; Yazdankhah, Siamak Pour; Rivedal, Edgar; Borgå, Katrine; Eklo, Ole Martin; Grung, Merete; Lyche, Jan Ludvig; Låg, Marit; Nilsen, Asbjørn Magne; Sverdrup, Line Emilie

    2016-01-01

    Bacillus thuringiensis are anaerobic, gram-positive bacteria that produce parasporal crystalline protein inclusions, δ-endotoxin, which are toxic to certain invertebrates, especially larvae belonging to the insect orders Coleoptera, Diptera and Lepidoptera. Different strains of Bacillus thuringiensis have therefore a long standing history as plant protective insecticides in many countries, but have not been approved for use in Norway. The Norwegian Scientific Committee for Food Safety (Vitens...

  1. Activity of Bacillus thuringiensis D(delta)-endotoxins against codling moth (Cydia pomonella L.) larvae

    NARCIS (Netherlands)

    Boncheva, R.; Dukiandjiev, S.; Minkov, I.; Maagd, de R.A.; Naimov, S.

    2006-01-01

    Solubilized protoxins of nine Cry1 and one hybrid Cry1 ¿-endotoxin from Bacillus thuringiensis were tested for their activity against larvae of the codling moth (Cydia pomonella L). Cry1Da was the most toxic, followed by Cry1Ab, Cry1Ba, and Cry1Ac, while Cry1Aa, Cry1Fa, Cry1Ia, and SN19 were still

  2. Comparative analysis of Bacillus thuringiensis toxin binding to gypsy moth, browntail moth, and douglas-fir tussock moth midgut tissue sections using fluorescence microscopy

    Science.gov (United States)

    Algimantas P. Valaitis; John D. Podgwaite

    2011-01-01

    Many strains of Bacillus thuringiensis (Bt) produce insecticidal proteins, also referred to as Cry toxins, in crystal inclusions during sporulation. When ingested by insects, the Cry toxins bind to receptors on the brush border midgut epithelial cells and create pores in the epithelial gut membranes resulting in the death of...

  3. SELECTIVITY OF INSECTICIDES TO PREDATORS OF PESTS COTTON PLANT SELETIVIDADE DE INSETICIDAS AOS PREDADORES DAS PRAGAS DO ALGODOEIRO

    Directory of Open Access Journals (Sweden)

    Julio Cezar Silveira Nunes

    2007-09-01

    Full Text Available

    The selectivity of insecticides for the complex of predators of the pests of cotton plant was evaluated in field experiment, in Goiânia- Goiás (Brazil, during the crop 1998/99. The experimental design was the randomized blocks with seven treatments and four repetitions (check, clorfluazuron, Bacillus thuringiensis, alanycarb, endosulfan and acephate in two amounts. The samplings were accomplished in beforeapplication, two days, seven and fourteen days after the treatment. For the obtained results (Henderson & Tilton, the products, in the decreasing order of selectivity, were: alanycarb, clorfluazuron, B. thuringiensis, endosulfan e acephate.

    KEY-WORDS: Insecta; insecticides; cotton plant; predators.

    A seletividade de inseticidas para o complexo das pragas do algodoeiro foi avaliada em experimento de campo, em Goiânia (GO, durante a safra 1998/99. O delineamento experimental foi em blocos ao acaso com sete tratamentos testemunha, clorfluazuron, B. thuringiensis, alanycarb, endosulfan e acephate em duas dosagens, em quatro repetições. As amostragens foram realizadas em pré-aplicação; aos dois, sete e quatorze dias após as pulverizações. Pelos resultados obtidos (fórmula de Herderson & Tilton, os produtos, na ordem decrescente de seletividade, foram: alanycarb, clorfluazuron, B. thuringiensis, endosulfan e acephate.

    PALAVRAS-CHAVE: Insecta; inseticidas; algodão; predadores.

  4. Changes in protease activity and Cry3Aa toxin binding in the Colorado potato beetle: implications for insect resistance to Bacillus thuringiensis toxins

    Science.gov (United States)

    Olga Loseva; Mohamed Ibrahim; Mehmet Candas; C. Noah Koller; Leah S. Bauer; Lee A. Jr. Bulla

    2002-01-01

    Widespread commercial use of Bacillus thuringiensis Cry toxins to control pest insects has increased the likelihood for development of insect resistance to this entomopathogen. In this study, we investigated protease activity profiles and toxin-binding capacities in the midgut of a strain of Colorado potato beetle (CPB) that has developed resistance...

  5. Poly-β-hydroxybutyrate Metabolism Is Unrelated to the Sporulation and Parasporal Crystal Protein Formation in Bacillus thuringiensis.

    Science.gov (United States)

    Wang, Xun; Li, Zhou; Li, Xin; Qian, Hongliang; Cai, Xia; Li, Xinfeng; He, Jin

    2016-01-01

    Poly-3-hydroxybutyrate (PHB) is a natural polymer synthesized by many bacteria as a carbon-energy storage material. It was accumulated maximally prior to the spore formation but was degraded during the process of sporulation in Bacillus thuringiensis. Intriguingly, B. thuringiensis also accumulates large amounts of insecticidal crystal proteins (ICPs) during sporulation, which requires considerable input of carbon and energy sources. How PHB accumulation affects sporulation and ICP formation remains unclear to date. Intuitively, one would imagine that accumulated PHB provides the energy required for ICP formation. Yet our current data indicate that this is not the case. First, growth curves of the deletion mutants of phaC (encoding the PHB synthase) and phaZ (encoding the PHB depolymerase) were found to be similar to the parent strain BMB171; no difference in growth rate could be observed. In addition we further constructed the cry1Ac10 ICP gene overexpression strains of BMB171 (BMB171-cry), as well as its phaC and phaZ deletion mutants ΔphaC-cry and ΔphaZ-cry to compare their spore and ICP production rates. Again, not much change of ICP production was observed among these strains either. In fact, PHB was still degraded in most ΔphaZ-cry cells as observed by transmission electron microscopy. Together these results indicated that there is no direct association between the PHB accumulation and the sporulation and ICP formation in B. thuringiensis. Some other enzymes for PHB degradation or other energy source may be responsible for the sporulation and/or ICP formation in B. thuringiensis.

  6. Distribution of Voltage-Gated Sodium Channel (Nav) Alleles among the Aedes aegypti Populations In Central Java Province and Its Association with Resistance to Pyrethroid Insecticides.

    Science.gov (United States)

    Sayono, Sayono; Hidayati, Anggie Puspa Nur; Fahri, Sukmal; Sumanto, Didik; Dharmana, Edi; Hadisaputro, Suharyo; Asih, Puji Budi Setia; Syafruddin, Din

    2016-01-01

    The emergence of insecticide resistant Aedes aegypti mosquitoes has hampered dengue control efforts. WHO susceptibility tests, using several pyrethroid compounds, were conducted on Ae. aegypti larvae that were collected and raised to adulthood from Semarang, Surakarta, Kudus and Jepara in Java. The AaNaV gene fragment encompassing kdr polymorphic sites from both susceptible and resistant mosquitoes was amplified, and polymorphisms were associated with the resistant phenotype. The insecticide susceptibility tests demonstrated Ae, aegypti resistance to the pyrethroids, with mortality rates ranging from 1.6%-15.2%. Three non-synonymous polymorphisms (S989P, V1016G and F1534C) and one synonymous polymorphism (codon 982) were detected in the AaNaV gene. Eight AaNaV alleles were observed in specimens from Central Java. Allele 3 (SGF) and allele 7 (PGF) represent the most common alleles found and demonstrated strong associations with resistance to pyrethroids (OR = 2.75, CI: 0.97-7.8 and OR = 7.37, CI: 2.4-22.5, respectively). This is the first report of 8 Ae. aegypti AaNaV alleles, and it indicates the development of resistance in Ae. aegypti in response to pyrethroid insecticide-based selective pressure. These findings strongly suggest the need for an appropriate integrated use of insecticides in the region. The 989P, 1016G and 1534C polymorphisms in the AaNaV gene are potentially valuable molecular markers for pyrethroid insecticide resistance monitoring.

  7. VIP secreting tumours in infancy

    International Nuclear Information System (INIS)

    Davies, R.P.; Slavotinek, J.P.; Dorney, S.F.A.

    1990-01-01

    Vasoactive intestinal polypeptide (VIP) secreting neural crest tumours are an uncommon but important treatable cause of intractable childhood diarrhoea. The radiological appearances of two cases are presented with a review of radiological findings in childhood VIP secreting neural crest tumours. Twenty eight cases of childhood VIP secreting neural crest tumours were reviewed. Nineteen (68%) were ganglioneuroblastomas and nine (32%) were ganglioneuromas. The majority of tumours (66%) were in a paravertebral location in the abdomen indicating that a search for such a tumour should be initiated at this site. Eighteen of the twenty eight cases reviewed discussed relevant radiological investigations. Calcification was detected in 50% of abdominal radiographs. Gut dilatation was often a prominent feature. A mass was detected in 5 of 5 cases where ultrasound findings were reported, and seven of seven cases with CT findings reported. Prior to the availability of CT and ultrasound the most useful investigation was IVU which demonstrated evidence of a mass in 5 of 9 cases. The presence of paravertebral calcification and gut dilatation on the plain radiograph of a child with intractable diarrhoea suggests the presence of a VIP secreting neural crest tumour. If an abdominal tumour is not found in the appropriate clinical setting and VIP levels are elevated, a widespread search of the paravertebral region is indicated. (orig.)

  8. Persistence of detectable insecticidal proteins from Bacillus thuringiensis (Cry) and toxicity after adsorption on contrasting soils

    International Nuclear Information System (INIS)

    Hung, T.P.; Truong, L.V.; Binh, N.D.; Frutos, R.; Quiquampoix, H.; Staunton, S.

    2016-01-01

    Insecticidal Cry, or Bt, proteins are produced by the soil-endemic bacterium, Bacillus thuringiensis and some genetically modified crops. Their environmental fate depends on interactions with soil. Little is known about the toxicity of adsorbed proteins and the change in toxicity over time. We incubated Cry1Ac and Cry2A in contrasting soils subjected to different treatments to inhibit microbial activity. The toxin was chemically extracted and immunoassayed. Manduca sexta was the target insect for biotests. Extractable toxin decreased during incubation for up to four weeks. Toxicity of Cry1Ac was maintained in the adsorbed state, but lost after 2 weeks incubation at 25 °C. The decline in extractable protein and toxicity were much slower at 4 °C with no significant effect of soil sterilization. The major driving force for decline may be time-dependent fixation of adsorbed protein, leading to a decrease in the extraction yield in vitro, paralleled by decreasing solubilisation in the larval gut. - Graphical abstract: Biotest, presenting Cry-contaminated feed to Manduca sexta larvae in individual Perspex boxes. Display Omitted - Highlights: • Toxicity of Cry protein is initially conserved after adsorption on soil. • Toxicity and extractability decline with time, more rapidly at 25 °C than 4 °C. • Similar dynamics of Cry1AC and Cry2A on soil with varying texture and organic C. • Sterilization of soil does not change Cry dynamics or temperature effect in soil. • Cry decline is determined by progressive fixation on soil not microbial breakdown. - Toxicity was initially maintained after adsorption on soil and both extractable Cry and toxicity declined rapidly, more slowly at low temperature, due to different fixation dynamics. Toxicity of Cry protein is initially conserved after adsorption on soil.

  9. Acute toxicities and sublethal effects of some conventional insecticides on Trichogramma chilonis (Hymenoptera: Trichogrammatidae).

    Science.gov (United States)

    Wang, De-Sen; He, Yu-Rong; Guo, Xiang-Ling; Luo, Yong-Li

    2012-08-01

    The acute toxicity of 10 conventional insecticides to adult of Trichogramma chilonis Ishii (Hymenoptera: Trichogrammatidae) was bioassayed by membrane method, and then their sublethal effects on the parasitoid were evaluated in the laboratory. Based on sublethal concentration (LC30) values at 8 h after treatment, we determined that adult T. chilonis were the most susceptible to chlorfenapyr, followed by fipronil, spinosad, avermectins, beta-cypermethrin, and cartap, with lethal concentration (LC)30 values of 0.3133, 0.3269, 1.5408, 3.2961, 6.1469, and 9.021 mg/liter, respectively. The field-recommended concentrations of chlorfluazuron, indoxacarb, Bacillus thuringiensis, and tebufenozide caused Cartap and spinosad also reduced longevity (8 and 7.9 d) and fecundity (110.77 and 117.2) of treated adults, but cartap enhanced the female percentage of F1 offspring (61.6%), resulting a statistical higher R0, r(m), and lambda of treated T. chilonis. In contrast, chlorfluazuron and tebufenozide increased longevity (16.4 and 15.4 d) and fecundity (248 and 256.9) of treated adults but slightly decreased the female percentage of F1 offspring (31.4 and 38.1%). Although chlorfenapyr showed no adverse influence on longevity and fecundity, it remarkably reduced the female percentage of F1 offspring (13.5%), leading to a lower R0, r(m), and lambda of treated T. chilonis. Indoxacarb, B. thuringiensis, and beta-cypermethrin had no obvious sublethal effects on the longevity and fecundity of treated adults. Based on these results, we consider B. thuringienesis, chlorfluazuron, indoxacarb, beta-cypermethrin, and tebufenozide safe to T. chilonis, suggesting that these insecticides are compatible with this parasitoid when being used in the field. However, fipronil, chlorfenapyr, spinosad, and avermectins were very harmful to T. chilonis. Timing of application of these insecticides was critical.

  10. Spatial and temporal distribution of airborne Bacillus thuringiensis var. kurstaki during an aerial spray program for gypsy moth eradication.

    OpenAIRE

    Teschke, K; Chow, Y; Bartlett, K; Ross, A; van Netten, C

    2001-01-01

    We measured airborne exposures to the biological insecticide Bacillus thuringiensis var. kurstaki (Btk) during an aerial spray program to eradicate gypsy moths on the west coast of Canada. We aimed to determine whether staying indoors during spraying reduced exposures, to determine the rate of temporal decay of airborne concentrations, and to determine whether drift occurred outside the spray zone. During spraying, the average culturable airborne Btk concentration measured outdoors within the...

  11. Fat ViP MRI: Virtual Phantom Magnetic Resonance Imaging of water-fat systems.

    Science.gov (United States)

    Salvati, Roberto; Hitti, Eric; Bellanger, Jean-Jacques; Saint-Jalmes, Hervé; Gambarota, Giulio

    2016-06-01

    Virtual Phantom Magnetic Resonance Imaging (ViP MRI) is a method to generate reference signals on MR images, using external radiofrequency (RF) signals. The aim of this study was to assess the feasibility of ViP MRI to generate complex-data images of phantoms mimicking water-fat systems. Various numerical phantoms with a given fat fraction, T2* and field map were designed. The k-space of numerical phantoms was converted into RF signals to generate virtual phantoms. MRI experiments were performed at 4.7T using a multi-gradient-echo sequence on virtual and physical phantoms. The data acquisition of virtual and physical phantoms was simultaneous. Decomposition of the water and fat signals was performed using a complex-based water-fat separation algorithm. Overall, a good agreement was observed between the fat fraction, T2* and phase map values of the virtual and numerical phantoms. In particular, fat fractions of 10.5±0.1 (vs 10% of the numerical phantom), 20.3±0.1 (vs 20%) and 30.4±0.1 (vs 30%) were obtained in virtual phantoms. The ViP MRI method allows for generating imaging phantoms that i) mimic water-fat systems and ii) can be analyzed with water-fat separation algorithms based on complex data. Copyright © 2016 Elsevier Inc. All rights reserved.

  12. Vasoactive intestinal polypeptide (VIP) in the pig pancreas

    DEFF Research Database (Denmark)

    Poulsen, Steen Seier

    1984-01-01

    Vasoactive intestinal polypeptide (VIP) in the pig pancreas is localized to nerves, many of which travel along the pancreatic ducts. VIP stimulates pancreatic fluid and bicarbonate secretion like secretin. Electrical vagal stimulation in the pig causes an atropine-resistant profuse secretion...... of bicarbonate-rich pancreatic juice. In an isolated perfused preparation of the pig pancreas with intact vagal nerve supply, electrical vagal stimulation caused an atropine-resistant release of VIP, which accurately parallelled the exocrine secretion of juice and bicarbonate. Perfusion of the pancreas...... with a potent VIP-antiserum inhibited the effect of vagal stimulation on the exocrine secretion. It is concluded, that VIP is responsible for (at least part of) the neurally controlled fluid and bicarbonate secretion from the pig pancreas....

  13. Vacuum maintenance in vacuum insulation panels exemplified with a staggered beam VIP

    Energy Technology Data Exchange (ETDEWEB)

    Kwon, Jae-Sung; Jang, Choong Hyo; Jung, Haeyong; Song, Tae-Ho [Department of Mechanical Engineering, Korea Advanced Institute of Science and Technology, Guseong-dong 373-1, Yuseong-gu, Daejeon (Korea, Republic of)

    2010-05-15

    Thermal insulation performance of a vacuum insulation panel (VIP) is highly dependent on the inner pressure of the VIP. Long-term vacuum maintenance characteristics are investigated in this study for a VIP with an example of polymer staggered beam structure as the core material. Various gas sources deteriorating the vacuum level in the VIP are investigated based on theoretical models and experiments. Gas permeation occurring through heat-sealed flanges and pinholes in the barrier envelope is the largest gas leakage source. The calculated gas permeation rate is in accordance with the experimental result. To reduce these permeations, a three-side sealing envelope and double enveloping are proposed. Outgassing from the core material and inner surface of the envelope is also critical. It is significantly reduced by a baking pre-treatment in vacuum. When the estimated total gas load exceeds the allowable limit within a few years, a getter material may be applied. Double enveloping structure with a getter is promising as it ensures a lifetime of more than 20 years. (author)

  14. Efficacy of insecticides in fruit borer control and residues on sugar apple fruit

    Directory of Open Access Journals (Sweden)

    Alessandro da Silva Oliveira

    Full Text Available ABSTRACT Bahia is the Brazilian state with the largest production of sugar apple fruits (Annona squamosa L., and fruit borer (Cerconota anonella, Sepp. 1830 is a key crop pest. Insecticides are the main strategy for pest control even though there are no pesticides registered for this crop. This study aimed to assess the efficacy of insecticides to control fruit borer and determine the levels of insecticide residues in sugar apple fruits aiming at requesting the extension of authorization to use insecticide products in this crop. The experiment was conducted in an eight-year-old irrigated orchard (2 × 4 m located in Anagé, Bahia, Brazil. The experimental design was a randomized block design with 10 treatments (three insecticides with three doses and a control with water and 5 replications. Each plot was composed of four plants but only the two central ones were assessed. Insecticides and doses (g a.i. 100 L−1 water were Bacillus thuringiensis: 0.8, 1.7, and 2.5; triflumuron: 2.4, 3.6, and 4.8; and imidacloprid: 4.0, 10.0, and 16.0. Nine sprayings were carried out at fortnightly intervals with a costal sprayer with constant pressure, JA-2 nozzle, and with jet directed to the fruits. Ten assessments were performed in order to observe fruit borer presence in 30 previously marked fruits per plot. Imidacloprid, at the highest studied dose, was the only effective treatment. Analyses of imidacloprid residues, at 21 and 30 days after the highest dose application, indicated levels higher than the maximum limit allowed. Insecticides under the conditions tested do not meet the norms for requesting the extension of authorization to use insecticides for citrus in sugar apple fruits.

  15. Effect of selected insecticides on SF9 insect cell line

    International Nuclear Information System (INIS)

    Saleh, M.; Rahmo, A.; Hajjar, J.

    2013-01-01

    The toxic effect of three insecticides: dimethoate (organophosphate insecticide), acetamiprid (neonicotinoid insecticide) and deltamethrin (pyrethroid insecticide) were evaluated in vitro on cultured Sf9 cell line. Cell growth inhibition was measured by the 3- (4,5- dimethylthiazol - 2-yl) - 2,5 - diphenyl tetrazolium bromide (MTT) assay. Regression Analysis was used to estimate the 20% inhibition of cells growth (IC 20). The IC 20 values obtained for deltamethrin, acetamipridand dimethoate were: 46.8, 61.6 and 68.9 μM, respectively. The proportion of phagocytic cells was positively correlated with the applied concentrations of the insecticides. (author)

  16. Labeling of vasoactive intestinal peptide (VIP) and VIP 10-28 fragment with radioiodine by direct method. Comparative study of the kinetics biodistribution and affinity for neuroendocrine tumor cells; Marcacao do peptideo intestinal vasoativo (VIP) e do fragmento VIP10-28 com radioiodo por metodo direto. Estudo comparativo da cinetica de biodistribuicao e da afinidade por celulas de tumor neuroendocrino

    Energy Technology Data Exchange (ETDEWEB)

    Colturato, Maria Tereza

    2005-07-01

    In the progress of the Nuclear Medicine, many protein based radiopharmaceuticals have been developed in the last years using antibodies and, more recently, biologically active natural peptides or similar synthetic peptides. In the search for agents with specificity for the target tissue in tumors detection, it was verified that small sequences of amino acids may interact with selective sites, with homogenous distribution, fast accumulation in tissues and fast blood clearance when compared to the antibodies. Among the peptides used in the diagnosis of tumors, Vasoactive Intestinal Peptide (VIP) has been studied. VIP labeled with iodine-123 is applied in the images of intestinal adenocarcinoma and endocrine tumors. The molecule of VIP contains two tyrosine residues, in the positions 10 and 22 that are, theoretically, equally susceptible to radioiodination for direct method. The objective of this work was to produce VIP labeled with radioiodine (iodine-123), in order to introduce to the brazilian medical class this radiopharmaceutical of interest for the diagnosis and recurrence of tumors that express specific receptors. In an unpublished way, the work studied the labeling and the kinetic distribution of the VIP fragment (VIP 10-28) and verified its potential as radiopharmaceutical applied in the identification of tumors that express VIP receptors. After the choice of the appropriated technique for labeling VIP and VIP 10-28 with radioiodine, using Ceremonial T as oxidant agent and sodium metabisulfite as reducing agent, the quality control procedures were accomplished (electrophoresis and high performance liquid chromatography, HPLC) for radiochemical purity determination as well as the separation of the radiochemical species obtained. Labeling and quality control procedures applied were efficient and accurate. [{sup 131}I]VIP and [{sup 131}l]VIP 10-28 were obtained with high radiochemical purity (> 95%). The purification studies to remove free radioiodine in the

  17. Radiolabeling and in vitro and in vivo characterization of [18F]FB-[R(8,15,21), L17]-VIP as a PET imaging agent for tumor overexpressed VIP receptors.

    Science.gov (United States)

    Cheng, Dengfeng; Yin, Duanzhi; Li, Gucai; Wang, Mingwei; Li, Shiqiang; Zheng, Mingqiang; Cai, Hancheng; Wang, Yongxian

    2006-12-01

    In an effort to develop a peptide-based radiopharmaceutical for the detection of tumors overexpressed vasoactive intestinal peptide receptors with positron emission tomography, we have prepared a novel [R(8,15,21), L17]-VIP peptide for 18F-labeling. This peptide inhibited 125I-VIP binding to rats lung membranes with high affinity [half-maximal inhibitory concentrations (IC50) of 0.12 nm]. Additionally, [R(8,15,21), L17]-VIP showed higher stability than native vasoactive intestinal peptide in vivo of mice. With N-succinimidyl 4-[18F] fluorobenzoate as labeling prosthetic group, [18F]FB-[R(8,15,21), L17]-VIP was obtained in >99% radiochemical purity within 100 min in decay-for-corrected radiochemical yield of 33.6 +/- 3% (n = 5) and a specific radioactivity 255 GBq/micromol at the end of synthesis. Stability of [18F]FB-[R(8,15,21), L17]-VIP in vitro and in vivo were investigated. Biodistribution of this trace was carried out in mice with induced C26 colorectal tumor. Fast clearance of [18F]FB-[R(8,15,21), L17]-VIP from non-target tissues and specific uptakes by tumors realized higher tumor-to-muscle ratio (3.55) and tumor-to-blood ratio (2.37) 60 min postinjection. Clear difference was observed between the blocking and unblocking experiments in biodistribution and whole body radioautography. [18F]FB-[R(8,15,21), L17]-VIP has demonstrated its potential for diagnosing tumors overexpressed vasoactive intestinal peptide receptors both in vitro and in vivo.

  18. NCBI nr-aa BLAST: CBRC-SARA-01-1804 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-SARA-01-1804 ref|ZP_00743402.1| Sensory box/GGDEF family protein [Bacillus thuringiensis serovar israel...ensis ATCC 35646] gb|EAO52327.1| Sensory box/GGDEF family protein [Bacillus thuringiensis serovar israelensis ATCC 35646] ZP_00743402.1 0.20 36% ...

  19. Co-production of parasporal crystal toxins and antimicrobial ...

    African Journals Online (AJOL)

    Co-production of antimicrobial substances and insecticidal compounds by Bacillus thuringiensis BAR 3 was investigated. The cell free supernatant (CFS) of B. thuringiensis showed inhibitory activities against both Gram positive (B. thuringiensis IFO13866 and Staphylococcus aureus ATCC 25923) and Gram negative ...

  20. Labeling of vasoactive intestinal peptide (VIP) and VIP 10-28 fragment with radioiodine by direct method. Comparative study of the kinetics biodistribution and affinity for neuroendocrine tumor cells

    International Nuclear Information System (INIS)

    Colturato, Maria Tereza

    2005-01-01

    In the progress of the Nuclear Medicine, many protein based radiopharmaceuticals have been developed in the last years using antibodies and, more recently, biologically active natural peptides or similar synthetic peptides. In the search for agents with specificity for the target tissue in tumors detection, it was verified that small sequences of amino acids may interact with selective sites, with homogenous distribution, fast accumulation in tissues and fast blood clearance when compared to the antibodies. Among the peptides used in the diagnosis of tumors, Vasoactive Intestinal Peptide (VIP) has been studied. VIP labeled with iodine-123 is applied in the images of intestinal adenocarcinoma and endocrine tumors. The molecule of VIP contains two tyrosine residues, in the positions 10 and 22 that are, theoretically, equally susceptible to radioiodination for direct method. The objective of this work was to produce VIP labeled with radioiodine (iodine-123), in order to introduce to the brazilian medical class this radiopharmaceutical of interest for the diagnosis and recurrence of tumors that express specific receptors. In an unpublished way, the work studied the labeling and the kinetic distribution of the VIP fragment (VIP 10-28) and verified its potential as radiopharmaceutical applied in the identification of tumors that express VIP receptors. After the choice of the appropriated technique for labeling VIP and VIP 10-28 with radioiodine, using Ceremonial T as oxidant agent and sodium metabisulfite as reducing agent, the quality control procedures were accomplished (electrophoresis and high performance liquid chromatography, HPLC) for radiochemical purity determination as well as the separation of the radiochemical species obtained. Labeling and quality control procedures applied were efficient and accurate. [ 131 I]VIP and [ 131 l]VIP 10-28 were obtained with high radiochemical purity (> 95%). The purification studies to remove free radioiodine in the labeling

  1. Fasting and prolonged exercise increase vasoactive intestinal polypeptide (VIP) in plasma

    DEFF Research Database (Denmark)

    Galbo, H; Hilsted, J; Fahrenkrug, J

    1979-01-01

    and maximal exercise. During 59 h of fasting, VIP increased from 3.6 (0.6--6.6) to 10.2 (6.6--13.8) pmol.1(-1) (p less than 0.05). The concentration of glucose in plasma decreased significantly during the prolonged exercise as well as during fasting. The known metabolic actions of VIP and the demonstrated...

  2. 48 CFR 804.1102 - Vendor Information Pages (VIP) Database.

    Science.gov (United States)

    2010-10-01

    ... (VIP) Database. 804.1102 Section 804.1102 Federal Acquisition Regulations System DEPARTMENT OF VETERANS AFFAIRS GENERAL ADMINISTRATIVE MATTERS Contract Execution 804.1102 Vendor Information Pages (VIP) Database. Prior to January 1, 2012, all VOSBs and SDVOSBs must be listed in the VIP database, available at http...

  3. VIP in human neonates and infants as measured by radioimmunoassay and radioreceptorassay

    International Nuclear Information System (INIS)

    Dupont, C.; Besson, J.; Laburthe, M.; Bataille, D.; Rosselin, G.

    1977-01-01

    The Vasoactive Intestinal Peptide (VIP) was assayed in the gut of human neonates and premature infants immediately after death or surgery. In these conditions, values ranged between 150 and 740ng/g of boiled tissue. The implication of VIP in Vermer-Morrison syndrome is further assessed by the correlation between clinical symptomatology and plasma VIP levels. The immunoassayable VIP (IA-VIP) extracted from normal gut or tumor is shown to fully interact with specific receptor for VIP in liver. This fact suggests the biological potency of IA-VIP [fr

  4. Celebrity Patients, VIPs, and Potentates.

    Science.gov (United States)

    Groves, James E.; Dunderdale, Barbara A.; Stern, Theodore A.

    2002-12-01

    BACKGROUND: During the second half of the 20th century, the literature on the doctor-patient relationship mainly dealt with the management of "difficult" (personality-disordered) patients. Similar problems, however, surround other types of "special" patients. METHOD: An overview and analysis of the literature were conducted. As a result, such patients can be subcategorized by their main presentations; each requires a specific management strategy. RESULTS: Three types of "special" patients stir up irrational feelings in their caregivers. Sick celebrities threaten to focus public scrutiny on the private world of medical caregivers. VIPs generate awe in caregivers, with loss of the objectivity essential to the practice of scientific medicine. Potentates unearth narcissism in the caregiver-patient relationship, which triggers a struggle between power and shame. Pride, privacy, and the staff's need to be in control are all threatened by introduction of the special patient into medicine's closed culture. CONCLUSION: The privacy that is owed to sick celebrities should be extended to protect overexposed staff. The awe and loss of medical objectivity that VIPs generate are counteracted by team leadership dedicated to avoiding any deviation from standard clinical procedure. Moreover, the collective ill will surrounding potentates can be neutralized by reassuring them that they are "special"-and by caregivers mending their own vulnerable self-esteem.

  5. Is VIP1 important for Agrobacterium-mediated transformation?

    Science.gov (United States)

    Shi, Yong; Lee, Lan-Ying; Gelvin, Stanton B

    2014-09-01

    Agrobacterium genetically transforms plants by transferring and integrating T-(transferred) DNA into the host genome. This process requires both Agrobacterium and host proteins. VirE2 interacting protein 1 (VIP1), an Arabidopsis bZIP protein, has been suggested to mediate transformation through interaction with and targeting of VirE2 to nuclei. We examined the susceptibility of Arabidopsis vip1 mutant and VIP1 overexpressing plants to transformation by numerous Agrobacterium strains. In no instance could we detect altered transformation susceptibility. We also used confocal microscopy to examine the subcellular localization of Venus-tagged VirE2 or Venus-tagged VIP1, in the presence or absence of the other untagged protein, in different plant cell systems. We found that VIP1-Venus localized in both the cytoplasm and the nucleus of Arabidopsis roots, agroinfiltrated Nicotiana benthamiana leaves, Arabidopsis mesophyll protoplasts and tobacco BY-2 protoplasts, regardless of whether VirE2 was co-expressed. VirE2 localized exclusively to the cytoplasm of tobacco and Arabidopsis protoplasts, whether in the absence or presence of VIP1 overexpression. In transgenic Arabidopsis plants and agroinfiltrated N. benthamina leaves we could occasionally detect small aggregates of the Venus signal in nuclei, but these were likely to be imagining artifacts. The vast majority of VirE2 remained in the cytoplasm. We conclude that VIP1 is not important for Agrobacterium-mediated transformation or VirE2 subcellular localization. © 2014 The Authors The Plant Journal © 2014 John Wiley & Sons Ltd.

  6. Ecology and diversity of Bacillus thuringiensis in soil environment ...

    African Journals Online (AJOL)

    Bacillus thuringiensis populations ranged between 4.23 x 105, 6.52 x 105 cfu/g soil and consist of 11 types of isolates with 3 polymorphic, 7 spherical and 1 bipyramidal type of crystals. Polymorphic crystal containing isolates were further characterized. B. thuringiensis isolates were circular, white, flat and undulate or entire.

  7. Behavioural actions of vasoactive intestinal peptide (VIP)

    NARCIS (Netherlands)

    Kloet, E.R.; Cottrell, G.A.; Veldhuis, H.D.; Rostene, W.H.

    1984-01-01

    The effect of vasoactive intestinal peptide (VIP) was studied on fear-motivated behaviours, exploration of a novel environment and on novelty and ACTH-induced grooming. VIP was administered via a plastic cannula into the lateral ventricle. Retention of a step-through passive avoidance task was

  8. Biochemical, immunological and toxicological characteristics of the crystal proteins of Bacillus thuringiensis subsp. medellin

    Directory of Open Access Journals (Sweden)

    Sergio Orduz

    1996-04-01

    Full Text Available Characterization of the insecticidal and hemolytic activity of solubilized crystal proteins of Bacillus thuringiensis (Bt subsp. medellin (Btmed was performed and compared to solubilized crystal proteins of isolates 1884 of B. thuringiensis subsp. israelensis (Bti and isolate PG-14 of B. thuringiensis subsp. morrisoni (Btm. In general, at acid pH values solubilization of the Bt crystalline parasporal inclusions (CPI was lower than at alkaline pH. The larvicidal activity demonstrated by the CPI of Btmed indicated that optimal solubilization of CPI takes place at a pH value of 11.3, in Bti at pH values from 5.03 to 11.3 and in Btm at pH values from 9.05 to 11.3. Hemolytic activity against sheep red blood cells was mainly found following extraction at pH 11.3 in all Bt strains tested. Polyacrylamide gel electrophoresis under denaturing conditions revealed that optimal solubilization of the CPI in all Bt strains takes place at the alkaline pH values from 9.05 to 11.3. An enriched preparation of Btmed crystals was obtained, solubilized and crystal proteins were separated on a size exclusion column (Sephacryl S-200. Three main protein peaks were observed on the chromatogram. The first peak had two main proteins that migrate between 90 to 100 kDa. These proteins are apparently not common to other Bt strains isolated to date. The second and third peaks obtained from the size exclusion column yielded polypeptides of 68 and 28-30 kDa, respectively. Each peak independently, showed toxicity against 1st instar Culex quinquefasciatus larvae. Interestingly, combinations of the fractions corresponding to the 68 and 30 kDa protein showed an increased toxicity. These results suggest that the 94 kDa protein is an important component of the Btmed toxins with the highest potency to kill mosquito larvae. When crystal proteins of Bti were probed with antisera raised independently against the three main protein fractions of Btmed, the only crystal protein that showed

  9. Impact of UV radiation on activity of linear furanocoumarins and Bacillus thuringiensis var. kurstaki against Spodoptera exigua: Implications for tritrophic interactions

    Energy Technology Data Exchange (ETDEWEB)

    Trumble, J.T.; Moar, W.J.; Brewer, M.J.; Carson, W.G. (Univ. of California, Riverside (United States))

    1991-05-01

    Acidic fogs with a pH of 2.0 and duration of 2 hr did not reduce the efficacy of Bacillus thuringiensis var. kurstaki. Therefore, the impact of UV radiation was investigated on the interactions between (1) levels of the antibacterial linear furanocoumarins psoralen, bergapten, and xanthotoxin in Apium graveolens (L.) occurring following a 2.0 pH acidic fog episode, (2) the noctuid Spodoptera exigua, and (3) a sublethal dosage of the microbial pathogen B. thuringiensis var. kurstaki. Mean time to pupation in the absence of UV radiation was significantly extended by the addition of either psoralens or B. thuringiensis. Larvae developing on diets containing B. thuringiensis plus psoralens required nearly 40% longer to pupate than controls, but their effects were additive as the interaction was not significant. Mean time to mortality, a weighted average time of death, was not significantly affected by any of the treatments. In a 2 {times} 2 {times} 2 factorial analysis, all main effects reduced survival significantly, as did the three-way interaction. Thus, antagonistic interactions with psoralens that would reduce the effectiveness of B. thuringiensis in the field were not observed. When pairs of main effects were nested within the two levels of the third factor, several two-way interactions were found. Interestingly, the activity of B. thuringiensis and the psoralens, individually or in combination, was enhanced by exposure to UV radiation. Implications of this research are discussed for both natural and agricultural ecosystems.

  10. Impact of UV radiation on activity of linear furanocoumarins and Bacillus thuringiensis var. kurstaki against Spodoptera exigua: Implications for tritrophic interactions

    International Nuclear Information System (INIS)

    Trumble, J.T.; Moar, W.J.; Brewer, M.J.; Carson, W.G.

    1991-01-01

    Acidic fogs with a pH of 2.0 and duration of 2 hr did not reduce the efficacy of Bacillus thuringiensis var. kurstaki. Therefore, the impact of UV radiation was investigated on the interactions between (1) levels of the antibacterial linear furanocoumarins psoralen, bergapten, and xanthotoxin in Apium graveolens (L.) occurring following a 2.0 pH acidic fog episode, (2) the noctuid Spodoptera exigua, and (3) a sublethal dosage of the microbial pathogen B. thuringiensis var. kurstaki. Mean time to pupation in the absence of UV radiation was significantly extended by the addition of either psoralens or B. thuringiensis. Larvae developing on diets containing B. thuringiensis plus psoralens required nearly 40% longer to pupate than controls, but their effects were additive as the interaction was not significant. Mean time to mortality, a weighted average time of death, was not significantly affected by any of the treatments. In a 2 x 2 x 2 factorial analysis, all main effects reduced survival significantly, as did the three-way interaction. Thus, antagonistic interactions with psoralens that would reduce the effectiveness of B. thuringiensis in the field were not observed. When pairs of main effects were nested within the two levels of the third factor, several two-way interactions were found. Interestingly, the activity of B. thuringiensis and the psoralens, individually or in combination, was enhanced by exposure to UV radiation. Implications of this research are discussed for both natural and agricultural ecosystems

  11. Seleção e caracterização de estirpes de Bacillus thuringiensis eficientes contra a Diatraea saccharalis (Lepidoptera: Crambidae

    Directory of Open Access Journals (Sweden)

    Cristina Lima de Macedo

    2012-12-01

    Full Text Available O objetivo deste trabalho foi selecionar e caracterizar estirpes nativas de Bacillus thuringiensis tóxicas a Diatraea saccharalis (Lepidoptera: Crambidae. Cento e seis estirpes pertencentes ao Banco de Bactérias de Invertebrados, da Embrapa Recursos Genéticos e Biotecnologia, foram testadas quanto à toxicidade a D. saccharalis, e, as mais tóxicas, caracterizadas por métodos bioquímicos e moleculares. Das 106 estirpes testadas, 16 causaram 100% de mortalidade em 24 horas. As três estirpes mais tóxicas apresentaram concentração letal média entre 8 e 43 ng cm-2. O perfil proteico das 16 estirpes mostrou a presença de proteínas de 130 e 65 kDa, e a caracterização molecular mostrou a presença dos genes tipo cry1 e cry2: cry1Aa, cry1Ab, cry1Ac e cry2Aa. A concentração letal média de esporos e cristais obtidos a partir de estirpes recombinantes, que expressavam individualmente os genes cry1Aa, cry1Ab, cry1Ac e cry2Aa, variou entre 222 e 610 ng cm-2, valores muito superiores aos das estirpes nativas mais tóxicas, que apresentavam possibilidade de expressão simultânea desses genes. Este resultado é indicativo de que há sinergia entre as toxinas. Há interação entre as toxinas de B. thuringiensis e seus receptores na broca-do-colmo da cana-de-açúcar.

  12. Parasympathetic denervation increases responses to VIP in isolated rat parotid acini

    International Nuclear Information System (INIS)

    McMillian, M.K.; Talamo, B.R.

    1989-01-01

    Vasoactive intestinal peptide (VIP) is a putative neurotransmitter found in the salivary glands of many species, including the rat parotid gland. Parasympathetic denervation has been reported to deplete VIP in the rat parotid gland and to lead to supersensitivity to this peptide in vivo. We have compared the effects of VIP on acini isolated from parasympathetically denervated and unoperated parotid glands to examine possible supersensitivity to the peptide in vitro. VIP normally produced responses similar to those obtained with a low concentration of the beta adrenergic agonist isoproterenol (ISO), but strikingly different from the effects obtained with the muscarinic agonist carbachol (CARB). In parotid membrane preparations, VIP stimulated adenylate cyclase activity. Dissociated acini treated with VIP showed increases in cAMP accumulation and amylase release which were potentiated by forskolin and also by inhibition of phosphodiesterase. After parasympathetic denervation, maximal effects of VIP on adenylate cyclase, cAMP accumulation and amylase release in intact cells were increased two- to five-fold over contralateral control (or unoperated) parotid responses. The increase in adenylate cyclase-mediated responses after denervation was specific to VIP; there was no increased response nor increased sensitivity of any of these responses to ISO. Specific [125I]VIP binding to parotid acini increased two-fold per gland and three-fold per mg of protein after denervation; this probably explains the observed increases in the response to VIP

  13. Encoding of movement in near extrapersonal space in primate area VIP

    Directory of Open Access Journals (Sweden)

    Frank eBremmer

    2013-02-01

    Full Text Available Many neurons in the macaque ventral intraparietal area (VIP are multimodal, i.e., they respond not only to visual but also to tactile, auditory and vestibular stimulation. Anatomical studies have shown distinct projections between area VIP and a region of premotor cortex controlling head movements. A specific function of area VIP could be to guide movements in order to head for and/or to avoid objects in near extra-personal space. This behavioral role would require a consistent representation of visual motion within 3-D space and enhanced activity for nearby motion signals. Accordingly, in our present study we investigated whether neurons in area VIP are sensitive to moving visual stimuli containing depth signals from horizontal disparity. We recorded single unit activity from area VIP of two awake behaving monkeys (M. mulatta fixating a central target on a projection screen. Sensitivity of neurons to horizontal disparity was assessed by presenting large field moving images (random dot fields stereoscopically to the two eyes by means of LCD shutter goggles synchronized with the stimulus computer. During an individual trial, stimuli had one of seven different disparity values ranging from 3 degrees uncrossed- (far to 3 degrees crossed- (near disparity in 1 degree steps. Stimuli moved at constant speed in all simulated depth planes. Different disparity values were presented across trials in pseudo-randomized order. 61% percent of the motion sensitive cells had a statistically significant selectivity for the horizontal disparity of the stimulus (p<0.05, distribution free ANOVA. 75% of them preferred crossed-disparity values, i.e. moving stimuli in near space, with the highest mean activity for the nearest stimulus. At the population level, preferred direction of visual stimulus motion was not affected by horizontal disparity. Thus, our findings are in agreement with the behavioral role of area VIP in the representation of movement in near extra

  14. Sodium Solute Symporter and Cadherin Proteins Act as Bacillus thuringiensis Cry3Ba Toxin Functional Receptors in Tribolium castaneum*

    Science.gov (United States)

    Contreras, Estefanía; Schoppmeier, Michael; Real, M. Dolores; Rausell, Carolina

    2013-01-01

    Understanding how Bacillus thuringiensis (Bt) toxins interact with proteins in the midgut of susceptible coleopteran insects is crucial to fully explain the molecular bases of Bt specificity and insecticidal action. In this work, aminopeptidase N (TcAPN-I), E-cadherin (TcCad1), and sodium solute symporter (TcSSS) have been identified by ligand blot as putative Cry3Ba toxin-binding proteins in Tribolium castaneum (Tc) larvae. RNA interference knockdown of TcCad1 or TcSSS proteins resulted in decreased susceptibility to Cry3Ba toxin, demonstrating the Cry toxin receptor functionality for these proteins. In contrast, TcAPN-I silencing had no effect on Cry3Ba larval toxicity, suggesting that this protein is not relevant in the Cry3Ba toxin mode of action in Tc. Remarkable features of TcSSS protein were the presence of cadherin repeats in its amino acid sequence and that a TcSSS peptide fragment containing a sequence homologous to a binding epitope found in Manduca sexta and Tenebrio molitor Bt cadherin functional receptors enhanced Cry3Ba toxicity. This is the first time that the involvement of a sodium solute symporter protein as a Bt functional receptor has been demonstrated. The role of this novel receptor in Bt toxicity against coleopteran insects together with the lack of receptor functionality of aminopeptidase N proteins might account for some of the differences in toxin specificity between Lepidoptera and Coleoptera insect orders. PMID:23645668

  15. Insecticide treated curtains and residual insecticide treatment to control Aedes aegypti: An acceptability study in Santiago de Cuba.

    Directory of Open Access Journals (Sweden)

    Dennis Pérez

    2018-01-01

    Full Text Available Within the context of a field trial conducted by the Cuban vector control program (AaCP, we assessed acceptability of insecticide-treated curtains (ITCs and residual insecticide treatment (RIT with deltamethrin by the community. We also assessed the potential influence of interviewees' risk perceptions for getting dengue and disease severity.We embedded a qualitative study using in-depth interviews in a cluster randomized trial (CRT testing the effectiveness of ITCs and RIT in Santiago de Cuba. In-depth interviews (N = 38 were conducted four and twelve months after deployment of the tools with people who accepted the tools, who stopped using them and who did not accept the tools. Data analysis was deductive. Main reasons for accepting ITCs at the start of the trial were perceived efficacy and not being harmful to health. Constraints linked to manufacturer instructions were the main reason for not using ITCs. People stopped using the ITCs due to perceived allergy, toxicity and low efficacy. Few heads of households refused RIT despite the noting reasons for rejection, such as allergy, health hazard and toxicity. Positive opinions of the vector control program influenced acceptability of both tools. However, frequent insecticide fogging as part of routine AaCP vector control actions diminished perceived efficacy of both tools and, therefore, acceptability. Fifty percent of interviewees did feel at risk for getting dengue and considered dengue a severe disease. However, this did not appear to influence acceptability of ITCs or RIT.Acceptability of ITCs and RIT was linked to acceptability of AaCP routine vector control activities. However, uptake and use were not always an indication of acceptability. Factors leading to acceptability may be best identified using qualitative methods, but more research is needed on the concept of acceptability and its measurement.

  16. Insecticide treated curtains and residual insecticide treatment to control Aedes aegypti: An acceptability study in Santiago de Cuba.

    Science.gov (United States)

    Pérez, Dennis; Van der Stuyft, Patrick; Toledo, María Eugenia; Ceballos, Enrique; Fabré, Francisco; Lefèvre, Pierre

    2018-01-01

    Within the context of a field trial conducted by the Cuban vector control program (AaCP), we assessed acceptability of insecticide-treated curtains (ITCs) and residual insecticide treatment (RIT) with deltamethrin by the community. We also assessed the potential influence of interviewees' risk perceptions for getting dengue and disease severity. We embedded a qualitative study using in-depth interviews in a cluster randomized trial (CRT) testing the effectiveness of ITCs and RIT in Santiago de Cuba. In-depth interviews (N = 38) were conducted four and twelve months after deployment of the tools with people who accepted the tools, who stopped using them and who did not accept the tools. Data analysis was deductive. Main reasons for accepting ITCs at the start of the trial were perceived efficacy and not being harmful to health. Constraints linked to manufacturer instructions were the main reason for not using ITCs. People stopped using the ITCs due to perceived allergy, toxicity and low efficacy. Few heads of households refused RIT despite the noting reasons for rejection, such as allergy, health hazard and toxicity. Positive opinions of the vector control program influenced acceptability of both tools. However, frequent insecticide fogging as part of routine AaCP vector control actions diminished perceived efficacy of both tools and, therefore, acceptability. Fifty percent of interviewees did feel at risk for getting dengue and considered dengue a severe disease. However, this did not appear to influence acceptability of ITCs or RIT. Acceptability of ITCs and RIT was linked to acceptability of AaCP routine vector control activities. However, uptake and use were not always an indication of acceptability. Factors leading to acceptability may be best identified using qualitative methods, but more research is needed on the concept of acceptability and its measurement.

  17. Bacillus thuringiensis: fermentation process and risk assessment: a short review

    Directory of Open Access Journals (Sweden)

    Deise M. F Capalbo

    1995-02-01

    Full Text Available Several factors make the local production of Bacillus thuringiensis (Bt highly appropriate for pest control in developing nations. Bt can be cheaply produced on a wide variety of low cost, organic substrates. Local production results in considerable savings in hard currency which otherwise would be spent on importation of chemical and biological insecticides. The use of Bt in Brazil has been limited in comparison with chemical insecticides. Although Bt is imported, some Brazilian researchers have been working on its development and production. Fermentation processes (submerged and semi-solid were applied, using by-products from agro-industries. As the semi-solid fermentation process demonstrated to be interesting for Bt endotoxins production, it could be adopted for small scale local production. Although promising results had been achieved, national products have not been registered due to the absence of a specific legislation for biological products. Effective actions are being developed in order to solve this gap. Regardless of the biocontrol agents being considered atoxic and harmless to the environment, information related to direct and indirect effects of microbials are still insufficient in many cases. The risk analysis of the use of microbial control agents is of upmost importance nowadays, and is also discussed.

  18. VIP: Vortex Image Processing Package for High-contrast Direct Imaging

    Science.gov (United States)

    Gomez Gonzalez, Carlos Alberto; Wertz, Olivier; Absil, Olivier; Christiaens, Valentin; Defrère, Denis; Mawet, Dimitri; Milli, Julien; Absil, Pierre-Antoine; Van Droogenbroeck, Marc; Cantalloube, Faustine; Hinz, Philip M.; Skemer, Andrew J.; Karlsson, Mikael; Surdej, Jean

    2017-07-01

    We present the Vortex Image Processing (VIP) library, a python package dedicated to astronomical high-contrast imaging. Our package relies on the extensive python stack of scientific libraries and aims to provide a flexible framework for high-contrast data and image processing. In this paper, we describe the capabilities of VIP related to processing image sequences acquired using the angular differential imaging (ADI) observing technique. VIP implements functionalities for building high-contrast data processing pipelines, encompassing pre- and post-processing algorithms, potential source position and flux estimation, and sensitivity curve generation. Among the reference point-spread function subtraction techniques for ADI post-processing, VIP includes several flavors of principal component analysis (PCA) based algorithms, such as annular PCA and incremental PCA algorithms capable of processing big datacubes (of several gigabytes) on a computer with limited memory. Also, we present a novel ADI algorithm based on non-negative matrix factorization, which comes from the same family of low-rank matrix approximations as PCA and provides fairly similar results. We showcase the ADI capabilities of the VIP library using a deep sequence on HR 8799 taken with the LBTI/LMIRCam and its recently commissioned L-band vortex coronagraph. Using VIP, we investigated the presence of additional companions around HR 8799 and did not find any significant additional point source beyond the four known planets. VIP is available at http://github.com/vortex-exoplanet/VIP and is accompanied with Jupyter notebook tutorials illustrating the main functionalities of the library.

  19. Proteome response of Tribolium castaneum larvae to Bacillus thuringiensis toxin producing strains.

    Directory of Open Access Journals (Sweden)

    Estefanía Contreras

    Full Text Available Susceptibility of Tribolium castaneum (Tc larvae was determined against spore-crystal mixtures of five coleopteran specific and one lepidopteran specific Bacillus thuringiensis Cry toxin producing strains and those containing the structurally unrelated Cry3Ba and Cry23Aa/Cry37Aa proteins were found toxic (LC(50 values 13.53 and 6.30 µg spore-crystal mixture/µL flour disc, respectively. Using iTRAQ combined with LC-MS/MS allowed the discovery of seven novel differentially expressed proteins in early response of Tc larvae to the two active spore-crystal mixtures. Proteins showing a statistically significant change in treated larvae compared to non-intoxicated larvae fell into two major categories; up-regulated proteins were involved in host defense (odorant binding protein C12, apolipophorin-III and chemosensory protein 18 and down-regulated proteins were linked to metabolic pathways affecting larval metabolism and development (pyruvate dehydrogenase Eα subunit, cuticular protein, ribosomal protein L13a and apolipoprotein LI-II. Among increased proteins, Odorant binding protein C12 showed the highest change, 4-fold increase in both toxin treatments. The protein displayed amino acid sequence and structural homology to Tenebrio molitor 12 kDa hemolymph protein b precursor, a non-olfactory odorant binding protein. Analysis of mRNA expression and mortality assays in Odorant binding protein C12 silenced larvae were consistent with a general immune defense function of non-olfactory odorant binding proteins. Regarding down-regulated proteins, at the transcriptional level, pyruvate dehydrogenase and cuticular genes were decreased in Tc larvae exposed to the Cry3Ba producing strain compared to the Cry23Aa/Cry37Aa producing strain, which may contribute to the developmental arrest that we observed with larvae fed the Cry3Ba producing strain. Results demonstrated a distinct host transcriptional regulation depending upon the Cry toxin treatment. Knowledge

  20. Exploration of Bacillus thuringiensis Berl. from soil and screening test its toxicity on insects of Lepidoptera order

    Science.gov (United States)

    Astuti, DT; Pujiastuti, Y.; Suparman, SHK; Damiri, N.; Nugraha, S.; Sembiring, ER; Mulawarman

    2018-01-01

    Bacillus thuringiensis is a gram-positive bacterium that produces crystal proteins toxic (ᴕ-endotoxin) specific to the target insect, but is not toxic to humans and non-target organisms. This study aims to explore the origin of the soil bacterium B. thuringiensis sub-district Sekayu, Banyuasin, South Sumatra and toxicity to larvae of lepidoptera. Fifty soil samples were taken from Musi Banyuasin District, namely 15 from Kayuare strip 2, 20 from Kayuare and 15 from Lumpatan. Isolation, characterization, identification and screening test were conducted in the laboratorium of Pest and Disease, Agricultural Faculty, Sriwijaya University. Isolat codes were given based on the area origin of the samples. Results of the study showed that from 50 isolates of bacteria that had been isolated, there were 15 bacterial isolates, characterized by morphology and physiology the same as B. thuringiensis, which has round colonies, white, wrinkled edges, slippery, elevation arise, aerobic and gram-positive. Of the 15 codes that contain positive isolates of B. thuringiensis, we have obtained several isolates of the following codes: KJ2D5, KJ2N1, KJ2N4, KJ2B3, KJ3R1, KJ3R2, KJ3R3, KJ3R5, KJ3J3, KJ3J4, KJ3P1, DLM5, DLKK12, and DLKK23. Results of screening tests on insects of the Lepidoptera Order showed that there were six isolates that had toxic to Plutella xylostella and Spodoptera litura insects, ie bacterial isolate codes DLM5, KJ3R3, KJ3R5, KJ3J4, KJ3P1, and DLKK23.

  1. VIP/PACAP receptor mediation of cutaneous active vasodilation during heat stress in humans.

    Science.gov (United States)

    Kellogg, Dean L; Zhao, Joan L; Wu, Yubo; Johnson, John M

    2010-07-01

    Vasoactive intestinal peptide (VIP) is implicated in cutaneous active vasodilation in humans. VIP and the closely related pituitary adenylate cyclase activating peptide (PACAP) act through several receptor types: VIP through VPAC1 and VPAC2 receptors and PACAP through VPAC1, VPAC2, and PAC1 receptors. We examined participation of VPAC2 and/or PAC1 receptors in cutaneous vasodilation during heat stress by testing the effects of their specific blockade with PACAP6-38. PACAP6-38 dissolved in Ringer's was administered by intradermal microdialysis at one forearm site while a control site received Ringer's solution. Skin blood flow was monitored by laser-Doppler flowmetry (LDF). Blood pressure was monitored noninvasively and cutaneous vascular conductance (CVC) calculated. A 5- to 10-min baseline period was followed by approximately 70 min of PACAP6-38 (100 microM) perfusion at one site in normothermia and a 3-min period of body cooling. Whole body heating was then performed to engage cutaneous active vasodilation and was maintained until CVC had plateaued at an elevated level at all sites for 5-10 min. Finally, 58 mM sodium nitroprusside was perfused through both microdialysis sites to effect maximal vasodilation. No CVC differences were found between control and PACAP6-38-treated sites during normothermia (19 +/- 3%max untreated vs. 20 +/- 3%max, PACAP6-38 treated; P > 0.05 between sites) or cold stress (11 +/- 2%max untreated vs. 10 +/- 2%max, PACAP6-38 treated, P > 0.05 between sites). PACAP6-38 attenuated the increase in CVC during whole body heating when compared with untreated sites (59 +/- 3%max untreated vs. 46 +/- 3%max, PACAP6-38 treated, P < 0.05). We conclude that VPAC2 and/or PAC1 receptor activation is involved in cutaneous active vasodilation in humans.

  2. VIP and PACAP

    DEFF Research Database (Denmark)

    Fahrenkrug, Jan

    2010-01-01

    amounts. Carboxyamidation of VIP and PHI is not critical and glycine-extended forms of both peptides have been demonstrated. Pituitary adenylate cyclase activating polypeptide (PACAP) is derived from a 170 amino acid long precursor, which gives rise to PACAP 38, PACAP 27 and PACAP related peptide (PRP...

  3. Enhancing emotion recognition in VIPs with haptic feedback

    NARCIS (Netherlands)

    Buimer, Hendrik; Bittner, Marian; Kostelijk, Tjerk; van der Geest, Thea; van Wezel, Richard Jack Anton; Zhao, Yan; Stephanidis, Constantine

    2016-01-01

    The rise of smart technologies has created new opportunities to support blind and visually impaired persons (VIPs). One of the biggest problems we identified in our previous research on problems VIPs face during activities of daily life concerned the recognition of persons and their facial

  4. Effect of supplementation of arachidonic acid (AA) or a combination of AA plus docosahexaenoic acid on breastmilk fatty acid composition

    NARCIS (Netherlands)

    Smit, EN; Koopmann, M; Boersma, ER; Muskiet, FAJ

    We investigated whether supplementation with arachidonic acid (20:4 omega 6; AA), ora combination of AA and docosahexaenoic acid (22:6 omega 3; DHA) would affect human milk polyunsaturated fatty acid (PUFA) composition. Ten women were daily supplemented with 300 mg AA, eight with 300 mg AA, 110 mg

  5. NCBI nr-aa BLAST: CBRC-CBRE-01-1192 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available cillus thuringiensis serovar israelensis ATCC 35646] gb|EAO55692.1| 3-oxoacyl-[acyl-carrier protein] reducta...se [Bacillus thuringiensis serovar israelensis ATCC 35646] ZP_00740054.1 3.1 38% ...

  6. A "genome-to-lead" approach for insecticide discovery: pharmacological characterization and screening of Aedes aegypti D(1-like dopamine receptors.

    Directory of Open Access Journals (Sweden)

    Jason M Meyer

    2012-01-01

    Full Text Available BACKGROUND: Many neglected tropical infectious diseases affecting humans are transmitted by arthropods such as mosquitoes and ticks. New mode-of-action chemistries are urgently sought to enhance vector management practices in countries where arthropod-borne diseases are endemic, especially where vector populations have acquired widespread resistance to insecticides. METHODOLOGY/PRINCIPAL FINDINGS: We describe a "genome-to-lead" approach for insecticide discovery that incorporates the first reported chemical screen of a G protein-coupled receptor (GPCR mined from a mosquito genome. A combination of molecular and pharmacological studies was used to functionally characterize two dopamine receptors (AaDOP1 and AaDOP2 from the yellow fever mosquito, Aedes aegypti. Sequence analyses indicated that these receptors are orthologous to arthropod D(1-like (Gα(s-coupled receptors, but share less than 55% amino acid identity in conserved domains with mammalian dopamine receptors. Heterologous expression of AaDOP1 and AaDOP2 in HEK293 cells revealed dose-dependent responses to dopamine (EC(50: AaDOP1 = 3.1±1.1 nM; AaDOP2 = 240±16 nM. Interestingly, only AaDOP1 exhibited sensitivity to epinephrine (EC(50 = 5.8±1.5 nM and norepinephrine (EC(50 = 760±180 nM, while neither receptor was activated by other biogenic amines tested. Differential responses were observed between these receptors regarding their sensitivity to dopamine agonists and antagonists, level of maximal stimulation, and constitutive activity. Subsequently, a chemical library screen was implemented to discover lead chemistries active at AaDOP2. Fifty-one compounds were identified as "hits," and follow-up validation assays confirmed the antagonistic effect of selected compounds at AaDOP2. In vitro comparison studies between AaDOP2 and the human D(1 dopamine receptor (hD(1 revealed markedly different pharmacological profiles and identified amitriptyline and doxepin as AaDOP2

  7. VIP : A Visual Editor and Compiler for v-Promela

    OpenAIRE

    Kamel, Moataz; Leue, Stefan

    2001-01-01

    We describe the Visual Interface to Promela (VIP) tool that we have recently implemented. VIP supports the visual editing and maintenance of v-Promela models. v-Promela is a visual, object-oriented extension to Promela, the input language to the Spin model checker. We introduce the v-Promela notation as supported by the VIP editor, discuss Promela code generation, and describe the process of property validation for the resulting models. Our discussion centers around two case studies, a call p...

  8. Distinguishing diffuse alopecia areata (AA) from pattern hair loss (PHL) using CD3(+) T cells.

    Science.gov (United States)

    Kolivras, Athanassios; Thompson, Curtis

    2016-05-01

    Distinguishing between diffuse subacute alopecia areata (AA), in which the peribulbar infiltrate is absent, and pattern hair loss is challenging, particularly in cases that lack marked follicular miniaturization and a marked catagen/telogen shift. We sought to distinguish diffuse AA from pattern hair loss using CD3(+) T lymphocytes. A total of 28 cases of subacute AA and 31 cases of pattern hair loss were selected and a 4-mm punch biopsy was performed. All the specimens were processed using the "HoVert" (horizontal and vertical) technique. In all cases, hematoxylin-eosin and immunohistochemical stains for CD3, CD4, CD8, and CD20 were performed. The presence of CD3(+) lymphocytes within empty follicular fibrous tracts (stela), even without a concomitant peribulbar infiltrate, is a reliable histopathological clue in supporting a diagnosis of AA (sensitivity 0.964, specificity 1, P ≤ .001). Limited tissue for analysis remained in the clinical sample tissue blocks. The presence of CD3(+) T-cells within empty follicular fibrous tracts (stela) supports a diagnosis of AA. Copyright © 2015 American Academy of Dermatology, Inc. Published by Elsevier Inc. All rights reserved.

  9. PreproVIP-derived peptides in the human female genital tract: expression and biological function

    DEFF Research Database (Denmark)

    Bredkjoer, H E; Palle, C; Ekblad, E

    1997-01-01

    The aim of the study was to elucidate the localization, distribution, colocalization and biological effect of preproVIP-derived peptides in the human female genital tract. Radioimmunoassays applying antisera against the five functional domains of the VIP precursor in combination with immunohistoc......The aim of the study was to elucidate the localization, distribution, colocalization and biological effect of preproVIP-derived peptides in the human female genital tract. Radioimmunoassays applying antisera against the five functional domains of the VIP precursor in combination...... with immunohistochemistry were used. The effect of preproVIP 22-79, preproVIP 111-122 and preproVIP 156-170 on genital smooth muscle activity in the Fallopian tube was investigated in vitro and compared to that of VIP. All the preproVIP-derived peptides were expressed throughout the genital tract in neuronal elements...

  10. Bacillus thuringiensis Cry1Ca-resistant Spodoptera exigua lacks expression of one of four Aminopeptidase N genes

    Directory of Open Access Journals (Sweden)

    Moar William J

    2005-06-01

    Full Text Available Abstract Background Insecticidal toxins from Bacillus thuringiensis bind to receptors on midgut epithelial cells of susceptible insect larvae. Aminopeptidases N (APNs from several insect species have been shown to be putative receptors for these toxins. Here we report the cloning and expression analysis of four APN cDNAs from Spodoptera exigua. Results Suppression Subtractive Hybridization (SSH was used to construct cDNA libraries of genes that are up-and down-regulated in the midgut of last instar larvae of beet armyworm, S. exigua exposed to B. thuringiensis Cry1Ca toxin. Among the clones from the SSH libraries, cDNA fragments coding for two different APNs were obtained (APN2 and APN4. A similar procedure was employed to compare mRNA differences between susceptible and Cry1Ca resistant S. exigua. Among the clones from this last comparison, cDNA fragments belonging to a third APN (APN1 were detected. Using sequences obtained from the three APN cDNA fragments and degenerate primers for a fourth APN (APN3, the full length sequences of four S. exigua APN cDNAs were obtained. Northern blot analysis of expression of the four APNs showed complete absence of APN1 expression in the resistant insects, while the other three APNs showed similar expression levels in the resistant and susceptible insects. Conclusion We have cloned and characterized four different midgut APN cDNAs from S. exigua. Expression analysis revealed the lack of expression of one of these APNs in the larvae of a Cry1Ca-resistant colony. Combined with previous evidence that shows the importance of APN in the mode of action of B. thuringiensis toxins, these results suggest that the lack of APN1 expression plays a role in the resistance to Cry1Ca in this S. exigua colony.

  11. PENGEMBANGBIAKAN Bacillus thuringiensis H-14 GALUR LOKAL PADA BERBAGAI MACAM PH MEDIA AIR KELAPA DAN TOKSISITASNYA TERHADAP JENTIK NYAMUK VEKTOR Aedes aegypti DAN Anopheles aconitus

    Directory of Open Access Journals (Sweden)

    Blodine Cb. P

    2012-12-01

    Full Text Available The culture of bioinsecticide containing active Bacillus thuringiensis H-14 local strain on various kinds on coconut water pH and its toxicity against Aedes aegypti and Anopheles aconitus were carried out in the laboratory on Institute of Vector and Reservoir Control Research and Development Salatiga. The objectives of this study were : To determine the optimum pH from various kinds of coconut water pH forculturing of B. thuringiensis H-14 local strain. This study was using 20 coconuts with 6-8 months age coconut on average weight around 1 kg that contained water approximately 400-500 ml/coconut were taken from Kunir Rejo village, Butuh regency, Purworejo district. Fifteen out of 20 coconuts were used to culture cells and spores of B. thuringiensis H-14 local strain and 5 coconuts were used to analyze the contain of coconut water in the Institute of Health Laboratory Semarang. The results showed, that B. thuringiensis H-14 local strain can culture at ranges from pH 7 to pH 8.5 with the pH7 as the optimum pH. Total Viable Cell (TVC and a Total Viable Spore Count (TVSC were 3,5 x 10 degree 10 cells /ml and 3, 3 x 10 degree 10 spores/ml respectively. The Lethal Concentration (LC50 = 10.56 ppm and LC95 = 22. 13 ppm against Ae. agypti larvae and LC50 = 5 ppm and LC95 = 11 ppm against An.aconitus larvae. The result showed the analyze test contain of coconut water were 1.92 % carbohydrate, 0.01 % fat, 0.06 % protein and reduced glucose 1.87 %. Coconut water can be used asan alternative local media to culture B. thuringiensis H-14 local strain. Key words : B. thuringiensis H-14, pH, coconut water media.

  12. Molecular analysis of two mouse dilute locus deletion mutations: Spontaneous dilute lethal20J and radiation-induced dilute prenatal lethal Aa2 alleles

    International Nuclear Information System (INIS)

    Strobel, M.C.; Seperack, P.K.; Copeland, N.G.; Jenkins, N.A.

    1990-01-01

    The dilute (d) coat color locus of mouse chromosome 9 has been identified by more than 200 spontaneous and mutagen-induced recessive mutations. With the advent of molecular probes for this locus, the molecular lesion associated with different dilute alleles can be recognized and precisely defined. In this study, two dilute mutations, dilute-lethal20J (dl20J) and dilute prenatal lethal Aa2, have been examined. Using a dilute locus genomic probe in Southern blot analysis, we detected unique restriction fragments in dl20J and Aa2 DNA. Subsequent analysis of these fragments showed that they represented deletion breakpoint fusion fragments. DNA sequence analysis of each mutation-associated deletion breakpoint fusion fragment suggests that both genomic deletions were generated by nonhomologous recombination events. The spontaneous dl20J mutation is caused by an interstitial deletion that removes a single coding exon of the dilute gene. The correlation between this discrete deletion and the expression of all dilute-associated phenotypes in dl20J homozygotes defines the dl20J mutation as a functional null allele of the dilute gene. The radiation-induced Aa2 allele is a multilocus deletion that, by complementation analysis, affects both the dilute locus and the proximal prenatal lethal-3 (pl-3) functional unit. Molecular analysis of the Aa2 deletion breakpoint fusion fragment has provided access to a previously undefined gene proximal to d. Initial characterization of this new gene suggests that it may represent the genetically defined pl-3 functional unit

  13. Cloning and Characterization of a Unique Cytotoxic Protein Parasporin-5 Produced by Bacillus thuringiensis A1100 Strain

    Directory of Open Access Journals (Sweden)

    Keisuke Ekino

    2014-06-01

    Full Text Available Parasporin is the cytocidal protein present in the parasporal inclusion of the non-insecticidal Bacillus thuringiensis strains, which has no hemolytic activity but has cytocidal activities, preferentially killing cancer cells. In this study, we characterized a cytocidal protein that belongs to this category, which was designated parasporin-5 (PS5. PS5 was purified from B. thuringiensis serovar tohokuensis strain A1100 based on its cytocidal activity against human leukemic T cells (MOLT-4. The 50% effective concentration (EC50 of PS5 to MOLT-4 cells was approximately 0.075 μg/mL. PS5 was expressed as a 33.8-kDa inactive precursor protein and exhibited cytocidal activity only when degraded by protease at the C-terminal into smaller molecules of 29.8 kDa. Although PS5 showed no significant homology with other known parasporins, a Position Specific Iterative-Basic Local Alignment Search Tool (PSI-BLAST search revealed that the protein showed slight homology to, not only some B. thuringiensis Cry toxins, but also to aerolysin-type β-pore-forming toxins (β-PFTs. The recombinant PS5 protein could be obtained as an active protein only when it was expressed in a precursor followed by processing with proteinase K. The cytotoxic activities of the protein against various mammalian cell lines were evaluated. PS5 showed strong cytocidal activity to seven of 18 mammalian cell lines tested, and low to no cytotoxicity to the others.

  14. Individual and Combined Effects of Bacillus Thuringiensis and Azadirachtin on Plodia Interpunctella Hübner (Lepidoptera: Pyralidae).

    Science.gov (United States)

    Nouri-Ganbalani, Gadir; Borzoui, Ehsan; Abdolmaleki, Arman; Abedi, Zahra; George Kamita, Shizuo

    2016-01-01

    The Indianmeal moth, Plodia interpunctella Hübner (Lepidoptera: Pyralidae), is a major stored product pest that is found throughout the world. In this study, the effect of oral exposure to Bacillus thuringiensis (Berliner) subsp. kurstaki (Bacillales: Bacillaceae) and azadirachtin was evaluated in third instar P. interpunctella under laboratory conditions. The median lethal concentration (LC50) of Bt and azadirachtin on third instars was 490 and 241 μg a.i./ml, respectively. The median lethal time (LT50) of these insecticides was the same (4.5 d following exposure to 750 or 400 μg a.i./ml of Bt or azadirachtin, respectively). When the larvae fed on diet containing LC30 concentrations of both Bt and azadirachtin an additive interaction in terms of mortality was found. A synergistic interaction was found when the larvae fed on diet containing LC50 concentrations of both insecticides. Larvae that fed on insecticide-containing diet (either Bt or azadirachtin at an LC30 concentration, or both insecticides at LC30 or LC50 concentrations) showed lower glycogen and lipid levels, and generally lower protein content in comparison to control larvae. Larvae that fed on diet containing both Bt and azadirachtin showed reduced weight gain and nutritional indices in comparison to control larvae or larvae fed on the diet containing only one of the insecticides. Finally, exposure to both insecticides, either individually or in combination, reduced the level of digestive enzymes found in the midgut. Our findings indicate that both Bt and azadirachtin, either individually or in combination have significant potential for use in controlling of P. interpunctella. © The Author 2016. Published by Oxford University Press on behalf of the Entomological Society of America.

  15. Elevated atmospheric ozone increases concentration of insecticidal Bacillus thuringiensis (Bt) Cry1Ac protein in Bt Brassica napus and reduces feeding of a Bt target herbivore on the non-transgenic parent

    International Nuclear Information System (INIS)

    Himanen, Sari J.; Nerg, Anne-Marja; Nissinen, Anne; Stewart, C. Neal; Poppy, Guy M.; Holopainen, Jarmo K.

    2009-01-01

    Sustained cultivation of Bacillus thuringiensis (Bt) transgenic crops requires stable transgene expression under variable abiotic conditions. We studied the interactions of Bt toxin production and chronic ozone exposure in Bt cry1Ac-transgenic oilseed rape and found that the insect resistance trait is robust under ozone elevations. Bt Cry1Ac concentrations were higher in the leaves of Bt oilseed rape grown under elevated ozone compared to control treatment, measured either per leaf fresh weight or per total soluble protein of leaves. The mean relative growth rate of a Bt target herbivore, Plutella xylostella L. larvae was negative on Bt plants in all ozone treatments. On the non-transgenic plants, larval feeding damage was reduced under elevated ozone. Our results indicate the need for monitoring fluctuations in Bt toxin concentrations to reveal the potential of ozone exposure for altering dosing of Bt proteins to target and non-target herbivores in field environments experiencing increasing ozone pollution. - Elevated atmospheric ozone can induce fluctuations in insecticidal protein concentrations in transgenic plants

  16. Elevated atmospheric ozone increases concentration of insecticidal Bacillus thuringiensis (Bt) Cry1Ac protein in Bt Brassica napus and reduces feeding of a Bt target herbivore on the non-transgenic parent

    Energy Technology Data Exchange (ETDEWEB)

    Himanen, Sari J. [University of Kuopio, Department of Environmental Science, P.O. Box 1627, FIN-70211 Kuopio (Finland)], E-mail: sari.himanen@uku.fi; Nerg, Anne-Marja [University of Kuopio, Department of Environmental Science, P.O. Box 1627, FIN-70211 Kuopio (Finland); Nissinen, Anne [University of Kuopio, Department of Environmental Science, P.O. Box 1627, FIN-70211 Kuopio (Finland); MTT Agrifood Research Finland, Plant Protection, FIN-31600 Jokioinen (Finland); Stewart, C. Neal [University of Tennessee, Department of Plant Sciences, Knoxville, TN 37996-4561 (United States); Poppy, Guy M. [University of Southampton, School of Biological Sciences, Southampton SO16 7PX (United Kingdom); Holopainen, Jarmo K. [University of Kuopio, Department of Environmental Science, P.O. Box 1627, FIN-70211 Kuopio (Finland)

    2009-01-15

    Sustained cultivation of Bacillus thuringiensis (Bt) transgenic crops requires stable transgene expression under variable abiotic conditions. We studied the interactions of Bt toxin production and chronic ozone exposure in Bt cry1Ac-transgenic oilseed rape and found that the insect resistance trait is robust under ozone elevations. Bt Cry1Ac concentrations were higher in the leaves of Bt oilseed rape grown under elevated ozone compared to control treatment, measured either per leaf fresh weight or per total soluble protein of leaves. The mean relative growth rate of a Bt target herbivore, Plutella xylostella L. larvae was negative on Bt plants in all ozone treatments. On the non-transgenic plants, larval feeding damage was reduced under elevated ozone. Our results indicate the need for monitoring fluctuations in Bt toxin concentrations to reveal the potential of ozone exposure for altering dosing of Bt proteins to target and non-target herbivores in field environments experiencing increasing ozone pollution. - Elevated atmospheric ozone can induce fluctuations in insecticidal protein concentrations in transgenic plants.

  17. Signaling pathways in PACAP regulation of VIP gene expression in human neuroblastoma cells

    DEFF Research Database (Denmark)

    Falktoft, B.; Georg, B.; Fahrenkrug, J.

    2009-01-01

    Ganglia expressing the neuropeptide pituitary adenylate cyclase-activating polypeptide (PACAP) innervate vasoactive intestinal peptide (VIP) containing neurons suggesting a role of PACAP in regulating VIP expression. Human NB-1 neuroblastoma cells were applied to study PACAP regulated VIP gene...... in PACAP regulation of the FOS and VIP gene expressions suggest for the first time a role of FOS in PACAP-induced VIP gene expression in human NB-1 neuroblastoma cells. (C) 2009 Elsevier Ltd. All rights reserved Udgivelsesdato: 2009/10...

  18. Interaction of PHM, PHI and 24-glutamine PHI with human VIP receptors from colonic epithelium: comparison with rat intestinal receptors

    International Nuclear Information System (INIS)

    Laburthe, M.; Couvineau, A.; Rouyer-Fessard, C.; Moroder, L.

    1985-01-01

    PHM, the human counterpart of porcine Peptide Histidine Isoleucine amide (PHI), is shown to be a VIP agonist with low potency on human VIP receptors located in colonic epithelial cell membranes. Its potency is identical to that of PHI but by 3 orders of magnitude lower than that of VIP itself in inhibiting 125 I-VIP binding and in stimulating adenylate cyclase activity. This contrasts markedly with the behavior of PHI on rat VIP receptors located in intestinal epithelial cell membranes where PHI is a potent agonist with a potency that is 1/5 that of VIP. In another connection, the authors show that 24-glutamine PHI has the same affinity as 24-glutamic acid PHI (the natural peptide) for rat or human VIP receptors. These results indicate that while PHI may exert some physiological function through its interaction with VIP receptors in rodents, its human counterpart PHM is a very poor agonist of VIP in human. Furthermore, they show that the drastic change in position 24 of PHI (neutral versus acid residue) does not affect the activity of PHI, at least on VIP receptors. 21 references, 1 figure

  19. Eco-friendly microbial route to synthesize cobalt nanoparticles using Bacillus thuringiensis against malaria and dengue vectors.

    Science.gov (United States)

    Marimuthu, Sampath; Rahuman, Abdul Abdul; Kirthi, Arivarasan Vishnu; Santhoshkumar, Thirunavukkarasu; Jayaseelan, Chidambaram; Rajakumar, Govindasamy

    2013-12-01

    The developments of resistance and persistence to chemical insecticides and concerns about the non-target effects have prompted the development of eco-friendly mosquito control agents. The aim of this study was to investigate the larvicidal activities of synthesized cobalt nanoparticles (Co NPs) using bio control agent, Bacillus thuringiensis against malaria vector, Anopheles subpictus and dengue vector, Aedes aegypti (Diptera: Culicidae). The synthesized Co NPs were characterized by X-ray diffraction (XRD), Fourier transform infrared (FTIR), Field-emission scanning electron microscopy (FESEM) with energy dispersive X-ray spectroscopy, and Transmission electron microscopy (TEM). XRD analysis showed three distinct diffraction peaks at 27.03°, 31.00°, and 45.58° indexed to the planes 102, 122, and 024, respectively on the face-centered cubic cobalt acetate with an average size of 85.3 nm. FTIR spectra implicated role of the peak at 3,436 cm(-1) for O-H hydroxyl group, 2924 cm(-1) for methylene C-H stretch in the formation of Co NPs. FESEM analysis showed the topological and morphological appearance of NPs which were found to be spherical and oval in shape. TEM analysis showed polydispersed and clustered NPs with an average size of 84.81 nm. The maximum larvicidal mortality was observed in the cobalt acetate solution, B. thuringiensis formulation, and synthesized Co NPs against fourth instar larvae of A. subpictus and A. aegypti with LC50 values of 29.16, 8.12, 3.59 mg/L; 34.61, 6.94, and 2.87 mg/L; r (2) values of 0.986, 0.933, 0.942; 0.962, 0.957, and 0.922, respectively.

  20. Using mass-release of engineered insects to manage insecticide resistance

    International Nuclear Information System (INIS)

    Alphey, Nina; Coleman, Paul G.; Donnelly, Christl A.

    2006-01-01

    Transgenic crops expressing insecticidal toxins derived from Bacillus thuringiensis (Bt) are widely used to control insect pests. The benefits of such crops would be lost if resistance to the toxins spread to a significant proportion of the pest population. The main resistance management method, mandatory in the US, is the high-dose/refuge strategy, requiring nearby refuges of toxin-free crops, and the use of toxin doses sufficiently high to kill not only wild type insects but also insects heterozygous for a resistance allele, thereby rendering the resistance functionally recessive. We propose that mass-release of harmless toxin-sensitive insects could substantially delay or even reverse the spread of resistance. Mass-release of such insects is an integral part of RIDL, a genetics-based method of pest control related to the Sterile Insect Technique. We used a population genetic mathematical model to analyze the effects of releasing male insects homozygous for a female-specific dominant lethal genetic construct, and concluded that this RIDL strategy could form an effective component of a resistance management scheme for insecticidal plants and other toxins. (author)

  1. Using mass-release of engineered insects to manage insecticide resistance

    Energy Technology Data Exchange (ETDEWEB)

    Alphey, Nina [University of Oxford (United Kingdom). Dept. of Zoology; Alphey, Luke [Oxitec Limited, Oxford (United Kingdom); Coleman, Paul G [London School of Hygiene and Tropical Medicine (United Kingdom). Dept. of Infectious and Tropical Diseases; Donnelly, Christl A [Imperial College Faculty of Medicine, London (United Kingdom). Dept. of Infectious Disease Epidemiology

    2006-07-01

    Transgenic crops expressing insecticidal toxins derived from Bacillus thuringiensis (Bt) are widely used to control insect pests. The benefits of such crops would be lost if resistance to the toxins spread to a significant proportion of the pest population. The main resistance management method, mandatory in the US, is the high-dose/refuge strategy, requiring nearby refuges of toxin-free crops, and the use of toxin doses sufficiently high to kill not only wild type insects but also insects heterozygous for a resistance allele, thereby rendering the resistance functionally recessive. We propose that mass-release of harmless toxin-sensitive insects could substantially delay or even reverse the spread of resistance. Mass-release of such insects is an integral part of RIDL, a genetics-based method of pest control related to the Sterile Insect Technique. We used a population genetic mathematical model to analyze the effects of releasing male insects homozygous for a female-specific dominant lethal genetic construct, and concluded that this RIDL strategy could form an effective component of a resistance management scheme for insecticidal plants and other toxins. (author)

  2. NCBI nr-aa BLAST: CBRC-DNOV-01-2690 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-DNOV-01-2690 ref|ZP_00743125.1| COME operon protein 3 [Bacillus thuringiensis serovar israel...ensis ATCC 35646] gb|EAO52604.1| COME operon protein 3 [Bacillus thuringiensis serovar israelensis ATCC 35646] ZP_00743125.1 0.65 31% ...

  3. Plasmid-associated sensitivity of Bacillus thuringiensis to UV light

    International Nuclear Information System (INIS)

    Benoit, T.G.; Wilson, G.R.; Bull, D.L.; Aronson, A.I.

    1990-01-01

    Spores and vegetative cells of Bacillus thuringiensis were more sensitive to UV light than were spores or cells of plasmid-cured B. thuringiensis strains or of the closely related Bacillus cereus. Introduction of B. thuringiensis plasmids into B. cereus by cell mating increased the UV sensitivity of the cells and spores. Protoxins encoded by one or more B. thuringiensis plasmids were not involved in spore sensitivity, since a B. thuringiensis strain conditional for protoxin accumulation was equally sensitive at the permissive and nonpermissive temperatures. In addition, introduction of either a cloned protoxin gene, the cloning vector, or another plasmid not containing a protoxin gene into a plasmid-cured strain of B. thuringiensis all increased the UV sensitivity of the spores. Although the variety of small, acid-soluble proteins was the same in the spores of all strains examined, the quantity of dipicolinic acid was about twice as high in the plasmid-containing strains, and this may account for the differences in UV sensitivity of the spores. The cells of some strains harboring only B. thuringiensis plasmids were much more sensitive than cells of any of the other strains, and the differences were much greater than observed with spores

  4. 40 CFR 174.505 - Bacillus thuringiensis modified Cry3A protein (mCry3A) in corn; exemption from the requirement of...

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 23 2010-07-01 2010-07-01 false Bacillus thuringiensis modified Cry3A protein (mCry3A) in corn; exemption from the requirement of a tolerance. 174.505 Section 174.505 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) PESTICIDE PROGRAMS PROCEDURES AND REQUIREMENTS FOR PLANT-INCORPORATED PROTECTANTS...

  5. Variation in VIP latrine sludge contents | Bakare | Water SA

    African Journals Online (AJOL)

    This study investigated variations in the characteristics of the sludge content from different ventilated improved pit (VIP) latrines and variation in these characteristics at specific depths within each pit. Faecal sludge from 16 VIP latrines within the eThekwini Municipality was collected and laboratory characterisation including ...

  6. Expression and characterization of preproVIP derived peptides in the human male urogenital tract

    DEFF Research Database (Denmark)

    Ottesen, B; Bredkjaer, H E; Ekblad, E

    1995-01-01

    Expression of the gene sequence encoding vasoactive intestinal polypeptide (VIP) leads to the synthesis of a 170 amino acid precursor molecule which can be processed to five fragments: preproVIP 22-79, peptide histidine methionine (PHM), or peptide histidine valine (PHV), preproVIP 111-122, VIP...

  7. Occurrence of Bacillus thuringiensis in faeces of herbivorous farm ...

    African Journals Online (AJOL)

    Bacillus thuringiensis (Berliner), the insect pathogen has been isolated from a variety of habitat. It is understood that the habitat of B. thuringiensis has always been associated with their biological activity. In the present study, B. thuringiensis was isolated from faeces of cows and goats. The phenotypic characterization ...

  8. Virtual phantom magnetic resonance imaging (ViP MRI) on a clinical MRI platform.

    Science.gov (United States)

    Saint-Jalmes, Hervé; Bordelois, Alejandro; Gambarota, Giulio

    2018-01-01

    The purpose of this study was to implement Virtual Phantom Magnetic Resonance Imaging (ViP MRI), a technique that allows for generating reference signals in MR images using radiofrequency (RF) signals, on a clinical MR system and to test newly designed virtual phantoms. MRI experiments were conducted on a 1.5 T MRI scanner. Electromagnetic modelling of the ViP system was done using the principle of reciprocity. The ViP RF signals were generated using a compact waveform generator (dimensions of 26 cm × 18 cm × 16 cm), connected to a homebuilt 25 mm-diameter RF coil. The ViP RF signals were transmitted to the MRI scanner bore, simultaneously with the acquisition of the signal from the object of interest. Different types of MRI data acquisition (2D and 3D gradient-echo) as well as different phantoms, including the Shepp-Logan phantom, were tested. Furthermore, a uniquely designed virtual phantom - in the shape of a grid - was generated; this newly proposed phantom allows for the investigations of the vendor distortion correction field. High quality MR images of virtual phantoms were obtained. An excellent agreement was found between the experimental data and the inverse cube law, which was the expected functional dependence obtained from the electromagnetic modelling of the ViP system. Short-term time stability measurements yielded a coefficient of variation in the signal intensity over time equal to 0.23% and 0.13% for virtual and physical phantom, respectively. MR images of the virtual grid-shaped phantom were reconstructed with the vendor distortion correction; this allowed for a direct visualization of the vendor distortion correction field. Furthermore, as expected from the electromagnetic modelling of the ViP system, a very compact coil (diameter ~ cm) and very small currents (intensity ~ mA) were sufficient to generate a signal comparable to that of physical phantoms in MRI experiments. The ViP MRI technique was successfully implemented on a clinical MR

  9. A Fungal Insecticide Engineered for Fast Per Os Killing of Caterpillars Has High Field Efficacy and Safety in Full-Season Control of Cabbage Insect Pests

    Science.gov (United States)

    Liu, Yong-Jie; Liu, Jing; Ying, Sheng-Hua; Liu, Shu-Sheng

    2013-01-01

    Fungal insecticides developed from filamentous pathogens of insects are notorious for their slow killing action through cuticle penetration, depressing commercial interest and practical application. Genetic engineering may accelerate their killing action but cause ecological risk. Here we show that a Beauveria bassiana formulation, HV8 (BbHV8), engineered for fast per os killing of caterpillars by an insect midgut-acting toxin (Vip3Aa1) overexpressed in conidia has both high field efficacy and safety in full-season protection of cabbage from the damage of an insect pest complex dominated by Pieris rapae larvae, followed by Plutella xylostella larvae and aphids. In two fields repeatedly sprayed during summer, BbHV8 resulted in overall mean efficacies of killing of 71% and 75%, which were similar or close to the 70% and 83% efficacies achieved by commercially recommended emamectin benzoate but much higher than the 31% and 48% efficacies achieved by the same formulation of the parental wild-type strain (WT). Both BbHV8 and WT sprays exerted no adverse effect on a nontarget spider community during the trials, and the sprays did not influence saprophytic fungi in soil samples taken from the field plots during 4 months after the last spray. Strikingly, BbHV8 and the WT showed low fitness when they were released into the environment because both were decreasingly recovered from the field lacking native B. bassiana strains (undetectable 5 months after the spray), and the recovered isolates became much less tolerant to high temperature and UV-B irradiation. Our results highlight for the first time that a rationally engineered fungal insecticide can compete with a chemical counterpart to combat insect pests at an affordable cost and with low ecological risk. PMID:23956386

  10. The adsorption features between insecticidal crystal protein and nano-Mg(OH)2.

    Science.gov (United States)

    Pan, Xiaohong; Xu, Zhangyan; Zheng, Yilin; Huang, Tengzhou; Li, Lan; Chen, Zhi; Rao, Wenhua; Chen, Saili; Hong, Xianxian; Guan, Xiong

    2017-12-01

    Nano-Mg(OH) 2 , with low biological toxicity, is an ideal nano-carrier for insecticidal protein to improve the bioactivity. In this work, the adsorption features of insecticidal protein by nano-Mg(OH) 2 have been studied. The adsorption capacity could reach as high as 136 mg g -1 , and the adsorption isotherm had been fitted with Langmuir and Freundlich models. Moreover, the adsorption kinetics followed a pseudo-first or -second order rate model, and the adsorption was spontaneous and an exothermic process. However, high temperatures are not suitable for adsorption, which implies that the temperature would be a critical factor during the adsorption process. In addition, FT-IR confirmed that the protein was adsorbed on the nano-Mg(OH) 2 , zeta potential analysis suggested that insecticidal protein was loaded onto the nano-Mg(OH) 2 not by electrostatic adsorption but maybe by intermolecular forces, and circular dichroism spectroscopy of Cry11Aa protein before and after loading with nano-Mg(OH) 2 was changed. The study applied the adsorption information between Cry11Aa and nano-Mg(OH) 2 , which would be useful in the practical application of nano-Mg(OH) 2 as a nano-carrier.

  11. Insect Ryanodine Receptor: Distinct But Coupled Insecticide Binding Sites for [N-C3H3]Chlorantraniliprole, Flubendiamide, and [3H]Ryanodine

    OpenAIRE

    Isaacs, André K.; Qi, Suzhen; Sarpong, Richmond; Casida, John E.

    2012-01-01

    Radiolabeled anthranilic diamide insecticide [N-C3H3]chlorantraniliprole was synthesized at high specific activity and compared with phthalic diamide insecticide flubendiamide and [3H]ryanodine in radioligand binding studies with house fly muscle membranes to provide the first direct evidence with a native insect ryanodine receptor that the major anthranilic and phthalic diamide insecticides bind at different allosterically coupled sites, i.e. there are three distinct Ca2+-release channel tar...

  12. An overview of the safety and biological effects of Bacillus thuringiensis Cry toxins in mammals.

    Science.gov (United States)

    Rubio-Infante, Néstor; Moreno-Fierros, Leticia

    2016-05-01

    Crystal proteins (Cry) produced during the growth and sporulation phases of Bacillus thuringiensis (Bt) bacterium are known as delta endotoxins. These toxins are being used worldwide as bioinsecticides to control pests in agriculture, and some Cry toxins are used against mosquitoes to control vector transmission. This review summarizes the relevant information currently available regarding the biosafety and biological effects that Bt and its insecticidal Cry proteins elicit in mammals. This work was performed because of concerns regarding the possible health impact of Cry toxins on vertebrates, particularly because Bt toxins might be associated with immune-activating or allergic responses. The controversial data published to date are discussed in this review considering earlier toxicological studies of B. thuringiensis, spores, toxins and Bt crops. We discussed the experimental studies performed in humans, mice, rats and sheep as well as in diverse mammalian cell lines. Although the term 'toxic' is not appropriate for defining the effects these toxins have on mammals, they cannot be considered innocuous, as they have some physiological effects that may become pathological; thus, trials that are more comprehensive are necessary to determine their effects on mammals because knowledge in this field remains limited. Copyright © 2015 John Wiley & Sons, Ltd.

  13. Controle de tortricídeos em macieira com duas formulações de Bacillus thuringiensis var. Kustaki em Fraiburgo-SC Tortricid moth control in apple with two formulation of Bacillus thuringiensis var. Kustaki in Fraiburgo-SC

    Directory of Open Access Journals (Sweden)

    Lino Bittencourt Monteiro

    2010-06-01

    Full Text Available Grapholita molesta e Bonagota cranaodes são duas importantes pragas de pomares de macieira controlada por inseticidas. O objetivo deste estudo foi testar duas formulações de Bacillus thuringiensis var. kurstaki (Btk para o controle desses tortricídeos. Parcelas de macieira 'Fuji' foram pulverizadas com Dipel PM e Dipel SC, nas concentrações de 100 ml por100 L de calda. A eficiência do Btk foi comparada com os inseticidas tebufenozide (Mimic 240 SC - 90 ml por 100 L e clorpyrifos (Lorsban 480 BR - 150 ml por 100 L. Duas vezes por semana, eram avaliadas as capturas de pragas em armadilha do tipo Delta. As avaliações de danos nos frutos foram realizadas antes e na colheita, sendo classificados os frutos em função do agrupamento em cachopa e a sua localização na planta. Os tratamentos com Btk tiveram mais danos do que os químicos. Houve uma tendência de maiores danos na parte interna, próximo ao tronco das macieiras e o agrupamento dos frutos não influenciou na presença de danos. Ambas as formulações de Dipel foram eficientes quando comparados com Mimic e Lorsban.Grapholita molesta and Bonagota cranaodes are two key pests in apple orchards controlled by insecticides. The objective of the present study was to test two formulas of Bacillus thuringiensis var. kurstaki to control these tortricid moths. Plot of 'Fuji' apple trees were sprayed with Dipel PM and Dipel SC, at a concentration of 100ml por 100 l liquid. The efficiency of Btk was compared with the insecticides tebuphenozide (Mimic 240 SC - 90 ml por 100 L and chlorpyrifos (Lorsban 480 BR - 150 ml por 100 L. The capture of pests in a Delta-type trap was assessed twice a week. Fruit damage was evaluated pre- and post- harvest and the fruit were classified by branch-end clustering and location on the plant. The treatments with B. thuringiensis were more damaged than those with chemical treatments. There tended to be more damage in the internal part, close to the apple tree

  14. NCBI nr-aa BLAST: CBRC-CJAC-01-0048 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-CJAC-01-0048 ref|NP_054012.1| collagen-like protein [Bacillus thuringiensis serovar israel...ensis] gb|AAB16964.1| collagen-like protein [Bacillus thuringiensis serovar israelensis] NP_054012.1 3e-11 36% ...

  15. NCBI nr-aa BLAST: CBRC-CJAC-01-1058 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-CJAC-01-1058 ref|NP_054012.1| collagen-like protein [Bacillus thuringiensis serovar israel...ensis] gb|AAB16964.1| collagen-like protein [Bacillus thuringiensis serovar israelensis] NP_054012.1 3e-38 39% ...

  16. Structure-activity studies of vasoactive intestinal peptide (VIP): cyclic disulfide analogs.

    Science.gov (United States)

    Bolin, D R; Cottrell, J; Garippa, R; O'Neill, N; Simko, B; O'Donnell, M

    1993-02-01

    Analogs of vasoactive intestinal peptide with cysteine residues incorporated at selected sites within the sequence were prepared by solid phase methods, oxidized to the corresponding cyclic disulfides and purified to homogeneity by preparative HPLC. The cyclic compounds were assayed as smooth muscle relaxants on isolated guinea pig trachea, as bronchodilators in vivo in guinea pigs, and for binding to VIP receptors in guinea pig lung membranes. Of the analogs prepared at the N-terminus, one compound, Ac-[D-Cys6,D-Cys11,Lys12,Nle17,Val26,Th r28]-VIP, was found to be a full agonist with slightly more than one tenth the potency of native VIP. Most other cyclic analogs in the N-terminal region were found to be inactive. A second analog, Ac-[Lys12,Cys17,Val26,Cys28]-VIP, was also found to be a full agonist with potency about one third that of native VIP. Furthermore, this compound was active as a bronchodilator in vivo in guinea pig, but with somewhat diminished potency as compared to native VIP. Strikingly, this cyclic compound was found to have significantly longer duration of action (> 40 min) when compared to an analogous acyclic compound (5 min). The conformational restrictions imposed by formation of the cyclic ring structures may have stabilized the molecule to degradation, thus enhancing the effective duration of action. Analysis of this series of cyclic analogs has also yielded information about the requirements for the receptor-active conformation of VIP.

  17. Fundamental and clinical evaluation of vasoactive intestinal peptide (VIP) in pancreatitis by radioimmunoassay kit

    International Nuclear Information System (INIS)

    Uehara, Soichiro; Nakamura, Takayuki; Ihara, Kenichi; Isogawa, Shin; Hirayama, Akio

    1987-01-01

    Plasma vasoactive intestinal peptide (VIP) concentrations of normal individuals and patients with pancreatitis were studied using a VIP RIA kit. The inter-assay and intra-assay variation of this kit were between 2.1 and 9.4 %. The VIP levels increased in the acute phase of acute pancreatitis and patients with chronic pancreatitis. The VIP concentration increased during the first 30 min of glucose tolerance test, but this increase was much smaller than that in insulin. These results suggest that this kit is useful for physiologic and pathologic changes in the VIP level. (author)

  18. Ontogeny of the VIP system in the gastro-intestinal tract of the Axolotl, Ambystoma mexicanum: successive appearance of co-existing PACAP and NOS.

    Science.gov (United States)

    Badawy, Gamal; Reinecke, Manfred

    2003-03-01

    Evidence for the presence and potential co-existence of vasoactive intestinal polypeptide (VIP), pituitary adenylate cyclase-activating polypeptide (PACAP) and nitric oxide synthase (NOS) in gastro-intestinal endocrine cells and/or nerve fibers is conflicting and very few results exist on development. This immunofluorescence study aims to clarify the appearance and localization of VIP, PACAP and NOS in the gastro-intestinal tract of the Axolotl, Ambystoma mexicanum, during ontogeny. VIP-immunoreactivity appeared in nerve fibers as early as on day 3 after hatching likely indicating a particular role, such as a trophic action, of VIP in very early development. PACAP-immunoreactivity was observed 3 days later within the VIP-immunoreactive (-IR) fibers. From this time on, VIP- and PACAP-immunoreactivity exhibited complete co-existence. VIP/PACAP-IR fibers were found throughout the gastro-intestinal tract. They were most prominent in the myenteric plexus and the muscle layers and less frequent in the submucosa. NOS-immunoreactivity appeared as late as at the 1st (64 days) juvenile stage in a subpopulation of the VIP/PACAP-IR fibers that contacted submucosal arteries. We found only very few VIP/PACAP-IR perikarya, indicating that part of the VIP/PACAP-IR fibers is of extrinsic origin. On day 12 and in the 1st and 2nd (104 days) juvenile stage, infrequent PACAP-IR entero-endocrine cells were noted, while neither VIP- nor NOS-immunoreactivity occurred in endocrine cells at any stage of development. The complete coexistence of neuronal PACAP- and VIP-immunoreactivities and their very early appearance in ontogeny may suggest important and coordinated roles of both peptides in the control of Axolotl gastro-intestinal activity, while the VIP/ PACAP/NOS-IR fibers may be involved in the regulation of submucosal blood flow.

  19. Chemical and thermal properties of VIP latrine sludge

    African Journals Online (AJOL)

    2015-07-04

    Jul 4, 2015 ... This study investigated the chemical and thermal properties of faecal sludge from 10 dry VIP latrines in Bester's Camp in the eThekwini Municipality, Durban, ... emptying and treatment equipment. A manual sorting of the pit .... (LaDePa) plant (Harrison and Wilson, 2012). Figure 3 illustrates the depths of the ...

  20. Binding site alteration is responsible for field-isolated resistance to Bacillus thuringiensis Cry2A insecticidal proteins in two Helicoverpa species.

    Directory of Open Access Journals (Sweden)

    Silvia Caccia

    Full Text Available BACKGROUND: Evolution of resistance by target pests is the main threat to the long-term efficacy of crops expressing Bacillus thuringiensis (Bt insecticidal proteins. Cry2 proteins play a pivotal role in current Bt spray formulations and transgenic crops and they complement Cry1A proteins because of their different mode of action. Their presence is critical in the control of those lepidopteran species, such as Helicoverpa spp., which are not highly susceptible to Cry1A proteins. In Australia, a transgenic variety of cotton expressing Cry1Ac and Cry2Ab (Bollgard II comprises at least 80% of the total cotton area. Prior to the widespread adoption of Bollgard II, the frequency of alleles conferring resistance to Cry2Ab in field populations of Helicoverpa armigera and Helicoverpa punctigera was significantly higher than anticipated. Colonies established from survivors of F(2 screens against Cry2Ab are highly resistant to this toxin, but susceptible to Cry1Ac. METHODOLOGY/PRINCIPAL FINDINGS: Bioassays performed with surface-treated artificial diet on neonates of H. armigera and H. punctigera showed that Cry2Ab resistant insects were cross-resistant to Cry2Ae while susceptible to Cry1Ab. Binding analyses with (125I-labeled Cry2Ab were performed with brush border membrane vesicles from midguts of Cry2Ab susceptible and resistant insects. The results of the binding analyses correlated with bioassay data and demonstrated that resistant insects exhibited greatly reduced binding of Cry2Ab toxin to midgut receptors, whereas no change in (125I-labeled-Cry1Ac binding was detected. As previously demonstrated for H. armigera, Cry2Ab binding sites in H. punctigera were shown to be shared by Cry2Ae, which explains why an alteration of the shared binding site would lead to cross-resistance between the two Cry2A toxins. CONCLUSION/SIGNIFICANCE: This is the first time that a mechanism of resistance to the Cry2 class of insecticidal proteins has been reported

  1. High sensitivity tests of the Pauli Exclusion Principle with VIP2

    CERN Document Server

    Marton, J; Bertolucci, S; Berucci, C; Bragadireanu, M; Cargnelli, M; Curceanu, C; Clozza, A; Di Matteo, S; Egger, J-P; Guaraldo, C; Iliescu, M; Ishiwatari, T; Laubenstein, M; Milotti, E; Pichler, A; Pietreanu, D; Piscicchia, K; Ponta, T; Scordo, A; Shi, H; Sirghi, D L; Sirghi, F; Sperandio, L; Doce, O Vazquez; Widmann, E; Zmeskal, J

    2015-01-01

    The Pauli Exclusion Principle is one of the most fundamental rules of nature and represents a pillar of modern physics. According to many observations the Pauli Exclusion Principle must be extremely well fulfilled. Nevertheless, numerous experimental investigations were performed to search for a small violation of this principle. The VIP experiment at the Gran Sasso underground laboratory searched for Pauli-forbidden X-ray transitions in copper atoms using the Ramberg-Snow method and obtained the best limit so far. The follow-up experiment VIP2 is designed to reach even higher sensitivity. It aims to improve the limit by VIP by orders of magnitude. The experimental method, comparison of different PEP tests based on different assumptions and the developments for VIP2 are presented.

  2. NCBI nr-aa BLAST: CBRC-XTRO-01-3892 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-XTRO-01-3892 ref|ZP_00743008.1| Fiber protein [Bacillus thuringiensis serovar israel...ensis ATCC 35646] gb|EAO52717.1| Fiber protein [Bacillus thuringiensis serovar israelensis ATCC 35646] ZP_00743008.1 3e-15 42% ...

  3. 40 CFR 174.509 - Bacillus thuringiensis Cry3A protein; exemption from the requirement of a tolerance.

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 23 2010-07-01 2010-07-01 false Bacillus thuringiensis Cry3A protein; exemption from the requirement of a tolerance. 174.509 Section 174.509 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) PESTICIDE PROGRAMS PROCEDURES AND REQUIREMENTS FOR PLANT-INCORPORATED PROTECTANTS Tolerances and Tolerance...

  4. Macrophage Resistance to HIV-1 Infection Is Enhanced by the Neuropeptides VIP and PACAP

    Science.gov (United States)

    Temerozo, Jairo R.; Joaquim, Rafael; Regis, Eduardo G.; Savino, Wilson; Bou-Habib, Dumith Chequer

    2013-01-01

    It is well established that host factors can modulate HIV-1 replication in macrophages, critical cells in the pathogenesis of HIV-1 infection due to their ability to continuously produce virus. The neuropeptides VIP and PACAP induce well-characterized effects on macrophages through binding to the G protein-coupled receptors VPAC1, VPAC2 and PAC1, but their influence on HIV-1 production by these cells has not been established. Here, we describe that VIP and PACAP reduce macrophage production of HIV-1, acting in a synergistic or additive manner to decrease viral growth. Using receptor antagonists, we detected that the HIV-1 inhibition promoted by VIP is dependent on its ligation to VPAC1/2, whereas PACAP decreases HIV-1 growth via activation of the VPAC1/2 and PAC1 receptors. Specific agonists of VPAC2 or PAC1 decrease macrophage production of HIV-1, whereas sole activation of VPAC1 enhances viral growth. However, the combination of specific agonists mimicking the receptor preference of the natural neuropeptides reproduces the ability of VIP and PACAP to increase macrophage resistance to HIV-1 replication. VIP and PACAP up-regulated macrophage secretion of the β-chemokines CCL3 and CCL5 and the cytokine IL-10, whose neutralization reversed the neuropeptide-induced inhibition of HIV-1 replication. Our results suggest that VIP and PACAP and the receptors VPAC2 and PAC1 could be used as targets for developing alternative therapeutic strategies for HIV-1 infection. PMID:23818986

  5. VIP/PACAP receptors in cerebral arteries of rat

    DEFF Research Database (Denmark)

    Erdling, André; Sheykhzade, Majid; Maddahi, Aida

    2013-01-01

    BACKGROUND: Vasoactive intestinal peptide (VIP) and pituitary adenylate cyclase activating peptide (PACAP)-containing nerves surround cerebral blood vessels. The peptides have potent vasodilator properties via smooth muscle cell receptors and activation of adenylate cyclase. The purpose of this s......BACKGROUND: Vasoactive intestinal peptide (VIP) and pituitary adenylate cyclase activating peptide (PACAP)-containing nerves surround cerebral blood vessels. The peptides have potent vasodilator properties via smooth muscle cell receptors and activation of adenylate cyclase. The purpose...

  6. NCBI nr-aa BLAST: CBRC-SARA-01-0256 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-SARA-01-0256 ref|ZP_00739965.1| Integral membrane protein [Bacillus thuringiensis serovar israel...ensis ATCC 35646] gb|EAO55770.1| Integral membrane protein [Bacillus thuringiensis serovar israelensis ATCC 35646] ZP_00739965.1 1.3 24% ...

  7. VIP Gene Deletion in Mice Causes Cardiomyopathy Associated with Upregulation of Heart Failure Genes

    Energy Technology Data Exchange (ETDEWEB)

    Szema, Anthony M.; Hamidi, Sayyed A.; Smith, S. David; Benveniste, Helene; Katare, Rajesh Gopalrao

    2013-05-20

    Vasoactive Intestinal Peptide (VIP), a pulmonary vasodilator and inhibitor of vascular smooth muscle proliferation, is absent in pulmonary arteries of patients with idiopathic pulmonary arterial hypertension (PAH). We previously determined that targeted deletion of the VIP gene in mice leads to PAH with pulmonary vascular remodeling and right ventricular (RV) dilatation. Whether the left ventricle is also affected by VIP gene deletion is unknown. In the current study, we examined if VIP knockout mice (VIP-/-) develop both right (RV) and left ventricular (LV) cardiomyopathy, manifested by LV dilatation and systolic dysfunction, as well as overexpression of genes conducive to heart failure.

  8. The VIP/VPACR system in the reproductive cycle of male lizard Podarcis sicula.

    Science.gov (United States)

    Agnese, Marisa; Rosati, Luigi; Prisco, Marina; Coraggio, Francesca; Valiante, Salvatore; Scudiero, Rosaria; Laforgia, Vincenza; Andreuccetti, Piero

    2014-09-01

    Starting from the knowledge that in the reproductive period the Vasoactive Intestinal Peptide (VIP) is widely distributed in Podarcis sicula testis, we studied VIP expression and the localization of the neuropeptide and its receptors in the testis of the Italian wall lizard P. sicula in the other phases of its reproductive cycle (summer stasis, autumnal resumption, winter stasis, spring resumption). By Real Time-PCR, we demonstrated that testicular VIP mRNA levels change during the reproductive cycle, showing a cyclic trend with two peaks, one in the mid-autumnal resumption and the other in the reproductive period. By in situ hybridization and immunohistochemistry, we demonstrated that both VIP mRNA and protein were widely distributed in the testis in almost all the phases of the cycle, except in the early autumnal resumption. As regards the receptors, the VPAC1R was localized mainly in Leydig cells, while the VPAC2R showed the same distribution of VIP. Our results demonstrate that, differently from mammals, where VIP is present only in nerve fibres innerving the testis, an endotesticular synthesis takes place in the lizard and the VIP synthesis changes throughout the reproductive cycle. Moreover, the VIP/VPAC receptor system distribution observed in germ and somatic cells in various phases of the cycle, and particularly in the autumnal resumption and the reproductive period, strongly suggests its involvement in both spermatogenesis and steroidogenesis. Finally, the wider distribution of VIP in lizards with respect to mammals leads us to hypothesize that during the evolution the synthesis sites have been transferred from the testis to other districts, such as the brain. Copyright © 2014 Elsevier Inc. All rights reserved.

  9. Three-dimensional (3D) structure prediction and function analysis of the chitin-binding domain 3 protein HD73_3189 from Bacillus thuringiensis HD73.

    Science.gov (United States)

    Zhan, Yiling; Guo, Shuyuan

    2015-01-01

    Bacillus thuringiensis (Bt) is capable of producing a chitin-binding protein believed to be functionally important to bacteria during the stationary phase of its growth cycle. In this paper, the chitin-binding domain 3 protein HD73_3189 from B. thuringiensis has been analyzed by computer technology. Primary and secondary structural analyses demonstrated that HD73_3189 is negatively charged and contains several α-helices, aperiodical coils and β-strands. Domain and motif analyses revealed that HD73_3189 contains a signal peptide, an N-terminal chitin binding 3 domains, two copies of a fibronectin-like domain 3 and a C-terminal carbohydrate binding domain classified as CBM_5_12. Moreover, analysis predicted the protein's associated localization site to be the cell wall. Ligand site prediction determined that amino acid residues GLU-312, TRP-334, ILE-341 and VAL-382 exposed on the surface of the target protein exhibit polar interactions with the substrate.

  10. Iodinated derivatives of vasoactive intestinal peptide (VIP), PHI and PHM: purification, chemical characterization and biological activity

    International Nuclear Information System (INIS)

    McMaster, D.; Suzuki, Y.; Rorstad, O.; Lederis, K.

    1987-01-01

    The iodination of vasoactive intestinal peptide (VIP) was studied, using a variety of enzymatic and chemical iodination methods. Reversed phase high performance liquid chromatography (HPLC) was used to purify the reaction products. The lactoperoxidase-glucose oxidase method gave excellent results in terms of reproducibility, iodine incorporation, and yield of the non-oxidized products [Tyr(I)10]VIP and [Tyr(I)22]VIP, and was used to prepare both 125 I and 127 I labelled derivatives. In both cases, direct application to HPLC and a single column system were used. Although the oxidized peptides [Tyr(I)10,Met(O)17]VIP and [Tyr(I)22,Met(O)17]VIP could be generated to varying degrees directly by iodination of VIP, these were most conveniently prepared by iodination of [Met(O)17]VIP. Iodinated derivatives of the homologous peptides PHI and PHM were likewise prepared by rapid, one-step HPLC procedures. The site and degree of iodination were determined by HPLC peptide mapping of tryptic digests and amino acid analyses, and in the case of [Tyr(I)10]VIP also by sequencing. The vasorelaxant activities of the iodinated peptides in bovine cerebral artery preparations did not differ significantly from those of the corresponding noniodinated peptides, with the exception of [Tyr(I)10,Met(O)17]VIP and [Tyr(I)22,Met(O)17]VIP which, unlike [Met(O)17]VIP itself, had slightly lower potency than VIP

  11. Vasoactive intestinal peptide (VIP) binds to guinea pig peritoneal eosinophils: A single class of binding sites with low affinity and high capacity

    International Nuclear Information System (INIS)

    Sakakibara, H.; Shima, K.; Takamatsu, J.; Said, S.I.

    1990-01-01

    VIP binds to specific receptors on lymphocytes and mononuclear cells and exhibits antiinflammatory properties. Eosinophils (Eos) contribute to inflammatory reactions but the regulation of Eos function is incompletely understood. The authors examined the binding of monoradioiodinated VIP, [Tyr( 125 I) 10 ] VIP ( 125 I-VIP), to Eos in guinea pigs. The interaction of 125 i-VIP with Eos was rapid, reversible, saturable and linearly dependent on the number of cells. At equilibrium the binding was competitively inhibited by native peptide or by the related peptide helodermin. Scatchard analysis suggested the presence of a single class of VIP binding sites with a low affinity and a high capacity. In the presence of isobutyl-methylxanthine, VIP, PHI or helodermin did not stimulate cyclic AMP accumulation in intact Eos, while PGE 2 or 1-isoproterenol did. VIP also did not inhibit superoxide anion generation from Eos stimulated by phorbol myristate acetate. The authors conclude that: (1) VIP binds to low-affinity, specific sites on guinea pig peritoneal eosinophils; (2) this binding is not coupled to stimulation of adenylate cyclase; and (3) the possible function of these binding sites is at present unknown

  12. 3 Insecticide Use Practice

    African Journals Online (AJOL)

    Administrator

    500,000 metric tonnes in the 1964/1965 season. Problems ... insecticides on the open market. ... effective in the management of insect pests of cocoa. .... Effectiveness and profitability of pest ... Youth in Agriculture; Programme Policy, Strategy.

  13. Vasoactive intestinal peptide (VIP) labelling with iodine-131 by direct method; Marcacao do peptideo intestinal vasoativo (VIP) com iodo-131 por metodo direto

    Energy Technology Data Exchange (ETDEWEB)

    Colturato, M.T.; Silva, C.P.G. da; Araujo, E.B. [Instituto de Pesquisas Energeticas e Nucleares (IPEN), Sao Paulo, SP (Brazil). Centro de Radiofarmacia

    2002-07-01

    The Vasoactive Intestinal Peptide (VIP) is a 28-amino acid polypeptide with a great numbers of receptors in tumoral cells, including adenocarcinomas and pancreatic and colon carcinomas. The VIP molecule contains two tyrosine residues, in positions 10 and 22, that are theoretically equally susceptible to iodination, The VIP was labeled with 131-iodine by direct method using Iodogen as oxidant agent: 15.03 mmol VIP + 0.10 nmol KI + [{sup 131} I]NaI + 13.9 mmol Iodogen; the final volume was adjust to 100 {mu}L using 0.2 M phosphate buffer, pH 7.5 and the reaction proceed with stirring for 30 minutes at room temperature. The radiochemical purity was determined by electrophoresis (Whatman 1MM paper; 0.05 M barbital buffer; pH 8.6; 150 V; 40 minutes) that indicates low percent of free 131-iodine. The high performance liquid chromatography (HPLC) system using RPC{sup 18}, 10 {mu}m, 4 x 250mm column, was able to separate the different radiochemical species, only when an isocratic mixture of acetonitrile: 0.1% trifluoroacetic acid (27:73) was used, with 0.5 mL/min. flux. (author)

  14. Activity of Selected Formulated Biorational and Synthetic Insecticides Against Larvae of Helicoverpa armigera (Lepidoptera: Noctuidae).

    Science.gov (United States)

    Vivan, L M; Torres, J B; Fernandes, P L S

    2017-02-01

    This work studied 17 insecticides belonging to nucleopolyhedrovirus (NPV), Bacillus thuringiensis (Bt kurstaki and Bt aizawai), benzoylureas (insect growth regulators [IGRs]), carbamates, organophosphates, spinosyns, and diamides against larvae of Helicoverpa armigera (Hübner), invasive species in the South American continent. Larvae of different instars were fed for 7 d with untreated or insecticide-treated diets. Mortality was recorded daily for 7 d, and surviving larvae were individually weighed on the seventh day. The NPV and Bt insecticides caused 100% mortality of first-instar larvae and first-instar and second-instar larvae, respectively. However, both NPV and Bt-based products caused low mortality of third-instar larvae and did not kill older larvae. The IGR lufenuron was highly effective against all three ages of larvae tested, whereas teflubenzuron and triflumuron produced maximum 60% mortality of second-instar larvae and lower than 50% to older larvae. Thiodicarb, chlorantraniliprole, indoxacarb, chlorpyrifos, and chlorfenapyr, irrespective of tested age, caused 100% mortality of larvae, with the last two insecticides reaching 100% mortality within 2 d of feeding on the treated diet. Flubendiamide caused lower mortality but significantly affected the weight of surviving larvae, whereas neither spinosad nor methomyl produced significant mortality or affected the weight of larvae. Based on the results, the age of H. armigera larvae plays an important role in the recommendation of NPV and Bt insecticides. Furthermore, there are potential options between biological and synthetic insecticides tested against H. armigera, and recording larval size during monitoring, in addition to the infestation level, should be considered when recommending biological-based insecticides to control this pest. © The Authors 2016. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  15. Acute, sublethal and combination effects of azadirachtin and Bacillus thuringiensis toxins on Helicoverpa armigera (Lepidoptera: Noctuidae) larvae.

    Science.gov (United States)

    Singh, G; Rup, P J; Koul, Opender

    2007-08-01

    The efficacy of neem (1500 ppm azadirachtin (AI)), Delfin WG, a biological insecticide based on selected strain of Bacillus thuringiensis Berliner (Bt) subspecies kurstaki, and Cry1Ac protein, either individually or in combination, were examined against first to fourth instar Helicoverpa armigera (Hübner) larvae. Using an oral administration method, various growth inhibitory concentrations (EC) and lethal concentrations (LC) were determined for each bioagent. Combinations of sublethal concentrations of Bt spray formulation with azadirachtin at EC50 or EC95 levels not only enhanced the toxicity, but also reduced the duration of action when used in a mixture. The LC20 and LC50 values for Cry1Ac toxin were 0.06 and 0.22 microg ml-1, respectively. Bt-azadirachtin combinations of LC50+EC20 and LC50+EC50 result in 100% mortality. The mortality also was significant in LC20+EC20 and LC20+EC50 mixtures. These studies imply that the combined action is not synergistic but complimentary, with azadirachtin particularly facilitating the action of Bt. The Bt spray-azadirachtin combination is more economical than combinations that involve isolating the toxic protein, as the Bt spray formulations can be combined in a spray mixture with neem. These combinations may be useful for controlling bollworm populations that have acquired resistance to Bt as they may not survive the effect of mixture. Azadirachtin may be useful as a means of reducing the endotoxin concentrations in a mixture, to promote increased economic savings and further reduce the probability of resistance development to either insect control agent.

  16. Scorpion neurotoxin AaIT-expressing Beauveria bassiana enhances the virulence against Aedes albopictus mosquitoes.

    Science.gov (United States)

    Deng, Sheng-Qun; Cai, Qun-Di; Deng, Ming-Zhi; Huang, Qiang; Peng, Hong-Juan

    2017-12-01

    To improve the insecticidal efficacy of this entomopathogen Beauveria bassiana, the fungus was genetically modified to express an insect-specific scorpion neurotoxin AaIT. The virulence of the recombinant B. bassiana strain (Bb-AaIT) against Aedes albopictus adults (which occurs via penetration through the cuticle during spore germination or by conidia ingestion), and the larvae (by conidia ingestion) was measured with bioassays. The median lethal concentration (LC 50 ) of Bb-AaIT against A. albopictus larvae was 313.3-fold lower on day 4 and 11.3-fold lower on day 10 than that of the wild type (WT). Through conidia feeding or body contact, Bb-AaIT killed 50% of adult female mosquitoes at 3.9- or 1.9-fold reduced concentrations on day 4 and at 2.1- or 2.4-fold reduced concentrations on day 10. Compared with the results for the WT, the median lethal time (LT 50 ) of Bb-AaIT was reduced by 28.6% at 1 × 10 7 conidia ml -1 and 34.3% at 1 × 10 6 conidia ml -1 in the larvae bioassay by conidia ingestion, while it decreased 32.3% at 1 × 10 7 conidia ml -1 by conidia ingestion and 24.2% at 1 × 10 8 conidia ml -1 by penetrating through the cuticle in the adult bioassay. All the differences were significant. Our findings indicated that Bb-AaIT had higher virulence and faster action than the WT in killing the larval and adult mosquitoes, and therefore, it is valuable for development as a commercial mosquito pesticide.

  17. Chrysodeixis chalcites nucleopolyhedrovirus (ChchNPV): Natural occurrence and efficacy as a biological insecticide on young banana plants in greenhouse and open-field conditions on the Canary Islands.

    Science.gov (United States)

    Fuentes, Ernesto Gabriel; Hernández-Suárez, Estrella; Simón, Oihane; Williams, Trevor; Caballero, Primitivo

    2017-01-01

    Chrysodeixis chalcites, an important pest of banana crops on the Canary Islands, is usually controlled by chemical insecticides. The present study aimed to evaluate the efficacy of the most prevalent isolate of the Chrysodeixis chalcites nucleopolyhedrovirus (ChchNPV, Baculoviridae) as a biological insecticide. Overall the prevalence of ChchNPV infection in C. chalcites populations was 2.3% (103 infected larvae out of 4,438 sampled), but varied from 0-4.8% on Tenerife and was usually low (0-2%) on the other islands. On Tenerife, infected larvae were present at 11 out of 17 plantations sampled. The prevalence of infection in larvae on bananas grown under greenhouse structures was significantly higher (3%) than in open-field sites (1.4%). The ChchNPV-TF1 isolate was the most abundant and widespread of four genetic variants of the virus. Application of 1.0x109 viral occlusion bodies (OBs)/l of ChchNPV-TF1 significantly reduced C. chalcites foliar damage in young banana plants as did commonly used pesticides, both in greenhouse and open-field sites. The insecticidal efficacy of ChchNPV-TF1 was similar to that of indoxacarb and a Bacillus thuringiensis (Bt)-based insecticide in one year of trials and similar to Bt in the following year of trails in greenhouse and field crops. However, larvae collected at different time intervals following virus treatments and reared in the laboratory experienced 2-7 fold more mortality than insects from conventional insecticide treatments. This suggests that the acquisition of lethal dose occurred over an extended period (up to 7 days) compared to a brief peak in larvae on plants treated with conventional insecticides. These results should prove useful for the registration of a ChchNPV-based insecticide for integrated management of this pest in banana crops on the Canary Islands.

  18. Chrysodeixis chalcites nucleopolyhedrovirus (ChchNPV: Natural occurrence and efficacy as a biological insecticide on young banana plants in greenhouse and open-field conditions on the Canary Islands.

    Directory of Open Access Journals (Sweden)

    Ernesto Gabriel Fuentes

    Full Text Available Chrysodeixis chalcites, an important pest of banana crops on the Canary Islands, is usually controlled by chemical insecticides. The present study aimed to evaluate the efficacy of the most prevalent isolate of the Chrysodeixis chalcites nucleopolyhedrovirus (ChchNPV, Baculoviridae as a biological insecticide. Overall the prevalence of ChchNPV infection in C. chalcites populations was 2.3% (103 infected larvae out of 4,438 sampled, but varied from 0-4.8% on Tenerife and was usually low (0-2% on the other islands. On Tenerife, infected larvae were present at 11 out of 17 plantations sampled. The prevalence of infection in larvae on bananas grown under greenhouse structures was significantly higher (3% than in open-field sites (1.4%. The ChchNPV-TF1 isolate was the most abundant and widespread of four genetic variants of the virus. Application of 1.0x109 viral occlusion bodies (OBs/l of ChchNPV-TF1 significantly reduced C. chalcites foliar damage in young banana plants as did commonly used pesticides, both in greenhouse and open-field sites. The insecticidal efficacy of ChchNPV-TF1 was similar to that of indoxacarb and a Bacillus thuringiensis (Bt-based insecticide in one year of trials and similar to Bt in the following year of trails in greenhouse and field crops. However, larvae collected at different time intervals following virus treatments and reared in the laboratory experienced 2-7 fold more mortality than insects from conventional insecticide treatments. This suggests that the acquisition of lethal dose occurred over an extended period (up to 7 days compared to a brief peak in larvae on plants treated with conventional insecticides. These results should prove useful for the registration of a ChchNPV-based insecticide for integrated management of this pest in banana crops on the Canary Islands.

  19. Vasoactive intestinal polypeptide (VIP) innervation of the human eyelid glands.

    Science.gov (United States)

    Seifert, P; Spitznas, M

    1999-06-01

    This study was conducted to obtain morphological proof of innervating nerve fibres in the glands of the human eyelid (accessory lacrimal glands of Wolfring, meibomian glands, goblet cells, glands of Zeis, glands of Moll, sweat glands, glands of lanugo hair follicles) and identification of the secretomotorically active neuropeptide vasoactive intestinal polypeptide (VIP) as a common transmitter. Epoxy-embedded ultrathin sections of tissue samples from human eyelids were studied using electron microscopy. Paraffin sections fixed in Bouin-Hollande solution were immunostained with rabbit antiserum against VIP. With the electron microscope we were able to identify nerves in the glandular stroma of all the glands examined with the exception of goblet cells. Intraepithelial single axons were only seen in the parenchyma of Wolfring glands. The morphological findings corresponded with the immunological finding of VIP-positive, nerve-like structures in the same locations, with the exception of lanugo hair follicle glands, and goblet cells. Our findings indicate that the glands of the eyelids and main lacrimal gland represent a functional unit with VIP as a possible common stimulating factor. Copyright 1999 Academic Press.

  20. Recovery of commercially produced Bacillus thuringiensis var. israelensis and Bacillus sphaericus from tires and prevalence of bacilli in artificial and natural containers.

    Science.gov (United States)

    Siegel, J P; Smith, A R; Novak, R J

    2001-03-01

    We conducted surveys to identify the species of spore-forming bacteria present in natural and artificial containers. Most of our samples came from Illinois. Identification was based on the cellular fatty acid composition of the bacterial cell wall. In addition, we utilized a custom database for commercially produced strains of Bacillus thuringiensis var. israelensis (Bti) and B. sphaericus, to differentiate between larvicidal isolates with commercial or native origin. Native Bti was present at low levels in almost all habitats but was not recovered from bromeliads and metal containers. In temporary woodland pools, 27.9% of the colonies recovered were native Bti. We did not recover larvicidal B. sphaericus in untreated habitats. VectoBac and VectoLex were applied to tires containing water and the tires were sampled 3 months and 9 months after treatment. Isolates of Bti and B. sphaericus with commercial origin were recovered as long as 9 months after application. We noticed numerous cadavers of Aedes triseriatus in several tires 9 months after treatment with VectoBac. We could not determine if this mortality resulted from recycling of Bti in these tires or whether insecticidal crystal proteins from the original treatment were resuspended. Bacillus thuringiensis var. israelensis isolates with commercial ancestry were recovered from untreated tires 9 months after application. Isolates of larvicidal B. sphaericus that differed from the bacteria in VectoLex were also recovered from untreated tires.

  1. Peptides PHI and VIP: comparison between vascular and nonvascular smooth muscle effect in rabbit uterus

    International Nuclear Information System (INIS)

    Bardrum, B.; Ottesen, B.; Fahrenkrug, J.

    1986-01-01

    The distribution and effects of the two neuropeptides, vasoactive intestinal polypeptide (VIP) and peptide histidine isoleucine amide (PHI), on vascular and nonvascular smooth muscle in the urogenital tract of nonpregnant rabbit female, were investigated. Immunoreactive VIP and PHI were present in all regions except the ovary with the highest concentration in the uterin cervix. By using in vitro tension recordings of myometrial specimens, it was demonstrated that both peptides displayed a dose-dependent inhibition of the mechanical activity. The dose-response curves of VIP and PHI were superimposable with and ID 50 of 3 x 10 -8 mol/l, and their combined effect was additive. In addition, the influence of the two peptides on myometrial blood flow (MBF) was investigated by the xenon-133 washout technique. Both peptides were found to increase MBF with the same potency and efficacy. Their combined effect was additive. In conclusion VIP and PHI are present in the rabbit urogenital tract, and the two peptides are equipotent inhibitors of mechanical nonvascular and vascular smooth muscle activity in the uterus

  2. Characterization of Bacillus thuringiensis strains from Jordan and ...

    African Journals Online (AJOL)

    Eight serotypes with Bacillus thuringiensis israelensis being the most common. Out of the twenty-six isolated strains, five strains (serotype: kenyae, kurstaki, kurstaki HD1 and thuringiensis) that produced bipyramid crystal proteins were toxic to the lepidoptera larvae of Ephestia kuehniella Zeller. The SDS-PAGE protein ...

  3. Association of bioassays and molecular characterization to select new Bacillus thuringiensis isolates effective against Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae); Associacao de bioensaios e caracterizacao molecular para selecao de novos isolados de Bacillus thuringiensis efetivos contra Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae)

    Energy Technology Data Exchange (ETDEWEB)

    Fatoretto, Julio C.; Sena, Janete A.D.; Lemos, Manoel V.F. [UNESP, Jaboticabal, SP (Brazil). Faculdade de Ciencias Agrarias e Veterinarias. Dept. de Biologia Aplicada a Agropecuaria; Barreto, Marliton R. [Universidade Federal do Mato Grosso (UFMT), Cuiaba, MT (Brazil). Inst. Universitario do Norte Matogrossense (IUNMAT)]. E-mail: mrbarreto@pop.com.br; Junior Boica, Arlindo L. (UNESP), Jaboticabal, SP (Brazil). Faculdade de Ciencias Agrarias e Veterinarias. Dept. de Fitossanidade)

    2007-09-15

    The fall armyworm, Spodoptera frugiperda (J. E. Smith), is one of the main corn pests and Bacillus thuringiensis is important in its control because of its entomopathogenic property. The objective of this study was the molecular characterization of B. thuringiensis isolates for cry1 locus presence and the assessment of the efficiency of these isolates in controlling S. frugiperda caterpillars. Gral-cry1 was used in the PCR analyses to confirm the presence of the cry1 locus in 15 isolates. A 3 x 108 spore/ml suspension bathed the diet used to feed 30 caterpillars per isolate, with three replications. The cry1 locus type genes of the different isolates were identified for five gene subclasses; linear regression analyses were carried out to ascertain possible associations between the presence of an individual cry1 locus gene and high levels of toxicity. All the DNAs amplified with Gral-cry1 presented an amplification product with the expected size. Regarding the levels of insecticide efficiency against the cob worm, 41 isolates presented 100% mortality and 16 presented an index between 70% and 90%. The cry1Ab gene was present in 80 isolates, cryb in 69 isolates, cry1Ac in all the isolates and cryv and cry1E in 93 and 27 isolates, respectively. The values regarding the individual effect of each gene on caterpillar mortality were significant at 1% probability for the cry1Ac and cry1E genes. (author)

  4. Possible Insecticidal Mechanisms Mediated by Immune-Response-Related Cry-Binding Proteins in the Midgut Juice of Plutella xylostella and Spodoptera exigua.

    Science.gov (United States)

    Lu, Keyu; Gu, Yuqing; Liu, Xiaoping; Lin, Yi; Yu, Xiao-Qiang

    2017-03-15

    Cry toxins are insecticidal toxin proteins produced by a spore-forming Gram-positive bacterium Bacillus thuringiensis. Interactions between the Cry toxins and the receptors from midgut brush border membrane vesicles (BBMVs), such as cadherin, alkaline phosphatase, and aminopeptidase, are key steps for the specificity and insecticidal activity of Cry proteins. However, little is known about the midgut juice proteins that may interfere with Cry binding to the receptors. To validate the hypothesis that there exist Cry-binding proteins that can interfere with the insecticidal process of Cry toxins, we applied Cry1Ab1-coupled Sepharose beads to isolate Cry-binding proteins form midgut juice of Plutella xylostella and Spodoptera exigua. Trypsin-like serine proteases and Dorsal were found to be Cry1Ab1-binding proteins in the midgut juice of P. xylostella. Peroxidase-C (POX-C) was found to be the Cry1Ab1-binding protein in the midgut juice of S. exigua. We proposed possible insecticidal mechanisms of Cry1Ab1 mediated by the two immune-related proteins: Dorsal and POX-C. Our results suggested that there exist, in the midgut juice, Cry-binding proteins, which are different from BBMV-specific receptors.

  5. Different toxicity of the novel Bacillus thuringiensis (Bacillales: Bacillaceae) strain LLP29 against Aedes albopictus and Culex quinquefasciatus (Diptera: Culicidae)

    Czech Academy of Sciences Publication Activity Database

    Zhang, L.; Tang, B.; Huang, E.; Huang, Z.; Liu, Z.; Huang, T.; Gelbič, Ivan; Guan, X.; Xu, L.

    2013-01-01

    Roč. 106, č. 3 (2013), s. 1098-1102 ISSN 0022-0493 Grant - others:National Natural Science Foundation of China(CN) 31071745; National Natural Science Foundation of China(CN) 31201574; Ministry of Education of China(CN) 20093515110010; Ministry of Education of China(CN) 20093515120010; Agricultural Science and Technology Achievements(CN) 2010GB2C400212; National High Technology Research and Development Program 863(CN) 2011AA10A203; Universities of Fujian Province(CN) JA12092; Fujian Agriculture and Forestry University(CN) xjq201203; Universities for the Development of the West Strait(CN) 0b08b005 Institutional research plan: CEZ:AV0Z50070508 Keywords : Bacillus thuringiensis * receptor binding * ELISA Subject RIV: ED - Physiology Impact factor: 1.605, year: 2013 http://www.bioone.org/doi/pdf/10.1603/EC12308

  6. Isolation and partial characterization of a mutant of Bacillus thuringiensis producing melanin Isolamento e caracterização parcial de um mutante de Bacillus thuringiensis produtor de melanina

    Directory of Open Access Journals (Sweden)

    Gislayne T. Vilas-Bôas

    2005-09-01

    Full Text Available A mutant (407-P of Bacillus thuringiensis subsp. thuringiensis strain 407 producing a melanin was obtained after treatment with the mutagenic agent ethyl-methane-sulfonate. Several microbiological and biochemical properties of the two strains were analyzed and the results were similar. The mutant 407-P was also incorporated into non-sterilized soil samples, recovered, easily identified, and quantified, what enables its use in ecology of B. thuringiensis.Um mutante (407-P da linhagem Bacillus thuringiensis subsp. thuringiensis 407 produtor de melanina foi obtido após tratamento com o agente mutagênico etil-metano-sulfonato. Diversas propriedades microbiológicas e bioquímicas das duas linhagens foram analisadas e os resultados foram similares. O mutante 407-P foi incorporado em amostras de solo não esterilizado, recuperado, facilmente identificado e quantificado, possibilitando seu uso em estudos de ecologia de B. thuringiensis.

  7. Clinical significance of changes of plasma CGRP and VIP levels in infants with bronchiolitis

    International Nuclear Information System (INIS)

    Yang Chun; Gu Ling; Zhang Yanjun; Xin Haiyan

    2005-01-01

    Objective: To explore the clinical significance of changes of calcitonin gene-related peptide (CGRP) and vasoactive intestinal peptide (VIP) levels in infants (2-24months) with bronchiolitis. Methods: Plasma levels of CGRP and VIP were determined with RIA in 31 infants with bronchiolitis both during acute infection and convalescence as well as in 35 controls. Results: Plasma CGRP levels in patients during acute infection were significantly higher than those in patients during convalescence and in controls (P<0.05). Levels of CGRP dropped during convalescence, but still remained significantly higher than those in controls (P<0.05). The reverse was true for the plasma VIP levels. The plasma VIP levels in patients during acute infection were significantly lower than those in patients during convalescence and in controls (P<0.05). During convalescence, the plasma VIP levels rose but remained significantly lower than those in controls (P<0.05). Conclusion: There were dynamic changes of plasma CGRP and VIP levels in the course of infant bronchiolitis and the two peptides played opposite roles. (authors)

  8. The diverse nematicidal properties and biocontrol efficacy of Bacillus thuringiensis Cry6A against the root-knot nematode Meloidogyne hapla.

    Science.gov (United States)

    Yu, Ziquan; Xiong, Jing; Zhou, Qiaoni; Luo, Haiyan; Hu, Shengbiao; Xia, Liqiu; Sun, Ming; Li, Lin; Yu, Ziniu

    2015-02-01

    Cry6A toxin from Bacillus thuringiensis is a representative nematicidal crystal protein with a variety of nematicidal properties to free-living nematode Caenorhabditis elegans. Cry6A shares very low homology and different structure with Cry5B, another representative nematicidal crystal protein, and probably acts in a distinct pathway. All these strongly indicate that Cry6A toxin is likely a potent candidate for nematicide. The present study dealt with global investigation to determine the detrimental impacts of Cry6Aa2 toxin on Meloidogyne hapla, a root-knot nematode, and evaluated its biocontrol efficacy in pot experiment. Obtained results indicated that Cry6Aa2 toxin exhibits obvious toxicity to second-stage juvenile of M. hapla, and significantly inhibits egg hatch, motility, and penetration to host plant. Pot experiment suggested that soil drenching with spore-crystal mixture of Cry6Aa2 can clearly lighten the disease of root-knot nematode, including reduction of galling index and egg masses on host plant root, decreasing final population of nematode in soil. Moreover, application of Cry6Aa2 can obviously promote plant growth. These results demonstrated that Cry6Aa2 toxin is a promising nematicidal agent, and possesses great potential in plant-parasitic nematode management and construction of transgenic crop with constant resistance to nematode. Copyright © 2015 Elsevier Inc. All rights reserved.

  9. Vacuum Insulation Panels (VIPs) for building construction industry - A review of the contemporary developments and future directions

    International Nuclear Information System (INIS)

    Alam, M.; Singh, H.; Limbachiya, M.C.

    2011-01-01

    Highlights: → Vacuum Insulation Panels (VIP), a high thermal resistance building insulation. → Review of research into VIPs for building applications. → High cost and uncertainty of service life are two barriers for VIP use in buildings. → SiO 2 /SiN x coated PET laminate- candidate for high barrier VIP envelope. → The optimum combination of VIP core and envelope yet to be determined. -- Abstract: Demand for energy efficient buildings has increased drastically in recent years and this trend will continue in the future. Insulating building elements will play a key role in meeting this demand by reducing heat losses through the building fabric. Due to their higher thermal resistance, Vacuum Insulation Panels (VIPs) would be a more energy efficient alternative to conventional building insulation materials. Thus, efforts to develop VIPs with characteristics suitable for applications to new and existing buildings are underway. This paper provides a review of important contemporary developments towards producing VIPs using various materials such as glass fibre, foams, perlite and fibre/powder composites. The limitations of the materials currently used to fabricate VIPs have not been emphasised in detail in previous review papers published. Selection criteria, methods to measure important properties of VIPs and analytical and numerical models presented in the past have been detailed. Limitations of currently employed design tools along with potential future materials such as Nano/microcellular foams and SiO x /SiN x coatings for use in VIPs are also described.

  10. Isolation, characterization and toxicity of native Bacillus thuringiensis isolates from different hosts and habitats in Iran

    Directory of Open Access Journals (Sweden)

    Ghassemi-Kahrizeh Akbar

    2017-09-01

    Full Text Available Bacillus thuringiensis is a Gram-positive, aerobic, facultative anaerobic and endospore-forming bacterium. Different strains of this species have the ability to produce parasporal crystalline inclusions which are toxic to larvae of different insect orders and other invertebrates and cause rapid death of the host. To determine the importance of this species in microbial control, we collected native strains and studied their virulence on the diamondback moth, Plutella xylostella. More than 148 samples were collected from Alborz, Guilan and Mazandaran Provinces. Experimental samples, including soil samples from forests, fruit gardens, agricultural fields, diseased and dead larvae, were transferred to a laboratory in sterile plastic containers. For evaluating B. thuringiensis isolates virulence, a cabbage leaf dip method with 106 cell · ml−1 concentration of various Bt isolates was applied to diamondback moths. Larval mortality was recorded 72 h after treatment. Based on bioassay results, all isolates were classified into three high, medium and low virulence groups. Protein level characterization based on the SDS-PAGE gel analysis showed that two isolates from a high virulence group have proteins of high molecular masses of 121 and 109 kDa. Results revealed that there is a positive correlation between protein masses and virulence of isolates. In addition, this research introduced nine strains that are highly toxic to P. xylostella and would be valuable as insecticidal agents for controlling lepidopteran pests.

  11. A parasporin from Bacillus thuringiensis native to Peninsular India ...

    Indian Academy of Sciences (India)

    Thomas Chubicka

    2018-05-03

    May 3, 2018 ... Apoptosis; Bacillus thuringiensis; crystal protein; cytotoxicity; ... It acts by creating pores in the intestinal duct ... however diverse types of mechanisms of action have been ... parasporins that can be utilized in the cancer drug.

  12. Association of bioassays and molecular characterization to select new Bacillus thuringiensis isolates effective against Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae)

    International Nuclear Information System (INIS)

    Fatoretto, Julio C.; Sena, Janete A.D.; Lemos, Manoel V.F.; Junior Boica, Arlindo L. , Jaboticabal, SP . Faculdade de Ciencias Agrarias e Veterinarias. Dept. de Fitossanidade; Brazil)

    2007-01-01

    The fall armyworm, Spodoptera frugiperda (J. E. Smith), is one of the main corn pests and Bacillus thuringiensis is important in its control because of its entomopathogenic property. The objective of this study was the molecular characterization of B. thuringiensis isolates for cry1 locus presence and the assessment of the efficiency of these isolates in controlling S. frugiperda caterpillars. Gral-cry1 was used in the PCR analyses to confirm the presence of the cry1 locus in 15 isolates. A 3 x 108 spore/ml suspension bathed the diet used to feed 30 caterpillars per isolate, with three replications. The cry1 locus type genes of the different isolates were identified for five gene subclasses; linear regression analyses were carried out to ascertain possible associations between the presence of an individual cry1 locus gene and high levels of toxicity. All the DNAs amplified with Gral-cry1 presented an amplification product with the expected size. Regarding the levels of insecticide efficiency against the cob worm, 41 isolates presented 100% mortality and 16 presented an index between 70% and 90%. The cry1Ab gene was present in 80 isolates, cryb in 69 isolates, cry1Ac in all the isolates and cryv and cry1E in 93 and 27 isolates, respectively. The values regarding the individual effect of each gene on caterpillar mortality were significant at 1% probability for the cry1Ac and cry1E genes. (author)

  13. Sensory experience regulates cortical inhibition by inducing IGF1 in VIP neurons.

    Science.gov (United States)

    Mardinly, A R; Spiegel, I; Patrizi, A; Centofante, E; Bazinet, J E; Tzeng, C P; Mandel-Brehm, C; Harmin, D A; Adesnik, H; Fagiolini, M; Greenberg, M E

    2016-03-17

    Inhibitory neurons regulate the adaptation of neural circuits to sensory experience, but the molecular mechanisms by which experience controls the connectivity between different types of inhibitory neuron to regulate cortical plasticity are largely unknown. Here we show that exposure of dark-housed mice to light induces a gene program in cortical vasoactive intestinal peptide (VIP)-expressing neurons that is markedly distinct from that induced in excitatory neurons and other subtypes of inhibitory neuron. We identify Igf1 as one of several activity-regulated genes that are specific to VIP neurons, and demonstrate that IGF1 functions cell-autonomously in VIP neurons to increase inhibitory synaptic input onto these neurons. Our findings further suggest that in cortical VIP neurons, experience-dependent gene transcription regulates visual acuity by activating the expression of IGF1, thus promoting the inhibition of disinhibitory neurons and affecting inhibition onto cortical pyramidal neurons.

  14. Increase of prolactin mRNA in the rat hypothalamus after intracerebroventricular injection of VIP or PACAP.

    Science.gov (United States)

    Bredow, S; Kacsóh, B; Obál, F; Fang, J; Krueger, J M

    1994-10-17

    Vasoactive intestinal peptide (VIP), the structurally homologous pituitary adenylate cyclase-activating peptide (PACAP) and the pituitary hormone, prolactin (PRL) enhance rapid eye movement sleep (REMS). VIP and PACAP are both inducers of PRL gene expression and release in the pituitary gland. Little is known about PRL regulation in the brain although it is hypothesized that the REMS-promoting activity of i.c.v. administered VIP may be mediated via the activation of cerebral PRL. To test whether VIP or PACAP in fact increase intracerebral mRNA, the peptides (VIP: 30 or 300 pmol; PACAP: 220 pmol) were injected i.c.v. into rats at dark onset. 1 h later, cDNA was synthesized from purified hypothalamic mRNA. Standardized amounts were analysed for PRL using the polymerase chain reaction followed by Southern blotting and hybridization. Compared with beta-actin mRNA levels, both VIP and PACAP increased PRL mRNA levels in a dose-dependent fashion though VIP was more effective on a molar basis. The previously reported alternatively spliced PRL mRNA (lacking exon 4) was not detected. The data support the hypothesis that the REMS-promoting activity of central VIP and PACAP might be mediated by cerebral PRL.

  15. A differentially displayed mRNA related to restistance to Bacillus thuringiensis israelensis of Aedes albopictus selected in vitro-activated CYT1AA6

    Czech Academy of Sciences Publication Activity Database

    Zhang, L.; Zhang, Q.; Huang, E.; Li, M.; Huang, T.; Xu, L.; Wu, Ch.; Guan, X.; Gelbič, Ivan

    2012-01-01

    Roč. 28, č. 4 (2012), s. 327-329 ISSN 8756-971X Institutional support: RVO:60077344 Keywords : Bacillus thuringiensis israelensis * resistence * Aedes albopictus Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 0.755, year: 2012 http://www.bioone.org/doi/pdf/10.2987/12-6263R.1

  16. Synthesis, herbicidal, fungicidal and insecticidal evaluation of 3-(dichlorophenyl)- isocoumarins and (±)-3-(dichlorophenyl)-3,4-dihydroisocoumarins

    International Nuclear Information System (INIS)

    Qadeer, Ghulam; Rama, Nasim Hasan; Fan, Zhi-Jin; Liu, Bin; Liu, Xiu-Feng

    2007-01-01

    This is the first report showing that 3-(dichlorophenyl)isocoumarins and (±)-3,4-dihydroisocoumarins are plant and plant fungus growth inhibitors. 3-Dichlorophenylisocoumarins were synthesized by condensation of homophthalic acid with dichlorobenzoyl chlorides. The alkaline hydrolysis of these isocoumarins afforded keto acids. Racemic 3-(Dichlorophenyl)-3,4-dihydroisocoumarins were obtained by reduction of keto acids to racemic hydroxy acids, followed by cyclodehydration using acetic anhydride. The herbicidal, fungicidal and insecticidal activities of the synthesized compounds have been evaluated. Some of the synthesized compounds show excellent herbicidal and fungicidal activities but none of the synthesized compounds presented any insecticidal effects on the test insects. The findings of this study suggest that isocoumarins and related compounds may serve as lead compounds towards the design of bioactive herbicides and fungicides. (author)

  17. Concentration-Response and Residual Activity of Insecticides to Control Herpetogramma phaeopteralis (Lepidoptera: Crambidae) in St. Augustinegrass.

    Science.gov (United States)

    Tofangsazi, Nastaran; Cherry, Ron H; Beeson, Richard C; Arthurs, Steven P

    2015-04-01

    Tropical sod webworm, Herpetogramma phaeopteralis Guenée, is an important pest of warm-season turfgrass in the Gulf Coast states of the United States, the Caribbean Islands, and Central America. Current control recommendations rely on topical application of insecticides against caterpillars. The objective of this study was to generate resistance baseline data of H. phaeopteralis to six insecticide classes. Residual activity of clothianidin, chlorantraniliprole, and bifenthrin was also compared under field conditions in Central Florida. Chlorantraniliprole was the most toxic compound tested (LC50 value of 4.5 ppm), followed by acephate (8.6 ppm), spinosad (31.1 ppm), clothianidin (46.6 ppm), bifenthrin (283 ppm) and Bacillus thuringiensis kurstaki, (342 ppm). In field tests, all compounds at label rates were effective (≥94% mortality of larvae exposed to fresh residues). However, a more rapid decline in activity of clothianidin and bifenthrin was observed compared with chlorantraniliprole. Clothianidin had no statistically detectable activity after 4 wk post-application in spring and the fall, and bifenthrin had no detectable activity after 3 wk in the spring and the fall. However, chlorantraniliprole maintained significant activity (≥84% mortality) compared with other treatments throughout the 5-wk study period. This study provides new information regarding the relative toxicities and persistence of current insecticides used for H. phaeopteralis and other turfgrass caterpillars. © 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.

  18. Construction of a recombinant Bacillus velezensis strain as an integrated control agent against plant diseases and insect pests.

    Science.gov (United States)

    Roh, Jong Yul; Liu, Qin; Choi, Jae Young; Wang, Yong; Shim, Hee Jin; Xu, Hong Guang; Choi, Gyung Ja; Kim, Jin-Cheol; Je, Yeon Ho

    2009-10-01

    To construct a new recombinant strain of Bacillus velezensis that has antifungal and insecticidal activity via the expression of the insecticidal Bacillus thuringiensis crystal protein, a B. thuringiensis expression vector (pHT1K-1Ac) was generated that contained the B. thuringiensis cry1Ac gene under the control of its endogenous promoter in a minimal E. coli-B. thuringiensis shuttle vector (pHT1K). This vector was introduced into a B. velezensis isolate that showed high antifungal activities against several plant diseases, including rice blast (Magnaporthe grisea), rice sheath blight (Rhizotonia solani), tomato gray mold (Botrytis cinerea), tomato late blight (Phytophthora infestans), and wheat leaf rust (Puccinia recondita), by electroporation. The recombinant B. velezensis strain was confirmed by PCR using cry1Ac-specific primers. Additionally, the recombinant strain produced a protein approximately 130 kDa in size and parasporal inclusion bodies similar to B. thuringiensis. The in vivo antifungal activity assay demonstrated that the activity of the recombinant B. velezensis strain was maintained at the same level as that of wild-type B. velezensis. Furthermore, it exhibited high insecticidal activity against a lepidopteran pest, Plutella xylostella, although its activity was lower than that of a recombinant B. thuringiensis strain, whereas wild-type B. velezensis strain did not show any insecticidal activity. These results suggest that this recombinant B. velezensis strain can be used to control harmful insect pests and fungal diseases simultaneously in one crop.

  19. Searches for the Violation of Pauli Exclusion Principle at LNGS in VIP(-2) experiment

    CERN Document Server

    Shi, H; Bertolucci, S; Berucci, C; Bragadireanu, A M; Cargnelli, M; Clozza, A; Curceanu, C; De Paolis, L; Di Matteo, S; d'Uffizi, A; Egger, J P; Guaraldo, C; Iliescu, M; Ishiwatari, T; Marton, J; Laubenstein, M; Milotti, E; Pietreanu, D; Piscicchia, K; Ponta, T; Vidal, A.Romero; Sbardella, E; Scordo, A; Sirghi, D L; Sirghi, F; Sperandio, L; Vazquez Doce, O; Widmann, E; Zmeskal, J

    2016-01-01

    The VIP (Violation of Pauli exclusion principle) experiment and its follow-up experiment VIP-2 at the Laboratori Nazionali del Gran Sasso (LNGS) search for X-rays from Cu atomic states that are prohibited by the Pauli Exclusion Principle (PEP). The candidate events, if they exist, will originate from the transition of a $2p$ orbit electron to the ground state which is already occupied by two electrons. The present limit on the probability for PEP violation for electron is 4.7 $\\times10^{-29}$ set by the VIP experiment. With upgraded detectors for high precision X-ray spectroscopy, the VIP-2 experiment will improve the sensitivity by two orders of magnitude.

  20. VIP visit of LHC Computing Grid Project

    CERN Multimedia

    Krajewski, Yann Tadeusz

    2015-01-01

    VIP visit of LHC Computing Grid Project with Dr -.Ing. Tarek Kamel [Senior Advisor to the President for Government Engagement, ICANN Geneva Office] and Dr Nigel Hickson [VP, IGO Engagement, ICANN Geneva Office

  1. Selection of Bacillus thuringiensis strains toxic to cotton boll weevil (Anthonomus grandis, Coleoptera: Curculionidae) larvae.

    Science.gov (United States)

    Pérez, Melisa P; Sauka, Diego H; Onco, María I; Berretta, Marcelo F; Benintende, Graciela B

    Preliminary bioassays with whole cultures (WC) of 124 Bacillus thuringiensis strains were performed with neonate larvae of Anthonomus grandis, a major cotton pest in Argentina and other regions of the Americas. Three exotic and four native strains were selected for causing more than 50% mortality. All of them were β-exotoxin producers. The native strains shared similar morphology of parasporal crystals, similar protein pattern and identical insecticidal gene profiles. These features resembled Lepidoptera-toxic strains. Furthermore, these strains showed a Rep-PCR pattern identical to lepidoptericidal strain HD-1, suggesting that these strains may belong to serovar kurstaki. However, some differences were observed in the plasmid profiles and in the production of β-exotoxin. To determine the culture fractions where the insecticidal metabolites were present, bioassays including resuspended spore-crystal pellets, filtered supernatants (FS) were compared with those of WC. Both fractions tested showed some level of insecticidal activity. The results may suggest that the main toxic factors can be found in FS and could be directly correlated with the presence of β-exotoxin. Based on the bioassays with FS and autoclaved FS, the participation of thermolabile virulence factors such as Cry1I in toxicity is neither discarded. In the selected strains, β-exotoxin would be the major associated virulence factor; therefore, their use in biological control of A. grandis should be restricted. Nevertheless, these strains could be the source of genes (e.g., cry1Ia) to produce transgenic cotton plants resistant to this pest. Copyright © 2017 Asociación Argentina de Microbiología. Publicado por Elsevier España, S.L.U. All rights reserved.

  2. Vasoactive intestinal peptide (VIP) labelling with iodine-131 by direct method

    International Nuclear Information System (INIS)

    Colturato, M.T.; Silva, C.P.G. da; Araujo, E.B.

    2002-01-01

    The Vasoactive Intestinal Peptide (VIP) is a 28-amino acid polypeptide with a great numbers of receptors in tumoral cells, including adenocarcinomas and pancreatic and colon carcinomas. The VIP molecule contains two tyrosine residues, in positions 10 and 22, that are theoretically equally susceptible to iodination, The VIP was labeled with 131-iodine by direct method using Iodogen as oxidant agent: 15.03 mmol VIP + 0.10 nmol KI + [ 131 I]NaI + 13.9 mmol Iodogen; the final volume was adjust to 100 μL using 0.2 M phosphate buffer, pH 7.5 and the reaction proceed with stirring for 30 minutes at room temperature. The radiochemical purity was determined by electrophoresis (Whatman 1MM paper; 0.05 M barbital buffer; pH 8.6; 150 V; 40 minutes) that indicates low percent of free 131-iodine. The high performance liquid chromatography (HPLC) system using RPC 18 , 10 μm, 4 x 250mm column, was able to separate the different radiochemical species, only when an isocratic mixture of acetonitrile: 0.1% trifluoroacetic acid (27:73) was used, with 0.5 mL/min. flux. (author)

  3. MENTOR-VIP: Piloting a global mentoring program for injury and violence prevention.

    Science.gov (United States)

    Hyder, Adnan A; Meddings, David; Bachani, Abdulgafoor M

    2009-06-01

    Injuries occur as the result of a confluence of factors: environmental, social, biological, economic, and behavioral. To effectively address the burden of injuries, especially in low- and middle-income countries, a focus is needed on developing the human resource capacity for injury prevention. MENTOR-VIP is a global mentoring program that the authors developed with this need in mind. MENTOR-VIP approaches developing human resources in injury prevention by providing mentoring opportunities for junior professionals involved in its practice, research, and/or programs. MENTOR-VIP entails a 12-month working relationship between junior injury prevention practitioners (mentees) and more experienced individuals in the field (mentors). Its general objective is to improve global human resource capacity to effectively prevent and control injury and violence through the enhanced development of relevant skills. The program is currently in its pilot phase and is nearing the end of its second formal mentoring cycle, which began on September 1, 2008. This article discusses mentoring professionals as a key strategy to developing the human resource component of capacity, and one which complements existing approaches to capacity development. The authors also provide an overview of the rationale, modalities, objectives, and evaluation of MENTOR-VIP. This article highlights the importance of capacity building in the injury prevention field and situates MENTOR-VIP within the larger context of capacity building for global public health.

  4. Intracerebroventricular Infusion of Vasoactive Intestinal Peptide (VIP Rescues the Luteinizing Hormone Surge in Middle-Aged Female Rats

    Directory of Open Access Journals (Sweden)

    Yan eSun

    2012-02-01

    Full Text Available Reproductive aging is characterized by delayed and attenuated luteinizing hormone (LH surges apparent in middle-aged rats. The suprachiasmatic nucleus (SCN contains the circadian clock that is responsible for the timing of diverse neuroendocrine rhythms. Electrophysiological studies suggest vasoactive intestinal peptide (VIP originating from the SCN excites gonadotropin-releasing hormone (GnRH neurons and affects daily patterns of GnRH-LH release. Age-related LH surge dysfunction correlates with reduced VIP mRNA expression in the SCN and fewer GnRH neurons with VIP contacts expressing c-fos, a marker of neuronal activation, on the day of the LH surge. To determine if age-related LH surge dysfunction reflects reduced VIP availability or altered VIP responsiveness under estradiol positive feedback conditions, we assessed the effect of intracerebroventricular (icv VIP infusion on c-fos expression in GnRH neurons and on LH release in ovariohysterectomized, hormone-primed young and middle-aged rats. Icv infusion of VIP between 1300 and 1600 h significantly advanced the time of peak LH release, increased total and peak LH release, and increased the number of GnRH neurons expressing c-fos on the day of the LH surge in middle-aged rats. Surprisingly, icv infusion of VIP in young females significantly reduced the number of GnRH neurons expressing c-fos and delayed and reduced the LH surge. These observations suggest that a critical balance of VIP signaling is required to activate GnRH neurons for an appropriately timed and robust LH surge in young and middle-aged females. Age-related LH surge changes may, in part, result from decreased availability and reduced VIP-mediated neurotransmission under estradiol positive feedback conditions.

  5. 40 CFR 174.518 - Bacillus thuringiensis Cry3Bb1 protein in corn; exemption from the requirement of a tolerance.

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 23 2010-07-01 2010-07-01 false Bacillus thuringiensis Cry3Bb1 protein in corn; exemption from the requirement of a tolerance. 174.518 Section 174.518 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) PESTICIDE PROGRAMS PROCEDURES AND REQUIREMENTS FOR PLANT-INCORPORATED PROTECTANTS Tolerances and...

  6. Performance and cross-crop resistance of Cry1F-maize selected Spodoptera frugiperda on transgenic Bt cotton: implications for resistance management.

    Science.gov (United States)

    Yang, Fei; Kerns, David L; Brown, Sebe; Kurtz, Ryan; Dennehy, Tim; Braxton, Bo; Head, Graham; Huang, Fangneng

    2016-06-15

    Transgenic crops producing Bacillus thuringiensis (Bt) proteins have become a primary tool in pest management. Due to the intensive use of Bt crops, resistance of the fall armyworm, Spodoptera frugiperda, to Cry1F maize has occurred in Puerto Rico, Brazil, and some areas of the southeastern U.S. The sustainability of Bt crops faces a great challenge because the Cry1F-maize resistant S. frugiperda may also infest other Bt crops in multiple cropping ecosystems. Here we examined the survival and plant injury of a S. frugiperda population selected with Cry1F maize on three single-gene and five pyramided Bt cotton products. Larvae of Cry1F-susceptible (SS), -heterozygous (RS), and -resistant (RR) genotypes of S. frugiperda were all susceptible to the pyramided cotton containing Cry1Ac/Cry2Ab, Cry1Ac/Cry1F/Vip3A, Cry1Ab/Cry2Ae, or Cry1Ab/Cry2Ae/Vip3A, and the single-gene Cry2Ae cotton. Pyramided cotton containing Cry1Ac/Cry1F was effective against SS and RS, but not for RR. These findings show that the Cry1F-maize selected S. frugiperda can cause cross-crop resistance to other Bt crops expressing similar insecticidal proteins. Resistance management and pest management programs that utilize diversify mortality factors must be implemented to ensure the sustainability of Bt crops. This is especially important in areas where resistance to single-gene Bt crops is already widespread.

  7. Long-term estradiol treatment improves VIP-mediated vasodilation in atherosclerotic proximal coronary arteries

    DEFF Research Database (Denmark)

    Dalsgaard, T.; Mortensen, Alicja; Larsen, C. R.

    2003-01-01

    arteries. Female ovariectomized homozygous Watanabe heritable hyperlipidemic rabbits were randomized to 16 weeks treatment with 17beta-estradiol or placebo. The diet was semisynthetic, thereby avoiding the influence of phytoestrogens. Artery ring segments were mounted for isometric tension recordings...... in myographs. Following precontraction, the dose-response relationships for VIP and PACAP were evaluated. Treatment with 17beta-estradiol significantly improved the maximum VIP-mediated vasodilation (E-max, percentage of precontraction) in proximal coronary arteries (45.8 +/- 9.6% vs. 24.1 +/- 3.7%, p ....05). In the same artery segment, 17β-estradiol induced a significant decrease in the relative ratio between the repeated contractile response to potassium 30 and 120 mM (100 +/- 7% vs. 132 +/- 11%, p

  8. Estimation of insecticide persistence, biological activity and mosquito resistance to PermaNet® 2 long-lasting insecticidal nets over three to 32 months of use in Ethiopia.

    Science.gov (United States)

    Anshebo, Gedeon Yohannes; Graves, Patricia M; Smith, Stephen C; Wills, Aprielle B; Damte, Mesele; Endeshaw, Tekola; Shargie, Estifanos Biru; Gebre, Teshome; Mosher, Aryc W; Patterson, Amy E; Emerson, Paul M

    2014-03-06

    Information is needed on the expected durability of insecticidal nets under operational conditions. The persistence of insecticidal efficacy is important to estimate the median serviceable life of nets under field conditions and to plan for net replacement. Deltamethrin residue levels were evaluated by the proxy method of X-ray fluorescence spectrometry on 189 nets used for three to six months from nine sites, 220 nets used for 14-20 months from 11 sites, and 200 nets used for 26-32 months from ten sites in Ethiopia. A random sample of 16.5-20% of nets from each time period (total 112 of 609 nets) were tested by bioassay with susceptible mosquitoes, and nets used for 14-20 months and 26-32 months were also tested with wild caught mosquitoes. Mean insecticide levels estimated by X-ray fluorescence declined by 25.9% from baseline of 66.2 (SD 14.6) mg/m2 at three to six months to 44.1 (SD 21.2) mg/m2 at 14-20 months and by 30.8% to 41.1 (SD 18.9) mg/m2 at 26-32 months. More than 95% of nets retained greater than 10 mg/m2 of deltamethrin and over 79% had at least 25 mg/m2 at all time periods. By bioassay with susceptible Anopheles, mortality averaged 89.0% on 28 nets tested at three to six months, 93.3% on 44 nets at 14-20 months and 94.1% on 40 nets at 26-32 months. With wild caught mosquitoes, mortality averaged 85.4% (range 79.1 to 91.7%) at 14-20 months but had dropped significantly to 47.2% (39.8 to 54.7%) at 26-32 months. Insecticide residue level, as estimated by X-ray fluorescence, declined by about one third between three and six months and 14-20 months, but remained relatively stable and above minimum requirements thereafter up to 26-32 months. The insecticidal activity of PermaNet® 2.0 long-lasting insecticidal nets in the specified study area may be considered effective to susceptible mosquitoes at least for the duration indicated in this study (32 months). However, results indicated that resistance in the wild population is already rendering nets with

  9. Insecticide-Mediated Up-Regulation of Cytochrome P450 Genes in the Red Flour Beetle (Tribolium castaneum

    Directory of Open Access Journals (Sweden)

    Xiao Liang

    2015-01-01

    Full Text Available Some cytochrome P450 (CYP genes are known for their rapid up-regulation in response to insecticide exposures in insects. To date, however, limited information is available with respect to the relationships among the insecticide type, insecticide concentration, exposure duration and the up-regulated CYP genes. In this study, we examined the transcriptional response of eight selected CYP genes, including CYP4G7, CYP4Q4, CYP4BR3, CYP12H1, CYP6BK11, CYP9D4, CYP9Z5 and CYP345A1, to each of four insecticides in the red flour beetle, Tribolium castaneum. Reverse transcription quantitative PCR (RT-qPCR revealed that CYP4G7 and CYP345A1 can be significantly up-regulated by cypermethrin (1.97- and 2.06-fold, respectively, permethrin (2.00- and 2.03-fold and lambda-cyhalothrin (1.73- and 1.81-fold, whereas CYP4BR3 and CYP345A1 can be significantly up-regulated by imidacloprid (1.99- and 1.83-fold when 20-day larvae were exposed to each of these insecticides at the concentration of LC20 for 24 h. Our studies also showed that similar levels of up-regulation can be achieved for CYP4G7, CYP4BR3 and CYP345A1 by cypermethrin, permethrin, lambda-cyhalothrin or imidacloprid with approximately one fourth of LC20 in 6 h. Our study demonstrated that up-regulation of these CYP genes was rapid and only required low concentrations of insecticides, and the up-regulation not only depended on the CYP genes but also the type of insecticides. Our results along with those from previous studies also indicated that there were no specific patterns for predicting the up-regulation of specific CYP gene families based on the insecticide classification.

  10. Avaliação da suscetibilidade a inseticidas de populações da traça-das-crucíferas de algumas áreas do Brasil Forecasting insecticide susceptibility in Diamondback Moth populations from different areas of Brazil

    Directory of Open Access Journals (Sweden)

    Marina Castelo Branco

    2003-09-01

    Full Text Available A traça-das-crucíferas (TDC é a praga mais importante do repolho, sendo basicamente controlada por inseticidas. Em alguns casos, as lavouras são pulverizadas duas a quatro vezes por semana, sem sucesso. Isto acontece porque, em muitos casos, são empregados inseticidas ineficientes, para os quais a praga já apresenta resistência. Trabalhos anteriores demonstraram ser possível determinar os inseticidas ineficazes para o controle da TDC em testes de laboratório, através do uso da dosagem recomendada. Neste trabalho foram coletadas larvas e pupas do inseto nos estados do Ceará (Tianguá, Minas Gerais (Barroso, Bahia (Mucugê, Mato Grosso (Sinop e no Distrito Federal (Brazlândia e Embrapa Hortaliças. As populações foram criadas em laboratório e, dependendo do número de larvas de primeira geração, estas foram tratadas com as dosagens recomendadas de abamectin, acefato, B. thuringiensis, cartap, clorfluazuron, deltametrina e spinosad. Foi previamente assumido que um inseticida eficiente seria aquele que causasse a mortalidade de mais de 90% das larvas. Os resultados mostraram que a eficiência dos inseticidas variou entre as diferentes áreas. Spinosad causou a mortalidade de l00% das larvas em todos os locais. Foram ineficientes acefato, B. thuringiensis e cartap em Tianguá; abamectin em Brazlândia e clorfluazuron em Mucugê. Deltametrina não foi eficiente no controle das populações da praga coletadas em nenhuma das áreas geográficas amostradas. Conclui-se que as populações de traça-das-crucíferas brasileiras são resistentes a um ou mais ingredientes ativos e programas de manejo de resistência a inseticidas em TDC devem ser implementados.The Diamondback Moth (DBM is an important brassica pest and is controlled by several types of insecticides. On some occasions, the fields are sprayed two-four times per week without success. Sometimes this occurs because ineffective insecticides are used. Works showed that laboratory

  11. Improving the frequency of visual infusion phlebitis (VIP) scoring on an oncology ward

    Science.gov (United States)

    Tzolos, Evangelos; Salawu, Abdulazeez

    2014-01-01

    Phlebitis from peripheral intravenous infusions is an important potential source of oncology patient morbidity. Important factors found to determine phlebitis incidence include the kind of infusion and dwell time of intravenous cannula. Early studies showed incidence rates of between 25–70% worldwide, and association with up to 10% of S. aureus bacteraemia. The introduction of the visual infusion phlebitis (VIP) score tool for assessment of the early signs of phlebitis, along with prompt removal of peripheral intravenous cannulas, has been very successful in reducing the incidence below the acceptable rate of 5%. However, achieving this goal depends on strict compliance with guidelines for cannula insertion, documentation, and assessment using the VIP tool. This study aimed to increase the use of VIP scoring tool to 100% on an oncology ward during a four to six month period in order to maximise its utility in phlebitis prevention. Three plan-do-study-act (PDSA) cycles were carried out, during which two major interventions were introduced. The first cycle aimed to improve junior doctors’ awareness of VIP scoring using presentations in induction meetings and posters. The second cycle ensured that ready access to the VIP tool was provided in the form of bedside intentional rounding charts. Proportions of intravenous cannulas with proper documentation and VIP assessment were measured before intervention and at nine subsequent bi-weekly time points. Pre-intervention, under 30% of cannulas were properly documented and assessed. This proportion rose to around 80% by the end of the second PDSA cycle and achieved 100% by the end of the third cycle. PMID:26734282

  12. Comparison of Performance of the VIP and WMT in a Criminal Forensic Sample.

    Science.gov (United States)

    Fazio, Rachel L; Denney, Robert L

    2018-01-24

    To compare the efficacy of the Validity Indicator Profile (VIP) and Word Memory Test (WMT) in a criminal forensic population. Potential participants included 225 male evaluees from a maximum-security Federal prison referred for neuropsychological evaluation for either forensic purposes or for suspected neurocognitive dysfunction as part of a medical evaluation. Examinees were included in the analysis if administered the VIP (Verbal, Nonverbal, or both tests) and WMT along with at least two other freestanding PVTs; 74 satisfied these criteria. Participants were then categorized as having probable Malingered Neurocognitive Dysfunction (+MND) if they failed at least two freestanding validity indicators, negative for MND (-MND) if they passed all indicators, and Possible MND (pMND) if they failed one indicator. Groups were very demographically similar. There were significant differences in WMT scores and distribution of VIP profiles across groups. Whether using traditional or investigative cut scores, and whether using the WMT with or without consideration of a GMIP profile, the WMT demonstrated superior sensitivity and specificity on nearly every comparison. The VIP, when interpreted in the traditional fashion, and the WMT with GMIP, both had more than adequate psychometric properties when used with criminal forensic evaluees, strengthening the body of literature supporting their use for these types of evaluations. Counting a positive on either of the VIP subtests as an indication of +MND improves the psychometric properties of the VIP slightly, although the WMT had the better overall classification accuracy. © The Author(s) 2018. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  13. VIP: A knowledge-based design aid for the engineering of space systems

    Science.gov (United States)

    Lewis, Steven M.; Bellman, Kirstie L.

    1990-01-01

    The Vehicles Implementation Project (VIP), a knowledge-based design aid for the engineering of space systems is described. VIP combines qualitative knowledge in the form of rules, quantitative knowledge in the form of equations, and other mathematical modeling tools. The system allows users rapidly to develop and experiment with models of spacecraft system designs. As information becomes available to the system, appropriate equations are solved symbolically and the results are displayed. Users may browse through the system, observing dependencies and the effects of altering specific parameters. The system can also suggest approaches to the derivation of specific parameter values. In addition to providing a tool for the development of specific designs, VIP aims at increasing the user's understanding of the design process. Users may rapidly examine the sensitivity of a given parameter to others in the system and perform tradeoffs or optimizations of specific parameters. A second major goal of VIP is to integrate the existing corporate knowledge base of models and rules into a central, symbolic form.

  14. Development of Monoclonal Antibodies Recognizing Linear Epitope: Illustration by Three Bacillus thuringiensis Crystal Proteins of Genetically Modified Cotton, Maize, and Tobacco.

    Science.gov (United States)

    Cao, Zhen; Zhang, Wei; Ning, Xiangxue; Wang, Baomin; Liu, Yunjun; Li, Qing X

    2017-11-22

    Bacillus thuringiensis Cry1Ac, Cry1Ia1, and Cry1Ie are δ-endotoxin insecticidal proteins widely implemented in genetically modified organisms (GMO), such as cotton, maize, and potato. Western blot assay integrates electrophoresis separation power and antibody high specificity for monitoring specific exogenous proteins expressed in GMO. Procedures for evoking monoclonal antibody (mAb) for Western blot were poorly documented. In the present study, Cry1Ac partially denatured at 100 °C for 5 min was used as an immunogen to develop mAbs selectively recognizing a linear epitope of Cry1Ac for Western blot. mAb 5E9C6 and 3E6E2 selected with sandwich ELISA strongly recognized the heat semidenatured Cry1Ac. Particularly, 3E6E2 recognized both E. coli and cotton seed expressed Cry1Ac in Western blot. Such strategy of using partially denatured proteins as immunogens and using sandwich ELISA for mAb screening was also successfully demonstrated with production of mAbs against Cry1Ie for Western blot assay in maize.

  15. The Volunteering-in-Place (VIP) Program: Providing meaningful volunteer activity to residents in assisted living with mild cognitive impairment.

    Science.gov (United States)

    Klinedinst, N Jennifer; Resnick, Barbara

    2016-01-01

    The Volunteering-in-Place (VIP) Program was developed to provide individualized meaningful volunteer activities matched to interests and capabilities for older adults with MCI in assisted living. The purposes of this single-site pre-test/post-test pilot study were to (1) establish feasibility of the VIP Program based on treatment fidelity (design, treatment, delivery, enactment); and (2) evaluate preliminary efficacy via improvement in psychological health (depressive symptoms, usefulness, purpose, resilience, and life satisfaction) and decreased sedentary activity (survey and Fitbit) at 3 and 6 months. Ten residents participated. The majority was white, female and educated, and on average 88 years old. The VIP Program was feasible and most participants continued to volunteer at 6 months. There were non-significant improvements in depressive symptoms, usefulness, purpose, resilience and recreational physical activity. The results of this study provide support for the feasibility of the VIP Program. Further study is necessary to examine efficacy. Copyright © 2016 Elsevier Inc. All rights reserved.

  16. Screening of Bacillus thuringiensis strains effective against mosquitoes Prospecção de estirpes de Bacillus thuringiensis efetivas contra mosquitos

    Directory of Open Access Journals (Sweden)

    Rose Gomes Monnerat

    2005-02-01

    Full Text Available The objective of this work was to evaluate 210 Bacillus thuringiensis strains against Aedes aegypti and Culex quinquefasciatus larvae to select the most effective. These strains were isolated from different regions of Brazil and are stored in a Bacillus spp. collection at Embrapa Recursos Genéticos e Biotecnologia, Brasília, Brazil. The selected strains were characterized by morphological (microscopy, biochemical (SDS-PAGE 10% and molecular (PCR methods. Six B. thuringiensis strains were identified as mosquito-toxic after the selective bioassays. None of the strains produced the expected PCR products for detection of cry4, cry11 and cyt1A genes. These results indicate that the activity of mosquitocidal Brazilian strains are not related with Cry4, Cry11 or Cyt proteins, so they could be used as an alternative bioinsecticide against mosquitoes.Neste trabalho foram realizados testes de patogenicidade com 210 estirpes de Bacillus thuringiensis contra larvas de Aedes aegypti e Culex quinquefasciatus, a fim de se determinar as mais eficazes. Estas estirpes foram isoladas de diversas regiões do Brasil e estão armazenadas na coleção de Bacillus spp. da Embrapa Recursos Genéticos e Biotecnologia. As estirpes selecionadas foram caracterizadas por métodos morfológicos (microscopia, bioquímicos (SDS-PAGE 10% e moleculares (Reação em Cadeia da Polimerase. Foram selecionadas seis estirpes entomopatogênicas de Bacillus thuringiensis. Nenhuma das estirpes de Bacillus thuringiensis apresentou produtos de PCR esperados para a detecção dos genes cry4, cry11 e cyt1A. A patogenicidade das estirpes não está associada à presença das toxinas Cry4, Cry11 ou Cyt, assim, essas estirpes poderão ser utilizadas para a formatação de um bioinseticida alternativo contra mosquitos.

  17. Trifluoromethylphenyl amides as novel insecticides and fungicides

    Science.gov (United States)

    Because of increased resistance to insecticides in arthropods, it is necessary to identify new chemicals that may have novel modes of action. Following an extensive literature search for compounds with insecticidal and mosquito repellent activity, we have designed and synthesized a set of 20 trifluo...

  18. Induction of rapid and selective cell necrosis in Drosophila using Bacillus thuringiensis Cry toxin and its silkworm receptor.

    Science.gov (United States)

    Obata, Fumiaki; Tanaka, Shiho; Kashio, Soshiro; Tsujimura, Hidenobu; Sato, Ryoichi; Miura, Masayuki

    2015-07-08

    Genetic ablation of target cells is a powerful tool to study the origins and functions of cells, tissue regeneration, or pathophysiology in a human disease model in vivo. Several methods for selective cell ablation by inducing apoptosis have been established, using exogenous toxins or endogenous proapoptotic genes. However, their application is limited to cells with intact apoptotic machinery. Herein, we established a method for inducing rapid and selective cell necrosis by the pore-forming bacterial toxin Cry1Aa, which is specifically active in cells expressing the Cry1Aa receptor (CryR) derived from the silkworm Bombyx mori. We demonstrated that overexpressing CryR in Drosophila melanogaster tissues induced rapid cell death of CryR-expressing cells only, in the presence of Cry1Aa toxin. Cry/CryR system was effective against both proliferating cells in imaginal discs and polyploid postmitotic cells in the fat body. Live imaging analysis of cell ablation revealed swelling and subsequent osmotic lysis of CryR-positive cells after 30 min of incubation with Cry1Aa toxin. Osmotic cell lysis was still triggered when apoptosis, JNK activation, or autophagy was inhibited, suggesting that Cry1Aa-induced necrotic cell death occurred independently of these cellular signaling pathways. Injection of Cry1Aa into the body cavity resulted in specific ablation of CryR-expressing cells, indicating the usefulness of this method for in vivo cell ablation. With Cry toxins from Bacillus thuringiensis, we developed a novel method for genetic induction of cell necrosis. Our system provides a "proteinous drill" for killing target cells through physical injury of the cell membrane, which can potentially be used to ablate any cell type in any organisms, even those that are resistant to apoptosis or JNK-dependent programmed cell death.

  19. Potencial de Bacillus thuringiensis israelensis Berliner no controle de Aedes aegypti Potential of Bacillus thuringiensis israelensis Berliner for controlling Aedes aegypti

    Directory of Open Access Journals (Sweden)

    Ricardo Antonio Polanczyk

    2003-12-01

    Full Text Available Relata-se a importância da bactéria entomopatogênica Bacillus thuringiensis israelensis para o controle de Aedes aegypti. São abordados a utilização e potencial de B. thuringiensis israelensis contra o mosquito vetor da dengue. Outros aspectos são discutidos como a evolução da resistência dos insetos em relação aos inseticidas químicos e as vantagens e desvantagens do controle microbiano como estratégia de controle. É dada ênfase à importância da utilização desta bactéria no Brasil como alternativa para resolver o problema em questão sem afetar o ambiente, o homem e outros vertebrados nas áreas de risco.The importance of the entomopathogenic bacterium Bacillus thuringiensis israelensis in the control of Aedes aegypti is presented. The use and potential of B. thuringiensis israelensis against the mosquito vector of dengue fever is described. Other aspects such as insect's resistance development against chemicals and advantages and constraints of using microbial control are discussed. Emphasis is given to the importance of the use of this bacterium in Brazil, which could contribute significantly to solving the mosquito problem without affecting the environment, humans and others invertebrate organisms in critical regions.

  20. expVIP: a Customizable RNA-seq Data Analysis and Visualization Platform.

    Science.gov (United States)

    Borrill, Philippa; Ramirez-Gonzalez, Ricardo; Uauy, Cristobal

    2016-04-01

    The majority of transcriptome sequencing (RNA-seq) expression studies in plants remain underutilized and inaccessible due to the use of disparate transcriptome references and the lack of skills and resources to analyze and visualize these data. We have developed expVIP, an expression visualization and integration platform, which allows easy analysis of RNA-seq data combined with an intuitive and interactive interface. Users can analyze public and user-specified data sets with minimal bioinformatics knowledge using the expVIP virtual machine. This generates a custom Web browser to visualize, sort, and filter the RNA-seq data and provides outputs for differential gene expression analysis. We demonstrate expVIP's suitability for polyploid crops and evaluate its performance across a range of biologically relevant scenarios. To exemplify its use in crop research, we developed a flexible wheat (Triticum aestivum) expression browser (www.wheat-expression.com) that can be expanded with user-generated data in a local virtual machine environment. The open-access expVIP platform will facilitate the analysis of gene expression data from a wide variety of species by enabling the easy integration, visualization, and comparison of RNA-seq data across experiments. © 2016 American Society of Plant Biologists. All Rights Reserved.

  1. Bacillus thuringiensis (Bt) transgenic crop: an environment friendly insect-pest management strategy.

    Science.gov (United States)

    Kumar, Suresh; Chandra, Amaresh; Pandey, K C

    2008-09-01

    Introduction of DDT (dichloro-diphenyl-trichloroethane) and following move towards indiscriminate use of synthetic chemical insecticides led to the contamination of water and food sources, poisoning of non-target beneficial insects and development of insect-pests resistant to the chemical insecticides. Increased public concems about the adverse environmental effects of indiscriminate use of chemical insecticides prompted search of altemative methods for insect-pest control. One of the promising alternatives has been the use of biological control agents. There is well-documented history of safe application of Bt (B. thuringiensis, a gram positive soil bacterium) as effective biopesticides and a number of reports of expression of delta-endotoxin gene(s) in crop plants are available. Only a few insecticidal sprays are required on Bt transgenic crops, which not only save cost and time, but also reduce health risks. Insects exhibit remarkable ability to develop resistance to different insecticidal compounds, which raises concern about the unsystematic use of Bt transgenic technology also. Though resistance to Bt products among insect species under field conditions has been rare, laboratory studies show that insects are capable of developing high levels of resistance to one ormore Cry proteins. Now it is generally agreed that 'high-dose/refuge strategy' is the most promising and practical approach to prolong the effectiveness of Bt toxins. Although manybiosafety concerns, ethical and moral issues exist, area under Bt transgenic crops is rapidly increasing and they are cultivated on more than 32 million hectares world over Even after reservation of European Union (EU) for acceptance of geneticaly modified (GM) crops, 6 out of 25 countries have already adopted Bt crops and many otherindustrial countries will adopt Bt transgenic crops in near future. While the modem biotechnology has been recognized to have a great potential for the promotion of human well-being, adoption

  2. Monitoring resistance to Bacillus thuringiensis subsp. israelensis in the field by performing bioassays with each Cry toxin separately

    Directory of Open Access Journals (Sweden)

    Guillaume Tetreau

    2013-11-01

    Full Text Available Bacillus thuringiensis subsp. israelensis (Bti is increasingly used worldwide for mosquito control and is the only larvicide used in the French Rhône-Alpes region since decades. The artificial selection of mosquitoes with field-persistent Bti collected in breeding sites from this region led to a moderate level of resistance to Bti, but to relatively high levels of resistance to individual Bti Cry toxins. Based on this observation, we developed a bioassay procedure using each Bti Cry toxin separately to detect cryptic Bti-resistance evolving in field mosquito populations. Although no resistance to Bti was detected in none of the three mosquito species tested (Aedes rusticus, Aedes sticticus and Aedes vexans, an increased tolerance to Cry4Aa (3.5-fold and Cry11Aa toxins (8-fold was found in one Ae. sticticus population compared to other populations of the same species, suggesting that resistance to Bti may be arising in this population. This study confirms previous works showing a lack of Bti resistance in field mosquito populations treated for decades with this bioinsecticide. It also provides a first panorama of their susceptibility status to individual Bti Cry toxins. In combination with bioassays with Bti, bioassays with separate Cry toxins allow a more sensitive monitoring of Bti-resistance in the field.

  3. NCBI nr-aa BLAST: CBRC-PMAR-01-0052 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-PMAR-01-0052 ref|YP_893536.1| spore germination protein [Bacillus thuringiensi...s str. Al Hakam] gb|ABK84029.1| spore germination protein [Bacillus thuringiensis str. Al Hakam] YP_893536.1 0.45 25% ...

  4. Expression of phosphorylated cAMP response element binding protein (p-CREB) in bladder afferent pathways in VIP-/- mice with cyclophosphamide (CYP)-induced cystitis

    DEFF Research Database (Denmark)

    Jensen, Dorthe G; Studeny, Simon; May, Victor

    2008-01-01

    The expression of phosphorylated cAMP response element binding protein (p-CREB) in dorsal root ganglia (DRG) with and without cyclophosphamide (CYP)-induced cystitis (150 mg/kg, i.p; 48 h) was determined in VIP(-/-) and wild-type (WT) mice. p-CREB immunoreactivity (IR) was determined in bladder...... (Fast blue) afferent cells. Nerve growth factor (NGF) bladder content was determined by enzyme-linked immunosorbent assays. Basal expression of p-CREB-IR in DRG of VIP(-/-) mice was (p DRG compared to WT mice. CYP treatment in WT mice increased (p ...-CREB-IR in L1, L2, L5-S1 DRG. CYP treatment in VIP(-/-) mice (p DRG compared to WT with CYP. In WT mice, bladder afferent cells (20-38%) in DRG expressed p-CREB-IR under basal conditions. With CYP, p-CREB-IR increased in bladder afferent cells (60...

  5. Detector location selection based on VIP analysis in near-infrared detection of dural hematoma

    Directory of Open Access Journals (Sweden)

    Qiuming Sun

    2018-03-01

    Full Text Available Detection of dural hematoma based on multi-channel near-infrared differential absorbance has the advantages of rapid and non-invasive detection. The location and number of detectors around the light source are critical for reducing the pathological characteristics of the prediction model on dural hematoma degree. Therefore, rational selection of detector numbers and their distances from the light source is very important. In this paper, a detector position screening method based on Variable Importance in the Projection (VIP analysis is proposed. A preliminary modeling based on Partial Least Squares method (PLS for the prediction of dural position μa was established using light absorbance information from 30 detectors located 2.0–5.0 cm from the light source with a 0.1 cm interval. The mean relative error (MRE of the dural position μa prediction model was 4.08%. After VIP analysis, the number of detectors was reduced from 30 to 4 and the MRE of the dural position μa prediction was reduced from 4.08% to 2.06% after the reduction in detector numbers. The prediction model after VIP detector screening still showed good prediction of the epidural position μa. This study provided a new approach and important reference on the selection of detector location in near-infrared dural hematoma detection. Keywords: Detector location screening, Epidural hematoma detection, Variable importance in the projection

  6. The Fermi LAT Very Important Project (VIP) List of Active Galactic Nuclei

    Science.gov (United States)

    Thompson, David J.; Fermi Large Area Telescope Collaboration

    2018-01-01

    Using nine years of Fermi Gamma-ray Space Telescope Large Area Telescope (LAT) observations, we have identified 30 projects for Active Galactic Nuclei (AGN) that appear to provide strong prospects for significant scientific advances. This Very Important Project (VIP) AGN list includes AGNs that have good multiwavelength coverage, are regularly detected by the Fermi LAT, and offer scientifically interesting timing or spectral properties. Each project has one or more LAT scientists identified who are actively monitoring the source. They will be regularly updating the LAT results for these VIP AGNs, working together with multiwavelength observers and theorists to maximize the scientific return during the coming years of the Fermi mission. See https://confluence.slac.stanford.edu/display/GLAMCOG/VIP+List+of+AGNs+for+Continued+Study

  7. Transgenic cotton expressing Cry10Aa toxin confers high resistance to the cotton boll weevil.

    Science.gov (United States)

    Ribeiro, Thuanne Pires; Arraes, Fabricio Barbosa Monteiro; Lourenço-Tessutti, Isabela Tristan; Silva, Marilia Santos; Lisei-de-Sá, Maria Eugênia; Lucena, Wagner Alexandre; Macedo, Leonardo Lima Pepino; Lima, Janaina Nascimento; Santos Amorim, Regina Maria; Artico, Sinara; Alves-Ferreira, Márcio; Mattar Silva, Maria Cristina; Grossi-de-Sa, Maria Fatima

    2017-08-01

    Genetically modified (GM) cotton plants that effectively control cotton boll weevil (CBW), which is the most destructive cotton insect pest in South America, are reported here for the first time. This work presents the successful development of a new GM cotton with high resistance to CBW conferred by Cry10Aa toxin, a protein encoded by entomopathogenic Bacillus thuringiensis (Bt) gene. The plant transformation vector harbouring cry10Aa gene driven by the cotton ubiquitination-related promoter uceA1.7 was introduced into a Brazilian cotton cultivar by biolistic transformation. Quantitative PCR (qPCR) assays revealed high transcription levels of cry10Aa in both T 0 GM cotton leaf and flower bud tissues. Southern blot and qPCR-based 2 -ΔΔCt analyses revealed that T 0 GM plants had either one or two transgene copies. Quantitative and qualitative analyses of Cry10Aa protein expression showed variable protein expression levels in both flower buds and leaves tissues of T 0 GM cotton plants, ranging from approximately 3.0 to 14.0 μg g -1 fresh tissue. CBW susceptibility bioassays, performed by feeding adults and larvae with T 0 GM cotton leaves and flower buds, respectively, demonstrated a significant entomotoxic effect and a high level of CBW mortality (up to 100%). Molecular analysis revealed that transgene stability and entomotoxic effect to CBW were maintained in T 1 generation as the Cry10Aa toxin expression levels remained high in both tissues, ranging from 4.05 to 19.57 μg g -1 fresh tissue, and the CBW mortality rate remained around 100%. In conclusion, these Cry10Aa GM cotton plants represent a great advance in the control of the devastating CBW insect pest and can substantially impact cotton agribusiness. © 2017 The Authors. Plant Biotechnology Journal published by Society for Experimental Biology and The Association of Applied Biologists and John Wiley & Sons Ltd.

  8. VIP A B C. Vacuum Insulation Panels Applied in Building Constructions

    Energy Technology Data Exchange (ETDEWEB)

    Tenpierik, M.J.

    2010-02-01

    Due to sustainability and due to international treaties, it is desired and required to reduce greenhouse gas emissions drastically. One contributor to these emissions is the burning of fossil fuels for generating power and electricity to be used in and for buildings. Buildings and building-related processes are responsible for about 40% of the primary energy consumption in the European Union. More than half of this energy is applied for heating systems in dwellings and commercial buildings. The European Union therefore has laid down new energy performance requirements for buildings in the European Directive on the Energy Performance of Buildings. Moreover, a reduction of energy losses of buildings during their occupational phase is important for facilitating the implementation of sustainable energy sources in the built environment. Increasing the insulation value of the envelope of buildings may contribute to this reduction of primary energy use. Two strategies can be followed. The first strategy is to increase the thickness of the thermal insulation layer. Until recently, this strategy has primarily been adopted. If, however, German or Swiss Passivhaus standard is applied, the thickness of this insulation layer would increase to beyond 30 cm, resulting in very thick building enclosures. The second, more innovative, strategy for reducing energy losses through the building skin would be the application of more effective thermal insulators. One such more effective thermal insulator is a vacuum insulation panel, abbreviated as VIP. A VIP consists of an open-celled core material which is evacuated and then tightly sealed into a barrier envelope to maintain this vacuum. The vacuum inside the pores of the core material reduces the thermal conductivity of the product significantly, as a result of which the thickness of the insulation layer can be reduced to obtain a certain performance. This reduction of thickness is among the most promising features for large

  9. 40 CFR 174.532 - Bacillus thuringiensis eCry3.1Ab protein in corn; temporary exemption from the requirement of a...

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 23 2010-07-01 2010-07-01 false Bacillus thuringiensis eCry3.1Ab protein in corn; temporary exemption from the requirement of a tolerance. 174.532 Section 174.532 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) PESTICIDE PROGRAMS PROCEDURES AND REQUIREMENTS FOR PLANT-INCORPORATED PROTECTANTS...

  10. AAS 228: Day 3 afternoon

    Science.gov (United States)

    Kohler, Susanna

    2016-06-01

    black hole interiors, we cant learn whats in them via observation. Instead, we rely on mathematical descriptions of what their interiors look like. Henry and collaborators have developed a new coordinate-invariant depiction of the structure of black holes that reframes how we think about visualize their interiors. More information can be found in their paper here.The final press conference presentation was given by Maria Dainotti (Stanford University) on the subject of using gamma-ray bursts (GRBs) as standard candles. Standard candles are astrophysical sources that have known luminosities. We can use the light we observe from standard candles to estimate their distance, making them useful tools for testing cosmological models.Type Ia supernovae (which always have roughly the same source brightness) are a typical example of a standard candle, and led to the discovery of dark energy in the 1990s. But if GRBs could be used in a similar way, they would be a much more powerful tool: theyre visible out to significantly further distances because theyre so bright. Dainotti and collaborators are analyzing a subset of GRBs from Swift observations that they term gold GRBs. By examining the correlations between parameters of these gold GRBs, Dainotti attempts to understand whether they can be used as standard candles.Plenary Session From the First Stars and Galaxies to the Epoch of Reionization: 20 Years of Computational Progress (by Gourav Khullar)Mike Normans (University of California San Diego) plenary talk concluded the AAS Plenary sessions with a treatise on the current generation of simulations that aim to characterize the first stars and galaxies that formed in the Universe. Last plenary talk for today. #aas228pic.twitter.com/uTaWt6Qi00 Daniel Caton (@DanielCaton) June 15, 2016Mike has been working in the field for a long time, and his collaborators work on the Enzo simulations is very well known. This talk was centered around some of the latest Enzo results, with a special

  11. PACAP and VIP inhibit the invasiveness of glioblastoma cells exposed to hypoxia through the regulation of HIFs and EGFR expression

    Directory of Open Access Journals (Sweden)

    Grazia eMaugeri

    2016-05-01

    Full Text Available Pituitary adenylate cyclase-activating polypeptide (PACAP and vasoactive intestinal peptide (VIP through the binding of vasoactive intestinal peptide receptors (VIPRs, perform a wide variety of effects in human cancers, including glioblastoma multiforme (GBM. This tumor is characterized by extensive areas of hypoxia, which triggers the expression of hypoxia-inducible factors (HIFs. HIFs not only mediate angiogenesis but also tumor cell migration and invasion. Furthermore, HIFs activation is linked to epidermal growth factor receptor (EGFR overexpression. Previous studies have shown that VIP interferes with the invasive nature of gliomas by regulating cell migration. However, the role of VIP family members in GBM infiltration under low oxygen tension has not been clarified yet. Therefore, in the present study we have investigated, for the first time, the molecular mechanisms involved in the anti-invasive effect of PACAP or VIP in U87MG glioblastoma cells exposed to hypoxia induced by treatment with desferrioxamine (DFX. The results suggest that either PACAP or VIP exert an anti-infiltrative effect under low oxygen tension by modulating HIFs and EGFR expression, key elements involved in cell migration and angiogenesis. These peptides act through the inhibition of PI3K/Akt and MAPK/ERK signaling pathways, which are known to have a crucial role in HIFs regulation. In conclusion, the modulation of hypoxic event and the anti-invasive effect exerted by some VIP family members might open new insights in the therapeutic approach to GBM.

  12. VIP as a potential therapeutic agent in gram negative sepsis.

    Science.gov (United States)

    Ibrahim, Hiba; Barrow, Paul; Foster, Neil

    2012-12-01

    Gram negative sepsis remains a high cause of mortality and places a great burden on public health finance in both the developed and developing world. Treatment of sepsis, using antibiotics, is often ineffective since pathology associated with the disease occurs due to dysregulation of the immune system (failure to return to steady state conditions) which continues after the bacteria, which induced the immune response, have been cleared. Immune modulation is therefore a rational approach to the treatment of sepsis but to date no drug has been developed which is highly effective, cheap and completely safe to use. One potential therapeutic agent is VIP, which is a natural peptide and is highly homologous in all vertebrates. In this review we will discuss the effect of VIP on components of the immune system, relevant to gram negative sepsis, and present data from animal models. Furthermore we will hypothesise on how these studies could be improved in future and speculate on the possible different ways in which VIP could be used in clinical medicine.

  13. Enhancement of virulence of bacillus thuringiensis and serratia marcescens by chemicals

    International Nuclear Information System (INIS)

    Khan, K. A.

    2006-01-01

    Studies were conducted on the enhancement of pathogenicity of Bacillus thuringiensis by 1% boric acid against various species of termites. The increase in virulence of Serratia marcescens by 1% potassium chloride or 1% Sodium citrate against the workers of M. championi has also been established. The increase in virulence is confirmed by the enhancement ratio, which are ranging from about 1.5 to 1.8 for Bacillus thuringiensis and 1.3 to 1.6 for Serratia marcescens. It was also noted that 1% boric acid alone was found toxic to various species of termites. However, Potassium chloride and Sodium citrate in a concentration of 1% were non-toxic to the workers of M. championi. (author)

  14. Characterization of a highly toxic strain of Bacillus thuringiensis serovar kurstaki very similar to the HD-73 strain.

    Science.gov (United States)

    Reinoso-Pozo, Yaritza; Del Rincón-Castro, Ma Cristina; Ibarra, Jorge E

    2016-09-01

    The LBIT-1200 strain of Bacillus thuringiensis was recently isolated from soil, and showed a 6.4 and 9.5 increase in toxicity, against Manduca sexta and Trichoplusia ni, respectively, compared to HD-73. However, LBIT-1200 was still highly similar to HD-73, including the production of bipyramidal crystals containing only one protein of ∼130 000 kDa, its flagellin gene sequence related to the kurstaki serotype, plasmid and RepPCR patterns similar to HD-73, no production of β-exotoxin and no presence of VIP genes. Sequencing of its cry gene showed the presence of a cry1Ac-type gene with four amino acid differences, including two amino acid replacements in domain III, compared to Cry1Ac1, which may explain its higher toxicity. In conclusion, the LBIT-1200 strain is a variant of the HD-73 strain but shows a much higher toxicity, which makes this new strain an important candidate to be developed as a bioinsecticide, once it passes other tests, throughout its biotechnological development. © FEMS 2016. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  15. Distinct roles of SOM and VIP interneurons during cortical Up states

    Directory of Open Access Journals (Sweden)

    Garrett T. Neske

    2016-07-01

    Full Text Available During cortical network activity, recurrent synaptic excitation among pyramidal neurons is approximately balanced by synaptic inhibition, which is provided by a vast diversity of inhibitory interneurons. The relative contributions of different interneuron subtypes to inhibitory tone during cortical network activity is not well understood. We previously showed that many of the major interneuron subtypes in mouse barrel cortex are highly active during Up states (Neske et al., 2015; while fast-spiking (FS, parvalbumin (PV-positive cells were the most active interneuron subtype, many non-fast-spiking (NFS, PV-negative interneurons were as active or more active than neighboring pyramidal cells. This suggests that the NFS cells could play a role in maintaining or modulating Up states. Here, using optogenetic techniques, we further dissected the functional roles during Up states of two major NFS, PV-negative interneuron subtypes: somatostatin (SOM-positive cells and vasoactive intestinal peptide (VIP-positive cells. We found that while pyramidal cell excitability during Up states significantly increased when SOM cells were optogenetically silenced, VIP cells did not influence pyramidal cell excitability either upon optogenetic silencing or activation. VIP cells failed to contribute to Up states despite their ability to inhibit SOM cells strongly. We suggest that the contribution of VIP cells to the excitability of pyramidal cells may vary with cortical state.

  16. Atrial natriuretic peptide stimulates salt secretion by shark rectal gland by releasing VIP

    Energy Technology Data Exchange (ETDEWEB)

    Silva, P.; Stoff, J.S.; Solomon, R.J.; Lear, S.; Kniaz, D.; Greger, R.; Epstein, F.H.

    1987-01-01

    Salt secretion by the isolated perfused rectal gland of the spiny dogfish shark, Squalus acanthias, is stimulated by synthetic rat atrial natriuretic peptide (ANP II) as well as extracts of shark heart, but not by 8-bromo-cyclic guanosine 5'-monophosphate. Cardiac peptides have no effect on isolated rectal gland cells or perfused tubules, suggesting that stimulation requires an intact gland. The stimulation of secretion by ANP II is eliminated by maneuvers that block neurotransmitter release. Cardiac peptides stimulate the release of vasoactive intestinal peptide (VIP), known to be present in rectal glands nerves, into the venous effluent of perfused glands in parallel with their stimulation of salt secretion, but the release of VIP induced by ANP II is prevented by perfusion with procaine. VIP was measured by radioimmunoassay. Cardiac peptides thus appear to regulate rectal gland secretion by releasing VIP from neural stores within the gland. It is possible that other physiological effects of these hormones might be explained by an action to enhanced local release of neurotransmitters.

  17. Atrial natriuretic peptide stimulates salt secretion by shark rectal gland by releasing VIP

    International Nuclear Information System (INIS)

    Silva, P.; Stoff, J.S.; Solomon, R.J.; Lear, S.; Kniaz, D.; Greger, R.; Epstein, F.H.

    1987-01-01

    Salt secretion by the isolated perfused rectal gland of the spiny dogfish shark, Squalus acanthias, is stimulated by synthetic rat atrial natriuretic peptide (ANP II) as well as extracts of shark heart, but not by 8-bromo-cyclic guanosine 5'-monophosphate. Cardiac peptides have no effect on isolated rectal gland cells or perfused tubules, suggesting that stimulation requires an intact gland. The stimulation of secretion by ANP II is eliminated by maneuvers that block neurotransmitter release. Cardiac peptides stimulate the release of vasoactive intestinal peptide (VIP), known to be present in rectal glands nerves, into the venous effluent of perfused glands in parallel with their stimulation of salt secretion, but the release of VIP induced by ANP II is prevented by perfusion with procaine. VIP was measured by radioimmunoassay. Cardiac peptides thus appear to regulate rectal gland secretion by releasing VIP from neural stores within the gland. It is possible that other physiological effects of these hormones might be explained by an action to enhanced local release of neurotransmitters

  18. In Silico Modeling and Functional Interpretations of Cry1Ab15 Toxin from Bacillus thuringiensis BtB-Hm-16

    Directory of Open Access Journals (Sweden)

    Sudhanshu Kashyap

    2013-01-01

    Full Text Available The theoretical homology based structural model of Cry1Ab15 δ-endotoxin produced by Bacillus thuringiensis BtB-Hm-16 was predicted using the Cry1Aa template (resolution 2.25 Å. The Cry1Ab15 resembles the template structure by sharing a common three-domain extending conformation structure responsible for pore-forming and specificity determination. The novel structural differences found are the presence of β0 and α3, and the absence of α7b, β1a, α10a, α10b, β12, and α11a while α9 is located spatially downstream. Validation by SUPERPOSE and with the use of PROCHECK program showed folding of 98% of modeled residues in a favourable and stable orientation with a total energy Z-score of −6.56; the constructed model has an RMSD of only 1.15 Å. These increments of 3D structure information will be helpful in the design of domain swapping experiments aimed at improving toxicity and will help in elucidating the common mechanism of toxin action.

  19. Resistance to dual-gene Bt maize in Spodoptera frugiperda: selection, inheritance, and cross-resistance to other transgenic events.

    Science.gov (United States)

    Santos-Amaya, Oscar F; Rodrigues, João V C; Souza, Thadeu C; Tavares, Clébson S; Campos, Silverio O; Guedes, Raul N C; Pereira, Eliseu J G

    2015-12-17

    Transgenic crop "pyramids" producing two or more Bacillus thuringiensis (Bt) toxins active against the same pest are used to delay evolution of resistance in insect pest populations. Laboratory and greenhouse experiments were performed with fall armyworm, Spodoptera frugiperda, to characterize resistance to Bt maize producing Cry1A.105 and Cry2Ab and test some assumptions of the "pyramid" resistance management strategy. Selection of a field-derived strain of S. frugiperda already resistant to Cry1F maize with Cry1A.105 + Cry2Ab maize for ten generations produced resistance that allowed the larvae to colonize and complete the life cycle on these Bt maize plants. Greenhouse experiments revealed that the resistance was completely recessive (Dx = 0), incomplete, autosomal, and without maternal effects or cross-resistance to the Vip3Aa20 toxin produced in other Bt maize events. This profile of resistance supports some of the assumptions of the pyramid strategy for resistance management. However, laboratory experiments with purified Bt toxin and plant leaf tissue showed that resistance to Cry1A.105 + Cry2Ab2 maize further increased resistance to Cry1Fa, which indicates that populations of fall armyworm have high potential for developing resistance to some currently available pyramided maize used against this pest, especially where resistance to Cry1Fa was reported in the field.

  20. Field Trial Performance of Herculex XTRA (Cry34Ab1/Cry35Ab1) and SmartStax (Cry34Ab1/Cry35Ab1 + Cry3Bb1) Hybrids and Soil Insecticides Against Western and Northern Corn Rootworms (Coleoptera: Chrysomelidae).

    Science.gov (United States)

    Johnson, K D; Campbell, L A; Lepping, M D; Rule, D M

    2017-06-01

    Western corn rootworm, Diabrotica virgifera virgifera LeConte (Coleoptera: Chrysomelidae), and northern corn rootworm, Diabrotica barberi Smith and Lawrence (Coleoptera: Chrysomelidae), are important insect pests in corn, Zea mays L. For more than a decade, growers have been using transgenic plants expressing proteins from the bacterium Bacillus thuringiensis (Bt) to protect corn roots from feeding. In 2011, western corn rootworm populations were reported to have developed resistance to Bt hybrids expressing Cry3Bb1 and later found to be cross-resistant to hybrids expressing mCry3A and eCry3.1Ab. The identification of resistance to Cry3 (Cry3Bb1, mCry3A, and eCry3.1Ab) hybrids led to concerns about durability and efficacy of products with single traits and of products containing a pyramid of a Cry3 protein and the binary Bt proteins Cry34Ab1 and Cry35Ab1. From 2012 to 2014, 43 field trials were conducted across the central United States to estimate root protection provided by plants expressing Cry34Ab1/Cry35Ab1 alone (Herculex RW) or pyramided with Cry3Bb1 (SmartStax). These technologies were evaluated with and without soil-applied insecticides to determine if additional management measures provided benefit where Cry3 performance was reduced. Trials were categorized for analysis based on rootworm damage levels on Cry3-expressing hybrids and rootworm feeding pressure within each trial. Across scenarios, Cry34Ab1/Cry35Ab1 hybrids provided excellent root protection. Pyramided traits provided greater root and yield protection than non-Bt plus a soil-applied insecticide, and only in trials where larval feeding pressure exceeded two nodes of damage did Cry34Ab1/Cry35Ab1 single-trait hybrids and pyramided hybrids show greater root protection from the addition of soil-applied insecticides. © 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.

  1. Transgenic Bt Corn, Soil Insecticide, and Insecticidal Seed Treatment Effects on Corn Rootworm (Coleoptera: Chrysomelidae) Beetle Emergence, Larval Feeding Injury, and Corn Yield in North Dakota.

    Science.gov (United States)

    Calles-Torrez, Veronica; Knodel, Janet J; Boetel, Mark A; Doetkott, Curt D; Podliska, Kellie K; Ransom, Joel K; Beauzay, Patrick; French, B Wade; Fuller, Billy W

    2018-02-09

    Northern, Diabrotica barberi Smith & Lawrence (Coleoptera: Chrysomelidae), and western, Diabrotica virgifera virgifera LeConte (Coleoptera: Chrysomelidae), corn rootworms are economic pests of corn, Zea mays L. in North America. We measured the impacts of corn hybrids incorporated with Cry3Bb1, Cry34/35Ab1, and pyramided (Cry3Bb1 + Cry34/35Ab1) Bacillus thuringiensis Berliner (Bt) proteins, tefluthrin soil insecticide, and clothianidin insecticidal seed treatment on beetle emergence, larval feeding injury, and corn yield at five locations from 2013 to 2015 in eastern North Dakota. In most cases, emergence was significantly lower in Bt-protected corn than in non-Bt corn hybrids. Exceptions included Wyndmere, ND (2013), where D. barberi emergence from Cry34/35Ab1 plots was not different from that in the non-Bt hybrid, and Arthur, ND (2013), where D. v. virgifera emergence from Cry3Bb1 plots did not differ from that in the non-Bt hybrid. Bt hybrids generally produced increased grain yield compared with non-Bt corn where rootworm densities were high, and larval root-feeding injury was consistently lower in Bt-protected plots than in non-Bt corn. The lowest overall feeding injury and emergence levels occurred in plots planted with the Cry3Bb1 + Cry34/35Ab1 hybrid. Time to 50% cumulative emergence of both species was 5-7 d later in Bt-protected than in non-Bt hybrids. Tefluthrin and clothianidin were mostly inconsequential in relation to beetle emergence and larval root injury. Our findings could suggest that some North Dakota populations could be in early stages of increased tolerance to some Bt toxins; however, Bt corn hybrids currently provide effective protection against rootworm injury in eastern North Dakota. © The Author(s) 2017. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  2. Preliminary investigation on the effects of biological and synthetic insecticides on large white butterfly (Pieris brassicae L. larvae

    Directory of Open Access Journals (Sweden)

    Klokočar-Šmit Zlata D.

    2007-01-01

    Full Text Available Control of cabbage pests is oriented towards the use of efficient but high-risk insecticides, some of them being endocrine disruptors. Biopesticides are more environment-friendly, operator-and consumers-safe, but they have low initial toxicity, low efficacy to advanced larval stages, and they require certain knowledge of pest and host biology. In our laboratory experiments we have investigated the effects of formulated synthetic pyrethroid cypermethrin (0.3 l/ha and biological products - formulations based on Bacillus thuringiensis subsp. kurstaki (2 and 3/ha and Spinosad (0.1 l/ha - on large white butterfly (Pieris brassicae L. larvae-instars 2, 3, 4 and 5. The effect of insecticides was inversely proportional to larval instars. Btk effect could be improved if tank-mixed with cypermethrin. The mixing of ready-made products allows a reduction 3 and 6 times compared with the recommended dose, still obtaining satisfactory results. Rate of leaf damage was reduced when tank mixtures were used. Use of two products in mixture would be of significance especially for control of advanced late instars late in season, when Btk action alone is insufficient. Spinosad was effective in inducing mortality and reducing leaf damage by all larval instars, therefore we assume that the dose could be reduced. Feeding rate and mortality are equally important parameters when assessing biopesticide efficacy. This strategy should also reduce the possibility of inducing resistance in pest population. It also tends to reduce the residues in commodities and is good solution in production of hygienic and health safe food.

  3. Underground test of quantum mechanics - the VIP2 experiment arXiv

    CERN Document Server

    Marton, Johann; Bassi, A.; Bazzi, M.; Bertolucci, S.; Berucci, C.; Bragadireanu, M.; Cargnelli, M.; Clozza, A.; Curceanu, C.; De Paolis, L.; Di Matteo, S.; Donadi, S.; Egger, J.-P.; Guaraldo, C.; Iliescu, M.; Laubenstein, M.; Milotti, E.; Pichler, A.; Pietreanu, D.; Piscicchia, K.; Scordo, A.; Shi, H.; Sirghi, D.; Sirghi, F.; Sperandio, L.; Vazquez-Doce, O.; Widmann, E.; Zmeskal, J.

    We are experimentally investigating possible violations of standard quantum mechanics predictions in the Gran Sasso underground laboratory in Italy. We test with high precision the Pauli Exclusion Principle and the collapse of the wave function (collapse models). We present our method of searching for possible small violations of the Pauli Exclusion Principle (PEP) for electrons, through the search for anomalous X-ray transitions in copper atoms, produced by fresh electrons (brought inside the copper bar by circulating current) which can have the probability to undergo Pauli-forbidden transition to the 1 s level already occupied by two electrons and we describe the VIP2 (VIolation of PEP) experiment under data taking at the Gran Sasso underground laboratories. In this paper the new VIP2 setup installed in the Gran Sasso underground laboratory will be presented. The goal of VIP2 is to test the PEP for electrons with unprecedented accuracy, down to a limit in the probability that PEP is violated at the level of...

  4. The Vip1 inositol polyphosphate kinase family regulates polarized growth and modulates the microtubule cytoskeleton in fungi.

    Directory of Open Access Journals (Sweden)

    Jennifer Pöhlmann

    2014-09-01

    Full Text Available Microtubules (MTs are pivotal for numerous eukaryotic processes ranging from cellular morphogenesis, chromosome segregation to intracellular transport. Execution of these tasks requires intricate regulation of MT dynamics. Here, we identify a new regulator of the Schizosaccharomyces pombe MT cytoskeleton: Asp1, a member of the highly conserved Vip1 inositol polyphosphate kinase family. Inositol pyrophosphates generated by Asp1 modulate MT dynamic parameters independent of the central +TIP EB1 and in a dose-dependent and cellular-context-dependent manner. Importantly, our analysis of the in vitro kinase activities of various S. pombe Asp1 variants demonstrated that the C-terminal phosphatase-like domain of the dual domain Vip1 protein negatively affects the inositol pyrophosphate output of the N-terminal kinase domain. These data suggest that the former domain has phosphatase activity. Remarkably, Vip1 regulation of the MT cytoskeleton is a conserved feature, as Vip1-like proteins of the filamentous ascomycete Aspergillus nidulans and the distantly related pathogenic basidiomycete Ustilago maydis also affect the MT cytoskeleton in these organisms. Consistent with the role of interphase MTs in growth zone selection/maintenance, all 3 fungal systems show aspects of aberrant cell morphogenesis. Thus, for the first time we have identified a conserved biological process for inositol pyrophosphates.

  5. Vasoactive intestinal polypeptide (VIP) tissue distribution in the rat as measured by radioimmunoassay and by radioreceptorassay

    International Nuclear Information System (INIS)

    Besson, J.; Dupont, C.; Laburthe, M.; Bataille, D.; Rosselin, G.

    1977-01-01

    A new radioimmunoassay which allows the measurement of the rat vasoactive intestinal polypeptide, was performed. VIP is present in the whole digestive tract of rat, mainly between the duodenum and the colon. 1.5% of the total VIP is present in brain. The VIP-like immunoreactivity appears to correspond to biologically active molecule since a radioreceptorassay using liver plasma membranes as the target tissue, gives the same results as the radioimmunoassay [fr

  6. Fulltext PDF

    Indian Academy of Sciences (India)

    Unknown

    stressed seedlings of an indica rice. 25. Bacillus thuringiensis. Cloning and characterization of an insecticidal crystal protein gene from Bacillus thuringiensis subspecies kenyae. 5. Bateson, William. William Bateson: a biologist ahead of his time. 49.

  7. Dominant negative phenotype of Bacillus thuringiensis Cry1Ab, Cry11Aa and Cry4Ba mutants suggest hetero-oligomer formation among different Cry toxins.

    NARCIS (Netherlands)

    Carmona, D.; Rodriguez-Almazan, C.; Munoz-Garay, C.; Portugal, L.; Perez, C.; Maagd, de R.A.; Bakker, P.; Soberon, M.; Bravo, A.

    2011-01-01

    Background - Bacillus thuringiensis Cry toxins are used worldwide in the control of different insect pests important in agriculture or in human health. The Cry proteins are pore-forming toxins that affect the midgut cell of target insects. It was shown that non-toxic Cry1Ab helix a-4 mutants had a

  8. In vitro autoradiographic localization of vasoactive intestinal peptide (VIP) binding sites in the rat central nervous system

    International Nuclear Information System (INIS)

    Besson, J.; Dussaillant, M.; Marie, J.C.; Rostene, W.; Rosselin, G.

    1984-01-01

    This paper describes the autoradiographic distribution of VIP binding sites in the rat central nervous system using monoiodinated 125I-labeled VIP. High densities of VIP binding sites are observed in the granular layer of the dorsal dentate gyrus of the hippocampus, the basolateral amygdaloid nucleus, the dorsolateral and median geniculate nuclei of the thalamus as well as in the ventral part of the hypothalamic dorsomedial nucleus

  9. Chemical modulators of the innate immune response alter gypsy moth larval susceptibility to Bacillus thuringiensis

    Directory of Open Access Journals (Sweden)

    Broderick Nichole A

    2010-04-01

    Full Text Available Abstract Background The gut comprises an essential barrier that protects both invertebrate and vertebrate animals from invasion by microorganisms. Disruption of the balanced relationship between indigenous gut microbiota and their host can result in gut bacteria eliciting host responses similar to those caused by invasive pathogens. For example, ingestion of Bacillus thuringiensis by larvae of some species of susceptible Lepidoptera can result in normally benign enteric bacteria exerting pathogenic effects. Results We explored the potential role of the insect immune response in mortality caused by B. thuringiensis in conjunction with gut bacteria. Two lines of evidence support such a role. First, ingestion of B. thuringiensis by gypsy moth larvae led to the depletion of their hemocytes. Second, pharmacological agents that are known to modulate innate immune responses of invertebrates and vertebrates altered larval mortality induced by B. thuringiensis. Specifically, Gram-negative peptidoglycan pre-treated with lysozyme accelerated B. thuringiensis-induced killing of larvae previously made less susceptible due to treatment with antibiotics. Conversely, several inhibitors of the innate immune response (eicosanoid inhibitors and antioxidants increased the host's survival time following ingestion of B. thuringiensis. Conclusions This study demonstrates that B. thuringiensis infection provokes changes in the cellular immune response of gypsy moth larvae. The effects of chemicals known to modulate the innate immune response of many invertebrates and vertebrates, including Lepidoptera, also indicate a role of this response in B. thuringiensis killing. Interactions among B. thuringiensis toxin, enteric bacteria, and aspects of the gypsy moth immune response may provide a novel model to decipher mechanisms of sepsis associated with bacteria of gut origin.

  10. Current research efforts with Bacillus thuringiensis

    Science.gov (United States)

    Normand R. Dubois

    1991-01-01

    The bioassay of 260 strains of Bacillus thuringiensis (Bt) and 70 commercial preparations show that regression coefficient estimates may be as critical as LC5O estimates when evaluating them for future consideration.

  11. Survival and conjugal transfer between Bacillus thuringiensis strains in aquatic environment Sobrevivência e conjugação de Bacillus thuringiensis em ambiente aquático

    Directory of Open Access Journals (Sweden)

    Luciana Furlaneto

    2000-10-01

    Full Text Available Field and laboratory studies were conducted to assess the survival of cells and spores and plasmid transfer between Bacillus thuringienis strains in aquatic environment. Results indicated that cells and spores of B. thuringiensis can survive for 10 days in water, without altering their number. The sporulation process began after 12-15 hours of inoculation of water. B. thuringiensis was able to transfer conjugative plasmids in the aquatic environment.O presente trabalho é um estudo sobre a sobrevivência e a conjugação de linhagens de Bacillus thuringiensis em água. Os experimentos conduzidos no laboratório mostram que as células e os esporos de B. thuringiensis podem persistir pelo menos 10 dias na água. A esporulação inicia-se 12-15 horas após a inoculação. O processo de conjugação foi demonstrado em diferentes ambientes aquáticos, tanto em condições de laboratório quanto no meio ambiente.

  12. Examination of the gastrointestinal tract using the tussian X-ray contrast substance BAR-VIPS based on barium sulfate

    International Nuclear Information System (INIS)

    Ratobyl'skij, G.V.; Kaluzhskij, A.A.

    1997-01-01

    Results of X-ray studies of gastrointestinal tract using domestic X-ray contrast preparation based on barium sulfate BAR-VIPS developed by VIPS-MED company (t. Fryazino, Moscow region). Testing of the preparation has shown that BAR-VIPS permits to diagnose every pathological changes in esophagus, stomach, rectum, small intestine as well as to diagnose successfully large intestine diseases

  13. Use of endophytic diazotrophic bacteria as a vector to express the cry3A gene from Bacillus thuringiensis

    Directory of Open Access Journals (Sweden)

    Salles Joana Falcão

    2000-01-01

    Full Text Available The goal of this study was to evaluate the potential of endophytic diazotrophic bacteria as a vector to express a cry gene from Bacillus thuringiensis, envisaging the control of pests that attack sugarcane plants. The endophytic nitrogen-fixing bacteria Gluconacetobacter diazotrophicus strain BR11281 and Herbaspirillum seropedicae strain BR11335 were used as models. The cry3A gene was transferred by conjugation using a suicide plasmid and the recombinant strains were selected by their ability to fix nitrogen in semi-solid N-free medium. The presence of the cry gene was detected by Southern-blot using an internal fragment of 1.0 kb as a probe. The production of delta-endotoxin by the recombinant H. seropedicae strain was detected by dot blot while for G. diazotrophicus the Western-blot technique was used. In both cases, a specific antibody raised against the B. thuringiensis toxin was applied. The delta-endotoxin production showed by the G. diazotrophicus recombinant strain was dependent on the nitrogen fixing conditions since the cry3A gene was fused to a nif promoter. In the case of H. seropedicae the delta-endotoxin expression was not affected by the promoter (rhi used. These results suggest that endophytic diazotrophic bacteria can be used as vectors to express entomopathogenic genes envisaging control of sugarcane pests.

  14. Marketing plan of Vips Jeans

    OpenAIRE

    Marzá Mallol, Adrià

    2015-01-01

    Treball Final de Grau en Administració d'Empreses. Codi: AE1049. Curs: 2014/2015 The following work shows the Marketing Plan of Vips Jeans, this company is specialized in the commercialization of fashion-forward. His shop in the street ‘En medio’ in Castellón sells clothing, shoes and accessories of the best current brands. This company has been operating since 1988 and their main Competitive Advantage is the social prestige among Castellón consumers, acquired during these 26 ...

  15. Computational and biological characterization of fusion proteins of two insecticidal proteins for control of insect pests.

    Science.gov (United States)

    Javaid, Shaista; Naz, Sehrish; Amin, Imran; Jander, Georg; Ul-Haq, Zaheer; Mansoor, Shahid

    2018-03-19

    Sucking pests pose a serious agricultural challenge, as available transgenic technologies such as Bacillus thuringiensis crystal toxins (Bt) are not effective against them. One approach is to produce fusion protein toxins for the control of these pests. Two protein toxins, Hvt (ω-atracotoxin from Hadronyche versuta) and onion leaf lectin, were translationally fused to evaluate the negative effects of fusion proteins on Phenacoccus solenopsis (mealybug), a phloem-feeding insect pest. Hvt was cloned both N-terminally (HL) and then C-terminally (LH) in the fusion protein constructs, which were expressed transiently in Nicotiana tabacum using a Potato Virus X (PVX) vector. The HL fusion protein was found to be more effective against P. solenopsis, with an 83% mortality rate, as compared to the LH protein, which caused 65% mortality. Hvt and lectin alone caused 42% and 45%, respectively, under the same conditions. Computational studies of both fusion proteins showed that the HL protein is more stable than the LH protein. Together, these results demonstrate that translational fusion of two insecticidal proteins improved the insecticidal activity relative to each protein individually and could be expressed in transgenic plants for effective control of sucking pests.

  16. Susceptibility profile of Aedes aegypti from Santiago Island, Cabo Verde, to insecticides.

    Science.gov (United States)

    Rocha, Hélio Daniel Ribeiro; Paiva, Marcelo Henrique Santos; Silva, Norma Machado; de Araújo, Ana Paula; Camacho, Denise Dos Reis da Rosa de Azevedo; Moura, Aires Januário Fernandes da; Gómez, Lara Ferrero; Ayres, Constância Flávia Junqueira; Santos, Maria Alice Varjal de Melo

    2015-12-01

    In 2009, Cabo Verde diagnosed the first dengue cases, with 21,137 cases reported and Aedes aegypti was identified as the vector. Since the outbreak, chemical insecticides and source reduction were used to control the mosquito population. This study aimed to assess the susceptibility of A. aegypti populations from Santiago, Cabo Verde to insecticides and identify the mechanisms of resistance. Samples of A. aegypti eggs were obtained at two different time periods (2012 and 2014), using ovitraps in different locations in Santiago Island to establish the parental population. F1 larvae were exposed to different concentrations of insecticides (Bacillus thuringiensis var israelensis (Bti), diflubenzuron and temephos) to estimate the lethal concentrations (LC90) and calculate the respective rate of resistance (RR90). Semi-field tests using temephos-ABATE(®) were performed to evaluate the persistence of the product. Bottle tests using female mosquitoes were carried out to determine the susceptibility to the adulticides malathion, cypermethrin and deltamethrin. Biochemical and molecular tests were performed to investigate the presence of metabolic resistance mechanisms, associated with the enzymes glutathione S-transferases (GSTs), esterases and mixed-function oxidases (MFO) and to detect mutations or alterations in the sodium channel and acetylcholinesterase genes. A. aegypti mosquitoes from Santiago exhibited resistance to deltamethrin, cypermethrin (mortality<80%) and temephos (RR90=4.4) but susceptibility to malathion (mortality≥98%), Bti and diflubenzuron. The low level of resistance to temephos did not affect the effectiveness of Abate(®). The enzymatic analysis conducted in 2012 revealed slight changes in the activities of GST (25%), MFO (18%), α-esterase (19%) and β-esterase (17%), but no significant changes in 2014. Target site resistance mutations were not detected. Our results suggest that the A. aegypti population from Santiago is resistant to two major

  17. Eficiência e Persistência de Três Produtos Comerciais à Base de Bacillus thuringiensis israelensis e Bacillus sphaericus no controle de Culicidae (Diptera em Lagoas de Tratamento de Efluentes

    Directory of Open Access Journals (Sweden)

    Jose Lopes

    2010-11-01

    Abstract. The hematofagic effect caused by females belonging to some species of Culicidae on humans and animals can be directly related to pathogen transmission, allergic reactions and uneasiness. The emergence of populations resistant to chemical insecticides has fostered the use of alternative methods, mainly biological control. The trials were conducted in three effluent treatment lagoons, on larvae of Culicidae to test the efficiency and persistence of commercial products whose active principles are based on Bacillus thuringiensis israelensis Berliner and Bacillus sphaericus Neide. The products tested were Vectolex (a granulated formulation of B. sphaericus, Sphaericus (a liquid formulation of B. sphaericus and Bt-horus (a liquid formulation of B. thuringiensis. The products were applied biweekly and evaluations were conducted 0, 24, 48, 72, and 120 hours after each application. The lagoons were colonized by Culex nigripalpus Theobald (1.5%, Culex saltanensis Dyar (2.25%, and Culex quinquefasciatus Say (96.25%. Bt-horus reduced larvae by 89.06%, 83.97% and 89.96% at 24, 48 and 72 hours after product application, respectively. The granulated and the liquid formulations containing B. sphaericus reduced larvae by 98.89 % and 98.34% 24 hours after application, and by 99.79% and 99.78% after 48 hours, respectively. The products and the different formulations were effective in controlling larvae of all three Culicidae species in lagoons with high levels of organic matter, but the persistence was recorded in two and three days for products containing respectively B. sphaericus and B. thuringiensis israelensis.

  18. EFEKTIVITAS Bacillus thuringiensis H-14 STRAIN LOKAL DALAM BUAH KELAPA TERHADAP LARVA Anopheles sp dan Culex sp di KAMPUNG LAUT KABUPATEN CILACAP

    Directory of Open Access Journals (Sweden)

    Blondine Ch. P

    2013-07-01

    Full Text Available Abstrak Bacillus thuringiensis serotipe H-14 strain lokal adalah bakteri patogen bersifat target spesifiknya larva nyamuk, aman bagi mamalia dan lingkungan. Penelitian bertujuan menentukan efektivitas B. thuringiensis H-14 strain lokal yang dikembangbiakkan dalam buah kelapa untuk pengendalian larva Anopheles sp dan Culex sp. Rancangan eksperimental semu, terdiri dari kelompok perlakuan dan kontrol. Bacillus thuringiensis H-14 strain lokal dikembangbiakan dalam10 buah kelapa umur 6–8 bulan, dengan berat kira-kira 1 kg, telah berisi air kelapa sekitar 400-500 ml/buah kelapa yang diperoleh dari Desa Klaces, Kampung Laut, Kabupaten Cilacap. Diinkubasi selama 14 hari pada temperatur kamar dan ditebarkan di 6 kolam yang menjadi habitat perkembangbiakan larva nyamuk dengan luas berkisar 3–100 m2.Hasil yang diperoleh menunjukkan efektivitas B. thuringiensis H-14 strain lokal terhadap larva Anopheles sp dan Culex sp selama 1 hari sesudah penebaran kematian larva berturut-turut sebesar 80–100% dan 79,31–100%. Sedangkan pada hari ke-14 sebesar 69,30–76,71% dan 67,69–86,04%. Buah kelapa dapat digunakan sebagai media lokal alternatif untuk pengembangbiakan B. thuringiensis H-14 strain lokal Kata kunci: B. thuringiensis H-14,  strain  lokal, buah kelapa, pengendalian larva Abstract Bacillus thuringiensis serotype H-14 local strain is pathogenic bacteria which specific  target to mosquito larvae. It is safe for mammals and enviroment. The aims of this study was to determine the effectivity of B. thuringiensis H-14 local strain which culturing in thecoconut wates against Anopheles sp and Culex sp mosquito larvae. This research is quasi experiment which consist of treated  and control groups. Bacillus thuringiensis H-14 local strain was cultured in 10 coconuts with 6–8 months age with weight around 1 kg that contained were approximately 400-500 ml/coconut were taken from Klaces village, Kampung Laut. After that the coconuts incubated for 14

  19. Transgenic loblolly pine (Pinus taeda L.) plants expressing a modified delta-endotoxin gene of Bacillus thuringiensis with enhanced resistance to Dendrolimus punctatus Walker and Crypyothelea formosicola Staud.

    Science.gov (United States)

    Tang, Wei; Tian, Yingchuan

    2003-02-01

    A synthetic version of the CRY1Ac gene of Bacillus thuringiensis has been used for the transformation of loblolly pine (Pinus taeda L.) using particle bombardment. Mature zygotic embryos were used to be bombarded and to generate organogenic callus and transgenic regenerated plants. Expression vector pB48.215 DNA contained a synthetic Bacillus thuringiensis (B.t.) CRY1Ac coding sequence flanked by the double cauliflower mosaic virus (CaMV) 35S promoter and nopaline synthase (NOS) terminator sequences, and the neomycin phosphotransferase II (NPTII) gene controlled by the promoter of the nopaline synthase gene was introduced into loblolly pine tissues by particle bombardment. The transformed tissues were proliferated and selected on media with kanamycin. Shoot regeneration was induced from the kanamycin-resistant calli, and transgenic plantlets were then produced. More than 60 transformed plants from independent transformation events were obtained for each loblolly pine genotype tested. The integration and expression of the introduced genes in the transgenic loblolly pine plants was confirmed by polymerase chain reactions (PCR) analysis, by Southern hybridization, by Northern blot analysis, and by Western blot analysis. Effective resistance of transgenic plants against Dendrolimus punctatus Walker and Crypyothelea formosicola Staud was verified in feeding bioassays with the insects. The transgenic plants recovered could represent a good opportunity to analyse the impact of genetic engineering of pine for sustainable resistance to pests using a B. thuringiensis insecticidal protein. This protocol enabled the routine transformation of loblolly pine plants that were previously difficult to transform.

  20. Multiple Cytochrome P450 genes: their constitutive overexpression and permethrin induction in insecticide resistant mosquitoes, Culex quinquefasciatus.

    Science.gov (United States)

    Liu, Nannan; Li, Ting; Reid, William R; Yang, Ting; Zhang, Lee

    2011-01-01

    Four cytochrome P450 cDNAs, CYP6AA7, CYP9J40, CYP9J34, and CYP9M10, were isolated from mosquitoes, Culex quinquefasciatus. The P450 gene expression and induction by permethrin were compared for three different mosquito populations bearing different resistance phenotypes, ranging from susceptible (S-Lab), through intermediate (HAmCq(G0), the field parental population) to highly resistant (HAmCq(G8), the 8(th) generation of permethrin selected offspring of HAmCq(G0)). A strong correlation was found for P450 gene expression with the levels of resistance and following permethrin selection at the larval stage of mosquitoes, with the highest expression levels identified in HAmCq(G8), suggesting the importance of CYP6AA7, CYP9J40, CYP9J34, and CYP9M10 in the permethrin resistance of larva mosquitoes. Only CYP6AA7 showed a significant overexpression in HAmCq(G8) adult mosquitoes. Other P450 genes had similar expression levels among the mosquito populations tested, suggesting different P450 genes may be involved in the response to insecticide pressure in different developmental stages. The expression of CYP6AA7, CYP9J34, and CYP9M10 was further induced by permethrin in resistant mosquitoes. Taken together, these results indicate that multiple P450 genes are up-regulated in insecticide resistant mosquitoes through both constitutive overexpression and induction mechanisms, thus increasing the overall expression levels of P450 genes.

  1. IFT20 modulates ciliary PDGFRα signaling by regulating the stability of Cbl E3 ubiquitin ligases

    DEFF Research Database (Denmark)

    Schmid, Fabian Marc; Schou, Kenneth Bødtker; Vilhelm, Martin Juel

    2018-01-01

    ciliogenesis, and ciliary localization of the receptor is required for its appropriate ligand-mediated activation by PDGF-AA. However, the mechanisms regulating sorting of PDGFRα and feedback inhibition of PDGFRα signaling at the cilium are unknown. Here, we provide evidence that intraflagellar transport...... protein 20 (IFT20) interacts with E3 ubiquitin ligases c-Cbl and Cbl-b and is required for Cbl-mediated ubiquitination and internalization of PDGFRα for feedback inhibition of receptor signaling. In wild-type cells treated with PDGF-AA, c-Cbl becomes enriched in the cilium, and the receptor...

  2. Structural basis for the recruitment and activation of the Legionella phospholipase VipD by the host GTPase Rab5

    Science.gov (United States)

    Lucas, María; Gaspar, Andrew H.; Pallara, Chiara; Rojas, Adriana Lucely; Fernández-Recio, Juan; Machner, Matthias P.; Hierro, Aitor

    2014-01-01

    A challenge for microbial pathogens is to assure that their translocated effector proteins target only the correct host cell compartment during infection. The Legionella pneumophila effector vacuolar protein sorting inhibitor protein D (VipD) localizes to early endosomal membranes and alters their lipid and protein composition, thereby protecting the pathogen from endosomal fusion. This process requires the phospholipase A1 (PLA1) activity of VipD that is triggered specifically on VipD binding to the host cell GTPase Rab5, a key regulator of endosomes. Here, we present the crystal structure of VipD in complex with constitutively active Rab5 and reveal the molecular mechanism underlying PLA1 activation. An active site-obstructing loop that originates from the C-terminal domain of VipD is repositioned on Rab5 binding, thereby exposing the catalytic pocket within the N-terminal PLA1 domain. Substitution of amino acid residues located within the VipD–Rab5 interface prevented Rab5 binding and PLA1 activation and caused a failure of VipD mutant proteins to target to Rab5-enriched endosomal structures within cells. Experimental and computational analyses confirmed an extended VipD-binding interface on Rab5, explaining why this L. pneumophila effector can compete with cellular ligands for Rab5 binding. Together, our data explain how the catalytic activity of a microbial effector can be precisely linked to its subcellular localization. PMID:25114243

  3. Dual function of a bee (Apis cerana) inhibitor cysteine knot peptide that acts as an antifungal peptide and insecticidal venom toxin.

    Science.gov (United States)

    Park, Hee Geun; Kyung, Seung Su; Lee, Kwang Sik; Kim, Bo Yeon; Choi, Yong Soo; Yoon, Hyung Joo; Kwon, Hyung Wook; Je, Yeon Ho; Jin, Byung Rae

    2014-12-01

    Inhibitor cysteine knot (ICK) peptides exhibit ion channel blocking, insecticidal, and antimicrobial activities, but currently, no functional roles for bee-derived ICK peptides have been identified. In this study, a bee (Apis cerana) ICK peptide (AcICK) that acts as an antifungal peptide and as an insecticidal venom toxin was identified. AcICK contains an ICK fold that is expressed in the epidermis, fat body, or venom gland and is present as a 6.6-kDa peptide in bee venom. Recombinant AcICK peptide (expressed in baculovirus-infected insect cells) bound directly to Beauveria bassiana and Fusarium graminearum, but not to Escherichia coli or Bacillus thuringiensis. Consistent with these findings, AcICK showed antifungal activity, indicating that AcICK acts as an antifungal peptide. Furthermore, AcICK expression is induced in the fat body and epidermis after injection with B. bassiana. These results provide insight into the role of AcICK during the innate immune response following fungal infection. Additionally, we show that AcICK has insecticidal activity. Our results demonstrate a functional role for AcICK in bees: AcICK acts as an antifungal peptide in innate immune reactions in the body and as an insecticidal toxin in venom. The finding that the AcICK peptide functions with different mechanisms of action in the body and in venom highlights the two-pronged strategy that is possible with the bee ICK peptide. Copyright © 2014 Elsevier Ltd. All rights reserved.

  4. An anionic defensin from Plutella xylostella with potential activity against Bacillus thuringiensis.

    Science.gov (United States)

    Xu, X-X; Zhang, Y-Q; Freed, S; Yu, J; Gao, Y-F; Wang, S; Ouyang, L-N; Ju, W-Y; Jin, F-L

    2016-12-01

    Insect defensins, are cationic peptides that play an important role in immunity against microbial infection. In the present study, an anionic defensin from Plutella xylostella, (designated as PxDef) was first cloned and characterized. Amino acid sequence analysis showed that the mature peptide owned characteristic six-cysteine motifs with predicted isoelectric point of 5.57, indicating an anionic defensin. Quantitative real-time polymerase chain reaction analysis showed that PxDef was significantly induced in epidermis, fat body, midgut and hemocytes after injection of heat-inactivated Bacillus thuringiensis, while such an induction was delayed by the injection of live B. thuringiensis in the 4th instar larvae of P. xylostella. Knocking down the expression of nuclear transcription factor Dorsal in P. xylostella by RNA interference significantly decreased the mRNA level of PxDef, and increased the sensitivity of P. xylostella larvae to the infection by live B. thuringiensis. The purified recombinant mature peptide (PxDef) showed higher activity against Gram-positive bacteria, with the minimum inhibition concentrations of 1.6 and 2.6 µM against B. thuringiensis and Bacillus subtilis, respectively. To our knowledge, this is the first report about an anionic PxDef, which may play an important role in the immune system of P. xylostella against B. thuringiensis.

  5. The effect of active immunization against vasoactive intestinal peptide (VIP) and inhibin on reproductive performance of aging White Leghorn roosters.

    Science.gov (United States)

    Avital-Cohen, N; Heiblum, R; Argov, N; Rosenstrauch, A; Chaiseha, Y; Mobarkey, N; Rozenboim, I

    2012-01-01

    Decreasing fertility in aging domestic roosters is a well-known phenomenon. Aging is manifested by a decrease in plasma testosterone level, testis function, and spermatogenesis, resulting in a low level of fertility. The roles of vasoactive intestinal peptide (VIP) and testicular inhibin in this aging process are not clear. The effects of active immunization against VIP, inhibin, or the combination of both hormones on the reproduction of aging White Leghorn (WL) roosters were assayed. In experiment 1a, 60 White Leghorn roosters (67 wk of age) were divided into 4 groups (n = 15/group). The first group was actively immunized against VIP, the second against inhibin, the third against VIP and inhibin, and the fourth served as a control. Active immunization against VIP decreased semen quality parameters, plasma steroid levels, and gene expression of gonadotropin-releasing hormone-I (GnRH-I), follicle-stimulating hormone (FSH), luteinizing hormone (LH), LH receptor, VIP, and prolactin (Prl). Immunization against inhibin increased some of the semen quality parameters and FSH mRNA gene expression but decreased inhibin gene expression. In experiment 1b, at 94 wk of age, we took the actively immunized against VIP group and the control group and divided them into 2 subgroups (n = 7 or 8): the first group was injected with 1 mg of ovine Prl (oPrl) daily for 7 d, and the second group served as a control. Administration of oPrl to previously VIP-immunized birds significantly elevated semen quality parameters. We suggest that VIP, Prl, and inhibin have an important effect on the reproductive axis in aging roosters. Active immunization against VIP-depressed reproductive activity and Prl administration restored their reproduction, indicating that both VIP and Prl are essential for reproduction in aging roosters. Immunization against inhibin improved FSH mRNA gene expression, suggesting a negative role of inhibin on FSH secretion in aging roosters. Not all semen quality parameters

  6. First Lady Hillary Clinton waits for launch at VIP viewing site

    Science.gov (United States)

    1999-01-01

    Among those gathered at the VIP viewing site for the launch of STS-93 are First Lady Hillary Rodham Clinton (center) and Donna Shalala, secretary , Department of Health and Human Services (far right). Also present is Chelsea Clinton. Much attention has been generated over the launch due to Commander Eileen M. Collins, the first woman to serve as commander of a Shuttle mission. The primary payload of the five-day mission is the Chandra X-ray Observatory, which will allow scientists from around the world to study some of the most distant, powerful and dynamic objects in the universe. The new telescope is 20 to 50 times more sensitive than any previous X-ray telescope and is expected to unlock the secrets of supernovae, quasars and black holes. Liftoff of Space Shuttle Columbia is scheduled for 12:36 a.m. EDT July 20.

  7. mGluR1 receptors contribute to non-purinergic slow excitatory transmission to submucosal VIP neurons of guinea-pig ileum

    Directory of Open Access Journals (Sweden)

    Jaime Pei Pei Foong

    2009-05-01

    Full Text Available Vasoactive intestinal peptide (VIP immunoreactive secretomotor neurons in the submucous plexus are involved in mediating bacterial toxin-induced hypersecretion leading to diarrhoea. VIP neurons become hyperexcitable after the mucosa is exposed to cholera toxin, which suggests that the manipulation of the excitability of these neurons may be therapeutic. This study used standard intracellular recording methods to systematically characterize slow excitatory postsynaptic potentials (EPSPs evoked in submucosal VIP neurons by different stimulus regimes (1, 3 and 15 pulse 30 Hz stimulation, together with their associated input resistances and pharmacology. All slow EPSPs were associated with a significant increase in input resistance compared to baseline values. Slow EPSPs evoked by a single stimulus were confirmed to be purinergic, however, slow EPSPs evoked by 15 pulse trains were non-purinergic and those evoked by 3 pulse trains were mixed. NK1 or NK3 receptor antagonists did not affect slow EPSPs. The group I mGluR receptor antagonist, PHCCC reduced the amplitude of purinergic and non-purinergic slow EPSPs. Blocking mGluR1 receptors depressed the overall response to 3 and 15 pulse trains, but this effect was inconsistent, while blockade of mGluR5 receptors had no effect on the non-purinergic slow EPSPs. Thus, although other receptors are almost certainly involved, our data indicate that there are at least two pharmacologically distinct types of slow EPSPs in the VIP secretomotor neurons: one mediated by P2Y receptors and the other in part by mGluR1 receptors.

  8. Evaluación de dos formulaciones de Bacillus thuringiensis H-14 para el control de larvas de Aedes aegypti

    Directory of Open Access Journals (Sweden)

    Marco F. Suárez

    1987-06-01

    Full Text Available Se evaluaron en el laboratorio y en condiciones naturales, dos formulaciones de Bacillus thuringiensis H-14, el cual es un insecticida microbiológico contra larvas de Aedes aegypti Se utilizó polvo humectable (3.500 AA/mg y líquido concentrado (1.000 AA/mg (AA = Unidades Tóxicas Internacionales en A. aegypti. Se calcularon las dosis letales (DL 99,9 para cada formulación. En una estación de laboratorio que simulaba las condiciones naturales de cría de A. aegypti, se realizaron pruebas con cuatro dosis diferentes de cada una de las dos formulaciones. Con una dosis de 1.050 AA/I, se encontró una acción residual de 11 días con el polvo humectable y de 9,9 días con el líquido concentrado. La acción residual no aumentó en proporción directa con la dosis. La tasa de emergencia diaria de adultos tardó 21,7 días en recuperarse hasta el 30% de su nivel de pretratamiento, cuando se aplicó polvo humectable y 18 días cuando se aplicó Iíquido concentrado. Aunque en las pruebas en condiciones seminaturales, se encontraron diferencias estadísticamente significativas entre las diferentes dosis y los diferentes tipos de criaderos, operacionalmente estas diferencias no tienen importancia práctica porque no aumenta el rendimiento de la medida de control. Los ensayos en condiciones naturales se realizaron aplicando una dosis de 1.050 AA/I a las albercas, toneles y llantas. El B. t. H-14 demostró ser efectivo, al causar 100% de mortalidad en todos los estadios larvarios de A. aegyptien los criaderos tratados. Se obtuvo una acción residual de 9 días en los criaderos cuando se aplicó polvo humectable y 7,2 días cuando se aplicó líquido concentrado. El uso del B. t. H-14. como larvicida alterno, tiene posibilidades en situaciones especiales, donde los métodos químicosson indeseadoso imprácticos.

  9. Effective dominance of resistance of Spodoptera frugiperda to Bt maize and cotton varieties: implications for resistance management

    Science.gov (United States)

    Horikoshi, Renato J.; Bernardi, Daniel; Bernardi, Oderlei; Malaquias, José B.; Okuma, Daniela M.; Miraldo, Leonardo L.; Amaral, Fernando S. De A. E.; Omoto, Celso

    2016-10-01

    The resistance of fall armyworm (FAW), Spodoptera frugiperda, has been characterized to some Cry and Vip3A proteins of Bacillus thuringiensis (Bt) expressed in transgenic maize in Brazil. Here we evaluated the effective dominance of resistance based on the survival of neonates from selected Bt-resistant, heterozygous, and susceptible (Sus) strains of FAW on different Bt maize and cotton varieties. High survival of strains resistant to the Cry1F (HX-R), Cry1A.105/Cry2Ab (VT-R) and Cry1A.105/Cry2Ab/Cry1F (PW-R) proteins was detected on Herculex, YieldGard VT PRO and PowerCore maize. Our Vip3A-resistant strain (Vip-R) exhibited high survival on Herculex, Agrisure Viptera and Agrisure Viptera 3 maize. However, the heterozygous from HX-R × Sus, VT-R × Sus, PW-R × Sus and Vip-R × Sus had complete mortality on YieldGard VT PRO, PowerCore, Agrisure Viptera, and Agrisure Viptera 3, whereas the HX-R × Sus and Vip-R × Sus strains survived on Herculex maize. On Bt cotton, the HX-R, VT-R and PW-R strains exhibited high survival on Bollgard II. All resistant strains survived on WideStrike, but only PW-R and Vip-R × Sus survived on TwinLink. Our study provides useful data to aid in the understanding of the effectiveness of the refuge strategy for Insect Resistance Management of Bt plants.

  10. Bioassay method for toxicity studies of insecticide formulations to Tuta absoluta (meyrick, 1917 Metodologia de bioensaio para estudos de toxicidade de formulações comerciais de inseticidas a Tuta absoluta (Meyrick, 1917

    Directory of Open Access Journals (Sweden)

    Tarcísio Visintin da Silva Galdino

    2011-10-01

    Full Text Available Chemical control is the main method for controlling the tomato leafminer, Tuta absoluta (Meyrick, 1917 (Lepidoptera: Gelechiidae. Reported techniques for the evaluation of insecticide toxicity to the tomato leafminer are not in agreement with field conditions and do not allow us to verify whether doses used in the field are efficient for control. Thus, the objective of this work was to develop a bioassay methodology to study the toxicity of insecticide formulations to T. absoluta that represent field conditions for fast-acting insecticides (neurotoxics and inhibitors of respiration and slow-acting insecticides (Bacillus thuringiensis and insect growth regulators. The leaf-dip method was the most efficient method for toxicity studies of insecticides formulations to T. absoluta. We verified that bioassays with fast-acting insecticides should be performed with glass Petri dishes containing one tomato foliole from the 4th leaf from the plant apex infested with 10 larvae of 3rd instar and these bioassays can last 48 hours. Conversely, bioassays with slow-acting insecticides should be performed with two-liter transparent PET bottles containing the 4th leaf from the plant apex, with their petioles immersed in a glass bottle containing 120 mL of water, and this leaf should be infested with 10 larvae of 2nd instar and this bioassays can last seven days.O principal método utilizado no controle da traça-do-tomateiro Tuta absoluta (Meyrick, 1917 (Lepidoptera: Gelechiidae é a aplicação de inseticidas. As técnicas atuais de avaliação da toxicidade de inseticidas sobre essa praga não simulam a situação de campo e não possibilitam a verificação se as doses usadas no campo são eficientes no seu controle. Assim, neste trabalho, objetivou-se desenvolver uma metodologia que represente as condições de campo para inseticidas de ação rápida (neurotóxicos e inibidores respiratórios e de ação lenta (Bacillus thuringiensis e reguladores de

  11. An Intramolecular Salt Bridge in Bacillus thuringiensis Cry4Ba Toxin Is Involved in the Stability of Helix α-3, Which Is Needed for Oligomerization and Insecticidal Activity.

    Science.gov (United States)

    Pacheco, Sabino; Gómez, Isabel; Sánchez, Jorge; García-Gómez, Blanca-Ines; Soberón, Mario; Bravo, Alejandra

    2017-10-15

    Bacillus thuringiensis three-domain Cry toxins kill insects by forming pores in the apical membrane of larval midgut cells. Oligomerization of the toxin is an important step for pore formation. Domain I helix α-3 participates in toxin oligomerization. Here we identify an intramolecular salt bridge within helix α-3 of Cry4Ba (D111-K115) that is conserved in many members of the family of three-domain Cry toxins. Single point mutations such as D111K or K115D resulted in proteins severely affected in toxicity. These mutants were also altered in oligomerization, and the mutant K115D was more sensitive to protease digestion. The double point mutant with reversed charges, D111K-K115D, recovered both oligomerization and toxicity, suggesting that this salt bridge is highly important for conservation of the structure of helix α-3 and necessary to promote the correct oligomerization of the toxin. IMPORTANCE Domain I has been shown to be involved in oligomerization through helix α-3 in different Cry toxins, and mutations affecting oligomerization also elicit changes in toxicity. The three-dimensional structure of the Cry4Ba toxin reveals an intramolecular salt bridge in helix α-3 of domain I. Mutations that disrupt this salt bridge resulted in changes in Cry4Ba oligomerization and toxicity, while a double point reciprocal mutation that restored the salt bridge resulted in recovery of toxin oligomerization and toxicity. These data highlight the role of oligomer formation as a key step in Cry4Ba toxicity. Copyright © 2017 American Society for Microbiology.

  12. Isolation and characterization of a novel native Bacillus thuringiensis strain BRC-HZM2 capable of degrading chlorpyrifos.

    Science.gov (United States)

    Wu, Songqing; Peng, Yan; Huang, Zhangmin; Huang, Zhipeng; Xu, Lei; Ivan, Gelbič; Guan, Xiong; Zhang, Lingling; Zou, Shuangquan

    2015-03-01

    Studies were carried out to isolate chlorpyrifos degrading Bacillus thuringiensis (Bt) strains from chlorpyrifos-contaminated samples. Six Bt strains (isolation rate 2.7%) were isolated by modified sodium acetate antibiotic heat treatment, and one novel strain (BRC-HZM2) was selected for further analysis. Phenotype and phylogeny analysis of this strain was conducted on the basis of biochemical reactions, antibiotic sensitivity, 16s rRNA genes, plasmid profile, insecticidal crystal protein profiles, and PCR-RFLP for cry and cyt genes. The degradation rate of chlorpyrifos in liquid culture was estimated during 48 h of incubation for the isolate BRC-HZM2. More than 50% of the initial chlorpyrifos concentration degraded within 12 h, 88.9% after 48 h. These results highlight the potential of the Bt strain for biological control and the bioremediation of environments contaminated with chlorpyrifos. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  13. Chitinase production by Bacillus thuringiensis and Bacillus licheniformis: their potential in antifungal biocontrol.

    Science.gov (United States)

    Gomaa, Eman Zakaria

    2012-02-01

    Thirty bacterial strains were isolated from the rhizosphere of plants collected from Egypt and screened for production of chitinase enzymes. Bacillus thuringiensis NM101-19 and Bacillus licheniformis NM120-17 had the highest chitinolytic activities amongst those investigated. The production of chitinase by B. thuringiensis and B. licheniformis was optimized using colloidal chitin medium amended with 1.5% colloidal chitin, with casein as a nitrogen source, at 30°C after five days of incubation. An enhancement of chitinase production by the two species was observed by addition of sugar substances and dried fungal mats to the colloidal chitin media. The optimal conditions for chitinase activity by B. thuringiensis and B. licheniformis were at 40°C, pH 7.0 and pH 8.0, respectively. Na(+), Mg(2+), Cu(2+), and Ca(2+) caused enhancement of enzyme activities whereas they were markedly inhibited by Zn(2+), Hg(2+), and Ag(+). In vitro, B. thuringiensis and B. licheniformis chitinases had potential for cell wall lysis of many phytopathogenic fungi tested. The addition of B. thuringiensis chitinase was more effective than that of B. licheniformis in increasing the germination of soybean seeds infected with various phytopathogenic fungi.

  14. Vasoactive intestinal polypeptide (VIP) in cirrhosis: arteriovenous extraction in different vascular beds

    DEFF Research Database (Denmark)

    Henriksen, Jens Henrik Sahl; Staun-Olsen, P; Fahrenkrug, J

    1980-01-01

    The concentration of vasoactive intestinal polypeptide (VIP) was determined in peripheral venous plasma from 136 patients with liver cirrhosis without gastrointestinal bleeding or coma and from 112 controls. In eight patients (cirrhosis, six; fibrosis, one; steatosis, one) arteriovenous extraction...... is significantly elevated in peripheral plasma from patients with cirrhosis, probably due to porto-systemic shunting and/or compromised hepatic elimination. Hepatic elimination is still likely to account for the inactivation of most of the VIP escaping from the neurosynapses throughout the body in patients...

  15. [Exposure degree of important non-target arthropods to Cry2Aa in Bt rice fields].

    Science.gov (United States)

    Zhang, Qing-Ling; Li, Yun-He; Hua, Hong-Xia; Yang, Chang-Ju; Wu, Hong-Jin; Peng, Yu-Fa

    2013-06-01

    Based on the principle of "risk = hazard x exposure", the selected representative nontarget organisms in the assessment of the potential effects of insect-resistant genetically modified (GM) crops on non-target arthropods in laboratory are generally the arthropod species highly exposed to the insecticidal proteins expressed by the GM crops in farmland ecosystem. In order to understand the exposure degree of the important arthropod species to Cry proteins in Bt rice fields, and to select the appropriate non-target arthropods in the risk assessment of insect-resistant GM crops, the enzyme-linked immunosorbent assay (ELISA) was conducted to measure the Cry2Aa protein concentration in the arthropods collected from the cry2Aa rice fields at different rice growth stages. The results showed that there was a significant difference in the Cry2Aa content protein concentration in different arthropod species. Some species did not contain Cry2Aa protein, while some species contained larger amounts of Cry2Aa protein. Relative to the arthropods colleted after rice anthesis, the arthropods colleted in rice anthesis contained relative higher concentrations of Cry2Aa protein, especially for the predacious arthropods. No Cry proteins were detected in parasitic arthropods. This study provided references for the laboratory assessment of the effects of GM rice on nontarget arthropods.

  16. Characterization of cry1Cb3 and cry1Fb7 from Bacillus thuringiensis subsp. galleriae

    Czech Academy of Sciences Publication Activity Database

    Huang, T.; Xiao, Y.; Pan, J.; Zhang, L.; Gelbič, Ivan; Guan, X.

    2015-01-01

    Roč. 10, č. 1 (2015), s. 521-528 ISSN 2391-5412 Institutional support: RVO:60077344 Keywords : Bacillus thuringiensis subsp. galleriae * PCR-RFLP * cloning Subject RIV: EB - Genetics ; Molecular Biology http://www.degruyter.com/view/j/biol.2015.10.issue-1/biol-2015-0054/biol-2015-0054.xml

  17. [The expression and significance of VIP and its receptor in the cochlea of different degrees of chronic alcoholism rats].

    Science.gov (United States)

    Feng, Jing; Liu, Haibing

    2015-07-01

    To determine whether chronic alcoholism alters the expression levels of Vasoactive intestinal polypeptide (VIP) and its receptor (VIPR1) in the cochlea of chronic alcoholism rats. We measured their expression levels in 30 SD rats, in which we created models of different degrees of chronic alcoholism. We investigated the presence of the mRNA of VIP in the cochlea of chronic alcoholism rats and controls by reverse transcription-polymerase chain reaction (RT-PCR) method. We investigated the presence of proteins of VIPR1 in poisoned rats and controls by western blot. We also evaluated the local distribution of VIP cells by immunohistochemistry. We found that the levels of VIP and VIPR1 were downregulated in the chronic alcoholism groups compared to the controls group. The differences in some expression levels were significant different between chronic alcoholism rats and control rats. Moreover, at different degrees of alcohol poisoning in rats, the contents of VIP and VIPR1 differed. Decreased levels of VIP and VIPR1 were detected in the deep chronic alcoholism group compared to the group with low-degree poisoning (P 0.05). These results suggest that VIP and VIPR1 play an important role in the auditory function in rats with chronic alcoholism. Chronic alcoholism may cause a peptide hormone secretion imbalance in the auditory system, eventually leading to hearing loss.

  18. Atypical nuclear localization of VIP receptors in glioma cell lines and patients

    Energy Technology Data Exchange (ETDEWEB)

    Barbarin, Alice; Séité, Paule [Equipe Récepteurs, Régulations et Cellules Tumorales, Université de Poitiers, PBS bât 36, 1 rue Georges Bonnet, TSA 51106, 86073 Poitiers Cedex 9 (France); Godet, Julie [Laboratoire d’anatomie et de cytologie pathologiques, CHU de Poitiers, 2 rue de la Milétrie, 86000 Poitiers (France); Bensalma, Souheyla; Muller, Jean-Marc [Equipe Récepteurs, Régulations et Cellules Tumorales, Université de Poitiers, PBS bât 36, 1 rue Georges Bonnet, TSA 51106, 86073 Poitiers Cedex 9 (France); Chadéneau, Corinne, E-mail: corinne.chadeneau@univ-poitiers.fr [Equipe Récepteurs, Régulations et Cellules Tumorales, Université de Poitiers, PBS bât 36, 1 rue Georges Bonnet, TSA 51106, 86073 Poitiers Cedex 9 (France)

    2014-11-28

    Highlights: • The VIP receptor VPAC1 contains a putative NLS signal. • VPAC1 is predominantly nuclear in GBM cell lines but not VPAC2. • Non-nuclear VPAC1/2 protein expression is correlated with glioma grade. • Nuclear VPAC1 is observed in 50% of stage IV glioma (GBM). - Abstract: An increasing number of G protein-coupled receptors, like receptors for vasoactive intestinal peptide (VIP), are found in cell nucleus. As VIP receptors are involved in the regulation of glioma cell proliferation and migration, we investigated the expression and the nuclear localization of the VIP receptors VPAC1 and VPAC2 in this cancer. First, by applying Western blot and immunofluorescence detection in three human glioblastoma (GBM) cell lines, we observed a strong nuclear staining for the VPAC1 receptor and a weak nuclear VPAC2 receptor staining. Second, immunohistochemical staining of VPAC1 and VPAC2 on tissue microarrays (TMA) showed that the two receptors were expressed in normal brain and glioma tissues. Expression in the non-nuclear compartment of the two receptors significantly increased with the grade of the tumors. Analysis of nuclear staining revealed a significant increase of VPAC1 staining with glioma grade, with up to 50% of GBM displaying strong VPAC1 nuclear staining, whereas nuclear VPAC2 staining remained marginal. The increase in VPAC receptor expression with glioma grades and the enhanced nuclear localization of the VPAC1 receptors in GBM might be of importance for glioma progression.

  19. Expression of a Chimeric Gene Encoding Insecticidal Crystal Protein Cry1Aabc of Bacillus thuringiensis in Chickpea (Cicer arietinum L.) Confers Resistance to Gram Pod Borer (Helicoverpa armigera Hubner.).

    Science.gov (United States)

    Das, Alok; Datta, Subhojit; Thakur, Shallu; Shukla, Alok; Ansari, Jamal; Sujayanand, G K; Chaturvedi, Sushil K; Kumar, P A; Singh, N P

    2017-01-01

    Domain swapping and generation of chimeric insecticidal crystal protein is an emerging area of insect pest management. The lepidopteran insect pest, gram pod borer ( Helicoverpa armigera H.) wreaks havoc to chickpea crop affecting production. Lepidopteran insects were reported to be controlled by Bt ( cryI ) genes. We designed a plant codon optimized chimeric Bt gene ( cry1Aabc ) using three domains from three different cry1A genes (domains I, II, and III from cry1Aa , cry1Ab , and cry1Ac , respectively) and expressed it under the control of a constitutive promoter in chickpea ( cv . DCP92-3) to assess its effect on gram pod borer. A total of six transgenic chickpea shoots were established by grafting into mature fertile plants. The in vitro regenerated (organogenetic) shoots were selected based on antibiotic kanamycin monosulfate (100 mg/L) with transformation efficiency of 0.076%. Three transgenic events were extensively studied based on gene expression pattern and insect mortality across generations. Protein expression in pod walls, immature seeds and leaves (pre- and post-flowering) were estimated and expression in pre-flowering stage was found higher than that of post-flowering. Analysis for the stable integration, expression and insect mortality (detached leaf and whole plant bioassay) led to identification of efficacious transgenic chickpea lines. The chimeric cry1Aabc expressed in chickpea is effective against gram pod borer and generated events can be utilized in transgenic breeding program.

  20. Expression of a Chimeric Gene Encoding Insecticidal Crystal Protein Cry1Aabc of Bacillus thuringiensis in Chickpea (Cicer arietinum L. Confers Resistance to Gram Pod Borer (Helicoverpa armigera Hubner.

    Directory of Open Access Journals (Sweden)

    Alok Das

    2017-08-01

    Full Text Available Domain swapping and generation of chimeric insecticidal crystal protein is an emerging area of insect pest management. The lepidopteran insect pest, gram pod borer (Helicoverpa armigera H. wreaks havoc to chickpea crop affecting production. Lepidopteran insects were reported to be controlled by Bt (cryI genes. We designed a plant codon optimized chimeric Bt gene (cry1Aabc using three domains from three different cry1A genes (domains I, II, and III from cry1Aa, cry1Ab, and cry1Ac, respectively and expressed it under the control of a constitutive promoter in chickpea (cv. DCP92-3 to assess its effect on gram pod borer. A total of six transgenic chickpea shoots were established by grafting into mature fertile plants. The in vitro regenerated (organogenetic shoots were selected based on antibiotic kanamycin monosulfate (100 mg/L with transformation efficiency of 0.076%. Three transgenic events were extensively studied based on gene expression pattern and insect mortality across generations. Protein expression in pod walls, immature seeds and leaves (pre- and post-flowering were estimated and expression in pre-flowering stage was found higher than that of post-flowering. Analysis for the stable integration, expression and insect mortality (detached leaf and whole plant bioassay led to identification of efficacious transgenic chickpea lines. The chimeric cry1Aabc expressed in chickpea is effective against gram pod borer and generated events can be utilized in transgenic breeding program.

  1. In vitro ovicidal and cestocidal effects of toxins from Bacillus thuringiensis on the canine and human parasite Dipylidium caninum.

    Science.gov (United States)

    Peña, Guadalupe; Aguilar Jiménez, Fortino Agustín; Hallal-Calleros, Claudia; Morales-Montor, Jorge; Hernández-Velázquez, Víctor Manuel; Flores-Pérez, Fernando Iván

    2013-01-01

    Bacillus thuringiensis is a gram-positive soil-dwelling bacterium that is commonly used as a biological pesticide. This bacterium may also be used for biological control of helminth parasites in domestic animals. In this study, we evaluated the possible ovicidal and cestocidal effects of a total protein extract of B. thuringiensis native strains on the zoonotic cestode parasite of dogs, Dipylidium caninum (D. caninum). Dose and time response curves were determined by coincubating B. thuringiensis proteins at concentration ranging from 100 to 1000 μ g/mL along with 4000 egg capsules of D. caninum. Egg viability was evaluated using the trypan blue exclusion test. The lethal concentration of toxins on eggs was 600 μ g/ml, and the best incubation time to produce this effect was 3 h. In the adult stage, the motility and the thickness of the tegument were used as indicators of damage. The motility was inhibited by 100% after 8 hours of culture compared to the control group, while the thickness of the cestode was reduced by 34%. Conclusively, proteins of the strain GP526 of B. thuringiensis directly act upon D. caninum showing ovicidal and cestocidal effects. Thus, B. thuringiensis is proposed as a potential biological control agent against this zoonosis.

  2. In Vitro Ovicidal and Cestocidal Effects of Toxins from Bacillus thuringiensis on the Canine and Human Parasite Dipylidium caninum

    Directory of Open Access Journals (Sweden)

    Guadalupe Peña

    2013-01-01

    Full Text Available Bacillus thuringiensis is a gram-positive soil-dwelling bacterium that is commonly used as a biological pesticide. This bacterium may also be used for biological control of helminth parasites in domestic animals. In this study, we evaluated the possible ovicidal and cestocidal effects of a total protein extract of B. thuringiensis native strains on the zoonotic cestode parasite of dogs, Dipylidium caninum (D. caninum. Dose and time response curves were determined by coincubating B. thuringiensis proteins at concentration ranging from 100 to 1000 μg/mL along with 4000 egg capsules of D. caninum. Egg viability was evaluated using the trypan blue exclusion test. The lethal concentration of toxins on eggs was 600 μg/ml, and the best incubation time to produce this effect was 3 h. In the adult stage, the motility and the thickness of the tegument were used as indicators of damage. The motility was inhibited by 100% after 8 hours of culture compared to the control group, while the thickness of the cestode was reduced by 34%. Conclusively, proteins of the strain GP526 of B. thuringiensis directly act upon D. caninum showing ovicidal and cestocidal effects. Thus, B. thuringiensis is proposed as a potential biological control agent against this zoonosis.

  3. Characterization of UDP-glucuronosyltransferase genes and their possible roles in multi-insecticide resistance in Plutella xylostella (L.).

    Science.gov (United States)

    Li, Xiuxia; Shi, Haiyan; Gao, Xiwu; Liang, Pei

    2018-03-01

    Uridine diphosphate-glucuronosyltransferases (UGTs), as multifunctional detoxification enzymes, play important roles in the biotransformation of various compounds. However, their roles in insecticide resistance are still unclear. This study presents a genome-wide identification of the UGTs in diamondback moth, Plutella xylostella (L.), a notorious insect pest of cruciferous crops worldwide. The possible roles of these UGTs in insecticide resistance were evaluated. A total of 21 putative UGTs in P. xylostella were identified. Quantitative real-time polymerase chain reaction (PCR)-based analyses showed that all the UGT genes were expressed in all tested developmental stages and tissues. Bioassay results indicated that a field-collected population (BL) was resistant to 9 of 10 commonly used insecticides, and 10 of 21 UGT mRNAs were upregulated in the BL population. Exposure to the LC 50 of each insecticide affected the expression of most UGT genes. Among these, the expression levels of UGT40V1, UGT45B1 and UGT33AA4 were induced by more than five insecticides, whereas indoxacarb and metaflumizone significantly repressed the expression of most UGT genes. UGTs may play important roles in the metabolism of commonly used insecticides in P. xylostella. These findings provide valuable information for further research on the physiological and toxicological functions of specific UGT genes in P. xylostella. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

  4. Insecticide solvents: interference with insecticidal action.

    Science.gov (United States)

    Brattsten, L B; Wilkinson, C F

    1977-06-10

    Several commercial solvent mixtures commonly used as insecticide carriers in spray formulations increase by more than threefold the microsomal N-demethylation of p-chloro N-methylaniline in midgut preparations of southern army-worm (Spodoptera eridania) larvae exposed orally to the test solvents. Under laboratory conditions, the same solvent mixtures exhibit a protective action against the in vivo toxicity of the insecticide carbaryl to the larvae. The data are discussed with respect to possible solvent-insecticide interactions occurring under field conditions and, more broadly, to potential toxicological hazards of these solvents to humans.

  5. 99mTc labeled VIP analog: evaluation for imaging colorectal cancer

    International Nuclear Information System (INIS)

    Rao, P.S.; Thakur, M.L.; Pallela, V.; Patti, R.; Reddy, K.; Li, H.; Sharma, S.; Pham, H.L.; Diggles, L.; Minami, C.; Marcus, C.S.

    2001-01-01

    Early and reliable diagnosis of colorectal cancer continues to be demanding and challenging. Colorectal cancer cells express Vasoactive Intestinal Peptide (VIP) receptors in high density. We have prepared a VIP analog (TP3654), labeled it with 99m Tc, and evaluated it in experimental animals as an agent for imaging colorectal cancer. The tissue distribution of 99m Tc-TP3654 has been compared with that of 111 In-DTPA-Octreotide and 99m Tc-anti-CEA scan in nude mice bearing human colorectal cancer LS174T. Finally, pharmacokinetic and tissue distribution studies of 99m Tc-TP3654 have been performed in four normal human volunteers. Data suggest that 99m Tc-TP3654 can be prepared efficiently without loss of its receptor specificity and biological activity. Although the 24 hr tumor uptake of 99m Tc-TP3654 in the animal model used was modest (0.21 ± 0.07% I.D./g), the tissue distribution profile was more favorable than that of 111 In-DTPA-Octreotide or 99m Tc-anti-CEA scan. Human studies indicated that 99m Tc-TP3654 had no adverse effect in any subject. Within 24 hours, approximately 70% of the injected dose cleared through the kidneys, and approximately 20% through the hepatobiliary system. In these non-fasting volunteers hepatobiliary clearance was slow and in cancer patients tumor uptake was rapid. Data suggest that 99m Tc-TP3654 is a promising agent for imaging colorectal cancer

  6. In vitro effect of Bacillus thuringiensis strains and Cry proteins in phytopathogenic fungi of paddy rice-field Efeito in vitro de cepas e proteínas Cry de Bacillus thuringiensis em fungos fitopatogênicos da cultura do arroz irrigado

    Directory of Open Access Journals (Sweden)

    Neiva Knaak

    2007-09-01

    Full Text Available Cry1Ab and Cry1Ac strains and proteins synthesized by Bacillus thuringiensis thuringiensis and B. thuringiensis kurstaki were assessed in the following phytopathogens: Rhizoctonia solani,Pyricularia grisea,Fusarium oxysporum and F. solani, which had their micelial growth decreased after incubation in the presence of the bacterial strains. As to Cry proteins, there were no inhibition halo development in the assessed concentrations.As cepas e proteínas Cry1Ab e Cry1Ac sintetizadas por Bacillus thuringiensis thuringiensis e B. thuringiensis kurstaki, foram avaliadas nos fitopatógenos: Rhizoctonia solani,Pyricularia grisea,Fusarium oxysporum e F. solani, os quais tiveram seu crescimento micelial reduzido após a incubação na presença das cepas bacterianas. Em relação às proteínas Cry, não houve formação de halo de inibição nas concentrações avaliadas.

  7. Decolorization of textile dye RB19 using volcanic rock matrix immobilized Bacillus thuringiensis cells with surface displayed laccase.

    Science.gov (United States)

    Wan, Juan; Sun, Xiaowen; Liu, Cheng; Tang, Mengjun; Li, Lin; Ni, Hong

    2017-06-01

    A triplicate volcanic rock matrix-Bacillus thuringiensis-laccase WlacD (VRMs-Bt-WlacD) dye decolorization system was developed. WlacD was displayed on the B. thuringiensis MB174 cell surface to prepare a whole-cell laccase biocatalyst by using two repeat N-terminal domains of autolysin Mbg (Mbgn) 2 as the anchoring motif. Immunofluorescence microscopic assays confirmed that the fusion protein (Mbgn) 2 -WlacD was anchored on the surface of the recombinant B. thuringiensis MB174. After optimization by a single factor test, L 9 (3 4 )-orthogonal test, Plackett-Burman test, steepest ascent method, and Box-Behnken response surface methodology, the whole-cell specific laccase activity of B. thuringiensis MB174 was improved to 555.2 U L -1 , which was 2.25 times than that of the primary culture condition. Optimized B. thuringiensis MB174 cells were further adsorbed by VRMs to prepare VRMs-Bt-WlacD, an immobilized whole-cell laccase biocatalyst. Decolorization capacity of as-prepared VRMs-Bt-WlacD toward an initial concentration of 500 mg L -1 of an textile dye reactive blue 19 (RB19) aqueous solution reached 72.36% at a solid-to-liquid ratio of 10 g-100 mL. Repeated decolorization-activation operations showed the high decolorization capacity of VRMs-Bt-WlacD and have the potential for large-scale or continuous operations.

  8. Pengaruh Rapat Arus Anodizing terhadap Nilai Kekerasan pada Plat Aluminium Paduan Aa Seri 2024-t3

    OpenAIRE

    Fajar Nugroho

    2015-01-01

    Aluminum alloy AA 2024-T3 is widely applied in the aircraft industry because it has good mechanical properties such as; light weight, good conductivity and the corrosion resistance. However Aluminium 2024-T3 susceptible to wearing. One method to improve the wear resistance o f AA 2024-T3 is the anodizing process. The aims of this research to study the effect of current density and anodizing time against the hardness of aluminum alloy AA 2024-T3. The process of anodizing was carried out using ...

  9. Crystallization and preliminary crystallographic analysis of poly(3-hydroxybutyrate) depolymerase from Bacillus thuringiensis.

    Science.gov (United States)

    Wang, Yung Lin; Lin, Yi Ting; Chen, Chia Lin; Shaw, Gwo Chyuan; Liaw, Shwu Huey

    2014-10-01

    Poly[(R)-3-hydroxybutyrate] (PHB) is a microbial biopolymer that has been commercialized as biodegradable plastics. The key enzyme for the degradation is PHB depolymerase (PhaZ). A new intracellular PhaZ from Bacillus thuringiensis (BtPhaZ) has been screened for potential applications in polymer biodegradation. Recombinant BtPhaZ was crystallized using 25% polyethylene glycol 3350, 0.2 M ammonium acetate, 0.1 M bis-tris pH 6.5 at 288 K. The crystals belonged to space group P1, with unit-cell parameters a = 42.97, b = 83.23, c = 85.50 Å, α = 73.45, β = 82.83, γ = 83.49°. An X-ray diffraction data set was collected to 1.42 Å resolution with an Rmerge of 6.4%. Unexpectedly, a molecular-replacement solution was obtained using the crystal structure of Streptomyces lividans chloroperoxidase as a template, which shares 24% sequence identity to BtPhaZ. This is the first crystal structure of an intracellular poly(3-hydroxybutyrate) depolymerase.

  10. Screen of Bacillus thuringiensis toxins for transgenic rice to control Sesamia inferens and Chilo suppressalis.

    Science.gov (United States)

    Gao, Yulin; Hu, Yang; Fu, Qiang; Zhang, Jie; Oppert, Brenda; Lai, Fengxiang; Peng, Yufa; Zhang, Zhitao

    2010-09-01

    Transgenic rice to control stem borer damage is under development in China. To assess the potential of Bacillus thuringiensis (Bt) transgenes in stem borer control, the toxicity of five Bt protoxins (Cry1Aa, Cry1Ab, Cry1Ac, Cry1Ba and Cry1Ca) against two rice stem borers, Sesamia inferens (pink stem borer) and Chilo suppressalis (striped stem borer), was evaluated in the laboratory by feeding neonate larvae on artificial diets containing Bt protoxins. The results indicated that Cry1Ca exhibited the highest level of toxicity to both stem borers, with an LC(50) of 0.24 and 0.30 microg/g for C. suppressalis and S. inferens, respectively. However, S. inferens was 4-fold lower in susceptibility to Cry1Aa, and 6- and 47-fold less susceptible to Cry1Ab and Cry1Ba, respectively, compared to C. suppressalis. To evaluate interactions among Bt protoxins in stem borer larvae, toxicity assays were performed with mixtures of Cry1Aa/Cry1Ab, Cry1Aa/Cry1Ca, Cry1Ac/Cry1Ca, Cry1Ac/Cry1Ba, Cry1Ab/Cry1Ac, Cry1Ab/Cry1Ba, and Cry1Ab/Cry1Ca at 1:1 (w/w) ratios. All protoxin mixtures demonstrated significant synergistic toxicity activity against C. suppressalis, with values of 1.6- to 11-fold higher toxicity than the theoretical additive effect. Surprisingly, all but one of the Bt protoxin mixtures were antagonistic in toxicity to S. inferens. In mortality-time response experiments, S. inferens demonstrated increased tolerance to Cry1Ab and Cry1Ac compared to C. suppressalis when treated with low or high protoxin concentrations. The data indicate the utility of Cry1Ca protoxin and a Cry1Ac/Cry1Ca mixture to control both stem borer populations. Copyright 2010 Elsevier Inc. All rights reserved.

  11. High performance thermal insulation systems (HiPTI). Vacuum insulated products (VIP). Proceedings of the international conference and workshop

    Energy Technology Data Exchange (ETDEWEB)

    Zimmermann, M.; Bertschinger, H.

    2001-07-01

    These are the proceedings of the International Conference and Workshop held at EMPA Duebendorf, Switzerland, in January 2001. The papers presented at the conference's first day included contributions on the role of high-performance insulation in energy efficiency - providing an overview of available technologies and reviewing physical aspects of heat transfer and the development of thermal insulation as well as the state of the art of glazing technologies such as high-performance and vacuum glazing. Also, vacuum-insulated products (VIP) with fumed silica, applications of VIP systems in technical building systems, nanogels, VIP packaging materials and technologies, measurement of physical properties, VIP for advanced retrofit solutions for buildings and existing and future applications for advanced low energy building are discussed. Finally, research and development concerning VIP for buildings are reported on. The workshops held on the second day covered a preliminary study on high-performance thermal insulation materials with gastight porosity, flexible pipes with high performance thermal insulation, evaluation of modern insulation systems by simulation methods as well as the development of vacuum insulation panels with a stainless steel envelope.

  12. AAS 227: Day 3

    Science.gov (United States)

    Kohler, Susanna

    2016-01-01

    Editors Note:This week were at the 227th AAS Meeting in Kissimmee, FL. Along with several fellow authors from astrobites.com, I will bewritingupdates on selectedevents at themeeting and posting at the end of each day. Follow along here or atastrobites.com, or catch ourlive-tweeted updates from the@astrobites Twitter account. The usual posting schedule for AAS Nova will resumenext week.Welcome to Day 3 of the winter American Astronomical Society (AAS) meeting in Kissimmee! Several of us are attending the conference this year, and we will report highlights from each day here on astrobites. If youd like to see more timely updates during the day, we encourage you to follow @astrobites on twitter or search the #aas227 hashtag.Henry Norris Russell Lecture: Viewing the Universe with Infrared Eyes: The Spitzer Space Telescope (by Erika Nesvold)The Henry Norris Russell Award is the highest honor given by the AAS, for a lifetime of eminence in astronomy research. This years award went to Giovanni Fazio of the Harvard-Smithsonian Center for Astrophysics. Fazio became a leader in gamma ray astronomy before switching mid-career to the study of infrared astronomy, and he gave his award lecture on the latter subject, specifically on the Spitzer Space Telescope, one of the most successful infrared telescopes of all time.Artists rendering of the Spitzer space telescope. [NASA/JPL-Caltech]Spitzer has been operating for more than twelve years, and has resulted in over six thousand papers in refereed journals in that time. The telescope sits in an Earth-trailing orbit around the Sun, and is now farther from the Earth (1.4 AU) than the Earth is from the Sun. Fazio gave the audience a fascinating overview of the science done by Spitzer over more than a decade. One of the most productive areas of research for Spitzer is the study of exoplanets, which hadnt even been discovered when the Spitzer Telescope was first conceived. Spitzers high sensitivity and ability to observe exoplanets over

  13. NCBI nr-aa BLAST: CBRC-XTRO-01-1252 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available illus thuringiensis serovar israelensis ATCC 35646] gb|EAO53420.1| Collagen-like triple helix repeat protein... [Bacillus thuringiensis serovar israelensis ATCC 35646] ZP_00742312.1 1e-40 38% ...

  14. NCBI nr-aa BLAST: CBRC-GGAL-35-0149 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available illus thuringiensis serovar israelensis ATCC 35646] gb|EAO53420.1| Collagen-like triple helix repeat protein... [Bacillus thuringiensis serovar israelensis ATCC 35646] ZP_00742312.1 6e-30 33% ...

  15. Isolation and characterization of Bacillus thuringiensis strains active against Elasmopalpus lignosellus (Zeller, 1848 (Lepidoptera, Pyralidae

    Directory of Open Access Journals (Sweden)

    Janaina Zorzetti

    2017-08-01

    Full Text Available Elasmopalpus lignosellus (Zeller, 1848 (Lepidoptera, Pyralidae is an insect pest of 60 economically important crops, including sugarcane, wheat, soybean, rice, beans, sorghum, peanuts, and cotton. The aim of this work was to select and characterize Bacillus thuringiensis isolates with insecticidal activity against E. Lignosellus that could be used as an alternative method of control. Selective bioassays were done to evaluate the toxicity of 47 isolates against first instar larvae of E. lignosellus. For the most toxic bacterial strains, the lethal concentration (LC50 was estimated and morphological, biochemical and molecular methods were used to characterize the isolates. Among the 47 isolates tested, 12 caused mortality above 85% and showed LC50 values from 0.038E+8 to 0.855E+8 spores mL-1. Isolates BR83, BR145, BR09, BR78, S1534, and S1302 had the lowest LC50 values and did not differ from the standard HD-1 strain; the exception was BR83.The protein profiles produced bands with molecular masses of 60-130 kDa. The genes cry1, cry2, cry3, and cry11 were identified in the molecular characterization. The morphological analysis identified three different crystal inclusions: bipyramidal, spherical and cuboidal. Among the tested isolates, 12 isolates have potential for biotechnological control of E. Lignosellus by development of new biopesticides or genetically modified plants.

  16. Vital area identification software VIP for the physical protection of nuclear power plants

    International Nuclear Information System (INIS)

    Jung, Woo Sik; Park, Chang Kue; Yang, Joon Eon

    2004-01-01

    There are two major factors to be considered for the physical protection of nuclear power plants. They are a design basis threat (DBT) and the vital area identification (VAI). The DBT has been considered as 'the maximum credible threat.' The vital area is defined as 'an area inside a protected area containing equipment, systems or devices, or nuclear materials, the sabotage of which could directly or indirectly lead to unacceptable radiological consequences.' For the VAI of nuclear power plants, a software VIP (Vital area Identification Package based on PSA method) is being developed. The VIP is based on the current probabilistic safety assessment (PSA) techniques. The PSA method, including internal as well as external events, is known as the most complete and consistent method for identifying various accident sequences that might result in a core melt and radioactive material release to the environment. Thus, the VIP employs a fault tree analysis method in the PSA and utilizes the PSA results

  17. Spatial and temporal distribution of airborne Bacillus thuringiensis var. kurstaki during an aerial spray program for gypsy moth eradication.

    Science.gov (United States)

    Teschke, K; Chow, Y; Bartlett, K; Ross, A; van Netten, C

    2001-01-01

    We measured airborne exposures to the biological insecticide Bacillus thuringiensis var. kurstaki (Btk) during an aerial spray program to eradicate gypsy moths on the west coast of Canada. We aimed to determine whether staying indoors during spraying reduced exposures, to determine the rate of temporal decay of airborne concentrations, and to determine whether drift occurred outside the spray zone. During spraying, the average culturable airborne Btk concentration measured outdoors within the spray zone was 739 colony-forming units (CFU)/m3 of air. Outdoor air concentrations decreased over time, quickly in an initial phase with a half time of 3.3 hr, and then more slowly over the following 9 days, with an overall half-time of about 2.4 days. Inside residences during spraying, average concentrations were initially 2-5 times lower than outdoors, but at 5-6 hr after spraying began, indoor concentrations exceeded those outdoors, with an average of 244 CFU/m3 vs. 77 CFU/m3 outdoors, suggesting that the initial benefits of remaining indoors during spraying may not persist as outside air moves indoors with normal daily activities. There was drift of culturable Btk throughout a 125- to 1,000-meter band outside the spray zone where measurements were made, a consequence of the fine aerosol sizes that remained airborne (count median diameters of 4.3 to 7.2 microm). Btk concentrations outside the spray zone were related to wind speed and direction, but not to distance from the spray zone.

  18. Efficacy of some synthetic insecticides for control of cotton bollworms ...

    African Journals Online (AJOL)

    ... and Betsulfan at 3.2 l ha-1 recorded the highest and lowest yields, respectively. For effective control of cotton bollworms for maximum yield in the ecology, Thionex applied at 2.8 l ha-1 is recommended. Keywords: Control, cotton bollworms, efficacy, Ghana, synthetic insecticides. African Crop Science Journal, Vol. 20, No.

  19. Chemical composition of Rosmarinus officinalis and Lavandula stoechas essential oils and their insecticidal effects on Orgyia trigotephras (Lepidoptera: Lymantriidae

    Directory of Open Access Journals (Sweden)

    Ben Slimane Badreddine

    2015-01-01

    Full Text Available Objective: To evaluate toxic activities of essential oils obtained from Rosmarinus officinalis and Lavandula stoechas against the fourth larval instars of Orgyia trigotephras. Methods: A total of 1 200 larvae were divided into three groups-I, II, III. Group I was to investigate the effect of extracted essential oils from these aromatic plants as gastric disturbance. Bacillus thuringiensis was used as referencee and ethanol as control. Group II was used as contact action and Group III was used as fumigant action. For both Groups II and III, Decis was used as reference and ethanol as control. During the three experiments, the effect of essential oils on larvae was assessed. Results: The chemical composition of essential oils from two medicinal plants was determined, and their insecticidal effects on the fourth larval state of Orgyia trigotephras were assessed. They presented an insecticidal activity. Rosmarinus officinalis essential oil was less efficient compared to Lavandula stoechas. Conclusions: The relationship between the chemical composition and the biological activities is confirmed by the present findings. Therefore the potential uses of these essential oils as bioinsecticides can be considered as an alternative to the use of synthetic products.

  20. Transgenic plants expressing the AaIT/GNA fusion protein show increased resistance and toxicity to both chewing and sucking pests.

    Science.gov (United States)

    Liu, Shu-Min; Li, Jie; Zhu, Jin-Qi; Wang, Xiao-Wei; Wang, Cheng-Shu; Liu, Shu-Sheng; Chen, Xue-Xin; Li, Sheng

    2016-04-01

    The adoption of pest-resistant transgenic plants to reduce yield losses and decrease pesticide use has been successful. To achieve the goal of controlling both chewing and sucking pests in a given transgenic plant, we generated transgenic tobacco, Arabidopsis, and rice plants expressing the fusion protein, AaIT/GNA, in which an insecticidal scorpion venom neurotoxin (Androctonus australis toxin, AaIT) is fused to snowdrop lectin (Galanthus nivalis agglutinin, GNA). Compared with transgenic tobacco and Arabidopsis plants expressing AaIT or GNA, transgenic plants expressing AaIT/GNA exhibited increased resistance and toxicity to one chewing pest, the cotton bollworm, Helicoverpa armigera. Transgenic tobacco and rice plants expressing AaIT/GNA showed increased resistance and toxicity to two sucking pests, the whitefly, Bemisia tabaci, and the rice brown planthopper, Nilaparvata lugens, respectively. Moreover, in the field, transgenic rice plants expressing AaIT/GNA exhibited a significant improvement in grain yield when infested with N. lugens. This study shows that expressing the AaIT/GNA fusion protein in transgenic plants can be a useful approach for controlling pests, particularly sucking pests which are not susceptible to the toxin in Bt crops. © 2015 Institute of Zoology, Chinese Academy of Sciences.

  1. LARGE AREA MONITORING FOR PESTICIDAL TRANSGENIC CROPS: HOW SPECTRAL IMAGING MAY PLAY A ROLE

    Science.gov (United States)

    Crops genetically engineered to contain a bacterial gene that expresses an insecticidal protein from Bacillus thuringiensis are regulated by EPA under the Federal Insecticide Fungicide and Rodenticide Act (FIFRA). EPA has declared crops containing transgenic pesticidal traits to...

  2. NCBI nr-aa BLAST: CBRC-MMUS-02-0333 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MMUS-02-0333 ref|NP_054012.1| collagen-like protein [Bacillus thuringiensis serovar israel...ensis] gb|AAB16964.1| collagen-like protein [Bacillus thuringiensis serovar israelensis] NP_054012.1 7e-52 59% ...

  3. NCBI nr-aa BLAST: CBRC-GGAL-35-0414 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-GGAL-35-0414 ref|NP_054012.1| collagen-like protein [Bacillus thuringiensis serovar israel...ensis] gb|AAB16964.1| collagen-like protein [Bacillus thuringiensis serovar israelensis] NP_054012.1 2e-21 31% ...

  4. NCBI nr-aa BLAST: CBRC-OCUN-01-0845 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-OCUN-01-0845 ref|NP_054012.1| collagen-like protein [Bacillus thuringiensis serovar israel...ensis] gb|AAB16964.1| collagen-like protein [Bacillus thuringiensis serovar israelensis] NP_054012.1 4e-04 42% ...

  5. NCBI nr-aa BLAST: CBRC-PVAM-01-0280 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-PVAM-01-0280 ref|NP_054012.1| collagen-like protein [Bacillus thuringiensis serovar israel...ensis] gb|AAB16964.1| collagen-like protein [Bacillus thuringiensis serovar israelensis] NP_054012.1 5e-30 35% ...

  6. NCBI nr-aa BLAST: CBRC-TGUT-30-0011 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TGUT-30-0011 ref|NP_054012.1| collagen-like protein [Bacillus thuringiensis serovar israel...ensis] gb|AAB16964.1| collagen-like protein [Bacillus thuringiensis serovar israelensis] NP_054012.1 1e-49 43% ...

  7. NCBI nr-aa BLAST: CBRC-HSAP-02-0005 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-HSAP-02-0005 ref|NP_054012.1| collagen-like protein [Bacillus thuringiensis serovar israel...ensis] gb|AAB16964.1| collagen-like protein [Bacillus thuringiensis serovar israelensis] NP_054012.1 8e-05 30% ...

  8. VIP visit to CERN P5 CMS of Pakistan Science Members

    CERN Multimedia

    Hoch, Michael

    2012-01-01

    VIP visit to CERN P5 CMS of PAEC & JCPC Science Members List of PAEC Visitors: Dr. Badar Suleman - Member Science PAEC & Member of JCPC Dr. Waqar M. Butt - Member Engineering (Head of HMC3) Dr. Maqsood Ahmad - Chief Scientist (Head of Accelerator Project) List of CMS participants: Prof. Joseph Incandela, CMS Spokesperson Dr. Austin Ball, CMS Technical Coordinator Mr Andrzej Charkiewicz, CMS Resources Manager Dr. Michael Hoch, CMS Outreach activities, CMS photographer and guide Dr. Achille Petrilli, CMS Team Leader

  9. Identification and Characterization of Hyphantria cunea Aminopeptidase N as a Binding Protein of Bacillus thuringiensis Cry1Ab35 Toxin

    Directory of Open Access Journals (Sweden)

    Yakun Zhang

    2017-11-01

    Full Text Available The fall webworm, Hyphantria cunea (Drury is a major invasive pest in China. Aminopeptidase N (APN isoforms in lepidopteran larvae midguts are known for their involvement in the mode of action of insecticidal crystal (Cry proteins from Bacillus thuringiensis. In the present work, we identified a putative Cry1Ab toxin-binding protein, an APN isoform designated HcAPN3, in the midgut of H. cunea by ligand blot and mass spectrometry. HcAPN3 was highly expressed throughout all larval developmental stages and was abundant in the midgut and hindgut tissues. HcAPN3 was down-regulated at 6 h, then was up-regulated significantly at 12 h and 24 h after Cry1Ab toxin treatment. We expressed HcAPN3 in insect cells and detected its interaction with Cry1Ab toxin by ligand blot assays. Furthermore, RNA interference (RNAi against HcAPN3 using oral delivery and injection of double-stranded RNA (dsRNA resulted in a 61–66% decrease in transcript level. Down-regulating of the expression of HcAPN3 was closely associated with reduced susceptibility of H. cunea to Cry1Ab. In addition, the HcAPN3E fragment peptide expressed in Escherichia coli enhanced Cry1Ab toxicity against H. cunea larvae. This work represents the first evidence to suggest that an APN in H. cunea is a putative binding protein involved in Cry1Ab susceptibility.

  10. NCBI nr-aa BLAST: CBRC-OLAT-26-0107 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available fA [Bacillus thuringiensis serovar israelensis ATCC 35646] gb|EAO52941.1| SpoVID-dependent spore coat assemb...ly factor SafA [Bacillus thuringiensis serovar israelensis ATCC 35646] ZP_00742788.1 7e-18 33% ...

  11. Pharmacological characterization of VIP and PACAP receptors in the human meningeal and coronary artery

    DEFF Research Database (Denmark)

    Chan, Kayi Y; Baun, Michael; de Vries, René

    2011-01-01

    We pharmacologically characterized pituitary adenylate cyclase-activating polypeptides (PACAPs), vasoactive intestinal peptide (VIP) and the VPAC(1), VPAC(2) and PAC(1) receptors in human meningeal (for their role in migraine) and coronary (for potential side effects) arteries.......We pharmacologically characterized pituitary adenylate cyclase-activating polypeptides (PACAPs), vasoactive intestinal peptide (VIP) and the VPAC(1), VPAC(2) and PAC(1) receptors in human meningeal (for their role in migraine) and coronary (for potential side effects) arteries....

  12. VIP and PACAP display different vasodilatory effects in rabbit coronary and cerebral arteries

    DEFF Research Database (Denmark)

    Dalsgaard, Tórur; Hannibal, Jens; Fahrenkrug, Jan

    2003-01-01

    on the vasodilatory effects of these two peptides in cerebral and coronary vessels from female New Zealand White (NZW) rabbits.The localization and concentration of VIP and PACAP in cardiovascular tissue was evaluated using immunohistochemistry and radioimmunoassays. The vasodilatory effects of VIP and PACAP were...... investigated using myographs, allowing isometric tension recordings. In order to evaluate the influence of steroid hormones, the rabbits were ovariectomized and randomized to treatment for 4 weeks with 17beta-estradiol (E(2)), Norethindrone Acetate (NETA), E(2)+NETA or placebo. Ring segments of the posterior...

  13. The association between Interleukin (IL)-4 gene intron 3 VNTR polymorphism and alopecia areata (AA) in Turkish population.

    Science.gov (United States)

    Kalkan, Göknur; Karakus, Nevin; Baş, Yalçın; Takçı, Zennure; Ozuğuz, Pınar; Ateş, Omer; Yigit, Serbulent

    2013-09-25

    Alopecia areata (AA) is hypothesized to be an organ-specific autoimmune disease of hair follicles mediated by T cells. As immunological and genetic factors have been implicated in the pathogenesis of AA, the purpose of the present study was to investigate possible associations between the functional Interleukin (IL)-4 gene intron 3 VNTR polymorphism and AA susceptibility and disease progression in Turkish population. The study group consisted of 116 unrelated patients with AA and 125 unrelated healthy controls. Genomic DNA was isolated and IL-4 gene 70 bp VNTR polymorphism determined by using polymerase chain reaction (PCR) with specific primers. No association was observed between AA patients and controls according to genotype distribution (p=0.051). The allele distribution of IL-4 gene intron 3 VNTR polymorphism was statistically different between AA patients and control group (p=0.026). The frequency of P1 allele in patients was significantly higher than that in the control group. When the P2P2 genotype was compared with P1P2+P1P1 genotypes, a statistically significant difference was observed between patients and controls (p=0.036). Intron 3 VNTR polymorphism in the IL-4 gene was found to be associated with AA susceptibility in Turkish population. The results suggest that IL-4 VNTR polymorphism in the intron 3 region may be a risk factor for the development of AA among Turkish population. This is the first to report that intron 3 VNTR polymorphism in the IL-4 gene is associated with AA susceptibility. © 2013.

  14. [Potential of Bacillus thuringiensis israelensis Berliner for controlling Aedes aegypti].

    Science.gov (United States)

    Polanczyk, Ricardo Antonio; Garcia, Marcelo de Oliveira; Alves, Sérgio Batista

    2003-12-01

    The importance of the entomopathogenic bacterium Bacillus thuringiensis israelensis in the control of Aedes aegypti is presented. The use and potential of B. thuringiensis israelensis against the mosquito vector of dengue fever is described. Other aspects such as insect's resistance development against chemicals and advantages and constraints of using microbial control are discussed. Emphasis is given to the importance of the use of this bacterium in Brazil, which could contribute significantly to solving the mosquito problem without affecting the environment, humans and others invertebrate organisms in critical regions.

  15. Constitutive Activation of the Midgut Response to Bacillus thuringiensis in Bt-Resistant Spodoptera exigua

    NARCIS (Netherlands)

    Hernandez-Martinez, P.; Navarro-Cerrillo, G.; Caccia, S.; Maagd, de R.A.; Moar, W.J.; Ferre, J.; Escriche, B.; Herrero, S.

    2010-01-01

    Bacillus thuringiensis is the most effective microbial control agent for controlling numerous species from different insect orders. The main threat for the long term use of B. thuringiensis in pest control is the ability of insects to develop resistance. Thus, the identification of insect genes

  16. Efficacy of PermaNet® 2.0 and PermaNet® 3.0 against insecticide-resistant Anopheles gambiae in experimental huts in Côte d'Ivoire

    Directory of Open Access Journals (Sweden)

    Koffi Alphonsine A

    2011-06-01

    Full Text Available Abstract Background Pyrethroid resistance in vectors could limit the efficacy of long-lasting insecticidal nets (LLINs because all LLINs are currently treated with pyrethroids. The goal of this study was to evaluate the efficacy and wash resistance of PermaNet® 3.0 compared to PermaNet® 2.0 in an area of high pyrethroid in Côte d'Ivoire. PermaNet® 3.0 is impregnated with deltamethrin at 85 mg/m2 on the sides of the net and with deltamethrin and piperonyl butoxide on the roof. PermaNet® 2.0 is impregnated with deltamethrin at 55 mg/m2 across the entire net. Methods The study was conducted in the station of Yaokoffikro, in central Côte d'Ivoire. The efficacy of intact unwashed and washed LLINs was compared over a 12-week period with a conventionally-treated net (CTN washed to just before exhaustion. WHO cone bioassays were performed on sub-sections of the nets, using wild-resistant An. gambiae and Kisumu strains. Mosquitoes were collected five days per week and were identified to genus and species level and classified as dead or alive, then unfed or blood-fed. Results Mortality rates of over 80% from cone bioassays with wild-caught pyrethroid-resistant An. gambiae s.s were recorded only with unwashed PermaNet® 3.0. Over 12 weeks, a total of 7,291 mosquitoes were collected. There were significantly more An. gambiae s.s. and Culex spp. caught in control huts than with other treatments (P An. gambiae s.s and Culex spp, were lower for the control than for other treatments (P 0.05 except for unwashed PermaNet® 3.0 (P Conclusions This study showed that unwashed PermaNet® 3.0 caused significantly higher mortality against pyrethroid resistant An. gambiae s.s and Culex spp than PermaNet® 2.0 and the CTN. The increased efficacy with unwashed PermaNet® 3.0 over PermaNet® 2.0 and the CTN was also demonstrated by higher KD and mortality rates (KD > 95% and mortality rate > 80% in cone bioassays performed with wild pyrethroid-resistant An. gambiae s

  17. Effect of Stacked Insecticidal Cry Proteins from Maize Pollen on Nurse Bees (Apis mellifera carnica) and Their Gut Bacteria

    Science.gov (United States)

    Härtel, Stephan; Näther, Astrid; Dohrmann, Anja B.; Steffan-Dewenter, Ingolf; Tebbe, Christoph C.

    2013-01-01

    Honey bee pollination is a key ecosystem service to nature and agriculture. However, biosafety research on genetically modified crops rarely considers effects on nurse bees from intact colonies, even though they receive and primarily process the largest amount of pollen. The objective of this study was to analyze the response of nurse bees and their gut bacteria to pollen from Bt maize expressing three different insecticidal Cry proteins (Cry1A.105, Cry2Ab2, and Cry3Bb1). Naturally Cry proteins are produced by bacteria (Bacillus thuringiensis). Colonies of Apis mellifera carnica were kept during anthesis in flight cages on field plots with the Bt maize, two different conventionally bred maize varieties, and without cages, 1-km outside of the experimental maize field to allow ad libitum foraging to mixed pollen sources. During their 10-days life span, the consumption of Bt maize pollen had no effect on their survival rate, body weight and rates of pollen digestion compared to the conventional maize varieties. As indicated by ELISA-quantification of Cry1A.105 and Cry3Bb1, more than 98% of the recombinant proteins were degraded. Bacterial population sizes in the gut were not affected by the genetic modification. Bt-maize, conventional varieties and mixed pollen sources selected for significantly different bacterial communities which were, however, composed of the same dominant members, including Proteobacteria in the midgut and Lactobacillus sp. and Bifidobacterium sp. in the hindgut. Surprisingly, Cry proteins from natural sources, most likely B. thuringiensis, were detected in bees with no exposure to Bt maize. The natural occurrence of Cry proteins and the lack of detectable effects on nurse bees and their gut bacteria give no indication for harmful effects of this Bt maize on nurse honey bees. PMID:23533634

  18. Effect of stacked insecticidal Cry proteins from maize pollen on nurse bees (Apis mellifera carnica and their gut bacteria.

    Directory of Open Access Journals (Sweden)

    Harmen P Hendriksma

    Full Text Available Honey bee pollination is a key ecosystem service to nature and agriculture. However, biosafety research on genetically modified crops rarely considers effects on nurse bees from intact colonies, even though they receive and primarily process the largest amount of pollen. The objective of this study was to analyze the response of nurse bees and their gut bacteria to pollen from Bt maize expressing three different insecticidal Cry proteins (Cry1A.105, Cry2Ab2, and Cry3Bb1. Naturally Cry proteins are produced by bacteria (Bacillus thuringiensis. Colonies of Apis mellifera carnica were kept during anthesis in flight cages on field plots with the Bt maize, two different conventionally bred maize varieties, and without cages, 1-km outside of the experimental maize field to allow ad libitum foraging to mixed pollen sources. During their 10-days life span, the consumption of Bt maize pollen had no effect on their survival rate, body weight and rates of pollen digestion compared to the conventional maize varieties. As indicated by ELISA-quantification of Cry1A.105 and Cry3Bb1, more than 98% of the recombinant proteins were degraded. Bacterial population sizes in the gut were not affected by the genetic modification. Bt-maize, conventional varieties and mixed pollen sources selected for significantly different bacterial communities which were, however, composed of the same dominant members, including Proteobacteria in the midgut and Lactobacillus sp. and Bifidobacterium sp. in the hindgut. Surprisingly, Cry proteins from natural sources, most likely B. thuringiensis, were detected in bees with no exposure to Bt maize. The natural occurrence of Cry proteins and the lack of detectable effects on nurse bees and their gut bacteria give no indication for harmful effects of this Bt maize on nurse honey bees.

  19. Effect of stacked insecticidal Cry proteins from maize pollen on nurse bees (Apis mellifera carnica) and their gut bacteria.

    Science.gov (United States)

    Hendriksma, Harmen P; Küting, Meike; Härtel, Stephan; Näther, Astrid; Dohrmann, Anja B; Steffan-Dewenter, Ingolf; Tebbe, Christoph C

    2013-01-01

    Honey bee pollination is a key ecosystem service to nature and agriculture. However, biosafety research on genetically modified crops rarely considers effects on nurse bees from intact colonies, even though they receive and primarily process the largest amount of pollen. The objective of this study was to analyze the response of nurse bees and their gut bacteria to pollen from Bt maize expressing three different insecticidal Cry proteins (Cry1A.105, Cry2Ab2, and Cry3Bb1). Naturally Cry proteins are produced by bacteria (Bacillus thuringiensis). Colonies of Apis mellifera carnica were kept during anthesis in flight cages on field plots with the Bt maize, two different conventionally bred maize varieties, and without cages, 1-km outside of the experimental maize field to allow ad libitum foraging to mixed pollen sources. During their 10-days life span, the consumption of Bt maize pollen had no effect on their survival rate, body weight and rates of pollen digestion compared to the conventional maize varieties. As indicated by ELISA-quantification of Cry1A.105 and Cry3Bb1, more than 98% of the recombinant proteins were degraded. Bacterial population sizes in the gut were not affected by the genetic modification. Bt-maize, conventional varieties and mixed pollen sources selected for significantly different bacterial communities which were, however, composed of the same dominant members, including Proteobacteria in the midgut and Lactobacillus sp. and Bifidobacterium sp. in the hindgut. Surprisingly, Cry proteins from natural sources, most likely B. thuringiensis, were detected in bees with no exposure to Bt maize. The natural occurrence of Cry proteins and the lack of detectable effects on nurse bees and their gut bacteria give no indication for harmful effects of this Bt maize on nurse honey bees.

  20. Efficacy of PermaNet® 3.0 and PermaNet® 2.0 nets against laboratory-reared and wild Anopheles gambiae sensu lato populations in northern Tanzania.

    Science.gov (United States)

    Kweka, Eliningaya J; Lyaruu, Lucile J; Mahande, Aneth M

    2017-01-18

    Mosquitoes have developed resistance against pyrethroids, the only class of insecticides approved for use on long-lasting insecticidal nets (LLINs). The present study sought to evaluate the efficacy of the pyrethroid synergist PermaNet® 3.0 LLIN versus the pyrethroid-only PermaNet® 2.0 LLIN, in an East African hut design in Lower Moshi, northern Tanzania. In this setting, resistance to pyrethroid insecticides has been identified in Anopheles gambiae mosquitoes. Standard World Health Organization bioefficacy evaluations were conducted in both laboratory and experimental huts. Experimental hut evaluations were conducted in an area where there was presence of a population of highly pyrethroid-resistant An. arabiensis mosquitoes. All nets used were subjected to cone bioassays and then to experimental hut trials. Mosquito mortality, blood-feeding inhibition and personal protection rate were compared between untreated nets, unwashed LLINs and LLINs that were washed 20 times. Both washed and unwashed PermaNet® 2.0 and PermaNet® 3.0 LLINs had knockdown and mortality rates of 100% against a susceptible strain of An. gambiae sensu stricto. The adjusted mortality rate of the wild mosquito population after use of the unwashed PermaNet® 3.0 and PermaNet® 2.0 nets was found to be higher than after use of the washed PermaNet® 2.0 and PermaNet® 3.0 nets. Given the increasing incidence of pyrethroid resistance in An. gambiae mosquitoes in Tanzania, we recommend that consideration is given to its distribution in areas with pyrethroid-resistant malaria vectors within the framework of a national insecticide-resistance management plan.

  1. Willingness to pay for insecticide-treated nets in Berehet District, Amhara Region, Northern Ethiopia: implication of social marketing.

    Science.gov (United States)

    Aleme, Adisu; Girma, Eshetu; Fentahun, Netsanet

    2014-01-01

    Understanding the feasibility of achieving widespread coverage with Insecticide-Treated Nets has to be preceded by learning how people value the Insecticide-Treated Nets and estimating the potential demand and willingness to pay so that sustainability of the intervention can be assured. The objective of this study was to determine willingness to pay for Insecticide-Treated Nets among households in Berehet District, Northern Ethiopia. A community-based cross-sectional study was conducted using both quantitative and qualitative methods in five randomly selected Kebeles from January-February 2012. Open ended contingent valuation technique with follow-up method was used. Qualitative data were collected through focus group discussions and observation methods. Binary logistic regression was used to determine the association between dependent and independent variables. The average number of individuals per Insecticide-Treated Nets was 3.83. Nearly 68.5% persons had willingness to buy Insecticide-Treated Nets if they have access to these Nets. The median maximum price a person is willingness to pay for blue rectangular Insecticide-Treated Net was 20 ETB. People had willingness to pay 30 ETB for blue and white conical insecticide-treated nets. Working on knowledge of malaria (OR=0.68, CI (0.47, 0.98; ppay Insecticide-Treated Nets. Respondents who prefer Kebele/place/ to buy Insecticide-Treated Net for rectangular shape had a significant association with a willingness to pay for Insecticide-Treated Nets (OR=1.92, CI= 1.07-3.92). Promotions, products, price and place had significant association with willingness to pay for Insecticide-Treated Nets. Designing a social marketing strategy helps ensure sustainable supply of Insecticide-Treated Nets and proper use of Insecticide-Treated Nets.

  2. NCBI nr-aa BLAST: CBRC-CBRE-01-0086 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-CBRE-01-0086 ref|ZP_00739524.1| Integral membrane protein [Bacillus thuringiensis serovar israel...ensis ATCC 35646] gb|EAO56211.1| Integral membrane protein [Bacillus thuringiensis serovar israelensis ATCC 35646] ZP_00739524.1 0.038 25% ...

  3. NCBI nr-aa BLAST: CBRC-PMAR-01-0648 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-PMAR-01-0648 ref|ZP_00741686.1| Sodium/pantothenate symporter [Bacillus thuringiensis serovar israel...ensis ATCC 35646] gb|EAO54033.1| Sodium/pantothenate symporter [Bacillus thuringiensis serovar israelensis ATCC 35646] ZP_00741686.1 0.14 31% ...

  4. Effect of inherited sterility and bacillus thuringiensis on mortality and reproduction of phthorimaea opercullela zeller (lepidoptera: gelechidae)

    International Nuclear Information System (INIS)

    Makee, H.; Tlas, M. D.; Amer, S.; Abdulla, J.

    2008-01-01

    The effect of a commercial formulation of Bacillus thuringiensis (Dipel 2X) upon F 1 progeny of irradiated and unirradiated phthorimaea operculella male parents was investigated. F 1 progeny of irradiated parents was more susceptible to B. thuringiensis than that of unirradiated parents. A combination of irradiation and B. thuringiensis led to higher mortality in F 1 progeny of P. operculella. The LC 50 was 0.406 g/100ml for F 1 progeny of unirradiated parents, but 0.199 g/100ml for those of irradiated parents. There was a great reduction in the pupal weight, fecundity and egg hatchability of F 1 progeny of irradiated patents compared to those unirradiated parents. Such reduction was increased by applying higher concentration of B. thuringiensis. A combination between inherited sterility technique and B. thuringiensis application could give a good controlling result against P. operculella. (author)

  5. Germination and conjugation of Bacillus thuringiensis subsp. israelensis in the intestine of gnotobiotic rats

    DEFF Research Database (Denmark)

    Wilcks, Andrea; Ørum-Smidt, Lasse; Bahl, Martin Iain

    2008-01-01

    of growth and sporulation of vegetative cells. For the first time conjugative plasmid transfer in a mammalian intestinal tract was shown between two B. thuringiensis strains. Significance and Impact of the Study: Strains of B. thuringiensis are used worldwide to combat insect pests, and this study brings...

  6. Mechanism and DNA-based detection of field-evolved resistance to transgenic Bt corn in fall armyworm (Spodoptera frugiperda)

    Science.gov (United States)

    Evolution of resistance threatens sustainability of transgenic crops producing insecticidal proteins from the bacterium Bacillus thuringiensis (Bt). The fall armyworm is a devastating pest controlled by transgenic Bt corn producing the Cry1Fa insecticidal protein. However, fall armyworm populations ...

  7. Degradation of Insecticides in Poultry Manure: Determining the Insecticidal Treatment Interval for Managing House Fly (Diptera: Muscidae) Populations in Poultry Farms.

    Science.gov (United States)

    Ong, Song-Quan; Ab Majid, Abdul Hafiz; Ahmad, Hamdan

    2016-04-01

    It is crucial to understand the degradation pattern of insecticides when designing a sustainable control program for the house fly, Musca domestica (L.), on poultry farms. The aim of this study was to determine the half-life and degradation rates of cyromazine, chlorpyrifos, and cypermethrin by spiking these insecticides into poultry manure, and then quantitatively analyzing the insecticide residue using ultra-performance liquid chromatography. The insecticides were later tested in the field in order to study the appropriate insecticidal treatment intervals. Bio-assays on manure samples were later tested at 3, 7, 10, and 15 d for bio-efficacy on susceptible house fly larvae. Degradation analysis demonstrated that cyromazine has the shortest half-life (3.01 d) compared with chlorpyrifos (4.36 d) and cypermethrin (3.75 d). Cyromazine also had a significantly greater degradation rate compared with chlorpyrifos and cypermethrin. For the field insecticidal treatment interval study, 10 d was the interval that had been determined for cyromazine due to its significantly lower residue; for ChCy (a mixture of chlorpyrifos and cypermethrin), the suggested interval was 7 d. Future work should focus on the effects of insecticide metabolites on targeted pests and the poultry manure environment.

  8. Ecdysone Agonist: New Insecticides with Novel Mode of Action

    Directory of Open Access Journals (Sweden)

    Y. Andi Trisyono

    2002-12-01

    Full Text Available Development of insect resistance to insecticide has been the major driving force for the development of new insecticides. Awareness and demand from public for more environmentally friendly insecticides have contributed in shifting the trend from using broad spectrum to selective insecticides. As a result, scientists have looked for new target sites beyond the nervous system. Insect growth regulators (IGRs are more selective insecticides than conventional insecticides, and ecdysone agonists are the newest IGRs being commercialized, e.g. tebufenozide, methoxyfenozide, and halofenozide. Ecdysone agonists bind to the ecdysteroid receptors, and they act similarly to the molting hormone 20-hydroxyecdysone. The binding provides larvae or nymphs with a signal to enter a premature and lethal molting cycle. In addition, the ecdysone agonists cause a reduction in the number of eggs laid by female insects. The ecdysone agonists are being developed as selective biorational insecticides. Tebufenozide and methoxyfenozide are used to control lepidopteran insect pests, whereas halofenozide is being used to control coleopteran insect pests. Their selectivity is due to differences in the binding affinity between these compounds to the receptors in insects from different orders. The selectivity of these compounds makes them candidates to be used in combinations with other control strategies to develop integrated pest management programs in agricultural ecosystems. Key words: new insecticides, selectivity, ecdysone agonists

  9. Corpo e masculinidade na revista VIP Exame Body and masculinity in the magazine VIP Exame

    Directory of Open Access Journals (Sweden)

    Marko Monteiro

    2001-01-01

    Full Text Available Nesse artigo elaboro comentários teóricos acerca de novas formas de se vivenciar a corporalidade presentes contemporaneamente, a partir da análise da revista masculina VIP Exame. O texto baseia-se numa pesquisa que incluiu uma observação dentro da redação da revista e análise de materiais impressos, centrando-se na forma como a revista aborda o corpo masculino e coloca a preocupação com a aparência como importante para uma masculinidade bem sucedida. Com base nestes dados busco avaliar, a partir de perspectivas feministas e sobre o corpo, a influência do valor dado ao corpo e ao seu monitoramento reflexivo na constituição de identidades de gênero.In this article I elaborate theoretical comments on the subject of corporeality and the new forms of experiencing the body that are present in contemporary times. On the basis of a research with a Brazilian men's magazine, VIP Exame, which involved the analysis of issues of the magazine as well as an observation of the working process of the reporters and journalists, I focus my comments on the way the masculine body is treated by the magazine and how a discourse emerges on masculinity where attention to "good looks" becomes an important feature of a successful masculinity. On the basis of these observations I go on to discuss feminist and other theories of the subject, in order to understand how the reflexive monitoring of the body is increasingly important in the constitution of gender identities.

  10. Proteolytic processing of Bacillus thuringiensis Vip3A proteins by two Spodoptera species

    Czech Academy of Sciences Publication Activity Database

    Caccia, S.; Chakroun, M.; Vinokurov, Konstantin; Ferré, J.

    2014-01-01

    Roč. 67, AUG 2014 (2014), s. 76-84 ISSN 0022-1910 R&D Projects: GA ČR(CZ) GAP502/11/1471 Grant - others:Spanish Ministry of Science and Innovation(ES) AGL2009-13340-C02-01; Generalitat Valenciana and by European FEDER funds (ES) ACOMP/2009/313; Generalitat Valenciana and by European FEDER funds (ES) PROMETEO 2011/044; Marie Curie Grant(ES) PIEF-GA-2008-219993; European Union FP7(US) MOBITAG, GA 229518 Institutional support: RVO:60077344 Keywords : mode of action * serine peptidases * midgut peptidases Subject RIV: ED - Physiology Impact factor: 2.470, year: 2014 http://www.sciencedirect.com/science/article/pii/S0022191014001231

  11. NCBI nr-aa BLAST: CBRC-PMAR-01-0052 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-PMAR-01-0052 ref|ZP_00741639.1| Spore germination protein LA [Bacillus thuringiensis serovar israel...ensis ATCC 35646] gb|EAO54096.1| Spore germination protein LA [Bacillus thuringiensis serovar israelensis ATCC 35646] ZP_00741639.1 0.34 24% ...

  12. NCBI nr-aa BLAST: CBRC-OANA-01-2117 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-OANA-01-2117 ref|ZP_00742975.1| Hypothetical protein RBTH_01502 [Bacillus thuringiensis serovar israel...ensis ATCC 35646] gb|EAO52751.1| Hypothetical protein RBTH_01502 [Bacillus thuringiensis serovar israelensis ATCC 35646] ZP_00742975.1 1e-06 40% ...

  13. NCBI nr-aa BLAST: CBRC-MDOM-02-0482 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MDOM-02-0482 ref|ZP_04075807.1| Prophage LambdaBa01, membrane protein [Bacillu...s thuringiensis IBL 200] gb|EEM92524.1| Prophage LambdaBa01, membrane protein [Bacillus thuringiensis IBL 200] ZP_04075807.1 2e-11 18% ...

  14. NCBI nr-aa BLAST: CBRC-TTRU-01-0303 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TTRU-01-0303 ref|ZP_04119696.1| O-acetyl transferase [Bacillus thuringiensis serovar pakistan...i str. T13001] gb|EEM48546.1| O-acetyl transferase [Bacillus thuringiensis serovar pakistani str. T13001] ZP_04119696.1 0.008 23% ...

  15. NCBI nr-aa BLAST: CBRC-TTRU-01-0284 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TTRU-01-0284 ref|ZP_04119696.1| O-acetyl transferase [Bacillus thuringiensis serovar pakistan...i str. T13001] gb|EEM48546.1| O-acetyl transferase [Bacillus thuringiensis serovar pakistani str. T13001] ZP_04119696.1 0.009 26% ...

  16. NCBI nr-aa BLAST: CBRC-VPAC-01-0194 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-VPAC-01-0194 ref|ZP_04121235.1| Transporter, MFS superfamily [Bacillus thuringiensis serovar pakistan...i str. T13001] gb|EEM47055.1| Transporter, MFS superfamily [Bacillus thuringiensis serovar pakistani str. T13001] ZP_04121235.1 0.99 23% ...

  17. 40 CFR 180.1107 - Delta endotoxin of Bacillus thuringiensis variety kurstaki encapsulated into killed Pseudomonas...

    Science.gov (United States)

    2010-07-01

    ... thuringiensis variety kurstaki encapsulated into killed Pseudomonas fluorescens; exemption from the requirement... killed Pseudomonas fluorescens; exemption from the requirement of a tolerance. The delta endotoxin of Bacillus thuringiensis variety kurstaki encapsulated into killed Pseudomonas fluorescens is exempt from the...

  18. Insecticide susceptibility of Aedes aegypti and Aedes albopictus in Central Africa

    Directory of Open Access Journals (Sweden)

    Nwane Philippe

    2011-05-01

    Full Text Available Abstract Background Aedes aegypti (Linnaeus, 1762 and Aedes albopictus (Skuse, 1894 are the main vectors of dengue (DENV and chikungunya (CHIKV viruses worldwide. As there is still no vaccine or specific treatment for DENV and CHIKV, vector control remains the cornerstone of prevention and outbreak control. Unfortunately, vector control programs are facing operational challenges with mosquitoes becoming resistant to commonly used insecticides in several areas through the world. Throughout Central Africa no recent data are available susceptible/resistant status of either vector species since the introduction/arrival of Ae. albopictus in this area. We therefore studied the level of resistance of these two major vectors to insecticides commonly used in Africa for mosquito control. Results Aedes aegypti and Ae. albopictus were sampled in six urban localities of Cameroon (Garoua, Bertoua, Yaoundé, Bafia, Buea and Gabon (Libreville. Larval bioassays, carried out to determine the lethal concentrations (LC50 and LC95 and resistance ratios (RR50 and RR95 suggested that both vector species were susceptible to Bti (Bacillus thuringiensis var israeliensis and temephos. Bioassays were also performed on adults using WHO diagnostic test kits to assess phenotypic resistance to deltamethrin, DDT, fenitrothion and propoxur. These experiments showed that one population of Ae. aegypti (Libreville and two populations of Ae. albopictus (Buea and Yaoundé were resistant to DDT (mortality 36% to 71%. Resistance to deltamethrin was also suspected in Ae. albopictus from Yaoundé (83% mortality. All other field mosquito populations were susceptible to deltamethrin, DDT, fenitrothion and propoxur. No increase in the knockdown times (Kdt50 and Kdt95 was noted in the Yaoundé resistant population compared to other Ae. albopictus populations, suggesting the possible involvement of metabolic resistance to deltamethrin and DDT. Conclusion In view of the recent increase in

  19. NCBI nr-aa BLAST: CBRC-TTRU-01-1232 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available thuringiensis serovar pakistani str. T13001] gb|EEM48940.1| hypothetical protein bthur0005_11430 [Bacillus thuringiensis serovar pakistani str. T13001] ZP_04119374.1 0.031 24% ... ...CBRC-TTRU-01-1232 ref|ZP_04119374.1| hypothetical protein bthur0005_11430 [Bacillus

  20. NCBI nr-aa BLAST: CBRC-TTRU-01-1317 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available thuringiensis serovar pakistani str. T13001] gb|EEM45303.1| hypothetical protein bthur0005_48630 [Bacillus thuringiensis serovar pakistani str. T13001] ZP_04123033.1 0.059 29% ... ...CBRC-TTRU-01-1317 ref|ZP_04123033.1| hypothetical protein bthur0005_48630 [Bacillus

  1. Genetic engineering of cotton with a novel cry2AX1 gene to impart insect resistance against Helicoverpa armigera

    Directory of Open Access Journals (Sweden)

    Karunamurthy Dhivya

    2016-09-01

    Full Text Available Embryogenic calli of cotton (Coker310 were cocultivated with the Agrobacterium tumefaciens strain LBA4404 harbouring the codon-optimised, chimeric cry2AX1 gene consisting of sequences from cry2Aa and cry2Ac genes isolated from Indian strains of Bacillus thuringiensis. Forty-eight putative transgenic plants were regenerated, and PCR analysis of these plants revealed the presence of the cry2AX1 gene in 40 plants. Southern blot hybridisation analysis of selected transgenic plants confirmed stable T-DNA integration in the genome of transformed plants. The level of Cry2AX1 protein expression in PCR positive plants ranged from 4.9 to 187.5 ng g-1 of fresh tissue. A transgenic cotton event, TP31, expressing the cry2AX1 gene showed insecticidal activity of 56.66 per cent mortality against Helicoverpa armigera in detached leaf disc bioassay. These results indicate that the chimeric cry2AX1 gene expressed in transgenic cotton has insecticidal activity against H. armigera.

  2. Assessment of Adiantum incisum, Alternanthera pungens and Trichodesma indicum as bio-insecticides against stored grain pests

    International Nuclear Information System (INIS)

    Safdar, N.; Yasmin, A.

    2017-01-01

    This study investigated the insecticidal potential of Adiantum incisum Forssk (Pteridaceae), Alternanthera pungens Kunth (Amaranthaceae) and Trichodesma indicum L. (Boraginaceae). Aqueous, methanolic and n-hexane extracts of whole plants (roots, stem and leaves) were prepared using maceration technique. Insecticidal activities of aqueous, methanolic and n-hexane extracts of three plants (10, 20 and 30 mg/mL) were evaluated by impregnated filter paper insecticidal assay. Methanol and hexane extracts of Adiantum incisum gave effective LD 50 (15.3 mg/mL) against Callosobruchus chinensis and Sitophilus oryzae (LD5022 mg/mL) after 24 hours respectively. Present research elucidates the important phytochemicals (alkaloids, saponins and tannins) in three plants by FTIR and good insecticidal activity (LD50 < 25 mg/mL in 24 hours) against Callosobruchus chinensis and Sitophilus oryzae. Current study promotes further investigations for using these plant extracts as anti-feedants, repellents, fumigants and formulation of non-toxic bio-insecticides. (author)

  3. Insecticide control of vector-borne diseases: when is insecticide resistance a problem?

    Directory of Open Access Journals (Sweden)

    Ana Rivero

    Full Text Available Many of the most dangerous human diseases are transmitted by insect vectors. After decades of repeated insecticide use, all of these vector species have demonstrated the capacity to evolve resistance to insecticides. Insecticide resistance is generally considered to undermine control of vector-transmitted diseases because it increases the number of vectors that survive the insecticide treatment. Disease control failure, however, need not follow from vector control failure. Here, we review evidence that insecticide resistance may have an impact on the quality of vectors and, specifically, on three key determinants of parasite transmission: vector longevity, competence, and behaviour. We argue that, in some instances, insecticide resistance is likely to result in a decrease in vector longevity, a decrease in infectiousness, or in a change in behaviour, all of which will reduce the vectorial capacity of the insect. If this effect is sufficiently large, the impact of insecticide resistance on disease management may not be as detrimental as previously thought. In other instances, however, insecticide resistance may have the opposite effect, increasing the insect's vectorial capacity, which may lead to a dramatic increase in the transmission of the disease and even to a higher prevalence than in the absence of insecticides. Either way-and there may be no simple generality-the consequence of the evolution of insecticide resistance for disease ecology deserves additional attention.

  4. Bacillus thuringiensis and its application in agriculture

    African Journals Online (AJOL)

    USER

    2010-04-05

    Apr 5, 2010 ... Key words: Bacillus thuringiensis, endotoxins, crop plants. INTRODUCTION ..... of resistance in the pest and unfavorable interactions with beneficial .... with slower resistance evolution in North Carolina compared to .... level of 0.18% cross pollination in the experimental rice lines. .... Ecology and Safety.

  5. Interação entre inimigos naturais: Trichogramma e Bacillus thuringiensis no controle biológico de pragas agrícolas = Interaction between natural enemies: Trichogramma and Bacillus thuringiensis in pest control

    Directory of Open Access Journals (Sweden)

    Ricardo Antonio Polanczyk

    2006-04-01

    Full Text Available O objetivo deste trabalho foi avaliar os efeitos adversos de Bacillus thuringiensis (Bt nos parasitóides de ovos Trichogramma pratissolii e Trichogramma pretiosum. Suspensões de 6 isolados de Bt: E-3, E-10, E-15, E-16, E-19, E-20 e Bt kurstaki foram misturadas com mel (1:1, como fonte de alimento, e cartelas com ovos de Anagasta kuehniella foram fornecidas para o parasitismo. O experimento foi mantido em BOD a 25 + 1ºC, umidade relativa de 70+ 10% e fotofase de 14 horas. Foram analisados a longevidade, os parasitismos diário, acumulado e total. Não houve influência dos tratamentos sobre a longevidade e todos ostratamentos em T. pratissolii e E-3, E-10, E-16 e E- 19 em T. pretiosum diminuíram o tempo necessário para os parasitóides atingirem 80% de parasitismo. Apesar dessa aceleração do parasitismo, o Bt não influenciou o total de ovos parasitados, mostrando-se como tática de controle que pode ser usada com Trichogramma em programas de MIP.The goal of this work was to detect the possible side-effectsprovoked by Bacillus thuringiensis (Bt strains on eggs parasitoids Thichogramma pratissolii and Trichogramma pretiosum. Six strains of Bt: E-3, E-10, E-15, E-16, E-19, and E-20 and Bt kurstaki were assayed in honey drop to stimulate the feeding of the parasitoids in each treatment and offered simultaneously displays with Anagasta kuehniella eggs. The experiment was maintained in a climatized chamber at 25+ 1ºC, RH 70+ 10%, and photophase of 14 hours. The longevity, the daily, the accumulated, and the total parasitism were observed. No treatments affected the longevity of both parasitoids. All treatments in T. pratissolii and E-3, E-10, E-16, and E-19 in T. pretiosum decreased the time necessary to reach 80% of parasitism. The lack of deleterious effects of Bt on these parasitoids suggests that these natural enemies can be used together in IPM programs.

  6. Mapping VIPS concepts for nursing interventions to the ISO reference terminology model for nursing actions: A collaborative Scandinavian analysis

    DEFF Research Database (Denmark)

    Ehnfors, Margareta; Angermo, Lilly Marit; Berring, Lene

    2006-01-01

    analyzed the VIPS model's concepts for nursing interventions using prototypical examples of nursing actions, involving 233 units of analyses, and collaborated in mapping the two models. All nursing interventions in the VIPS model comprise actions and targets, but a few lack explicit expressions of means......The aims of this study were to analyze the coherence between the concepts for nursing interventions in the Swedish VIPS model for nursing recording and the ISO Reference Terminology Model for Nursing Actions and to identify areas in the two models for further development. Seven Scandinavian experts....... In most cases, the recipient of care is implicit. Expressions for the aim of an action are absent from the ISO model. By this mapping we identified areas for future development of the VIPS model and the experience from nursing terminology work in Scandinavia can contribute to the international...

  7. Laboratory investigations of insecticide impregnated materials for the control of New World screwworm flies

    International Nuclear Information System (INIS)

    Harris, E.G.; Brown, M.; Smith, S.

    1992-01-01

    In laboratory tests, New World screwworm flies were found to be at least three orders of magnitude less susceptible to the insecticides deltamethrin, alphacypermethrin and cyfluthrin than are tsetse flies. Deltamethrin was the most toxic of the three insecticides to screwworm flies. For topical application, the LD50s for deltamethrin 20% suspension concentrate were 33 ng and 25 ng for male and female screwworm flies respectively, compared with 0.04 ng for tsetse, G.m. morsitans. In various tests simulating contact of screwworm flies with cloth or netting targets impregnated with insecticide, 100% kill was only achieved with 3.2% deltamethrin and contact times of at least 10 sec, although 100% knockdown for up to 24 hours was obtained with lower concentrations. No repellent effect was observed at the higher concentrations. Cloth targets impregnated with a high dose of insecticide and baited with an attractant could be effective against NWS flies, especially if after ''knockdown'' flies are removed by predators. 1 fig., 9 tabs

  8. Volta potential of clad AA2024 aluminium after exposure to CeCl3 solution

    International Nuclear Information System (INIS)

    Andreatta, F.; Druart, M.-E.; Marin, E.; Cossement, D.; Olivier, M.-G.; Fedrizzi, L.

    2014-01-01

    Highlights: • Alkaline etch of clad AA2024 enhances precipitation of Ce compounds. • Exposure to CeCl 3 solution decreases Volta potential of alkaline etched substrate. • Ce compounds reduce the driving force for initiation of localized attack. - Abstract: AA2024 clad with AA1050 was immersed in CeCl 3 solution to promote deposition of cerium species. The deposition occurs on the entire sample surface for the alkaline etched substrate, while it is very limited for the degreased substrate. The surface potential (Volta potential) was investigated by scanning Kelvin probe force microscopy after different immersion times in CeCl 3 solution. The preferential deposition of Ce compounds at Al–Fe intermetallic sites progressively reduces their Volta potential difference relative to the matrix in the alkaline etched substrate. This reduces the susceptibility to localized attack of the intermetallics as proven by potentiodynamic polarization measurements

  9. Preliminary investigations reveal that Bacillus thuringiensis δ ...

    African Journals Online (AJOL)

    The imminent introduction of transgenic crops into Kenya requires a rigorous assessment of the potential risks involved. This study focused on the possible effect of Bacillus thuringiensisδ-endotoxin [CryIA(c)] on arbuscular mycorrhizal fungi (AMF) associated with sorghum. In green house experiments, sorghum seedlings ...

  10. Susceptibilidade de larvas de Cerotoma arcuata Olivier (Coleoptera: Chrysomelidae a Beauveria bassiana (Bals. Vuillemin, Metarhizium anisopliae (Metsch. Sorokin e Bacillus thuringiensis Berliner Susceptibility of Cerotoma arcuata Olivier (Coleoptera: Chrysomelidae larvae to Beauveria bassiana (Bals. Vuillemin, Metarhizium anisopliae (Metsch. Sorokin and Bacillus thuringiensis Berliner

    Directory of Open Access Journals (Sweden)

    Maria Lucia França Teixeira

    2007-02-01

    Full Text Available Larvas de 2° instar de Cerotoma arcuata foram avaliadas em relação à susceptibilidade aos fungos entomopatogênicos Beauveria bassiana, Metarhizium anisopliae e a bactéria Bacillus thuringiensis com as toxinas Cry3. Os insetos adultos foram mantidos em gaiolas e alimentados com plântulas de feijão (Phaseolus vulgaris L. e as larvas em "gerbox" com cotilédones de plântulas de feijão recém-germinadas. Das oito estirpes de B. bassiana avaliadas, CG 156 e CG 213 causaram 100% de mortalidade das larvas, as duas estirpes de M. anisopliae CG 210 e CG 321 foram patogênicas, eliminando 80 e 100% das larvas de C. arcuata, e, das cinco estirpes de B. thuringiensis testadas, o isolado CG 940 causou 70% de mortalidade das larvas.Second instar larvae of Cerotoma arcuata were evaluated concerning the susceptibility to fungi Beauveria bassiana and Metarhizium anisopliae and Bacillus thuringiensis strains containing Cry3 toxin. Adults of C. arcuata were kept in large cages and fed on bean seedlings and the larvae were reared in ‘gearbox’ feeding on germinated Phaseolus bean cotyledons. Strains CG 156 and CG 213 of B. bassiana killed 100% of the insect larvae and strains CG 210 and CG 321 of M. anisopliae killed 80 and 100% of the insect larvae. Strain CG 940 of B. thuringiensis killed 70% of the insect larvae.

  11. A Voltage Instability Predictor Using Local Area Measurements. VIP++

    Energy Technology Data Exchange (ETDEWEB)

    Warland, Leif

    2002-07-01

    There has been a pressure to operate power systems closer to their security limits. This has partially been due to financial imperatives following the deregulating of markets. Other practical difficulties have been obtaining authorization from regulatory bodies to build power plants and transmission lines. In this situation it is essential to monitor the system and to have tools that can predict the distance to the point of collapse (PoC). Much effort has been put into research of the phenomenon voltage collapse, and many approaches have been explored. Both dynamic and steady-state behavior have been studied thoroughly, though very few protection and control schemes have been implemented. In this dissertation the possibility of an index based on local area measurements have been explored. Voltage stability can be classified as either a transient or a long-term stability problem, and the index proposed in this dissertation is based on long-term dynamics. The VIP algorithm is a method that uses the maximum load ability of a transmission network as the PoC, thus by estimating a Thevenin equivalent the method can track the distance to the PoC as this occurs when the two impedances are equal in absolute value. The problem of the VIP algorithm is that it is based on a system with two equations and four unknowns, thus it is not observable. In order to make it observable the assumption of constant Thevenin equivalent between two sets of measurements is made. When this is not the case the method will estimate a Thevenin impedance of the same size as the measured load impedance, but with a negative sign. Changes in the Thevenin equivalent can be traced to both angle variation in a remote generator area or to variations in load impedances on nearby buses. The problem of angle variations can be mitigated by the selection of an appropriate reference bus. A method to solve the second problem, of variation in load admittances, has been proposed in this dissertation and given the

  12. Effects of irrigation levels on interactions among Lygus hesperus (Hemiptera: Miridae), insecticides, and predators in cotton.

    Science.gov (United States)

    Asiimwe, Peter; Naranjo, Steven E; Ellsworth, Peter C

    2014-04-01

    Variation in plant quality and natural enemy abundance plays an important role in insect population dynamics. In manipulative field studies, we evaluated the impact of varying irrigation levels and insecticide type on densities of Lygus hesperus Knight and the arthropod predator community in cotton. Three watering levels were established via irrigations timed according to three levels of percent soil water depletion (SWD): 20, 40, or 60, where 40% SWD is considered standard grower practice, 60% represents a deficit condition likely to impose plant productivity losses, and 20% represents surplus conditions with likely consequences on excessive vegetative plant production. The two key L. hesperus insecticides used were the broad-spectrum insecticide acephate and the selective insecticide flonicamid, along with an untreated check. We hypothesized that densities of L. hesperus and its associated predators would be elevated at higher irrigation levels and that insecticides would differentially impact L. hesperus and predator dynamics depending on their selectivity. L. hesperus were more abundant at the higher irrigation level (20% SWD) but the predator densities were unaffected by irrigation levels. Both L. hesperus and its predators were affected by the selectivity of the insecticide with highest L. hesperus densities and lowest predator abundance where the broad spectrum insecticide (acephate) was used. There were no direct interactions between irrigation level and insecticides, indicating that insecticide effects on L. hesperus and its predators were not influenced by the irrigation levels used here. The implications of these findings on the overall ecology of insect-plant dynamics and yield in cotton are discussed.

  13. Methodology for fast evaluation of Bacillus thuringiensis crystal protein content

    Directory of Open Access Journals (Sweden)

    Alves Lúcia M. Carareto

    2000-01-01

    Full Text Available The development of the production and use of Bacillus thuringiensis in Brazil at a commercial scale faces certain difficulties, among them the establishment of efficient methodologies for the quantitation of toxic products to be commercialized. Presently, the amount of toxin is given in percentage by analyzing the samples total protein content. Such methodology however, does not measure the actual amount of active protein present in the product, since most strains express different endotoxin genes and might even produce b-toxin. Since the various types of toxins exhibit different antigenic characteristics, this work has as objective the utilization of fast immunological techniques to quantify the level of crystal protein. Crystal protein produced by a subspecies of Bacillus thuringiensis var. israelensis was purified by ultracentrifugation and utilized to immunize rabbits and to produce hiperimmune sera. Such sera were latter used to evaluate the level of proteins on commercial bioinsecticide and on laboratory cultures of B. thuringiensis through the immunodot technique. The results were obtained by comparison of data obtained from reactions with known concentrations of crystal protein permitting to evaluate the level of such protein on various materials.

  14. 40 CFR 180.1108 - Delta endotoxin of Bacillus thuringiensis variety San Diego encapsulated into killed Pseudomonas...

    Science.gov (United States)

    2010-07-01

    ... thuringiensis variety San Diego encapsulated into killed Pseudomonas fluorescens; exemption from the requirement... into killed Pseudomonas fluorescens; exemption from the requirement of a tolerance. The delta endotoxin of Bacillus thuringiensis variety San Diego encapsulated into killed Pseudomonas fluorescens is...

  15. Carbon beam dosimetry using VIP polymer gel and MRI

    DEFF Research Database (Denmark)

    Kantemiris, I; Petrokokkinos, L; Angelopoulos, A

    2009-01-01

    VIP polymer gel dosimeter was used for Carbon ion beam dosimetry using a 150 MeV/n beam with 10 Gy plateau dose and a SOBP irradiation scheme with 5 Gy Bragg peak dose. The results show a decrease by 8 mm in the expected from Monte Carlo simulation range in water, suggesting that the dosimeter...

  16. Ecology and diversity of Bacillus thuringiensis in soil environment

    African Journals Online (AJOL)

    SERVER

    2007-07-04

    Jul 4, 2007 ... B. thuringiensis isolates were circular, white, flat and undulate or entire. ... India exclusively by the multinational organizations (Sax- ena, 2000). However ... tive, reproductive and crystal structure determination. Physiological.

  17. Evaluation of Genotoxic and Cytotoxic Effects in Human Peripheral Blood Lymphocytes Exposed In Vitro to Neonicotinoid Insecticides News

    Directory of Open Access Journals (Sweden)

    María Elena Calderón-Segura

    2012-01-01

    Full Text Available Calypso (thiacloprid, Poncho (clothianidin, Gaucho (imidacloprid, and Jade (imidacloprid are commercial neonicotinoid insecticides, a new class of agrochemicals in México. However, genotoxic and cytotoxic studies have not been performed. In the present study, human peripheral blood lymphocytes (PBL were exposed in vitro to different concentrations of the four insecticides. The genotoxic and cytotoxic effects were evaluated using the alkaline comet and trypan blue dye exclusion assays. DNA damage was evaluated using two genotoxicity parameters: tail length and comet frequency. Exposure to 9.5×10-6 to 5.7×10-5 M Jade; 2.8×10-4 to 1.7×10-3 M Gaucho; 0.6×10-1 to 1.4×10-1 M Calypso; 1.2×10-1 to 9.5×10-1 M Poncho for 2 h induced a significant increase DNA damage with a concentration-dependent relationship. Jade was the most genotoxic of the four insecticides studied. Cytotoxicity was observed in cells exposed to 18×10-3 M Jade, 2.0×10-3 M Gaucho, 2.0×10-1 M Calypso, 1.07 M Poncho, and cell death occurred at 30×10-3 M Jade, 3.3×10-3 M Gaucho, 2.8×10-1 M Calypso, and 1.42 M Poncho. This study provides the first report of genotoxic and cytotoxic effects in PBL following in vitro exposure to commercial neonicotinoid insecticides.

  18. First Lady Hillary Clinton gets ready to speak to group at VIP viewing site

    Science.gov (United States)

    1999-01-01

    In the VIP Lounge, Apollo/Saturn V Center, First Lady Hillary Rodham Clinton gets ready to speak to the group gathered there before the launch of STS-93. At right is NASA Administrator Daniel Goldin. Liftoff of Space Shuttle Columbia is scheduled for 12:36 a.m. EDT July 20. Much attention has been generated over the launch due to Commander Eileen M. Collins, the first woman to serve as commander of a Shuttle mission. The primary payload of the five-day mission is the Chandra X-ray Observatory, which will allow scientists from around the world to study some of the most distant, powerful and dynamic objects in the universe. The new telescope is 20 to 50 times more sensitive than any previous X- ray telescope and is expected to unlock the secrets of supernovae, quasars and black holes.

  19. Persistence of Toxic Activity of Fermentation Extracts from Bacillus thuringiensis var. israelensis after More Than Three Decades of Storage

    Directory of Open Access Journals (Sweden)

    Luis Jesús Galán-Wong

    2017-01-01

    Full Text Available This study was carried out to determine the persistence of toxicity of fermentation extracts of Bacillus thuringiensis var. israelensis after more than three decades of storage. For this purpose, a population of Aedes aegypti was established. The mortality rate of 20 spore-crystal extracts purified using the acetone-lactose coprecipitation method was measured and evaluated by bioassays according to a modified WHO protocol. The extracts with the highest mortality rate were determined in triplicate by their LD50 and LD98. All extracts showed toxicity at the highest tested dose (1000 ppm and some, such as strains 3260 and 3501, still killed larvae at doses as low as 0.01 ppm. These data are surprising because no study on the activity of B. thuringiensis toxic proteins after such a long storage time has been reported.

  20. Histopathology and the lethal effect of Cry proteins and strains of Bacillus thuringiensis Berliner in Spodoptera frugiperda J.E. Smith Caterpillars (Lepidoptera, Noctuidae

    Directory of Open Access Journals (Sweden)

    N. Knaak

    Full Text Available Among the phytophagous insects which attack crops, the fall armyworm, Spodoptera frugiperda (J.E. Smith, 1797 (Lepidoptera, Noctuidae is particularly harmful in the initial growth phase of rice plants. As a potential means of controlling this pest, and considering that the entomopathogen Bacillus thuringiensis Berliner demonstrates toxicity due to synthesis of the Cry protein, the present study was undertaken to evaluate this toxic effect of B. thuringiensis thuringiensis 407 (pH 408 and B. thuringiensis kurstaki HD-73 on S. frugiperda. The following method was used. Both bacterial strains were evaluated in vitro in 1st instar S. frugiperda caterpillars, by means of histopathological assays. The Cry1Ab and Cry1Ac proteins, codified by the respective strains of B. thuringiensis, were evaluated in vivo by bioassays of 1st instar S. frugiperda caterpillars in order to determine the Mean Lethal Concentration (LC50. The results of the histopathological analysis of the midget of S. frugiperda caterpillars demonstrate that treatment with the B. thuringiensis thuringiensis strain was more efficient, because the degradations of the microvilosities started 9 hours after treatment application (HAT, while in the B. thuringiensis kurstaki the same effect was noticed only after 12 HAT. Toxicity data of the Cry1Ab and Cry1Ac proteins presented for the target-species LC50 levels of 9.29 and 1.79 μg.cm-2 respectively. The strains and proteins synthesised by B. thuringiensis thuringiensis and B. thuringiensis kurstaki are effective in controlling S. frugiperda, and may be used to produce new biopesticides or the genes may be utilised in the genetic transformation of Oryza sativa L.

  1. Toxicidad de δ-endotoxinas recombinantes de Bacillus thuringiensis sobre larvas de la polilla guatemalteca (Tecia solanivora (Lepidóptera: Gelechiidae

    Directory of Open Access Journals (Sweden)

    Javier Hernández-Fernández

    2008-07-01

    Full Text Available Con el objetivo de determinar la actividad tóxica específica de las proteínas recombinantes Cry1Aa, Cry1Ac, Cry1B y Cry1C de Bacillus thuringiensis (Bt, sobre larvas de primer instar de Tecia solanivora se estableció la CL50 para las toxinas. Para este fin se implementó la cría masiva de este insecto bajo condiciones de laboratorio, 58±5% de humedad relativa, 18±5ºC de temperatura y un fotoperiodo de 23 h oscuridad y 1 h luz. Se utilizó una dieta seminatural consistente en láminas de papa variedad parda pastusa autoclavada con solución preservante (ácido ascórbico 7 g/L y metilparabeno 5 g/L, ya que fue estable en el tiempo, garantizó la reproducibilidad de los resultados y fue de fácil evaluación. Las proteínas recombinantes se evaluaron a una concentración de 0,1 μg/cm2. Los resultados obtenidos de porcentaje de mortalidad indicaron que no había diferencias entre las delta-endotoxinas recombinantes Cry1Aa, Cry1Ac, Cry1B y Cry1C de Bt (P

  2. The contribution of agricultural insecticide use to increasing insecticide resistance in African malaria vectors

    OpenAIRE

    Reid, Molly C.; McKenzie, F. Ellis

    2016-01-01

    The fight against malaria is increasingly threatened by failures in vector control due to growing insecticide resistance. This review examines the recent primary research that addresses the putative relationship between agricultural insecticide use and trends in insecticide resistance. To do so, descriptive evidence offered by the new research was categorized, and additional factors that impact the relationship between agricultural insecticide use and observed insecticide resistance in malari...

  3. Diagnostic Doses of Insecticides for Adult Aedes aegypti to Assess Insecticide Resistance in Cuba.

    Science.gov (United States)

    Rodríguez, María Magdalena; Crespo, Ariel; Hurtado, Daymi; Fuentes, Ilario; Rey, Jorge; Bisset, Juan Andrés

    2017-06-01

    The objective of this study was to determine diagnostic doses (DDs) of 5 insecticides for the Rockefeller susceptible strain of Aedes aegypti , using the Centers for Disease Control and Prevention (CDC) bottle bioassay as a tool for monitoring insecticide resistance in the Cuban vector control program. The 30-min DD values determined in this study were 13.5 μg/ml, 6.5 μg/ml, 6 μg/ml, 90.0 μg/ml, and 15.0 μg/ml for cypermethrin, deltamethrin, lambda-cyhalothrin, chlorpyrifos, and propoxur, respectively. To compare the reliability of CDC bottle bioassay with the World Health Organization susceptible test, 3 insecticide-resistant strains were evaluated for deltamethrin and lambda-cyhalothrin. Results showed that the bottles can be used effectively from 21 to 25 days after treatment and reused up to 4 times, depending on the storage time. The CDC bottle bioassay is an effective tool to assess insecticide resistance in field populations of Ae. aegypti in Cuba and can be incorporated into vector management programs using the diagnostic doses determined in this study.

  4. Survival of diverse bacillus thuringiensis strains in gypsy moth (Lepidotera: Lymantriidae) is correlated with urease production

    Science.gov (United States)

    Bacillus thuringiensis is an entomopathogenic bacterium that can kill a variety of pest insects, but seldom causes epizootics because it replicates poorly in insects. By attempting to repeatedly pass lepidopteran-active B. thuringiensis strains through gypsy moth larvae, we found that only those str...

  5. Antagonistic activity of selected strains of Bacillus thuringiensis ...

    African Journals Online (AJOL)

    121, were effective in the reduction of R. solani infection. In addition, GM-23 increased the length of pepper seedlings. These results suggest that the B. thuringiensis strains studied have an excellent potential to be used as bio-control agents of R.

  6. 38 CFR 74.2 - What are the eligibility requirements a concern must meet for VetBiz VIP Verification Program?

    Science.gov (United States)

    2010-07-01

    ... loans or other Federally assisted financing, is eligible for VetBiz VIP Verification. (e) U.S. Small Business Administration (SBA) Protest Decisions. Any firm registered in the VetBiz VIP database that is found to be ineligible due to an SBA protest decision or other negative finding will be immediately...

  7. Efficacy of etanercept in patients with AA amyloidosis secondary to rheumatoid arthritis.

    Science.gov (United States)

    Nakamura, T; Higashi, S; Tomoda, K; Tsukano, M; Baba, S

    2007-01-01

    The efficacy of biological therapies in rheumatoid arthritis (RA) is well known, but their hypothetical benefit in amyloid A (AA) amyloidosis secondary to RA still remains to be considered. We evaluated the efficacy and safety of etanercept in serum amyloid A (SAA) 1.3 allele Japanese patients with AA amyloidosis secondary to RA. Seven RA patients with histologically confirmed AA amyloidosis and renal involvement who were treated with etanercept were enrolled. They all had the SAA1.3 allele, which has been shown to be a risk factor not only for the association of AA amyloidosis but also for a poor prognosis in Japanese RA patients. Efficacy was assessed as a sustained decrease in RA inflammation and an amelioration of renal function. RA inflammation and AA amyloidosis were improved and stabilized after 43.4 +/- 16.5 weeks. At week 20 the number of tender (p = 0.017) and swollen (p = 0.017) joints, and levels of serum C-reactive protein (p = 0.018) and albumin (p = 0.045) had improved. The values for SAA, serum creatinine, calculated creatinine clearance, and proteinuria also ameliorated. No severe adverse events were observed. One patient eventually had to go on hemodialysis but her tolerance of etanercept remained stable. Etanercept can be used safely and effectively in AA amyloidosis secondary to RA with renal involvement, and is of clinical benefit in the short-term, even in patients on hemodialysis. It appears that SAA1.3 allele may be used as a clinical parameter for the introduction of etanercept in Japanese RA with AA amyloidosis.

  8. The cytochrome P450 2AA gene cluster in zebrafish (Danio rerio): Expression of CYP2AA1 and CYP2AA2 and response to phenobarbital-type inducers

    Energy Technology Data Exchange (ETDEWEB)

    Kubota, Akira [Biology Department, Woods Hole Oceanographic Institution, Woods Hole, MA 02543 (United States); Bainy, Afonso C.D. [Biology Department, Woods Hole Oceanographic Institution, Woods Hole, MA 02543 (United States); Departamento de Bioquímica, CCB, Universidade Federal de Santa Catarina, Florianopolis, SC 88040-900 (Brazil); Woodin, Bruce R.; Goldstone, Jared V. [Biology Department, Woods Hole Oceanographic Institution, Woods Hole, MA 02543 (United States); Stegeman, John J., E-mail: jstegeman@whoi.edu [Biology Department, Woods Hole Oceanographic Institution, Woods Hole, MA 02543 (United States)

    2013-10-01

    The cytochrome P450 (CYP) 2 gene family is the largest and most diverse CYP gene family in vertebrates. In zebrafish, we have identified 10 genes in a new subfamily, CYP2AA, which does not show orthology to any human or other mammalian CYP genes. Here we report evolutionary and structural relationships of the 10 CYP2AA genes and expression of the first two genes, CYP2AA1 and CYP2AA2. Parsimony reconstruction of the tandem duplication pattern for the CYP2AA cluster suggests that CYP2AA1, CYP2AA2 and CYP2AA3 likely arose in the earlier duplication events and thus are most diverged in function from the other CYP2AAs. On the other hand, CYP2AA8 and CYP2AA9 are genes that arose in the latest duplication event, implying functional similarity between these two CYPs. A molecular model of CYP2AA1 showing the sequence conservation across the CYP2AA cluster reveals that the regions with the highest variability within the cluster map onto CYP2AA1 near the substrate access channels, suggesting differing substrate specificities. Zebrafish CYP2AA1 transcript was expressed predominantly in the intestine, while CYP2AA2 was most highly expressed in the kidney, suggesting differing roles in physiology. In the liver CYP2AA2 expression but not that of CYP2AA1, was increased by 1,4-bis [2-(3,5-dichloropyridyloxy)] benzene (TCPOBOP) and, to a lesser extent, by phenobarbital (PB). In contrast, pregnenolone 16α-carbonitrile (PCN) increased CYP2AA1 expression, but not CYP2AA2 in the liver. The results identify a CYP2 subfamily in zebrafish that includes genes apparently induced by PB-type chemicals and PXR agonists, the first concrete in vivo evidence for a PB-type response in fish. - Highlights: • A tandemly duplicated cluster of ten CYP2AA genes was described in zebrafish. • Parsimony and duplication analyses suggest pathways to CYP2AA diversity. • Homology models reveal amino acid positions possibly related to functional diversity. • The CYP2AA locus does not share synteny with

  9. The cytochrome P450 2AA gene cluster in zebrafish (Danio rerio): Expression of CYP2AA1 and CYP2AA2 and response to phenobarbital-type inducers

    International Nuclear Information System (INIS)

    Kubota, Akira; Bainy, Afonso C.D.; Woodin, Bruce R.; Goldstone, Jared V.; Stegeman, John J.

    2013-01-01

    The cytochrome P450 (CYP) 2 gene family is the largest and most diverse CYP gene family in vertebrates. In zebrafish, we have identified 10 genes in a new subfamily, CYP2AA, which does not show orthology to any human or other mammalian CYP genes. Here we report evolutionary and structural relationships of the 10 CYP2AA genes and expression of the first two genes, CYP2AA1 and CYP2AA2. Parsimony reconstruction of the tandem duplication pattern for the CYP2AA cluster suggests that CYP2AA1, CYP2AA2 and CYP2AA3 likely arose in the earlier duplication events and thus are most diverged in function from the other CYP2AAs. On the other hand, CYP2AA8 and CYP2AA9 are genes that arose in the latest duplication event, implying functional similarity between these two CYPs. A molecular model of CYP2AA1 showing the sequence conservation across the CYP2AA cluster reveals that the regions with the highest variability within the cluster map onto CYP2AA1 near the substrate access channels, suggesting differing substrate specificities. Zebrafish CYP2AA1 transcript was expressed predominantly in the intestine, while CYP2AA2 was most highly expressed in the kidney, suggesting differing roles in physiology. In the liver CYP2AA2 expression but not that of CYP2AA1, was increased by 1,4-bis [2-(3,5-dichloropyridyloxy)] benzene (TCPOBOP) and, to a lesser extent, by phenobarbital (PB). In contrast, pregnenolone 16α-carbonitrile (PCN) increased CYP2AA1 expression, but not CYP2AA2 in the liver. The results identify a CYP2 subfamily in zebrafish that includes genes apparently induced by PB-type chemicals and PXR agonists, the first concrete in vivo evidence for a PB-type response in fish. - Highlights: • A tandemly duplicated cluster of ten CYP2AA genes was described in zebrafish. • Parsimony and duplication analyses suggest pathways to CYP2AA diversity. • Homology models reveal amino acid positions possibly related to functional diversity. • The CYP2AA locus does not share synteny with

  10. Effect of chemical additives on Bacillus thuringiensis (Bacillales: Bacillaceae) against Plutella xylostella (Lepidoptera: Pyralidae)

    Czech Academy of Sciences Publication Activity Database

    Zhang, L.; Qiu, S.; Huang, T.; Huang, Z.; Xu, L.; Wu, C.; Gelbič, Ivan; Guan, X.

    2013-01-01

    Roč. 106, č. 3 (2013), s. 1075-1080 ISSN 0022-0493 Institutional research plan: CEZ:AV0Z50070508 Keywords : additives * Bacillus thuringiensis * biocontrol Subject RIV: GF - Plant Pathology, Vermin, Weed, Plant Protection Impact factor: 1.605, year: 2013 http://www.bioone.org/doi/pdf/10.1603/EC12288

  11. Synthesis and biological activity of a new class of insecticides: the N-(5-aryl-1,3,4-thiadiazol-2-yl)amides.

    Science.gov (United States)

    Eckelbarger, Joseph D; Parker, Marshall H; Yap, Maurice Ch; Buysse, Ann M; Babcock, Jonathan M; Hunter, Ricky; Adelfinskaya, Yelena; Samaritoni, Jack G; Garizi, Negar; Trullinger, Tony K

    2017-04-01

    Optimization studies on a high-throughput screening (HTS) hit led to the discovery of a series of N-(6-arylpyridazin-3-yl)amides with insecticidal activity. It was hypothesized that the isosteric replacement of the pyridazine ring with a 1,3,4-thiadiazole ring could lead to more potent biological activity and/or a broader sap-feeding pest spectrum. The resulting N-(5-aryl-1,3,4-thiadiazol-2-yl)amides were explored as a new class of insecticides. Several methods for 2-amino-1,3,4-thiadiazole synthesis were used for the preparation of key synthetic intermediates. Subsequent coupling to variously substituted carboxylic acid building blocks furnished the final targets, which were tested for insecticidal activity against susceptible strains of Aphis gossypii (Glover) (cotton aphid), Myzus persicae (Sulzer) (green peach aphid) and Bemisia tabaci (Gennadius) (sweetpotato whitefly). Structure-activity relationship (SAR) studies on both the amide tail and the aryl A-ring of novel N-(5-aryl-1,3,4-thiadiazol-2-yl)amides led to a new class of insecticidal molecules active against sap-feeding insect pests. © 2016 Society of Chemical Industry. © 2016 Society of Chemical Industry.

  12. New experimental limit on Pauli exclusion principle violation by electrons (VIP experiment)

    Energy Technology Data Exchange (ETDEWEB)

    Bartalucci, S [NFN, Laboratori Nazionali di Prascati, C.P. 13, Via E. Fermi 40, I-00044, Frascati (Italy); Bertolucci, S [NFN, Laboratori Nazionali di Prascati, C.P. 13, Via E. Fermi 40, I-00044, Frascati (Italy); Bragadireanu, M [NFN, Laboratori Nazionali di Prascati, C.P. 13, Via E. Fermi 40, I-00044, Frascati (Italy)] (and others)

    2007-05-15

    The Pauli exclusion principle (PEP) represents one of the basic principles of modern physics and, even if there are no compelling reasons to doubt its validity, it still spurs a lively debate, because an intuitive, elementary explanation is still missing, and because of its unique stand among the basic symmetries of physics. A new limit on the probability that PEP is violated by electrons was estabilished by the VIP (Violation of the Pauli exclusion principle) Collaboration, using the method of searching for PEP forbidden atomic transitions in copper. The preliminary value, 1/2{beta}{sup 2} < 4.5 x 10{sup -28}, represents an improvement of about two orders of magnitude of the previous limit. The goal of VIP is to push this limit at the level of 10{sup -30}.

  13. VIP Data Explorer: A Tool for Exploring 30 years of Vegetation Index and Phenology Observations

    Science.gov (United States)

    Barreto-munoz, A.; Didan, K.; Rivera-Camacho, J.; Yitayew, M.; Miura, T.; Tsend-Ayush, J.

    2011-12-01

    Continuous acquisition of global satellite imagery over the years has contributed to the creation of long term data records from AVHRR, MODIS, TM, SPOT-VGT and other sensors. These records account for 30+ years, as these archives grow, they become invaluable tools for environmental, resources management, and climate studies dealing with trends and changes from local, regional to global scale. In this project, the Vegetation Index and Phenology Lab (VIPLab) is processing 30 years of daily global surface reflectance data into an Earth Science Data Record of Vegetation Index and Phenology metrics. Data from AVHRR (N07,N09,N11 and N14) and MODIS (AQUA and TERRA collection 5) for the periods 1981-1999 and 2000-2010, at CMG resolution were processed into one seamless and sensor independent data record using various filtering, continuity and gap filling techniques (Tsend-Ayush et al., AGU 2011, Rivera-Camacho et al, AGU 2011). An interactive online tool (VIP Data Explorer) was developed to support the visualization, qualitative and quantitative exploration, distribution, and documentation of these records using a simple web 2.0 interface. The VIP Data explorer (http://vip.arizona.edu/viplab_data_explorer) can display any combination of multi temporal and multi source data, enable the quickly exploration and cross comparison of the various levels of processing of this data. It uses the Google Earth (GE) model and was developed using the GE API for images rendering, manipulation and geolocation. These ESDRs records can be quickly animated in this environment and explored for visual trends and anomalies detection. Additionally the tool enables extracting and visualizing any land pixel time series while showing the different levels of processing it went through. User can explore this ESDR database within this data explorer GUI environment, and any desired data can be placed into a dynamic "cart" to be ordered and downloaded later. More functionalities are planned and will be

  14. Rice Production without Insecticide in Smallholder Farmer's Field

    Directory of Open Access Journals (Sweden)

    M. P. Ali

    2017-05-01

    Full Text Available Highlights:Use of perching, sweeping, and need based insecticide (IPM technique useage produce at par yields compared to prophylactic insecticide useage in rice fields.There exists a technique that can reduce 75% of insecticide useage in rice field.The results were obtained in cooperation between smallholder rice farmers and researchers of Bangladesh.Currently rice protection from insect pests solely depends on chemical pesticides which have tremendous impact on biodiversity, environment, animal, and human health. To reduce their impact from our society we need to cut pesticide use from agricultural practices. To address this issue, we did an experiment to identify realistic solutions that could help farmers build sustainable crop protection systems and minimize useage of insecticides and thus reduce the impact of pesticides in the environment. Innovations developed jointly by farmers and researchers and evaluated for their potential to be adopted by more farmers. In this paper we tested four management practices jointly with smallholder farmer fields in order to select the best one. Four management practices were used namely, T1 = Prophylactic use of insecticide where insecticide was applied in rice field at every 15 days interval without judging the infestation level; T2 = Perching (that is, placing roosting (perching sites for insectivorous birds within the rice field and concurrent sweep net samples along with need-based insecticide application; T3 = Perching only; and T4 = Farmer's own practices. The results revealed that routine application of insecticides for crop protection is not mandatory which is commonly found at use in rice farmers. In our experiment, where prophylactic method or farmers used 3–4 times insecticides without judging the insect pests infestation level, the similar pest population was found when compared to the field where insecticide was not applied. Our management system reduced by 75% the use of insecticides even

  15. The contribution of agricultural insecticide use to increasing insecticide resistance in African malaria vectors.

    Science.gov (United States)

    Reid, Molly C; McKenzie, F Ellis

    2016-02-19

    The fight against malaria is increasingly threatened by failures in vector control due to growing insecticide resistance. This review examines the recent primary research that addresses the putative relationship between agricultural insecticide use and trends in insecticide resistance. To do so, descriptive evidence offered by the new research was categorized, and additional factors that impact the relationship between agricultural insecticide use and observed insecticide resistance in malaria vectors were identified. In 23 of the 25 relevant recent publications from across Africa, higher resistance in mosquito populations was associated with agricultural insecticide use. This association appears to be affected by crop type, farm pest management strategy and urban development.

  16. Peripheral axotomy of the rat mandibular trigeminal nerve leads to an increase in VIP and decrease of other primary afferent neuropeptides in the spinal trigeminal nucleus.

    Science.gov (United States)

    Atkinson, M E; Shehab, S A

    1986-12-01

    In the vasoactive intestinal polypeptide (VIP)-rich lumbosacral spinal cord, VIP increases at the expense of other neuropeptides after primary sensory nerve axotomy. This study was undertaken to ascertain whether similar changes occur in peripherally axotomised cranial sensory nerves. VIP immunoreactivity increased in the terminal region of the mandibular nerve in the trigeminal nucleus caudalis following unilateral section of the sensory root of the mandibular trigeminal nerve at the foramen orale. Other primary afferent neuropeptides (substance P, cholecystokinin and somatostatin) were depleted and fluoride-resistant acid phosphatase activity was abolished in the same circumscribed areas of the nucleus caudalis. The rise in VIP and depletion of other markers began 4 days postoperatively and was maximal by 10 days, these levels remaining unchanged up to 1 year postoperatively. VIP-immunoreactive cell bodies were absent from trigeminal ganglia from the unoperated side but small and medium cells stained intensely in the ganglia of the operated side after axotomy. These observations indicate that increase of VIP in sensory nerve terminals is a general phenomenon occurring in both cranial and spinal sensory terminal areas. The intense VIP immunoreactivity in axotomised trigeminal ganglia suggests that the increased levels of VIP in the nucleus caudalis are of peripheral origin, indicating a change in expression of neuropeptides within primary afferent neurons following peripheral axotomy.

  17. Isolation and toxicity test of Bacillus thuringiensis from Sekayu region soil, South Sumatra on Spodopteralitura

    Science.gov (United States)

    Afriani, S. R.; Pujiastuti, Y.; Irsan, C.; Damiri, N.; Nugraha, S.; Sembiring, E. R.

    2018-01-01

    This study aimed to obtain bacterial isolates B. thuringiensis potential as a biological control against pests Spodoptera litura. The research was conducted at the Laboratory of Pest and Disease Department, Agricultural Faculty of Sriwijaya University, Campus Inderalaya Ogan Ilir, South Sumatera, from March to June 2017. The study was conducted with survey method and laboratory trial. The results showed that of the 50 soil samples from three villages selected through morphological observation, reaction staining, KOH test, catalase test, producing 13 bacterial isolates. Screening of the 13th toxicity of the isolates suspected B. thuringiensis against S. litura larvae was investigated. Based on the toxicity screening test the following facts were obtained: five isolates ie KJ2M2, KJ3E1, KJ3JB1, KJ3D3 and KJ3D5 were lower toxicity than Dipel, two isolates ie KJ3K4 and KJ3D3 which had the same toxicity to Dipel. Five isolates the KJ3E3, KJ3BW5, KJ3JB5, KJ3D1 and LC2, LC3 known to have effectiveness until the seventh day reached 40%. There was one isolate that is KJ3BW5 which was more effective compared to Dipel as comparison.

  18. A short history of insecticides

    Directory of Open Access Journals (Sweden)

    Oberemok Volodymyr Volodymyrovych

    2015-07-01

    Full Text Available This review contains a brief history of the use of insecticides. The peculiarities, main advantages, and disadvantages of some modern insecticides are described. The names of the discoverers of some of the most popular insecticide preparations on the world market, are listed. The tendencies to find new insecticides to control the quantity of phytophagous insects are discussed. Special attention is paid to the perspective of creating preparations based on nucleic acids, in particular DNA insecticides. The use of insect-specific, short single-stranded DNA fragments as DNA insecticides, is paving the way in the field of “intellectual” insecticides that “think” before they act. It is worth noting, though, that in the near future, the quantity of produced insecticides will increase due to the challenges associated with food production for a rapidly growing population. It is concluded, that an agreeable interaction of scientists and manufacturers of insecticides should lead to the selection of the most optimal solutions for insect pest control, which would be safe, affordable, and effective at the same time.

  19. Expression of the sigma35 and cry2AB genes involved in Bacillus thuringiensis virulence Expressão dos genes sigma35 e cry2AB envolvidos na virulência de Bacillus thuringiensis

    Directory of Open Access Journals (Sweden)

    Ana Maria Guidelli-Thuler

    2009-06-01

    Full Text Available There are several genes involved in Bacillus thuringiensis sporulation. The regulation and expression of these genes results in an upregulation in Cry protein production, and this is responsible for the death of insect larvae infected by Bacillus thuringiensis. Gene expression was monitored in Bacillus thuringiensis during three developmental phases. DNA macroarrays were constructed for selected genes whose sequences are available in the GenBank database. These genes were hybridized to cDNA sequences from B. thuringiensis var. kurstaki HD-1. cDNA probes were synthesized by reverse transcription from B. thuringiensis RNA templates extracted during the exponential (log growth, stationary and sporulation phases, and labeled with 33PadCTP. Two genes were differentially expressed levels during the different developmental phases. One of these genes is related to sigma factor (sigma35, and the other is a cry gene (cry2Ab. There were differences between the differential levels of expression of various genes and among the expression detected for different combinations of the sigma factor and cry2Ab genes. The maximum difference in expression was observed for the gene encoding sigma35 factor in the log phase, which was also expressed at a high level during the sporulation phase. The cry2Ab gene was only expressed at a high level in the log phase, but at very low levels in the other phases when compared to the sigma35.Muitos genes estão envolvidos nos mecanismos de esporulação da bactéria Bacillus thuringiensis. A regulação e expressão desses genes resultam em uma produção massiva da proteína Cry, responsável pela morte das larvas de muitos insetos. Neste trabalho monitorou-se a expressão de genes de Bacillus thuringiensis, ao longo de três fases de seu desenvolvimento. Foram construídos macroarrays de DNA dos genes selecionados, cujas seqüências estão disponibilizadas no GenBank. Estes genes foram hibridizados com cDNAs obtidos de B

  20. Susceptibility of Spodoptera littoralis caterpillars to entomopathogenic nematode Steinernema feltiae after consumption of non-specific transgenic plants

    Czech Academy of Sciences Publication Activity Database

    Adel, M. M.; Hussein, Hany; Habuštová, Oxana; Sehnal, František

    2012-01-01

    Roč. 48, č. 18 (2012), s. 2251-2260 ISSN 0323-5408 R&D Projects: GA MŠk 1M06030 Institutional research plan: CEZ:AV0Z50070508 Keywords : Steinernema feltiae * Bacillus thuringiensis * Cry3Aa Subject RIV: ED - Physiology

  1. Composition of the Essential Oil of Salvia ballotiflora (Lamiaceae and Its Insecticidal Activity

    Directory of Open Access Journals (Sweden)

    Norma Cecilia Cárdenas-Ortega

    2015-05-01

    Full Text Available Essential oils can be used as an alternative to using synthetic insecticides for pest management. Therefore, the insectistatic and insecticidal activities of the essential oil of aerial parts of Salvia ballotiflora (Lamiaceae were tested against the fall armyworm Spodoptera frugiperda (Lepidoptera: Noctuidae. The results demonstrated insecticidal and insectistatical activities against this insect pest with concentrations at 80 µg·mL−1 resulting in 20% larval viability and 10% pupal viability. The larval viability fifty (LV50 corresponded to a concentration of 128.8 µg·mL−1. This oil also increased the duration of the larval phase by 5.5 days and reduced the pupal weight by 29.2% withrespect to the control. The GC-MS analysis of the essential oil of S. ballotiflora showed its main components to be caryophyllene oxide (15.97%, and β-caryophyllene (12.74%, which showed insecticidal and insectistatical activities against S. frugiperda. The insecticidal activity of β-caryophyllene began at 80 µg·mL−1, giving a larval viability of 25% and viability pupal of 20%. The insectistatic activity also started at 80 µg·mL−1 reducing the pupal weight by 22.1% with respect to control. Caryophyllene oxide showed insecticidal activity at 80 µg·mL−1 giving a larval viability of 35% and viability pupal of 20%.The insectistatic activity started at 400 µg·mL−1 and increased the larval phase by 8.8% days with respect to control. The LV50 values for these compounds were 153.1 and 146.5 µg·mL−1, respectively.

  2. Design, Synthesis and Insecticidal Activity of Novel Phenylurea Derivatives

    Directory of Open Access Journals (Sweden)

    Jialong Sun

    2015-03-01

    Full Text Available A series of novel phenylurea derivatives were designed and synthesized according to the method of active groups linkage and the principle of aromatic groups bioisosterism in this study. The structures of the novel phenylurea derivatives were confirmed based on ESI-MS, IR and 1H-NMR spectral data. All of the compounds were evaluated for the insecticidal activity against the third instars larvae of Spodoptera exigua Hiibner, Plutella xyllostella Linnaeus, Helicoverpa armigera Hubner and Pieris rapae Linne respectively, at the concentration of 10 mg/L. The results showed that all of the derivatives displayed strong insecticidal activity. Most of the compounds presented higher insecticidal activity against S. exigua than the reference compounds tebufenozide, chlorbenzuron and metaflumizone. Among the synthesized compounds, 3b, 3d, 3f, 4b and 4g displayed broad spectrum insecticidal activity.

  3. 40 CFR 180.1011 - Viable spores of the microorganism Bacillus thuringiensis Berliner; exemption from the...

    Science.gov (United States)

    2010-07-01

    ... EXEMPTIONS FOR PESTICIDE CHEMICAL RESIDUES IN FOOD Exemptions From Tolerances § 180.1011 Viable spores of the... characteristics of the parent strain or contamination by other microorganisms. (3) Each lot of spore preparation... production is a Bacillus thuringiensis strain which does not produce β-exotoxin under standard manufacturing...

  4. Rapidity and energy dependence of the electric charge correlations in A+A collisions from 20A to 158A GeV

    NARCIS (Netherlands)

    Botje, M.A.J.; van Leeuwen, M.; Mischke, A.

    2007-01-01

    Results from electric charge correlations studied with the Balance Function (BF) method in A+A collisions from 20A to 158A GeV are presented in two different rapidity intervals: In the midrapidity region we observe a decrease of the width of the BF distribution with increasing centrality of the

  5. Impacts of Bacillus thuringiensis var. israelensis and Bacillus ...

    African Journals Online (AJOL)

    The study assessed the impact of bio-larvicides- Bacillus thuringiensis var. israelensis (Bti) and B. sphaericus (Bs) on anopheline mosquito larval densities in four selected areas of Lusaka urban district. Larval densities were determined using a standard WHO protocol at each study area prior to and after larviciding.

  6. pH-controlled Bacillus thuringiensis Cry1Ac protoxin loading and release from polyelectrolyte microcapsules.

    Directory of Open Access Journals (Sweden)

    Wenhui Yang

    Full Text Available Crystal proteins synthesized by Bacillus thuringiensis (Bt have been used as biopesticides because of their toxicity to the insect larval hosts. To protect the proteins from environmental stress to extend their activity, we have developed a new microcapsule formulation. Poly (acrylic acid (PAH and poly (styrene sulfonate (PSS were fabricated through layer-by-layer self-assembly based on a CaCO(3 core. Cry1Ac protoxins were loaded into microcapsules through layer-by-layer self-assembly at low pH, and the encapsulated product was stored in water at 4°C. Scanning electron microscopy (SEM was used to observe the morphology of the capsules. To confirm the successful encapsulation, the loading results were observed with a confocal laser scattering microscope (CLSM, using fluorescein-labeled Cry1Ac protoxin (FITC-Cry1Ac. The protoxins were released from the capsule under the alkaline condition corresponding to the midgut of certain insects, a condition which seldom exists elsewhere in the environment. The following bioassay experiment demonstrated that the microcapsules with Cry1Ac protoxins displayed approximately equivalent insecticidal activity to the Asian corn borer compared with free Cry1Ac protoxins, and empty capsules proved to have no effect on insects. Further result also indicated that the formulation could keep stable under the condition of heat and desiccation. These results suggest that this formulation provides a promising methodology that protects protoxins from the environment and releases them specifically in the target insects' midgut, which has shown potential as biopesticide in the field.

  7. Study of the effect of cerium nitrate on AA2024-T3 by means of electrochemical micro-cell technique

    International Nuclear Information System (INIS)

    Paussa, L.; Andreatta, F.; Rosero Navarro, N.C.; Durán, A.; Fedrizzi, L.

    2012-01-01

    Highlights: ► We evaluate the cerium nitrate effect on the electrochemical behavior of AA2024-T3. ► We examine how AA2024-T3 microstructure affects cerium precipitation mechanism. ► The entire AA2024-T3 surface is involved in cerium precipitation. ► Anodic and cathodic inhibitions are both provided by cerium precipitation. ► Mg-rich intermetallics are preferential sites for cerium precipitation. - Abstract: This work evaluates the effect of cerium nitrate as corrosion inhibitor for AA2024-T3 in the view of its introduction in sol–gel coatings able to provide self-healing ability. Since it is well established that deposition of Ce species is activated by the local pH increase, the objective of this paper is to investigate the behavior of AA2024-T3 (open circuit potential and polarization curves) in the presence of Ce species in aggressive solutions by means of a local technique, the electrochemical micro-cell. This technique enables the investigation of small areas with resolution in the micrometer range by the use of glass capillaries to define the working electrode area. The micro-cell results clearly displayed that the entire AA2024-T3 area exposed to the cerium-containing electrolyte was involved in the cerium precipitation mechanism. The heterogeneous electrochemical behavior of the microstructure is minimized by the formation of a cerium-containing layer able to protect the metal substrate.

  8. Operational Evaluation Of Vectomax® WSP (Bacillus thuringiensis Subsp. israelensis+Bacillus sphaericus) Against Larval Culex pipiens in Septic Tanks (1).

    Science.gov (United States)

    Cetin, Huseyin; Oz, Emre; Yanikoglu, Atila; Cilek, James E

    2015-06-01

    The residual effectiveness of VectoMax® WSP (a water-soluble pouch formulation containing a combination of Bacillus thuringiensis subsp. israelensis strain AM65-52 and B. sphaericus strain ABTS 1743) when applied to septic tanks against 3rd- and 4th-stage larvae of Culex pipiens L. was evaluated in this study. This formulation was evaluated at operational application rates of 1 pouch (10 g) and 2 pouches (20 g) per septic tank. Both application rates resulted in >96% control of larvae for 24 days. Operationally, VectoMax WSP has proven to be a useful tool for the nonchemical control of Culex species in septic tank environments.

  9. Produccion por tecnologia de fermentacion de bacillus thuringiensis utilizando medios alternativos

    Directory of Open Access Journals (Sweden)

    Yaneth Amparo Muñoz-Peñalosa

    2000-01-01

    Full Text Available In the production by fermentation technology Bacillus thuringiensis of five alternative methods they were studied. The results of cell growth, working-level 100ml in static culture and temperature of 28 ° C, mostraronque the optimal substrate corresponded to the environment in which molasses and rice powder was added (alternative Medium No. 1] The cell development using 100 ml of substrate was studied with reciprocating shaking 110 rpm. in this test was determined filter curve creciemiento medio.The inoculum, allowed tiempode set the process in 6 hours. For the development of fermentations, are counted with the experimental equipment, glass bioreactor in two liters of capacity and aeration devices, mechanical stirring, sampling and output gases.The fermentation in the production of Bacillus thuringiensis is the type discontinuous submerged aerobic process and growth into account .Teniendo bibliographic information and preliminary results of the study, fixed working parameters were determined for production by fermentation of Bacillus thuringiensis, being alternative means No. 1, volume 1 liter temperature 28 ° C and cell concentration of the inoculum. To determine the optimum parameters of fermantacion was used a factorial design of experiments of the type 22, (two variables at two levels, with aeration (3.2-0.5 VVM and agitation (110-210 rpm .The fermentations performed 7, 4 design and 3 the average level of the variables. For monitoring fermentation sample was taken every 12 hours and cell concentration (Chamber of Neuvauer and pH was analyzed. The results of cell concentration measurement for fermentations at 60 hours shows that optimum working conditions and limitations correspond to the values ​​of the variable, 3.2 VVM aeration and agitation 210 Variable rpm.Significant was the aeration of pH in the fermentation media change neutral to acid and ended as a staple. A fermentations I were efectuo controlde microbiological quality, Gram

  10. Effects of chronic ethanol ingestion on arachidonic acid (AA) metabolism in rat macrophages

    International Nuclear Information System (INIS)

    Chanmugam, P.; Boudreau, M.; Hymel, G.; Jeffers, G.; Hwang, D.H.

    1986-01-01

    In a previous report, preincubation of rat platelets with ethanol resulted in dose dependent inhibition of AA metabolites whereas chronic ingestion of ethanol enhanced the synthesis of AA metabolites. Thus, the authors studied whether chronic ethanol ingestion also affects AA metabolism in MACS. Two groups of rats (10 each) were fed DeCarli/Lieber liquid diet containing 36 caloric % ethanol for 3 weeks. The control group was pair fed the same diet made isocaloric with dextrin-maltose. Resident MACS were collected by peritoneal lavage. The monolayers of MACS were incubated for 20 min with calcium ionophore (5μg/ml), and the incubation stopped with 4 vol. of ethanol. PGE 2 , LTB 4 and 5-HETE were assayed by radioimmunoassay. The results indicated that chronic ethanol ingestion did not affect the capacity of MACS to synthesize AA metabolites. There was also no difference in the levels of AA metabolites in heart and lung homogenates between the two groups

  11. Pathogenicity of Bacillus thuringiensis isolated from two species of Acromyrmex (Hymenoptera, Formicidae

    Directory of Open Access Journals (Sweden)

    L. M. N. Pinto

    Full Text Available The control of Acromyrmex leaf-cutting ants is necessary due to the severe damage they cause to diverse crops. A possibility was to control them using the bacterium Bacillus thuringiensis (Bt that characteristically produces insecticidal crystal proteins (ICPs. The ICPs have been effective in controlling lepidopterans, dipterans, and coleopterans, but their action against hymenopterans is unknown. This paper describes an attempt to isolate Bt from ants of two Acromyrmex species, to evaluate its pathogenicity towards these ants, and to test isolates by PCR. Bacterial isolates of Bt obtained from A. crassispinus and A. lundi have been assayed against A. lundi in the laboratory. The bioassays were carried out in BOD at 25°C, with a 12-hour photoperiod, until the seventh day after treatment. The Bt isolates obtained were submitted to total DNA extraction and tested by PCR with primers specific to cry genes. The results showed Bt presence in 40% of the assessed samples. The data from the in vivo assays showed a mortality rate higher than 50% in the target population, with the Bt HA48 isolate causing 100% of corrected mortality. The PCR results of Bt isolates showed a magnification of DNA fragments relative to cry1 genes in 22% of the isolates, and cry9 in 67%. Cry2, cry3, cry7, and cry8 genes were not detected in the tested samples, and 22% had no magnified DNA fragments corresponding to the assessed cry genes. The results are promising not only regarding allele identification in new isolates, but also fort the assays aimed at determining the Bt HA48 LC50's, which can eventually be applied in controlling of Acromyrmex leaf-cutting ants.

  12. Isolation and characterization of radioresistant mutants in Bacillus subtilis and Bacillus thuringiensis

    International Nuclear Information System (INIS)

    Kalinin, V.L.; Petrov, V.N.; Petrova, T.M.

    1981-01-01

    Vegetative cells of Bac. thuringiensis var. galleriae (the wild-type strain 351) are much more sensitive to lethal effects of UV light and 60 Co-γ-rays than those of Bac. subtilis (the wild-type strain 168). This difference is less pronounced for spores of these strains. By means of repeated γ-irradiation-regrowth cycles radioresistant mutants Bac. thuringiensis Gamsup(r) 14 and Bac. subtilis Gamsup(r) 9 were selected. The vegetative cells of these mutants are correspondingly 19 times and 3.9 times more resistant to lethal effects of γ-radiation than the cells of the parental strains. The resistance of the Gamsup(r) mutant cells to lethal effects of UV light and H 2 O 2 is also increased. The spores of the Gamsup(r) 14 mutant are 1.5-1.7 times more resistant to γ-radiation and UV light than the wild-type spores. The radioresistant mutants and the parental strains do not vary in their capacity for host-cell reactivation of UV- or γ-irradiated phages Tg13 and 105

  13. Extrinsic control of the release of galanin and VIP from intrinsic nerves of isolated, perfused, porcine ileum

    DEFF Research Database (Denmark)

    Messell, T; Harling, H; Poulsen, Steen Seier

    1992-01-01

    By immunohistochemistry galanin-like immunoreactivity and vasoactive intestinal polypeptide (VIP)-like immunoreactivity were found in nerve cell bodies mostly in the submucous plexus and in nerve fibres in the mucosa, submucosa and muscularis including the myenteric plexus of the porcine ileum an...... was not influenced by atropine. Our results suggest that the galanin- and VIP-producing intrinsic neurons receive inhibitory signals by noradrenergic nerve fibers and stimulatory signals mediated by cholinergic nerves, possibly via a cholinergic interneuron....

  14. Venus Interior Probe Using In-situ Power and Propulsion (VIP-INSPR), Phase I

    Data.gov (United States)

    National Aeronautics and Space Administration — We envision a novel architecture for Venus Interior Probes based on in-situ resources for power generation (VIP-INSPR). Proposed Venus probe is based on the...

  15. Effect of Bacillus thuringiensis parasporal toxin on stimulating of IL-2 and IL-5 cytokines production

    Directory of Open Access Journals (Sweden)

    Marzieh Soleimany

    2018-03-01

    Full Text Available Introduction:Bacillus thuringiensis, is a Gram-positive spore-forming bacterium that produces crystalline parasporal protein (Cry during sporulation. Some of these Cry toxins do not show cytotoxicity against insects but they are capable to kill some human and animal cancer cells. The aim of this study was to verify whether cytocidal parasporal of B thuringiensis strains have immunostimulatory activity on human peripheral blood mononuclear cells (PBMNC and to evaluate the ability of IL-2 and IL-5 production. Materials and methods: B. thuringiensis toxin with cytocidal activity was isolated and treated with proteinase K. PBMNC was cultured and treated with activated crystal proteins. We evaluated the ability of different cytokines production with Flow Cytometry. Results: In this study, immune stimulatory toxins Cry1 were distinguished. This toxin can stimulate production of cytokines IL-2 and stop production of IL-5. Discussion and conclusion: According to anti-cancer effect of B. thuringiensis toxins and also immune stimulatory effect, with more research these toxins can be introduced as immunotherapy drug in cancer treatment.

  16. Decrease of insecticide resistance over generations without exposure to insecticides in Nilaparvata lugens (Hemipteran: Delphacidae).

    Science.gov (United States)

    Yang, Yajun; Dong, Biqin; Xu, Hongxing; Zheng, Xusong; Tian, Junce; Heong, Kongleun; Lu, Zhongxian

    2014-08-01

    The brown planthopper, Nilaparvata lugens (Stål), is one of the most important insect pests on paddy rice in tropical and temperate Asia. Overuse and misuse of insecticides have resulted in the development of high resistance to many different insecticides in this pest. Studies were conducted to evaluate the change of resistance level to four insecticides over 15 generations without any exposure to insecticides in brown planthopper. After 15 generations' rearing without exposure to insecticide, brown planthopper could reverse the resistance to imidacloprid, chlorpyrifos, fipronil, and fenobucarb. The range and style of resistance reversal of brown planthopper differed when treated with four different insecticides. To monitor potential changes in insect physiological responses, we measured the activity of each of the three selected enzymes, including acetylcholinesterases (AChE), general esterases (EST), and glutathione S-transferases. After multiple generations' rearing without exposure to insecticide, AChE and EST activities of brown planthopper declined with the increased generations, suggesting that the brown planthopper population adjusted activities of EST and AChE to adapt to the non-insecticide environment. These findings suggest that the reducing, temporary stop, or rotation of insecticide application could be incorporated into the brown planthopper management.

  17. Involvement of microRNA miR-2b-3p in regulation of metabolic resistance to insecticides in Plutella xylostella.

    Science.gov (United States)

    Etebari, K; Afrad, M H; Tang, B; Silva, R; Furlong, M J; Asgari, S

    2018-03-24

    The diamondback moth, Plutella xylostella, has developed extremely high levels of resistance to chlorantraniliprole and other classes of insecticides in the field. As microRNAs (miRNAs) play important roles in various biological processes through gene regulation, we examined the miRNA profile of P. xylostella in response to chlorantraniliprole exposure. RNA sequencing analysis showed that insecticide treatment caused significant changes in the abundance of some miRNAs. Increasing exposure time and insecticide concentration induced more dysregulated miRNAs in P. xylostella larvae. We also screened potential target genes for some of the differentially expressed miRNAs (such as miR-2b-3p, miR-14b-5p and let-7-5p), which may play important roles in insecticide resistance development. Exposure of P. xylostella larvae to chlorantraniliprole caused considerable overexpression in the transcript levels of potential target genes cytochrome P450 9f2 (CYP9F2) and 307a1 (CYP307a1). Application of miR-2b-3p and miR-14b-5p mimics significantly suppressed the relative transcript levels of CYP9F2 and CYP307a1, respectively, in a P. xylostella cell line. Furthermore, enrichment of P. xylostella diet with miR-2b-3p mimics significantly increased mortality in deltamethrin-resistant larvae when exposed to deltamethrin. The results suggest that miR-2b-3p may suppress CYP9F2 transcript levels in P. xylostella and consequently inhibit larval detoxification pathways. The findings provide an insight into possible role of miRNAs in regulation of metabolic resistance of insects to insecticides. © 2018 The Royal Entomological Society.

  18. Anodic galvanostatic polarization of AA2024-T3 aircraft alloy in conventional mineral acids

    Energy Technology Data Exchange (ETDEWEB)

    Kozhukharov, S., E-mail: stephko1980@abv.bg [Department of Chemical Sciences, University of Chemical Technology and Metallurgy, 8 “Kliment Okhridski” Blvd, 1756, Sofia (Bulgaria); Girginov, Ch. [Department of Chemical Sciences, University of Chemical Technology and Metallurgy, 8 “Kliment Okhridski” Blvd, 1756, Sofia (Bulgaria); Avramova, I. [Institute of General and Inorganic Chemistry, Bulgarian Academy of Science, 11 “Georgi Bonchev” Str., 1113, Sofia (Bulgaria); Machkova, M. [Department of Chemical Sciences, University of Chemical Technology and Metallurgy, 8 “Kliment Okhridski” Blvd, 1756, Sofia (Bulgaria)

    2016-09-01

    The present study is devoted to the determination of the impact of the anodization of AA2024-T3 alloys in HCl, HNO{sub 3}, H{sub 2}SO{sub 4} or H{sub 3}PO{sub 4} on the samples’ surface morphology and properties. Subsequent systematic assessments were performed by Scanning Electron Microscopy (SEM), Energy Dispersion X-Ray Spectroscopy (EDX) and X-ray Photoelectron Spectroscopy (XPS). These observations were combined with Linear Voltammetry (LVA) and Electrochemical Impedance Spectroscopy (EIS) after 48 and 168 h of exposure to a 3.5% NaCl model corrosive medium. The main result is, that completely different effects were observed in accordance to the acid used. It was established that the monoprotonic acids have a deep destructive effect due to dissolution of the alloy components, whereas the polyprotonic ones possess either indistinguishable influence, or surface film formation. - Highlights: • AA2024 was polarized anodically in 15%{sub wt} acid solutions at 15 mA cm{sup −2} for 2 h. • Four mineral acids were selected for investigation: HCl, HNO{sub 3}, H{sub 2}SO{sub 4} and H{sub 3}PO{sub 4}. • SEM, EDX and XPS were applied for morphological description. • Electrochemical characterizations were performed by EIS and linear voltammetry. • The acid used predetermines completely different interaction with the AA2024 alloy.

  19. The effect of temperature on the toxicity of insecticides against Musca domestica L.: implications for the effective management of diarrhea.

    Directory of Open Access Journals (Sweden)

    Hafiz Azhar Ali Khan

    Full Text Available BACKGROUND: Diarrhea is an important cause of childhood mortality in developing countries like Pakistan because of unhygienic conditions, lack of awareness, and unwise use of preventive measures. Mechanical transmission of diarrheal pathogens by house flies, Musca domestica, is believed as the most effective route of diarrhea transmission. Although the use of insecticides as a preventive measure is common worldwide for the management of house flies, success of the measure could be compromised by the prevailing environmental temperature since it significantly affects toxicity of insecticides and thus their efficacy. Peaks of the house fly density and diarrheal cases are usually coincided and season specific, yet little is known about the season specific use of insecticides. METHODOLOGY/PRINCIPAL FINDINGS: To determine the temperature-toxicity relationship in house flies, the effect of post-bioassays temperature (range, 20-34°C on the toxicity of seven insecticides from organophosphate (chlorpyrifos, profenofos, pyrethroid (cypermethrin, deltamethrin and new chemical (emamectin benzoate, fipronil, spinosad classes was evaluated by using a feeding bioassay method. From 20-34°C, the toxicities of chlorpyrifos, profenofos, emamectin and fipronil increased 2.10, 2.93, 2.40 and 3.82 fold (i.e. positive temperature coefficient, respectively. Whereas, the toxicities of cypermethrin, deltamethrin and spinosad decreased 2.21, 2.42 and 3.16 fold (i.e. negative temperature coefficient, respectively. CONCLUSION/SIGNIFICANCE: These findings suggest that for the reduction in diarrheal cases, house flies should be controlled with insecticides according to the prevailing environmental temperature. Insecticides with a positive temperature coefficient may serve as potential candidates in controlling house flies and diarrhea epidemics in hot season and vice versa.

  20. Escalamiento para la producción de un biopesticida a partir de B. thuringiensis subsp. israelensis en biorreactores de 20 y 200 Litros

    Directory of Open Access Journals (Sweden)

    Sergio Ordúz

    2000-02-01

    Full Text Available

    Bacillus thuringiensis es una bacteria gram positiva esporulada que en fase estacionaria expresa algunas endotoxinas, contra un amplio espectro de larvas de insectos. Para desarrollar un biopesticida con base en B. thuringiensis que sea competitivo localmente es necesario generar la tecnología necesaria para su producción masiva, por esta razón el objetivo del trabajo fue escalar el proceso para la producción de B. thuringiensis basándose en el coeficiente volumétrico de transferencia de oxígeno corregido por presión (KLap partiendo de un biorreactor de 20 litros hasta llegar a un biorreactor de 200 litros.

    Para llevar a cabo este objetivo se realizó en primer lugar una evaluación del KLa a diferentes velocidades de agitación y flujos de aire por el método dinámico con el fin de obtener un modelo matemático que relacionara las variables anteriores. Se realizaron diferentes fermentaciones en un biorreactor de 20 litros con 11 litros de medio de cultivo utilizando B. thuringiensis subesp. israelensis. La velocidad de agitación y el flujo de aire se fijaron con base en un diseño factorial 22 con punto central realizando una repetición por cada punto del diseño, a continuación se realizaron dos fermentaciones adicionales por fuera del diseño para corroborar los resultados. Las fermentaciones de mayor escala se realizaron en un biorreactor de 200 litros con 110 litros de medio de cultivo, la velocidad de agitación y el flujo de aire se fijaron con base en los resultados obtenidos en las

  1. Insecticide Resistance Reducing Effectiveness of Malaria Control

    Centers for Disease Control (CDC) Podcasts

    2007-01-24

    Malaria prevention is increasingly insecticide based. Dr. John Gimnig, an entomologist with the Division of Parasitic Diseases, CDC, discusses evidence that mosquito resistance to insecticides, which is measured in the laboratory, could compromise malaria prevention in the field.  Created: 1/24/2007 by Emerging Infectious Diseases.   Date Released: 3/13/2007.

  2. Experimental immunologically mediated aplastic anemia (AA) in mice: cyclosporin A fails to protect against AA

    International Nuclear Information System (INIS)

    Knospe, W.H.; Steinberg, D.; Gratwohl, A.; Speck, B.

    1984-01-01

    Immunologically mediated aplastic anemia (AA) in mice was induced by the i.v. injection of 10(7) lymph node cells (LNC) from H-2k identical but Mls mismatched CBA/J donor mice into previously irradiated (600 rad total body gamma) C3H/HeJ mice. Cyclosporin A (CsA), 25 mg/kg, was administered subcutaneously from day -1 to day 30. Control mice included C3H/HeJ mice which received 600 rad alone, C3H/HeJ mice which received 600 rad plus CsA as above, and C3H/HeJ mice which received 600 rad total body irradiation followed by 10(7) LNC from CBA/J donors. CsA failed to prevent lethal AA. These results suggest that the pathogenetic mechanisms operating in immunologically mediated AA differ from the mechanisms operating in rodents transplanted with allogeneically mismatched marrow or spleen cells which develop graft-versus-host disease. The results are consistent with a non-T cell-dependent mechanism causing the AA

  3. Building materials and systems with vacuum insulation panels for external walls; Bauelemente und Systeme mit VIP fuer Aussenwandkonstruktionen - Schlussbericht

    Energy Technology Data Exchange (ETDEWEB)

    Binz, A; Steinke, G

    2007-07-01

    This final report for the Swiss Federal Office of Energy (SFOE) takes a look at materials and systems using vacuum insulation panels (VIP) for the construction of external walls. The aim of this research project was the development, practical use and market introduction of VIP systems that take account of the special properties of VIP. Along with partners in industry, applications involving external and internal insulation were examined. The need for protecting the vacuum panels against mechanical damage is stressed. The specific needs for the protection of external and internal applications are discussed. The dynamic developments in this relatively new area are commented on. Various mounting systems are examined and commented on. The thermal properties of such insulation systems and applications are noted and commented on.

  4. Differential roles of AVP and VIP signaling in the postnatal changes of neural networks for coherent circadian rhythms in the SCN

    Science.gov (United States)

    Ono, Daisuke; Honma, Sato; Honma, Ken-ichi

    2016-01-01

    The suprachiasmatic nucleus (SCN) is the site of the master circadian clock in mammals. The SCN neural network plays a critical role in expressing the tissue-level circadian rhythm. Previously, we demonstrated postnatal changes in the SCN network in mice, in which the clock gene products CRYPTOCHROMES (CRYs) are involved. Here, we show that vasoactive intestinal polypeptide (VIP) signaling is essential for the tissue-level circadian PER2::LUC rhythm in the neonatal SCN of CRY double-deficient mice (Cry1,2−/−). VIP and arginine vasopressin (AVP) signaling showed redundancy in expressing the tissue-level circadian rhythm in the SCN. AVP synthesis was significantly attenuated in the Cry1,2−/− SCN, which contributes to aperiodicity in the adult mice together with an attenuation of VIP signaling as a natural process of ontogeny. The SCN network consists of multiple clusters of cellular circadian rhythms that are differentially integrated by AVP and VIP signaling, depending on the postnatal period. PMID:27626074

  5. Characterization and selection of Bacillus thuringiensis isolates effective against Sitophilus oryzae Caracterização e seleção de isolados de Bacillus thuringiensis efetivos contra Sitophilus oryzae

    Directory of Open Access Journals (Sweden)

    Najara da Silva

    2010-08-01

    Full Text Available The entomopathogenic bacterium Bacillus thuringiensis is a control agent with toxic and environmental characteristics that allows the control of pest insects according to the Integrate Pest Management (IPM precepts. In order to find new strains, potentially toxic to Sitophilus oryzae L. 1763 (Coleoptera: Curculinidae, 1.073 strains of B. thuringiensis from parts of Brazil were used. Genetic material was extracted with InstaGene Matrix kit, used for the amplification of sequences in Polymerase chain reaction (PCR, and viewed in 1.5% agarose gel. The gene cry35Ba class was represented by 60 B. thuringiensis isolates (5.6%, which were then subjected to bioassays with S. oryzae larvae. Among the isolates studied, four caused more than 50% mortality in pathogenicity tests, and the isolates 544 and 622 were the most virulent, as determined by CL50 estimates. The four toxic isolates had spherical, bi-pyramidal and cuboid crystals, and a 44-kDa protein was found in sodium dodecyl sulphate - polyacrylamide gel electrophoresis (SDS-PAGE, which coded for the product of cry35Ba genes. These data demonstrate the potential of B. thuringiensis for the management of S. oryzae larvae.A bactéria entomopatogênica Bacillus thuringiensis (Bt é um agente de controle com características tóxicas e ambientais que permitem o controle de insetos-praga de acordo com as premissas do Manejo integrado de pragas (MIP. Com o objetivo de buscar novas linhagens potencialmente tóxicas para Sitophilus oryzae L. 1763 (Coleoptera: Curculinidae, caracterizaram-se molecularmente 1,073 isolados de B. thuringiensis de regiões do Brasil. O material genético foi extraído através do kit InstaGene Matrix, utilizado para a amplificação das seqüências através da técnica de Polymerase chain reaction PCR, sendo os resultados visualizados em gel de agarose 1,5%. A classe do gene cry35Ba foi representada por 60 isolados (5,6% de Bt, os quais foram submetidos a bioensaio com larvas

  6. Isolation and characterization of a novel native Bacillus thuringiensis strain BRC-HZM2 capable of degrading chlorpyrifos

    Czech Academy of Sciences Publication Activity Database

    Wu, S.; Peng, Y.; Huang, Z.; Huang, Z.; Xu, L.; Gelbič, Ivan; Guan, X.; Zhang, L.; Zou, S.

    2015-01-01

    Roč. 55, č. 3 (2015), s. 389-397 ISSN 0233-111X Institutional support: RVO:60077344 Keywords : Bacillus thuringiensis * biopesticide * biodegradation Subject RIV: EE - Microbiology, Virology Impact factor: 1.585, year: 2015 http://onlinelibrary.wiley.com/doi/10.1002/jobm.201300501/epdf

  7. Oil migration through unsaturated soils and its effect on the Vadose Zone Interactive Processes (VIP) model output

    International Nuclear Information System (INIS)

    Joseph, A.T.; Grenney, W.J.; Stevens, D.K.

    1994-01-01

    The VIP model, which simulates the concentration profiles of the hazardous compounds in the soil, water, and the air phases, assumes a fixed oily phase. The purpose of this study was to measure oil migration in soil systems and to determine its effect on the VIP model output. Experiments were conducted to demonstrate the mobility of an oil through the unsaturated zone of the soil. The studies were conducted in laboratory scale glass columns. A light petroleum oil and two types of soil were used. The experiments demonstrated that oil migrates down significantly through the soil columns. The extent of migration depended on the volume of oil applied and the type of soil. However, the applied oil was completely immobilized in the columns. The model was modified to incorporate oil migration. The modified model can be expected to produce more realistic contaminant concentration profiles during land treatment of oily wastes when compared to that produced by the present version of the VIP model. (Author)

  8. Larvicidal activities of chinaberry, neem and Bacillus thuringiensis israelensis (Bti to an insecticide resistant population of Anopheles arabiensis from Tolay, Southwest Ethiopia

    Directory of Open Access Journals (Sweden)

    Assalif Demissew

    2016-07-01

    Conclusions: Neem, chinaberry and Bti showed potent larvicidal and pupicidal activities. However, in the area, high level of mosquito resistance to pyrethroids and dichloro-diphenyl-tricgloroethane was seen which will pose serious challenge to vector control in the future. Therefore, using integrated approach including these botanical larvicides is warranted to manage insecticide resistance.

  9. Impact of Bollgard ® genetically modified cotton on the biodiversity ...

    African Journals Online (AJOL)

    Using cotton cultivars that express a gene of the Bacillus thuringiensis (Bt) bacterium producing a protein (Cry1Ac) with an insecticide effect on the Lepidoptera pests has made it possible to reduce the number of insecticide applications during the crop cycle. Thus, the objective was to determine, in the field during the ...

  10. Production of tannase by Aspergillus niger Aa-20 in submerged and solid-state fermentation: influence of glucose and tannic acid.

    Science.gov (United States)

    Aguilar, C N; Augur, C; Favela-Torres, E; Viniegra-González, G

    2001-05-01

    Tannase production by Aspergillus niger Aa-20 was studied in submerged (SmF) and solid-state (SSF) fermentation systems with different tannic acid and glucose concentrations. Tannase activity and productivity were at least 2.5 times higher in SSF than in SmF. Addition of high tannic acid concentrations increased total tannase activity in SSF, while in SmF it was decreased. In SmF, total tannase activity increased from 0.57 to 1.03 IU/mL, when the initial glucose concentration increased from 6.25 to 25 g/L, but a strong catabolite repression of tannase synthesis was observed in SmF when an initial glucose concentration of 50 g/L was used. In SSF, maximal values of total tannase activity decreased from 7.79 to 2.51 IU when the initial glucose concentration was increased from 6.25 to 200 g/L. Kinetic results on tannase production indicate that low tannase activity titers in SmF could be associated to an enzyme degradation process which is not present in SSF. Tannase titers produced by A. niger Aa-20 are fermentation system-dependent, favoring SSF over SmF.

  11. Resistance and behavioural response of Plutella xylostella (Lepidoptera: Plutellidae) populations to Bacillus thuringiensis formulations.

    Science.gov (United States)

    Zago, Hugo B; Siqueira, Herbert Á A; Pereira, Eliseu J G; Picanço, Marcelo C; Barros, Reginaldo

    2014-03-01

    Insecticide resistance is probably the major cause of control failure of Plutella xylostella (L.) in Brazil. In most production regions, the use of chemicals has been the prevalent method of control, with reduced efficacy through cropping seasons, even for the most recent use of products based on Bacillus thuringiensis (Bt). The current status of the resistance to these products was assessed, as well as the behavioural response of P. xylostella populations to Bt sprays. Most populations of P. xylostella were resistant to Bt products, particularly to Xentari®WDG (2-54-fold). Differences in walking characteristics of larvae were variable for most populations, for both Dipel®WP and Xentari®WDG, but not associated with resistance. Most females preferred to lay eggs on untreated surfaces and showed a reduced proportion of oviposition on treated surfaces that only correlated with resistance to Dipel®WP (r = -0.74, P = 0.02). Broad and indiscriminate use of Bt-based products has selected Brazilian P. xylostella populations to resistance. Larval movement appears to be a resistance-independent mechanism. Most populations of P. xylostella preferred to lay eggs on Bt-free surfaces, which might be a result of growers' practice of spraying the cabbage head. Reduced oviposition on treated surfaces correlated with physiological resistance, suggesting a behavioural response among the Bt-resistant colonies to Dipel®WP. © 2013 Society of Chemical Industry.

  12. Action of insecticidal N-alkylamides at site 2 of the voltage-sensitive sodium channel

    International Nuclear Information System (INIS)

    Ottea, J.A.; Payne, G.T.; Soderlund, D.M.

    1990-01-01

    Nine synthetic N-alkylamides were examined as inhibitors of the specific binding of [ 3 H]batrachotoxinin A 20α-benzoate ([ 3 H]BTX-B) to sodium channels and as activators of sodium uptake in mouse brain synaptoneurosomes. In the presence of scorpion (Leiurus quinquestriatus) venom, the six insecticidal analogues were active as both inhibitors of [ 3 H]BTX-B binding and stimulators of sodium uptake. These findings are consistent with an action of these compounds at the alkaloid activator recognition site (site 2) of the voltage-sensitive sodium channel. The three noninsecticidal N-alkylamides also inhibited [ 3 H]BTX-B binding but were ineffective as activators of sodium uptake. Concentration-response studies revealed that some of the insecticidal amides also enhanced sodium uptake through a second, high-affinity interaction that does not involve site 2, but this secondary effect does not appear to be correlated with insecticidal activity. The activities of N-alkylamides as sodium channel activators were influenced by the length of the alkenyl chain and the location of unsaturation within the molecule. These results further define the actions of N-alkylamides on sodium channels and illustrate the significance of the multiple binding domains of the sodium channel as target sites for insect control agents

  13. Mob/oriT, a mobilizable site-specific recombination system for unmarked genetic manipulation in Bacillus thuringiensis and Bacillus cereus.

    Science.gov (United States)

    Wang, Pengxia; Zhu, Yiguang; Zhang, Yuyang; Zhang, Chunyi; Xu, Jianyi; Deng, Yun; Peng, Donghai; Ruan, Lifang; Sun, Ming

    2016-06-10

    Bacillus thuringiensis and Bacillus cereus are two important species in B. cereus group. The intensive study of these strains at the molecular level and construction of genetically modified bacteria requires the development of efficient genetic tools. To insert genes into or delete genes from bacterial chromosomes, marker-less manipulation methods were employed. We present a novel genetic manipulation method for B. thuringiensis and B. cereus strains that does not leave selection markers. Our approach takes advantage of the relaxase Mob02281 encoded by plasmid pBMB0228 from Bacillus thuringiensis. In addition to its mobilization function, this Mob protein can mediate recombination between oriT sites. The Mob02281 mobilization module was associated with a spectinomycin-resistance gene to form a Mob-Spc cassette, which was flanked by the core 24-bp oriT sequences from pBMB0228. A strain in which the wild-type chromosome was replaced with the modified copy containing the Mob-Spc cassette at the target locus was obtained via homologous recombination. Thus, the spectinomycin-resistance gene can be used to screen for Mob-Spc cassette integration mutants. Recombination between the two oriT sequences mediated by Mob02281, encoded by the Mob-Spc cassette, resulted in the excision of the Mob-Spc cassette, producing the desired chromosomal alteration without introducing unwanted selection markers. We used this system to generate an in-frame deletion of a target gene in B. thuringiensis as well as a gene located in an operon of B. cereus. Moreover, we demonstrated that this system can be used to introduce a single gene or an expression cassette of interest in B. thuringiensis. The Mob/oriT recombination system provides an efficient method for unmarked genetic manipulation and for constructing genetically modified bacteria of B. thuringiensis and B. cereus. Our method extends the available genetic tools for B. thuringiensis and B. cereus strains.

  14. The colonization of Bacillus thuringiensis strains in bryophytes

    Czech Academy of Sciences Publication Activity Database

    Lin, Q.; Zhu, P.; Carballar-Lejarazú, R.; Gelbič, Ivan; Guan, X.; Xu, L.; Zhang, L.

    2017-01-01

    Roč. 41, č. 1 (2017), s. 41-48 ISSN 1300-0152 Institutional support: RVO:60077344 Keywords : Bacillus thuringiensis * GFP * plant colonization Subject RIV: EE - Microbiology, Virology OBOR OECD: Microbiology Impact factor: 1.038, year: 2016 http://journals.tubitak.gov.tr/biology/issues/biy-17-41-1/biy-41-1-5-1510-16.pdf

  15. The Synergist Effect of The Henna Plant, Lawsonia alba on Bacillus thuringiensis var. kurstaki Against Third Larval Instar of Pistachio Leaf Borer, Ocneria terebinthina Strg. (Lep.: Lymanteriidae

    Directory of Open Access Journals (Sweden)

    Zahra Sheibani

    2017-12-01

    Full Text Available Ocneria terebinthina Strg. (Lep.: Lymanteriidae is a leaf borer pest in pistachio orchard. The toxicity of Bacillus thuringiensis var. kurstaki alone and in combination with henna powder was investigated on third larval instar of O. terebinthina under laboratory conditions. Bioassay was carried out using spray technique on pistachio offshoot. Probit analysis of concentration-mortality data was conducted to estimate the LC50 value. The LC50 value of B. thuringiensis on third instar of larvae was estimated as 2817.30 ppm. The synergist effect of henna powder on efficacy of B. thuringiensis was also evaluated. Three concentrations of henna powder were combined with minimum lethal dose of B. thuringiensis against third larval instar. The results revealed that the henna powder had synergist effect on B. thuringiensis. The combination of B. thuringiensis and henna powder (6000 ppm caused 80.68% mortality, whereas B. thuringiensis alone caused 15.91% mortality. Our results suggested that henna powder can increase the efficacy of B. thuringiensis in controlling O. terebinthina in pest integrated management.

  16. Effects of Bacillus thuringiensis israelensis and spinosad on adult emergence of the non-biting midges Polypedilum nubifer (Skuse) and Tanytarsus curticornis Kieffer (Diptera: Chironomidae) in coastal wetlands.

    Science.gov (United States)

    Duchet, Claire; Franquet, Evelyne; Lagadic, Laurent; Lagneau, Christophe

    2015-05-01

    To optimize their efficacy, some insecticides used for mosquito control are introduced into aquatic ecosystems where mosquito larvae develop (marshes, ponds, sanitation devices) and cannot escape from the treated water. However, this raises the question of possible effects of mosquito larvicides on non-target aquatic species. Bacillus thuringiensis var. israelensis (Bti), which is well-known for its selectivity for Nematocera dipterans, is widely used for mosquito control all over the world. Spinosad, a mixture of spinosyns A and D known as fermentation products of a soil actinomycete (Saccharopolyspora spinosa), is a biological neurotoxic insecticide with a broader action spectrum. It is a candidate larvicide for mosquito control, but some studies showed that it may be toxic to beneficial or non-target species, including non-biting midges. The present study was therefore undertaken to assess the impact of Bti and spinosad on natural populations of Polypedilum nubifer (Skuse) and Tanytarsus curticornis Kieffer (Diptera: Chironomidae) in field enclosures implemented in Mediterranean coastal wetlands. Unlike Bti, spinosad had a strong lethal effect on P. nubifer and seems to affect T. curticornis at presumed recommended rates for field application. Differences in the sensitivity of these two species to spinosad confirm that population dynamics need to be known for a proper assessment of the risk encountered by chironomids in wetlands where larvicide-based mosquito control occurs. Copyright © 2015 Elsevier Inc. All rights reserved.

  17. Evaluation of toxicity of biorational insecticides against larvae of the alfalfa weevil

    Directory of Open Access Journals (Sweden)

    Gadi V.P. Reddy

    Full Text Available The alfalfa weevil, Hypera postica (Coleoptera: Curculionidae, is a major pest of alfalfa Medicago sativa L. (Fabaceae. While H. postica usually causes the most damage before the first cutting, in summer of 2015 damaging levels of the pest persisted in Montana well after the first harvest of alfalfa. Although conventional insecticides can control H. postica, these chemicals have adverse effects on non-target organisms including pollinators and natural enemy insects. In this context, use of biorational insecticides would be the best alternative options, as they are known to pose less risk to non-target organisms. We therefore examined the six commercially available biorational insecticides against H. postica under laboratory condition: Mycotrol® ESO (Beauveria bassiana GHA, Aza-Direct® (Azadirachtin, Met52® EC (Metarhizium brunneum F52, Xpectro OD® (B. bassiana GHA + pyrethrins, Xpulse OD® (B. bassiana GHA + Azadirachtin and Entrust WP® (spinosad 80%. Concentrations of 0.1, 0.5, 1.0, and 2.0 times the lowest labelled rates were tested for all products. However, in the case of Entrust WP, additional concentrations of 0.001 and 0.01 times the lowest label rate were also assessed. Mortality rates were determined at 1–9 days post treatment. Based on lethal concentrations and relative potencies, this study clearly showed that Entrust was the most effective, causing 100% mortality within 3 days after treatment among all the tested materials. With regard to other biorational, Xpectro was the second most effective insecticide followed by Xpulse, Aza-Direct, Met52, and Mycotrol. Our results strongly suggested that these biorational insecticides could potentially be applied for H. postica control. Keywords: Low risk insecticides, Insect pathogenic fungi, Efficacy, Lethal concentration, Mortality rate

  18. Potency of Bacillus thuringiensis isolates from bareng Tenes-Malang City as a biological control agent for suppressing third instar of Aedes aegypti larvae

    Science.gov (United States)

    Lutfiana, Nihayatul; Gama, Zulfaidah Penata

    2017-11-01

    Dengue is a mosquito-borne viral disease that is transmitted by the female Aedes species. The number of dengue fever cases has increased in many geographic regions including Indonesia and one of them occurred in Bareng Tenes, Malang City, East Java Province. The objective of this research was to identify the potency of B. thuringeinsis isolates from Bareng Tenes, Malang, as the biological agent to control third instar Ae. aegypti larvae and to identify the potential B. thuringiensis isolates based on 16S rDNA sequence. B. thuringiensis was isolated from water and soil from 12 sites in the Bareng Tenes area. Bacterial isolation was performed using B. thuringiensis selective media. Several isolates had similar phenotypic characters with B. thuringiensis used to toxicity test against third instar Ae. aegypti larvae. The LC50-96h value was determined using probit regression. The most effective isolate was identified based on the 16S rDNA sequence, then aligned to the reference isolate using the BLAST program. A phylogeny tree was constructed using the Maximum Likelihood method. This study showed that among 22 isolates of B. thuringiensis, only BA02b, BS04a, and BA03a isolates have similar phenotypic characters with B. thuringiensis. Based on the toxicity test of B. thuringiensis against the third instar of Ae. aegypti larvae, it was indicated that BA02b and BA03a isolates were the potential agents to control Ae. aegypti larvae. BA02b isolate was the most effective B. thuringiensis (LC50-96h = 2,75 x 107 cell/mL). Based on 16S rDNA sequence, BA02b was identified as Bacillus thuringiensis var. Israelensis BGSC4Q2 (99 % similarities).

  19. Insecticide resistance and glutathione S-transferases in mosquitoes ...

    African Journals Online (AJOL)

    Mosquito glutathione S-transferases (GSTs) have received considerable attention in the last 20 years because of their role in insecticide metabolism producing resistance. Many different compounds, including toxic xenobiotics and reactive products of intracellular processes such as lipid peroxidation, act as GST substrates.

  20. Visualization of the inferoposterior thoracic wall (VIP) and boomerang signs-novel sonographic signs of right pleural effusion.

    Science.gov (United States)

    Lau, James Siu Ki; Yuen, Chi Kit; Mok, Ka Leung; Yan, Wing Wa; Kan, Pui Gay

    2017-11-15

    This study is to present the diagnostic values of the novel sonographic visualization of the inferoposterior thoracic wall (VIP) and boomerang signs in detecting right pleural effusion by sonologists with little to no experience in ultrasound. A prospective analysis of a convenience sample of patients who were assessed by junior intensive care physicians was performed. The patients all underwent computed tomography (CT) of the chest or abdomen with lung bases as part of their care regardless of indication; the results were interpreted by radiologists and were considered the gold standard. Sonography was performed to assess for the presence of the VIP and boomerang signs. Sonographic and chest radiographic findings were compared against CT results. 73 patients were enrolled. The sensitivity and specificity for the VIP sign were 0.85 (95% confidence interval [CI], 0.67-0.94) and 0.86 (95% CI, 0.70-0.95). The sensitivity and specificity for the boomerang sign were 0.78 (95% CI, 0.60-0.90) and 0.87 (95% CI, 0.71-0.95). However, the sensitivity and specificity for the traditional approach of detecting an anechoic collection above the diaphragm to indicate pleural effusion were only 0.54 (95% CI, 0.37-0.71) and 0.86 (95% CI, 0.80-0.99). Despite inexperience in sonography, the novel VIP and boomerang signs show high diagnostic values in detecting right pleural effusion compared to the traditional methods. Copyright © 2017 Elsevier Inc. All rights reserved.