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Sample records for adult midgut microflora

  1. Midgut Malrotation Causing Intermittent Intestinal Obstruction in a Young Adult

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    Huseyin Kazim Bektasoglu; Ufuk Oguz Idiz; Mustafa Hasbahceci; Erkan Yardimci; Yurdakul Deniz Firat; Oguzhan Karatepe; Mahmut Muslumanoglu

    2014-01-01

    Midgut malrotation is a congenital anomaly of intestinal rotation and fixation that is generally seen in neonatal population. Adult cases are rarely reported. Early diagnosis is crucial to avoid life threatening complications. Here, we present an adulthood case of midgut volvulus as a rare cause of acute abdomen.

  2. Midgut malrotation causing intermittent intestinal obstruction in a young adult.

    Science.gov (United States)

    Bektasoglu, Huseyin Kazim; Idiz, Ufuk Oguz; Hasbahceci, Mustafa; Yardimci, Erkan; Firat, Yurdakul Deniz; Karatepe, Oguzhan; Muslumanoglu, Mahmut

    2014-01-01

    Midgut malrotation is a congenital anomaly of intestinal rotation and fixation that is generally seen in neonatal population. Adult cases are rarely reported. Early diagnosis is crucial to avoid life threatening complications. Here, we present an adulthood case of midgut volvulus as a rare cause of acute abdomen.

  3. Pyrosequencinq analysis of the oral microflora of healthy adults

    NARCIS (Netherlands)

    Keijser, B.J.F.; Zaura, E.; Huse, S.M.; Vossen, J.M.B.M. van der; Schuren, F.H.J.; Montijn, R.C.; Gate, J.M. ten; Crielaard, W.

    2008-01-01

    A good definition of commensal microflora and an understanding of its relation to health are essential in preventing and combating disease. We hypothesized that the species richness of human oral microflora is underestimated. Saliva and supragingival plaque were sampled from 71 and 98 healthy adults

  4. Intestinal stem cells in the adult Drosophila midgut

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    Jiang, Huaqi, E-mail: Huaqi.Jiang@UTSouthwestern.edu [Department of Developmental Biology, UT Southwestern Medical Center, 6000 Harry Hines Blvd., Dallas, TX, 75235 (United States); Edgar, Bruce A., E-mail: b.edgar@dkfz.de [ZMBH-DKFZ Alliance, Im Neuenheimer Feld 282, D-69120 Heidelberg (Germany); Division of Basic Sciences, Fred Hutchinson Cancer Research Center, 1100 Fairview Ave. N., Seattle, WA 98109 (United States)

    2011-11-15

    Drosophila has long been an excellent model organism for studying stem cell biology. Notably, studies of Drosophila's germline stem cells have been instrumental in developing the stem cell niche concept. The recent discovery of somatic stem cells in adult Drosophila, particularly the intestinal stem cells (ISCs) of the midgut, has established Drosophila as an exciting model to study stem cell-mediated adult tissue homeostasis and regeneration. Here, we review the major signaling pathways that regulate the self-renewal, proliferation and differentiation of Drosophila ISCs, discussing how this regulation maintains midgut homeostasis and mediates regeneration of the intestinal epithelium after injury. -- Highlights: Black-Right-Pointing-Pointer The homeostasis and regeneration of adult fly midguts are mediated by ISCs. Black-Right-Pointing-Pointer Damaged enterocytes induce the proliferation of intestinal stem cells (ISC). Black-Right-Pointing-Pointer EGFR and Jak/Stat signalings mediate compensatory ISC proliferation. Black-Right-Pointing-Pointer Notch signaling regulates ISC self-renewal and differentiation.

  5. Primary adult midgut volvulus mimicking acute appendicitis: A case report and review of the literature

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    Jeffrey J. Leow

    2016-01-01

    Conclusion: While rare, adult midgut volvulus has the potential for morbidity and mortality if not diagnosed early with intestinal ischaemia and related sequlae. Clinicians should consider this and if in doubt, perform an early contrast CT scan.

  6. Diversity of the bacterial and fungal microflora from the midgut and cuticle of phlebotomine sand flies collected in North-Western Iran.

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    Mohammad Akhoundi

    Full Text Available BACKGROUND: Phlebotomine sand flies are the vectors of the leishmaniases, parasitic diseases caused by Leishmania spp. Little is known about the prevalence and diversity of sand fly microflora colonizing the midgut or the cuticle. Particularly, there is little information on the fungal diversity. This information is important for development of vector control strategies. METHODOLOGY/PRINCIPAL FINDINGS: FIVE SAND FLY SPECIES: Phlebotomus papatasi, P. sergenti, P. kandelakii, P. perfiliewi and P. halepensis were caught in Bileh Savar and Kaleybar in North-Western Iran that are located in endemic foci of visceral leishmaniasis. A total of 35 specimens were processed. Bacterial and fungal strains were identified by routine microbiological methods. We characterized 39 fungal isolates from the cuticle and/or the midgut. They belong to six different genera including Penicillium (17 isolates, Aspergillus (14, Acremonium (5, Fusarium (1, Geotrichum (1 and Candida (1. We identified 33 Gram-negative bacteria: Serratia marcescens (9 isolates, Enterobacter cloacae (6, Pseudomonas fluorescens (6, Klebsiella ozaenae (4, Acinetobacter sp. (3, Escherichia coli (3, Asaia sp. (1 and Pantoea sp. (1 as well as Gram-positive bacteria Bacillus subtilis (5 and Micrococcus luteus (5 in 10 isolates. CONCLUSION/SIGNIFICANCE: Our study provides new data on the microbiotic diversity of field-collected sand flies and for the first time, evidence of the presence of Asaia sp. in sand flies. We have also found a link between physiological stages (unfed, fresh fed, semi gravid and gravid of sand flies and number of bacteria that they carry. Interestingly Pantoea sp. and Klebsiella ozaenae have been isolated in Old World sand fly species. The presence of latter species on sand fly cuticle and in the female midgut suggests a role for this arthropod in dissemination of these pathogenic bacteria in endemic areas. Further experiments are required to clearly delineate the vectorial

  7. Preoperative sonographic diagnosis of midgut malrotation with volvulus in adults: the "whirlpool" sign.

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    Yeh, W C; Wang, H P; Chen, C; Wang, H H; Wu, M S; Lin, J T

    1999-06-01

    Midgut malrotation and volvulus, found mostly in children, are rare and difficult to diagnose preoperatively in adults. We report 2 cases in which a 68-year-old man and a 75-year-old woman presented with intermittent cramping abdominal pain, abdominal distention, and vomiting. Abdominal sonography demonstrated wrapping of the superior mesenteric vein and bowel loops around the superior mesenteric artery (the "whirlpool sign") in both patients. Abdominal CT revealed similar findings. The diagnoses of midgut volvulus and mesenteric malrotation were made, and the patients underwent laparotomy. The man was confirmed to have duodenojejunal malrotation and volvulus, and the woman had cecal volvulus. The whirlpool sign is valuable for the preoperative diagnosis of mesenteric vessel malrotation and midgut volvulus.

  8. Barber Pole Sign in CT Angiography, Adult Presentation of Midgut Malrotation: A Case Report

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    Garcelan-Trigo

    2015-07-01

    Full Text Available Adult midgut volvulus is a challenging diagnosis because of its low incidence and nonspecific symptoms. Diagnostic delay and long-term complaints are frequent in this clinical scenario. We reported a patient referred to our diagnostic imaging unit with intermittent abdominal pain, bloating and episodic vomiting for several years. He underwent barium gastrointestinal transit and abdominal ultrasound, which revealed severe gastric dilatation, food retention and slow transit until a depressed duodenojejunal flexure, with malrotation of the midgut and jejunal loops being located in the right upper quadrant. Computed tomography angiography was performed, showing rotation of the small intestine around the mesentery root, suggestive of midgut malrotation. In addition, an abnormal twisted disposition of superior mesenteric artery with corkscrew appearance was seen, shaping the pole-barber sign which was evident in volume rendering three-dimensional reconstructions. The patient underwent scheduled surgical treatment without any complication and had good outcome after hospital discharge and follow-up. Computed tomography plays an important role in evaluation of adult midgut volvulus. In addition, angiographic reconstructions can help us to assess the anatomic disposition of mesenteric vascular supply. Both of these assessments are useful in preoperative management.

  9. Barber Pole Sign in CT Angiography, Adult Presentation of Midgut Malrotation: A Case Report

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    Garcelan-Trigo, Juan Arsenio; Tello-Moreno, Manuel; Rabaza-Espigares, Manuel Jesus; Talavera-Martinez, Ildefonso

    2015-01-01

    Adult midgut volvulus is a challenging diagnosis because of its low incidence and nonspecific symptoms. Diagnostic delay and long-term complaints are frequent in this clinical scenario. We reported a patient referred to our diagnostic imaging unit with intermittent abdominal pain, bloating and episodic vomiting for several years. He underwent barium gastrointestinal transit and abdominal ultrasound, which revealed severe gastric dilatation, food retention and slow transit until a depressed duodenojejunal flexure, with malrotation of the midgut and jejunal loops being located in the right upper quadrant. Computed tomography angiography was performed, showing rotation of the small intestine around the mesentery root, suggestive of midgut malrotation. In addition, an abnormal twisted disposition of superior mesenteric artery with corkscrew appearance was seen, shaping the pole-barber sign which was evident in volume rendering three-dimensional reconstructions. The patient underwent scheduled surgical treatment without any complication and had good outcome after hospital discharge and follow-up. Computed tomography plays an important role in evaluation of adult midgut volvulus. In addition, angiographic reconstructions can help us to assess the anatomic disposition of mesenteric vascular supply. Both of these assessments are useful in preoperative management. PMID:26557278

  10. EPIGLOTTIS MICROFLORA OF ADULT PATIENTS WITH ACUTE EPIGLOTTITIS

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    Golovko NA

    2016-12-01

    Full Text Available Introduction. Nowadays acute infectious-inflammatory processes of upper respiratory tract, including acute epiglottitis retain a high proportion among human pathology. In the literature acute epiglottitis is allocated into an independent nosology as severe acute phlegmonous bacterial inflammation of the epiglottis and hypopharynx. There are currently no clear guidelines on how to classify an acute epiglottitis, as well as protocols for patients at various stages of the pathological process. According to common belief, Haemophilus influenzae type -B (Haemophilus influenza type b (Hib is the most common cause of epiglottitis. At present, the main etiological role in the genesis of acute epiglottitis in children belongs to haemophilus influenzae. In adults the causes of the disease are beta hemolytic streptococci groups A, B, pneumococcus, Klebsiella, Pseudomonas, Staphylococcus aureus, herpes simplex virus (type 1 and parainfluenza, and others.The aim of this work is to study: the mucosal microflora of the epiglottis in adult patients with acute epiglottitis and to study sensitivity of certain isolates to antimicrobial agents. Material & methods. 86 adult patients with acute epiglottitis were observed: 36 with abscess form of epiglottitis and 50 - with infiltrative. Microbiological analysis of mucosal swab samples taken from hypopharynx were conducted by the conventional technology: for seeding solid or liquid nutrient medium, followed by allocation of isolith and its microscopic, biochemical and serological identification. Microorganisms were classified according to schemes of Bergy. Antimicrobial susceptibility of each strain was determined in accordance with the guidelines. We used discs with antibacterial drugs. The availability of sensitive and resistant strains of microorganisms to antibiotics was assessed. A mucous membrane of the epiglottis was analyzed through microbiological investigation in 86 patients with acute epiglottitis. As a

  11. Conserved mechanisms of tumorigenesis in the Drosophila adult midgut.

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    Òscar Martorell

    Full Text Available Whereas the series of genetic events leading to colorectal cancer (CRC have been well established, the precise functions that these alterations play in tumor progression and how they disrupt intestinal homeostasis remain poorly characterized. Activation of the Wnt/Wg signaling pathway by a mutation in the gene APC is the most common trigger for CRC, inducing benign lesions that progress to carcinomas due to the accumulation of other genetic alterations. Among those, Ras mutations drive tumour progression in CRC, as well as in most epithelial cancers. As mammalian and Drosophila's intestines share many similarities, we decided to explore the alterations induced in the Drosophila midgut by the combined activation of the Wnt signaling pathway with gain of function of Ras signaling in the intestinal stem cells. Here we show that compound Apc-Ras clones, but not clones bearing the individual mutations, expand as aggressive intestinal tumor-like outgrowths. These lesions reproduce many of the human CRC hallmarks such as increased proliferation, blockade of cell differentiation and cell polarity and disrupted organ architecture. This process is followed by expression of tumoral markers present in human lesions. Finally, a metabolic behavioral assay shows that these flies suffer a progressive deterioration in intestinal homeostasis, providing a simple readout that could be used in screens for tumor modifiers or therapeutic compounds. Taken together, our results illustrate the conservation of the mechanisms of CRC tumorigenesis in Drosophila, providing an excellent model system to unravel the events that, upon mutation in Apc and Ras, lead to CRC initiation and progression.

  12. Requirement of matrix metalloproteinase-1 for intestinal homeostasis in the adult Drosophila midgut

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    Lee, Shin-Hae; Park, Joung-Sun [Department of Molecular Biology, College of Natural Science, Pusan National University, Busan 609-735 (Korea, Republic of); Kim, Young-Shin [Research Institute of Genetic Engineering, Pusan National University, Busan 609-735 (Korea, Republic of); Chung, Hae-Young [Molecular Inflammation Research Center for Aging Intervention (MRCA), College of Pharmacy, Pusan National University, Busan 609-735 (Korea, Republic of); Yoo, Mi-Ae, E-mail: mayoo@pusan.ac.kr [Department of Molecular Biology, College of Natural Science, Pusan National University, Busan 609-735 (Korea, Republic of)

    2012-03-10

    Stem cells are tightly regulated by both intrinsic and extrinsic signals as well as the extracellular matrix (ECM) for tissue homeostasis and regenerative capacity. Matrix metalloproteinases (MMPs), proteolytic enzymes, modulate the turnover of numerous substrates, including cytokine precursors, growth factors, and ECM molecules. However, the roles of MMPs in the regulation of adult stem cells are poorly understood. In the present study, we utilize the Drosophila midgut, which is an excellent model system for studying stem cell biology, to show that Mmp1 is involved in the regulation of intestinal stem cells (ISCs). The results showed that Mmp1 is expressed in the adult midgut and that its expression increases with age and with exposure to oxidative stress. Mmp1 knockdown or Timp-overexpressing flies and flies heterozygous for a viable, hypomorphic Mmp1 allele increased ISC proliferation in the gut, as shown by staining with an anti-phospho-histone H3 antibody and BrdU incorporation assays. Reduced Mmp1 levels induced intestinal hyperplasia, and the Mmp1depletion-induced ISC proliferation was rescued by the suppression of the EGFR signaling pathway, suggesting that Mmp1 regulates ISC proliferation through the EGFR signaling pathway. Furthermore, adult gut-specific knockdown and whole-animal heterozygotes of Mmp1 increased additively sensitivity to paraquat-induced oxidative stress and shortened lifespan. Our data suggest that Drosophila Mmp1 is involved in the regulation of ISC proliferation for maintenance of gut homeostasis. -- Highlights: Black-Right-Pointing-Pointer Mmp1 is expressed in the adult midgut. Black-Right-Pointing-Pointer Mmp1 is involved in the regulation of ISC proliferation activity. Black-Right-Pointing-Pointer Mmp1-related ISC proliferation is associated with EGFR signaling. Black-Right-Pointing-Pointer Mmp1 in the gut is required for the intestinal homeostasis and longevity.

  13. Study on Fungal Flora in the Midgut of the Larva and Adult of the Different Populations of the Malaria Vector Anopheles stephensi

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    L Tajedin

    2009-07-01

    Full Text Available Abstract Background: Many microorganisms in midgut of mosquito challenge with their host and also other pathogens pre­sent in midgut. The aim of this study was presence of non-pathogens microorganisms like fungal flora which may be cru­cial on interaction between vectors and pathogens."nMethods: Different populations of Anopheles stephensi were reared in insectary and objected to determine fungal flora in their midguts. The midgut paunch of mosquito adults and larvae as well as breading water and larval food sam­ples transferred on Subaru-dextrose agar, in order to detect the environment fungus."nResults: Although four fungi, Aspergillus, Rhizopus, Geotrichum and Sacharomyces were found in the food and wa­ter, but only Aspiragilus observed in the midgut of larvae. No fungus was found in the midgut of adults. This is the first report on fungal flora in the midgut of the adults and larvae of An. stephensi and possible stadial transmission of fungi from immature stages to adults."nConclusion: The midgut environment of adults is not compatible for survivorship of fungi but the larval midgut may con­tain few fungi as a host or even pathogen.

  14. Debra-mediated Ci degradation controls tissue homeostasis in Drosophila adult midgut.

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    Li, Zhouhua; Guo, Yueqin; Han, Lili; Zhang, Yan; Shi, Lai; Huang, Xudong; Lin, Xinhua

    2014-02-11

    Adult tissue homeostasis is maintained by resident stem cells and their progeny. However, the underlying mechanisms that control tissue homeostasis are not fully understood. Here, we demonstrate that Debra-mediated Ci degradation is important for intestinal stem cell (ISC) proliferation in Drosophila adult midgut. Debra inhibition leads to increased ISC activity and tissue homeostasis loss, phenocopying defects observed in aging flies. These defects can be suppressed by depleting Ci, suggesting that increased Hedgehog (Hh) signaling contributes to ISC proliferation and tissue homeostasis loss. Consistently, Hh signaling activation causes the same defects, whereas depletion of Hh signaling suppresses these defects. Furthermore, the Hh ligand from multiple sources is involved in ISC proliferation and tissue homeostasis. Finally, we show that the JNK pathway acts downstream of Hh signaling to regulate ISC proliferation. Together, our results provide insights into the mechanisms of stem cell proliferation and tissue homeostasis control.

  15. Midgut volvulus: a rare cause of episodes of intestinal obstruction in an adult; Volvulo de intestino medio: una rara causa de crisis oclusivas en el adulto

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    Palomo, V.; Higuera, A.; Munoz, R.; Sanchez, F. [Hospital Alto Guadalquivir. Andujar. Jaen (Spain)

    2002-07-01

    Midgut volvulus occurs frequently in infants and children, but is uncommon in adults. We present a case of intestinal malrotation complicated by midgut volvulus in a young woman who complained of chronic intermittent abdominal pain of increasing intensity. The radiologies diagnosis was based mainly on upper gastrointestinal barium study, and was confirmed intraoperatively. (Author) 11 refs.

  16. Sexual differences in destructive capability and midgut enzyme activities in adult variegated grasshoppers Zonocerus variegatus (LINNAEUS, 1758 (Orthoptera: Pyrgomorphidae

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    Ademolu Kehinde O.

    2016-12-01

    Full Text Available The variegated grasshopper Zonocerus variegatus is a polyphagous insect, feeding on numerous food and cash crops. The present study aimed to investigate the sexual variations in the destructive capability of the adult insects and the composition of leaves damaged by them, as well as in the levels of midgut microbial flora and digestive enzymes (cellulase, amylase and α-glucosidase. The results showed that females consumed and caused more damage to cassava leaves than their male congeners. The leaves damaged by males contained more nutrients than those damaged by females. The gut microbial flora and enzyme assay showed that females had significantly larger colony forming units and a non-significant difference in enzyme activities. It can thus be concluded that adult females are more destructive than males.

  17. Biochemical characterization of pectinase activity from the digestive midgut fluid of larvae and adult of the Colorado potato beetle, Leptinotarsa decemlineata (Col: Chrysomelidae

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    Razieh Karimi

    2015-03-01

    Full Text Available The study of pectinase enzyme in potato leaf beetle that is the most important pest of potatoes consider as an effective way to develop control methods because this enzyme is important in degrading plant cell wall. Pectinase enzyme was studied in midgut of Colorado potato beetle (CPB. This enzyme was extracted from the midgut of larvae and adult of CPB and then their important features were examined by specific substrate, pectin 1 percent. The optimum enzyme activity in 4th instar larvae and adult occurred at pH 5-6 range. Effect of temperature on enzyme activity were examined, the results suggest that the pectinase in midgut of 4th instar larvae and adult shows maximum activity at temperature 40 Celsius degree and 35 Celsius degree, respectively. Zymogram analysis showed presence of two activity bands for pectinase enzyme. The effect of various chemical compounds on the activity of enzyme showed that SDS, Urea and Tris reduced the enzyme activity. NaCl and CaCl2 increased this enzyme activity in 4th instar larvae and adult of CPB. This is first report of pectinase activity in L. decemlineata.

  18. A subset of neurons controls the permeability of the peritrophic matrix and midgut structure in Drosophila adults.

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    Kenmoku, Hiroyuki; Ishikawa, Hiroki; Ote, Manabu; Kuraishi, Takayuki; Kurata, Shoichiro

    2016-08-01

    The metazoan gut performs multiple physiological functions, including digestion and absorption of nutrients, and also serves as a physical and chemical barrier against ingested pathogens and abrasive particles. Maintenance of these functions and structures is partly controlled by the nervous system, yet the precise roles and mechanisms of the neural control of gut integrity remain to be clarified in Drosophila Here, we screened for GAL4 enhancer-trap strains and labeled a specific subsets of neurons, using Kir2.1 to inhibit their activity. We identified an NP3253 line that is susceptible to oral infection by Gram-negative bacteria. The subset of neurons driven by the NP3253 line includes some of the enteric neurons innervating the anterior midgut, and these flies have a disorganized proventricular structure with high permeability of the peritrophic matrix and epithelial barrier. The findings of the present study indicate that neural control is crucial for maintaining the barrier function of the gut, and provide a route for genetic dissection of the complex brain-gut axis in adults of the model organism Drosophila.

  19. Morphology of the midgut of Rhipicephalus sanguineus (Latreille, 1806) (Acari: Ixodidae) adult ticks in different feeding stages.

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    Remedio, R N; Sampieri, B R; Vendramini, M C R; Souza, N M; Anholeto, L A; Denardo, T A G B; Camargo-Mathias, M I

    2013-01-01

    The intestinal epithelial cells of ticks are fundamental for their full feeding and reproductive success, besides being considered important sites for the development of pathogens. Rhipicephalus sanguineus ticks are known for their great medical and veterinary importance, and for this reason, the knowledge of their intestinal morphology may provide relevant subsidies for the control of these animals, either by direct acaricidal action over these cells or by the production of vaccines. Therefore, this study aimed to describe the midgut morphology of male and female R. sanguineus ticks in different feeding stages, by means of histological analysis. Significant differences were observed between the genders, and such alterations may refer mainly to the distinct demands for nutrients, much higher in females, which need to develop and carry out the egg-laying process. In general, the midgut is coated by a thin muscle layer and presents a pseudostratified epithelium, in which two basic types of cells can be observed, connected to a basal membrane-generative or stem and digestive cells. The latter was classified as follows: residual, deriving from the phase anterior to ecdysis; pinocytic, with vesicles containing liquid or pre-digested components of blood; phagocytic, with entire cells or remnants of nuclear material inside cytoplasmic vesicles; and mature, free in the lumen. Digestion is presumably intracellular and asynchronous and corresponds to a process which starts with the differentiation of generative cells into pinocytic digestive cells, which subsequently start to phagocytize intact blood cells and finally detach from the epithelium, being eliminated with feces.

  20. Midgut malrotation with chronic abdominal pain

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    Anil K Wanjari

    2012-01-01

    Full Text Available Abnormalities in midgut rotation occur during the physiological herniation of midgut between the 5 th and 10 th week of gestation. The most significant abnormality is narrow small bowel mesentery which is prone to volvulus. This occurs most frequently in the neonatal period, less commonly midgut malrotation presents in adulthood with either acute volvulus or chronic abdominal symptoms. It is the latter group that represents a diagnostic challenge. We report a case of a 17-year-old male patient who presented with 10-year history of nonspecific gastro-intestinal symptoms. After extensive investigation the patient was diagnosed with midgut malrotation following computed tomography of abdomen. The patient was treated with a laparoscopic Ladd′s procedure and at 3 months he was gaining weight and had stopped vomiting. A laparoscopic Ladd′s procedure is an acceptable alternative to the open technique in treating symptomatic malrotation in adults. Midgut malrotation is a rare congenital anomaly which may present as chronic abdominal pain. Abdominal CT is helpful for diagnosis.

  1. Protein requirements in larvae and adults of Scaptotrigona postica (Hymenoptera: Apidae [correction of Apidia], Meliponinae): midgut proteolytic activity and pollen digestion.

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    do Carmo Zerbo, A; Silva de Moraes, R L; Brochetto-Braga, M R

    2001-05-01

    The number and degree of digestion of pollen grains in the midgut and rectum, the midgut proteolytic activity and the time of pollen grain passage through the digestive tract in the stingless bee Scaptotrigona postica (Latreille) have been analyzed. The results show similar protein requirements among larvae, nurse bees and queens, as well as between forager bees and old males, but these requirements are higher in individuals from the former groups than in those from the latter. Although protein requirements have been demonstrated to vary according to a bee's activity in the colony, they are similar among bees from different castes or sexes. These changes in feeding behavior are related to the bee's function and to less competition for nourishment among individuals of the colony. It is also noted that pollen grains took between 6 and 28 h to pass through the digestive tract. Pollen grains are irregularly accumulated in the various regions of the midgut, which may reflect functional differentiation throughout the midgut.

  2. Midgut morphological changes and autophagy during metamorphosis in sand flies.

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    Malta, Juliana; Heerman, Matthew; Weng, Ju Lin; Fernandes, Kenner M; Martins, Gustavo Ferreira; Ramalho-Ortigão, Marcelo

    2017-03-11

    During metamorphosis, holometabolous insects undergo significant remodeling of their midgut and become able to cope with changes in dietary requirements between larval and adult stages. At this stage, insects must be able to manage and recycle available food resources in order to develop fully into adults, especially when no nutrients are acquired from the environment. Autophagy has been previously suggested to play a crucial role during metamorphosis of the mosquito. Here, we investigate the overall morphological changes of the midgut of the sand fly during metamorphosis and assess the expression profiles of the autophagy-related genes ATG1, ATG6, and ATG8, which are associated with various steps of the autophagic process. Morphological changes in the midgut start during the fourth larval instar, with epithelial degeneration followed by remodeling via the differentiation of regenerative cells in pre-pupal and pupal stages. The changes in the midgut epithelium are paired with the up-regulation of ATG1, ATG6 and ATG8 during the larva-adult transition. Vein, a putative epidermal growth factor involved in regulating epithelial midgut regeneration, is also up-regulated. Autophagy has further been confirmed in sand flies via the presence of autophagosomes residing within the cytoplasmic compartment of the pupal stages. An understanding of the underlying mechanisms of this process should aid the future management of this neglected tropical vector.

  3. Transcriptional profiling of midgut immunity response and degeneration in the wandering silkworm, Bombyx mori.

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    Qiuyun Xu

    Full Text Available BACKGROUND: Lepidoptera insects have a novel development process comprising several metamorphic stages during their life cycle compared with vertebrate animals. Unlike most Lepidoptera insects that live on nectar during the adult stage, the Bombyx mori silkworm adults do not eat anything and die after egg-laying. In addition, the midguts of Lepidoptera insects produce antimicrobial proteins during the wandering stage when the larval tissues undergo numerous changes. The exact mechanisms responsible for these phenomena remain unclear. PRINCIPAL FINDINGS: We used the silkworm as a model and performed genome-wide transcriptional profiling of the midgut between the feeding stage and the wandering stage. Many genes concerned with metabolism, digestion, and ion and small molecule transportation were down-regulated during the wandering stage, indicating that the wandering stage midgut loses its normal functions. Microarray profiling, qRT-PCR and western blot proved the production of antimicrobial proteins (peptides in the midgut during the wandering stage. Different genes of the immune deficiency (Imd pathway were up-regulated during the wandering stage. However, some key genes belonging to the Toll pathway showed no change in their transcription levels. Unlike butterfly (Pachliopta aristolochiae, the midgut of silkworm moth has a layer of cells, indicating that the development of midgut since the wandering stage is not usual. Cell division in the midgut was observed only for a short time during the wandering stage. However, there was extensive cell apoptosis before pupation. The imbalance of cell division and apoptosis probably drives the continuous degeneration of the midgut in the silkworm since the wandering stage. CONCLUSIONS: This study provided an insight into the mechanism of the degeneration of the silkworm midgut and the production of innate immunity-related proteins during the wandering stage. The imbalance of cell division and apoptosis

  4. METABOLIC EFFECT OF FOS (FRUCTOOLIGOSACCHARIDE IN TERMS OF GUT INCRETIN (GLP-1 GUT MICROFLORA AND WEIGHT REDUCTION IN OBESE ADULTS

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    Mini K.Sheth

    2014-09-01

    Full Text Available In the recent years, obesity has increased beyond imagination. Appropriate dietary strategies which have the potential for weight loss demand patience and strong determination on part of the individual, however inclusion of functional foods like FOS that modulate gut hormones have a promising role in weight management. Methods: A randomized double-blind placebo-controlled trial was used as the study design wherein 65 obese adults were divided into experimental group (which was given 12 g of FOS and a placebo group (which was fed with 12 g dextrose. The subjects were given the supplements daily for 12 week period. Their plasma samples were anlaysed for GLP-1 and microbial count in fecal samples were determined in terms of lactic acid bacteria, bifidobacteria and enteric pathogens. Hunger scores, dietary intake, and anthropometric parameters were assessed using standard techniques. Results: FOS supplementation resulted in improved plasma GLP-1 level by 17.0%. Significant improvement was observed in hunger score by 3.15% (p<0.05 along with reduction in dietary intake of energy (kcal by 8%, carbohydrate (g by 8%, protein (g by 6% and fat (g by 2%. Further, reductions were observed in total body weight (kg, BMI, % body fat and waist circumference (cm levels by 4%, 1.06%, 4% and 1.66% respectively (p<0.001, p<0.001, p<0.001, p<0.05. The mean log counts of beneficial gut microbiota i.e. lactic acid bacteria and bifidobacteria increased significantly by 14 % and 10 % respectively along with 20% reduction in enteric pathogen. Conclusion: Daily intake of 12 gm FOS for 12 weeks helps in improving gut health and weight loss through increased satiety in obese individuals.

  5. Perforated midgut diverticulitis: Revisited

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    Milan Spasojevic; Jens Marius Naesgaard; Dejan Ignjatovic

    2012-01-01

    AIM:To study and provide data on the evolution of medical procedures and outcomes of patients suffering from perforated midgut diverticulitis.METHODS:Three data sources were used:the Medline and Google search engines were searched for case reports on one or more patients treated for perforated midgut diverticulitis (Meckel's diverticulitis excluded)that were published after 1995.The inclusion criterion was sufficient individual patient data in the article.Both indexed and non-indexed journals were used.Patients treated for perforated midgut diverticulitis at Vestfold Hospital were included in this group.Data on symptoms,laboratory and radiology results,treatment modalities,surgical access,procedures,complications and outcomes were collected.The Norwegian patient registry was searched to find patients operated upon for midgut diverticulitis from 1999 to 2007.The data collected were age,sex,mode of access,surgical procedure performed and number of patients per year.Historical controls were retrieved from an article published in 1995 containing pertinent individual patient data.Statistical analysis was done with SPSS software.RESULTS:Group I:106 patients (48 men) were found.Mean age was 72.2 ± 13.1 years (mean ± SD).Age or sex had no impact on outcomes (P =0.057 and P =0.771,respectively).Preoperative assessment was plain radiography in 53.3% or computed tomography (CT)in 76.1%.Correct diagnosis was made in 77.1% with CT,5.6% without (P =0.001).Duration of symptoms before hospitalization was 3.6 d (range:1-35 d),but longer duration was not associated with poor outcome (P =0.748).Eighty-six point eight percent of patients underwent surgery,92.4% of these through open access where 90.1% had bowel resection.Complications occurred in 19.2% of patients and 16.3% underwent reoperation.Distance from perforation to Treitz ligament was 41.7 ± 28.1 cm.At surgery,no peritonitis was found in 29.7% of patients,local peritonitis in 47.5%,and diffuse

  6. [MICROFLORA AND ORAL DISEASE].

    Science.gov (United States)

    Khavkin, A I; Ippolitov, Y A; Aleshina, E O; Komarova O N

    2015-01-01

    Acid-producing microorganisms are base etiological agents of lesions of tooth enamel and destruction of dentin. The process start by specific microflora of tooth deposit--Streptococcus mutans, Lactobacteria and Actinomycetis viscosus which ferment food carbogydrate to form acids. High titre of them in oral cavity may be considered like a marker of carbohydrate food. But the pathogenic bacteria don't have aggression to host organism until they will have virulent factors which help to get over protection of host organism. At the same time, microflora of oral cavity is involved to form pellicula. Pellicula is a biofilm which to protect tooth enamel and dentin. Understanding relationships between safety factors of host and pathogenic microflora of oral cavity will give to create effective methods of prevention and treatment.

  7. Effects of New Dietary Fiber from Japanese Apricot (Prunus mume Sieb. et Zucc. on Gut Function and Intestinal Microflora in Adult Mice

    Directory of Open Access Journals (Sweden)

    Nobuki Gato

    2011-03-01

    Full Text Available Much attention has been focused recently on functional foods. Ume, the Japanese name for the apricot of Prunus mume Sieb. et Zucc., is an example of a Japanese traditional functional food. There are, however, few reports on the effects of fiber from this fruit on bowel function. With this objective, we prepared ume fiber to test the hypothesis that it can change gut function and intestinal flora in mice. Mice were fed an ume fiber (UF or cellulose (CF diet (control for 40 days. The fecal weight, fecal lipids, plasma lipids and cecal composition of the microflora were analyzed. The amount of feces was significantly greater in the UF group than in the CF group (p < 0.01. The fecal lipids content (% DW of the feces sampled on the final days of the experiment were significantly greater in the UF group than in the CF group (p < 0.01. Plasma non-esterified fatty acids (NEFA concentrations tended to be lower in the UF compared to the CF group (p = 0.058. Occupation ratios of Bacteroides and Clostridium cluster IV were significantly greater in the cecal flora of the UF group. Our results suggest that ume fiber possesses the fecal lipid excretion effects and feces bulking effects.

  8. Diversity in gut microflora of Helicoverpa armigera populations from different regions in relation to biological activity of Bacillus thuringiensis δ-endotoxin Cry1Ac.

    Science.gov (United States)

    Paramasiva, Inakarla; Shouche, Yogesh; Kulkarni, Girish Jayant; Krishnayya, Pulipaka Venkata; Akbar, Shaik Mohammed; Sharma, Hari Chand

    2014-12-01

    Transgenic crops expressing toxin proteins from Bacillus thuringiensis (Bt) have been deployed on a large scale for management of Helicoverpa armigera. Resistance to Bt toxins has been documented in several papers, and therefore, we examined the role of midgut microflora of H. armigera in its susceptibility to Bt toxins. The susceptibility of H. armigera to Bt toxin Cry1Ac was assessed using Log-dose-Probit analysis, and the microbial communities were identified by 16S rRNA sequencing. The H. armigera populations from nine locations harbored diverse microbial communities, and had some unique bacteria, suggesting a wide geographical variation in microbial community in the midgut of the pod borer larvae. Phylotypes belonging to 32 genera were identified in the H. armigera midgut in field populations from nine locations. Bacteria belonging to Enterobacteriaceae (Order Bacillales) were present in all the populations, and these may be the common members of the H. armigera larval midgut microflora. Presence and/or absence of certain species were linked to H. armigera susceptibility to Bt toxins, but there were no clear trends across locations. Variation in susceptibility of F1 neonates of H. armigera from different locations to the Bt toxin Cry1Ac was found to be 3.4-fold. These findings support the idea that insect migut microflora may influence the biological activity of Bt toxins.

  9. Ultrastructure of the digestive system and the fate of midgut during embryonic development in Porcellio scaber (Crustacea: Isopoda).

    Science.gov (United States)

    Strus, Jasna; Klepal, Waltraud; Repina, Janja; Tusek-Znidaric, Magda; Milatovic, Masa; Pipan, Ziva

    2008-07-01

    Microscopic anatomy of the digestive system in embryos and larvae of the terrestrial isopod crustacean Porcellio scaber was investigated by light bright field, fluorescence and electron microscopy. During marsupial ontogenetic development the event-dependent staging was used to discriminate the various embryonic stages. At the late embryo stage the differentiation of the ectodermal part of the gut into the complex filtering foregut and the hindgut with absorptive and transporting functions is accomplished. The gut of the marsupial manca larva is fully developed and similar to that of the adult. In early embryos the endodermal midgut gland primordia are filled with yolk and lipid globules. In late embryos the epithelium of paired midgut gland tubes is composed of two cell types; one of them exhibits orange autofluorescence. The endodermal cells located between the foregut and the midgut glands of late embryos form the prospective midgut. The cells have electron dense cytoplasm, abundant glycogen fields, endoplasmic reticulum, dictyosomes and numerous vesicles. In the adults the endodermal cells of the midgut remain only in the midgut gland ducts which connect the midgut glands and the foregut. Details of the cellular ultrastructure and morphogenesis of the ectodermal and endodermal parts of the digestive system during embryonic development of Porcellio scaber provide data for further phylogenetic and comparative studies in peracaridan crustaceans and other arthropods.

  10. Increased centrosome amplification in aged stem cells of the Drosophila midgut

    Energy Technology Data Exchange (ETDEWEB)

    Park, Joung-Sun; Pyo, Jung-Hoon; Na, Hyun-Jin; Jeon, Ho-Jun; Kim, Young-Shin [Department of Molecular Biology, Pusan National University, Busan 609-735 (Korea, Republic of); Arking, Robert, E-mail: aa2210@wayne.edu [Department of Biological Sciences, Wayne State University, Detroit, MI 48202 (United States); Yoo, Mi-Ae, E-mail: mayoo@pusan.ac.kr [Department of Molecular Biology, Pusan National University, Busan 609-735 (Korea, Republic of)

    2014-07-25

    Highlights: • Increased centrosome amplification in ISCs of aged Drosophila midguts. • Increased centrosome amplification in ISCs of oxidative stressed Drosophila midguts. • Increased centrosome amplification in ISCs by overexpression of PVR, EGFR, and AKT. • Supernumerary centrosomes can be responsible for abnormal ISC polyploid cells. • Supernumerary centrosomes can be a useful marker for aging stem cells. - Abstract: Age-related changes in long-lived tissue-resident stem cells may be tightly linked to aging and age-related diseases such as cancer. Centrosomes play key roles in cell proliferation, differentiation and migration. Supernumerary centrosomes are known to be an early event in tumorigenesis and senescence. However, the age-related changes of centrosome duplication in tissue-resident stem cells in vivo remain unknown. Here, using anti-γ-tubulin and anti-PH3, we analyzed mitotic intestinal stem cells with supernumerary centrosomes in the adult Drosophila midgut, which may be a versatile model system for stem cell biology. The results showed increased centrosome amplification in intestinal stem cells of aged and oxidatively stressed Drosophila midguts. Increased centrosome amplification was detected by overexpression of PVR, EGFR, and AKT in intestinal stem cells/enteroblasts, known to mimic age-related changes including hyperproliferation of intestinal stem cells and hyperplasia in the midgut. Our data show the first direct evidence for the age-related increase of centrosome amplification in intestinal stem cells and suggest that the Drosophila midgut is an excellent model for studying molecular mechanisms underlying centrosome amplification in aging adult stem cells in vivo.

  11. Recurrent Midgut Bleeding due to Jejunal Angioleiomyoma

    Directory of Open Access Journals (Sweden)

    Mahir Gachabayov

    2016-01-01

    Full Text Available Angioleiomyoma being a type of true smooth muscle gastrointestinal tumors can lead to serious life-threatening gastrointestinal bleeding. We report a case of 21-year-old male patient with recurrent midgut bleeding. Contrast-enhanced CT revealed highly vascular small bowel neoplasm. The patient underwent laparotomy with bowel resection and recovered uneventfully. Histopathology revealed jejunal angioleiomyoma.

  12. Recurrent Midgut Bleeding due to Jejunal Angioleiomyoma

    OpenAIRE

    2016-01-01

    Angioleiomyoma being a type of true smooth muscle gastrointestinal tumors can lead to serious life-threatening gastrointestinal bleeding. We report a case of 21-year-old male patient with recurrent midgut bleeding. Contrast-enhanced CT revealed highly vascular small bowel neoplasm. The patient underwent laparotomy with bowel resection and recovered uneventfully. Histopathology revealed jejunal angioleiomyoma.

  13. Proteomic analysis of the silkworm midgut during larval-pupal transition

    Directory of Open Access Journals (Sweden)

    M Kannan

    2016-06-01

    Full Text Available Metamorphosis is a key process in holometabolous insects since specific larval tissues, such as the midgut, the silk gland and the fat body, are remodeled into adult organs, or even disintegrate. However, the role of proteins and factors involved in the remodeling of these organs is still unclear. For this reason we undertook a proteomic study on the larval midgut of Bombyx mori to identify proteins whose expression significantly changes during larval-pupal metamorphosis and thus can have a role in this process. 2D-PAGE analysis showed upregulation or new expression of midgut proteins at 45 (pI 5.2, 29.8 (pI 6.11 and 20-kDa (pI 6-7 during the pupal stage. In addition zymogram analysis demonstrated the occurrence of protease activity at 37-kDa at the same developmental stage. By using MALDI-TOF-MS analysis the protein spots from 2D gel were identified as Hemolin, low molecular weight 30-kDa lipoproteins, HSP 20.8 and HSP 20.4, while the proteolytic band was identified as 37-kDa serine protease. The evidence herein presented suggests that the identified proteins could be involved in the immune protection during the remodeling of silkworm midgut that occurs at larval-pupal transition and represents a platform of knowledge necessary for future functional studies.

  14. Seasonality and Locality Affect the Diversity of Anopheles gambiae and Anopheles coluzzii Midgut Microbiota from Ghana.

    Science.gov (United States)

    Akorli, Jewelna; Gendrin, Mathilde; Pels, Nana Adjoa P; Yeboah-Manu, Dorothy; Christophides, George K; Wilson, Michael D

    2016-01-01

    Symbiotic bacteria can have important implications in the development and competence of disease vectors. In Anopheles mosquitoes, the composition of the midgut microbiota is largely influenced by the larval breeding site, but the exact factors shaping this composition are currently unknown. Here, we examined whether the proximity to urban areas and seasons have an impact on the midgut microbial community of the two major malaria vectors in Africa, An. coluzzii and An. gambiae. Larvae and pupae were collected from selected habitats in two districts of Ghana during the dry and rainy season periods. The midgut microbiota of adults that emerged from these collections was determined by 454-pyrosequencing of the 16S ribosomal DNA. We show that in both mosquito species, Shewanellaceae constituted on average of 54% and 73% of the midgut microbiota from each site in the dry and rainy season, respectively. Enterobacteriaceae was found in comparatively low abundance below 1% in 22/30 samples in the dry season, and in 25/38 samples in the rainy season. Our data indicate that seasonality and locality significantly affect both the diversity of microbiota and the relative abundance of bacterial families with a positive impact of dry season and peri-urban settings.

  15. Seasonality and Locality Affect the Diversity of Anopheles gambiae and Anopheles coluzzii Midgut Microbiota from Ghana

    Science.gov (United States)

    Gendrin, Mathilde; Pels, Nana Adjoa P.; Yeboah-Manu, Dorothy; Christophides, George K.; Wilson, Michael D.

    2016-01-01

    Symbiotic bacteria can have important implications in the development and competence of disease vectors. In Anopheles mosquitoes, the composition of the midgut microbiota is largely influenced by the larval breeding site, but the exact factors shaping this composition are currently unknown. Here, we examined whether the proximity to urban areas and seasons have an impact on the midgut microbial community of the two major malaria vectors in Africa, An. coluzzii and An. gambiae. Larvae and pupae were collected from selected habitats in two districts of Ghana during the dry and rainy season periods. The midgut microbiota of adults that emerged from these collections was determined by 454-pyrosequencing of the 16S ribosomal DNA. We show that in both mosquito species, Shewanellaceae constituted on average of 54% and 73% of the midgut microbiota from each site in the dry and rainy season, respectively. Enterobacteriaceae was found in comparatively low abundance below 1% in 22/30 samples in the dry season, and in 25/38 samples in the rainy season. Our data indicate that seasonality and locality significantly affect both the diversity of microbiota and the relative abundance of bacterial families with a positive impact of dry season and peri-urban settings. PMID:27322614

  16. Starches, resistant starches, the gut microflora and human health.

    Science.gov (United States)

    Bird, A R; Brown, I L; Topping, D L

    2000-03-01

    Starches are important as energy sources for humans and also for their interactions with the gut microflora throughout the digestive tact. Largely, those interactions promote human health. In the mouth, less gelatinised starches may lower risk of cariogensis. In the large bowel, starches which have escaped small intestinal digestion (resistant starch), together with proteins, other undigested carbohydrates and endogenous secretions are fermented by the resident microflora. The resulting short chain fatty acids contribute substantially to the normal physiological functions of the viscera. Specific types of resistant starch (e.g. the chemically modified starches used in the food industry) may be used to manipulate the gut bacteria and their products (including short chain fatty acids) so as to optimise health. In the upper gut, these starches may assist in the transport of probiotic organisms thus promoting the immune response and suppressing potential pathogens. However, it appears unlikely that current probiotic organisms can be used to modulate large bowel short chain fatty acids in adults although resistant starch and other prebiotics can do so. Suggestions that starch may exacerbate certain conditions (such as ulcerative colitis) through stimulating the growth of certain pathogenic organisms appear to be unfounded. Short chain fatty acids may modulate tissue levels and effects of growth factors in the gut and so modify gut development and risk of serious disease, including colo-rectal cancer. However, information on the relationship between starches and the microflora is relatively sparse and substantial opportunities exist both for basic research and food product development.

  17. Constitutive activation of the midgut response to Bacillus thuringiensis in Bt-resistant Spodoptera exigua.

    Directory of Open Access Journals (Sweden)

    Patricia Hernández-Martínez

    Full Text Available 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 involved in conferring resistance is of paramount importance. A colony of Spodoptera exigua (Lepidoptera: Noctuidae was selected for 15 years in the laboratory for resistance to Xentari™, a B. thuringiensis-based insecticide, reaching a final resistance level of greater than 1,000-fold. Around 600 midgut ESTs were analyzed by DNA-macroarray in order to find differences in midgut gene expression between susceptible and resistant insects. Among the differentially expressed genes, repat and arylphorin were identified and their increased expression was correlated with B. thuringiensis resistance. We also found overlap among genes that were constitutively over-expressed in resistant insects with genes that were up-regulated in susceptible insects after exposure to Xentari™, suggesting a permanent activation of the response to Xentari™ in resistant insects. Increased aminopeptidase activity in the lumen of resistant insects in the absence of exposure to Xentari™ corroborated the hypothesis of permanent activation of response genes. Increase in midgut proliferation has been proposed as a mechanism of response to pathogens in the adult from several insect species. Analysis of S. exigua larvae revealed that midgut proliferation was neither increased in resistant insects nor induced by exposure of susceptible larvae to Xentari™, suggesting that mechanisms other than midgut proliferation are involved in the response to B. thuringiensis by S. exigua larvae.

  18. Cytotoxic effects of neem oil in the midgut of the predator Ceraeochrysa claveri.

    Science.gov (United States)

    Scudeler, Elton Luiz; Garcia, Ana Silvia Gimenes; Padovani, Carlos Roberto; Pinheiro, Patricia Fernanda Felipe; dos Santos, Daniela Carvalho

    2016-01-01

    Studies of morphological and ultrastructural alterations in target organs have been useful for evaluating the sublethal effects of biopesticides regarded as safe for non-target organisms in ecotoxicological analyses. One of the most widely used biopesticides is neem oil, and its safety and compatibility with natural enemies have been further clarified through bioassays performed to analyze the effects of indirect exposure by the intake of poisoned prey. Thus, this study examined the cellular response of midgut epithelial cells of the adult lacewing, Ceraeochrysa claveri, to neem oil exposure via intake of neem oil-contaminated prey during the larval stage. C. claveri larvae were fed Diatraea saccharalis eggs treated with neem oil at concentrations of 0.5%, 1% and 2% throughout the larval stage. The adult females obtained from these treatments were used at two ages (newly emerged and at the start of oviposition) in morphological and ultrastructural analyses. Neem oil was found to cause pronounced cytotoxic effects in the adult midgut, such as cell dilation, emission of cytoplasmic protrusions, cell lysis, loss of integrity of the cell cortex, dilation of cisternae of the rough endoplasmic reticulum, swollen mitochondria, vesiculated appearance of the Golgi complex and dilated invaginations of the basal labyrinth. Epithelial cells responded to those injuries with various cytoprotective and detoxification mechanisms, including increases in cell proliferation, the number of calcium-containing cytoplasmic granules, and HSP 70 expression, autophagic processes and the development of smooth endoplasmic reticulum, but these mechanisms were insufficient for recovery from all of the cellular damage to the midgut. This study demonstrates that neem oil exposure impairs the midgut by causing sublethal effects that may affect the physiological functions of this organ, indicating the importance of studies of different life stages of this species and similar species to evaluate the

  19. Microflora en semillas de frijol

    Directory of Open Access Journals (Sweden)

    Jos\\u00E9 B. Membre\\u00F1o

    2001-01-01

    Full Text Available Microflora en semillas de frijol (Phaseolus vulgaris L.. Se estudió la microflora bacteriana presente en semillas de frijol y su relación con Xanthomonas campestris pv. phaseoli (Xcp, en 118 genotipos procedentes de VIDAC-98, INTA- Nicaragua, TARS-USDA e Isabela-P.R. Se utilizaron cinco métodos de aislamiento: semilla desinfectada con hipoclorito de sodio, semilla en caldo nutritivo refrigerada por una hora, dispersión de 0,1 ml de suspensión de semillas en medio sólido, siembra líquida de 1 ml de suspensión y semilla en caldo nutritivo, agitado y refrigerado por 24 horas. Se aislaron 104 colonias amarillas de 41 genotipos. Treinta y seis colonias fueron KOH positivo (Gram negativo, 68 negativo (Gram positivo y 34 hidrolizaron almidón. Las colonias de pigmentación amarilla resultaron no patogénicas bajo condiciones de invernadero. Estas se identificaron con el sistema BIOLOG como: Pantoea agglomerans (25, Xanthomonas campestris (2, Enterobacter agglomerans (2, Sphingomonas paucimobilis (2, Pseudomonas fluorescens y Flavimonas oryzihabitans. En adición, los genotipos portaron colonias con pigmentación distinta a la amarilla. En las pruebas de antagonismo se identificaron colonias con actividad de deoxyribonucleasa y de antibiosis a Xcp. De éstas, 15 colonias inhibieron a Xcp significativamente. Se identificaron los hongos Rhizoctonia solani, Penicillium spp., Fusarium spp., Aspergillus flavus, Rhizopus nigricans y Macrophomina phaseolina en un 52,9 % del total de genotipos evaluados

  20. Changes in oral microflora after full-mouth tooth extraction : A prospective cohort study

    NARCIS (Netherlands)

    de Waal, Yvonne C M; Winkel, Edwin G; Raangs, Gerwin C; van der Vusse, Marleen L; Rossen, John W A; van Winkelhoff, Arie Jan

    2014-01-01

    AIM: The aim of the study was to evaluate the effect of full-mouth tooth extraction on the oral microflora, with emphasis on the presence and load of Aggregatibacter actinomycetemcomitans and Porphyromonas gingivalis. MATERIAL AND METHODS: Adult patients (n = 30), with moderate to advanced periodont

  1. An atypical clinical presentation of acute appendicitis in a young man with midgut malrotation

    Energy Technology Data Exchange (ETDEWEB)

    Pinto, Antonio [Dipartimento Biomedico di Medicina Interna e Specialistica, University of Palermo, Piazza delle Cliniche no 2, 90127 Palermo (Italy)]. E-mail: pinto@neomedia.it; Di Raimondo, Domenico [Dipartimento Biomedico di Medicina Interna e Specialistica, University of Palermo, Piazza delle Cliniche no 2, 90127 Palermo (Italy); Tuttolomondo, Antonino [Dipartimento Biomedico di Medicina Interna e Specialistica, University of Palermo, Piazza delle Cliniche no 2, 90127 Palermo (Italy); Fernandez, Paola [Dipartimento Biomedico di Medicina Interna e Specialistica, University of Palermo, Piazza delle Cliniche no 2, 90127 Palermo (Italy); Caronia, Aurelio [Dipartimento di Biotecnologie Mediche - Sezione di Radiologia, University of Palermo, Via del Vespro no 129, 90127 Palermo (Italy); Lagalla, Roberto [Dipartimento di Biotecnologie Mediche - Sezione di Radiologia, University of Palermo, Via del Vespro no 129, 90127 Palermo (Italy); Arnao, Valentina [Dipartimento Biomedico di Medicina Interna e Specialistica, University of Palermo, Piazza delle Cliniche no 2, 90127 Palermo (Italy); Law, Robert L. [Department of Radiology, Frenchay Hospital, Bristol (United Kingdom); Licata, Giuseppe [Dipartimento Biomedico di Medicina Interna e Specialistica, University of Palermo, Piazza delle Cliniche no 2, 90127 Palermo (Italy)

    2007-05-15

    Midgut malrotation occurs as a result of failure in normal intestinal rotation and fixation during early pregnancy. Pathological conditions reported in the literature involving midgut malrotation predominantly relate to infants and children. In adults malrotation is often revealed as an incidental finding on computed tomography (CT), or the associated altered anatomy can be the cause of atypical clinical symptoms of relatively common intestinal disorders. An unusual presentation of acute appendicitis, with fever and recurrent pain in left iliac fossa is reported. Underlying intestinal malrotation delayed the correct clinical diagnosis of acute appendicitis. It was not until a CT scan was performed that a malrotation was identified. The predominant appearances of malrotation are the siting of the ascending colon, caecum (and appendix) in the left side of the abdomen and the right-sided placement of the duodenojejunal junction.

  2. Particularlies normal microflora of the human

    Directory of Open Access Journals (Sweden)

    T. V. Sklyar

    2005-02-01

    Full Text Available Nowdays it marks the constant growth of diseases connected to changes of biological balance between macroorganism and various microbial populations of its organs and systems which formed during evolution. The literary data and experimental data of artors are generalised in this article. They concern structure microflora of human organism, factors influencing process of its formation, meaning normal microflora for functioning organism as a whole, and for systems and organs

  3. Bacterial Communities and Midgut Microbiota Associated with Mosquito Populations from Waste Tires in East-Central Illinois.

    Science.gov (United States)

    Kim, Chang-Hyun; Lampman, Richard L; Muturi, Ephantus J

    2015-01-01

    Mosquito-microbe interactions tend to influence larval nutrition, immunity, and development, as well as fitness and vectorial capacity of adults. Understanding the role of different bacterial species not only improves our knowledge of the physiological and ecological consequences of these interactions, but also provides the basis for developing novel strategies for controlling mosquito-borne diseases. We used culture-dependent and culture-independent techniques to characterize the bacterial composition and abundance in water and midgut samples of larval and adult females of Aedes japonicus (Theobald), Aedes triseriatus (Say), and Culex restuans (Theobald) collected from waste tires at two wooded study sites in Urbana, IL. The phylum-specific real-time quantitative polymerase chain reaction assay revealed a higher proportion of Actinobacteria and a lower proportion of gamma-Proteobacteria and Bacteroidetes in water samples and larval midguts compared to adult female midguts. Only 15 of the 57 bacterial species isolated in this study occurred in both study sites. The number of bacterial species was highest in water samples (28 species from Trelease Woods; 25 species from South Farms), intermediate in larval midguts (13 species from Ae. japonicus; 12 species from Ae. triseriatus; 8 species from Cx. restuans), and lowest in adult female midguts (2 species from Ae. japonicus; 3 species from Ae. triseriatus). These findings suggest that the composition and richness of bacterial communities varies both between habitats and among mosquito species and that the reduction in bacteria diversity during metamorphosis is more evident among bacteria detected using the culture-dependent method.

  4. Proteomic Analysis of Larval Midgut from the Silkworm (Bombyx mori

    Directory of Open Access Journals (Sweden)

    Sai Zhang

    2011-01-01

    Full Text Available The midgut is the major organ for food digestion, nutrient absorption and also a barrier for foreign substance. The 5th-instar larval stage of silkworm is very important for larval growth, development, and silk production. In the present study, we used 2-DE and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS to analyze the midgut proteins from the 5th-instar larvae as well as the midgut proteins under starvation condition. A total of 96 proteins were identified in this study; and among them, 69 proteins were observed in midgut for the first time. We also found that the silkworm larval midgut responded to starvation by producing a 10 kDa heat shock protein and a diapause hormone precursor.

  5. Imidacloprid impairs the post-embryonic development of the midgut in the yellow fever mosquito Stegomyia aegypti (=Aedes aegypti).

    Science.gov (United States)

    Fernandes, K M; Gonzaga, W G; Pascini, T V; Miranda, F R; Tomé, H V V; Serrão, J E; Martins, G F

    2015-09-01

    The mosquito Stegomyia aegypti (=Aedes aegypti) (Diptera: Culicidae) is a vector for the dengue and yellow fever viruses. As blood digestion occurs in the midgut, this organ constitutes the route of entry of many pathogens. The effects of the insecticide imidacloprid on the survival of St. aegypti were investigated and the sub-lethal effects of the insecticide on midgut development were determined. Third instar larvae were exposed to different concentrations of imidacloprid (0.15, 1.5, 3.0, 6.0 and 15.0 p.p.m.) and survival was monitored every 24 h for 10 days. Midguts from imidacloprid-treated insects at different stages of development were dissected and processed for analyses by transmission electron microscopy, immunofluorescence microscopy and terminal deoxynucleotidyl transferase dUTP nick-end labelling (TUNEL) assays. Imidacloprid concentrations of 3.0 and 15.0 p.p.m. were found to affect midgut development similarly. Digestive cells of the fourth instar larvae (L4) midgut exposed to imidacloprid had more multilamellar bodies, abundantly found in the cell apex, and more electron-lucent vacuoles in the basal region compared with those from untreated insects. Moreover, imidacloprid interfered with the differentiation of regenerative cells, dramatically reducing the number of digestive and endocrine cells and leading to malformation of the midgut epithelium in adults. The data demonstrate that imidacloprid can reduce the survival of mosquitoes and thus indicate its potentially high efficacy in the control of St. aegypti populations.

  6. Desenvolvimento pós-embrionário do intestino médio de Dermatobia hominis (Linnaeus Jr. (Diptera, Cuterebridae Post-embryonic development of Dermatobia hominis (Linnaeus Jr. (Diptera, Cuterebridae midgut

    Directory of Open Access Journals (Sweden)

    Edy de Lello

    2001-03-01

    Full Text Available Dermatobia hominis (Linnaeus, 1781 midgut is internally lined by an epithelium of polytenic cells, some low others prismatic with well developed brush border. Their apical portion are enlarged by secretory vesicles, forming button-like structures that are pinched off to the lumen, some accompained by the nucleus characterizing apocrine and holocrine secretions. This epithelium is gradually renewed by small, non polytenic regenerative cells, found scattered at its basal portion. At the end of the third instar the metamorphosis begins. The epithelial cells present signs of degeneration and at the first day of pupation the regenerative cells increase in number. By the 5th day of pupation these regenerative cells, besides being increased in number, differentiate themselves into two layers: one similar to the dense conective tissue that sustainning the larval epithelium is pinched off to the midgut lumen forming the "yellow bodies"; the other, develops right under it as the imaginal epitelium. The disorganized muscles bundles of the midgut wall, are invaded by phagocytes. At the end of pupation the midgut has a low prismatic epithelium with brush-border. In the adult, the torax portion of the midgut has prismatic homogeneously basophilic epithelium while in the abdominal portion the epithelium is made of high prismatic cells full of small vacuoles. The larval midgut epithelium suffers programmed cell death non compatible with apoptose. During the metamorphosis the midgut lenght diminishes from 31mm in the larva to 14mm in the adult.

  7. Production and characterization of monoclonal antibodies against midgut of ixodid tick, Haemaphysalis longicornis.

    Science.gov (United States)

    Nakajima, Mie; Kodama, Michi; Yanase, Haruko; Iwanaga, Toshihiko; Mulenga, Albert; Ohashi, Kazuhiko; Onuma, Misao

    2003-08-14

    There are concerted efforts toward development of tick vaccines to replace current chemical control strategies that have serious limitations [Parasitologia 32 (1990) 145; Infectious Disease Clinics of North America (1999) 209-226]. In this study, monoclonal antibodies (mAbs) specific to Haemaphysalis longicornis midgut proteins were produced and characterized. Eight antibody-secreting hybridomas were cloned and the mAbs typed as IgG1, IgG2a and IgG2b. On immunoblots, all mAbs reacted with a midgut protein band of about 76 kDa. All mAbs uniformly immunogold-stained the surface or epithelial layers of H. longicornis midgut and endosomes. Adult ticks (50%) that fed on an ascitic mouse producing the IgGs developed a red coloration and did not oviposit. As such, the 76 kDa protein that reacted with the mAbs could, therefore, be a potential candidate for tick vaccine development.

  8. Plasma membranes from insect midgut cells

    Directory of Open Access Journals (Sweden)

    Walter R. Terra

    2006-06-01

    Full Text Available Plasma membranes from insect midgut cells are separated into apical and basolateral domains. The apical domain is usually modified into microvilli with a molecular structure similar to other animals. Nevertheless, the microvillar structure should differ in some insects to permit the traffic inside them of secretory vesicles that may budd laterally or pinch-off from the tips of microvilli. Other microvillar modifications are associated with proton-pumping or with the interplay with an ensheathing lipid membrane (the perimicrovilllar membrane observed in the midgut cells of hemipterans (aphids and bugs. The perimicrovillar membranes are thought to be involved in amino acid absorption from diluted diets. The microvillar and perimicrovillar membranes have densities (and protein content that depend on the insect taxon. The role played by the microvillar and perimicrovillar proteins in insect midgut physiology is reviewed here trying to provide a coherent picture of data and highlighting further research areas.As membranas plasmáticas das células intestinais dos insetos apresentam um domínio apical e outro basal. O domínio apical é geralmente modificado em microvilosidades com organização molecular similar a de outros animais, embora possam diferir naqueles insetos que apresentam vesículas secretoras em trânsito que brotam lateralmente ou destacam-se das extremidades das microvilosidades. Outras modificações microvilares estão associadas a bombeamento de prótons ou a interrelações com uma membrana lipídica (a membrana perimicrovilar que reveste as microvilosidades de células intestinais de hemípteros (pulgões e percevejos. Admite-se que as membranas perimicrovilares estejam envolvidas na absorção de aminoácidos a partir de dietas diluídas. As membranas microvilares e perimicrovilares tem densidades distintas (e conteúdo protéico que dependem do táxon do inseto. O papel desempenhado pelas proteínas microvilares e

  9. Microflora analysis of a child with severe combined immune deficiency

    Science.gov (United States)

    Taylor, G. R.; Kropp, K. D.; Molina, T. C.

    1978-01-01

    The paper presents a microflora analysis of a 5-year-old male child with severe combined immune deficiency who was delivered by Caesarean section and continuously maintained in an isolator. Despite precautions, it was found that the child had come in contact with at least 54 different microbial contaminants. While his skin autoflora was similar to that of a reference group of healthy male adults in numbers of different species and the number of viable cells present per square centimeter of surface area, the subject's autoflora differed from the reference group in that significantly fewer anaerobic species were recovered from the patient's mouth and feces. It is suggested that the child's remaining disease free shows that the reported bacteria are noninvasive or that the unaffected components of the child's immune defense mechanisms are important.

  10. Oral microflora in children with hematologic malignancies

    OpenAIRE

    M. F. Vecherkovskaya; G. V. Tets; B. V. Afanasiev; V. V. Tets

    2015-01-01

    The goal was a comprehensive study of oral microflora in healthy children and those with hematologic malignancies, based on the analysis of mixed microbial biofilms composition, isolation and identification of new previously unknown microorganisms. The material was obtained in children with hematological diseases in remission, 2–10 years aged, and for the control group from St. Petersburg schoolchildren and in kindergartens. We used microbiological, biochemical and molecular genetic methods, ...

  11. Laminin and the malaria parasite's journey through the mosquito midgut.

    Science.gov (United States)

    Arrighi, Romanico B G; Lycett, Gareth; Mahairaki, Vassiliki; Siden-Kiamos, Inga; Louis, Christos

    2005-07-01

    During the invasion of the mosquito midgut epithelium, Plasmodium ookinetes come to rest on the basal lamina, where they transform into the sporozoite-producing oocysts. Laminin, one of the basal lamina's major components, has previously been shown to bind several surface proteins of Plasmodium ookinetes. Here, using the recently developed RNAi technique in mosquitoes, we used a specific dsRNA construct targeted against the LANB2 gene (laminin gamma1) of Anopheles gambiae to reduce its mRNA levels, leading to a substantial reduction in the number of successfully developed oocysts in the mosquito midgut. Moreover, this molecular relationship is corroborated by the intimate association of developing P. berghei parasites and laminin in the gut, as observed using confocal microscopy. Our data support the notion of laminin playing a functional role in the development of the malaria parasite within the mosquito midgut.

  12. Impact of salinomycin on the intestinal microflora of broiler chickens

    DEFF Research Database (Denmark)

    Johansen, Charlotte; Friis-Holm, Lotte Bjerrum; Pedersen, Karl

    2007-01-01

    jejuni infection and on the composition of the caecal microflora in broiler chickens. Methods: An experimental infection study was carried out in isolators and the intestinal microflora was analyzed using quantitative cultivation, denaturant gradient gel electrophoresis (DGGE), cloning and sequencing....... Results: We found no effect of salinomycin on C. jejuni but salinomycin significantly affected the composition of the microflora. In addition, salinomycin significantly reduced the prevalence of Clostridium perfringens and we observed a significant increase (62%) in the mean body weight of salinomycin...

  13. Schinus terebinthifolius Leaf Extract Causes Midgut Damage, Interfering with Survival and Development of Aedes aegypti Larvae.

    Directory of Open Access Journals (Sweden)

    Thamara Figueiredo Procópio

    Full Text Available In this study, a leaf extract from Schinus terebinthifolius was evaluated for effects on survival, development, and midgut of A. aegypti fourth instar larvae (L4, as well as for toxic effect on Artemia salina. Leaf extract was obtained using 0.15 M NaCl and evaluated for phytochemical composition and lectin activity. Early L4 larvae were incubated with the extract (0.3-1.35%, w/v for 8 days, in presence or absence of food. Polymeric proanthocyanidins, hydrolysable tannins, heterosid and aglycone flavonoids, cinnamic acid derivatives, traces of steroids, and lectin activity were detected in the extract, which killed the larvae at an LC50 of 0.62% (unfed larvae and 1.03% (fed larvae. Further, the larvae incubated with the extract reacted by eliminating the gut content. No larvae reached the pupal stage in treatments at concentrations between 0.5% and 1.35%, while in the control (fed larvae, 61.7% of individuals emerged as adults. The extract (1.0% promoted intense disorganization of larval midgut epithelium, including deformation and hypertrophy of cells, disruption of microvilli, and vacuolization of cytoplasms, affecting digestive, enteroendocrine, regenerative, and proliferating cells. In addition, cells with fragmented DNA were observed. Separation of extract components by solid phase extraction revealed that cinnamic acid derivatives and flavonoids are involved in larvicidal effect of the extract, being the first most efficient in a short time after larvae treatment. The lectin present in the extract was isolated, but did not show deleterious effects on larvae. The extract and cinnamic acid derivatives were toxic to A. salina nauplii, while the flavonoids showed low toxicity. S. terebinthifolius leaf extract caused damage to the midgut of A. aegypti larvae, interfering with survival and development. The larvicidal effect of the extract can be attributed to cinnamic acid derivatives and flavonoids. The data obtained using A. salina indicates

  14. Schinus terebinthifolius Leaf Extract Causes Midgut Damage, Interfering with Survival and Development of Aedes aegypti Larvae

    Science.gov (United States)

    Procópio, Thamara Figueiredo; Fernandes, Kenner Morais; Pontual, Emmanuel Viana; Ximenes, Rafael Matos; de Oliveira, Aline Rafaella Cardoso; Souza, Carolina de Santana; Melo, Ana Maria Mendonça de Albuquerque; Navarro, Daniela Maria do Amaral Ferraz; Paiva, Patrícia Maria Guedes; Martins, Gustavo Ferreira; Napoleão, Thiago Henrique

    2015-01-01

    In this study, a leaf extract from Schinus terebinthifolius was evaluated for effects on survival, development, and midgut of A. aegypti fourth instar larvae (L4), as well as for toxic effect on Artemia salina. Leaf extract was obtained using 0.15 M NaCl and evaluated for phytochemical composition and lectin activity. Early L4 larvae were incubated with the extract (0.3–1.35%, w/v) for 8 days, in presence or absence of food. Polymeric proanthocyanidins, hydrolysable tannins, heterosid and aglycone flavonoids, cinnamic acid derivatives, traces of steroids, and lectin activity were detected in the extract, which killed the larvae at an LC50 of 0.62% (unfed larvae) and 1.03% (fed larvae). Further, the larvae incubated with the extract reacted by eliminating the gut content. No larvae reached the pupal stage in treatments at concentrations between 0.5% and 1.35%, while in the control (fed larvae), 61.7% of individuals emerged as adults. The extract (1.0%) promoted intense disorganization of larval midgut epithelium, including deformation and hypertrophy of cells, disruption of microvilli, and vacuolization of cytoplasms, affecting digestive, enteroendocrine, regenerative, and proliferating cells. In addition, cells with fragmented DNA were observed. Separation of extract components by solid phase extraction revealed that cinnamic acid derivatives and flavonoids are involved in larvicidal effect of the extract, being the first most efficient in a short time after larvae treatment. The lectin present in the extract was isolated, but did not show deleterious effects on larvae. The extract and cinnamic acid derivatives were toxic to A. salina nauplii, while the flavonoids showed low toxicity. S. terebinthifolius leaf extract caused damage to the midgut of A. aegypti larvae, interfering with survival and development. The larvicidal effect of the extract can be attributed to cinnamic acid derivatives and flavonoids. The data obtained using A. salina indicates that caution

  15. Origin and dynamic lineage characteristics of the developing Drosophila midgut stem cells.

    Science.gov (United States)

    Takashima, Shigeo; Aghajanian, Patrick; Younossi-Hartenstein, Amelia; Hartenstein, Volker

    2016-08-15

    Proliferating intestinal stem cells (ISCs) generate all cell types of the Drosophila midgut, including enterocytes, endocrine cells, and gland cells (e.g., copper cells), throughout the lifetime of the animal. Among the signaling mechanisms controlling the balance between ISC self-renewal and the production of different cell types, Notch (N) plays a pivotal role. In this paper we investigated the emergence of ISCs during metamorphosis and the role of N in this process. Precursors of the Drosophila adult intestinal stem cells (pISCs) can be first detected within the pupal midgut during the first hours after onset of metamorphosis as motile mesenchymal cells. pISCs perform 2-3 rounds of parasynchronous divisions. The first mitosis yields only an increase in pISC number. During the following rounds of mitosis, dividing pISCs give rise to more pISCs, as well as the endocrine cells that populate the midgut of the eclosing fly. Enterocytes do not appear among the pISC progeny until around the time of eclosion. The "proendocrine" gene prospero (pros), expressed from mid-pupal stages onward in pISCs, is responsible to advance the endocrine fate in these cells; following removal of pros, pISCs continue to proliferate, but endocrine cells do not form. Conversely, the onset of N activity that occurs around the stage when pros comes on restricts pros expression among pISCs. Loss of N abrogates proliferation and switches on an endocrine fate among all pISCs. Our results suggest that a switch depending on the activity of N and pros acts at the level of the pISC to decide between continued proliferation and endocrine differentiation.

  16. Plasmodium ookinetes coopt mammalian plasminogen to invade the mosquito midgut

    DEFF Research Database (Denmark)

    Ghosh, Anil K; Coppens, Isabelle; Gårdsvoll, Henrik;

    2011-01-01

    inhibits oocyst development of both Plasmodium berghei and Plasmodium falciparum, suggesting that enolase may act as an invasion ligand. Importantly, we demonstrate that surface enolase captures plasminogen from the mammalian blood meal via its lysine motif (DKSLVK) and that this interaction is essential...... for ookinete invasion. The results support the hypothesis that enolase on the surface of Plasmodium ookinetes plays a dual role in midgut invasion: by acting as a ligand that interacts with the midgut epithelium and, further, by capturing plasminogen, whose conversion to active plasmin promotes the invasion...

  17. Analysis of two novel midgut-specific promoters driving transgene expression in Anopheles stephensi mosquitoes.

    Directory of Open Access Journals (Sweden)

    Tony Nolan

    Full Text Available BACKGROUND: Tissue-specific promoters controlling the expression of transgenes in Anopheles mosquitoes represent a valuable tool both for studying the interaction between these malaria vectors and the Plasmodium parasites they transmit and for novel malaria control strategies based on developing Plasmodium-refractory mosquitoes by expressing anti-parasitic genes. With this aim we have studied the promoter regions of two genes from the most important malaria vector, Anopheles gambiae, whose expression is strongly induced upon blood feeding. RESULTS: We analysed the A. gambiae Antryp1 and G12 genes, which we have shown to be midgut-specific and maximally expressed at 24 hours post-bloodmeal (PBM. Antryp1, required for bloodmeal digestion, encodes one member of a family of 7 trypsin genes. The G12 gene, of unknown function, was previously identified in our laboratory in a screen for genes induced in response to a bloodmeal. We fused 1.1 kb of the upstream regions containing the putative promoter of these genes to reporter genes and transformed these into the Indian malaria vector A. stephensi to see if we could recapitulate the expression pattern of the endogenous genes. Both the Antryp1 and G12 upstream regions were able to drive female-predominant, midgut-specific expression in transgenic mosquitoes. Expression of the Antryp1-driven reporter in transgenic A. stephensi lines was low, undetectable by northern blot analysis, and failed to fully match the induction kinetics of the endogenous Antryp1 gene in A. gambiae. This incomplete conservation of expression suggests either subtle differences in the transcriptional machinery between A. stephensi and A. gambiae or that the upstream region chosen lacked all the control elements. In contrast, the G12 upstream region was able to faithfully reproduce the expression profile of the endogenous A. gambiae gene, showing female midgut specificity in the adult mosquito and massive induction PBM, peaking at 24

  18. Model intestinal microflora in computer simulation : A simulation and modeling package for host-microflora interactions

    NARCIS (Netherlands)

    Wilkinson, Michael H.F.

    2002-01-01

    The ecology of the human intestinal microflora and its interaction with the host are poorly understood. Though more and more data are being acquired, in part using modern molecular methods, development of a quantitative theory has not kept pace with this increase in observing power. This is in part

  19. 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; Ercolini, Danilo; Pennacchio, Francesco

    2016-08-23

    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.

  20. Complications of midgut carcinoid tumors and carcinoid syndrome

    NARCIS (Netherlands)

    van der Horst-Schrivers, Anouk N A; Wymenga, A N Machteld; Links, Thera P; Willemse, Pax H B; Kema, Ido P; de Vries, Elisabeth G E

    2004-01-01

    The carcinoid syndrome, associated with carcinoid tumors of the midgut, consists of symptoms such as diarrhea, flushing, wheezing and cardiovascular symptoms. This review focuses on these symptoms and discusses therapeutic options. The symptoms are caused by the secretion of biogenic amines, polypep

  1. Salivary microflora and mode of delivery

    DEFF Research Database (Denmark)

    Boustedt, Katarina; Roswall, Josefine; Dahlén, Gunnar

    2015-01-01

    BACKGROUND: Previous cross-sectional studies have suggested that the mode of delivery can influence the composition of oral microflora. The aim of this prospective study was to compare the salivary colonization in vaginally delivered children with children delivered by Caesarian section (C...... months. A saliva sample from the mothers was obtained 6 months after delivery. The parents were asked to complete a questionnaire on socioeconomic factors, lifestyle, and hygiene at baseline and throughout the study period. All samples were analyzed with 13 pre-determined bacterial probes using...

  2. Midgut proteins released by microapocrine secretion in Spodoptera frugiperda.

    Science.gov (United States)

    Silva, Walciane; Cardoso, Christiane; Ribeiro, Alberto F; Terra, Walter R; Ferreira, Clélia

    2013-01-01

    Microapocrine vesicles bud from the lepidopteran midgut microvilli as double membrane vesicles. To identify the proteins secreted by this process, antibodies raised against isolated microapocrine vesicles from Spodoptera frugiperda were used for screening a midgut cDNA expression library. Positive clones were sequenced, assembled and N blasted against S. frugiperda sequences obtained by pyrosequencing midgut mRNA. This procedure led to the extension of microapocrine sequences that were annotated. A similar procedure was used to identify midgut microvillar proteins that necessarily are part of the microapocrine vesicle. Forty-eight proteins were associated with microvillar membranes. They pertain to 8 functional groups: digestive enzymes, peritrophic membrane, protection, transporters, receptors, secretory machinery, cytoskeleton and signaling, and unknown. Twenty-eight proteins are putatively secreted by microapocrine secretion. Most of them are digestive enzymes, but the list also includes proteins involved in protection and in peritrophic membrane formation. Among the identified digestive enzymes, aminopeptidases are typically microvillar and group into the classes 1, 2, 3, 5, and 6. There are two amylases secreted by microapocrine secretion: one is a digestive enzyme and the other is a transporter-like amylase with no clear function. One lipase has a predicted transmembrane loop, whereas the others are supposed to be secreted by microapocrine secretion and be digestive. Trypsin is membrane bound and is delivered by microapocrine secretion, but has no predicted features to bind membranes. It may remain bound through the signal peptide till be delivered into the midgut lumen. Proteins supposed to be involved in the microapocrine secretory machinery were: calmodulin, annexin, myosin 7a, and gelsolin 1. Their putative roles are discussed, but more research is necessary to settle this subject.

  3. Spatial and sex-specific dissection of the Anopheles gambiae midgut transcriptome

    Directory of Open Access Journals (Sweden)

    Mahairaki Vassiliki

    2007-01-01

    Full Text Available Abstract Background The midgut of hematophagous insects, such as disease transmitting mosquitoes, carries out a variety of essential functions that mostly relate to blood feeding. The midgut of the female malaria vector mosquito Anopheles gambiae is a major site of interactions between the parasite and the vector. Distinct compartments and cell types of the midgut tissue carry out specific functions and vector borne pathogens interact and infect different parts of the midgut. Results A microarray based global gene expression approach was used to compare transcript abundance in the four major female midgut compartments (cardia, anterior, anterior part of posterior and posterior part of posterior midgut and between the male and female Anopheles gambiae midgut. Major differences between the female and male midgut gene expression relate to digestive processes and immunity. Each compartment has a distinct gene function profile with the posterior midgut expressing digestive enzyme genes and the cardia and anterior midgut expressing high levels of antimicrobial peptide and other immune gene transcripts. Interestingly, the cardia expressed several known anti-Plasmodium factors. A parallel peptidomic analysis of the cardia identified known mosquito antimicrobial peptides as well as several putative short secreted peptides that are likely to represent novel antimicrobial factors. Conclusion The A. gambiae sex specific midgut and female midgut compartment specific transcriptomes correlates with their known functions. The significantly greater functional diversity of the female midgut relate to hematophagy that is associated with digestion and nutrition uptake as well as exposes it to a variety of pathogens, and promotes growth of its endogenous microbial flora. The strikingly high proportion of immunity related factors in the cardia tissue most likely serves the function to increase sterility of ingested sugar and blood. A detailed characterization of the

  4. Oral microflora in children with hematologic malignancies

    Directory of Open Access Journals (Sweden)

    M. F. Vecherkovskaya

    2015-06-01

    Full Text Available The goal was a comprehensive study of oral microflora in healthy children and those with hematologic malignancies, based on the analysis of mixed microbial biofilms composition, isolation and identification of new previously unknown microorganisms. The material was obtained in children with hematological diseases in remission, 2–10 years aged, and for the control group from St. Petersburg schoolchildren and in kindergartens. We used microbiological, biochemical and molecular genetic methods, including electron microscopy, proteomic analysis, sequencing and complete genome annotation. Microorganisms of 23 genera isolated as pure cultures and identified by biochemical activity from mixed microbial biofilm derived from saliva of healthy and sick children. In microflora of children with hematologic malignancies a previously unknown type of streptococci with a large number of antibiotic resistance genes was revealed. Differences in oral microbiota composition of healthy children and children with hematological diseases in remission were revealed. The microbiota of children with hematologic malignancies contains more genes controlling antibiotic resistance. Also, it was observed previously unknown bacterium of the genus Streptococcus.

  5. Fermentative hydrogen production by diverse microflora

    Energy Technology Data Exchange (ETDEWEB)

    Baghchehsaraee, Bita; Nakhla, George; Karamanev, Dimitre; Margaritis, Argyrios [Department of Chemical and Biochemical Engineering, The University of Western Ontario, London, Ontario N6A 5B9 (Canada)

    2010-05-15

    In this study, hydrogen production with activated sludge, a diverse bacterial source has been investigated and compared to microflora from anaerobic digester sludge, which is less diverse. Batch experiments were conducted at mesophilic (37 C) and thermophilic (55 C) temperatures. The hydrogen production yields with activated sludge at 37 C and 55 C were 0.56 and 1.32 mol H{sub 2}/mol glucose consumed, respectively. While with anaerobically digested sludge hydrogen yield was 2.18 mol H{sub 2}/mol glucose consumed at 37 C and 1.25 mol H{sub 2}/mol glucose consumed at 55 C. The results of repeated batch experiments for 615 h resulted in average yields of 1.21 {+-} 0.62 and 1.40 {+-} 0.16 mol H{sub 2}/mol glucose consumed for activated sludge and anaerobic sludge, respectively. The hydrogen production with activated sludge was not stable during the repeated batches and the fluctuation in hydrogen production was attributed to formation of lactic acid as the predominant metabolite in some batches. The presence of lactic acid bacteria in microflora was confirmed by PCR-DGGE. (author)

  6. Digestive and regenerative cells in the midgut of haploid and diploid males of the stingless bee Melipona quadrifasciata anthidioides (Hymenoptera: Apidae

    Directory of Open Access Journals (Sweden)

    Kenner M. Fernandes

    2012-10-01

    Full Text Available In eusocial bees, workers and queens are diploid (2n, whereas males are haploid (n. However, in some species, including the stingless bee Melipona quadrifasciata anthidioides Lepeletier, 1836, 2n males arise from fertilized eggs resulting from the crossing between a queen and her brother. In the present study, we provide a comparative analysis of the digestive and regenerative cells in n and 2n pupae and adult males of M. quadrifasciata anthidioides. In n and 2n pupae and adult males, the number of regenerative cells/nest was similar. In n and 2n pupae, the mean number of digestive cells/midgut area was 2076 ± 0.60, whereas in adults it was 1234 ± 1.42 digestive cells/midgut area. The nuclear area of the digestive cells was also similar in both n and 2n adult males (~154 µm² and smaller in pupae (~91 µm²; this variation might be a result of DNA amplification in digestive cells during bee development. The results from our current study provide further understanding of the morphological and physiological aspects of the digestive tract of bees and show that the ploidy difference between n and 2n male stages does not affect the number of digestive and regenerative cells in the midgut of M. quadrifasciata anthidioides.

  7. Side Effects of Neem Oil on the Midgut Endocrine Cells of the Green Lacewing Ceraeochrysa claveri (Navás) (Neuroptera: Chrysopidae).

    Science.gov (United States)

    Scudeler, E L; Santos, D C

    2014-04-01

    We described the ultrastructure of Ceraeochrysa claveri (Navás) midgut endocrine cells in larva, pupa, and adult, and evaluated the side effects of ingested neem oil, a botanical insecticide obtained from the seeds of the neem tree (Azadirachta indica), on these cells. During the larval period, C. claveri were fed (ad libitum) Diatraea saccharalis (F.) eggs treated with neem oil at concentrations of 0.5%, 1%, or 2%. Transmission electron microscopy showed that two subtypes of endocrine cells, namely granular and vesicular, occurred in the midgut epithelium during the three stages of the life cycle. Both cell types did not reach the midgut lumen and were positioned basally in the epithelium. The endocrine cells did not show extensive infoldings of the basal plasma membrane, and there were numerous secretory granules in the basal region of the cytoplasm. In the granular endocrine cells, the granules were completely filled with a dense matrix. In the vesicular endocrine cells, the main secretory products consisted of haloed vesicles. Ultrastructural examination indicated that only the granular endocrine cells exhibited signs of morphologic changes of cell injury present in all life cycle stages after the larvae were chronically exposed to neem oil by ingestion. The major cellular damage consisted of dilatation and vesiculation of the rough endoplasmic reticulum and the development of smooth endoplasmic reticulum and mitochondrial swelling. Our data suggest that cytotoxic effects on midgut endocrine cells can contribute to a generalized disruption of the physiological processes in this organ due to a general alteration of endocrine function.

  8. Midgut volvulus due to jejunal diverticula: A case report

    Institute of Scientific and Technical Information of China (English)

    Jia-Li Hu; Wei-Zhong Chen

    2012-01-01

    Jejunal diverticulosis is uncommon and often asymptomatic.It can produce significant complications,and some complications are potentially life threatening and require early surgical treatment,such as obstruction,hemorrhage and perforation.There is no consensus on the management of this disease.Only a few cases of jejunal diverticulosis with midgut volvulus have been reported.We herein report a case of 57-year-old woman with jejunal diverticulosis causing small bowel volvulus who complained of intermittent upper abdominal pin-prick for 5 years that eventually progressed to a complete obstruction.The computed tomography scans revealed a mesenteric vessel "whirlpool" and laparotomy showed midgut volvulus secondary to jejunal diverticula.This case highlights jejunal diverticulosis causing small bowel volvulus as an uncommon mechanism of small bowel obstruction,which should be included in the differential diagnosis of small bowel obstruction.

  9. Heme crystallization in the midgut of triatomine insects.

    Science.gov (United States)

    Oliveira, Marcus F; Gandara, Ana Caroline P; Braga, Cláudia M S; Silva, José R; Mury, Flavia B; Dansa-Petretski, Marílvia; Menezes, Diego; Vannier-Santos, Marcos A; Oliveira, Pedro L

    2007-01-01

    Hemozoin (Hz) is a heme crystal produced by several blood-feeding organisms in order to detoxify free heme released upon hemoglobin (Hb) digestion. Here we show that heme crystallization also occurs in three species of triatomine insects. Ultraviolet-visible and infrared light absorption spectra of insoluble pigments isolated from the midgut of three triatomine species Triatoma infestans, Dipetalogaster maximus and Panstrongylus megistus indicated that all produce Hz. Morphological analysis of T. infestans and D. maximus midguts revealed the close association of Hz crystals to perimicrovillar membranes and also as multicrystalline assemblies, forming nearly spherical structures. Heme crystallization was promoted by isolated perimicrovillar membranes from all three species of triatomine bugs in vitro in heat-sensitive reactions. In conclusion, the data presented here indicate that Hz formation is an ancestral adaptation of Triatominae to a blood-sucking habit and that the presence of perimicrovillar membranes plays a central role in this process.

  10. Soil microflora over the closed Donbas mines

    Energy Technology Data Exchange (ETDEWEB)

    Kurdish, I.K.; Kisten, A.G.; Roi, A.A.; Iakovenko, E.A.; Bezborodov, V.A.

    2005-05-15

    Composition of soil air and microflora in different soil horizons (to 30 cm depth) over some closed Donbas coal mines has been studied. The intensive methane leakages have been found in some soil zones over the closed coal mines. This results in formation of explosive and inflammable gas mixtures. Methane content in soil gas mixtures could vary from trace amounts to 34 %. The essential decrease of oxygen (to 3-4%) and increasing (up to 5 %) of CO2 concentrations that evidences for active microbiological processes in such soils is marked in the latter case. Methanotrophic bacteria involving methane into biogenic cycle are widespread (up to 106 cells per 1 g) in methane-containing soils over the closed coal mines. Microorganisms of different physiological groups in the community with methanotrophic bacteria were found in the given technogenic econiche.

  11. [Soil microflora over the closed Donbas mines].

    Science.gov (United States)

    Kurdish, I K; Kisten', A G; Roĭ, A A; Iakovenko, E A; Bezborodov, V A

    2005-01-01

    Composition of soil air and microflora in different soil horizons (to 30 cm depth) over some closed Donbas coal mines has been studied. The intensive methane leakages have been found in some soil zones over the closed coal mines. This results in formation of explosive and inflammable gas mixtures. Methane content in soil gas mixtures could vary from trace amounts to 34 %. The essential decrease of oxygen (to 3-4%) and increasing (up to 5 %) of CO2 concentrations that evidences for active microbiological processes in such soils is marked in the latter case. Methanotrophic bacteria involving methane into biogenic cycle are widespread (up to 106 cells per 1 g) in methane-containing soils over the closed coal mines. Microorganisms of different physiological groups in the community with methanotrophic bacteria were found in the given technogenic econiche.

  12. Targeting intestinal microflora in inflammatory bowel disease

    Institute of Scientific and Technical Information of China (English)

    Mario Guslandi

    2006-01-01

    @@ TO THE EDITOR In their recent review article[1], Andoh and Fujiyama examined the various therapeutic approaches targeting intestinal microflora in patients with inflammatory bowel disease (IBD). I would like to provide some additional data to complete and update their comments. First of all, when considering the role of probiotics in 1BD treatment it must be emphasized that, in addition to Bifidobacteria, the Nissle 1917 E. coli strain and cocktails of microorganisms such as VSL # 3 mentioned in the article, other probiotic agents have been tested in the short- and long-term treatment of either ulcerative colitis and Crohn's disease, the results of those studies being reported in major international scientific journals.

  13. Fermentative hydrogen production by diverse microflora

    Energy Technology Data Exchange (ETDEWEB)

    Baghchehsaraee, B.; Nakhla, G.; Karamanev, D.; Margaritis, A. [Western Ontario Univ., London, ON (Canada). Dept. of Chemical and Biochemical Engineering

    2009-07-01

    This paper presented the results of a study in which hydrogen was produced from activated sludge. This diverse bacterial source has been compared to microflora from anaerobic digester sludge. Batch experiments were conducted at mesophilic (37 degrees C) and thermophilic (55 degrees C) temperatures. The hydrogen production yields with activated sludge at mesophilic and thermophilic temperatures were 0.25 and 0.93 mol H{sub 2}/mol glucose, respectively. The maximum hydrogen production rates with activated sludge in both temperatures were 4.2 mL/h. Anaerobic digester sludge showed higher hydrogen production yields and rates at both mesophilic and thermophilic temperatures. Repeated batch experiments with activated sludge resulted in increased hydrogen production in consecutive batches. However, the formation of lactic acid and ethanol resulted in unstable hydrogen production in the repeated batches.

  14. The fine structure of the midgut epithelium in Xerobiotus pseudohufelandi (Iharos, 1966 (Tardigrada, Eutardigrada, Macrobiotidae

    Directory of Open Access Journals (Sweden)

    Magdalena M. Rost-Roszkowska

    2013-04-01

    Full Text Available The aims of our studies were to describe the ultrastructure of the midgut epithelial cells of the eutardigrade Xerobiotus pseudohufelandi and to determine if there are any differences in the ultrastructure of midgut epithelial cells between males and females. The analysis was performed with the use of the light and transmission electron microscopes. In X. pseudohufelandi the midgut epithelium is composed of digestive cells, but in the anterior portion of the midgut a group of cells with different ultrastructure has been observed. Histochemical staining showed the accumulation of reserve material in the cytoplasm of digestive cells. We suggest that some of them fulfil the role of regenerative cells (crescent-like cells, midgut stem cells, whereas others are differentiating cells which form new digestive cells. No differences in the ultrastructure of the midgut epithelium between males and females were distinguished except in the amount of multivesicular bodies.

  15. Investigation of the midgut structure and ultrastructure in Cimex lectularius and Cimex pipistrelli (Hemiptera: Cimicidae)

    OpenAIRE

    Rost-Roszkowska, M. M.; Vilimova, J; Włodarczyk, A; Sonakowska, L; Kamińska, K; Kaszuba, F; Marchewka, A; Sadílek, D

    2016-01-01

    Cimicidae are temporary ectoparasites, which means that they cannot obtain food continuously. Both Cimex species examined here, Cimex lectularius (Linnaeus 1758) and Cimex pipistrelli (Jenyns 1839), can feed on a non-natal host, C. lectularius from humans on bats, C. pipistrelli on humans, but never naturally. The midgut of C. lectularius and C. pipistrelli is composed of three distinct regions—the anterior midgut (AMG), which has a sack-like shape, the long tube-shaped middle midgut (MMG), a...

  16. The oral microflora in obesity and type-2 diabetes

    OpenAIRE

    Shillitoe, Edward; Weinstock, Ruth; Kim, Taewan; Simon, Howard; Jessica, Planer; Noonan, Susan; Cooney, Robert

    2012-01-01

    Background: Type 2 diabetes mellitus (T2DM) is prevalent in people with obesity. It has been proposed that these conditions are related to specific features of the microflora of the mouth and lower gastrointestinal (GI) tract. Hyperglycemia often resolves quickly after Roux-en-Y gastric bypass (RYGB) but the role of the GI microflora cannot be examined easily because of reduced intestinal mobility. We propose that the study of microorganisms present in the mouth of patients undergoing RYGB wi...

  17. Fermentative hydrogen production from xylose using anaerobic mixed microflora

    Energy Technology Data Exchange (ETDEWEB)

    Lin, Chiu-Yue; Cheng, Chao-Hui [BioHydrogen Laboratory, Department of Hydraulic Engineering, Feng Chia University, P.O. Box 25-123, Taichung 40724 (Taiwan)

    2006-06-15

    Sewage sludge microflora were anaerobically cultivated in a chemostat-type anaerobic bioreactor at a temperature of 35+/-1{sup o}C, pH of 7.1 and hydraulic retention time of 12h to determine the hydrogen production efficiency from xylose (20g-COD/L). This enriched microflora was used as a seed in batch experiments to investigate the pH and substrate concentration effects on hydrogen-producing bioactivity. It is demonstrated that the enriched mesophilic sewage sludge microflora with a continuous feeding can produce hydrogen from xylose with hydrogen content of 32% (v/v) in the biogas. Each mole of xylose yields 0.7moles of hydrogen and each gram of biomass produces 0.038moles of hydrogen per day. According to the batch test results, changes in pH and xylose concentration could enhance the microflora hydrogen production activity. Batch cultivation of this mixed microflora at pH values of 6-7 and xylose concentrations of 20g-COD/L resulted in high hydrogen production with a yield of 1.92-2.25mol-H{sub 2}/mol-xylose. This value is comparable to that from an enrichment culture. Strategies based on pH and xylose concentration controls for optimal hydrogen production from xylose using sewage sludge microflora are proposed. (author)

  18. The Termite Gut Microflora as an Oxygen Sink: Microelectrode Determination of Oxygen and pH Gradients in Guts of Lower and Higher Termites.

    Science.gov (United States)

    Brune, A; Emerson, D; Breznak, J A

    1995-07-01

    Clark-type oxygen microelectrodes and glass pH microelectrodes, each with a tip diameter of Microcerotermes parvus (Haviland) revealed that there were extreme shifts as we moved posteriorly from the midgut proper (pH (apprx=)7) to the P1 segment of the hindgut (pH >10) and then to the P3 segment (paunch; pH (apprx=)7). The latter transition occurred at the short enteric valve (P2 segment) and within a distance of less than 500 (mu)m. In contrast, R. flavipes, which lacks a readily distinguishable P1 segment, did not possess a markedly alkaline region, and the pH around the midgut-hindgut junction was circumneutral. The oxic status of the peripheral hindgut lumen and its substantial oxygen consumption, together with previous reports of large numbers of aerobic and facultatively anaerobic bacteria in the hindgut microflora, challenge the notion that termite hindguts are a purely anoxic environment and, together with the steep axial pH gradients in higher termites, refine our concept of this tiny microbial habitat.

  19. Functional genomics of mountain pine beetle (Dendroctonus ponderosae midguts and fat bodies

    Directory of Open Access Journals (Sweden)

    Bearfield Jeremy C

    2010-03-01

    Full Text Available Abstract Background The mountain pine beetle (Dendroctonus ponderosae is a significant coniferous forest pest in western North America. It relies on aggregation pheromones to colonize hosts. Its three major pheromone components, trans-verbenol, exo-brevicomin, and frontalin, are thought to arise via different metabolic pathways, but the enzymes involved have not been identified or characterized. We produced ESTs from male and female midguts and associated fat bodies and used custom oligonucleotide microarrays to study gene expression patterns and thereby made preliminary identification of pheromone-biosynthetic genes. Results Clones from two un-normalized cDNA libraries were directionally sequenced from the 5' end to yield 11,775 ESTs following sequence cleansing. The average read length was 550 nt. The ESTs clustered into 1,201 contigs and 2,833 singlets (4,034 tentative unique genes. The ESTs are broadly distributed among GO functional groups, suggesting they reflect a broad spectrum of the transcriptome. Among the most represented genes are representatives of sugar-digesting enzymes and members of an apparently Scolytid-specific gene family of unknown function. Custom NimbleGen 4-plex arrays representing the 4,034 tentative unique genes were queried with RNA from eleven different biological states representing larvae, pupae, and midguts and associated fat bodies of unfed or fed adults. Quantitative (Real-Time RT-PCR (qRT-PCR experiments confirmed that the microarray data accurately reflect expression levels in the different samples. Candidate genes encoding enzymes involved in terminal steps of biosynthetic pathways for exo-brevicomin and frontalin were tentatively identified. Conclusions These EST and microarray data are the first publicly-available functional genomics resources for this devastating forestry pest.

  20. The RNA interference pathway affects midgut infection- and escape barriers for Sindbis virus in Aedes aegypti

    Directory of Open Access Journals (Sweden)

    Olson Ken E

    2010-04-01

    Full Text Available Abstract Background The RNA interference (RNAi pathway acts as an innate antiviral immune response in Aedes aegypti, modulating arbovirus infection of mosquitoes. Sindbis virus (SINV; family: Togaviridae, genus: Alphavirus is an arbovirus that infects Ae. aegypti in the laboratory. SINV strain TR339 encounters a midgut escape barrier (MEB during infection of Ae. aegypti. The nature of this barrier is not well understood. To investigate the role of the midgut as the central organ determining vector competence for arboviruses, we generated transgenic mosquitoes in which the RNAi pathway was impaired in midgut tissue of bloodfed females. We used these mosquitoes to reveal effects of RNAi impairment in the midgut on SINV replication, midgut infection and dissemination efficiencies, and mosquito longevity. Results As a novel tool for studying arbovirus-mosquito interactions, we engineered a transgenic mosquito line with an impaired RNAi pathway in the midgut of bloodfed females by silencing expression of the Aa-dcr2 gene. In midgut tissue of the transgenic Carb/dcr16 line, Aa-dcr2 expression was reduced ~50% between 1-7 days post-bloodmeal (pbm when compared to the recipient mosquito strain. After infection with SINV-TR339EGFP, Aa-dcr2 expression levels were enhanced in both mosquito strains. In the RNAi pathway impaired mosquito strain SINV titers and midgut infection rates were significantly higher at 7 days pbm. There was also a strong tendency for increased virus dissemination rates among the transgenic mosquitoes. Between 7-14 days pbm, SINV was diminished in midgut tissue of the transgenic mosquitoes. Transgenic impairment of the RNAi pathway and/or SINV infection did not affect longevity of the mosquitoes. Conclusions We showed that RNAi impaired transgenic mosquitoes are a useful tool for studying arbovirus-mosquito interactions at the molecular level. Following ingestion by Ae. aegypti, the recombinant SINV-TR339EGFP was confronted with both

  1. Fermentative hydrogen production by diverse microflora

    Energy Technology Data Exchange (ETDEWEB)

    Baghchehsaraee, B.; Nakhla, G.; Karamanev, D.; Margaritis, A. [Dept. of Chemical and Biochemical Engineering, Univ. of Western Ontario, London, Ontario (Canada)

    2009-07-01

    'Full text': In this study of hydrogen production with activated sludge, a diverse bacterial source has been investigated and compared to microflora from anaerobic digester sludge, which is less diverse. Batch experiments were conducted at mesophilic (37 {sup o}C) and thermophilic (55 {sup o}C) temperatures. The hydrogen production yields with activated sludge at 37 {sup o}C and 55 {sup o}C were 0.25 and 0.93 mol H{sub 2}/mol glucose, respectively. The maximum hydrogen production rates with activated sludge in both temperatures were 4.2 mL/h. Anaerobic digester sludge showed higher hydrogen production yields and rates at both mesophilic and thermophilic temperatures. The results of repeated batch experiments with activated sludge showed an increase in the hydrogen production during the consecutive batches. However, hydrogen production was not stable along the repeated batches. The observed instability was due to the formation of lactic acid and ethanol. (author)

  2. Treponema denticola in microflora of bovine periodontitis

    Directory of Open Access Journals (Sweden)

    Ana Carolina Borsanelli

    2015-03-01

    Full Text Available Periodontitis in cattle is an infectious purulent progressive disease associated with strict anaerobic subgingival biofilm and is epidemiologically related to soil management at several locations of Brazil. This study aimed to detect Treponema species in periodontal pockets of cattle with lesions deeper than 5mm in the gingival sulcus of 6 to 24-month-old animals considered periodontally healthy. We used paper cones to collect the materials, after removal of supragingival plaques, and kept frozen (at -80°C up to DNA extraction and polymerase chain reaction (PCR using T. amylovorum, T. denticola, T. maltophilum, T. medium and T. vincentii primers. In periodontal pocket, it was possible to identify by PCR directly, the presence of Treponema amylovorum in 73% of animals (19/26, T. denticola in 42.3% (11/26 and T. maltophilum in 54% (14/26. Among the 25 healthy sites, it was possible to identify T. amylovorum in 18 (72%, T. denticola in two (8% and T. maltophilum in eight (32%. Treponema medium and T. vincentii were not detected over all 51 evaluated samples. The presence of Treponema amylovorum, T. maltophilum and, in particular, the widely recognized T. denticola in subgingival microflora brings an original and potencially important contribution in studies of the bovine periodontitis.

  3. Transcriptional Signatures in Response to Wheat Germ Agglutinin and Starvation in Drosophila melanogaster Larval Midgut

    Science.gov (United States)

    One function of plant lectins such as wheat germ agglutinin (WGA) is to serve as defenses against herbivorous insects. The midgut is one critical site affected by dietary lectins. We observed marked cellular, structural, and gene expression changes in the midguts of Drosophila melanogaster third-i...

  4. Midgut pain due to an intussuscepting terminal ileal lipoma: a case report.

    LENUS (Irish Health Repository)

    Abbasakoor, Noormuhammad O

    2010-01-01

    ABSTRACT: INTRODUCTION: The occurrence of intussusception in adults is rare. The condition is found in 1 in 1300 abdominal operations and 1 in 100 patients operated for intestinal obstruction. The child to adult ratio is 20:1. CASE PRESENTATION: A 52-year-old Irish Caucasian woman was investigated for a 3-month history of intermittent episodes of colicky midgut pain and associated constipation. Ileocolonoscopy revealed a pedunculated lesion in the terminal ileum prolapsing into the caecum. Computed tomography confirmed a smooth-walled, nonobstructing, low density intramural lesion in the terminal ileum with secondary intussusception. A laparoscopic small bowel resection was performed. Histology revealed a large pedunculated polypoidal mass measuring 4 x 2.5 x 2 cm consistent with a submucosal lipoma. She had complete resolution of her symptoms and remained well at 12-month follow-up. CONCLUSION: This case highlights an unusual cause of incomplete small bowel obstruction successfully treated through interdisciplinary cooperation. Ileal lipomas are not typically amenable to endoscopic removal and require resection. This can be successfully achieved via a laparoscopic approach with early restoration of premorbid functioning.

  5. Midgut pain due to an intussuscepting terminal ileal lipoma: a case report

    Directory of Open Access Journals (Sweden)

    Ryan Barbara

    2010-02-01

    Full Text Available Abstract Introduction The occurrence of intussusception in adults is rare. The condition is found in 1 in 1300 abdominal operations and 1 in 100 patients operated for intestinal obstruction. The child to adult ratio is 20:1. Case presentation A 52-year-old Irish Caucasian woman was investigated for a 3-month history of intermittent episodes of colicky midgut pain and associated constipation. Ileocolonoscopy revealed a pedunculated lesion in the terminal ileum prolapsing into the caecum. Computed tomography confirmed a smooth-walled, nonobstructing, low density intramural lesion in the terminal ileum with secondary intussusception. A laparoscopic small bowel resection was performed. Histology revealed a large pedunculated polypoidal mass measuring 4 × 2.5 × 2 cm consistent with a submucosal lipoma. She had complete resolution of her symptoms and remained well at 12-month follow-up. Conclusion This case highlights an unusual cause of incomplete small bowel obstruction successfully treated through interdisciplinary cooperation. Ileal lipomas are not typically amenable to endoscopic removal and require resection. This can be successfully achieved via a laparoscopic approach with early restoration of premorbid functioning.

  6. Morpho-functional characterization and esterase patterns of the midgut of Tribolium castaneum Herbst, 1797 (Coleoptera: Tenebrionidae) parasitized by Gregarina cuneata (Apicomplexa: Eugregarinidae).

    Science.gov (United States)

    Gigliolli, Adriana A Sinópolis; Lapenta, Ana Silva; Ruvolo-Takasusuki, Maria Claudia Colla; Abrahão, Josielle; Conte, Hélio

    2015-09-01

    Tribolium castaneum (Coleoptera: Tenebrionidae) is a common pest of stored grains and byproducts and is normally infected by Gregarina cuneata (Apicomplexa: Eugregarinidae). The life cycle of this parasite includes the sporozoite, trophozoite, gamont, gametocyte, and oocyst stages, which occur between the epithelium and lumen of the host's midgut. This study aims to describe the morphofunctional alterations in the midgut and determine the esterase patterns in T. castaneum when parasitized by gregarines. To achieve this purpose, midguts of adult insects were isolated, processed, and analysed using light and electron microscopy. We determined total protein content, amylase activity, and the expression and related activities of the esterases by using polyacrylamide gel electrophoresis (PAGE). The midgut of T. castaneum is formed by digestive, regenerative, and endocrine cells. The effects of parasitism on the digestive cells are severe, because the gregarines remain attached to these cells to absorb all the nutrients they need throughout their development. In these cells, the most common alterations observed include expansion and fragmentation of the rough endoplasmic reticulum, development of the smooth endoplasmic reticulum, changes in mitochondrial cristae, cytoplasmic vacuolization, formation of myelin structures, spherites, large intercellular spaces, autophagic vesicles, expansion of the basal labyrinth, and cytoplasmic protrusions. Deposits of glycogen granules were also observed. Amylase activity was reduced in parasitized insects. Regenerative cells were found in disorganized crypts and did not differentiate into new cells, thus, compromising the restoration of the damaged epithelium. Though few morphological alterations were observed in the endocrine cells, results suggest that the synthesis and/or release of hormones might be impaired. Nine esterases (EST-1 to 9) were identified in the midgut of T. castaneum and were expressed in varying levels in response

  7. The vaginal microflora in relation to gingivitis

    Directory of Open Access Journals (Sweden)

    Weibel Marianne

    2009-01-01

    Full Text Available Abstract Background Gingivitis has been linked to adverse pregnancy outcome (APO. Bacterial vaginosis (BV has been associated with APO. We assessed if bacterial counts in BV is associated with gingivitis suggesting a systemic infectious susceptibilty. Methods Vaginal samples were collected from 180 women (mean age 29.4 years, SD ± 6.8, range: 18 to 46, and at least six months after delivery, and assessed by semi-quantitative DNA-DNA checkerboard hybridization assay (74 bacterial species. BV was defined by Gram stain (Nugent criteria. Gingivitis was defined as bleeding on probing at ≥ 20% of tooth sites. Results A Nugent score of 0–3 (normal vaginal microflora was found in 83 women (46.1%, and a score of > 7 (BV in 49 women (27.2%. Gingivitis was diagnosed in 114 women (63.3%. Women with a diagnosis of BV were more likely to have gingivitis (p = 0.01. Independent of gingival conditions, vaginal bacterial counts were higher (p Prevotella bivia (p Prevotella disiens (p P. bivia, P. disiens, M. curtisii and M. mulieris (all at the p 1.0 × 104 cells and a diagnosis of gingivitis was 3.9 for P. bivia (95% CI 1.5–5.7, p P. disiens (95%CI: 1.8–7.5, p P. bivia (odds ratio: 5.3, 95%CI: 2.6 to 10.4, p P. disiens (odds ratio: 4.4, 95% CI: 2.2 to 8.8, p Conclusion Higher vaginal bacterial counts can be found in women with BV and gingivitis in comparison to women with BV but not gingivitis. P. bivia and P. disiens may be of specific significance in a relationship between vaginal and gingival infections.

  8. New Horizons for the Infectious Diseases Specialist: How Gut Microflora Promote Health and Disease

    OpenAIRE

    Rabizadeh, Shervin; Sears, Cynthia

    2008-01-01

    The human intestine provides an expansive interface for interactions with the microflora. Increasing data support the hypothesis that host–microflora relationships are markedly dynamic, contributing to host health and disease pathogenesis. Despite outnumbering human cells 10-fold, the microflora most often assist the host through symbiotic relationships. The microflora are involved in maximizing host utilization of nutrients, induction of host immune responses, and promotion of intestinal cel...

  9. INTESTINAL VIROME AND NORMAL MICROFLORA OF HUMAN: FEATURES OF INTERACTION

    Directory of Open Access Journals (Sweden)

    Bobyr V.V.

    2015-05-01

    Full Text Available Summary: Intestinal bacteria defend the host organism and narrow pathogenic bacterial colonization. However, the microbiome effect to enteric viruses is unexplored largely as well as role of microbiota in the pathogenesis of viral infections in general. This review focuses on precisely these issues. Keywords: microbiome, virome, normal microflora, enteric viruses, contagiousness. In this review article, facts about viral persistence in the human gut are summarized. It is described the role of viral populations during health and diseases. After analyzing of the literary facts it was concluded that the gastrointestinal tract is an environment for one from the most complex microbial ecosystems, which requires of more deeper study of its composition, role in physiological processes, as well as the dynamics of changes under influence of the environment. Normal microflora performs a different important functions providing the physiological homeostasis of the human body, including, in particular, an important role in the human metabolic processes, supporting of homeostasis, limiting of colonization by infectious bacteria. The multifactorial significance of the normal gastrointestinal microflora can be divided into immunological, structural and metabolic functions. At the same time, interaction between intestinal microflora and enteric viruses has not been studied largely. In recent years, much attention is paid to study of viruses-bacteria associations, and it is possible, obtained results should change our understanding of microbiota role in the systematic pathogenesis of the diseases with viral etiology. In contrast to the well-known benefits of normal microflora to the host, the viruses can use intestinal microflora as a trigger for replication at the optimal region. Recent studies give a reason for assumption that depletion of normal microflora with antibiotics can determining the antiviral effect. Thus, the role of commensal bacteria in viral

  10. Altered protoxin activation by midgut enzymes from a Bacillus thuringiensis resistant strain of Plodia interpunctella.

    Science.gov (United States)

    Oppert, B; Kramer, K J; Johnson, D E; MacIntosh, S C; McGaughey, W H

    1994-02-15

    Processing of Bacillus thuringiensis protoxins to toxins by midgut proteinases from a strain of the Indianmeal moth, Plodia interpunctella (Hubner), resistant to B. thuringiensis subspecies entomocidus (HD-198) was slower than that by midgut proteinases from the susceptible parent strain or a strain resistant to B. thuringiensis subspecies kurstaki (HD-1, Dipel). Midgut extracts from entomocidus-resistant insects exhibited five-fold lower activity toward the synthetic substrate alpha-N-benzoyl-DL-arginine rho-nitroanilide than extracts from susceptible or kurstaki-resistant insects. Midgut enzymes from susceptible or kurstaki-resistant insects converted the 133 kDa CryIA(c) protoxin to 61-63 kDa proteins, while incubations with entomocidus-resistant enzymes resulted in predominantly products of intermediate size, even with increased amounts of midgut extract. The 61-63 kDa proteins were only produced by entomocidus-resistant midgut extracts after long term incubations with the protoxin. The data suggest that altered protoxin activation by midgut proteinases is involved in some types of insect resistance to B. thuringiensis.

  11. Ultrastructural changes of the midgut epithelium in Isohypsibius granulifer granulifer Thulin, 1928 (Tardigrada: Eutardigrada) during oogenesis.

    Science.gov (United States)

    Rost-Roszkowska, Magdalena M; Poprawa, Izabela; Wójtowicz, Maria; Kaczmarek, Lukasz

    2011-04-01

    The midgut epithelium of Isohypsibius granulifer granulifer (Eutardigrada) is composed of columnar digestive cells. At its anterior end, a group of cells with cytoplasm which differs from the cytoplasm of digestive cells is present. Probably, those cells respond to crescent-like cells (midgut regenerative cells) described for some tardigrade species. Their mitotic divisions have not been observed. We analyzed the ultrastructure of midgut digestive cells in relation to five different stages of oogenesis (previtellogenesis, beginning of the vitellogenesis, vitellogenesis--early choriogenesis, vitellogenesis--middle choriogenesis, late choriogenesis). In the midgut epithelium cells, the gradual accumulation of glycogen granules, lipid droplets and structures of varying electron density occurs. During vitellogenesis and choriogenesis, in the cytoplasm of midgut cells we observed the increasing number of organelles which are responsible for the intensive synthesis of lipids, proteins and saccharides such as cisterns of endoplasmic reticulum and Golgi complexes. At the end of oogenesis, autophagy also intensifies in midgut epithelial cells, which is probably caused by the great amount of reserve material. Midgut epithelium of analyzed species takes part in the yolk precursor synthesis.

  12. Resistance to Bacillus thuringiensis by the Indian meal moth, Plodia interpunctella: comparison of midgut proteinases from susceptible and resistant larvae.

    Science.gov (United States)

    Johnson, D E; Brookhart, G L; Kramer, K J; Barnett, B D; McGaughey, W H

    1990-03-01

    Midgut homogenates from susceptible and resistant strains of the Indian meal moth, Plodia interpunctella, were compared for their ability to activate the entomocidal parasporal crystal protein from Bacillus thuringiensis. The properties of midgut proteinases from both types of larvae were also examined. Electrophoretic patterns of crystal protein from B. thuringiensis subspecies kurstaki (HD-1) and aizawai (HD-133 and HD-144) were virtually unchanged following digestion by either type of midgut homogenate. Changes in pH (9.5 to 11.5) or midgut homogenate concentration during digestion failed to substantially alter protein electrophoretic patterns of B. thuringiensis HD-1 crystal toxin. In vitro toxicity of crystal protein activated by either type of midgut preparation was equal toward cultured insect cells from either Manduca sexta or Choristoneura fumiferana. Electrophoresis of midgut extracts in polyacrylamide gels containing gelatin as substrate also yielded matching mobility patterns of proteinases from both types of midguts. Quantitation of midgut proteolytic activity using tritiated casein as a substrate revealed variation between midgut preparations, but no statistically significant differences between proteolytic activities from susceptible and resistant Indian meal moth larvae. Inhibition studies indicated that a trypsin-like proteinase with maximal activity at pH 10 is a major constituent of Indian meal moth midguts. The results demonstrated that midguts from susceptible and resistant strains of P. interpunctella are similar both in their ability to activate B. thuringiensis protoxin and in their proteolytic activity.

  13. MIKROFLORA PADA TEMPOYAK The Microflora of Tempoyak

    Directory of Open Access Journals (Sweden)

    Hasanuddin Hasanuddin

    2012-05-01

    Full Text Available Tempoyak is a traditional Bengkulu fermented food prepared from spontaneous fermentation of durian (Durio zibethi- nus by wild microorganisms. Tempoyak is found not only in Bengkulu but also as long as Sumatera island with differ- ent names, and also in Malaysia. Tempoyak is a Melayu term meaning fermented durian. The research was conducted to identify microflora in tempoyak, and to select the species of microorganisms which were able to ferment durian ef- fectivelly. The data in this study were laboratory analysis. Samples were collected weekly in a month from productions centre of tempoyak and traditional markets, and analyzed microbiologically to determine and identify the bacteria, yeasts and molds in tempoyak. There were four specieses of lactic acid bacteria found in fermented durian namely Pediococcus acidilactici, Lactobacillus plantarum Lactobacillus curvatus and Leuconostoc mesentroides. While two specieses which were not lactic acid bacteria namely Staphylococcus saprophyticus and Micrococcus varians. One species of yeast was Kluyveromyces marxianus. The molds were identified as Rhizopus oryzae, Monilia sitophila, Mucor roxii, Aspergillus repens and Penicillium sp. Among these specieses three specieses were produced lactic acid namelly Rhizopus oryzae, Monilia sitophila,and Mucor roxii. While Aspergillus ripens and Penicillium sp did not pro- duce lactic acid so these specieses might not function in fermentation process. ABSTRAK Tempoyak adalah makanan khas Bengkulu, yang dibuat dari fermentasi spontan daging buah durian (Durio zibet- hinus oleh mikroorganisme liar. Tempoyak juga bisa dijumpai di berbagai daerah Indonesia di sepanjang pulau Sumatera dengan nama yang berbeda, bahkan sampai ke Malaysia. Istilah tempoyak berasal dari terminolgi Melayu yang artinya adalah durian fermentasi. Penelitian ini bertujuan untuk mengidentifikasi mikroorganisme yang terdapat pada tempoyak. Data dari penelitian ini adalah data primer dari analisa

  14. Characterization of a midgut-specific chitin synthase gene (LmCHS2) responsible for biosynthesis of chitin of peritrophic matrix in Locusta migratoria.

    Science.gov (United States)

    Liu, Xiaojian; Zhang, Huanhuan; Li, Sheng; Zhu, Kun Yan; Ma, Enbo; Zhang, Jianzhen

    2012-12-01

    Chitin, an essential component of peritrophic matrix (PM), is produced by a series of biochemical reactions. Chitin synthase plays a crucial role in chitin polymerization in chitin biosynthetic pathway. In this study, we identified and characterized a full-length cDNA of chitin synthase 2 gene (LmCHS2) from Locusta migratoria. The cDNA contains an open reading frame of 4569 nucleotides that encode 1523 amino acid residues, and 76- and 373-nucleotides for 5'- and 3'-noncoding regions, respectively. Analysis of LmCHS2 transcript in different tissues of the locust by using real-time quantitative PCR indicated that LmCHS2 was exclusively expressed in midgut and gastric caeca (a part of the midgut). The highest expression was found in the anterior midgut with a decline of the transcript level from the anterior to posterior regions. During growth and development of locusts, there was only a slight expression in eggs, but the expression gradually increased from nymphs to adults. In situ hybridization further revealed that LmCHS2 transcript mainly presented in the apical regions of brush border forming columnar cells of gastric caeca. LmCHS2 dsRNA was injected to fifth-instar nymphs to further explore biological functions of LmCHS2. Significantly down-regulated transcript of LmCHS2 resulted in a cessation of feeding and a high mortality of the insect. However, no visible abnormal morphological change of locusts was observed until insects molted to adults. After dissection, we found that the average length of midguts from the LmCHS2 dsRNA-injected locusts was shorter than that of the control insects that were injected with dsGFP. Furthermore, microsection of midguts showed that the PM of the LmCHS2 dsRNA-injected nymphs was amorphous and thin as compared with the controls. Our results demonstrate that LmCHS2 is responsible for the biosynthesis of chitin associated with PM and plays an essential role in locust growth and development.

  15. In silico modelling of the human intestinal microflora

    NARCIS (Netherlands)

    Kamerman, DJ; Wilkinson, MHF; Sloot, P; Tan, CJK; Dongarra, JJ; Hoekstra, AG

    2002-01-01

    The ecology of the human intestinal microflora and its interaction with the host are poorly understood. Though more and more data are being acquired, in part using modern molecular methods, development of a quantitative theory has not kept pace with this development. This is in part due to the compl

  16. In Silico Modelling of the Human Intestinal Microflora

    NARCIS (Netherlands)

    Kamerman, Derk Jan; Wilkinson, Michael H.F.

    2002-01-01

    The ecology of the human intestinal microflora and its interaction with the host are poorly understood. Though more and more data are being acquired, in part using modern molecular methods, development of a quantitative theory has not kept pace with this development. This is in part due to the compl

  17. Disease suppressive soilless culture systems; characterisation of its microflora

    NARCIS (Netherlands)

    Postma, J.; Willemsen-de Klein, M.J.E.I.M.; Rattink, H.; Os, van E.A.

    2001-01-01

    The trend in glasshouse horticulture has always been to start culture systems as aseptic as possible. However, several root diseases still cause problems under these conditions. The present paper shows the importance of the microflora to suppress Pythium aphanidermatum, a fungal root pathogen which

  18. Isolation and characterization of bacteria from midgut of the rice water weevil (Coleoptera: Curculionidae).

    Science.gov (United States)

    Lu, Fang; Kang, Xiaoying; Jiang, Cong; Lou, Binggan; Jiang, Mingxing; Way, Michael O

    2013-10-01

    Gut bacteria are known to play important and often essential roles in the biology of insects. Theoretically, they can be genetically manipulated, then reintroduced into insects to negatively modify specific biological features. The weevil superfamily Curculionoidea is one of the most species-rich and successful animal groups on earth, but currently the overall knowledge of the bacterial communities in weevils and their associations with hosts is still limited. In this study, we isolated and characterized the bacteria in the midgut of an invasive weevil, Lissorhoptrus oryzophilus Kuschel, by culturing methods. Female adults of this weevil were collected from four different geographic regions of the United States and mainland China. Sequencing of the bacterial 16S rRNA amplicons demonstrated that the major culturable gut bacteria of rice water weevil are γ-proteobacteria and Bacilli. The gut bacterial composition differs among regions, with many of the bacteria isolated from only a single region while several were detected from more than one region. Overall, the diversity of gut bacteria in rice water weevil is relatively low. The possible origins of certain bacteria are discussed in relation to the weevil, rice plant, and bacteria.

  19. The oral microflora--friend or foe? Can we decide?

    Science.gov (United States)

    Marsh, P D; Percival, R S

    2006-08-01

    This paper discusses the factors that determine whether the oral microflora play a beneficial or detrimental role in the health of an individual during their life. The resident microflora of the mouth is diverse, and distinct from that found elsewhere in the body due to its unique biological and physical properties. This natural microflora is essential for the normal development of the physiology of the host, and contributes to the host defences by excluding exogenous micro-organisms. The oral microflora varies in composition on distinct surfaces (e.g. teeth, mucosa), and at sites on a specific surface (e.g. fissures, gingival crevice), demonstrating that subtle properties of a habitat influence the ability of individual species to colonise and dominate. The composition of these oral microbial communities remains relatively stable over time (microbial homeostasis). This stability does not indicate a passive relationship with the host, but reflects a dynamic balance among the component species. However, this stability can be perturbed by significant changes to the oral environment or in a person's life-style that occur during the life of an individual. Alterations in diet, medication, smoking, saliva flow, denture wearing, general health, etc, can lead to overgrowth by previously minor components of the oral microflora, which can predispose a site to disease. Likewise, the immune response can wane in old age, which may result in colonisation by exogenous and often pathogenic micro-organisms. Oral micro-organisms can also act as opportunistic pathogens, and cause serious disease elsewhere in the body. Therefore, active oral health care management is needed in order to maintain microbial homeostasis throughout life to ensure that we reap the benefits of our resident oral bacteria and not suffer from their mischief.

  20. Intrauterine midgut volvulus without malrotation:Diagnosis from the 'coffee bean sign'

    Institute of Scientific and Technical Information of China (English)

    Jun Seok Park; Seong Jae Cha; Beom Gyu Kim; Yong Seok Kim; Yoo Shin Choi; In Talk Chang; Gwang Jun Kim; Woo Seok Lee; Gi Hyeon Kim

    2008-01-01

    Fetal midgut volvulus is quite rare,and most cases areassociated with abnormalities of intestinal rotation orfixation.We report a case of midgut volvulus withoutmalrotation,associated with a meconium pellet,duringthe gestation period.This 2.79 kg,33-wk infant was bornvia a spontaneous vaginal delivery caused by pretermlabor.Prenatal ultrasound showed dilated bowel loopswith the appearance of a 'coffee bean sign" This patienthad an unusual presentation with a distended abdomenshowing skin discoloration.An emergency laparotomyrevealed a midgut volvulus and a twisted small bowel,caused by complicated meconium ileus.Such nonspecificprenatal radiological signs and a low index of suspicionof a volvulus during gestation might delay appropriatesurgical management and result in ischemic necrosis of the bowel.Preterm labor,specific prenatal sonographicfindings (for example,the coffee bean sign) and bluishdiscoloration of the abdominal wall could suggestintrauterine midgut volvulus requiring prompt surgicalintervention.

  1. Identification of the midgut microbiota of An. stephensi and An. maculipennis for their application as a paratransgenic tool against malaria.

    Directory of Open Access Journals (Sweden)

    Navid Dinparast Djadid

    Full Text Available The midgut microbiota associated with Anopheles stephensi and Anopheles maculipennis (Diptera: Culicidae was investigated for development of a paratransgenesis-based approach to control malaria transmission in Eastern Mediterranean Region (EMR. Here, we present the results of a polymerase chain reaction (PCR and biochemical-based approaches to identify the female adult and larvae mosquitoe microbiota of these two major malaria vectors, originated from South Eastern and North of Iran. Plating the mosquito midgut contents from lab-reared and field-collected Anopheles spp. was used for microbiota isolation. The gram-negative and gram-positive bacterial colonies were identified by Gram staining and specific mediums. Selected colonies were identified by differential biochemical tests and 16S rRNA gene sequence analysis. A number of 10 An. stephensi and 32 An. maculipennis adult mosquitoes and 15 An. stephensi and 7 An. maculipennis larvae were analyzed and 13 sequences of 16S rRNA gene bacterial species were retrieved, that were categorized in 3 classes and 8 families. The majority of the identified bacteria were belonged to the γ-proteobacteria class, including Pseudomonas sp. and Aeromonas sp. and the others were some closely related to those found in other vector mosquitoes, including Pantoea, Acinetobacter, Brevundimonas, Bacillus, Sphingomonas, Lysinibacillus and Rahnella. The 16S rRNA sequences in the current study aligned with the reference strains available in GenBank were used for construction of the phylogenetic tree that revealed the relatedness among the bacteria identified. The presented data strongly encourage further investigations, to verify the potential role of the detected bacteria for the malaria control in Iran and neighboring countries.

  2. Effect of midgut proteolytic activity on susceptibility of lepidopteran larvae to Bacillus thuringiensis subsp. kurstaki

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    Reza eTalaei-Hassanloui

    2014-01-01

    Full Text Available Bacillus thuringiensis (Bt is the most effective microbial control agent for controlling numerous species from different insect orders. All subspecies and strains of B. thuringiensis can produce a spore and a crystalline parasporal body. This crystal which contains proteinaceous protoxins is dissolved in the alkaline midgut, the resulting molecule is then cleaved and activated by proteolytic enzymes and acts as a toxin. An interesting aspect of this activation process is that variations in midgut pH and protease activity have been shown to account for the spectrum of some Bt proteins activity. Thus, an important factor that could be a determinant of toxin activity is the presence of proteases in the midgut microenvironment of susceptible insects. Reciprocally, any alteration in the midgut protease composition of the host can result in resistance to Bt. Here in this paper, we reviewed this processes in general and presented our assays to reveal whether resistance mechanism to Bt in Diamondback Moth larvae could be due to the function of the midgut proteases? We estimated LC50 for both probable susceptible and resistant populations in laboratory and greenhouse tests. Then, the midgut protease activities of the B. thuringiensis ind

  3. Midgut barrier imparts selective resistance to filarial worm infection in Culex pipiens pipiens.

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    Michelle L Michalski

    Full Text Available Mosquitoes in the Culex pipiens complex thrive in temperate and tropical regions worldwide, and serve as efficient vectors of Bancroftian lymphatic filariasis (LF caused by Wuchereria bancrofti in Asia, Africa, the West Indies, South America, and Micronesia. However, members of this mosquito complex do not act as natural vectors for Brugian LF caused by Brugia malayi, or for the cat parasite B. pahangi, despite their presence in South Asia where these parasites are endemic. Previous work with the Iowa strain of Culex pipiens pipiens demonstrates that it is equally susceptible to W. bancrofti as is the natural Cx. p. pipiens vector in the Nile Delta, however it is refractory to infection with Brugia spp. Here we report that the infectivity barrier for Brugia spp. in Cx. p. pipiens is the mosquito midgut, which inflicts internal and lethal damage to ingested microfilariae. Following per os Brugia exposures, the prevalence of infection is significantly lower in Cx. p. pipiens compared to susceptible mosquito controls, and differs between parasite species with <50% and <5% of Cx. p. pipiens becoming infected with B. pahangi and B. malayi, respectively. When Brugia spp. mf were inoculated intrathoracically to bypass the midgut, larvae developed equally well as in controls, indicating that, beyond the midgut, Cx. p. pipiens is physiologically compatible with Brugia spp. Mf isolated from Cx. p. pipiens midguts exhibited compromised motility, and unlike mf derived from blood or isolated from the midguts of Ae. aegypti, failed to develop when inoculated intrathoracically into susceptible mosquitoes. Together these data strongly support the role of the midgut as the primary infection barrier for Brugia spp. in Cx. p. pipiens. Examination of parasites recovered from the Cx. p. pipiens midgut by vital staining, and those exsheathed with papain, suggest that the damage inflicted by the midgut is subcuticular and disrupts internal tissues. Microscopic studies

  4. Study of pulp microflora in patients with cardiovascular diseases

    Directory of Open Access Journals (Sweden)

    M.A. Safarov

    2010-06-01

    Full Text Available 335 patients aged 20 to 60 years with various parodontitis inflammatory diseases have been selected for research. All patients have been divided into four groups of different age: with rheumatism - 96 persons, with heart ischemic illness - 82 persons, with arterial hypertension - 89 persons, with neurocirculatory dystonia - 68 persons. The presented results of supervision show diagnostic significant changes of pulp microflora with odontogenic infection in patients, suffering cardiovascular diseases

  5. Impact of salinomycin on the intestinal microflora of broiler chickens

    Directory of Open Access Journals (Sweden)

    Pedersen Karl

    2007-10-01

    Full Text Available Abstract Background The ionophoric coccidiostat salinomycin is widely used in chicken feed. In the near future the use of ionophore coccidiostats may be banned as has been the case for other antimicrobial growth promoters. This study was conducted to examine the effect of salinomycin on Campylobacter jejuni infection and on the composition of the caecal microflora in broiler chickens. Methods An experimental infection study was carried out in isolators and the intestinal microflora was analyzed using quantitative cultivation, denaturant gradient gel electrophoresis (DGGE, cloning and sequencing. Results We found no effect of salinomycin on C. jejuni but salinomycin significantly affected the composition of the microflora. In addition, salinomycin significantly reduced the prevalence of Clostridium perfringens and we observed a significant increase (62% in the mean body weight of salinomycin treated chickens compared to un-treated controls. Conclusion Termination of the use of ionophore coccidiostats will not affect food safety related to campylobacter, but will increase the risk of necrotic enteritis in the broilers.

  6. The contribution of gut microflora to paracetamol metabolism

    Directory of Open Access Journals (Sweden)

    Bojić Gordana

    2014-01-01

    Full Text Available During the last decade, the important role of gut microflora as a special organ of the gastrointestinal system in the metabolism of drugs is well known. The aim of this study was to evaluate the role of the gut microecological population with enzymatic systems, especially beta-lyase, in the metabolism of paracetamol in mice. Two groups of 20 white male laboratory mice BALB/c, body weight 32+/-1.5 kg, were treated orally with neomycin sulphate (500 mg/kg in saline solution and saline solution (10 ml/kg twice daily for three days. After the treatment, the animals were given paracetamol dissolved in saline solution (200 mg/kg intraperitoneally. The total amount of excreted paracetamol in 8 hours’ collected urine was unchanged. A difference between treated and control mice was observed regarding a highly significant reduction in the excretion of 3-methylthiometabolites. A decrease in the excretion of thiomethyl metabolites was found in the control group compared to the experimental mice. Gut microflora had a great influence on the formation of metabolic precursors, thiomethyl-conjugates, and their oxidabile products. It is obvious that the ecosystem of gut microflora has an important role in the metabolism of paracetamol resulting in a significant reduction in the excretion of 3-methylthioparacetamol by urine, the glucuronide and sulphate.

  7. Therapeutic approaches targeting intestinal microflora in inflammatory bowel disease

    Institute of Scientific and Technical Information of China (English)

    Akira Andoh; Yoshihide Fujiyama

    2006-01-01

    Inflammatory bowel diseases, ulcerative colitis, and Crohn's disease, are chronic intestinal disorders of unknown etiology in which in genetically susceptible individuals, the mucosal immune system shows an aberrant response towards commensal bacteria.The gastrointestinal tract has developed ingenious mechanisms to coexist with its autologous microflora,but rapidly responds to invading pathogens and then returns to homeostasis with its commensal bacteria after the pathogenic infection is cleared. In case of disruption of this tightly-regulated homeostasis, chronic intestinal inflammation may be induced. Previous studies showed that some commensal bacteria are detrimental while others have either no influence or have a protective action. In addition, each host has a genetically determined response to detrimental and protective bacterial species. These suggest that therapeutic manipulation of imbalance of microflora can influence health and disease. This review focuses on new insights into the role of commensal bacteria in gut health and disease, and presents recent findings in innate and adaptive immune interactions. Therapeutic approaches to modulate balance of intestinal microflora and their potential mechanisms of action are also discussed.

  8. [Sensitivity to antibiotics monitoring of oral microflora in practically sound children and patients with chronic gastroduodenitis].

    Science.gov (United States)

    Gavrilova, O A; Davydov, B N; Chervinets, Iu V; Chervinets, V M

    2009-01-01

    Characteristic of oral microflora in practically sound children and patients with chronic gastroduodenitis was offered. Microflora singled out from ill persons differed by pathogenicity in previously opportunistic pathogenic forms. Monitoring of sensitivity and resistivity to antibiotics of the singled out microflora in practically sound children and ill with chronic gastroduodenitis was presented. When treating oral diseases one should take into account the high resistivity to antibiotics (especially to benzilpenicillin) of opportunistic pathogenic forms.

  9. Knockdown of midgut genes by dsRNA-transgenic plant-mediated RNA interference in the hemipteran insect Nilaparvata lugens.

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    Wenjun Zha

    Full Text Available BACKGROUND: RNA interference (RNAi is a powerful technique for functional genomics research in insects. Transgenic plants producing double-stranded RNA (dsRNA directed against insect genes have been reported for lepidopteran and coleopteran insects, showing potential for field-level control of insect pests, but this has not been reported for other insect orders. METHODOLOGY/PRINCIPAL FINDINGS: The Hemipteran insect brown planthopper (Nilaparvata lugens Stål is a typical phloem sap feeder specific to rice (Oryza sativa L.. To analyze the potential of exploiting RNAi-mediated effects in this insect, we identified genes (Nlsid-1 and Nlaub encoding proteins that might be involved in the RNAi pathway in N. lugens. Both genes are expressed ubiquitously in nymphs and adult insects. Three genes (the hexose transporter gene NlHT1, the carboxypeptidase gene Nlcar and the trypsin-like serine protease gene Nltry that are highly expressed in the N. lugens midgut were isolated and used to develop dsRNA constructs for transforming rice. RNA blot analysis showed that the dsRNAs were transcribed and some of them were processed to siRNAs in the transgenic lines. When nymphs were fed on rice plants expressing dsRNA, levels of transcripts of the targeted genes in the midgut were reduced; however, lethal phenotypic effects after dsRNA feeding were not observed. CONCLUSIONS: Our study shows that genes for the RNAi pathway (Nlsid-1 and Nlaub are present in N. lugens. When insects were fed on rice plant materials expressing dsRNAs, RNA interference was triggered and the target genes transcript levels were suppressed. The gene knockdown technique described here may prove to be a valuable tool for further investigations in N. lugens. The results demonstrate the potential of dsRNA-mediated RNAi for field-level control of planthoppers, but appropriate target genes must be selected when designing the dsRNA-transgenic plants.

  10. Investigation of the midgut structure and ultrastructure in Cimex lectularius and Cimex pipistrelli (Hemiptera: Cimicidae).

    Science.gov (United States)

    Rost-Roszkowska, M M; Vilimova, J; Włodarczyk, A; Sonakowska, L; Kamińska, K; Kaszuba, F; Marchewka, A; Sadílek, D

    2017-02-01

    Cimicidae are temporary ectoparasites, which means that they cannot obtain food continuously. Both Cimex species examined here, Cimex lectularius (Linnaeus 1758) and Cimex pipistrelli (Jenyns 1839), can feed on a non-natal host, C. lectularius from humans on bats, C. pipistrelli on humans, but never naturally. The midgut of C. lectularius and C. pipistrelli is composed of three distinct regions-the anterior midgut (AMG), which has a sack-like shape, the long tube-shaped middle midgut (MMG), and the posterior midgut (PMG). The different ultrastructures of the AMG, MMG, and PMG in both of the species examined suggest that these regions must fulfill different functions in the digestive system. Ultrastructural analysis showed that the AMG fulfills the role of storing food and synthesizing and secreting enzymes, while the MMG is the main organ for the synthesis of enzymes, secretion, and the storage of the reserve material. Additionally, both regions, the AMG and MMG, are involved in water absorption in the digestive system of both Cimex species. The PMG is the part of the midgut in which spherites accumulate. The results of our studies confirm the suggestion of former authors that the structure of the digestive tract of insects is not attributed solely to diet but to the basic adaptation of an ancestor.

  11. Effects of Periplocoside P from Periploca sepium on the Midgut Transmembrane Potential of Mythimna separata Larvae

    Science.gov (United States)

    Wang, YingYing; Qi, Zhijun; Qi, Meng; Hu, Zhaonong; Wu, Wenjun

    2016-01-01

    Periplocoside P (PSP) isolated from the root bark of Periploca sepium contains a pregnane glycoside skeleton and possesses high insecticidal properties. Preliminary studies indicated that PSP disrupts epithelial functions in the midgut of lepidopteran larvae. In the present study, we examined the effects of PSP on the apical and basolateral membrane voltages, Va and Vbl, respectively, of cells from (1) midguts isolated from the larvae of the oriental armyworm Mythimna separata that were in vitro incubated with toxins and (2) midguts isolated from M. separata larvae force-fed with PSP. We compared the effects of PSP with the effects of the Bacillus thuringiensis toxin Cry1Ab and inactive periplocoside E (PSE) on the midgut epithelial cells. The results showed that Va rapidly decreased in the presence of PSP in a time- and dose-dependent manner, similar to the effects of Cry1Ab. By contrast, PSE did not affect the Va and Vbl. Additionally, PSP did not influence the Vbl. Given these results, we speculate that PSP may modulate transport mechanisms at the apical membrane of the midgut epithelial cells by inhibiting the V-type H+ ATPase. PMID:27833169

  12. Histopathological Effects of the Protein Toxin from Xenorhabdus nematophila on the Midgut of Helicoverpa armigera

    Institute of Scientific and Technical Information of China (English)

    NANGONG Zi-yan; WANG Qin-ying; SONG Ping; YANG Jun; MAO Wen-jie

    2006-01-01

    Xenorhabdus nematophila HB310, which is highly virulent for many insects, is symbiotic with Steinernema carpocapsae HB310. Toxin Ⅱ was obtained using methods such as salting out and native-PAGE from the cells of X. Nematophila HB310. The histopathology of toxin Ⅱ on H. Armigera larvae was studied by dissecting an olefin slice of the midgut. The symptoms showed that the histopathology of the H. Armigera midgut was similar to that of other novel midgut-active toxins such as the δ-endotoxins from Bacillus thuringiensis, as well as Tca from Photorhabdus luminescens W14. The midgut tissues of H. Armigera fourth-instar larvae began to transform after the oral intake of the toxin Ⅱ over 6 h. First, the anterior region of the peritrophic membrane (PM) began to degrade followed by the elongation of the columnar cells.The epithelium decomposed gradually, and the midgut tissues were either loose or disordered. The PM disappeared after 12 h but reappeared after 72 h following transient or sublethal exposure to the toxin Ⅱ. Toxin Ⅱ also directly destroyed in vitro PMs of H. Armigera.

  13. Insect midgut carboxypeptidases with emphasis on S10 hemipteran and M14 lepidopteran carboxypeptidases.

    Science.gov (United States)

    Ferreira, C; Rebola, K G O; Cardoso, C; Bragatto, I; Ribeiro, A F; Terra, W R

    2015-04-01

    We compared the whole complement of midgut carboxypeptidases from 10 insects pertaining to five orders based on transcriptomes obtained by deep sequencing and biochemical data. Most of the carboxypeptidases were metallocarboxypeptidases from family M14, with carboxypeptidase A (CPA) predominating over carboxypeptidase B (CPB). They were found in all of the insects studied except for the hemipterans and a bruchid beetle. M14 carboxypeptidases were expressed only in the midgut of Spodoptera frugiperda (Lepidoptera). The most expressed CPA from this insect (SfCPA) was cloned, sequenced and expressed as a recombinant enzyme. This enzyme was used to generate antibodies used to demonstrate that SfCPA is secreted by an exocytic route. Serine carboxypeptidases from family S10 were found in all of the insects studied here. In S. frugiperda, they are expressed in all tissues besides the midgut, in accordance with their presumed lysosomal role. In the hemipteran Dysdercus peruvianus, S10 carboxypeptidases are expressed only in midgut, suggesting that they are digestive enzymes. This was confirmed by enzyme assays of midgut contents. Furthermore, the substrate specificity of D. peruvianus S10 carboxypeptidases are predicted to be one CPC (preferring hydrophobic residues) and one CPD (preferring basic residues), thus able to hydrolyse the peptides formed by their digestive cathepsin D and cathepsin L, respectively. The role of S10 carboxypeptidases in bruchid beetles are suggested to be the same as in hemipterans.

  14. Alterations in the Helicoverpa armigera midgut digestive physiology after ingestion of pigeon pea inducible leucine aminopeptidase.

    Directory of Open Access Journals (Sweden)

    Purushottam R Lomate

    Full Text Available Jasmonate inducible plant leucine aminopeptidase (LAP is proposed to serve as direct defense in the insect midgut. However, exact functions of inducible plant LAPs in the insect midgut remain to be estimated. In the present investigation, we report the direct defensive role of pigeon pea inducible LAP in the midgut of Helicoverpa armigera (Lepidoptera: Noctuidae and responses of midgut soluble aminopeptidases and serine proteinases upon LAP ingestion. Larval growth and survival was significantly reduced on the diets supplemented with pigeon pea LAP. Aminopeptidase activities in larvae remain unaltered in presence or absence of inducible LAP in the diet. On the contrary, serine proteinase activities were significantly decreased in the larvae reared on pigeon pea LAP containing diet as compared to larvae fed on diet without LAP. Our data suggest that pigeon pea inducible LAP is responsible for the degradation of midgut serine proteinases upon ingestion. Reduction in the aminopeptidase activity with LpNA in the H. armigera larvae was compensated with an induction of aminopeptidase activity with ApNA. Our findings could be helpful to further dissect the roles of plant inducible LAPs in the direct plant defense against herbivory.

  15. MicroRNA-regulation of Anopheles gambiae immunity to Plasmodium falciparum infection and midgut microbiota.

    Science.gov (United States)

    Dennison, Nathan J; BenMarzouk-Hidalgo, Omar J; Dimopoulos, George

    2015-03-01

    Invasion of the malaria vector Anopheles gambiae midgut by Plasmodium parasites triggers transcriptional changes of immune genes that mediate the antiparasitic defense. This response is largely regulated by the Toll and Immune deficiency (IMD) pathways. To determine whether A. gambiae microRNAs (miRNAs) are involved in regulating the anti-Plasmodium defense, we showed that suppression of miRNA biogenesis results in increased resistance to Plasmodium falciparum infection. In silico analysis of A. gambiae immune effector genes identified multiple transcripts with miRNA binding sites. A comparative miRNA microarray abundance analysis of P. falciparum infected and naïve mosquito midgut tissues showed elevated abundance of miRNAs aga-miR-989 and aga-miR-305 in infected midguts. Antagomir inhibition of aga-miR-305 increased resistance to P. falciparum infection and suppressed the midgut microbiota. Conversely, treatment of mosquitoes with an artificial aga-miR-305 mimic increased susceptibility to P. falciparum infection and resulted in expansion of midgut microbiota, suggesting that aga-miR-305 acts as a P. falciparum and gut microbiota agonist by negatively regulating the mosquito immune response. In silico prediction of aga-miR-305 target genes identified several anti-Plasmodium effectors. Our study shows that A. gambiae aga-miR-305 regulates the anti-Plasmodium response and midgut microbiota, likely through post-transcriptional modification of immune effector genes.

  16. Comparison of the faecal microflora of patients with ankylosing spondylitis and controls using molecular methods of analysis

    NARCIS (Netherlands)

    Stebbings, S; Munro, K; Simon, MA; Tannock, G; Highton, J; Harmsen, H; Welling, G; Seksik, P; Dore, J; Grame, G; Tilsala-Timisjarvi, A

    2002-01-01

    Objectives. To determine whether differences within the complex intestinal microflora can be demonstrated between patients with ankylosing spondylitis (AS) and healthy individuals. Methods. The composition of the faecal microflora of 15 ankylosing spondylitis patients and 15 matched controls was det

  17. Functional morphology of the midgut of a sandfly as compared to other hematophagous nematocera.

    Science.gov (United States)

    Rudin, W; Hecker, H

    1982-01-01

    The midgut epithelium of female Lutzomyia longipalpis was investigated by means of electron microscopic morphometry before and during blood digestion. Ultrastructure and cytological changes of the stomach cells upon blood feeding were generally similar to the ones described for Phlebotomus longipes (Gemetchu, 1974) and for mosquitoes (Hecker, 1977). In addition, the quantitative composition of the cells resembled the one of mosquitoes in many respects. Despite some morphological differences in the functional gut cytology, it can be admitted that, in general, digestive processes may run similarly in the midguts of sandflies and mosquitoes.

  18. Detection of heparin in the salivary gland and midgut of Aedes togoi.

    Science.gov (United States)

    Ha, Young-Ran; Oh, So-Ra; Seo, Eun-Seok; Kim, Bo-Heum; Lee, Dong-Kyu; Lee, Sang-Joon

    2014-04-01

    Mosquitoes secrete saliva that contains biological substances, including anticoagulants that counteract a host's hemostatic response and prevent blood clotting during blood feeding. This study aimed to detect heparin, an anticoagulant in Aedes togoi using an immunohistochemical detection method, in the salivary canal, salivary gland, and midgut of male and female mosquitoes. Comparisons showed that female mosquitoes contained higher concentrations of heparin than male mosquitoes. On average, the level of heparin was higher in blood-fed female mosquitoes than in non-blood-fed female mosquitoes. Heparin concentrations were higher in the midgut than in the salivary gland. This indicates presence of heparin in tissues of A. togoi.

  19. Effect of ceftobiprole on the normal human intestinal microflora.

    Science.gov (United States)

    Bäckström, Tobias; Panagiotidis, Georgios; Beck, Olof; Asker-Hagelberg, Charlotte; Rashid, Mamun-Ur; Weintraub, Andrej; Nord, Carl Erik

    2010-12-01

    Ceftobiprole is a new broad-spectrum pyrrolidinone cephem active against meticillin-resistant Staphylococcus aureus, vancomycin-resistant Enterococcus faecalis and Gram-negative bacteria such as Enterobacteriaceae and Pseudomonas spp. The purpose of the present study was to investigate the effect of administration of ceftobiprole on the normal intestinal microflora. Twelve healthy subjects (six males and six females) aged 20-31 years received ceftobiprole 500 mg by intravenous infusion every 8h for 7 days. Plasma samples were collected on Days -1, 1, 4, 7, 10, 14 and 21 for determination of drug concentration by biological and chemical methods. Faecal samples were collected on Days -1, 2, 4, 7, 10, 14 and 21. For analysis of the microflora, faecal specimens were cultured on non-selective and selective media. Different colony types were counted, isolated in pure culture and identified to genus level. All new colonising aerobic and anaerobic bacteria were tested for susceptibility to ceftobiprole. Plasma concentrations of ceftobiprole 10 min after completion of infusion were as follows: Day 1, 14.7-23.6 mg/L; Day 4, 15.9-24.5 mg/L; and Day 7, 15.9-23.9 mg/L. No ceftobiprole was detected in plasma on Days -1, 10, 14 and 21. No measurable concentrations of ceftobiprole were found in faeces on Days -1, 2, 4, 7, 10, 14 and 21. There were minor changes in the numbers of enteric bacteria, enterococci and Candida albicans and there were moderate changes in the numbers of bifidobacteria, lactobacilli, clostridia and Bacteroides spp. during the same period. No Clostridium difficile strains or toxins were found. No new colonising aerobic and anaerobic bacteria with ceftobiprole minimum inhibitory concentrations of ≥ 4 mg/L were found. Ceftobiprole had no significant ecological impact on the human intestinal microflora.

  20. The microflora of rainbow trout intestine : a comparison of traditional and molecular identification

    DEFF Research Database (Denmark)

    Spanggaard, Bettina; Huber, I.; Nielsen, J.

    2000-01-01

    The culturability of the intestinal microflora of 48 rainbow trout was detected by comparing direct microscopic counts (4',6-diamidino-2-phenylindole, DAPI) with plate counts (tryptone soya agar, TSA). In general, a high percentage (average 50%) of the microflora could be cultured. The counts...

  1. Differences in Peri-Implant Microflora Between Fully and Partially Edentulous Patients : A Systematic Review

    NARCIS (Netherlands)

    de Waal, Yvonne C.; Winkel, Edwin G.; Meijer, Henny J; Raghoebar, Gerry M.; van Winkelhoff, Arie J.

    2014-01-01

    Background: The current evidence suggests that the oral microflora differs between individuals who are fully edentulous (FES) and those who are partially edentulous (PES). It is unknown whether this leads to differences in peri-implant microflora when implants are installed. The aim of the study is

  2. Influence of hydrocolloidal silver nanoparticles on gastrointestinal microflora and morphology of enterocytes of quails

    DEFF Research Database (Denmark)

    Sawosz, Ewa; Binek, Marian; Grodzik, Marta

    2007-01-01

    killed and samples of duodenum and caeca microflora were collected. This initial investigation demonstrated that silver nanoparticles did not influence emphatically microflora of quail caecum; however, water containing 25 mg/kg of Ag-nano significantly increased the population of lactic acid bacteria...

  3. [Role of oral cavity anaerobic microflora in the pathogenesis of mandible traumatic osteomyelitis].

    Science.gov (United States)

    Ivaniushko, T P; Tumbinskaia, L V; Smirnov, A V; Balykin, R A

    2012-01-01

    The role of conditionally pathogenic microflora in the pathogenesis of lower jaw bone traumatic osteomyelites was investigated. The different groups of aerobic and anaerobic microorganisms were detected by real-time PCR. Role of conditionally pathogenic microflora in the pathogenesis of traumatic osteomielytis was shown.

  4. Impact of Deoxynivalenol on the Intestinal Microflora of Pigs

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    Philippe Fravalo

    2008-12-01

    Full Text Available Deoxynivalenol (DON, a mycotoxin produced by some Fusarium species, is a frequent contaminant of cereal. In the present study, 24 weanling piglets received either control feed or feed naturally contaminated with DON (2.8 mg/kg for four weeks. Consumption of contaminated feed significantly reduced the animal weight gain during the first week of the experiment, but had a moderate effect on cultivable bacteria in the pig intestine. By contrast, changes in the intestinal microflora were observed by Capillary Electrophoresis Single-Stranded Conformation Polymorphism (CE-SSCP in DON-exposed animals, suggesting an impact of this toxin on the dynamics of intestinal bacteria communities.

  5. Impact of salinomycin on the intestinal microflora of broiler chickens

    DEFF Research Database (Denmark)

    Johansen, Charlotte; Friis-Holm, Lotte Bjerrum; Pedersen, Karl

    2007-01-01

    Background: The ionophoric coccidiostat salinomycin is widely used in chicken feed. In the near future the use of ionophore coccidiostats may be banned as has been the case for other antimicrobial growth promoters. This study was conducted to examine the effect of salinomycin on Campylobacter....... Results: We found no effect of salinomycin on C. jejuni but salinomycin significantly affected the composition of the microflora. In addition, salinomycin significantly reduced the prevalence of Clostridium perfringens and we observed a significant increase (62%) in the mean body weight of salinomycin...

  6. Maternal vaginal microflora during pregnancy and the risk of asthma hospitalization and use of antiasthma medication in early childhood

    DEFF Research Database (Denmark)

    Benn, Christine Stabell; Thorsen, Poul; Jensen, Jørgen Skov;

    2002-01-01

    BACKGROUND: Infants with wheezing and allergic diseases have a microflora that differs from that of healthy infants. The fetus acquires microorganisms during birth when exposed to the maternal vaginal microflora. It is therefore conceivable that the maternal vaginal microflora might influence the...

  7. CHARACTERIZATION OF RABBITS MICROFLORA AFTER NATURAL PRODUCTS APPLICATION

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    Miroslava Kačániová

    2014-02-01

    Full Text Available The aim of this study was to compare the microbial species in caecum microflora of rabbit in control group against experimental group where epicatechin and patulin were applied. It is first study about caecum microflora study after epicatechin and patulin toxin application. In this study classical method was used for enumeration of bacteria . Enterococci were counted on Slanetz-Bartley agar (Biolife, Italy and incubated at 37 °C for 48-72 hours, lactobacilli were counted on MRS Lactobacillus agar (Biolife, Italy and incubated at 37 °C for 48-72 hours and coliforms bacteria were counted on MacConkey agar (Biolife, Italy and incubated at 37 °C for 24-48 hours. In our study the number of coliforms bacteria ranged between 3.78 to 4.45 log cfu.g-1, lactobacilli ranged between 3.85 to 5.00 log cfu.g-1 and enterococci ranged between 3.48 to 4.48 log cfu.g-1.

  8. Effect of special Hungarian probiotic kefir on faecal microflora

    Institute of Scientific and Technical Information of China (English)

    Mária Figler; Gyula Mózsik; Béla Schaffer; Beáta Gasztonyi; Pongrác (A)cs; Béla Szili; Regina Rab; Sándor Szakály

    2006-01-01

    AIM: To investigate the effect of a four-week consumption of a special Hungarian probiotic agent (Biofir(R)) on the faecal microflora in human healthy subjects.METHODS: The effect of Biofir(R) with 106/cm3 initial germs on the faecal microflora was studied in 120healthy volunteers (71 females, 49 males). The traditional Russian type kefir was used as control. The various germ groups and pH values were determined in wk 2, 4and 6.RESULTS: The number of all microbes increased during the 4-week probiotic treatment. The number of microbes increased 4.3-fold in the control group and 6.8-fold in Biofir-treated group. The probiotic kefir caused multiplication of the probiotic flora, meanwhile the undesired bacteria multiplied in the control group. No significant change of pH values of the faeces was found in both groups.CONCLUSION: The Hungarian probiotic kefir (Biofir(R)) is capable of promoting multiplication of probiotic bacterial flora in the large bowel.

  9. Intestinal microflora in rats with ischemia/reperfusion liver injury

    Institute of Scientific and Technical Information of China (English)

    XING Hui-chun; LI Lan-juan; XU Kai-jin; SHEN Tian; CHEN Yun-bo; SHENG Ji-fang; YU Yun-song; CHEN Ya-gang

    2005-01-01

    Objectives: To investigate the intestinal microflora status related to ischemia/reperfusion (I/R) liver injury and explore the possible mechanism. Methods: Specific pathogen free grade Sprague-Dawley rats were randomized into three groups: Control group (n=8), sham group (n=6) and I/R group (n=10). Rats in the control group did not receive any treatment, rats in the I/R group were subjected to 20 min of liver ischemia, and rats in the sham group were only subjected to sham operation. Twenty-two hours later, the rats were sacrificed and liver enzymes and malondialdehyde (MDA), superoxide dismutase (SOD), serum endotoxin,intestinal bacterial counts, intestinal mucosal histology, bacterial translocation to mesenteric lymph nodes, liver, spleen, and kidney were studied. Results: Ischemia/reperfusion increased liver enzymes, MDA, decreased SOD, and was associated with plasma endotoxin elevation in the I/R group campared to those in the sham group. Intestinal Bifidobacteria and Lactobacilli decreased and intestinal Enterobacterium and Enterococcus, bacterial translocation to kidney increased in the I/R group compared to the sham group. Intestinal microvilli were lost, disrupted and the interspace between cells became wider in the I/R group.Conclusion: I/R liver injury may lead to disturbance of intestinal microflora and impairment of intestinal mucosal barrier function,which contributes to endotoxemia and bacterial translocation to kidney.

  10. Transcriptome of the gypsy moth (Lymantria dispar) larval midgut in response to infection by Bacillus thuringiensis

    Science.gov (United States)

    Transcriptomic profiles of the lepidopteran insect pest Lymantria dispar (gypsy moth) were characterized in the larval midgut in response to infection by the biopesticide Bacillus thuringiensis kurstaki. RNA-Seq approaches were used to define a set of 49,613 assembled transcript sequences, of which...

  11. Characterization of Plasmodium developmental transcriptomes in Anopheles gambiae midgut reveals novel regulators of malaria transmission.

    Science.gov (United States)

    Akinosoglou, Karolina A; Bushell, Ellen S C; Ukegbu, Chiamaka Valerie; Schlegelmilch, Timm; Cho, Jee-Sun; Redmond, Seth; Sala, Katarzyna; Christophides, George K; Vlachou, Dina

    2015-02-01

    The passage through the mosquito is a major bottleneck for malaria parasite populations and a target of interventions aiming to block disease transmission. Here, we used DNA microarrays to profile the developmental transcriptomes of the rodent malaria parasite Plasmodium berghei in vivo, in the midgut of Anopheles gambiae mosquitoes, from parasite stages in the midgut blood bolus to sporulating oocysts on the basal gut wall. Data analysis identified several distinct transcriptional programmes encompassing genes putatively involved in developmental processes or in interactions with the mosquito. At least two of these programmes are associated with the ookinete development that is linked to mosquito midgut invasion and establishment of infection. Targeted disruption by homologous recombination of two of these genes resulted in mutant parasites exhibiting notable infection phenotypes. GAMER encodes a short polypeptide with granular localization in the gametocyte cytoplasm and shows a highly penetrant loss-of-function phenotype manifested as greatly reduced ookinete numbers, linked to impaired male gamete release. HADO encodes a putative magnesium phosphatase with distinctive cortical localization along the concave ookinete periphery. Disruption of HADO compromises ookinete development leading to significant reduction of oocyst numbers. Our data provide important insights into the molecular framework underpinning Plasmodium development in the mosquito and identifies two genes with important functions at initial stages of parasite development in the mosquito midgut.

  12. Kazal-type serine proteinase inhibitors in the midgut of Phlebotomus papatasi

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    Leah Theresa Sigle

    2013-09-01

    Full Text Available Sandflies (Diptera: Psychodidae are important disease vectors of parasites of the genus Leishmania, as well as bacteria and viruses. Following studies of the midgut transcriptome of Phlebotomus papatasi, the principal vector of Leishmania major, two non-classical Kazal-type serine proteinase inhibitors were identified (PpKzl1 and PpKzl2. Analyses of expression profiles indicated that PpKzl1 and PpKzl2 transcripts are both regulated by blood-feeding in the midgut of P. papatasi and are also expressed in males, larva and pupa. We expressed a recombinant PpKzl2 in a mammalian expression system (CHO-S free style cells that was applied to in vitro studies to assess serine proteinase inhibition. Recombinant PpKzl2 inhibited α-chymotrypsin to 9.4% residual activity and also inhibited α-thrombin and trypsin to 33.5% and 63.9% residual activity, suggesting that native PpKzl2 is an active serine proteinase inhibitor and likely involved in regulating digestive enzymes in the midgut. Early stages of Leishmania are susceptible to killing by digestive proteinases in the sandfly midgut. Thus, characterising serine proteinase inhibitors may provide new targets and strategies to prevent transmission of Leishmania.

  13. Two essential peritrophic matrix proteins mediate matrix barrier functions in the insect midgut.

    Science.gov (United States)

    Agrawal, Sinu; Kelkenberg, Marco; Begum, Khurshida; Steinfeld, Lea; Williams, Clay E; Kramer, Karl J; Beeman, Richard W; Park, Yoonseong; Muthukrishnan, Subbaratnam; Merzendorfer, Hans

    2014-06-01

    The peritrophic matrix (PM) in the midgut of insects consists primarily of chitin and proteins and is thought to support digestion and provide protection from abrasive food particles and enteric pathogens. We examined the physiological roles of 11 putative peritrophic matrix protein (PMP) genes of the red flour beetle, Tribolium castaneum (TcPMPs). TcPMP genes are differentially expressed along the length of the midgut epithelium of feeding larvae. RNAi of individual PMP genes revealed no abnormal developmental phenotypes for 9 of the 11 TcPMPs. However, RNAi for two PMP genes, TcPMP3 and TcPMP5-B, resulted in depletion of the fat body, growth arrest, molting defects and mortality. In situ permeability assays after oral administration of different-sized FITC-dextran beads demonstrated that the exclusion size of the larval peritrophic matrix (PM) decreases progressively from >2 MDa to RNAi for TcPMP3 and TcPMP5-B, these dextrans penetrated the epithelium of the median midgut, indicating loss of structural integrity and barrier function of the larval PM. In contrast, RNAi for TcPMP5-B, but not RNAi for TcPMP3, resulted in breakdown of impermeability to 4 and 40 kDa dextrans in the PM of the posterior midgut. These results suggest that specific PMPs are involved in the regulation of PM permeability, and that a gradient of barrier function is essential for survival and fat body maintenance.

  14. Alterations in the fat body and midgut of Culex quinquefasciatus larvae following exposure to different insecticides.

    Science.gov (United States)

    Alves, Stênio Nunes; Serrão, José Eduardo; Melo, Alan Lane

    2010-08-01

    This study describes morphological alterations in the fat body and midgut of Culex quinquefasciatus larvae following exposure to different insecticides. To this end, both third and fourth instars of C. quinquefasciatus larvae were exposed for 30 and 60 min to organophosphate (50 ppb), pyrethroids (20 and 30 ppb), and avermectin derivates (1.5 and 54 ppb). Following incubation, pH measurements of the larvae gut were recorded. The fat body and midgut were also analyzed by light and transmission electron microscopy. These studies demonstrate a decrease in the pH of the larvae anterior midgut following exposure to all of the tested insecticides. Histochemical tests revealed a strong reaction for neutral lipids in the control group and a marked decrease in the group exposed to cypermethrin. Furthermore, a weak reaction with acidic lipids in larvae exposed to deltamethrin, temephos, ivermectin and abamectin was also observed. Insecticide-exposed larvae also exhibited cytoplasm granule differences, relative to control larvae. Finally, we noted a small reduction in microvilli size in the apex of digestive cells, although vesicles were found to be present. The destructive changes in the larvae were very similar regardless of the type of insecticide analyzed. These data suggest that alterations in the fat body and midgut are a common response to cellular intoxication.

  15. Apoptosis and necrosis during the circadian cycle in the centipede midgut.

    Science.gov (United States)

    Rost-Roszkowska, M M; Chajec, Ł; Vilimova, J; Tajovský, K

    2016-07-01

    Three types of cells have been distinguished in the midgut epithelium of two centipedes, Lithobius forficatus and Scolopendra cingulata: digestive, secretory, and regenerative cells. According to the results of our previous studies, we decided to analyze the relationship between apoptosis and necrosis in their midgut epithelium and circadian rhythms. Ultrastructural analysis showed that these processes proceed in a continuous manner that is independent of the circadian rhythm in L. forficatus, while in S. cingulata necrosis is activated at midnight. Additionally, the description of apoptosis and necrosis showed no differences between males and females of both species analyzed. At the beginning of apoptosis, the cell cytoplasm becomes electron-dense, apparently in response to shrinkage of the cell. Organelles such as the mitochondria, cisterns of endoplasmic reticulum transform and degenerate. Nuclei gradually assume lobular shapes before the apoptotic cell is discharged into the midgut lumen. During necrosis, however, the cytoplasm of the cell becomes electron-lucent, and the number of organelles decreases. While the digestive cells of about 10 % of L. forficatus contain rickettsia-like pathogens, the corresponding cells in S. cingulata are free of rickettsia. As a result, we can state that apoptosis in L. forficatus is presumably responsible for protecting the organism against infections, while in S. cingulata apoptosis is not associated with the elimination of pathogens. Necrosis is attributed to mechanical damage, and the activation of this process coincides with proliferation of the midgut regenerative cells at midnight in S. cingulata.

  16. Perimicrovillar membrane assembly: the fate of phospholipids synthesised by the midgut of Rhodnius prolixus

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    Paula Rego Bittencourt-Cunha

    2013-06-01

    Full Text Available In this study, we describe the fate of fatty acids that are incorporated from the lumen by the posterior midgut epithelium of Rhodnius prolixus and the biosynthesis of lipids. We also demonstrate that neutral lipids (NL are transferred to the haemolymphatic lipophorin (Lp and that phospholipids remain in the tissue in which they are organised into perimicrovillar membranes (PMMs. 3H-palmitic acid added at the luminal side of isolated midguts of R. prolixus females was readily absorbed and was used to synthesise phospholipids (80% and NL (20%. The highest incorporation of 3H-palmitic acid was on the first day after a blood meal. The amounts of diacylglycerol (DG and triacylglycerol synthesised by the tissue decreased in the presence of Lp in the incubation medium. The metabolic fates of 3H-lipids synthesised by the posterior midgut were followed and it was observed that DG was the major lipid released to Lp particles. However, the majority of phospholipids were not transferred to Lp, but remained in the tissue. The phospholipids that were synthesised and accumulated in the posterior midgut were found to be associated with Rhodnius luminal contents as structural components of PMMs.

  17. Transcriptional characteristics of gene expression in the midgut of domestic silkworms (Bombyx mori) exposed to phoxim.

    Science.gov (United States)

    Gu, Z Y; Sun, S S; Wang, Y H; Wang, B B; Xie, Y; Ma, L; Wang, J M; Shen, W D; Li, B

    2013-01-01

    Silkworm (Bombyx mori) is not only an economically important insect but also a model system for lepidoptera. As a vital organ of digestion and nutrient absorption, the midgut of insects also serves as the first physiological barrier to chemical pesticides. In this study, microarray was performed to profile the gene expression changes in the midgut of silkworms exposed to phoxim. After 24h of phoxim exposure (4.0μg/mL), 266 genes displayed at least 2.0-fold changes in expression levels. Among them, 192 genes were up-regulated, and 74 genes were down-regulated. The most significant changes were 14.88-fold up-regulation and 23.36-fold down-regulation. According to gene ontology annotation and pathway analysis, differentially expressed genes were mainly classified into different groups based on their potential involvements in detoxification, immunne response, stress response, energy metabolism and transport. Particularly, the transcription levels of detoxification-related genes were up-regulated, such as cytochrome P450s, esterases and glutathione-S-transferase (GST), indicating increased detoxification activity in the midgut. Our study provides new insights into the molecular mechanism of pesticide metabolism in the midgut of insects, which may promote the development of highly efficient insecticides.

  18. Effects of heavy metal accumulation on the midgut gland in a terrestrial isopod, Porcellio scaber

    Energy Technology Data Exchange (ETDEWEB)

    Szlavecz, K.; Komueves, L.G.; Gueth, S.

    1986-01-01

    The effects of lead and cadmium on the lysosomal enzyme activity, the protein content, and the ultrastructure of the midgut gland, which plays a central role in the metabolism of isopods, were studied. Hornbeam litter was sprayed with Pb acetate or Cd chloride solutions of different concentrations. The animals were fed with the contaminated leaf litter for one or two months. Leaf litter sprayed with distilled water served as control. X-ray fluorescence analysis showed considerable heavy metal accumulation in the whole body of the experimental animals. Total protein content of the midgut gland of the treated isopods was approximately 60% less than that of the controls. However, the total activities of the lysosomal enzymes (acid phosphatase, acid ..beta..-galactosidase, acid glucosidase) were not affected by the treatments, meaning, that specific activities of these enzymes increased. Electron microscopic investigation of the midgut gland revealed characteristic ultrastructural changes in the different cell types. The amount of glycogen and the number of lipid droplets and secretory granules decreased, whereas an increase in the number of autophagic vacuoles and large secondary lysosomes was observed. The study indicates, that heavy metals ingested with food are not only stored in the midgut gland, but they influence the ultrastructure and functions of its cells, as well.

  19. Midgut epithelium in molting silkworm: A fine balance among cell growth, differentiation, and survival.

    Science.gov (United States)

    Franzetti, Eleonora; Casartelli, Morena; D'Antona, Paola; Montali, Aurora; Romanelli, Davide; Cappellozza, Silvia; Caccia, Silvia; Grimaldi, Annalisa; de Eguileor, Magda; Tettamanti, Gianluca

    2016-07-01

    The midgut of insects has attracted great attention as a system for studying intestinal stem cells (ISCs) as well as cell death-related processes, such as apoptosis and autophagy. Among insects, Lepidoptera represent a good model to analyze these cells and processes. In particular, larva-larva molting is an interesting developmental phase since the larva must deal with nutrient starvation and its organs are subjected to rearrangements due to proliferation and differentiation events. Several studies have analyzed ISCs in vitro and characterized key factors involved in their division and differentiation during molt. However, in vivo studies performed during larva-larva transition on these cells, and on the whole midgut epithelium, are fragmentary. In the present study, we analyzed the larval midgut epithelium of the silkworm, Bombyx mori, during larva-larva molting, focusing our attention on ISCs. Moreover, we investigated the metabolic changes that occur in the epithelium and evaluated the intervention of autophagy. Our data on ISCs proliferation and differentiation, autophagy activation, and metabolic and functional activities of the midgut cells shed light on the complexity of this organ during the molting phase.

  20. Midgut transcriptome profiling of Anoplophora glabripennis, a lignocellulose degrading Cerambycid beetle

    Science.gov (United States)

    Background: Wood-feeding insects often work in collaboration with microbial symbionts to degrade lignin biopolymers and release glucose and other fermentable sugars from recalcitrant plant cell wall carbohydrates, including cellulose and hemicellulose. Here, we present the midgut transcriptome of la...

  1. Proteolytic activity in the midgut of Ectomyelois ceratoniae Zeller (Lepidoptera: Pyralidae, Pomegranate carob moth

    Directory of Open Access Journals (Sweden)

    M Ranjbar

    2011-07-01

    Full Text Available In this study, the proteolytic activity in the midgut of Ectomyelois ceratoniae as the major pest of pomegranate was investigated to find nature of specific proteases and their properties for adopting possible pest management procedure. It was found that fourth and fifth instar larvae had the highest proteolytic activity as well as specific proteinases including, elastase, trypsin-like, chymotrypsin-like and two exopeptidases. The optimal pH of general protease was 10 and 9 for azocasein, casein and hemoglobin as substrate. The optimal temperature of the total proteolytic activity in the midgut of E. ceratoniae was found 30 and 35 °C by using azocasein and casein as substrates, respectively. In case of hemoglobin, the enzyme showed the highest enzymatic activity at temperatures from 15 to 35 °C. There was no enhancement in the proteolytic activity by using different cations but SDS, citric acid and mercaptoetahnol significantly decreased the proteolytic activity in the midgut of E. ceratoniae. Using specific proteolytic inhibitors including PMSF, TLCK, TPCK, E-64, DTT and phenanthroline revealed presence of serine proteases as the major proteases in the midgut of E. ceratoniae.

  2. Localization of two post-proline cleaving peptidases in the midgut of Tenebrio molitor larvae

    Science.gov (United States)

    Two soluble post-proline cleaving peptidase activities, PPCP1 and PPCP2, were demonstrated in the midgut of Tenebrio molitor larvae with the substrate benzyloxycarbonyl-L-alanyl-L-proline p-nitroanilide. Both activities were serine peptidases. PPCP1 was active in acidic buffers, with maximum activit...

  3. Characterization of proteinases from the midgut of Rhipicephalus (Boophilus microplus involved in the generation of antimicrobial peptides

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    Craik Charles S

    2010-07-01

    Full Text Available Abstract Background Hemoglobin is a rich source of biologically active peptides, some of which are potent antimicrobials (hemocidins. A few hemocidins have been purified from the midgut contents of ticks. Nonetheless, how antimicrobials are generated in the tick midgut and their role in immunity is still poorly understood. Here we report, for the first time, the contribution of two midgut proteinases to the generation of hemocidins. Results An aspartic proteinase, designated BmAP, was isolated from the midgut of Rhipicephalus (Boophilus microplus using three chromatographic steps. Reverse transcription-quantitative polymerase chain reaction revealed that BmAP is restricted to the midgut. The other enzyme is a previously characterized midgut cathepsin L-like cysteine proteinase designated BmCL1. Substrate specificities of native BmAP and recombinant BmCL1 were mapped using a synthetic combinatorial peptide library and bovine hemoglobin. BmCL1 preferred substrates containing non-polar residues at P2 subsite and polar residues at P1, whereas BmAP hydrolysed substrates containing non-polar amino acids at P1 and P1'. Conclusions BmAP and BmCL1 generate hemocidins from hemoglobin alpha and beta chains in vitro. We postulate that hemocidins may be important for the control of tick pathogens and midgut flora.

  4. Action of Douglas Fir Tussock Moth Larvae and Their Microflora on Dietary Terpenes

    OpenAIRE

    Andrews, R E; Spence, K. D.

    1980-01-01

    A single type of bacterium, tentatively identified as a member of the genus Bacillus, was isolated from 2 of 20 midguts of Douglas fir tussock moth larvae being fed a diet of fir needles. No bacteria could be isolated from most midguts. Although spherically shaped bodies were present in the food bolus, these bodies, if microorganisms, could not be distinguished from spherical bodies associated with the plant tissue. The Douglas fir tussock moth dietary terpenes were altered during their passa...

  5. Midgut Transcriptome of the Cockroach Periplaneta americana and Its Microbiota: Digestion, Detoxification and Oxidative Stress Response.

    Directory of Open Access Journals (Sweden)

    Jianhua Zhang

    Full Text Available The cockroach, Periplaneta americana, is an obnoxious and notorious pest of the world, with a strong ability to adapt to a variety of complex environments. However, the molecular mechanism of this adaptability is mostly unknown. In this study, the genes and microbiota composition associated with the adaptation mechanism were studied by analyzing the transcriptome and 16S rDNA pyrosequencing of the P. americana midgut, respectively. Midgut transcriptome analysis identified 82,905 unigenes, among which 64 genes putatively involved in digestion (11 genes, detoxification (37 genes and oxidative stress response (16 genes were found. Evaluation of gene expression following treatment with cycloxaprid further revealed that the selected genes (CYP6J1, CYP4C1, CYP6K1, Delta GST, alpha-amylase, beta-glucosidase and aminopeptidase were upregulated at least 2.0-fold at the transcriptional level, and four genes were upregulated more than 10.0-fold. An interesting finding was that three digestive enzymes positively responded to cycloxaprid application. Tissue expression profiles further showed that most of the selected genes were midgut-biased, with the exception of CYP6K1. The midgut microbiota composition was obtained via 16S rDNA pyrosequencing and was found to be mainly dominated by organisms from the Firmicutes phylum, among which Clostridiales, Lactobacillales and Burkholderiales were the main orders which might assist the host in the food digestion or detoxification of noxious compounds. The preponderant species, Clostridium cellulovorans, was previously reported to degrade lignocellulose efficiently in insects. The abundance of genes involved in digestion, detoxification and response to oxidative stress, and the diversity of microbiota in the midgut might provide P. americana high capacity to adapt to complex environments.

  6. NASAL MUCOUS MEMBRANE MICROFLORA IN PATIENTS WITH POLYPOUS RHINOSINUSITIS

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    O. A. Kolenchukova

    2016-01-01

    Full Text Available Nasal mucous membrane microbiocenosis Research amongst patients with polypous rhinosinusitis is conducted. Patients with polypous rhinosinusitis (PRS, n = 58 aged from 18 till 64 years and group of control (n = 156. For an microflora assessment of nasal mucous membrane during an exacerbation of a disease carried out crops of microorga nisms on nutrient differential and diagnostic agars. When studying the microflora received from nasal mucous membrane 407 cultures of microorganisms at PRS were revealed. In control group of 174 microorganisms cultures are revealed. Among isolates were established 6 genera of 9 species of bacteria at PRS against 6 genera and 8 species in group of control. Microflora quantitative structure research of nasal mucous membrane at PRS of rather control group considerable prevalence of the microorganisms belonging to the sorts Staphylococcus and Streptococcus, and also to Enterobacteriaceae was revealed. It is also revealed increase S. pneumoniae. When determining specific accessory of the microorganisms relating to the Staphylococcus genera in PRS group concerning control the increase in total number of the strains of S. aureus relating to coagulase-positive and S. haemolyticus, S. epidermidis relating to coagulase-negative staphylococcus was established. A big variety the coagulase-negative of Staphylococcus is also revealed: S. epidermidis, S. haemolyticus, S. hominis, S. cohnii, S. capitis, S. hyicius and S. xylosus. In control group of such types as S. capitis and S. hyicius it isn’t revealed. Thus at a polypous rhinosinusitis nasal mucous membrane the expressed dysbacteriosis takes place. The analysis of frequency of occurrence of the microorganisms belonging to different childbirth showed that in PRS group bacteria of the sort Streptococcus and this Nesseria, and also S. pneumoniae, S. pyogenes were most often allocated concerning control. The frequency prevalence analysis of the gene ra Staphylococcus related

  7. Analysis of changes in intestinal microflora of irradiated mice. [Gamma radiation

    Energy Technology Data Exchange (ETDEWEB)

    Mal' tsev, V.N.; Pinegin, B.V.; Korshunov, V.M.

    1977-01-01

    In experiments on 3 groups of CBA mice exposed to doses of 900, 600 and 300 R ..gamma..-rays, it was demonstrated that the integral severity of post-radiation microflora in the intestine can be determined by means of information index h, which takes into consideration all changes occurring in different representatives of the intestinal microflora. Differential analysis of the mechanisms of radioinduced changes in microflora indicates that it is based on a decrease in lactobacilli and increase in enterococcus, proteus, colibacillus and yeast in the small intestine, with increase in colibacillus, clostridia, proteus and enterococcus in the large intestine.

  8. [THE EVALUATION OF MICROBIOLOGICAL DISORDERS OF MICROFLORA OF OROPHARYNX AND INTESTINE USING MATHEMATICAL MODELING TECHNIQUE].

    Science.gov (United States)

    Zatevalov, A V; Elkova, E P; Afanasiev, S S; Aleshkin, A V; Mironov, A Yu; Gusarova, M P; Gudova, N V

    2016-02-01

    The analysis was applied to microflora of feces and oropharinx and concentration of volatile fatty acids in saliva from patients of consultative diagnostic center of G.N. Gabrichevskii Moscow research institute of epidemiology and microbiology. The computer classification program is developed on the basis of determining degree of microbiological disorders on the basis of received data and using artificial neural networks and discriminant analysis. The analysis established decreasing of probability of false classification in case of increasing of degree of microbiological disorders of microflora of intestine and absence of such a correlation for microbiological and metabolic disorders of microflora of intestine.

  9. Antimicrobial activity of herbs against Yersinia enterocolitica and mixed microflora

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    Shilpa SHARMA

    2016-12-01

    Full Text Available The present study aimed at developing herbal medicine against food borne pathogens, therefore the antimicrobial activity of four herbs viz. Arjuna (bark, Ashwagandha (roots, Puthkanda (leaves and Shalampanja (roots was checked. Aqueous, ethanolic and petroleum ether extracts of each herb were extracted and their antimicrobial activity against mixed microflora and against Yersinia enterocolitica was determined. Tetracycline and gentamicin were used as reference antibiotics. Arjuna extracts showed the highest antimicrobial potential against mixed population and Yersinia enterocolitica in comparison to Ashwagandha, Puthkanda and Shalampanja extracts. The antimicrobial activity of Arjuna aqueous extract was lower compared to gentamicin, but comparable to tetracycline. The minimum inhibitory concentration and minimum bactericidal concentration of aqueous extract of Arjuna showed the lowest values indicating that it is more effective in lower concentration of use. The antimicrobial activity of herbs showed the following trend Arjuna > Puthkanda > Shalampanja > Ashwagandha.

  10. [Characterization of the microflora of the small intestine (author's transl)].

    Science.gov (United States)

    Bernhardt, H; Knoke, M

    1980-03-01

    Normal and abnormal microflora of the upper small intestine was studied in 356 patients. Low counts are characteristic of normal microbial colonization (eubiosis), changes in quality and/or quantity are pathological (dysbiosis). The latter status is described as overgrowth syndrome. We found some types of dysbiosis. Prevalent was type Dys1 with the highest counts and the greatest variety of bacteria and yeasts. In contrast to this, type Dys2 showed higher germ counts of only one genus like coliforms (Dys2 Coli), streptococci (Dys2 Str.), lactobacilli (Dys2 L.) or yeasts (Dys2 Y.). In dysbiosis, we frequently saw bifidobacterium and bacteroides. Simultaneous sampling from stomach, duodenum, and jejunum indicated different modes of colonization of these parts (oral or fecal type).

  11. Oral microflora in preschool children attending a fluoride varnish program

    DEFF Research Database (Denmark)

    Anderson, Maria; Grindefjord, Margaret; Dahllöf, Göran

    2016-01-01

    hundred seven 3-year-old children were enrolled from a cohort of 3403 preschool children taking part in a community based oral health project. Two hundred sixty-three of them had attended caries-preventive program with semi-annual applications of a fluoride varnish since the age of 1 year (test group......BACKGROUND: To compare the oral microflora in preschool children attending a fluoride varnish program with a reference group receiving a standard oral health program without fluoride varnish applications. A second aim was to relate the microbial composition to the caries prevalence. METHODS: Five......) while 237 had received standard preventive care (reference group). Oral samples were collected with a sterile swab and analysed with checkerboard DNA-DNA hybridization using 12 pre-determined bacterial probes. Caries and background data were collected from clinical examinations and questionnaires...

  12. Interaction of gut microflora with tannins in feeds

    Science.gov (United States)

    Goel, Gunjan; Puniya, A. K.; Aguilar, C. N.; Singh, Kishan

    2005-11-01

    Tannins (hydrolyzable and condensed) are water-soluble polyphenolic compounds that exert antinutritional effects on ruminants by forming complexes with dietary proteins. They limit nitrogen supply to animals, besides inhibiting the growth and activity of ruminal microflora. However, some gastrointestinal microbes are able to break tannin-protein complexes while preferentially degrading hydrolyzable tannins (HTs). Streptococcus gallolyticus, Lonepinella koalarum and Selenomonas ruminantium are the dominant bacterial species that have the ability to degrade HTs. These tanninolytic microorganisms possess tannin-degrading ability and have developed certain mechanisms to tolerate tannins in feeds. Hence, selection of efficient tanninolytic microbes and transinoculation among animals for long-term benefits become areas of intensive interest. Here, we review the effects of tannins on ruminants, the existence and significance of tannin-degrading microorganisms in diverse groups of animals and the mechanisms that tannin-degrading microorganisms have developed to counter the toxic effects of tannin.

  13. Effects of intensity-modulated radiotherapy on human oral microflora.

    Science.gov (United States)

    Shao, Zi-Yang; Tang, Zi-Sheng; Yan, Chao; Jiang, Yun-Tao; Ma, Rui; Liu, Zheng; Huang, Zheng-Wei

    2011-01-01

    This study aimed to evaluate changes in the biodiversity of the oral microflora of patients with head and neck cancer treated with postoperative intensity-modulated radiotherapy (IMRT) or conventional radiotherapy (CRT). Pooled dental plaque samples were collected during the radiation treatment from patients receiving IMRT (n = 13) and CRT (n = 12). Denaturing gradient gel electrophoresis (DGGE) was used to analyze the temporal variation of these plaque samples. The stimulated and unstimulated salivary flow rates were also compared between IMRT and CRT patients. Reductions in the severity of hyposalivation were observed in IMRT patients compared with CRT patients. We also observed that the temporal stability of the oral ecosystem was significantly higher in the IMRT group (69.96 ± 7.82%) than in the CRT group (51.98 ± 10.45%) (P oral ecosystem than CRT.

  14. Effects of a skin neuropeptide (substance p on cutaneous microflora.

    Directory of Open Access Journals (Sweden)

    Lily Mijouin

    Full Text Available BACKGROUND: Skin is the largest human neuroendocrine organ and hosts the second most numerous microbial population but the interaction of skin neuropeptides with the microflora has never been investigated. We studied the effect of Substance P (SP, a peptide released by nerve endings in the skin on bacterial virulence. METHODOLOGY/PRINCIPAL FINDINGS: Bacillus cereus, a member of the skin transient microflora, was used as a model. Exposure to SP strongly stimulated the cytotoxicity of B. cereus (+553±3% with SP 10(-6 M and this effect was rapid (<5 min. Infection of keratinocytes with SP treated B. cereus led to a rise in caspase1 and morphological alterations of the actin cytoskeleton. Secretome analysis revealed that SP stimulated the release of collagenase and superoxide dismutase. Moreover, we also noted a shift in the surface polarity of the bacteria linked to a peel-off of the S-layer and the release of S-layer proteins. Meanwhile, the biofilm formation activity of B. cereus was increased. The Thermo unstable ribosomal Elongation factor (Ef-Tu was identified as the SP binding site in B. cereus. Other Gram positive skin bacteria, namely Staphylococcus aureus and Staphylococcus epidermidis also reacted to SP by an increase of virulence. Thermal water from Uriage-les-Bains and an artificial polysaccharide (Teflose® were capable to antagonize the effect of SP on bacterial virulence. CONCLUSIONS/SIGNIFICANCE: SP is released in sweat during stress and is known to be involved in the pathogenesis of numerous skin diseases through neurogenic inflammation. Our study suggests that a direct effect of SP on the skin microbiote should be another mechanism.

  15. MICROFLORA OF BEET SUGAR PRODUCTION: PROBLEMS AND SOLUTIONS

    Directory of Open Access Journals (Sweden)

    N. G. Kulneva

    2014-01-01

    Full Text Available Summary. Sugar beet is one of the strategic crops for food safety of Russia. The lack of specialized warehouse for harvest does not provide storage of roots for a long time. In the case of a thaw roots that have been defrosted unsuitable for processing. Beet and products of its processing is a good object for the development of microorganisms. Permanent microflora of sugar production are: Bacillus subtilis, Clostridium perfringes, Leuconostoc dextranicum, Torula alba, Pseudomonas fluorescens, Sarcina lutea and other kinds of microorganisms, leading to a problem processing of beet root and reduced quality of sugar. The most dangerous is the slimy bacteriosis is a bacterial disease beet caused by heterofermentative cocci of Leuconostoc (Leuconostoc mesenteroides, L. dextranicum. Product of the vital activity of microorganisms is dextran, which is synthesized from sucrose as a result of dextrany or mucous fermentation and leads to significant technological problems in processing of infected beet. Improving the efficiency of sugar production is connected with decrease in loss of quality of raw material preparation, storing and processing of sugar beet. At sugar plants use a variety of drugs that suppress the growth of pathogenic microflora, but there comes a rapid adaptation of microorganisms, therefore there is a need to implement new products to prevent damage to roots and improve the quality of produced sugar. To resolve this problem experimentally selected bactericidal drug, defined its rational concentration and conditions for the use in sugar beet production. The use of antibacterial drug in the process of extraction allows to increase the purity of diffusion juice 1.3 %, reduce the protein content in it (12.5 %; with the purity of the pure juice increases by 1.1 %, its color index is reduced by 44.7 %.

  16. The oral microflora in obesity and type-2 diabetes

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    Edward Shillitoe

    2012-10-01

    Full Text Available Background: Type 2 diabetes mellitus (T2DM is prevalent in people with obesity. It has been proposed that these conditions are related to specific features of the microflora of the mouth and lower gastrointestinal (GI tract. Hyperglycemia often resolves quickly after Roux-en-Y gastric bypass (RYGB but the role of the GI microflora cannot be examined easily because of reduced intestinal mobility. We propose that the study of microorganisms present in the mouth of patients undergoing RYGB will contribute to our understanding of the role of bacteria in the pathogenesis of T2DM. Objective: To conduct a feasibility study to examine differences in oral microbes in obese patients with and without T2DM and to determine whether it is feasible to measure changes after gastric bypass surgery. Methods: Individuals with morbid obesity (n=29, of whom 13 had T2DM, were studied. Oral rinses, stool samples, and blood samples were obtained before RYGB, and oral rinses and blood samples were obtained at 2 and 12 weeks postsurgery. Results: Prior to surgery, participants with T2DM had slightly higher total levels of oral bacteria than those without diabetes. Those with HbA1c > 6.5% had rather lower levels of Bifidobacteria in the mouth and stool. At 2 weeks post-RYGB, patients with T2DM were able to reduce or discontinue their hypoglycemic medications. Stool samples could not be obtained but oral rinses were readily available. The levels of oral Bifidobacteria had increased tenfold and levels of circulating endotoxin and tumor necrosis factor-alpha had decreased. Conclusions: The study of oral bacteria before and after RYGB is feasible and should be tested in larger patient populations to increase our understanding of the role of microorganisms in the pathogenesis of obesity and T2DM.

  17. Effects of Anti-Mosquito Salivary Glands and Deglycosylated Midgut Antibodies of Anopheles stephensi on Fecundity and Longevity

    Directory of Open Access Journals (Sweden)

    H Mohammadzadeh Hajipirloo

    2005-09-01

    Full Text Available With the aim of controlling malaria by reducing vector population, the effects of antibodies produced against salivary glands and deglycosylated midgut antigens of Anopheles stephensi mosquitoes on fecundity and longevity of the same species were tested. Three deglycosylated preparations of midgut and two preparations of salivary glands were produced, conjugated with aluminum hydroxide gel, and subcutaneously injected to shoulders of TO (Turner Out-bred mice. After 4 immunizations and assurance of enough antibody production against utilized antigenic suspensions, effects of blood feeding on immunized and control mice were assayed. Insoluble preparation of midgut showed the strongest effect with 23.5% reduction in egg laying, and increasing death rate of vectors in third day after feeding. No significant reduction in fecundity or survivorship was seen with other preparations. Anopheles midgut insoluble antigens are potential candidates for designing vaccines against malaria vectors and further investigations need to be done to find effective antigens and the best way of their use.

  18. Do malaria ookinete surface proteins P25 and P28 mediate parasite entry into mosquito midgut epithelial cells?

    Directory of Open Access Journals (Sweden)

    Ranford-Cartwright Lisa C

    2005-02-01

    Full Text Available Abstract Background P25 and P28 are related ookinete surface proteins highly conserved throughout the Plasmodium genus that are under consideration as candidates for inclusion in transmission-blocking vaccines. Previous research using transgenic rodent malaria parasites lacking P25 and P28 has demonstrated that these proteins have multiple partially redundant functions during parasite infection of the mosquito vector, including an undefined role in ookinete traversal of the mosquito midgut epithelium, and it has been suggested that, unlike wild-type parasites, Dko P25/P28 parasites migrate across the midgut epithelium via an intercellular, rather than intracellular, route. Presentation of the hypothesis This paper presents an alternative interpretation for the previous observations of Dko P25/P28 parasites, based upon a recently published model of the route of ookinete invasion across the midgut epithelium. This model claims ookinete invasion is intracellular, with entry occurring through the lateral apical plasma membrane of midgut epithelial cells, and is associated with significant invagination of the midgut epithelium localised at the site of parasite penetration. Following this model, it is hypothesized that: (1 a sub-population of Dko P25/P28 ookinetes invaginate, but do not penetrate, the apical surface of the midgut epithelium and thus remain within the midgut lumen; and (2 another sub-population of Dko P25/P28 parasites successfully enters and migrates across the midgut epithelium via an intracellular route similar to wild-type parasites and subsequently develops into oocysts. Testing the hypothesis These hypotheses are tested by showing how they can account for previously published observations and incorporate them into a coherent and consistent explanatory framework. Based upon these hypotheses, several quantitative predictions are made, which can be experimentally tested, about the relationship between the densities of invading Dko P

  19. Larval midgut modifications associated with Bti resistance in the yellow fever mosquito using proteomic and transcriptomic approaches

    OpenAIRE

    Tetreau Guillaume; Bayyareddy Krishnareddy; Jones Christopher M; Stalinski Renaud; Riaz Muhammad A; Paris Margot; David Jean-Philippe; Adang Michael J; Després Laurence

    2012-01-01

    Abstract Background Bacillus thuringiensis var. israelensis (Bti) is a natural larval mosquito pathogen producing pore-forming toxins targeting the midgut of Diptera larvae. It is used worldwide for mosquito control. Resistance mechanisms of an Aedes aegypti laboratory strain selected for 30 generations with field-collected leaf litter containing Bti toxins were investigated in larval midguts at two levels: 1. gene transcription using DNA microarray and RT-qPCR and 2. differential expression ...

  20. Biotransformation of aflatoxin B1 and its conjugated metabolites by rat gastrointestinal microfloras.

    OpenAIRE

    1981-01-01

    Rat cecal microflora from high- and low-fiber-fed animals hydrolyzed aflatoxin conjugates to metabolites indistinguishable from aflatoxin B1 and aflatoxin P1, but aflatoxicol was not a transformation product.

  1. Gastric acid reduction leads to an alteration in lower intestinal microflora

    Energy Technology Data Exchange (ETDEWEB)

    Kanno, Takayuki [Department of Gastroenterology, Wakayama Medical University, 811-1 Kimiidera, Wakayama-city, Wakayama 641-0012 (Japan); Matsuki, Takahiro [Yakult Central Institute for Microbiological Research, Tokyo (Japan); Oka, Masashi; Utsunomiya, Hirotoshi [Department of Gastroenterology, Wakayama Medical University, 811-1 Kimiidera, Wakayama-city, Wakayama 641-0012 (Japan); Inada, Kenichi [First Department of Pathology, Fujita Health University School of Medicine, Aichi (Japan); Magari, Hirohito; Inoue, Izumi; Maekita, Takao; Ueda, Kazuki; Enomoto, Shotaro; Iguchi, Mikitaka; Yanaoka, Kimihiko; Tamai, Hideyuki [Department of Gastroenterology, Wakayama Medical University, 811-1 Kimiidera, Wakayama-city, Wakayama 641-0012 (Japan); Akimoto, Shigeru [Department of Microbiology, Wakayama Medical University, 811-1 Kimiidera, Wakayama-city, Wakayama 641-0012 (Japan); Nomoto, Koji; Tanaka, Ryuichiro [Yakult Central Institute for Microbiological Research, Tokyo (Japan); Ichinose, Masao, E-mail: ichinose@wakayama-med.ac.jp [Department of Gastroenterology, Wakayama Medical University, 811-1 Kimiidera, Wakayama-city, Wakayama 641-0012 (Japan)

    2009-04-17

    To clarify the alterations in lower intestinal microflora induced by gastric acid reduction, the dynamics of 12 major genera or groups of bacteria comprising the microflora in feces and colonic contents were examined by quantitative real-time PCR in proton pump inhibitor-treated rats and in asymptomatic human subjects with hypochlorhydria. In both rat and human experiments, most genera or groups of intestinal microflora (facultative and obligate anaerobes) proliferated by gastric acid reduction, and marked and significant increases in the Lactobacilli group and Veillonella, oropharyngeal bacteria, were observed. In rats, potent gastric acid inhibition led to a marked and significant increase of intestinal bacteria, including the Bacteroidesfragilis group, while Bifidobacterium, a beneficial bacterial species, remained at a constant level. These results strongly indicate that the gastric acid barrier not only controls the colonization and growth of oropharyngeal bacteria, but also regulates the population and composition of lower intestinal microflora.

  2. Identification of midgut microvillar proteins from Tenebrio molitor and Spodoptera frugiperda by cDNA library screenings with antibodies.

    Science.gov (United States)

    Ferreira, A H P; Cristofoletti, P T; Lorenzini, D M; Guerra, L O; Paiva, P B; Briones, M R S; Terra, W R; Ferreira, C

    2007-11-01

    The objective of this study was to identify midgut microvillar proteins in insects appearing earlier (Coleoptera) and later (Lepidoptera) in evolution. For this, cytoskeleton-free midgut microvillar membrane from Spodoptera frugiperda (Lepidoptera) and Tenebrio molitor (Coleoptera) were used to raise antibodies. These were used for screening midgut cDNA expression libraries. Positive clones were sequenced, assembled and searched for similarities with gene/protein databases. The predicted midgut microvillar proteins from T. molitor were: cockroach allergens (unknown function), peritrophins (peritrophic membrane proteins), digestive enzymes (aminopeptidase, alpha-mannosidase) and unknown proteins. Predicted S. frugiperda midgut proteins may be grouped into six classes: (a) proteins involved in protection of midgut (thioredoxin peroxidase, aldehyde dehydrogenase, serpin and juvenile hormone epoxide hydrolase); (b) digestive enzymes (astacin, transporter-like amylase, aminopeptidase, and carboxypeptidase); (c) peritrophins; (d) proteins associated with microapocrine secretion (gelsolin, annexin); (e) membrane-tightly bound-cytoskeleton proteins (fimbrin, calmodulin) and (f) unidentified proteins. The novel approach is compared with others and microvillar function is discussed in the light of the predicted proteins.

  3. Gut microflora of vervet and samango monkeys in relation to diet.

    OpenAIRE

    Bruorton, M R; Davis, C. L.; Perrin, M R

    1991-01-01

    The microflora in the gastrointestinal tracts of wild vervet and samango monkeys (Cercopithecus aethiops and C. mitis, respectively) were studied, using fermentation acid analysis, electron microscopy, and culturing methods. The diets of the two species of monkey differ considerably, with that of the samango including a greater proportion of cellulose-rich leaf material, and this is reflected in the microflora. Volatile fatty acid measurements along the gut of both species showed that these e...

  4. [Recognition of commensal microflora by pattern recognition receptors in human physiology and pathology].

    Science.gov (United States)

    Bondarenko, V M; Likhoded, V G

    2012-01-01

    Contemporary data on the interaction of commensal microflora and Toll-like pattern recognition receptors are presented. These receptors recognize normal intestine microflora in physiological conditions, and this interaction is necessary for the maintenance of homeostasis and damage reparation of the intestine, for the induction of heat shock cytoprotective proteins. As a side effect in disruption of immunologic tolerance and misbalance of protective immunological mechanisms, multiorgan pathologic changes of organs and tissues may develop, including chronic inflammation processes of various localization.

  5. Effect of disturbance of the gastrointestinal microflora on the faecal excretion of Fusobacterium necrophorum biovar A.

    OpenAIRE

    Smith, G. R.; Thornton, E. A.

    1993-01-01

    Oral pretreatment of mice with either a mixture of kanamycin and erythromycin or metronidazole to modify the gut microflora greatly enhanced the faecal excretion of Fusobacterium necrophorum biovar A given by mouth. This lends support to the suggestion that disturbance of the gastrointestinal microflora in animals such as cattle, which often carry the organism in the rumen, may lead to intestinal multiplication and faecal excretion, thereby providing a source of infection that may lead to nec...

  6. Role of intestinal microflora in the degradation of DDT by rainbow trout (Salmo gairdneri)

    Science.gov (United States)

    Wedemeyer, G.

    1968-01-01

    Though liver homogenates show apparent microsomal enzyme DDT-dehydrochlorinase activity, in the intact fish the intestinal microflora play a major role in DDT detoxication. Since the presence of this microflora in fish depends on the recent intake of food (12), the rate of detoxication and hence the toxicity of ingested DDT to the rainbow trout will probably depend somewhat on the available food supply.

  7. Fine-structural changes in the midgut of old Drosophila melanogaster

    Science.gov (United States)

    Anton-Erxleben, F.; Miquel, J.; Philpott, D. E.

    1983-01-01

    Senescent fine-structural changes in the midgut of Drosophila melanogaster are investigated. A large number of midgut mitochondria in old flies exhibit nodular cristae and a tubular system located perpendicular to the normal cristae orientation. Anterior intestinal cells show a senescent accumulation of age pigment, either with a surrounding two-unit membrane or without any membrane. The predominant localization of enlarged mitochondria and pigment in the luminal gut region may be related to the polarized metabolism of the intestinal cells. Findings concur with previous observations of dense-body accumulations and support the theory that mitochondria are involved in the aging of fixed post-mitotic cells. Demonstrated by statistical analyses is that mitochondrial size increase is related to mitochondrial variation increase.

  8. Radiologic Findings of Reversed Intestinal Rotation in Adults: 3 Cases Report

    Energy Technology Data Exchange (ETDEWEB)

    Baek, Hyeon Seok; Cho, Jae Ho; Chang, Jay Chun; Kim, Jae Woon; Kim, Kum Rae [Yeungnam University, Gyeongsan (Korea, Republic of); Park, Won Kyu [Suh Joo Mir Radiologic Clinic, Daegu (Korea, Republic of); Kim, Jong Yeol [Gumi CHA University Medical Center, Gumi (Korea, Republic of)

    2009-12-15

    Most anomalies of intestinal rotation are detected during the postneonatal period. In adults, the diagnosis and treatment of patients with a congenital anomaly of the midgut can be difficult because of their extremely rarity. Based on embryology, anomalies of intestinal rotation can be divided into non-rotation, reversed rotation and malrotation. Reversed rotation of the midgut is the rarest of all anomalies of intestinal rotation. Although this anomaly is rare, it can be diagnosed by a detailed knowledge of embryology and anatomy. We report three adult patients with reversed intestinal rotation and review the embryology, clinical presentation and radiographic findings of this disorder

  9. Proteolytic activity in the midgut of Ectomyelois ceratoniae Zeller (Lepidoptera: Pyralidae), Pomegranate carob moth

    OpenAIRE

    M. Ranjbar; JJ Sendi; A Zibaee

    2011-01-01

    In this study, the proteolytic activity in the midgut of Ectomyelois ceratoniae as the major pest of pomegranate was investigated to find nature of specific proteases and their properties for adopting possible pest management procedure. It was found that fourth and fifth instar larvae had the highest proteolytic activity as well as specific proteinases including, elastase, trypsin-like, chymotrypsin-like and two exopeptidases. The optimal pH of general protease was 10 and 9 for azocasein, cas...

  10. Multiple Modes of Action of the Squamocin in the Midgut Cells of Aedes aegypti Larvae

    Science.gov (United States)

    de Paula, Sérgio Oliveira; Martins, Gustavo Ferreira; Zanuncio, José Cola

    2016-01-01

    Annonaceous acetogenins are botanical compounds with good potential for use as insecticides. In the vector, Aedes aegypti (L.) (Diptera: Culicidae), squamocin (acetogenin) has been reported to be a larvicide and cytotoxic, but the modes of action of this molecule are still poorly understood. This study evaluated the changes in the cell morphology, and in the expression of genes, for autophagy (Atg1 and Atg8), for membrane ion transporter V-ATPase, and for water channel aquaporin-4 (Aqp4) in the midgut of A. aegypti larvae exposed to squamocin from Annona mucosa Jacq. (Annonaceae). Squamocin showed cytotoxic action with changes in the midgut epithelium and digestive cells of A. aegypti larvae, increase in the expression for autophagy gene Atg1 and Atg8, decrease in the expression of V-ATPase, decrease in the expression of Aqp4 gene in LC20 and inhibition of Apq4 genes in the midgut of this vector in LC50. These multiple modes of action for squamocin are described for the first time in insects, and they are important because different sites of action of squamocin from A. mucosa may reduce the possibility of resistance of A. aegypti to this molecule. PMID:27532504

  11. Cytotoxic effects of thiamethoxam in the midgut and malpighian tubules of Africanized Apis mellifera (Hymenoptera: Apidae).

    Science.gov (United States)

    Catae, Aline Fernanda; Roat, Thaisa Cristina; De Oliveira, Regiane Alves; Nocelli, Roberta Cornélio Ferreira; Malaspina, Osmar

    2014-04-01

    Due to its expansion, agriculture has become increasingly dependent on the use of pesticides. However, the indiscriminate use of insecticides has had additional effects on the environment. These products have a broad spectrum of action, and therefore the insecticide affects not only the pests but also non-target insects such as bees, which are important pollinators of agricultural crops and natural environments. Among the most used pesticides, the neonicotinoids are particularly harmful. One of the neonicotinoids of specific concern is thiamethoxam, which is used on a wide variety of crops and is toxic to bees. Thus, this study aimed to analyze the effects of this insecticide in the midgut and Malpighian tubule cells of Africanized Apis mellifera. Newly emerged workers were exposed until 8 days to a diet containing a sublethal dose of thiamethoxam equal to 1/10 of LC₅₀ (0.0428 ng a.i./l L of diet). The bees were dissected and the organs were processed for transmission electron microscopy. The results showed that thiamethoxam is cytotoxic to midgut and Malpighian tubules. In the midgut, the damage was more evident in bees exposed to the insecticide on the first day. On the eighth day, the cells were ultrastructurally intact suggesting a recovery of this organ. The Malpighian tubules showed pronounced alterations on the eighth day of exposure of bees to the insecticide. This study demonstrates that the continuous exposure to a sublethal dose of thiamethoxam can impair organs that are used during the metabolism of the insecticide.

  12. Differential expression profiles in the midgut of Triatoma infestans infected with Trypanosoma cruzi.

    Directory of Open Access Journals (Sweden)

    Diego S Buarque

    Full Text Available Chagas disease, or American trypanosomiasis, is a parasitic disease caused by the protozoan Trypanosoma cruzi and is transmitted by insects from the Triatominae subfamily. To identify components involved in the protozoan-vector relationship, we constructed and analyzed cDNA libraries from RNA isolated from the midguts of uninfected and T. cruzi-infected Triatoma infestans, which are major vectors of Chagas disease. We generated approximately 440 high-quality Expressed Sequence Tags (ESTs from each T. infestans midgut cDNA library. The sequences were grouped in 380 clusters, representing an average length of 664.78 base pairs (bp. Many clusters were not classified functionally, representing unknown transcripts. Several transcripts involved in different processes (e.g., detoxification showed differential expression in response to T. cruzi infection. Lysozyme, cathepsin D, a nitrophorin-like protein and a putative 14 kDa protein were significantly upregulated upon infection, whereas thioredoxin reductase was downregulated. In addition, we identified several transcripts related to metabolic processes or immunity with unchanged expressions, including infestin, lipocalins and defensins. We also detected ESTs encoding juvenile hormone binding protein (JHBP, which seems to be involved in insect development and could be a target in control strategies for the vector. This work demonstrates differential gene expression upon T. cruzi infection in the midgut of T. infestans. These data expand the current knowledge regarding vector-parasite interactions for Chagas disease.

  13. Effects of dietary nickel on detoxification enzyme activities in the midgut of Spodoptera litura Fabricius larvae

    Institute of Scientific and Technical Information of China (English)

    SUN HongXia; ZHOU Qiang; TANG WenCheng; SHU YingHua; ZHANG GuRen

    2008-01-01

    Nickel accumulated in midugt of Spodoptera litura Fabricius could induce the expression of metal-Iothionein, one of the most important detoxification proteins in organisms. In the present study, the effects of dietary nickel on the activities of detoxification enzymes, such as carboxylesterase (CarE) and glutathione S-transferase (GST) in the midgut of S. litura larvae have been studied to get an un-derstanding of the detoxification mechanisms of S. litura larvae to excessive nickel. Results showed that CarE activities in the midgut of the 5th instar larvae decreased at lower levels of nickel (≤5 mg/kg), while increased with increasing nickel doses at higher levels of nickel (≥10 mg/kg) exposure in suc-cessive 3 generations. CarE activities of the 6th instar larvae were also characterized as inhibited at low levels of nickel exposure, and improved at higher levels in the 1st generation. CarE activities of 6th instar larvae in the 2rid and 3rd generations were all lower than that in control. However, GST activities in the midgut of the 5th and 6th instar larvae all increased with increasing nickel doses (1-20 mg/kg) in diets.

  14. Effects of tannic acid on trypsin and leucine aminopeptidase activities in gypsy moth larval midgut

    Directory of Open Access Journals (Sweden)

    Mrdaković Marija

    2013-01-01

    Full Text Available The effects of allelochemical stress on genetic variations in the specific activities of gypsy moth digestive enzymes (trypsin and leucine aminopeptidase and relative midgut mass (indirect measure of food consumption, as well as variability in their plasticity, were investigated in fifth instar gypsy moths originating from two populations with different trophic adaptations (oak and locust-tree forests. Thirty-two full-sib families from the Quercus population and twenty-six full-sib families from the Robinia population were reared on an artificial diet with or without supplementation with tannic acid. Between population differences were observed as higher average specific activity of trypsin and relative midgut mass in larvae from the Robinia population. Significant broad-sense heritabilities were observed for the specific activity of trypsin in the control state, and for specific activity of leucine aminopeptidase in a stressful environment. Significantly lower heritability for relative midgut mass was recorded in larvae from the Robinia population reared under stressful conditions. Significant variability of trypsin plasticity in larvae from both populations and significant variability of leucine aminopeptidase plasticity in larvae from the Robinia population point to the potential for the evolution of enzyme adaptive plastic responses to the presence of stressor. Non-significant across-environment genetic correlations do not represent a constraint for the evolution of enzyme plasticity. [Projekat Ministarstva nauke Republike Srbije, br. 173027

  15. High quality RNA isolation from Aedes aegypti midguts using laser microdissection microscopy

    Directory of Open Access Journals (Sweden)

    Gobert Geoffrey N

    2011-05-01

    Full Text Available Abstract Background Laser microdissection microscopy (LMM has potential as a research tool because it allows precise excision of target tissues or cells from a complex biological specimen, and facilitates tissue-specific sample preparation. However, this method has not been used in mosquito vectors to date. To this end, we have developed an LMM method to isolate midgut RNA using Aedes aegypti. Results Total RNA was isolated from Ae. aegypti midguts that were either fresh-frozen or fixed with histological fixatives. Generally, fresh-frozen tissue sections are a common source of quality LMM-derived RNA; however, our aim was to develop an LMM protocol that could inactivate pathogenic viruses by fixation, while simultaneously preserving RNA from arbovirus-infected mosquitoes. Three groups (10 - 15 mosquitoes per group of female Ae. aegypti at 24 or 48-hours post-blood meal were intrathoracically injected with one of seven common fixatives (Bouin's, Carnoy's, Formoy's, Cal-Rite, 4% formalin, 10% neutral buffered formalin, or zinc formalin to evaluate their effect on RNA quality. Total RNA was isolated from the fixed abdomens using a Trizol® method. The results indicated that RNA from Carnoy's and Bouin's fixative samples was comparable to that of fresh frozen midguts (control in duplicate experiments. When Carnoy's and Bouin's were used to fix the midguts for the LMM procedure, however, Carnoy's-fixed RNA clearly showed much less degradation than Bouin's-fixed RNA. In addition, a sample of 5 randomly chosen transcripts were amplified more efficiently using the Carnoy's treated LMM RNA than Bouin's-fixed RNA in quantitative real-time PCR (qRT-PCR assays, suggesting there were more intact target mRNAs in the Carnoy's fixed RNA. The yields of total RNA ranged from 0.3 to 19.0 ng per ~3.0 × 106 μm2 in the LMM procedure. Conclusions Carnoy's fixative was found to be highly compatible with LMM, producing high quality RNA from Ae. aegypti midguts while

  16. A lepidopteran-specific gene family encoding valine-rich midgut proteins.

    Directory of Open Access Journals (Sweden)

    Jothini Odman-Naresh

    Full Text Available Many lepidopteran larvae are serious agricultural pests due to their feeding activity. Digestion of the plant diet occurs mainly in the midgut and is facilitated by the peritrophic matrix (PM, an extracellular sac-like structure, which lines the midgut epithelium and creates different digestive compartments. The PM is attracting increasing attention to control lepidopteran pests by interfering with this vital function. To identify novel PM components and thus potential targets for insecticides, we performed an immunoscreening with anti-PM antibodies using an expression library representing the larval midgut transcriptome of the tobacco hornworm, Manduca sexta. We identified three cDNAs encoding valine-rich midgut proteins of M. sexta (MsVmps, which appear to be loosely associated with the PM. They are members of a lepidopteran-specific family of nine VMP genes, which are exclusively expressed in larval stages in M. sexta. Most of the MsVMP transcripts are detected in the posterior midgut, with the highest levels observed for MsVMP1. To obtain further insight into Vmp function, we expressed MsVMP1 in insect cells and purified the recombinant protein. Lectin staining and glycosidase treatment indicated that MsVmp1 is highly O-glycosylated. In line with results from qPCR, immunoblots revealed that MsVmp1 amounts are highest in feeding larvae, while MsVmp1 is undetectable in starving and molting larvae. Finally using immunocytochemistry, we demonstrated that MsVmp1 localizes to the cytosol of columnar cells, which secrete MsVmp1 into the ectoperitrophic space in feeding larvae. In starving and molting larvae, MsVmp1 is found in the gut lumen, suggesting that the PM has increased its permeability. The present study demonstrates that lepidopteran species including many agricultural pests have evolved a set of unique proteins that are not found in any other taxon and thus may reflect an important adaptation in the highly specialized lepidopteran

  17. A lepidopteran-specific gene family encoding valine-rich midgut proteins.

    Science.gov (United States)

    Odman-Naresh, Jothini; Duevel, Margret; Muthukrishnan, Subbaratnam; Merzendorfer, Hans

    2013-01-01

    Many lepidopteran larvae are serious agricultural pests due to their feeding activity. Digestion of the plant diet occurs mainly in the midgut and is facilitated by the peritrophic matrix (PM), an extracellular sac-like structure, which lines the midgut epithelium and creates different digestive compartments. The PM is attracting increasing attention to control lepidopteran pests by interfering with this vital function. To identify novel PM components and thus potential targets for insecticides, we performed an immunoscreening with anti-PM antibodies using an expression library representing the larval midgut transcriptome of the tobacco hornworm, Manduca sexta. We identified three cDNAs encoding valine-rich midgut proteins of M. sexta (MsVmps), which appear to be loosely associated with the PM. They are members of a lepidopteran-specific family of nine VMP genes, which are exclusively expressed in larval stages in M. sexta. Most of the MsVMP transcripts are detected in the posterior midgut, with the highest levels observed for MsVMP1. To obtain further insight into Vmp function, we expressed MsVMP1 in insect cells and purified the recombinant protein. Lectin staining and glycosidase treatment indicated that MsVmp1 is highly O-glycosylated. In line with results from qPCR, immunoblots revealed that MsVmp1 amounts are highest in feeding larvae, while MsVmp1 is undetectable in starving and molting larvae. Finally using immunocytochemistry, we demonstrated that MsVmp1 localizes to the cytosol of columnar cells, which secrete MsVmp1 into the ectoperitrophic space in feeding larvae. In starving and molting larvae, MsVmp1 is found in the gut lumen, suggesting that the PM has increased its permeability. The present study demonstrates that lepidopteran species including many agricultural pests have evolved a set of unique proteins that are not found in any other taxon and thus may reflect an important adaptation in the highly specialized lepidopteran digestive tract facing

  18. Impact of Bifidobacterium longum and Lactobacillus acidophilus on the intestinal microflora and bioavailability of some food mutagens

    OpenAIRE

    Orrhage, Kerstin

    1999-01-01

    The gastrointestinal microflora is a complex ecological system, normally characterised by a flexible equilibrium. The most important role of the microflora, from the point of view of the host, is probably to act in colonisation resistance against exogenous, potentially pathogenic, microorganisms. The microflora also performs an important metabolic activity, breaking down and transforming compounds, in the intestine. Bifidobacteria and lactobacilli are Gram positive lactic ac...

  19. Blood meal-derived heme decreases ROS levels in the midgut of Aedes aegypti and allows proliferation of intestinal microbiota.

    Directory of Open Access Journals (Sweden)

    Jose Henrique M Oliveira

    2011-03-01

    Full Text Available The presence of bacteria in the midgut of mosquitoes antagonizes infectious agents, such as Dengue and Plasmodium, acting as a negative factor in the vectorial competence of the mosquito. Therefore, knowledge of the molecular mechanisms involved in the control of midgut microbiota could help in the development of new tools to reduce transmission. We hypothesized that toxic reactive oxygen species (ROS generated by epithelial cells control bacterial growth in the midgut of Aedes aegypti, the vector of Yellow fever and Dengue viruses. We show that ROS are continuously present in the midgut of sugar-fed (SF mosquitoes and a blood-meal immediately decreased ROS through a mechanism involving heme-mediated activation of PKC. This event occurred in parallel with an expansion of gut bacteria. Treatment of sugar-fed mosquitoes with increased concentrations of heme led to a dose dependent decrease in ROS levels and a consequent increase in midgut endogenous bacteria. In addition, gene silencing of dual oxidase (Duox reduced ROS levels and also increased gut flora. Using a model of bacterial oral infection in the gut, we show that the absence of ROS resulted in decreased mosquito resistance to infection, increased midgut epithelial damage, transcriptional modulation of immune-related genes and mortality. As heme is a pro-oxidant molecule released in large amounts upon hemoglobin degradation, oxidative killing of bacteria in the gut would represent a burden to the insect, thereby creating an extra oxidative challenge to the mosquito. We propose that a controlled decrease in ROS levels in the midgut of Aedes aegypti is an adaptation to compensate for the ingestion of heme.

  20. Fermentation and epiphytic microflora dynamics in sugar cane silage

    Directory of Open Access Journals (Sweden)

    Pedroso André de Faria

    2005-01-01

    Full Text Available Sugar cane silages are characterized by extensive yeast activity, alcohol production and great dry matter - DM - losses. Better knowledge of the fermentation process is fundamental to the development of efficient ensilage techniques for this forage. This study evaluates temporal changes in chemical composition, DM losses and epiphytic microflora in sugar cane silage. Mature sugar cane, variety RB835486 (12 months of vegetative growth, was hand harvested, processed in a stationary chopper and ensiled in 20-L plastic buckets provided with valves for gas release and a device for effluent collection. Laboratory silos were kept at ambient temperature and sampled after fraction one-half, 1, 2, 3, 7, 15, 45, 90, 120 and 180 days. Ethanol concentration reached 6.4% in DM after 15 days of ensilage, followed by 71% water soluble carbohydrates - WSCs - disappearance. Gas and total DM losses reached a plateau on day 45 (16% and 29% of DM, respectively. Yeast count was higher on the second day (5.05 log cfu g-1. Silage pH declined to below 4.0 on the third day. Effluent yield was negligible (20 kg t-1. DM content in the forage decreased (35% to 26% from day 0 to day 45. The increase in ethanol concentration showed an opposite trend to WSCs and true in vitro dry matter digestibility reductions in the silage. Developing methods to control yeasts, most probably through the use of additives, will enable more efficient production of sugar cane silage by farmers.

  1. Effect of leavening microflora on pizza dough properties.

    Science.gov (United States)

    Coppola, S; Pepe, O; Mauriello, G

    1998-11-01

    Fourteen different starter cultures containing one strain of Saccharomyces cerevisiae with and without individual or combinations of lactic acid bacteria (Lactobacillus plantarum, Lact. sanfrancisco, Enterococcus faecium, Leuconostoc mesenteroides) were employed to investigate the role of leavening microflora on the properties of pizza doughs. Microbiological, chemical and physical characteristics of doughs prepared with the same flour and under the same processing conditions were determined. Leavening times and acidification properties depended on the microbial association used. The proportions of lactic and acetic acid produced by lactic acid bacteria were consistent with the metabolic properties of the strains employed. The bacteria/yeast ratios arising from microbial counts at the end of the leavening process were always lower in comparison to sour- or bread-doughs. The size of the yeast population did not change much, while bacteria showed from one to four duplications. Rheologically, the fermented doughs could only be significantly distinguished from the control dough with regard to the elastic modulus. Principal Component Analysis was applied to the acidimetric data. The scattergram of the two principal components effectively discriminated 13 of the 14 pizza dough types.

  2. Bioconversion of corncob to hydrogen using anaerobic mixed microflora

    Energy Technology Data Exchange (ETDEWEB)

    Pan, Chunmei [Department of Chemistry, Zhengzhou University, Daxue Road, Zhengzhou 450052 (China); Biotechnology Department, Zhengzhou College of Animal Husbandry Engineering, Zhengzhou 450011 (China); Zhang, Shufang; Fan, Yaoting; Hou, Hongwei [Department of Chemistry, Zhengzhou University, Daxue Road, Zhengzhou 450052 (China)

    2010-04-15

    Biohydrogen production from corncob using natural anaerobic microflora was reported for the first time. The optimum pretreatment condition for the corncob was determined to be 100 C, 30 min, and 1% HCl (w/w). The maximum hydrogen yield of 107.9 ml/g-TVS and hydrogen production rate of 4.20 ml/g-TVS h{sup -1} was obtained under the condition of 10 g/l substrate concentration and initial pH 8.0. Butyrate and acetate were the dominant metabolic by-products of hydrogen fermentation. Chemical composition analysis, Fourier transform infrared spectroscopy (FTIR) and X-ray diffraction (XRD) were used to study the mechanism of degrading corncob for hydrogen production. The amorphous domains of cellulose and hemicellulose were hydrolyzed into fermentable saccharides through acid pretreatment and the microorganisms had a devastating effect on the crystallinity of the cellulose. The hydrogen yield from pretreated corncob was much higher than from raw corncob. Therefore, the acid pretreatment played a crucial role on hydrogen production from corncob. (author)

  3. Microbiological toxicity of tilmicosin on human colonic microflora in chemostats.

    Science.gov (United States)

    Hao, Haihong; Yao, Junping; Wu, Qinghua; Wei, Yajing; Dai, Menghong; Iqbal, Zahid; Wang, Xu; Wang, Yulian; Huang, Lingli; Chen, Dongmei; Tao, Yanfei; Liu, Zhenli; Yuan, Zonghui

    2015-10-01

    To evaluate the microbiological safety of tilmicosin on human intestinal microflora, four chemostat models of healthy human colonic ecosystems were exposed to tilmicosin (0, 0.436, 4.36, and 43.6 μg/mL) for 7 days. Prior to and during drug exposure, three microbiological endpoints were monitored daily including short-chain fatty acids, bacterial counts and macrolide susceptibility. Colonization resistance of each community was determined by 3 successive daily challenges of Salmonella typhimurium. Genes associated with virulence and macrolide resistance in Enterococcus faecalis were determined by PCR. Transcriptional expression of the virulence gene (gelE) in E. faecalis was determined by real-time RT-PCR. Our results showed that different concentrations of tilmicosin did not disrupt the colonization resistance in each chemostat. During exposure to 4.36 and 43.6 μg/mL tilmicosin, the Bacteroides fragilis population was significantly decreased while the proportion of resistant Enterococci increased. After long-term exposure to the highest concentration (43.6 μg/mL) of tilmicosin, the gelE gene was significantly up-regulated in the high-level macrolide resistant strains that also contained the ermB resistance gene. This study was the first of its kind to evaluate the microbiological toxicity of tilmicosin using a chemostat model. These findings also provide new insight into the co-occurrence of macrolide resistance and virulence in E. faecalis under tilmicosin selective pressure.

  4. Response of the common cutworm Spodoptera litura to zinc stress: Zn accumulation, metallothionein and cell ultrastructure of the midgut

    Energy Technology Data Exchange (ETDEWEB)

    Shu, Yinghua [Key Laboratory of Agro-Environments in Tropics, Ministry of Agriculture, South China Agricultural University, Guangzhou 510642 (China); Key Laboratory of Agroecology and Rural Environment of Guangdong Regular Higher Education Institutions, South China Agricultural University, Guangzhou 510642 (China); Institute of Tropical and Subtropical Ecology, South China Agricultural University, Guangzhou 510642 (China); State Key Laboratory of Biological Control and Institute of Entomology, Sun Yat-sen University, Guangzhou 510275 (China); Zhang, Guren [State Key Laboratory of Biological Control and Institute of Entomology, Sun Yat-sen University, Guangzhou 510275 (China); Wang, Jianwu, E-mail: wangjw@scau.edu.cn [Key Laboratory of Agro-Environments in Tropics, Ministry of Agriculture, South China Agricultural University, Guangzhou 510642 (China); Key Laboratory of Agroecology and Rural Environment of Guangdong Regular Higher Education Institutions, South China Agricultural University, Guangzhou 510642 (China); Institute of Tropical and Subtropical Ecology, South China Agricultural University, Guangzhou 510642 (China)

    2012-11-01

    By exposing the common cutworm Spodoptera litura Fabricius larvae to a range of Zinc (Zn) stress, we investigated the effects of dietary Zn on Zn accumulation, metallothionein (MT), and on the ultrastructure of the midgut. The techniques we used were inductively coupled plasma-atomic emission spectrometer (ICP-AES), real-time PCR combined with cadmium-hemoglobin total saturation, and transmission electron microscopy (TEM), respectively. There was a significant dose-response relationship between the Zn accumulations in the midgut of the larvae and the Zn concentrations in the diet. Furthermore, both MT content and MT gene expression in the midgut were significantly induced in the 50-500 mg Zn/kg treatments, and were significantly positively correlated with the Zn accumulations in the midgut. When S. litura larvae were fed with the diet treated with 500 mg Zn/kg, Zn accumulation and MT content in the midgut was 4450.85 mg Zn/kg and 372.77 mg/kg, respectively, thereafter there was a little increase; the level of MT gene expression was maximal, thereafter there was a sharp decrease. TEM showed that numerous electron-dense granules (EDGs) and vacuoles appeared in the cytoplasm of the midgut cells, their number and size being closely correlated with the Zn accumulations in the midgut. Moreover, the nuclei were strongly influenced by Zn stress, evidenced by chromatin condensation and irregular nuclear membranes. Therefore, after being exposed to Zn in the threshold (500 mg Zn/kg) range, S. litura larvae could accumulate Zn in the midgut, which led to the induction of MT and changes in cell ultrastructure (mainly the presence of EDGs). The induction of MT and precipitation of Zn in EDGs may be the effective detoxification mechanisms by which the herbivorous insect S. litura defends itself against heavy metals. -- Graphical abstract: When the herbivorous insect Spodoptera litura Fabricius larvae were fed on the artificial diet with different concentrations of Zn, amounts of

  5. Metallothionein expression induced by nickel accumulation in the midgut of Spodoptera litura Fabricius larvae exposed to nickel

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    Metallothionein expression induced by nickel accumulation in the midgut of Spodoptera litura Fabricius larvae was investigated by exposing S. litura larvae to the artificial diets amended with different doses of nickel for 3 generations. Nickel accumulations in the midgut of 6th instar larvae were detected by Inductively Coupled Plasma-Atomic Emission Spectrometer (ICP-AES), and the induced metallothionein expression in the midgut of 5th and 6th instar larvae in 120 h were quantified using cadmium-hemoglobin total saturation methods. The accumulated nickel in the midgut of the 6th instar larvae within a generation increased with the increase of the nickel doses in the treated diets and showed significant dose-dependence with the nickel doses in diets. Metallothioneins induced by nickel accumulated in the larval midgut also increased with nickel doses in diets and with the increasing stressing time in a generation. Results also suggested that the metallothionein expression was different with the time of S. litura larvae stressed by nickel.

  6. [Oral cavity microflora in patients with non-specific ulcerative colitis and Crohn's disease].

    Science.gov (United States)

    Kondrakova, O A; Muliar, E A; Voropaeva, E A; Babin, V N; Dubinin, A V; Briko, N I

    2009-01-01

    Results of study of microecological disorders in oral cavity of patients with non-specific ulcerative colitis (NSUC) and Crohn's disease (CD) and control subjects (patients with hypertension). Condition of mucosa was assessed on the basis of morphological data and electrophoretic mobility of cell nuclei, whereas structure of microbiocenosis and metabolic activity of microflora--on the basis of saliva bacterial culture and contents and profile of volatile fat acids in it. Detection rate of negative charge of the cell nuclei (decrease of functional activity of epithelium) was significantly higher in patients with NSUC and CD (66.6%) compared with controls (10%). This fact was directly related with hypercolonization of oral cavity by Gram-negative microflora. Lesions of mucosa which are characteristic of NSUC and CD and determined by pathologic immune mechanisms correlated with quantity of pathogenic microflora (Staphylococcus aureus and Candida). Marked differences of chromatograms' patterns were observed in patients with NSUC and CD indicating the suppression of anaerobic microflora in patients with CD and hypercolonization of oral cavity by anaerobic microflora in majority of patients with NSUC.

  7. Midgut-enriched receptor protein tyrosine phosphatase PTP52F is required for Drosophila development during larva-pupa transition.

    Science.gov (United States)

    Santhanam, Abirami; Liang, Suh-Yuen; Chen, Dong-Yuan; Chen, Guang-Chao; Meng, Tzu-Ching

    2013-01-01

    To date our understanding of Drosophila receptor protein tyrosine phosphatases (R-PTPs) in the regulation of signal transduction is limited. Of the seven R-PTPs identified in flies, six are involved in the axon guidance that occurs during embryogenesis. However, whether and how R-PTPs may control key steps of Drosophila development is not clear. In this study we investigated the potential role of Drosophila R-PTPs in developmental processes outside the neuronal system and beyond the embryogenesis stage. Through systematic data mining of available microarray databases, we found the mRNA level of PTP52F to be highly enriched in the midgut of flies at the larva-pupa transition. This finding was confirmed by gut tissue staining with a specific antibody. The unique spatiotemporal expression of PTP52F suggests that it is possibly involved in regulating metamorphosis during the transformation from larva to pupa. To test this hypothesis, we employed RNA interference to examine the defects of transgenic flies. We found that ablation of endogenous PTP52F led to high lethality characterized by the pharate adult phenotype, occurring due to post pupal eclosion failure. These results show that PTP52F plays an indispensable role during the larva-pupa transition. We also found that PTP52F could be reclassified as a member of the subtype R3 PTPs instead of as an unclassified R-PTP without a human ortholog, as suggested previously. Together, these findings suggest that Drosophila R-PTPs may control metamorphosis and other biological processes beyond our current knowledge.

  8. MDL28170, a calpain inhibitor, affects Trypanosoma cruzi metacyclogenesis, ultrastructure and attachment to Rhodnius prolixus midgut.

    Directory of Open Access Journals (Sweden)

    Vítor Ennes-Vidal

    Full Text Available BACKGROUND: Trypanosoma cruzi is the etiological agent of Chagas' disease. During the parasite life cycle, many molecules are involved in the differentiation process and infectivity. Peptidases are relevant for crucial steps of T. cruzi life cycle; as such, it is conceivable that they may participate in the metacyclogenesis and interaction with the invertebrate host. METHODOLOGY/PRINCIPAL FINDINGS: In this paper, we have investigated the effect of the calpain inhibitor MDL28170 on the attachment of T. cruzi epimastigotes to the luminal midgut surface of Rhodnius prolixus, as well as on the metacyclogenesis process and ultrastructure. MDL28170 treatment was capable of significantly reducing the number of bound epimastigotes to the luminal surface midgut of the insect. Once the cross-reactivity of the anti-Dm-calpain was assessed, it was possible to block calpain molecules by the antibody, leading to a significant reduction in the capacity of adhesion to the insect guts by T. cruzi. However, the antibodies were unable to interfere in metacyclogenesis, which was impaired by the calpain inhibitor presenting a significant reduction in the number of metacyclic trypomastigotes. The calpain inhibitor also promoted a direct effect against bloodstream trypomastigotes. Ultrastructural analysis of epimastigotes treated with the calpain inhibitor revealed disorganization in the reservosomes, Golgi and plasma membrane disruption. CONCLUSIONS/SIGNIFICANCE: The presence of calpain and calpain-like molecules in a wide range of organisms suggests that these proteins could be necessary for basic cellular functions. Herein, we demonstrated the effects of MDL28170 in crucial steps of the T. cruzi life cycle, such as attachment to the insect midgut and metacyclogenesis, as well as in parasite viability and morphology. Together with our previous findings, these results help to shed some light on the functions of T. cruzi calpains. Considering the potential roles of

  9. Restriction of Francisella novicida genetic diversity during infection of the vector midgut.

    Directory of Open Access Journals (Sweden)

    Kathryn E Reif

    2014-10-01

    Full Text Available The genetic diversity of pathogens, and interactions between genotypes, can strongly influence pathogen phenotypes such as transmissibility and virulence. For vector-borne pathogens, both mammalian hosts and arthropod vectors may limit pathogen genotypic diversity (number of unique genotypes circulating in an area by preventing infection or transmission of particular genotypes. Mammalian hosts often act as "ecological filters" for pathogen diversity, where novel variants are frequently eliminated because of stochastic events or fitness costs. However, whether vectors can serve a similar role in limiting pathogen diversity is less clear. Here we show using Francisella novicida and a natural tick vector of Francisella spp. (Dermacentor andersoni, that the tick vector acted as a stronger ecological filter for pathogen diversity compared to the mammalian host. When both mice and ticks were exposed to mixtures of F. novicida genotypes, significantly fewer genotypes co-colonized ticks compared to mice. In both ticks and mice, increased genotypic diversity negatively affected the recovery of available genotypes. Competition among genotypes contributed to the reduction of diversity during infection of the tick midgut, as genotypes not recovered from tick midguts during mixed genotype infections were recovered from tick midguts during individual genotype infection. Mediated by stochastic and selective forces, pathogen genotype diversity was markedly reduced in the tick. We incorporated our experimental results into a model to demonstrate how vector population dynamics, especially vector-to-host ratio, strongly affected pathogen genotypic diversity in a population over time. Understanding pathogen genotypic population dynamics will aid in identification of the variables that most strongly affect pathogen transmission and disease ecology.

  10. Tigutcystatin, a cysteine protease inhibitor from Triatoma infestans midgut expressed in response to Trypanosoma cruzi

    Energy Technology Data Exchange (ETDEWEB)

    Buarque, Diego S.; Spindola, Leticia M.N. [Department of Biochemistry, Universidade Federal de Sao Paulo, Escola Paulista de Medicina, 04044-020 Sao Paulo, SP (Brazil); Martins, Rafael M. [Biology of Host Parasite Interactions Unit, Institute Pasteur, 75015 Paris (France); Braz, Gloria R.C. [Department of Biochemistry, Instituto de Quimica, Universidade Federal do Rio de Janeiro, 21941-909 Rio de Janeiro (Brazil); Tanaka, Aparecida S., E-mail: Tanaka.bioq@epm.br [Department of Biochemistry, Universidade Federal de Sao Paulo, Escola Paulista de Medicina, 04044-020 Sao Paulo, SP (Brazil)

    2011-09-23

    Highlights: {yields} Tigutcystatin inhibits Trypanosoma cruzi cysteine proteases with high specificity. {yields} Tigutcystatin expression is up-regulated in response to T. cruzi infection. {yields} It is the first cysteine proteases inhibitor characterized from a triatomine insect. -- Abstract: The insect Triatoma infestans is a vector of Trypanosoma cruzi, the etiological agent of Chagas disease. A cDNA library was constructed from T. infestans anterior midgut, and 244 clones were sequenced. Among the EST sequences, an open reading frame (ORF) with homology to a cystatin type 2 precursor was identified. Then, a 288-bp cDNA fragment encoding mature cystatin (lacking signal peptide) named Tigutcystatin was cloned fused to a N-terminal His tag in pET-14b vector, and the protein expressed in Escherichia coli strain Rosetta gami. Tigutcystatin purified and cleaved by thrombin to remove His tag presented molecular mass of 11 kDa and 10,137 Da by SDS-PAGE and MALDI-TOF mass spectrometry, respectively. Purified Tigutcystatin was shown to be a tight inhibitor towards cruzain, a T. cruzi cathepsin L-like enzyme (K{sub i} = 3.29 nM) and human cathepsin L (K{sub i} = 3.78 nM). Tissue specific expression analysis showed that Tigutcystatin was mostly expressed in anterior midgut, although amplification in small intestine was also detected by semi quantitative RT-PCR. qReal time PCR confirmed that Tigutcystatin mRNA is significantly up-regulated in anterior midgut when T. infestans is infected with T. cruzi. Together, these results indicate that Tigutcystatin may be involved in modulation of T. cruzi in intestinal tract by inhibiting parasite cysteine proteases, which represent the virulence factors of this protozoan.

  11. Tissue-Specific Transcriptome Profiling of Plutella Xylostella Third Instar Larval Midgut

    Directory of Open Access Journals (Sweden)

    Wen Xie, Yanyuan Lei, Wei Fu, Zhongxia Yang, Xun Zhu, Zhaojiang Guo, Qingjun Wu, Shaoli Wang, Baoyun Xu, Xuguo Zhou, Youjun Zhang

    2012-01-01

    Full Text Available The larval midgut of diamondback moth, Plutella xylostella, is a dynamic tissue that interfaces with a diverse array of physiological and toxicological processes, including nutrient digestion and allocation, xenobiotic detoxification, innate and adaptive immune response, and pathogen defense. Despite its enormous agricultural importance, the genomic resources for P. xylostella are surprisingly scarce. In this study, a Bt resistant P. xylostella strain was subjected to the in-depth transcriptome analysis to identify genes and gene networks putatively involved in various physiological and toxicological processes in the P. xylostella larval midgut.Using Illumina deep sequencing, we obtained roughly 40 million reads containing approximately 3.6 gigabases of sequence data. De novo assembly generated 63,312 ESTs with an average read length of 416bp, and approximately half of the P. xylostella sequences (45.4%, 28,768 showed similarity to the non-redundant database in GenBank with a cut-off E-value below 10-5. Among them, 11,092 unigenes were assigned to one or multiple GO terms and 16,732 unigenes were assigned to 226 specific pathways. In-depth analysis indentified genes putatively involved in insecticide resistance, nutrient digestion, and innate immune defense. Besides conventional detoxification enzymes and insecticide targets, novel genes, including 28 chymotrypsins and 53 ABC transporters, have been uncovered in the P. xylostella larval midgut transcriptome; which are potentially linked to the Bt toxicity and resistance. Furthermore, an unexpectedly high number of ESTs, including 46 serpins and 7 lysozymes, were predicted to be involved in the immune defense.As the first tissue-specific transcriptome analysis of P. xylostella, this study sheds light on the molecular understanding of insecticide resistance, especially Bt resistance in an agriculturally important insect pest, and lays the foundation for future functional genomics research. In

  12. Biogeographical note on Antarctic microflorae: Endemism and cosmopolitanism

    Directory of Open Access Journals (Sweden)

    Waqar Azeem Jadoon

    2013-11-01

    Full Text Available This study deals with the biogeography of Antarctic microflora (Antarctica acts as best model to study microbial biogeography such as cyanobacteria and selected halophiles with special emphasis on Halomonas variabilis and Bacillus licheniformis. Halophiles are known to be resistant not only to salt stress, but also to extreme temperature, pressure, and aridity and they are capable of surviving in harsh environments such as polar regions, deep-sea habitats, and deserts. Many microbes are known to be resistant to hostile environmental conditions, and are capable of surviving in harsh environments. Our group has isolated 444 strains belonging to 28 genera of halophiles from various environments around the world. The 16S rRNA gene sequences revealed that many of the isolated strains from geographically distant habitats having different environmental conditions, were closely related to each other, with some strains possessing 100% identical sequences. Organisms possessing survival mechanism such as spore formation are usually ubiquitous. The genus Halomonas is represented by potentially endemic strains and the ubiquitous H. variabilis, while spore-forming B. licheniformis showed cosmopolitan distribution. One potentially endemic (moderate endemicity that is regional and/or continental distribution strain was reported from Syowa station, East Antarctica, and Mario Zucchelli station, West Antarctica, which are geographically separated by 3000 km. Moreover, 15 strains having 100% similarity with B. licheniformis were considered cosmopolitans. The results of this work provide support for the middle-ground model that some microbes have moderate endemicity and others have cosmopolitan distribution. These results will contribute to a greater understanding of microbial biogeography with special emphasis on Antarctica.

  13. A small molecule glycosaminoglycan mimetic blocks Plasmodium invasion of the mosquito midgut.

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    Derrick K Mathias

    Full Text Available Malaria transmission-blocking (T-B interventions are essential for malaria elimination. Small molecules that inhibit the Plasmodium ookinete-to-oocyst transition in the midgut of Anopheles mosquitoes, thereby blocking sporogony, represent one approach to achieving this goal. Chondroitin sulfate glycosaminoglycans (CS-GAGs on the Anopheles gambiae midgut surface are putative ligands for Plasmodium falciparum ookinetes. We hypothesized that our synthetic polysulfonated polymer, VS1, acting as a decoy molecular mimetic of midgut CS-GAGs confers malaria T-B activity. In our study, VS1 repeatedly reduced midgut oocyst development by as much as 99% (P<0.0001 in mosquitoes fed with P. falciparum and Plasmodium berghei. Through direct-binding assays, we observed that VS1 bound to two critical ookinete micronemal proteins, each containing at least one von Willebrand factor A (vWA domain: (i circumsporozoite protein and thrombospondin-related anonymous protein-related protein (CTRP and (ii vWA domain-related protein (WARP. By immunofluorescence microscopy, we observed that VS1 stains permeabilized P. falciparum and P. berghei ookinetes but does not stain P. berghei CTRP knockouts or transgenic parasites lacking the vWA domains of CTRP while retaining the thrombospondin repeat region. We produced structural homology models of the first vWA domain of CTRP and identified, as expected, putative GAG-binding sites on CTRP that align closely with those predicted for the human vWA A1 domain and the Toxoplasma gondii MIC2 adhesin. Importantly, the models also identified patches of electropositive residues that may extend CTRP's GAG-binding motif and thus potentiate VS1 binding. Our molecule binds to a critical, conserved ookinete protein, CTRP, and exhibits potent malaria T-B activity. This study lays the framework for a high-throughput screen of existing libraries of safe compounds to identify those with potent T-B activity. We envision that such compounds when

  14. Protein expression in the midgut of sugar-fed Aedes albopictus females

    Directory of Open Access Journals (Sweden)

    Saboia-Vahia Leonardo

    2012-12-01

    Full Text Available Abstract Background Aedes albopictus is a vector for several fatal arboviruses in tropical and sub-tropical regions of the world. The midgut of the mosquito is the first barrier that pathogens must overcome to establish infection and represents one of the main immunologically active sites of the insect. Nevertheless, little is known about the proteins involved in the defense against pathogens, and even in the processing of food, and the detoxification of metabolites. The identification of proteins exclusively expressed in the midgut is the first step in understanding the complex physiology of this tissue and can provide insight into the mechanisms of pathogen-vector interaction. However, identification of the locally expressed proteins presents a challenge because the Ae. albopictus genome has not been sequenced. Methods In this study, two-dimensional electrophoresis (2DE was combined with liquid chromatography in line with tandem mass spectrometry (LC-MS/MS and data mining to identify the major proteins in the midgut of sugar-fed Ae. albopictus females. Results Fifty-six proteins were identified by sequence similarity to entries from the Ae. aegypti genome. In addition, two hypothetical proteins were experimentally confirmed. According to the gene ontology analysis, the identified proteins were classified into 16 clusters of biological processes. Use of the STRING database to investigate protein functional associations revealed five functional networks among the identified proteins, including a network for carbohydrate and amino acid metabolism, a group associated with ATP production and a network of proteins that interact during detoxification of toxic free radicals, among others. This analysis allowed the assignment of a potential role for proteins with unknown function based on their functional association with other characterized proteins. Conclusion Our findings represent the first proteome map of the Ae. albopictus midgut and denotes the

  15. Antagonistic activity of autosimbionts А. viridans, B. subtilis and their probiotic association to conditionally microflora

    Directory of Open Access Journals (Sweden)

    Stepansky D.A.

    2015-03-01

    Full Text Available In this research the data on examination of antagonist qualities of bioassotiantes A. viridans and strain B. subtilis 3 towards pathogenic and opportunistic pathogenic microflora isolated from oropharynx and nasopharynx of children who were in contact with patients with pulmonary tuberculosis (MBT + are submitted. The expressed antagonist activity of autosimbionts A. viridans towards pathogenic and opportunistic pathogenic microflora was shown. Common antagonist activity of A. viridans (k N 1 and B. subtilis 3 towards diverse strains of test-cultures is 1,5-2 times higher, than separate antagonist activity of A. viridans (k №1 and B. subtilis 3. Received research data showed the possibility of continuing work on development of probiotic associations, that contain representatives of normal microflora - bioassociants A. viridans and probiotic strains B. subtilis 3 with broadspectrum of antagonistic activity in relation to the various groups of bacterium.

  16. The Interaction of Large Bowel Microflora with the Colonic Mucus Barrier

    Directory of Open Access Journals (Sweden)

    Jeffrey P. Pearson

    2010-01-01

    Full Text Available The colonic mucus barrier is the first line of defence that the underlying mucosa has against the wide range of potentially damaging agents of microbial, endogenous, and dietary origin that occur within the colonic lumen. The functional component of mucus is the secreted, polymeric glycoprotein mucin. The mucus barrier can either act as an energy source or a support medium for growth to the intestinal microflora. The mucus barrier appears to effectively partition the vast number of microbial cells from the underlying epithelium. The normal functionality and biochemistry of this mucus barrier appears to be lost in diseases of the colorectal mucosa. Germ-free animal studies have highlighted the necessity of the presence of the colonic microflora to drive the maturation of the colonic mucosa and normal mucus production. A number of by-products of the microflora have been suggested to be key luminal drivers of colonic mucus secretion.

  17. In vivo effects on the intestinal microflora of Physalis alkekengi var. francheti extracts.

    Science.gov (United States)

    Li, Xinli; Zhang, Cuili; Li, Weiling; Wu, Dachang; Liu, Jianjun; Tang, Li; Xin, Yi

    2013-06-01

    This study aimed to investigate the effects on the intestinal microflora balance of Physalis alkekengi var. francheti extracts (PE) using in vivo mouse model. Luteolin-7-O-β-glycoside, Physalin J, Physalin D, and Physalin P were isolated from PE extracts and identified. Bacteroides, Lactobacillus, Helicobacter, Prevotella, Odoribacter and Oribacterium were detected as dominant organisms in the intestinal tract of mice by denaturing gradient gel electrophoresis (DGGE) analysis. The quality and quantity of Lactobacillus genus increased significantly with increasing concentration of PE. This study shows that the intestinal microflora balance could be improved by PE, and thus, it has the significant potential to be used as a natural agent for restoring the intestinal microflora balance.

  18. Synergistic effect of ionizing radiation on chemical disinfectant treatments for reduction of natural microflora on seafood

    Science.gov (United States)

    Kim, Hyunjoo; Ha, Ji-Hyoung; Lee, Ju-Woon; Jo, Cheorun; Ha, Sang-Do

    2012-08-01

    The purpose of this study was to determine whether combined treatments would produce synergistic disinfection effects on seafood products such as mussel and squid compared with single treatments. We investigated the bactericidal effects of chlorine and ionizing radiation on the natural microflora of mussel and squid. Total aerobic bacteria initially ranged from 102 to 104 Log CFU/g. More than 100 ppm of chlorine and irradiation at 1 kGy were sufficient to reduce the total aerobic bacteria on mussel and squid to a level lower than detection limit (10 CFU/g). Synergistic effects against natural microflora were observed for all combined treatment. These results suggest that a significant synergistic benefit results from combine chlorine-ionizing radiation treatment against natural microflora on mussel and squid.

  19. Dissimilar Regulation of Antimicrobial Proteins in the Midgut of Spodoptera exigua Larvae Challenged with Bacillus thuringiensis Toxins or Baculovirus

    Science.gov (United States)

    Crava, Cristina M.; Jakubowska, Agata K.; Escriche, Baltasar; Herrero, Salvador; Bel, Yolanda

    2015-01-01

    Antimicrobial peptides (AMPs) and lysozymes are the main effectors of the insect immune system, and they are involved in both local and systemic responses. Among local responses, midgut immune reaction plays an important role in fighting pathogens that reach the insect body through the oral route, as do many microorganisms used in pest control. Under this point of view, understanding how insects defend themselves locally during the first phases of infections caused by food-borne pathogens is important to further improve microbial control strategies. In the present study, we analyzed the transcriptional response of AMPs and lysozymes in the midgut of Spodoptera exigua (Lepidoptera: Noctuidae), a polyphagous pest that is commonly controlled by products based on Bacillus thuringiensis (Bt) or baculovirus. First, we comprehensively characterized the transcripts encoding AMPs and lysozymes expressed in S. exigua larval midgut, identifying 35 transcripts that represent the S. exigua arsenal against microbial infection. Secondly, we analyzed their expression in the midgut after ingestion of sub-lethal doses of two different pore-forming B. thuringiensis toxins, Cry1Ca and Vip3Aa, and the S. exigua nucleopolyhedrovirus (SeMNPV). We observed that both Bt toxins triggered a similar, wide and in some cases high transcriptional activation of genes encoding AMPs and lysozymes, which was not reflected in the activation of the classical systemic immune-marker phenoloxidase in hemolymph. Baculovirus ingestion resulted in the opposed reaction: Almost all transcripts coding for AMPs and lysozymes were down-regulated or not induced 96 hours post infection. Our results shed light on midgut response to different virulence factors or pathogens used nowadays as microbial control agents and point out the importance of the midgut immune response contribution to the larval immunity. PMID:25993013

  20. Dissimilar Regulation of Antimicrobial Proteins in the Midgut of Spodoptera exigua Larvae Challenged with Bacillus thuringiensis Toxins or Baculovirus.

    Science.gov (United States)

    Crava, Cristina M; Jakubowska, Agata K; Escriche, Baltasar; Herrero, Salvador; Bel, Yolanda

    2015-01-01

    Antimicrobial peptides (AMPs) and lysozymes are the main effectors of the insect immune system, and they are involved in both local and systemic responses. Among local responses, midgut immune reaction plays an important role in fighting pathogens that reach the insect body through the oral route, as do many microorganisms used in pest control. Under this point of view, understanding how insects defend themselves locally during the first phases of infections caused by food-borne pathogens is important to further improve microbial control strategies. In the present study, we analyzed the transcriptional response of AMPs and lysozymes in the midgut of Spodoptera exigua (Lepidoptera: Noctuidae), a polyphagous pest that is commonly controlled by products based on Bacillus thuringiensis (Bt) or baculovirus. First, we comprehensively characterized the transcripts encoding AMPs and lysozymes expressed in S. exigua larval midgut, identifying 35 transcripts that represent the S. exigua arsenal against microbial infection. Secondly, we analyzed their expression in the midgut after ingestion of sub-lethal doses of two different pore-forming B. thuringiensis toxins, Cry1Ca and Vip3Aa, and the S. exigua nucleopolyhedrovirus (SeMNPV). We observed that both Bt toxins triggered a similar, wide and in some cases high transcriptional activation of genes encoding AMPs and lysozymes, which was not reflected in the activation of the classical systemic immune-marker phenoloxidase in hemolymph. Baculovirus ingestion resulted in the opposed reaction: Almost all transcripts coding for AMPs and lysozymes were down-regulated or not induced 96 hours post infection. Our results shed light on midgut response to different virulence factors or pathogens used nowadays as microbial control agents and point out the importance of the midgut immune response contribution to the larval immunity.

  1. DNA sequencing reveals the midgut microbiota of diamondback moth, Plutella xylostella (L. and a possible relationship with insecticide resistance.

    Directory of Open Access Journals (Sweden)

    Xiaofeng Xia

    Full Text Available BACKGROUND: Insect midgut microbiota is important in host nutrition, development and immune response. Recent studies indicate possible links between insect gut microbiota and resistance to biological and chemical toxins. Studies of this phenomenon and symbionts in general have been hampered by difficulties in culture-based approach. In the present study, DNA sequencing was used to examine the midgut microbiota of diamondback moth (DBM, Plutella xylostella (L., a destructive pest that attacks cruciferous crops worldwide. Its ability to develop resistance to many types of synthetic insecticide and even Bacillus thuringiensis toxins makes it an important species to study. METHODOLOGY/PRINCIPAL FINDINGS: Bacteria of the DBM larval midgut in a susceptible and two insecticide (chlorpyrifos and fipronil resistant lines were examined by Illumina sequencing sampled from an insect generation that was not exposed to insecticide. This revealed that more than 97% of the bacteria were from three orders: Enterobacteriales, Vibrionales and Lactobacillales. Both insecticide-resistant lines had more Lactobacillales and the much scarcer taxa Pseudomonadales and Xanthomonadales with fewer Enterobacteriales compared with the susceptible strain. Consistent with this, a second study observed an increase in the proportion of Lactobacillales in the midgut of DBM individuals from a generation treated with insecticides. CONCLUSIONS/SIGNIFICANCE: This is the first report of high-throughput DNA sequencing of the entire microbiota of DBM. It reveals differences related to inter- and intra-generational exposure to insecticides. Differences in the midgut microbiota among susceptible and insecticide-resistant lines are independent of insecticide exposure in the sampled generations. While this is consistent with the hypothesis that Lactobacillales or other scarcer taxa play a role in conferring DBM insecticide resistance, further studies are necessary to rule out other

  2. Dissimilar Regulation of Antimicrobial Proteins in the Midgut of Spodoptera exigua Larvae Challenged with Bacillus thuringiensis Toxins or Baculovirus.

    Directory of Open Access Journals (Sweden)

    Cristina M Crava

    Full Text Available Antimicrobial peptides (AMPs and lysozymes are the main effectors of the insect immune system, and they are involved in both local and systemic responses. Among local responses, midgut immune reaction plays an important role in fighting pathogens that reach the insect body through the oral route, as do many microorganisms used in pest control. Under this point of view, understanding how insects defend themselves locally during the first phases of infections caused by food-borne pathogens is important to further improve microbial control strategies. In the present study, we analyzed the transcriptional response of AMPs and lysozymes in the midgut of Spodoptera exigua (Lepidoptera: Noctuidae, a polyphagous pest that is commonly controlled by products based on Bacillus thuringiensis (Bt or baculovirus. First, we comprehensively characterized the transcripts encoding AMPs and lysozymes expressed in S. exigua larval midgut, identifying 35 transcripts that represent the S. exigua arsenal against microbial infection. Secondly, we analyzed their expression in the midgut after ingestion of sub-lethal doses of two different pore-forming B. thuringiensis toxins, Cry1Ca and Vip3Aa, and the S. exigua nucleopolyhedrovirus (SeMNPV. We observed that both Bt toxins triggered a similar, wide and in some cases high transcriptional activation of genes encoding AMPs and lysozymes, which was not reflected in the activation of the classical systemic immune-marker phenoloxidase in hemolymph. Baculovirus ingestion resulted in the opposed reaction: Almost all transcripts coding for AMPs and lysozymes were down-regulated or not induced 96 hours post infection. Our results shed light on midgut response to different virulence factors or pathogens used nowadays as microbial control agents and point out the importance of the midgut immune response contribution to the larval immunity.

  3. Gene expression analysis of the endosymbiont-bearing midgut tissue during ontogeny of the carpenter ant Camponotus floridanus.

    Science.gov (United States)

    Ratzka, Carolin; Gross, Roy; Feldhaar, Heike

    2013-06-01

    Insects have frequently evolved mutualistic relationships with extracellular and/or intracellular bacterial endosymbionts. Infection with endosymbionts seems to affect several cellular functions of the host such as immune pathways, oxidative stress regulation and autophagy. Our current knowledge about specific host factors leading to endosymbiont tolerance and/or control is still scarce and is based on very few associations between insect hosts and bacteria only. Camponotus floridanus ants harbour the obligate intracellular bacterium Blochmannia floridanus within specialized midgut cells called bacteriocytes. The number of Blochmannia endosymbionts within the midgut tissue increases strongly during host development and reaches a maximum at the late pupal stage, where the entire midgut is transformed into a symbiotic organ. After eclosion of workers the number of Blochmannia strongly decreases again. We chose 15 candidate genes from C. floridanus likely to be involved in host-symbiont interactions based on their significant homology to previously investigated symbiosis-relevant genes from other insects. We determined the expression of these genes in the endosymbiont-bearing midgut tissue in comparison to the residual body tissue at different developmental stages of C. floridanus in order to reveal changes in gene expression correlating with changes in endosymbiont number per host. Strikingly, two pattern recognition receptors (amidase PGRP-LB and PGRP-SC2) were highly expressed in the midgut tissue at the pupal stage, potentially down-modulating the IMD pathway to enable endosymbiont tolerance. Moreover, we investigated the immune gene expression in response to bacterial challenge at the pupal stage. Results showed that the midgut tissue differs in expression pattern in contrast to the residual body. Our results support a key role for amidase PGRPs, especially PGRP-LB, in regulation of the immune response towards endosymbionts in C. floridanus and suggest an

  4. A GPI-anchored alkaline phosphatase is a functional midgut receptor of Cry11Aa toxin in Aedes aegypti larvae.

    Science.gov (United States)

    Fernandez, Luisa E; Aimanova, Karlygash G; Gill, Sarjeet S; Bravo, Alejandra; Soberón, Mario

    2006-02-15

    A 65 kDa GPI (glycosylphosphatidyl-inositol)-anchored ALP (alkaline phosphatase) was characterized as a functional receptor of the Bacillus thuringiensis subsp. israelensis Cry11Aa toxin in Aedes aegypti midgut cells. Two (a 100 kDa and a 65 kDa) GPI-anchored proteins that bound Cry11Aa toxin were preferentially extracted after treatment of BBMV (brush boder membrane vesicles) from Ae. aegypti midgut epithelia with phospholipase C. The 65 kDa protein was further purified by toxin affinity chromatography. The 65 kDa protein showed ALP activity. The peptide-displaying phages (P1.BBMV and P8.BBMV) that bound to the 65 kDa GPI-ALP (GPI-anchored ALP) and competed with the Cry11Aa toxin to bind to BBMV were isolated by selecting BBMV-binding peptide-phages by biopanning. GPI-ALP was shown to be preferentially distributed in Ae. aegypti in the posterior part of the midgut and in the caeca, by using P1.BBMV binding to fixed midgut tissue sections to determine the location of GPI-ALP. Cry11Aa binds to the same regions of the midgut and competed with P1.BBMV and P8.BBMV to bind to BBMV. The importance of this interaction was demonstrated by the in vivo attenuation of Cry11Aa toxicity in the presence of these phages. Our results shows that GPI-ALP is an important receptor molecule involved in Cry11Aa interaction with midgut cells and toxicity to Ae. aegypti larvae.

  5. Microflora of conjunctiva in children and its sensitivity and resistance to antibacterial drugs

    Directory of Open Access Journals (Sweden)

    T. N. Vorontsova

    2012-01-01

    Full Text Available Purpose: Investigation of microflora of conjunctiva and its resistance to antibacterial drugs in healthy children and patients with various inflammatory eye diseases.Methods: We examined 402 children (421 eyes in the age from 1 month till 17 years: 62 healthy children (70 eyes and 340 pa- tients with different inflammatory diseases of anterior segment of eye (351 eyes. the smear was done in all children for plating and definition of sensitivity of microflora to antibacterial drugs by method of diffusion to agar.Results: the plating was positive even in 72.9% of healthy children who entered the hospital for the planned surgery. Most often we revealed Staphylococcus epidermidis (44.3%, Staphylococcus aureus (12.8%, Streptococcus faecalis (5.7% and Enterobacter (2.9%. In children with inflammatory diseases Staphylococcus epidermidis and Staphylococcus aureus (62.6% were found fre- quently. the analysis of data showed high level of resistance of all microflora to aminoglycosides (neomycin 37.8% and tobramycin 32.7% and chloramphenicol — 37.1%. the lowest resistance of all microflora was registered to levofloxacin (11.1% and ciprofloxacin (10.5%. In gram-negative microflora we revealed the maximal sensitivity to ciprofloxacin, in gram-positive — to levofloxacin.We detected the maximal resistance of microflora to ampicillin (66.1%, and minimal — to cephalosporines (4.5% among the antibiotics of systemic application.Conclusion: the findings allow us to recommend drops containing levofloxacin (Signicef for clinical practice in pediatric ophthalmology. 

  6. Maternal vaginal microflora during pregnancy and the risk of asthma hospitalization and use of antiasthma medication in early childhood

    DEFF Research Database (Denmark)

    Benn, Christine Stabell; Thorsen, Poul; Jensen, Jørgen Skov;

    2002-01-01

    BACKGROUND: Infants with wheezing and allergic diseases have a microflora that differs from that of healthy infants. The fetus acquires microorganisms during birth when exposed to the maternal vaginal microflora. It is therefore conceivable that the maternal vaginal microflora might influence...... the establishment of the infant flora and, as a consequence, the development of wheezing and allergic diseases. OBJECTIVE: We sought to study the associations between the composition of the maternal vaginal microflora and the development of wheezing and asthma in childhood. METHODS: We performed a population....... Maternal colonization with staphylococci (OR, 2.2; 95% CI, 1.4-3.4) and use of antibiotics in pregnancy (OR, 1.7; 95% CI, 1.1-2.6) were associated with asthma during the fifth year of life. CONCLUSION: The composition of the maternal vaginal micro-flora might be associated with wheezing and asthma...

  7. ROLE OF THE MICROFLORA IN DISTAL INTESTINAL TRACT BY MAINTAINING OXALATE HOMEOSTASIS

    Directory of Open Access Journals (Sweden)

    Osolodchenko T.P.

    2015-05-01

    Full Text Available Human intestinal microflora is part of the human body and performs numerous function. Considerable research interest is in the field of probiotics for the prevention of kidney stones, which is one of the most common urological diseases.Urolithiasis is one of the most common urological diseases. This is polyetiological disease congenital and acquired character with complex physical and chemical processes that occur not only in the urinary system, but also the whole body. None of the treatments does not guarantee full recovery of the patient and often leads to relapse. The open methods of removal stones yield news minimally invasive the technologys. Development of stone formation depends on the presence of many factors, metabolic disorders, chronic urinary tract infections, genetic disorders and more. Most have the following metabolic disorders as hypercalciuria, hiperurikuria, hipotsytraturia , hyperoxaluria and hipomahniuria. Among all types of urolithiasis kaltsiyoksalatnyy ranked first in the prevalence rate - about 75.0 - 85.0 % of cases. Dietary restriction by oxalates іs the unreliable method of preventing disease. Although there is evidence for the growth inhibition normobiocenosis representatives, which in turn enhances the absorption of salts of oxalic acid oxalate in the application of sodium , magnesium and cobalt in their intragastric administration. Recently published many papers on the impact on the level of oxalate intestinal microflora. The first publications appeared on the influence of gram-negative obligate anaerobes O. formigenes the concentration of oxalate in the urine. This anaerobic bacteria living in the colon, its prevalence - 46.0 % - 77.0 % of the adult population. O. formigenes reveals the symbiotic interaction with the human body by reducing absorption of oxalate in the intestinal cavity with subsequent decrease in their concentration in plasma and urine. O. formigenes has two key enzymes - oksalyl

  8. Morphological changes in the midgut of Aedes aegypti L. (Diptera: Culicidae) larvae following exposure to an Annona coriacea (Magnoliales: Annonaceae) extract.

    Science.gov (United States)

    Costa, M S; Pinheiro, D O; Serrão, J E; Pereira, M J B

    2012-08-01

    Bioinsecticides are important in the control of disease vectors, but data regarding their physiological effects on target insects are incomplete. This study describes morphological changes that occur in the midgut of third instar Aedes aegypti L. (Diptera: Culicidae) following treatment with a methanolic extract of Annona coriacea (Magnoliales: Annonaceae). Dissected midguts were subdivided into anterior and posterior regions and analyzed by light and scanning electron microscopy. Insects exposed to the extract displayed intense, destructive cytoplasmic vacuolization in columnar and regenerative midgut cells. The apical surfaces of columnar cells exhibited cytoplasmic protrusions oriented toward the lumen, suggesting that these cells could be involved in apocrine secretory processes and/or apoptosis. We report that A. coriacea extracts induced morphological alterations in the midgut of A. aegypti midgut larvae, supporting the use of plant extracts for control of the dengue vector.

  9. [Species composition of anaerobic microflora in parodontal pocket depending upon disease stage].

    Science.gov (United States)

    Zyrianova, N V; Grigor'ian, A S; Grudianov, A I; Frolova, O A; Shil'nikova, I I; Kobozev, M I

    2009-01-01

    With the help of polymerase chain reaction (PCR) the dynamic of species composition of anaerobic microflora in cases of generalized parodontitis was established. It was detected that disease severity increase was followed by the increase of the number of anaerobic microflora species in parodontal pocket; at that it was impossible to connect the presence of some determined type of microorganism with the inflammatory parodontal process intensity. It was shown that proteins fimbrilin and gingipain were not the only parodontitis pathogenic factors although the first one (fimbrilin) could be connected with aggressive disease flow. The suggested PCR scheme could be useful for early disease stage diagnostic and substantiation of antimicrobial therapy method selection.

  10. The effect of lacto- and bifidobacteria compositions on the vaginal microflora in cases of intravaginal staphylococcosis.

    Science.gov (United States)

    Babenko, L P; Lazarenko, L M; Shynkarenko, L M; Mokrozub, V V; Pidgorskyi, V S; Spivak, M Ja

    2012-01-01

    The effect of intravaginal injection of Lactobacillus casei IMV B-7280, Lactobacillus acidophilus IMV B-7279, Bifidobacterium animalis VKL and Bifidobacterium animalis VKB strains in various compositions on the range of microflora of the urogenital tract on the model of experimental intravaginal staphylococcosis of mice was determined. It was established that under the influence of various compositions of these strains changes in microflora spectrum occurred: the number of representatives of normoflora increased and the number of pathogenic microorganisms, including staphylococci, streptococci, coliform bacteria and fungi, significantly decreased. It was determined that strains of lacto- and bifidobacteria, that were studied, are prospective components of future probiotic drugs efficient in treating staphylococcosis.

  11. Impact on bacterial community in midguts of the Asian corn borer larvae by transgenic Trichoderma strain overexpressing a heterologous chit42 gene with chitin-binding domain.

    Directory of Open Access Journals (Sweden)

    Yingying Li

    Full Text Available This paper is the first report of the impact on the bacterial community in the midgut of the Asian corn borer (Ostrinia furnacalis by the chitinase from the transgenic Trichoderma strain. In this study, we detected a change of the bacterial community in the midgut of the fourth instar larvae by using a culture-independent method. Results suggested that Proteobacteria and Firmicutes were the most highly represented phyla, being present in all the midgut bacterial communities. The observed species richness was simple, ranging from four to five of all the 16S rRNA clone libraries. When using Trichoderma fermentation liquids as additives, the percentages of the dominant flora in the total bacterial community in larval midgut changed significantly. The community of the genus Ochrobactrum in the midgut decreased significantly when the larvae were fed with the fermentation liquids of the transgenic Trichoderma strain Mc4. However, the Enterococcus community increased and then occupied the vacated niche of the Ochrobactrum members. Furthermore, the Shannon-Wiener (H and the Simpson (1-D indexes of the larval midgut bacterial library treated by feeding fermentation liquids of the transgenic Trichoderma strain Mc4 was the lowest compared with the culture medium, fermentation liquids of the wild type strain T30, and the sterile artificial diet. The Enterococcus sp. strain was isolated and characterized from the healthy larvae midgut of the Asian corn borer. An infection study of the Asian corn borer larvae using Enterococcus sp. ACB-1 revealed that a correlation existed between the increased Enterococcus community in the larval midgut and larval mortality. These results demonstrated that the transgenic Trichoderma strain could affect the composition of the midgut bacterial community. The change of the midgut bacterial community might be viewed as one of the factors resulting in the increased mortality of the Asian corn borer larvae.

  12. Impact on bacterial community in midguts of the Asian corn borer larvae by transgenic Trichoderma strain overexpressing a heterologous chit42 gene with chitin-binding domain.

    Science.gov (United States)

    Li, Yingying; Fu, Kehe; Gao, Shigang; Wu, Qiong; Fan, Lili; Li, Yaqian; Chen, Jie

    2013-01-01

    This paper is the first report of the impact on the bacterial community in the midgut of the Asian corn borer (Ostrinia furnacalis) by the chitinase from the transgenic Trichoderma strain. In this study, we detected a change of the bacterial community in the midgut of the fourth instar larvae by using a culture-independent method. Results suggested that Proteobacteria and Firmicutes were the most highly represented phyla, being present in all the midgut bacterial communities. The observed species richness was simple, ranging from four to five of all the 16S rRNA clone libraries. When using Trichoderma fermentation liquids as additives, the percentages of the dominant flora in the total bacterial community in larval midgut changed significantly. The community of the genus Ochrobactrum in the midgut decreased significantly when the larvae were fed with the fermentation liquids of the transgenic Trichoderma strain Mc4. However, the Enterococcus community increased and then occupied the vacated niche of the Ochrobactrum members. Furthermore, the Shannon-Wiener (H) and the Simpson (1-D) indexes of the larval midgut bacterial library treated by feeding fermentation liquids of the transgenic Trichoderma strain Mc4 was the lowest compared with the culture medium, fermentation liquids of the wild type strain T30, and the sterile artificial diet. The Enterococcus sp. strain was isolated and characterized from the healthy larvae midgut of the Asian corn borer. An infection study of the Asian corn borer larvae using Enterococcus sp. ACB-1 revealed that a correlation existed between the increased Enterococcus community in the larval midgut and larval mortality. These results demonstrated that the transgenic Trichoderma strain could affect the composition of the midgut bacterial community. The change of the midgut bacterial community might be viewed as one of the factors resulting in the increased mortality of the Asian corn borer larvae.

  13. Contribution of midgut bacteria to blood digestion and egg production in aedes aegypti (diptera: culicidae (L.

    Directory of Open Access Journals (Sweden)

    Pimenta Paulo FP

    2011-06-01

    Full Text Available Abstract Background The insect gut harbors a variety of microorganisms that probably exceed the number of cells in insects themselves. These microorganisms can live and multiply in the insect, contributing to digestion, nutrition, and development of their host. Recent studies have shown that midgut bacteria appear to strengthen the mosquito's immune system and indirectly enhance protection from invading pathogens. Nevertheless, the physiological significance of these bacteria for mosquitoes has not been established to date. In this study, oral administration of antibiotics was employed in order to examine the contribution of gut bacteria to blood digestion and fecundity in Aedes aegypti. Results The antibiotics carbenicillin, tetracycline, spectinomycin, gentamycin and kanamycin, were individually offered to female mosquitoes. Treatment of female mosquitoes with antibiotics affected the lysis of red blood cells (RBCs, retarded the digestion of blood proteins and reduced egg production. In addition, antibiotics did not affect the survival of mosquitoes. Mosquito fertility was restored in the second gonotrophic cycle after suspension of the antibiotic treatment, showing that the negative effects of antibiotics in blood digestion and egg production in the first gonotrophic cycle were reversible. Conclusions The reduction of bacteria affected RBC lysis, subsequently retarded protein digestion, deprived mosquito from essential nutrients and, finally, oocyte maturation was affected, resulting in the production of fewer viable eggs. These results indicate that Ae. aegypti and its midgut bacteria work in synergism to digest a blood meal. Our findings open new possibilities to investigate Ae. aegypti-associated bacteria as targets for mosquito control strategies.

  14. Posterior midgut epithelial cells differ in their organization of the membrane skeleton from other drosophila epithelia.

    Science.gov (United States)

    Baumann, O

    2001-11-01

    In epithelial cells, the various components of the membrane skeleton are segregated within specialized subregions of the plasma membrane, thus contributing to the development and stabilization of cell surface polarity. It has previously been shown that, in various Drosophila epithelia, the membrane skeleton components ankyrin and alphabeta-spectrin reside at the lateral surface, whereas alphabeta(H)-spectrin is restricted to the apical domain. By use of confocal immunofluorescence microscopy, the present study characterizes the membrane skeleton of epithelial cells in the posterior midgut, leading to a number of unexpected results. First, ankyrin and alphabeta-spectrin are not detected on the entire lateral surface but appear to be restricted to the apicolateral area, codistributing with fasciclin III at smooth septate junctions. The presumptive ankyrin-binding proteins neuroglian and Na(+),K(+)-ATPase, however, do not colocalize with ankyrin. Second, alphabeta(H)-spectrin is enriched at the apical domain but is also present in lower amounts on the entire lateral surface, colocalizing apicolaterally with ankyrin/alphabeta-spectrin. Finally, despite the absence of zonulae adherentes, F-actin, beta(H)-spectrin, and nonmuscle myosin-II are enriched in the midlateral region. Thus, the model established for the organization of the membrane skeleton in Drosophila epithelia does not hold for the posterior midgut, and there is quite some variability between the different epithelia with respect to the organization of the membrane skeleton.

  15. Exosome Secretion by the Parasitic Protozoan Leishmania within the Sand Fly Midgut

    Directory of Open Access Journals (Sweden)

    Vanessa Diniz Atayde

    2015-11-01

    Full Text Available Despite several studies describing the secretion of exosomes by Leishmania in vitro, observation of their formation and release in vivo has remained a major challenge. Herein, we show that Leishmania constitutively secretes exosomes within the lumen of the sand fly midgut through a mechanism homologous to the mammalian pathway. Through egestion experiments, we demonstrate that Leishmania exosomes are part of the sand fly inoculum and are co-egested with the parasite during the insect’s bite, possibly influencing the host infectious process. Indeed, co-inoculation of mice footpads with L. major plus midgut-isolated or in-vitro-isolated L. major exosomes resulted in a significant increase in footpad swelling. Notably, co-injections produced exacerbated lesions through overinduction of inflammatory cytokines, in particular IL-17a. Our data indicate that Leishmania exosomes are an integral part of the parasite’s infectious life cycle, and we propose to add these vesicles to the repertoire of virulence factors associated with vector-transmitted infections.

  16. The Bacillus thuringiensis vegetative insecticidal protein Vip3A lyses midgut epithelium cells of susceptible insects.

    Science.gov (United States)

    Yu, C G; Mullins, M A; Warren, G W; Koziel, M G; Estruch, J J

    1997-02-01

    The Vip3A protein is a member of a newly discovered class of vegetative insecticidal proteins with activity against a broad spectrum of lepidopteran insects. Histopathological observations indicate that Vip3A ingestion by susceptible insects such as the black cutworm (Agrotis ipsilon) and fall armyworm (Spodoptera frugiperda) causes gut paralysis at concentrations as low as 4 ng/cm2 of diet and complete lysis of gut epithelium cells resulting in larval death at concentrations above 40 ng/cm2. The European corn borer (Ostrinia nubilalis), a nonsusceptible insect, does not develop any pathology upon ingesting Vip3A. While proteolytic processing of the Vip3A protein by midgut fluids obtained from susceptible and nonsusceptible insects is comparable, in vivo immunolocalization studies show that Vip3a binding is restricted to gut cells of susceptible insects. Therefore, the insect host range for Vip3A seems to be determined by its ability to bind gut cells. These results indicate that midgut epithelium cells of susceptible insects are the primary target for the Vip3A insecticidal protein and that their subsequent lysis is the primary mechanism of lethality. Disruption of gut cells appears to be the strategy adopted by the most effective insecticidal proteins.

  17. "Comparison of Midgut Hemagglutination Activity in Three Different Geographical Populations of Anopheles stephensi"

    Directory of Open Access Journals (Sweden)

    HR Basseri

    2004-08-01

    Full Text Available Lectins that agglutinate red blood cells (RBCs were demonstrated in Anopheles stephensi mosquito midgut extracts using human (four groups: A, B, AB and O, RH+ rat, sheep and rabbit blood cells. Only rabbit RBCs showed agglutination reaction against the midgut extracts. Significant differences in hemagglutinin titers and carbohydrate specifity were detected between male and female mosquitoes as well as among three different geographical populations of Anopheles stephensi from south of Iran. Overall agglutinin levels were increased following a blood meal. The highest hemagglintination titers were due to Kazerun population. All hemagglutination assays were versus rabbit RBCS. A significant difference was detected among the number of egg-float ridges. Iranshahr population was different from Bandar-abbas and Kazerun population in egg-float ridges number. Bandr-abbas population was in the intermediate category. Iranshahr population fell between mysoransis and intermediate group and Kazerun population was between intermediate and type form. This study presents the first report on the occurrence of heterogeneous anti Rabbit RBC agglutinins in the midget extracts of the different geographical populations of An.stephensi with the sugar – binding specificities. The sugar- inhibition pattern was different between & within geographical population of An.stephensi.

  18. Malaria parasites form filamentous cell-to-cell connections during reproduction in the mosquito midgut

    Institute of Scientific and Technical Information of China (English)

    Ingrid Rupp; Gabriele Pradel; Ludmilla Sologub; Kim C Williamson; Matthias Scheuermayer; Luc Reininger; Christian Doerig; Saliha Eksi; Davy U Kombilaa; Matthias Frank

    2011-01-01

    Physical contact is important for the interaction between animal cells, but it can represent a major challenge for protists like malaria parasites. Recently, novel filamentous cell-cell contacts have been identified in different types of eukaryotic cells and termed nanotubes due to their morphological appearance. Nanotubes represent small dynamic membranous extensions that consist of F-actin and are considered an ancient feature evolved by eukaryotic cells to establish contact for communication. We here describe similar tubular structures in the malaria pathogen Plasmodium falciparum, which emerge from the surfaces of the forming gametes upon gametocyte activation in the mosquito midgut. The filaments can exhibit a length of>100 μm and contain the F-actin isoform actin 2. They actively form within a few minutes after gametocyte activation and persist until the zygote transforms into the ookinete. The filaments originate from the parasite plasma membrane, are close ended and express adhesion proteins on their surfaces that are typically found in gametes, like Pfs230, Pfs48/45 or Pfs25, but not the zygote surface protein Pfs28. We show that these tubular structures represent long-distance cell-to-cell connections between sexual stage parasites and demonstrate that they meet the characteristics of nanotubes. We propose that malaria parasites utilize these adhesive "nanotubes" in order to facilitate intercellular contact between gametes during reproduction in the mosquito midgut.

  19. ESTABLISHMENT OF PRIMARY AND CONTINUOUS CULTURE OF MIDGUT EPITHELIAL CELLS FROM AEDES AEGYPTI(DIPTERA:CULICIDAE) IN VITRO%埃及伊蚊中肠上皮细胞的原代培养和细胞的持续培养

    Institute of Scientific and Technical Information of China (English)

    公茂庆; 黄勇; 谭文彬; Brian D. For; John C. Beier

    2003-01-01

    目的探讨蚊虫中肠上皮细胞的培养,建立细胞的原代培养和细胞持续培养. 方法用含抗生素的10%蔗糖水喂养新羽化出的埃及伊蚊成蚊,3 d后,在无菌的条件下解剖中肠,用含抗生素的平衡盐溶液洗3次,放入Grace's培养基中(含有7%小牛血清,0.5%庆大霉素,1 mg/ml抗酵母菌素和6 μl/ml 复合维生素),25℃培养箱中培养. 结果培养3 d后,95%的细胞生长良好,7 d后,大约25%的细胞死亡.第3 d的细胞死亡率明显低于以后的任何时间(P0.05).成功地建立起中肠上皮细胞的原代培养并能持续存活40多天,保持细胞的连续培养. 结论所发展的埃及伊蚊中肠上皮细胞的原代培养和细胞的持续培养的方法是成功的.%Objective To investigate the growth and development of the adult mosquito midgut, to develop primary midgut culture. Methods After emerging adult mosquitoes were fed for 3 consecutive days on antibiotic solution, midguts dissected under sterile condition were kept in Grace's medium containing 7% fetal bovine serum, 0.5% gentamycin, 1 mg/ml yeastolate and 6 μl/ml vitamin premix to incubate at 25℃. Results The cells recovered from the midgut are healthy with about 95% viability on day 3. Approximately 25% of the cells died one week later, but those that survive stay in a stable condition during 35 days. There are significantly fewer dead cells on day 3 (P0.05). Primary cultures that would survive up to over a 40-day incubation period were successfully developed and continuous cultures were established. Conclusion The procedure that would facilitate the culture of midgut epithelial cells from Aedes aegypti adult mosquitoes was developed.

  20. Associations of GI microflora with campylobacter status in commercial broiler chickens

    Science.gov (United States)

    Introduction: Nextgeneration DNA sequencing is a powerful tool for food safety. One potential use for this technology is to compare the microflora of food animals hosting foodborne pathogens to pathogen-free animals to better understand the ecology of foodborne pathogens to design possible intervent...

  1. Benzene bioremediation using cow dung microflora in two phase partitioning bioreactor

    Energy Technology Data Exchange (ETDEWEB)

    Singh, Dipty [Environmental Biotechnology Laboratory, Department of Life Sciences, University of Mumbai, Vidyanagari Campus, Santacruz (E), Mumbai-400 098 (India); Fulekar, M.H., E-mail: mhfulekar@yahoo.com [Environmental Biotechnology Laboratory, Department of Life Sciences, University of Mumbai, Vidyanagari Campus, Santacruz (E), Mumbai-400 098 (India)

    2010-03-15

    Bioremediation of benzene has been carried out using cow dung microflora in a bioreactor. The bioremediation of benzene under the influence of cow dung microflora was found to be 100% and 67.5%, at initial concentrations of 100 mg/l and 250 mg/l within 72 h and 168 h respectively; where as at higher concentration (500 mg/l), benzene was found to be inhibitory. Hence the two phase partitioning bioreactor (TPPB) has been designed and developed to carryout biodegradation at higher concentration. In TPPB 5000 mg/l benzene was biodegraded up to 50.17% over a period of 168 h. Further the Pseudomonas putida MHF 7109 was isolated from cow dung microflora as potential benzene degrader and its ability to degrade benzene at various concentrations was evaluated. The data indicates 100%, 81% and 65% degradation at the concentrations of 50 mg/l, 100 mg/l, 250 mg/l within the time period of 24 h, 96 h and 168 h respectively. The GC-MS data also shows the presence of catechol and 2-hydroxymuconic semialdehyde, which confirms the established pathway of benzene biodegradation. The present research proves the potential of cow dung microflora as a source of biomass for benzene biodegradation in TPPB.

  2. Effects of a Campylobacter jejuni infection on the development of the intestinal microflora of broiler chickens

    DEFF Research Database (Denmark)

    Johansen, C. H.; Friis-Holm, Lotte Bjerrum; Finster, K.

    2006-01-01

    The effect of a Campylobacter jejuni colonization on the development of the microflora of the cecum and the ileum of broiler chickens was studied using molecular methods. The infection did affect the development and complexity of the microbial Communities of the ceca, but we found no permanent...

  3. Intestinal microflora molecular markers of spleen-deficient rats and evaluation of traditional Chinese drugs

    Institute of Scientific and Technical Information of China (English)

    Ying Peng; Zhuo Wang; Yuan Lu; Chun-Fu Wu; Jing-Yu Yang; Xiao-Bo Li

    2009-01-01

    AIM: To find a rapid and efficient analysis method of gastrointestinal microflora in Pi-deficient (spleendeficient) rats and to evaluate traditional Chinese drugs. METHODS: Enterobacterial repetitive intergenic consensus-PCR (ERIC-PCR) based assay was performed to examine changes of intestinal microflora in two Pi-deficienct animal models and to evaluate the efficacy of four traditional Chinese drugs as well as a probiotic recipe and another therapy in Pi-deficient rats. RESULTS: A molecular marker was identified for Pideficiency in rats. The pharmacodynamic evaluation system, including identified molecular markers (net integral area and abundance of DNA bands), Shannon's index for diversity of intestinal microflora, and Sorenson's pairwise similarity coefficient, was established. The four major clinical recipes of traditional Chinese drugs for Pi-deficiency in rats, especially at their medium dose (equivalence to the clinical dose), produced more pronounced recovery activities in Pi-deficient rats, while higher doses of these recipes did not show a better therapeutic effect but some toxic effects such as perturbation deterioration of intestinal microflora. CONCLUSION: Both fingerprint analysis and identified marker can show Pi-deficiency in rats and its difference after treatment. The identified molecular marker may be applied in screening for the active compounds both in relative traditional Chinese drugs and in pharmacodynamic study of Pi-deficiency in rats.

  4. The effect of higher plant microflora on the microbial landscape of a closed ecosystem

    Science.gov (United States)

    Tirranen, Lyalya; Gitelson, Josef; Borodina, Elena

    2012-07-01

    Having summarized certain data obtained earlier, we defined the aim of this work as an estimation of the effect of higher plant microflora on the microbial landscape of a closed ecosystem (CES). The microflora of such a component as higher plants can influence other system components not only by way of transfer with air and water flows, but also through the direct contact of the crew with the crops cultivated within CES when harvesting, thrashing, using them for food. Involving the higher plant component into the closed system the microorganism diversity and occurrence of microscopic fungi in other components of the closed ecosystem increased. The presence of microscopic fungi, especially on plants and in the air, is potentially dangerous for the health of the system residents. Since the contribution of the higher plant microflora (especially mycoflora) to the microbial landscape of a CES is significant, it is necessary to reduce the microbial flora of the higher plant component and limit its dispersion to other system components. One of the possible measures to limit the higher plant microflora colonization is air purification between components. Reducing the number of microscopic fungi by decreasing the humidity in the system's atmosphere should also be considered.

  5. Early life events influence whole-of-life metabolic health via gut microflora and gut permeability.

    Science.gov (United States)

    Kerr, Caroline A; Grice, Desma M; Tran, Cuong D; Bauer, Denis C; Li, Dongmei; Hendry, Phil; Hannan, Garry N

    2015-01-01

    The capacity of our gut microbial communities to maintain a stable and balanced state, termed 'resilience', in spite of perturbations is vital to our achieving and maintaining optimal health. A loss of microbial resilience is observed in a number of diseases including obesity, diabetes and metabolic syndrome. There are large gaps in our understanding of why an individual's co-evolved microflora consortium fail to develop resilience thereby establishing a trajectory towards poor metabolic health. This review examines the connections between the developing gut microbiota and intestinal barrier function in the neonate, infant and during the first years of life. We propose that the effects of early life events on the gut microflora and permeability, whilst it is in a dynamic and vulnerable state, are fundamental in shaping the microbial consortia's resilience and that it is the maintenance of resilience that is pivotal for metabolic health throughout life. We review the literature supporting this concept suggesting new potential research directions aimed at developing a greater understanding of the longitudinal effects of the gut microflora on metabolic health and potential interventions to recalibrate the 'at risk' infant gut microflora in the direction of enhanced metabolic health.

  6. Suppression of Listeria monocytogenes by the Native Micro-Flora in Teewurst Sausage

    Directory of Open Access Journals (Sweden)

    Michline Brice

    2013-10-01

    Full Text Available Modern consumers are interested in the use of non-chemical methods to control pathogens when heat sterilization is not an option. Such is the case with teewurst sausage, a raw spreadable sausage and a popular German commodity. Although Listeria was not found in teewurst, the optimal microbial growing conditions of teewurst coupled with the ubiquity of L. monocytogenes in nature, makes the possibility of contamination of products very possible. This pilot study was conducted to examine teewurst’s native micro-flora’s ability to suppress the outgrowth of L. monocytogenes at 10 °C using standard plate counts and PCR-DGGE. Traditional plating methods showed L. monocytogenes growth significantly decreased when in competition with the teewurst’s native micro-flora (p < 0.05. The native micro-flora of the teewurst suppressed the overall growth of L. monocytogenes by an average of two logs, under these conditions. Denaturing Gradient Gel Electrophoresis (DGGE amplicons with unique banding patterns were extracted from DGGE gel for identification. Brochothrix thermosphacta and Lactobacillus curvatus were identified as a part of the teewurst’s native micro-flora. Although the native micro-flora did not decrease L. monocytogenes to below limits of detection, it was enough of a decrease to warrant further investigation.

  7. Temperature Fluctuation, Low Salinity, Water Microflora: Risk Factors for WSSV Outbreaks in Penaeus monodon

    NARCIS (Netherlands)

    Tendencia Alapide, E.; Verreth, J.A.J.

    2011-01-01

    White spot syndrome virus (WSSV) has been devastating the shrimp industry for almost a decade. This study compares water parameters, alkalinity, and microflora in three ponds on a farm on Negros Island (Philippines) during two production cycles where WSSV infection resulted in an outbreak in 2006 bu

  8. The four serotypes of dengue recognize the same putative receptors in Aedes aegypti midgut and Ae. albopictus cells

    Directory of Open Access Journals (Sweden)

    Camacho-Nuez Minerva

    2006-10-01

    Full Text Available Abstract Background Dengue viruses (DENV attach to the host cell surface and subsequently enter the cell by receptor-mediated endocytosis. Several primary and low affinity co-receptors for this flavivirus have been identified. However, the presence of these binding molecules on the cell surface does not necessarily render the cell susceptible to infection. Determination of which of them serve as bona fide receptors for this virus in the vector may be relevant to treating DENV infection and in designing control strategies. Results (1 Overlay protein binding assay showed two proteins with molecular masses of 80 and 67 kDa (R80 and R67. (2 Specific antibodies against these two proteins inhibited cell binding and infection. (3 Both proteins were bound by all four serotypes of dengue virus. (4 R80 and R67 were purified by affinity chromatography from Ae. aegypti mosquito midguts and from Ae albopictus C6/36 cells. (5 In addition, a protein with molecular mass of 57 kDa was purified by affinity chromatography from the midgut extracts. (6 R80 and R67 from radiolabeled surface membrane proteins of C6/36 cells were immunoprecipitated by antibodies against Ae. aegypti midgut. Conclusion Our results strongly suggest that R67 and R80 are receptors for the four serotypes of dengue virus in the midgut cells of Ae. aegypti and in C6/36 Ae. albopictus cells.

  9. Different domains of Bacillus thuringiensis delta-endotoxins can bind to insect midgut membrane proteins on ligand blots

    NARCIS (Netherlands)

    Maagd, de R.A.; Klei, van der H.; Bakker, P.L.; Stiekema, W.J.; Bosch, D.

    1996-01-01

    We investigated the role of the constituent domains of the CryIA(b) and CryIA(c) δ-endotoxins in binding to midgut epithelial cell membrane proteins of Spodoptera exigua and Manduca sexta on ligand blots. A collection of wild- type and CryIC-CryIA hybrid toxins was used for this purpose. As demonstr

  10. Effect of silicon on the morphology of the midgut and mandible of tomato leafminer Tuta absoluta (Lepidoptera: Gelechiidae larvae

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    MC dos Santos

    2015-05-01

    Full Text Available Tuta absoluta (Meyrick (Lepidoptera: Gelechiidae is an important insect pest causing serious losses to tomato plantations in Brazil. Some populations of T. absolute are reported to present insecticide resistance resulting in its control failure and the use of alternative control based on silicon, which is clean and sustainable, can reduce pesticide use, increasing fruit quality and protecting the environment. This study evaluated changes in the morphology of the midgut and the mandibles of T. absoluta larvae caused by feeding with compounds containing silicon. Larvae of T. absoluta were fed on tomato leaves with different compounds containing silicon and the histology of the midgut of fourth instar larvae was analyzed. The mandibles of all larval stages were dissected and analysed by scanning electron microscopy. There were no changes in the morphology of the mandibles of the larvae of T. absoluta fed on silicon compared to the control group. Larvae of T. absoluta from the control group and the treatments where the calcium silicate was applied to the soil had not differences in the morphology of the midgut epithelial cells, which had four cell types: digestive, goblet, regenerative and mycetocyte cells. In larvae of T. absoluta obtained from silicon-based treatments applied to the leaves, the midgut epithelium showed detachment of the basal membrane, which can characterize the possible effect of this toxic element to larvae of T. absoluta.

  11. Microbial DNA fingerprinting of human fingerprints: dynamic colonization of fingertip microflora challenges human host inferences for forensic purposes.

    Science.gov (United States)

    Tims, Sebastian; van Wamel, Willem; Endtz, Hubert P; van Belkum, Alex; Kayser, Manfred

    2010-09-01

    Human fingertip microflora is transferred to touched objects and may provide forensically relevant information on individual hosts, such as on geographic origins, if endogenous microbial skin species/strains would be retrievable from physical fingerprints and would carry geographically restricted DNA diversity. We tested the suitability of physical fingerprints for revealing human host information, with geographic inference as example, via microbial DNA fingerprinting. We showed that the transient exogenous fingertip microflora is frequently different from the resident endogenous bacteria of the same individuals. In only 54% of the experiments, the DNA analysis of the transient fingertip microflora allowed the detection of defined, but often not the major, elements of the resident microflora. Although we found microbial persistency in certain individuals, time-wise variation of transient and resident microflora within individuals was also observed when resampling fingerprints after 3 weeks. While microbial species differed considerably in their frequency spectrum between fingerprint samples from volunteers in Europe and southern Asia, there was no clear geographic distinction between Staphylococcus strains in a cluster analysis, although bacterial genotypes did not overlap between both continental regions. Our results, though limited in quantity, clearly demonstrate that the dynamic fingerprint microflora challenges human host inferences for forensic purposes including geographic ones. Overall, our results suggest that human fingerprint microflora is too dynamic to allow for forensic marker developments for retrieving human information.

  12. Microbial community in anaerobic hydrogen-producing microflora enriched from sludge compost.

    Science.gov (United States)

    Ueno, Y; Haruta, S; Ishii, M; Igarashi, Y

    2001-11-01

    Hydrogen production by thermophilic anaerobic microflora enriched from sludge compost was studied by using an artificial medium containing cellulose powder. Hydrogen gas was evolved with the formation of acetate, ethanol, and butyrate by decomposition of the cellulose powder. The hydrogen production yield was 2.0 mol/mol-hexose by either batch or chemostat cultivation. A medium that did not contain peptone demonstrated a lower hydrogen production yield of 1.0 mol/mol-hexose with less formation of butyrate. The microbial community in the microflora was investigated through isolation of the microorganisms by both plating and denaturing gradient gel electrophoresis (DGGE) of the' PCR-amplified V3 region of 16S rDNA. Sixty-eight microorganisms were isolated from the microflora and classified into nine distinct groups by genetic fingerprinting of the PCR-DGGE or by a random amplified polymorphic DNA analysis and determination of the partial sequence of 16S rDNA. Most of the isolates belonged to the cluster of the thermophilic Clostridium/Bacillus subphylum of low G+C gram-positive bacteria. Product formation by most of the isolated strains corresponded to that produced by the microflora. Thermoanaerobacterium thermosaccharolyticium was isolated in the enrichment culture with or without added peptone. and was detected with strong intensity by PCR-DGGE. Two other thermophilic cellulolytic microorganisms, Clostridium thermocellum and Clostridium cellulosi, were also detected by PCR-DGGE, although they could not be isolated. These findings imply that hydrogen production from cellulose by microflora is performed by a consortium of several species of microorganisms.

  13. Salinity alters snakeskin and mesh transcript abundance and permeability in midgut and Malpighian tubules of larval mosquito, Aedes aegypti.

    Science.gov (United States)

    Jonusaite, Sima; Donini, Andrew; Kelly, Scott P

    2017-03-01

    This study examined the distribution and localization of the septate junction (SJ) proteins snakeskin (Ssk) and mesh in osmoregulatory organs of larval mosquito (Aedes aegypti), as well as their response to altered environmental salt levels. Ssk and mesh transcripts and immunoreactivity were detected in tissues of endodermal origin such as the midgut and Malpighian tubules of A. aegypti larvae, but not in ectodermally derived hindgut and anal papillae. Immunolocalization of Ssk and mesh in the midgut and Malpighian tubules indicated that both proteins are concentrated at regions of cell-cell contact between epithelial cells. Transcript abundance of ssk and mesh was higher in the midgut and Malpighian tubules of brackish water (BW, 30% SW) reared A. aegypti larvae when compared with freshwater (FW) reared animals. Therefore, [(3)H]polyethylene glycol (MW 400Da, PEG-400) flux was examined across isolated midgut and Malpighian tubule preparations as a measure of their paracellular permeability. It was found that PEG-400 flux was greater across the midgut of BW versus FW larvae while the Malpighian tubules of BW-reared larvae had reduced PEG-400 permeability in conjunction with increased Cl(-) secretion compared to FW animals. Taken together, data suggest that Ssk and mesh are found in smooth SJs (sSJs) of larval A. aegypti and that their abundance alters in association with changes in epithelial permeability when larvae reside in water of differing salt content. This latter observation suggests that Ssk and mesh play a role in the homeostatic control of salt and water balance in larval A. aegypti.

  14. Human IGF1 regulates midgut oxidative stress and epithelial homeostasis to balance lifespan and Plasmodium falciparum resistance in Anopheles stephensi.

    Directory of Open Access Journals (Sweden)

    Anna L Drexler

    2014-06-01

    Full Text Available Insulin and insulin-like growth factor signaling (IIS regulates cell death, repair, autophagy, and renewal in response to stress, damage, and pathogen challenge. Therefore, IIS is fundamental to lifespan and disease resistance. Previously, we showed that insulin-like growth factor 1 (IGF1 within a physiologically relevant range (0.013-0.13 µM in human blood reduced development of the human parasite Plasmodium falciparum in the Indian malaria mosquito Anopheles stephensi. Low IGF1 (0.013 µM induced FOXO and p70S6K activation in the midgut and extended mosquito lifespan, whereas high IGF1 (0.13 µM did not. In this study the physiological effects of low and high IGF1 were examined in detail to infer mechanisms for their dichotomous effects on mosquito resistance and lifespan. Following ingestion, low IGF1 induced phosphorylation of midgut c-Jun-N-terminal kinase (JNK, a critical regulator of epithelial homeostasis, but high IGF1 did not. Low and high IGF1 induced midgut mitochondrial reactive oxygen species (ROS synthesis and nitric oxide (NO synthase gene expression, responses which were necessary and sufficient to mediate IGF1 inhibition of P. falciparum development. However, increased ROS and apoptosis-associated caspase-3 activity returned to baseline levels following low IGF1 treatment, but were sustained with high IGF1 treatment and accompanied by aberrant expression of biomarkers for mitophagy, stem cell division and proliferation. Low IGF1-induced ROS are likely moderated by JNK-induced epithelial cytoprotection as well as p70S6K-mediated growth and inhibition of apoptosis over the lifetime of A. stephensi to facilitate midgut homeostasis and enhanced survivorship. Hence, mitochondrial integrity and homeostasis in the midgut, a key signaling center for IIS, can be targeted to coordinately optimize mosquito fitness and anti-pathogen resistance for improved control strategies for malaria and other vector-borne diseases.

  15. A Kazal-type inhibitor is modulated by Trypanosoma cruzi to control microbiota inside the anterior midgut of Rhodnius prolixus.

    Science.gov (United States)

    Soares, Tatiane S; Buarque, Diego S; Queiroz, Bruna R; Gomes, Cícera M; Braz, Glória R C; Araújo, Ricardo N; Pereira, Marcos H; Guarneri, Alessandra A; Tanaka, Aparecida S

    2015-05-01

    The triatomine insect, Rhodnius prolixus, is a vector of Trypanosoma cruzi, a protozoan parasite that causes Chagas disease. The parasite must overcome immune response and microbiota to develop inside the midgut of triatomines. In this study, we expressed, purified and characterized a Kazal-type inhibitor from the midgut of R. prolixus, named RpTI, which may be involved in microbiota - T. cruzi interactions. The qPCR showed that the RpTI transcript was primarily expressed in tissues from the intestinal tract and that it was upregulated in the anterior midgut after T. cruzi infection. A 315-bp cDNA fragment encoding the mature protein was cloned into the pPIC9 vector and expressed in Pichia pastoris system. Recombinant RpTI (rRpTI) was purified on a trypsin-Sepharose column and had a molecular mass of 11.5 kDa as determined by SDS-PAGE analysis. This protein inhibited trypsin (Ki = 0.42 nM), whereas serine proteases from the coagulation cascade were not inhibited. Moreover, trypanocidal assays revealed that rRpTI did not interfere in the viability of T. cruzi trypomastigotes. The RpTI transcript was also knocked down by RNA interference prior to infection of R. prolixus with T. cruzi. The amount of T. cruzi in the anterior midgut was significantly lower in RpTI knockdown insects compared to the non-silenced groups. We also verified that the bacterial load is higher in the anterior midgut of silenced and infected R. prolixus compared to non-silenced and infected insects. Our results suggest that T. cruzi infection increases the expression of RpTI to mediate microbiota modulation and is important for parasite immediately after infection with R. prolixus.

  16. Netrins and Frazzled/DCC promote the migration and mesenchymal to epithelial transition of Drosophila midgut cells

    Directory of Open Access Journals (Sweden)

    Melissa Pert

    2015-01-01

    Full Text Available Mesenchymal-epithelial transitions (METs are important in both development and the growth of secondary tumours. Although the molecular basis for epithelial polarity is well studied, less is known about the cues that induce MET. Here we show that Netrins, well known as chemotropic guidance factors, provide a basal polarising cue during the Drosophila midgut MET. Both netrinA and netrinB are expressed in the visceral mesoderm, the substrate upon which midgut cells migrate, while their receptor frazzled (fra is expressed in midgut cells. Netrins are required to polarise Fra to the basal surface, and Netrins and Fra undergo mutually-dependent endocytosis, with Fra subsequently trafficking to late endosomes. Mutations to fra and netrins affect both migration and MET but to different degrees. Loss of fra strongly delays migration, midgut cells fail to extend protrusions, and apico-basal polarisation of proteins and epithelium formation is inhibited. In netrin mutants, the migration phenotype is weaker and cells still extend protrusions. However, apico-basal polarisation of proteins, including Fra, and FActin is greatly disrupted and a monolayer fails to form. Delocalised accumulations of FActin are prevalent in netrin mutants but not fra mutants suggesting delocalised Fra may disrupt the MET. βPS localisation is also affected in netrin mutants in that a basal gradient is reduced while localisation to the midgut/VM interface is increased. Since a similar effect is seen when endocytosis is inhibited, Netrin and Fra may regulate Integrin turnover. The results suggest Netrin-dependent basal polarisation of Fra is critical for the formation of an epithelium.

  17. Netrins and Frazzled/DCC promote the migration and mesenchymal to epithelial transition of Drosophila midgut cells.

    Science.gov (United States)

    Pert, Melissa; Gan, Miao; Saint, Robert; Murray, Michael J

    2015-01-23

    Mesenchymal-epithelial transitions (METs) are important in both development and the growth of secondary tumours. Although the molecular basis for epithelial polarity is well studied, less is known about the cues that induce MET. Here we show that Netrins, well known as chemotropic guidance factors, provide a basal polarising cue during the Drosophila midgut MET. Both netrinA and netrinB are expressed in the visceral mesoderm, the substrate upon which midgut cells migrate, while their receptor frazzled (fra) is expressed in midgut cells. Netrins are required to polarise Fra to the basal surface, and Netrins and Fra undergo mutually-dependent endocytosis, with Fra subsequently trafficking to late endosomes. Mutations to fra and netrins affect both migration and MET but to different degrees. Loss of fra strongly delays migration, midgut cells fail to extend protrusions, and apico-basal polarisation of proteins and epithelium formation is inhibited. In netrin mutants, the migration phenotype is weaker and cells still extend protrusions. However, apico-basal polarisation of proteins, including Fra, and FActin is greatly disrupted and a monolayer fails to form. Delocalised accumulations of FActin are prevalent in netrin mutants but not fra mutants suggesting delocalised Fra may disrupt the MET. βPS localisation is also affected in netrin mutants in that a basal gradient is reduced while localisation to the midgut/VM interface is increased. Since a similar effect is seen when endocytosis is inhibited, Netrin and Fra may regulate Integrin turnover. The results suggest Netrin-dependent basal polarisation of Fra is critical for the formation of an epithelium.

  18. Dengue virus serotype 2 infection alters midgut and carcass gene expression in the Asian tiger mosquito, Aedes albopictus

    Science.gov (United States)

    Hanley, Kathryn A.; Sundararajan, Anitha; Devitt, Nicholas P.; Schilkey, Faye D.; Hansen, Immo A.

    2017-01-01

    Background The Asian tiger mosquito, Aedes albopictus is currently an important vector for dengue, chikungunya and Zika virus, and its role in transmission of arthropod-borne viruses (arboviruses) may increase in the future due to its ability to colonize temperate regions. In contrast to Aedes aegypti, the dominant vector of dengue, chikungunya and Zika virus, genetic responses of Ae. albopictus upon infection with an arbovirus are not well characterized. Here we present a study of the changes in transcript expression in Ae. albopictus exposed to dengue virus serotype 2 via feeding on an artificial bloodmeal. Methodology/Principal findings We isolated midguts and midgut-free carcasses of Ae. albopictus fed on bloodmeals containing dengue virus as well as controls fed on virus-free control meals at day 1 and day 5 post-feeding. We confirmed infection of midguts from mosquitoes sampled on day 5 post-feeding via RT-PCR. RNAseq analysis revealed dynamic modulation of the expression of several putative immunity and dengue virus-responsive genes, some of whose expression was verified by qRT-PCR. For example, a serine protease gene was up-regulated in the midgut at 1 day post infection, which may potentially enhance mosquito susceptibility to dengue infection, while 14 leucine-rich repeat genes, previously shown to be involved in mosquito antiviral defenses, were down-regulated in the carcass at 5 days post infection. The number of significantly modulated genes decreased over time in midguts and increased in carcasses. Conclusion/Significance Dengue virus exposure results in the modulation of genes in a time- and site-specific manner. Previous literature on the interaction between mosquitoes and mosquito-borne pathogens suggests that most of the changes that occurred in Ae. albopictus exposed to DENV would favor virus infection. Many genes identified in this study warrant further characterization to understand their role in viral manipulation of and antiviral response of

  19. The 3D structure and function of digestive cathepsin L-like proteinases of Tenebrio molitor larval midgut.

    Science.gov (United States)

    Beton, Daniela; Guzzo, Cristiane R; Ribeiro, Alberto F; Farah, Chuck S; Terra, Walter R

    2012-09-01

    Cathepsin L-like proteinases (CAL) are major digestive proteinases in the beetle Tenebrio molitor. Procathepsin Ls 2 (pCAL2) and 3 (pCAL3) were expressed as recombinant proteins in Escherichia coli, purified and activated under acidic conditions. Immunoblot analyses of different T. molitor larval tissues demonstrated that a polyclonal antibody to pCAL3 recognized pCAL3 and cathepsin L 3 (CAL3) only in the anterior two-thirds of midgut tissue and midgut luminal contents of T. molitor larvae. Furthermore, immunocytolocalization data indicated that pCAL3 occurs in secretory vesicles and microvilli in anterior midgut. Therefore CAL3, like cathepsin L 2 (CAL2), is a digestive enzyme secreted by T. molitor anterior midgut. CAL3 hydrolyses Z-FR-MCA and Z-RR-MCA (typical cathepsin substrates), whereas CAL2 hydrolyses only Z-FR-MCA. Active site mutants (pCAL2C25S and pCAL3C26S) were constructed by replacing the catalytic cysteine with serine to prevent autocatalytic processing. Recombinant pCAL2 and pCAL3 mutants (pCAL2C25S and pCAL3C26S) were prepared, crystallized and their 3D structures determined at 1.85 and 2.1 Å, respectively. While the overall structure of these enzymes is similar to other members of the papain superfamily, structural differences in the S2 subsite explain their substrate specificities. The data also supported models for CAL trafficking to lysosomes and to secretory vesicles to be discharged into midgut contents.

  20. Dry heat effects on survival of indigenous soil particle microflora and particle viability studies of Kennedy Space Center soil

    Science.gov (United States)

    Ruschmeyer, O. R.; Pflug, I. J.; Gove, R.; Heisserer, Y.

    1975-01-01

    Research efforts were concentrated on attempts to obtain data concerning the dry heat resistance of particle microflora in Kennedy Space Center soil samples. The in situ dry heat resistance profiles at selected temperatures for the aggregate microflora on soil particles of certain size ranges were determined. Viability profiles of older soil samples were compared with more recently stored soil samples. The effect of increased particle numbers on viability profiles after dry heat treatment was investigated. These soil particle viability data for various temperatures and times provide information on the soil microflora response to heat treatment and are useful in making selections for spacecraft sterilization cycles.

  1. Absence of a specific subgingival microflora in adults with Down's syndrome

    NARCIS (Netherlands)

    Reuland-Bosma, W; van der Reijden, WA; van Winkelhoff, AJ

    2001-01-01

    Background: Periodontal disease in Down's syndrome (DS) is generally characterized by a high degree of bone loss. Bone loss of 5 mm or more is observed in 70% of these subjects. Among DS subjects, considerable differences in disease progression occur. So far, no studies have been conducted in which

  2. ATP Binding Cassette Transporter Mediates Both Heme and Pesticide Detoxification in Tick Midgut Cells.

    Directory of Open Access Journals (Sweden)

    Flavio Alves Lara

    Full Text Available In ticks, the digestion of blood occurs intracellularly and proteolytic digestion of hemoglobin takes place in a dedicated type of lysosome, the digest vesicle, followed by transfer of the heme moiety of hemoglobin to a specialized organelle that accumulates large heme aggregates, called hemosomes. In the present work, we studied the uptake of fluorescent metalloporphyrins, used as heme analogs, and amitraz, one of the most regularly used acaricides to control cattle tick infestations, by Rhipicephalus (Boophilus microplus midgut cells. Both compounds were taken up by midgut cells in vitro and accumulated inside the hemosomes. Transport of both molecules was sensitive to cyclosporine A (CsA, a well-known inhibitor of ATP binding cassette (ABC transporters. Rhodamine 123, a fluorescent probe that is also a recognized ABC substrate, was similarly directed to the hemosome in a CsA-sensitive manner. Using an antibody against conserved domain of PgP-1-type ABC transporter, we were able to immunolocalize PgP-1 in the digest vesicle membranes. Comparison between two R. microplus strains that were resistant and susceptible to amitraz revealed that the resistant strain detoxified both amitraz and Sn-Pp IX more efficiently than the susceptible strain, a process that was also sensitive to CsA. A transcript containing an ABC transporter signature exhibited 2.5-fold increased expression in the amitraz-resistant strain when compared with the susceptible strain. RNAi-induced down-regulation of this ABC transporter led to the accumulation of metalloporphyrin in the digestive vacuole, interrupting heme traffic to the hemosome. This evidence further confirms that this transcript codes for a heme transporter. This is the first report of heme transport in a blood-feeding organism. While the primary physiological function of the hemosome is to detoxify heme and attenuate its toxicity, we suggest that the use of this acaricide detoxification pathway by ticks may

  3. A hypothetical model of crossing Bombyx mori nucleopolyhedrovirus through its host midgut physical barrier.

    Directory of Open Access Journals (Sweden)

    Yang Cheng

    Full Text Available Bombyx mori nucleopolyhedrovirus (BmNPV is a primary pathogen of silkworm (B. mori that causes severe economic losses each year. However, the molecular mechanisms of silkworm-BmNPV interactions, especially the silkworm proteins that can interact with the virus, are still largely unknown. In this study, the total and membrane proteins of silkworm midguts were displayed using one- and two-dimensional electrophoresis. A virus overlay assay was used to detect B. mori proteins that specifically bind to BmNPV particles. Twelve proteins were located and identified using mass spectrometry, and the different expression of the corresponding genes in BmNPV susceptible and resistant silkworm strains also indicated their involvement in BmNPV infection. The 12 proteins are grouped based on their potential roles in viral infection, for example, endocytosis, intracellular transportation, and host responses. Based on these results, we hypothesize the following: I vacuolar ATP synthase catalytic subunit A and subunit B may be implicated in the process of the membrane fusion of virus and the release of the nucleocapsid into cytoplasm; II actin, enolase and phosphoglycerate kinase are cytoskeleton associated proteins and may play an important role in BmNPV intracellular transportation; III mitochondrial prohibitin complex protein 2, ganglioside-induced differentiation-associated protein, calreticulin, regucalcin-like isoform X1 and 60 kDa heat shock protein are involved in cell apoptosis regulation during BmNPV infection in larvae midguts; IV ribosomal P0 may be associated with BmNPV infection by regulating gene expression of BmNPV; V arginine kinase has a role in the antiviral activities against BmNPV. Our work should prove informative by providing multiple protein targets and a novel direction to investigate the molecular mechanisms of the interactions between silkworms and BmNPV.

  4. Mosquito cell line glycoproteins: an unsuitable model system for the Plasmodium ookinete-mosquito midgut interaction?

    Directory of Open Access Journals (Sweden)

    Wilkins Simon

    2010-03-01

    Full Text Available Abstract Background Mosquito midgut glycoproteins may act as key recognition sites for the invading malarial ookinete. Effective transmission blocking strategies require the identification of novel target molecules. We have partially characterised the surface glycoproteins of two cell lines from two mosquito species; Anopheles stephensi and Anopheles gambiae, and investigated the binding of Plasmodium berghei ookinetes to carbohydrate ligands on the cells. Cell line extracts were run on SDS-PAGE gels and carbohydrate moieties determined by blotting against a range of biotinylated lectins. In addition, specific glycosidases were used to cleave the oligosaccharides. Results An. stephensi 43 and An. gambiae 55 cell line glycoproteins expressed oligosaccharides containing oligomannose and hybrid oligosaccharides, with and without α1-6 core fucosylation; N-linked oligosaccharides with terminal Galβ1-3GalNAc or GalNAcβ1-3Gal; O-linked α/βGalNAc. An. stephensi 43 cell line glycoproteins also expressed N-linked Galβ1-4R and O-linked Galβ1-3GalNAc. Although P. berghei ookinetes bound to both mosquito cell lines, binding could not be inhibited by GlcNAc, GalNAc or Galactose. Conclusions Anopheline cell lines displayed a limited range of oligosaccharides. Differences between the glycosylation patterns of the cell lines and mosquito midgut epithelial cells could be a factor why ookinetes did not bind in a carbohydrate inhibitable manner. Anopheline cell lines are not suitable as a potential model system for carbohydrate-mediated adhesion of Plasmodium ookinetes.

  5. Evaluation of in vitro and in vivo effects of semipurified proteinase inhibitors from Theobroma seeds on midgut protease activity of Lepidopteran pest insects.

    Science.gov (United States)

    Paulillo, Luis Cesar Maffei Sartini; Sebbenn, Alexandre Magno; de Carvalho Derbyshire, Maria Tereza Vitral; Góes-Neto, Aristóteles; de Paula Brotto, Marco Aurélio; Figueira, Antonio

    2012-09-01

    We have characterized in vitro and in vivo effects of trypsin inhibitors from Theobroma seeds on the activity of trypsin- and chymotrypsin-like proteins from Lepidopteran pest insects. The action of semipurified trypsin inhibitors from Theobroma was evaluated by the inhibition of bovine trypsin and chymotrypsin activities determined by the hydrolysis of N-Benzoyl-DL-Arginine-p-Nitroanilide (BAPA) and N-Succinyl-Ala-Ala-Pho-Phe p-Nitroanilide (S-(Ala)2ProPhe-pNA). Proteinase inhibitor activities from Theobroma cacao and T. obovatum seeds were the most effective in inhibiting trypsin-like proteins, whereas those from T. obovatum and T. sylvestre were the most efficient against chymotrypsin-like proteins. All larvae midgut extracts showed trypsin-like proteolytic activities, and the putative trypsin inhibitors from Theobroma seeds significantly inhibited purified bovine trypsin. With respect to the influence of Theobroma trypsin inhibitors on intact insects, the inclusion of T. cacao extracts in artificial diets of velvet bean caterpillars (Anticarsia gemmatalis) and sugarcane borer (Diatraea saccharalis) produced a significant increase in the percentage of adult deformation, which is directly related to both the survival rate of the insects and oviposition.

  6. Investigation of the early intestinal microflora in premature infants with/without necrotizing enterocolitis using two different methods

    DEFF Research Database (Denmark)

    Smith, Birgitte; Bodé, Susan; Skov, Thomas;

    2012-01-01

    The pathophysiology of necrotizing enterocolitis (NEC) is multifactorial, and gastrointestinal bacteria are thought to play an important role. In this study, the role of microflora in the gastrointestinal tract of neonates with NEC was assessed by comparing cases with controls....

  7. Transitions in Oral and Intestinal Microflora Composition and Innate Immune Receptor-Dependent Stimulation during Mouse Development▿ †

    OpenAIRE

    Hasegawa, Mizuho; Osaka, Toshifumi; Tawaratsumida, Kazuki; Yamazaki, Takashi; Tada, Hiroyuki; Chen, Grace Y; Tsuneda, Satoshi; Núñez, Gabriel; Inohara, Naohiro

    2009-01-01

    Commensal bacteria possess immunostimulatory activities that can modulate host responses to affect development and homeostasis in the intestine. However, how different populations of resident bacteria stimulate the immune system remains largely unknown. We characterized here the ability of intestinal and oral microflora to stimulate individual pattern recognition receptors (PRRs) in bone marrow-derived macrophages and mesothelial cells. The intestinal but not oral microflora elicited age- and...

  8. Investigations into rhizosphere microflora. IV. Fungal association in different root regions of some rainy-season crops

    Directory of Open Access Journals (Sweden)

    V. B. Srivastava

    2015-05-01

    Full Text Available Non-rhizosphere, rhizosphere and rhizoplane microflora of the crown and distal regions of Echinochloa crusgalli (L. Beauv. and Paspalum scrobiculatum L. were studied from seedling stage to the harvest. The variation in bacterial and fungal flora in relation to host species, stage of development and żonę of the rhizosphere were studied. The differences between fungal and bacterial flora are described. The relation between rhizosphere microflora and roots exudates is described.

  9. Thorsellia anophelis is the dominant bacterium in a Kenyan population of adult Anopheles gambiae mosquitoes.

    Science.gov (United States)

    Briones, Aurelio M; Shililu, Josephat; Githure, John; Novak, Robert; Raskin, Lutgarde

    2008-01-01

    Anopheles gambiae mosquitoes are not known to harbor endosymbiotic bacteria. Here we show, using nucleic acid-based methods, that 16S rRNA gene sequences specific to a recently described mosquito midgut bacterium, Thorsellia anophelis, is predominant in the midgut of adult An. gambiae s.l. mosquitoes captured in residences in central Kenya, and also occurs in the aquatic rice paddy environment nearby. PCR consistently detected T. anophelis in the surface microlayer of rice paddies, which is also consistent with the surface-feeding behavior of A. gambiae s.l. larvae. Phylogenetic analysis of cloned environmental 16S rRNA genes identified four major Thorsellia lineages, which are closely affiliated to an insect endosymbiont of the genus Arsenophonus. Physiological characterizations support the hypothesis that T. anophelis is well adapted to the female anopheline midgut by utilizing blood and tolerating the alkaline conditions in this environment. The results suggest that aquatically derived bacteria such as T. anophelis can persist through mosquito metamorphosis and become well-established in the adult mosquito midgut.

  10. Transitions in oral and intestinal microflora composition and innate immune receptor-dependent stimulation during mouse development.

    Science.gov (United States)

    Hasegawa, Mizuho; Osaka, Toshifumi; Tawaratsumida, Kazuki; Yamazaki, Takashi; Tada, Hiroyuki; Chen, Grace Y; Tsuneda, Satoshi; Núñez, Gabriel; Inohara, Naohiro

    2010-02-01

    Commensal bacteria possess immunostimulatory activities that can modulate host responses to affect development and homeostasis in the intestine. However, how different populations of resident bacteria stimulate the immune system remains largely unknown. We characterized here the ability of intestinal and oral microflora to stimulate individual pattern recognition receptors (PRRs) in bone marrow-derived macrophages and mesothelial cells. The intestinal but not oral microflora elicited age- and cell type-specific immunostimulation. The immunostimulatory activity of the intestinal microflora varied among individual mice but was largely mediated via Toll-like receptor 4 (TLR4) during breast-feeding, whereas it became TLR4 independent after weaning. This transition was associated with a change from a microflora rich in TLR4-stimulatory proteobacteria to one dominated by Bacteroidales and/or Clostridiales that poorly stimulate TLR4. The major stimulatory activity of the intestinal microflora was still intact in NOD1-, NOD2-, TLR2-, TLR4-, TLR5-, TLR9-, TLR11-, ASC-, or RICK-deficient cells but still relied on the adaptor MyD88. These studies demonstrate a transition in the intestinal microflora accompanied by a dynamic change of its ability to stimulate different PRRs which control intestinal homeostasis.

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

    OpenAIRE

    Chakroun, Maissa; Ferré, Juan

    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 a...

  12. Functional characterization of the antibiotic resistance reservoir in the human microflora

    DEFF Research Database (Denmark)

    Sommer, Morten; Church, George M; Dantas, Gautam

    2010-01-01

    The increasing levels of multi-drug resistance in human pathogenic bacteria are compromising our ability to treat infectious disease. Since antibiotic resistance determinants are readily exchanged between bacteria through lateral gene transfer, there is an increasing interest in investigating...... reservoirs of antibiotic resistance accessible to pathogens. Due to the high likelihood of contact and genetic exchange with pathogens during disease progression, the human microflora warrants special attention as perhaps the most accessible reservoir of resistance genes. Indeed, numerous previous studies...... have demonstrated substantial antibiotic resistance in cultured isolates from the human microflora. By applying metagenomic functional selections, we recently demonstrated that the functional repertoire of resistance genes in the human microbiome is much more diverse than suggested using previous...

  13. Oral microflora and selection of resistance after a single dose of amoxicillin.

    Science.gov (United States)

    Khalil, D; Hultin, M; Rashid, M U; Lund, B

    2016-11-01

    The study aimed to determine the effects of a single-dose antibiotic prophylaxis on normal oral microflora. A single dose of 2 g amoxicillin was given to 29 healthy volunteers. Saliva was collected before antibiotic administration (day 1), and again on days 2, 5, 10, 17 and 24 and subjected to culturing and antibiotic sensitivity analysis. Twenty-one per cent (6/29) of the individuals carried penicillin-V- and amoxicillin-resistant viridans streptococci before antibiotic administration. After a single dose of amoxicillin there was a significant reduction in Streptococcus salivarius on days 2 and 5, a significant reduction in other viridans streptococci on day 2 and the proportion of viridans streptococci with reduced susceptibility to amoxicillin was significantly increased on days 2 and 5. A single dose of amoxicillin can cause an ecological disturbance and induce selection of resistant strains in the oral microflora.

  14. Water soluble nanocurcumin extracted from turmeric challenging the microflora from human oral cavity.

    Science.gov (United States)

    Gopal, Judy; Muthu, Manikandan; Chun, Se-Chul

    2016-11-15

    Water soluble nanocurcumin prepared from commercial turmeric powders was compared against ethanol extracted curcumin particles. The oral microflora from five different human volunteers was collected and the efficacy of solvent extracted curcumin versus water extracted nanocurcumin was demonstrated. Nanocurcumin activity against oral microflora confirms its antimicrobial potency. Confocal laser scanning microscopic results revealed the enhanced entry of nanocurcumin particles into microbial cells. The nanosized nature of nanocurcumin appears to have led to increased cellular interaction and thereby efficient destruction of microbial cells in the mouth. In addition, solubility of nanocurcumin is also believed to be a crucial factor behind its successful antimicrobial activity. This study proves that the bioactivity of a compound is greatly influenced by its solubility in water. This work recommends the use of water soluble nanocurcumin (extracted from turmeric) as potent substitute for curcumin in dental formulations.

  15. Findings of bacterial microflora in piglets infected with conventional swine plague

    Directory of Open Access Journals (Sweden)

    Prodanov Jasna

    2002-01-01

    Full Text Available Piglets infected with the conventional swine plague virus as a result of secondary bacterial infections sometimes show an insufficiently clear clinical and pathoanatomical picture, which is why the very procedure of diagnosis is complex and the final diagnosis unreliable. That is why these investigations were aimed at examining the presence of bacterial microflora in diseased and dead pilgets which were found to have the viral antigen for CSP using the fluorescent antibody technique, in cases where the pathomorphological finding was not characteristic for conventional swine plague. Autopsies of dead piglets most often showed changes in the digestive tract and lungs, with resulting technopathy and diseases of infective nature. Such findings on knowledge of a present bacterial microflora are especially important in cases when conventional swine plague is controlled on farms and an announcement that the disease has been contained is in the offing.

  16. A Lectin from Dioclea violacea Interacts with Midgut Surface of Lutzomyia migonei, Unlike Its Homologues, Cratylia floribunda Lectin and Canavalia gladiata Lectin

    Science.gov (United States)

    Monteiro Tínel, Juliana Montezuma Barbosa; Benevides, Melina Fechine Costa; Frutuoso, Mércia Sindeaux; Rocha, Camila Farias; Arruda, Francisco Vassiliepe Sousa; Vasconcelos, Mayron Alves; Pereira-Junior, Francisco Nascimento; Cajazeiras, João Batista; do Nascimento, Kyria Santiago; Martins, Jorge Luiz; Teixeira, Edson Holanda; Cavada, Benildo Sousa; dos Santos, Ricardo Pires; Lima Pompeu, Margarida Maria

    2014-01-01

    Leishmaniasis is a vector-borne disease transmitted by phlebotomine sand fly. Susceptibility and refractoriness to Leishmania depend on the outcome of multiple interactions that take place within the sand fly gut. Promastigote attachment to sand fly midgut epithelium is essential to avoid being excreted together with the digested blood meal. Promastigote and gut sand fly surface glycans are important ligands in this attachment. The purpose of the present study was to evaluate the interaction of three lectins isolated from leguminous seeds (Diocleinae subtribe), D-glucose and D-mannose-binding, with glycans on Lutzomyia migonei midgut. To study this interaction the lectins were labeled with FITC and a fluorescence assay was performed. The results showed that only Dioclea violacea lectin (DVL) was able to interact with midgut glycans, unlike Cratylia floribunda lectin (CFL) and Canavalia gladiata lectin (CGL). Furthermore, when DVL was blocked with D-mannose the interaction was inhibited. Differences of spatial arrangement of residues and volume of carbohydrate recognition domain (CRD) may be the cause of the fine specificity of DVL for glycans in the surface on Lu. migonei midgut. The findings in this study showed the presence of glycans in the midgut with glucose/mannose residues in its composition and these residues may be important in interaction between Lu. migonei midgut and Leishmania. PMID:25431778

  17. Physico-chemical properties and the microflora of Moroccan black table olives

    Directory of Open Access Journals (Sweden)

    Asehraou, A.

    1992-06-01

    Full Text Available Eleven samples of Moroccan black table olives were analyzed for their microflora and the physico-chemical factors. A survey of the most frequent microorganisms including standard plate count (SPC, counts of the indicator microorganisms (total and fecal coliforms and enterococci, staphylococci. Salmonella, sporeforming bacteria (Clostridium and Bacillus and yeasts and molds was determined. The physico-chemical factors grouped pH, aw, NaCl concentration and the ADV (Acid degree value of the fatty matter. Results reported hereby showed a low microbial load except for the yeasts and molds. The values found for the physico-chemical factors (low for the pH and the aw and high for the ADV and salt content would suggest an inhibitory effect on the initial microflora charge of the black table olives.

    Se estudiaron la microflora y las propiedades físico-químicas de aceitunas negras de mesa en 11 muestras. Los microorganismos más abundantes contados incluyeron: flora mesofila total, indicadores de contaminación fecal (Coliformes totales y fecales, Enterococcus, Staphylococcus, Salmonella, esporulados (Bacillus y Clostridium y levaduras y mohos. Los análisis físico-químicos incluyeron: aw (actividad de agua, pH, índice de acidez de la materia grasa y concentración de NaCl. Los resultados nos demuestran perfiles microbianos bajos para todos los microorganismos excepto levaduras y mohos. Los valores bajos de pH y de aw, y otros elevados del índice de acidez y del contenido en sal, nos sugiere una Inhibición de estos factores sobre e! crecimiento y/o la supervivencia de la microflora de las aceitunas negras de mesa.

  18. STUDY OF UROGENITAL TRACT MICROFLORA OF DNEPROPETROVSK FEMALES BY POLYMERASE CHAIN REACTION

    Directory of Open Access Journals (Sweden)

    Honcharova S.Y.

    2015-08-01

    Full Text Available We isolated and identified the pathogens from the urogenital tract in 100 women of 26-55 years in Diagnostic Center of Dnepropetrovsk Medical Academy by polymerase chain reaction. It was found that all investigated microflora was represented by HPV of high and low cancer risk - HSV type 1+2, Ureaplasma urealyticum, Chlamydia trachomatis, Trichomonas vaginalis, Mycoplasma hominis, and Candida yeast species. The most abundant pathogens from the urogenital tract were HPV, Ureaplasma urealyticum, and Chlamydia trachomatis.

  19. Genotoxic effects of starvation and dimethoate in haemocytes and midgut gland cells of wolf spider Xerolycosa nemoralis (Lycosidae).

    Science.gov (United States)

    Wilczek, Grażyna; Mędrzak, Monika; Augustyniak, Maria; Wilczek, Piotr; Stalmach, Monika

    2016-06-01

    The aim of this study was to assess the genotoxic effects of starvation and dimethoate (organophosphate insecticide) in female and male wolf spiders Xerolycosa nemoralis (Lycosidae) exposed to the stressors under laboratory conditions. DNA damage was measured in haemocytes and midgut gland cells using the comet assay. In response to the two stressing factors, both cell types showed %TDNA, tail length (TL) and OTM values higher in males than in females. Level of DNA damage in haemocytes was greater than in midgut gland cells. In both sexes, the strongest genotoxicity was recorded at single application of dimethoate. After five-time exposure to the pesticide, genotoxic effects of a single dose were sustained in males and reduced to the control level in females. Starvation stress was well tolerated by the females, in which neither cell type was affected by DNA damage. However, in male haemocytes food deprivation induced severe DNA damage, what suggests suppression of the defence potential at prolonged starvation periods.

  20. Is abnormal vaginal microflora a risk factor for intrauterine fetal growth restriction?

    Institute of Scientific and Technical Information of China (English)

    NatalijaVedmedovska; Dace Rezeberga; GilbertG G Donders

    2015-01-01

    Objective:To conduct a literature review in search of possible preventable causes for fetal growth restriction.Methods:We performed a systematic literature search regarding abnormal vaginal microflora and fetal growth encompassing the last 27-year (starting from 1986) in PubMed, Embase, and Cochrane Central to study the evidence that abnormal vaginal microflora is may be related to diminished fetal growth or small for date birth.Results:Most of the 14 studies suggested a significant role of vaginal organisms in impaired fetal growth, unrelated to preterm birth. The neonatal outcome has shown to be largely linked to the preventable or foreseeable fetal factors, such as genetic abnormalities, but also ascending intrauterine infections. Our previous work suggested a role of vaginal organisms in adverse pregnancy outcome, not only preterm birth, but also impaired fetal growth.Conclusions:There is a need for cohort studies designed to unravel this link between abnormal microflora and FGR, in order to enable preventive actions to protect these small babies from severe damage and death by early screening and treatment.

  1. [Microflora of active ooze participating in the decomposition of sulfanilic acid].

    Science.gov (United States)

    Orshanskaia, F B; Arkad'eva, A Z; Kozlova, E I

    1975-01-01

    Microflora of domestic water can be a source of active ooze adapted to sulphanilic acid. Adaptation of the microflora to sulphanilic acid at a concentration of 170-200 mg/l takes 6 to 8 days. The microflora of active ooze, immediately after adaptation, consists mainly of Pseudomonas species, Ps. denitrificans, Ps. fluorescens, Ps. striata, Ps. putida, etc., and also of Achromobacter stutzeri, Achromobacter flavum, Mycobacterium phlei, Mycobacterium mucosum, Bacillus mesentericus, Bac. cereus, saccharomyces cerevisiae, Schizosaccharomyces pombe, and Rhodotorula glutinus. The number of the species decreased as a result of long cultivation of active ooze on a minimal medium with sulphanilic acid as a sole source of carbon and nitrogen; the following strains prevailed: Ps. putida, Ps. eisenbergii, strains of Mycobacterium phlei and Flavobacterium solare. The isolated strains of Ps. putida and Ps. eisenbergii decomposed sulphanilic acid by 60.0--79.5 percent, and together with Mycobacterium phlei by 100 percent during 4 to 7 days. The ability to oxidize sulphanilic acid decreased after storage. Addition to the medium of other sources of carbon, nitrogen and vitamins did not restore the lost ability of the microorganisms to decompose sulphanilic acid.

  2. Improvement of intestinal microflora balance by polysaccharide from Physalis alkekengi var. francheti.

    Science.gov (United States)

    Li, Xinli; Yang, Guang; Zhang, Cuili; Wu, Dachang; Tang, Li; Xin, Yi

    2014-02-01

    A water‑soluble polysaccharide fraction (PPSB) was fractionated from Physalis alkekengi var. francheti and purified using DEAE‑52 Cellulose and Sephadex G‑200 chromatography. The physicochemical properties of PPSB and its molecular activities involved in the improvement of intestinal microflora balance were evaluated. PPSB (12.5‑25.0 mg/ml) was shown to promote the growth of Lactobacillus delbrueckii and to inhibit the growth of Escherichia coli. Denaturing gradient gel electrophoresis analysis identified Bacteroides, Lactobacillus, Helicobacter, Prevotella, Enterococcus, Odoribacter, Alistipes and Akkermansia as dominant organisms in the intestinal tract of mice. PPSB exhibited stimulatory effects on the growth of probiotic bacteria but inhibitory effects on the growth of pathogenic bacteria in vitro. The number of species and quantity of the Lactobacillus genus increased significantly with increasing concentrations of PPSB in vivo. Experimental results of this study suggest that PPSB may improve intestinal microflora imbalances and therefore has strong potential as a natural agent for restoring the intestinal microflora balance.

  3. Effect of gluconic acid on piglet growth performance, intestinal microflora, and intestinal wall morphology.

    Science.gov (United States)

    Biagi, G; Piva, A; Moschini, M; Vezzali, E; Roth, F X

    2006-02-01

    Gluconic acid (GA) derives from the incomplete oxidation of glucose by some Gluconobacter strains. When fed to nonruminant animals, GA is only poorly absorbed in the small intestine and is primarly fermented to butyric acid in the lower gut. This study investigated the effect of GA on in vitro growth response and metabolism of swine cecal microflora and on animal growth performance, intestinal wall morphology, and intestinal microflora. During a 24-h in vitro cecal fermentation, total gas production and maximum rate of gas production were increased by GA (linear, P acids, acetic acid, propionic acid, n-butyric acid, acetic to propionic acid ratio, and acetic + butyric to propionic acid ratio were linearly increased by GA (P acid bacteria were not affected by GA. Gluconic acid tended to increase total short-chain fatty acids in the jejunum (+174, +87, and +74% for GA at 3,000, 6,000, and 12,000 ppm, respectively; P of the model = 0.07; quadratic, P = 0.07). Morphological evaluation of intestinal mucosa from jejunum, ileum, and cecum did not show any significant differences among treatments. This study showed that feeding GA influences the composition and activity of the intestinal microflora and may improve growth performance of piglets after weaning.

  4. Gut microflora of vervet and samango monkeys in relation to diet.

    Science.gov (United States)

    Bruorton, M R; Davis, C L; Perrin, M R

    1991-02-01

    The microflora in the gastrointestinal tracts of wild vervet and samango monkeys (Cercopithecus aethiops and C. mitis, respectively) were studied, using fermentation acid analysis, electron microscopy, and culturing methods. The diets of the two species of monkey differ considerably, with that of the samango including a greater proportion of cellulose-rich leaf material, and this is reflected in the microflora. Volatile fatty acid measurements along the gut of both species showed that these end products of bacterial metabolism were concentrated in the cecum and colon. Electron microscopy indicated that morphologically similar bacteria were present in the cecum and colon of both species, but the samango possessed a distinct stomach microflora. Bacteria in the lumina of the four main regions of the gut of the monkeys (stomach, small intestine, cecum, and colon) were plated on a number of anaerobic media (Mann, Rogosa, and Sharp; clostridial basal; and complex media). The cecum and colon were found to contain higher numbers of microbes per gram (wet weight) of gut content than the stomach and small intestine. Microbial isolates were able to catabolize carboxymethyl cellulose and other polymers. This may aid the monkeys, particularly samangos, in the digestion of fibrous dietary components such as leaves.

  5. Effect of endogenic phototherapy on intestinal microflora and immunity of a man

    Science.gov (United States)

    Naumov, Sergey A.; Vovk, Sergey M.; Khlusov, Igor A.; Philippova, Galina V.; Volovodenko, Alexey V.; Naumov, Alexey V.; Goldberg, Viktor E.; Borodulina, Elena V.

    2001-05-01

    An endogenous photostimulator (APhS) seems to be a very small physiotherapeutic device allowing to irradiate the part of the gastrointestinal tract (GIT) with light of visible range especially in cases when modern endoscopic equipment cannot be applied. There was studied the effect of a photostimulator on composition of intestinal microflora and the immunologic inspection of patients with dysbacteriosis (dysbioses) of the intestine as a result of diseases of the gastrointestinal tract and after antibacterial therapy for other disturbances. The obtained results are comparable with indices of the control group. At the same time, it should be noted that stimulation of growth of natural flora is recorded in the main group of patients which inhibits the activity of conditioned pathogenic microflora. Indications and contraindications for application of the method of endogenic phototherapy using a photostimulator are defined. The statistical analysis of results of studying the immunity indices testifies to the fact that endogenic phototherapy leads to increased activity of the humoral link of the immunity in this category of patients. Thus, endogenic phototherapy seems to be an effective method of treatment of patients with disturbances of microflora composition of the gastrointestinal tract.

  6. Relationship between mucositis and changes in oral microflora during cancer chemotherapy.

    Science.gov (United States)

    Napeñas, Joel J; Brennan, Michael T; Bahrani-Mougeot, Farah K; Fox, Philip C; Lockhart, Peter B

    2007-01-01

    It is thought that the incidence and severity of cancer chemotherapy-associated mucositis is caused in part by changes in the oral bacterial microflora. This systematic review examined the role of oral bacterial microflora changes in the development of oral mucositis during chemotherapy. Thirteen prospective clinical trials were identified, involving 300 patients with 13 different cancer diagnoses. There was great variability in patient populations, bacterial sample collection methodology, and oral sample sites. No clear pattern regarding qualitative and quantitative oral flora changes emerged among these studies. The most frequent Gram-negative species isolated during chemotherapy were from the Enterobacteriaceae family, Pseudomonas sp. and E. coli. The most common Gram-positive species isolated were Staphylococcus sp. and Streptococcus sp. Five studies assessed the role of oral flora changes in the genesis of oral mucosal changes, with no consensus among them. More detailed studies are required to understand the relationship between chemotherapy, alterations in the nature and magnitude of the oral microflora, and the presence of mucositis.

  7. Microbiological Hazards Assessment of Psychrotrophic Microflora in Bovine Carcasses Slaughtered in North West Romania

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    Silviu Vida

    2016-11-01

    Full Text Available Gram negative bacteria have the greatest capacity to spoil the meat if kept under aerobic conditions, therefor the members of the genera Pseudomonas, Acinetobater, Psychrobacter and Moraxella, will form the dominant microflora. The purpose of this study was to assess the contamination level with psychrotropic microflora of bovine carcasses in order to prove how the results could be used to improve the slaughter process. The research was carried out between January-December 2015, in two slaughterhouses from North West Romania.  The research material was represented by 144 meat samples (slaughterhouse A, n=72, slaughterhouse B, n=72. Weekly, 3 samples were collected from refrigerated carcasses, and examined for total psychrotrophic counts (TPC, Enterobacteriaceae, Pseudomonas, Aeromonas and Yersinia. From statistical processing of recorded data was established that mean log TPC from the surface of refrigerated carcasses has presented different values, ranged between 3.70±0.20 log CFU/cm2 and 6.90±0.43 log CFU/cm2. Initial surface microflora of bovine carcasses was represented by germs from the following genera: Staphylococcus, Micrococcus, Aeromonas, Acinetobacter, Pseudomonas, Yersinia, Serratia, Hafnia, Proteus and Escherichia. Microbiological hazard assessment reveals the key role of psychotropic microorganisms in the spoilage of meat, if the monitoring system of the slaughtering process is not functioning properly.

  8. Enrichment and hydrogen production by marine anaerobic hydrogen-producing microflora

    Institute of Scientific and Technical Information of China (English)

    CAI JinLing; WANG GuangCe; LI YanChuan; ZHU DaLing; PAN GuangHua

    2009-01-01

    Acid,alkali,heat-shock,KNO3 and control pretreatment methods applied to anaerobic sludge were evaluated for their ability to selectively enrich the marine hydrogen-producing mixed microflora.Seawater culture medium was used as the substrate.The hydrogen yield of pretreated microflora was higher than that of the un-pretreated control (P<0.05).Among the pretreatment methods studied,heat-shock pretreatment yielded the greatest hydrogen production,which was 14.6 times that of the control.When the effect of initial pH on hydrogen production of heat-shock pretreated samples was studied,hydrogen was produced over the entire pH range (pH 4-10).The hydrogen yield peaked at initial pH 8 (79 mL/g sucrose) and then steadily decreased as the initial pH increased.Sucrose consumption was high at neutral initial pH.During the process of hydrogen production,pH decreased gradually,which indicated that the acquired microflora consisted of acidogenic bacteria.

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

    Science.gov (United States)

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

    2012-01-01

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

  10. A new antimicrobial protein from the anterior midgut of Triatoma infestans mediates Trypanosoma cruzi establishment by controlling the microbiota.

    Science.gov (United States)

    Buarque, Diego S; Gomes, Cícera M; Araújo, Ricardo N; Pereira, Marcos H; Ferreira, Roberta C; Guarneri, Alessandra A; Tanaka, Aparecida S

    2016-04-01

    The Reduviid Triatoma infestans is a vector for the protozoan Trypanosoma cruzi, the etiological agent of Chagas disease. The parasite must address the defense molecules and microbiota that colonize the anterior midgut of T. infestans. To obtain insight into T. cruzi - microbiota interactions in triatomine insects, we characterized a new antimicrobial product from the anterior midgut of T. infestans (TiAP) that may be involved in these relationships. The TiAP DNA fragment was cloned and expressed in a bacterial system, and the effect of the protein on bacteria and T. cruzi was evaluated by RNAi, qPCR and antimicrobial experiments. The number of T. cruzi in T. infestans anterior midguts was significantly lower in TiAP knockdown insects than in unsilenced groups. We also verified that the amount of bacteria in silenced T. infestans is approximately 600-fold higher than in unsilenced insects by qPCR. The 327-bp cDNA fragment that encodes mature TiAP was cloned into the pET-14b vector and expressed fused to a His-tag in Escherichia coli C43. The recombinant protein (rTiAP) was purified using an Ni-NTA column, followed by a HiTrap SP column. According to a trypanocidal assay, rTiAP did not interfere with the viability of T. cruzi trypomastigotes. Moreover, in antimicrobial experiments using E. coli and Micrococcus luteus, the protein was only bacteriostatic for Gram-negative bacteria. The data indicate that infection by T. cruzi increases the expression of TiAP to modulate the microbiota. The inhibition of microbiota growth by TiAP is important for parasite establishment in the T. infestans anterior midgut.

  11. Amount and metal composition of midgut gland metallothionein in shore crabs (Carcinus maenas) after exposure to cadmium in the food

    Energy Technology Data Exchange (ETDEWEB)

    Pedersen, Knud Ladegaard; Bach, Louise Thornhøj; Bjerregaard, Poul, E-mail: poul@biology.sdu.dk

    2014-05-01

    Highlights: • Crabs were fed with Cd in concentrations of 1.1–5.1 μg g⁻¹ food. • Metallothionein concentrations only increased at 5.1 μg g⁻¹. • Cd contents of metallothionein increased linearly with exposure. • A marked influence by the variable Cu contents on metal composition was recorded. • Digestive gland metallothionein is a poor biomarker for Cd exposure. - Abstract: Accumulation of cadmium in aquatic invertebrates may compromise human food safety and anthropogenic additions of cadmium to coastal areas cause concern. Induction of crustacean metallothionein has been suggested as a useful biomarker for contamination of the aquatic environment with cadmium. We investigated how exposure to low concentrations of cadmium in the food affects the subcellular binding of cadmium with the shore crab Carcinus maenas as model organism. Approximately 80% of the assimilated cadmium was bound in the soluble fraction of the midgut gland and of this, 82% was found in the metallothionein fraction. Metallothionein synthesis was only induced at the highest exposure level. However, the number of cadmium atoms bound per molecule of metallothionein increased linearly with exposure, from approximately 0.18 in the control group to 1.4 in a group administered food containing 5.1 μg Cd g⁻¹. We noted a marked interaction between the presence of copper and zinc in the midgut gland and the binding of cadmium. The usefulness of crustacean midgut gland metallothionein as a biomarker for cadmium exposure at modest levels was questioned since exposures at levels producing significant increases in the tissue contents of the metal did not result in elevated concentrations of metallothionein in the midgut gland.

  12. Effect of silicon on the morphology of the midgut and mandible of tomato leafminer Tuta absoluta (Lepidoptera: Gelechiidae) larvae

    OpenAIRE

    MC dos Santos; AM Resende Junqueira; VG Mendes de Sá; JC Zanúncio; JE. Serrão

    2015-01-01

    Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae) is an important insect pest causing serious losses to tomato plantations in Brazil. Some populations of T. absolute are reported to present insecticide resistance resulting in its control failure and the use of alternative control based on silicon, which is clean and sustainable, can reduce pesticide use, increasing fruit quality and protecting the environment. This study evaluated changes in the morphology of the midgut and the mandibles of ...

  13. A slot blot immunoassay for quantitative detection of Plasmodium falciparum circumsporozoite protein in mosquito midgut oocyst.

    Directory of Open Access Journals (Sweden)

    Sanjai Kumar

    Full Text Available There is still a need for sensitive and reproducible immunoassays for quantitative detection of malarial antigens in preclinical and clinical phases of vaccine development and in epidemiology and surveillance studies, particularly in the vector host. Here we report the results of sensitivity and reproducibility studies for a research-grade, quantitative enhanced chemiluminescent-based slot blot assay (ECL-SB for detection of both recombinant Plasmodium falciparum circumsporozoite protein (rPfCSP and native PfCSP from Oocysts (Pf Oocyst developing in the midguts of Anopheles stephensi mosquitoes. The ECL-SB detects as little as 1.25 pg of rPfCSP (linear range of quantitation 2.5-20 pg; R2 = 0.9505. We also find the earliest detectable expression of native PfCSP in Pf Oocyst by ECL-SB occurs on day 7 post feeding with infected blood meal. The ECL-SB was able to detect approximately as few as 0.5 day 8 Pf Oocysts (linear quantitation range 1-4, R2 = 0.9795 and determined that one Pf Oocyst expressed approximately 2.0 pg (0.5-3 pg of native PfCSP, suggesting a similar range of detection for recombinant and native forms of Pf CSP. The ECL-SB is highly reproducible; the Coefficient of Variation (CV for inter-assay variability for rPf CSP and native PfCSP were 1.74% and 1.32%, respectively. The CVs for intra-assay variability performed on three days for rPf CSP were 2.41%, 0.82% and 2% and for native Pf CSP 1.52%, 0.57%, and 1.86%, respectively. In addition, the ECL-SB was comparable to microscopy in determining the P. falciparum prevalence in mosquito populations that distinctly contained either high and low midgut Pf Oocyst burden. In whole mosquito samples, estimations of positivity for P. falciparum in the high and low burden groups were 83.3% and 23.3% by ECL-SB and 85.7% and 27.6% by microscopy. Based on its performance characteristics, ECL-SB could be valuable in vaccine development and to measure the parasite prevalence in mosquitoes

  14. The Midgut Bacterial Flora of Laboratory-Reared Hard Ticks, Haemaphysalis longicornis, Hyalomma asiaticum, and Rhipicephalus haemaphysaloides

    Institute of Scientific and Technical Information of China (English)

    LI Chun-hong; CAO Jie; ZHOU Yong-zhi; ZHANG Hou-shuang; GONG Hai-yan; ZHOU Jin-lin

    2014-01-01

    Ixodid ticks play an important role in the transmission of a variety of zoonoses of viral, bacterial and protozoan origin, and they also harbor a wealth of microorganisms. To gain more detailed insights into the potential interactions between bacterial lfora and tick-borne pathogens, we investigated the midgut bacterial lfora of laboratory-reared Haemaphysalis longicornis, Hyalomma asiaticum and Rhipicephalus haemaphysaloides. Based on morphological, biochemical, and 16S rDNA sequencing results, we identiifed 15 species belonging to 12 genera in the midgut of the three ticks. The bacterial communities were similar to those found in other studies of hematophagous arthropods. Kocuria sp. was the most frequently isolated species and its 16S rDNA gene sequence was very similar to Kocuria koreensis P31T. To our knowledge, this is the ifrst report of the bacterial lfora of tick midguts and the results show that there were many different bacterial species in each tick species. Among the most common genera, there may have been a novel species in the genus Kocuria. The results might be the ifrst step for looking for different aspects of the pathogen and tick interaction.

  15. The effect of starvation and re-feeding on mitochondrial potential in the midgut of Neocaridina davidi (Crustacea, Malacostraca)

    Science.gov (United States)

    Włodarczyk, Agnieszka; Sonakowska, Lidia; Kamińska, Karolina; Marchewka, Angelika; Wilczek, Grażyna; Wilczek, Piotr; Student, Sebastian; Rost-Roszkowska, Magdalena

    2017-01-01

    The midgut in the freshwater shrimp Neocaridina davidi (previously named N. heteropoda) (Crustacea, Malacostraca) is composed of a tube-shaped intestine and a large hepatopancreas that is formed by numerous blind-ended tubules. The precise structure and ultrastructure of these regions were presented in our previous papers, while here we focused on the ultrastructural changes that occurred in the midgut epithelial cells (D-cells in the intestine, B- and F- cells in the hepatopancreas) after long-term starvation and re-feeding. We used transmission electron microscopy, light and confocal microscopes and flow cytometry to describe all of the changes that occurred due to the stressor with special emphasis on mitochondrial alterations. A quantitative assessment of cells with depolarized mitochondria helped us to establish whether there is a relationship between starvation, re-feeding and the inactivation/activation of mitochondria. The results of our studies showed that in the freshwater shrimp N. davidi that were analyzed, long-term starvation activates the degeneration of epithelial cells at the ultrastructural level and causes an increase of cells with depolarized (non-active) mitochondria. The process of re-feeding leads to the gradual regeneration of the cytoplasm of the midgut epithelial cells; however, these changes were observed at the ultrastructural level. Additionally, re-feeding causes the regeneration of mitochondrial ultrastructure. Therefore, we can state that the increase in the number of cells with polarized mitochondria occurs slowly and does not depend on ultrastructural alterations. PMID:28282457

  16. Polyphenol-Rich Diets Exacerbate AMPK-Mediated Autophagy, Decreasing Proliferation of Mosquito Midgut Microbiota, and Extending Vector Lifespan

    Science.gov (United States)

    Nunes, Rodrigo Dutra; Ventura-Martins, Guilherme; Moretti, Débora Monteiro; Medeiros-Castro, Priscilla; Rocha-Santos, Carlucio; Daumas-Filho, Carlos Renato de Oliveira; Bittencourt-Cunha, Paula Rego Barros; Martins-Cardoso, Karina; Cudischevitch, Cecília Oliveira; Menna-Barreto, Rubem Figueiredo Sadok; Oliveira, José Henrique Maia; Gusmão, Desiely Silva; Alves Lemos, Francisco José; Alviano, Daniela Sales; Oliveira, Pedro Lagerblad; Lowenberger, Carl; Majerowicz, David; Oliveira, Ricardo Melo; Mesquita, Rafael Dias; Atella, Georgia Correa

    2016-01-01

    Background Mosquitoes feed on plant-derived fluids such as nectar and sap and are exposed to bioactive molecules found in this dietary source. However, the role of such molecules on mosquito vectorial capacity is unknown. Weather has been recognized as a major determinant of the spread of dengue, and plants under abiotic stress increase their production of polyphenols. Results Here, we show that including polyphenols in mosquito meals promoted the activation of AMP-dependent protein kinase (AMPK). AMPK positively regulated midgut autophagy leading to a decrease in bacterial proliferation and an increase in vector lifespan. Suppression of AMPK activity resulted in a 6-fold increase in midgut microbiota. Similarly, inhibition of polyphenol-induced autophagy induced an 8-fold increase in bacterial proliferation. Mosquitoes maintained on the polyphenol diet were readily infected by dengue virus. Conclusion The present findings uncover a new direct route by which exacerbation of autophagy through activation of the AMPK pathway leads to a more efficient control of mosquito midgut microbiota and increases the average mosquito lifespan. Our results suggest for the first time that the polyphenol content and availability of the surrounding vegetation may increase the population of mosquitoes prone to infection with arboviruses. PMID:27732590

  17. Larval midgut modifications associated with Bti resistance in the yellow fever mosquito using proteomic and transcriptomic approaches

    Directory of Open Access Journals (Sweden)

    Tetreau Guillaume

    2012-06-01

    Full Text Available Abstract Background Bacillus thuringiensis var. israelensis (Bti is a natural larval mosquito pathogen producing pore-forming toxins targeting the midgut of Diptera larvae. It is used worldwide for mosquito control. Resistance mechanisms of an Aedes aegypti laboratory strain selected for 30 generations with field-collected leaf litter containing Bti toxins were investigated in larval midguts at two levels: 1. gene transcription using DNA microarray and RT-qPCR and 2. differential expression of brush border membrane proteins using DIGE (Differential In Gel Electrophoresis. Results Several Bti Cry toxin receptors including alkaline phosphatases and N-aminopeptidases and toxin-binding V-ATPases exhibited altered expression levels in the resistant strain. The under-expression of putative Bti-receptors is consistent with Bt-resistance mechanisms previously described in Lepidoptera. Four soluble metalloproteinases were found under-transcribed together with a drastic decrease of metalloproteinases activity in the resistant strain, suggesting a role in resistance by decreasing the amount of activated Cry toxins in the larval midgut. Conclusions By combining transcriptomic and proteomic approaches, we detected expression changes at nearly each step of the ingestion-to-infection process, providing a short list of genes and proteins potentially involved in Bti-resistance whose implication needs to be validated. Collectively, these results open the way to further functional analyses to better characterize Bti-resistance mechanisms in mosquitoes.

  18. Induction of Manduca sexta Larvae Caspases Expression in Midgut Cells by Bacillus thuringiensis Cry1Ab Toxin

    Directory of Open Access Journals (Sweden)

    Helena Porta

    2011-01-01

    Full Text Available Bacillus thuringiensis produces crystal toxins known as Cry that are highly selective against important agricultural and human health-related insect pests. Cry proteins are pore-forming toxins that interact with specific receptors in the midgut cell membrane of susceptible larvae making pores that cause osmotic shock, leading finally to insect death. In the case of pore-forming toxins that are specific to mammalian cells, death responses at low doses may induce apoptosis or pyroptosis, depending on the cell type. The death mechanism induced by Cry toxins in insect midgut cells is poorly understood. Here, we analyze the caspases expression by RT-PCR analysis, showing that the initial response of Manduca sexta midgut cells after low dose of Cry1Ab toxin administration involves a fast and transient accumulation of caspase-1 mRNA, suggesting that pyroptosis was activated by Cry1Ab toxin as an initial response but was repressed later. In contrast, caspase-3 mRNA requires a longer period of time of toxin exposure to be activated but presents a sustained activation, suggesting that apoptosis may be a cell death mechanism induced also at low dose of toxin.

  19. Roles and regulation of autophagy and apoptosis in the remodelling of the lepidopteran midgut epithelium during metamorphosis

    Science.gov (United States)

    Romanelli, Davide; Casartelli, Morena; Cappellozza, Silvia; de Eguileor, Magda; Tettamanti, Gianluca

    2016-09-01

    We previously showed that autophagy and apoptosis occur in the removal of the lepidopteran larval midgut during metamorphosis. However, their roles in this context and the molecular pathways underlying their activation and regulation were only hypothesized. The results of the present study better clarify the timing of the activation of these two processes: autophagic and apoptotic genes are transcribed at the beginning of metamorphosis, but apoptosis intervenes after autophagy. To investigate the mechanisms that promote the activation of autophagy and apoptosis, we designed a set of experiments based on injections of 20-hydroxyecdysone (20E). Our data demonstrate that autophagy is induced at the end of the last larval stage by the 20E commitment peak, while the onset of apoptosis occurs concomitantly with the 20E metamorphic peak. By impairing autophagic flux, the midgut epithelium degenerated faster, and higher caspase activity was observed compared to controls, whereas inhibiting caspase activation caused a severe delay in epithelial degeneration. Our data demonstrate that autophagy plays a pro-survival function in the silkworm midgut during metamorphosis, while apoptosis is the major process that drives the demise of the epithelium. The evidence collected in this study seems to exclude the occurrence of autophagic cell death in this setting.

  20. Large accumulations of maize streak virus in the filter chamber and midgut cells of the leafhopper vector Cicadulina mbila.

    Science.gov (United States)

    Ammar, El-Desouky; Gargani, Daniel; Lett, Jean M; Peterschmitt, Michel

    2009-01-01

    Maize streak virus (MSV, Mastrevirus, Geminiviridae) is persistently transmitted by Cicadulina mbila, apparently without propagation in its leafhopper vector. MSV was shown earlier by quantitative PCR to accumulate in the alimentary canal of C. mbila. We examined the alimentary canals of C. mbila leafhoppers that acquired MSV from diseased plants for various acquisition access periods (AAP) by immunofluorescence confocal laser scanning microscopy (iCLSM) and by immunogold labelling transmission electron microscopy (iTEM). Following a 7-day AAP and a 7-day inoculation period (IP) on healthy seedlings, MSV was detected by iCLSM mainly in the filter chamber and anterior midgut. Using iTEM, large accumulations of MSV particles, usually enclosed in membranous vesicles, were detected only in cells of the midgut, inside and outside the filter chamber, following 14- or 30-day AAPs, and also following 7-day AAP and 7-day IP on healthy plants. No virus was detected in the control non-vector species C. chinaï. Coated pits or vesicles, typical of clathrin-mediated endocytosis, were not observed. We discuss an alternative endocytosis pathway and suggest that the MSV accumulations are stored in endosomes in the midgut epithelial cells.

  1. Roles and regulation of autophagy and apoptosis in the remodelling of the lepidopteran midgut epithelium during metamorphosis

    Science.gov (United States)

    Romanelli, Davide; Casartelli, Morena; Cappellozza, Silvia; de Eguileor, Magda; Tettamanti, Gianluca

    2016-01-01

    We previously showed that autophagy and apoptosis occur in the removal of the lepidopteran larval midgut during metamorphosis. However, their roles in this context and the molecular pathways underlying their activation and regulation were only hypothesized. The results of the present study better clarify the timing of the activation of these two processes: autophagic and apoptotic genes are transcribed at the beginning of metamorphosis, but apoptosis intervenes after autophagy. To investigate the mechanisms that promote the activation of autophagy and apoptosis, we designed a set of experiments based on injections of 20-hydroxyecdysone (20E). Our data demonstrate that autophagy is induced at the end of the last larval stage by the 20E commitment peak, while the onset of apoptosis occurs concomitantly with the 20E metamorphic peak. By impairing autophagic flux, the midgut epithelium degenerated faster, and higher caspase activity was observed compared to controls, whereas inhibiting caspase activation caused a severe delay in epithelial degeneration. Our data demonstrate that autophagy plays a pro-survival function in the silkworm midgut during metamorphosis, while apoptosis is the major process that drives the demise of the epithelium. The evidence collected in this study seems to exclude the occurrence of autophagic cell death in this setting. PMID:27609527

  2. Ferritin acts as the most abundant binding protein for snowdrop lectin in the midgut of rice brown planthoppers (Nilaparvata lugens).

    Science.gov (United States)

    Du, J; Foissac, X; Carss, A; Gatehouse, A M; Gatehouse, J A

    2000-04-01

    The mannose-specific snowdrop lectin [Galanthus nivalis agglutinin (GNA)] displays toxicity to the rice brown planthopper Nilaparvata lugens. A 26kDa GNA-binding polypeptide from N. lugens midgut was identified by lectin blotting and affinity chromatography, and characterized by N-terminal sequencing. This polypeptide is the most abundant binding protein for GNA in the N. lugens midgut. A cDNA (fersub2) encoding this protein was isolated from an N. lugens cDNA library. The deduced amino acid sequence shows significant homology to ferritin subunits from Manduca sexta and other arthropods, plants and vertebrates, and contains a putative N-glycosylation site. Native ferritin was purified from whole insects as a protein of more than 400kDa in size and characterized biochemically. Three subunits of 20, 26 and 27kDa were released from the native complex. The 26kDa subunit binds GNA, and its N-terminal sequence was identical to that of fersub2. A second cDNA (fersub1), exhibiting strong homology with dipteran ferritin, was identified as an abundant cDNA in an N. lugens midgut-specific cDNA library, and could encode the larger ferritin subunit. The fersub1 cDNA carries a stem-loop structure (iron-responsive element) upstream from the start codon, similar to structures that have been shown to play a role in the control of ferritin synthesis in other insects.

  3. Differential Midgut Attachment of Leishmania (Viannia braziliensis in the Sand Flies Lutzomyia (Nyssomyia whitmani and Lutzomyia (Nyssomyia intermedia

    Directory of Open Access Journals (Sweden)

    Rodrigo P. Soares

    2010-01-01

    Full Text Available The interaction between Leishmania and sand flies has been demonstrated in many Old and New World species. Besides the morphological differentiation from procyclic to infective metacyclic promastigotes, the parasite undergoes biochemical transformations in its major surface lipophosphoglycan (LPG. An upregulation of β-glucose residues was previously shown in the LPG repeat units from procyclic to metacyclic phase in Leishmania (Viannia braziliensis, which has not been reported in any Leishmania species. LPG has been implicated as an adhesion molecule that mediates the interaction with the midgut epithelium of the sand fly in the Subgenus Leishmania. These adaptations were explored for the first time in a species from the Subgenus Viannia, L. (V. braziliensis with its natural vectors Lutzomyia (Nyssomyia intermedia and Lutzomyia (Nyssomyia whitmani. Using two in vitro binding techniques, phosphoglycans (PGs derived from procyclic and metacyclic parasites were able to bind to the insect midgut and inhibit L. braziliensis attachment. Interestingly, L. braziliensis procyclic parasite attachment was ∼11-fold greater in the midgut of L. whitmani than in L. intermedia. The epidemiological relevance of L. whitmani as a vector of American Cutaneous Leishmaniasis (ACL in Brazil is discussed.

  4. Insect midgut α-mannosidases from family 38 and 47 with emphasis on those of Tenebrio molitor.

    Science.gov (United States)

    Moreira, Nathalia R; Cardoso, Christiane; Ribeiro, Alberto F; Ferreira, Clelia; Terra, Walter R

    2015-12-01

    α-Mannosidases are enzymes which remove non-reducing terminal residues from glycoconjugates. Data on both GH47 and GH38 (Golgi and lysosomal) enzymes are available. Data on insect midgut α-mannosidases acting in digestion are preliminary and do not include enzyme sequences. Tenebrio molitor midgut α-mannosidases were separated by chromatography into two activity peaks: a major (Man1) and a minor (Man2). An antibody generated against a synthetic peptide corresponding to a sequence of α-mannosidase fragment recognizes Man2 but not Man1. That fragment was later found to correspond to TmMan2 (GenBank access KP892646), showing that the cDNA coding for Man2 is actually TmMan2. TmMan2 codes for a mature α-mannosidase with 107.5 kDa. Purified Man2 originates after SDS-PAGE one band of about 72 kDa and another of 51 kDa, which sums 123 kDa, in agreement with gel filtration (123 kDa) data. These results suggest that Man2 is processed into peptides that remain noncovalently linked within the functional enzyme. The physical and kinetical properties of purified Man1 and Man2 are similar. They have a molecular mass of 123 kDa (gel filtration), pH optimum (5.6) and response to inhibitors like swainsonine (Man1 Ki, 68 nM; Man2 Ki, 63 nM) and deoxymannojirimycin (Man1 Ki, 0.12 mM; Man2 Ki, 0.15 mM). Their substrate specificities are a little different as Man2 hydrolyzes α-1,3 and α-1,6 bonds better than α-1,2, whereas the contrary is true for Man1. Thus, they pertain to Class II (GH38 α-mannosidases), that are catabolic α-mannosidases similar to lysosomal α-mannosidase. However, Man2, in contrast to true lysosomal α-mannosidase, is secreted (immunocytolocalization data) into the midgut contents. There, Man2 may participate in digestion of fungal cell walls, known to have α-mannosides in their outermost layer. The amount of family 38 α-mannosidase sequences found in the transcriptome (454 pyrosequencing) of the midgut of 9 insects pertaining to 5 orders is

  5. Rhynchophorus ferrugineus midgut cell line to evaluate insecticidal potency of different plant essential oils.

    Science.gov (United States)

    Rizwan-ul-Haq, Muhammad; Aljabr, Ahmed Mohammed

    2015-03-01

    Cell cultures can be a potent and strong tool to evaluate the insecticidal efficiency of natural products. Plant essential oils have long been used as the fragrance or curative products around the world which means that they are safer to be used in close proximity of humans and mammals. In this study, a midgut cell line, developed from Rhynchophorus ferrugineus (RPW-1), was used for screening essential oils from nine different plants. Assays revealed that higher cell mortality was observed at 500 ppm which reached to 86, 65, 60, 59, 56, 54, 54, 53, and 53%, whereas lowest cell mortality at 1 ppm remained at 41, 23, 20, 17, 16, 15, 14, 13, and 10%, for Azadirachta indica, Piper nigrum, Mentha spicata, Cammiphora myrrha, Elettaria cardamomum, Zingiber officinale, Curcuma longa, Schinus molle, and Rosmarinus officinalis, respectively. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) cell proliferation assay revealed the percentage of cell growth inhibition was highest at 500 ppm and remained at 48, 45, 42, 37, 34, 29, 24, 22, and 18% against A. indica, P. nigrum, M. spicata, C. myrrha, E. cardamomum, Z. officinale, C. longa, S. molle, and R. officinalis, respectively. Lowest LC50 value (7.98 ppm) was found for A. indica, whereas the highest LC50 (483.11 ppm) was against R. officinalis. Thus, in this study, essential oils of A. indica exhibited the highest levels of toxicity, whereas those from R. officinalis exhibited the lowest levels of toxicity toward RPW-1 cells.

  6. Transcriptome of the Lymantria dispar (gypsy moth larval midgut in response to infection by Bacillus thuringiensis.

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    Michael E Sparks

    Full Text Available Transcriptomic profiles of the serious lepidopteran insect pest Lymantria dispar (gypsy moth were characterized in the larval midgut in response to infection by Bacillus thuringiensis kurstaki, a biopesticide commonly used for its control. RNA-Seq approaches were used to define a set of 49,613 assembled transcript sequences, of which 838, 1,248 and 3,305 were respectively partitioned into high-, mid- and low-quality tiers on the basis of homology information. Digital gene expression profiles suggested genes differentially expressed at 24 hours post infection, and qRT-PCR analyses were performed for verification. The differentially expressed genes primarily associated with digestive function, including α-amylase, lipase and carboxypeptidase; immune response, including C-type lectin 4; developmental genes such as arylphorin; as well as a variety of binding proteins: cellular retinoic acid binding protein (lipid-binding, insulin-related peptide binding protein (protein-binding and ovary C/EBPg transcription factor (nucleic acid-binding. This is the first study conducted to specifically investigate gypsy moth response to a bacterial infection challenge using large-scale sequencing technologies, and the results highlight important genes that could be involved in biopesticide resistance development or could serve as targets for biologically-based control mechanisms of this insect pest.

  7. In vitro lipid transfer between lipoproteins and midgut-diverticula in the spider Polybetes pythagoricus.

    Science.gov (United States)

    Laino, Aldana; Cunningham, Mónica L; Heras, Horacio; Garcia, Fernando

    2011-12-01

    It has been already reported that most hemolymphatic lipids in the spider Polybetes pythagoricus are transported by HDL1 and VHDL lipoproteins. We studied in vitro the lipid transfer among midgut-diverticula (M-diverticula), and either hemolymph or purified lipoproteins as well as between hemolymphatic lipoproteins. M-diverticula and hemolymph were labeled by in vivo (14)C-palmitic acid injection. In vitro incubations were performed between M-diverticula and either hemolymph or isolated lipoproteins. Hemolymph lipid uptake was associated to HDL1 (67%) and VHDL (32%). Release from hemolymph towards M-diverticula showed the opposite trend, VHDL 75% and HDL1 45%. Isolated lipoproteins showed a similar behavior to that observed with whole hemolymph. Lipid transfer between lipoproteins showed that HDL1 transfer more (14)C-lipids to VHDL than vice versa. Only 38% FFA and 18% TAG were transferred from M-diverticula to lipoproteins, while on the contrary 75% and 73% of these lipids, respectively, were taken up from hemolymph. A similar trend was observed regarding lipoprotein phospholipids. This study supports the hypothesis that HDL1 and hemocyanin-containing VHDL are involved in the uptake and release of FFA, phospholipids and triacylglycerols in the spider P. pythagoricus. The data support a directional flow of lipids from HDL1 and VHDL suggesting a mode of lipid transport between lipoproteins and M-diverticula.

  8. Triterpene acids from apple peel inhibit lepidopteran larval midgut lipases and larval growth.

    Science.gov (United States)

    Christeller, John T; McGhie, Tony K; Poulton, Joanne; Markwick, Ngaire P

    2014-07-01

    Fruit extracts from apple, kiwifruit, feijoa, boysenberry, and blueberry were screened for the presence of lipase inhibitory compounds against lepidopteran larval midgut crude extracts. From 120 extracts, six showed significant inhibition with an extract from the peel of Malus × domestica cv. "Big Red" showing highest levels of inhibition. Because this sample was the only apple peel sample in the initial screen, a survey of peels from seven apple cultivars was undertaken and showed that, despite considerable variation, all had inhibitory activity. Successive solvent fractionation and LC-MS of cv. "Big Red" apple peel extract identified triterpene acids as the most important inhibitory compounds, of which ursolic acid and oleanolic acid were the major components and oxo- and hydroxyl-triterpene acids were minor components. When ursolic acid was incorporated into artificial diet and fed to Epiphyas postvittana Walker (Tortricidae: Lepidoptera) larvae at 0.16% w/v, a significant decrease in larval weight was observed after 21 days. This concentration of ursolic acid is less than half the concentration reported in the skin of some apple cultivars.

  9. Implication of the mosquito midgut microbiota in the defense against malaria parasites.

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    Yuemei Dong

    2009-05-01

    Full Text Available Malaria-transmitting mosquitoes are continuously exposed to microbes, including their midgut microbiota. This naturally acquired microbial flora can modulate the mosquito's vectorial capacity by inhibiting the development of Plasmodium and other human pathogens through an unknown mechanism. We have undertaken a comprehensive functional genomic approach to elucidate the molecular interplay between the bacterial co-infection and the development of the human malaria parasite Plasmodium falciparum in its natural vector Anopheles gambiae. Global transcription profiling of septic and aseptic mosquitoes identified a significant subset of immune genes that were mostly up-regulated by the mosquito's microbial flora, including several anti-Plasmodium factors. Microbe-free aseptic mosquitoes displayed an increased susceptibility to Plasmodium infection while co-feeding mosquitoes with bacteria and P. falciparum gametocytes resulted in lower than normal infection levels. Infection analyses suggest the bacteria-mediated anti-Plasmodium effect is mediated by the mosquitoes' antimicrobial immune responses, plausibly through activation of basal immunity. We show that the microbiota can modulate the anti-Plasmodium effects of some immune genes. In sum, the microbiota plays an essential role in modulating the mosquito's capacity to sustain Plasmodium infection.

  10. Genetic transformation of midgut bacteria from the red imported fire ant (Solenopsis invicta).

    Science.gov (United States)

    Medina, Freder; Li, Haiwen; Vinson, S Bradleigh; Coates, Craig J

    2009-05-01

    In our previous study we isolated 10 bacterial species from fourth-instar larval midguts of the red imported fire ant, Solenopsis invicta. Here we report the genetic transformation and reintroduction of three species (Kluyvera cryocrescens, Serratia marcescens, and isolate 38) into the fire ant host. All three species were transformed with the plasmid vector, pZeoDsRed. High expression levels of DsRed were observed and the plasmid is maintained in these bacteria at 37 degrees C in the absence of antibiotic selection for at least 9 days of subculturing. The transformed bacteria were successfully reintroduced into fire ant larvae and survived in the fire ant gut for at least 7 days. Upon pupal emergence, 7 days after reintroduction, transformed bacteria can still be isolated, however, most were passed out in the meconium. We further demonstrated that the engineered bacteria could be spread within the colony by feeding this meconium to naive larvae with the aid of worker fire ants.

  11. Proteomic Analysis of the Peritrophic Matrix from the Midgut of Third Instar Larvae,Musca domestica

    Institute of Scientific and Technical Information of China (English)

    WANG Yu; XIU Jiang Fan; CHENG Jin Zhi; LUO Man; ZHAO Peng; SHANG Xiao Li; WANG Tao; WU Jian Wei

    2016-01-01

    ObjectiveTo better comprehend the molecular structure and physiological functionof the housefly larval peritrophic matrix (PM), a mass spectrometry approach wasused to investigate the PM protein composition. MethodsThe PM was dissected from the midgut of the third instar larvae,and protein extracted from the PM wasevaluated using SDS-PAGE. A 1D-PAGE lane containing all protein bandswascut fromtop to bottom, the proteins in-geltrypsinised and analysedvia shotgun liquid chromatography- tandem mass spectrometry (LC-MS/MS). ResultsIn total, 374 proteins, with molecular weightsvarying from 8.225 kD to996.065 kD and isoelectric points ranging from 3.83 to 11.24 were successfully identified, most identified proteins were mainly related to immunity, digestion,nutrient metabolism and PM structure. Furthermore, many of these proteins were functionally associated with pattern binding, polysaccharide binding, structural constituent of peritrophic membrane and chitin binding, according to Gene Ontology annotation. ConclusionThe PM protein composition, which provides a basis for further functional investigations of the identified proteins,will be useful for understanding the housefly larval gut immune system and may help to identify potential targets and exploit new bioinsecticides.

  12. Isolation and Characterization of Midgut Lectin From Aedes aegypti (L. (Diptera: Culicidae

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    Tahany Hassan Ayaad

    2015-12-01

    Full Text Available ABSTRACT The present investigation deals with the isolation and characterization of a lectin from Aedes aegypti (Ae aegypti female mid gut extract that agglutinates various mammalian red blood cells (RBCs such as human three groups A, B, and O (RH+, mouse, rat, guinea-pig, sheep and goat erythrocytes. The highest activity of both crude and isolated mid gut lectins were detected against sheep RBCs. Using (NH42 SO4 fractionation, ion-exchange and mannose-CNBr-Sepharose 6B affinity chromatography techniques, Ae. aegypti midgut lectin (Aelec was purified to homogeneity.Isoelectric focusing (IEF and reducing SDS/PAGE revealed that the isolated mid gut lectin had isoelectric point (PI of 5.90, and subunits approximate molecular weights of 35.50 and 27.35 KDa. The hemagglutination (HA of lectins were Ca2+ - independent and heat-resistant. The sugar specificity of the purified Aelec was strongly inhibited by D (+-mannose and raffinose, followed by D (+ glucose. N-acetyl-D-manosamine and N-acetyl-D-glucosamine were moderate inhibitors. None of the lectins were inhibited by the disaccharides such as galactose, lactose, trehalose (IC50 up to 200 mM or fetuin up to 1% but the glycosubstances mucin and laminarin were strong inhibitors up to very low concentrations (0.030 - 0.003%.

  13. Effect of herbal medicine Juzentaihoto on hepatic and intestinal heat shock gene expression requires intestinal microflora in mouse

    Institute of Scientific and Technical Information of China (English)

    Miho Kato; Kenji Watanabe; Atsushi Ishige; Naoko Anjiki; Masahiro Yamamoto; Yoshifumi Irie; Mitsue Taniyama; Ryoko Kibe; Junichiro Oka; Yoshimi Benno

    2007-01-01

    AIM: To evaluate the role of intestinal microflora in the effects of multi-herbal medicine on gene expression in the gut and liver.METHODS: The multi-herbal medicine Juzentaihoto (JTX) was administered to five germ-free mice and regular mice for 2 wk. Among the results of the comprehensive gene chip analysis of the intestine and liver, we featured heat shock proteins (HSPs) 70 and 105 because their gene expression changed only in the presence of microflora. Real-time RT-PCR was performed to confirm the expression levels of these HSP genes. To determine whether JTX acts directly on the HSP genes, sodium arsenite (SA) was used to induce the heat shock proteins directly. To examine the change of the intestinal microflora with administration of JTX, the terminal restriction fragment polymorphism (T-RFLP) method was used. To identify the changed bacteria, DNA sequencing was performed.RESULTS: Heat shock protein gene expression,documented by gene chip and real-time RT-PCR, changed with the administration of JTX in the regular mice but not in the germ-free mice. JTX did not suppress the direct induction of the HSPs by SA. T-RFLP suggested that JTX decreased unculturable bacteria and increased Lactobacillus johnsoni. These data suggested that JTX changed the intestinal microflora which, in turn, changed HSP gene expression.CONCLUSION: Intestinal microflora affects multi-herbal product JTX on the gene expression in the gut and liver.

  14. In vivo binding of the Cry11Bb toxin of Bacillus thuringiensis subsp. medellin to the midgut of mosquito larvae (Diptera: Culicidae

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    Ruiz Lina María

    2004-01-01

    Full Text Available Bacillus thuringiensis subsp. medellin produces numerous proteins among which 94 kDa known as Cry11Bb, has mosquitocidal activity. The mode of action of the Cry11 proteins has been described as similar to those of the Cry1 toxins, nevertheless, the mechanism of action is still not clear. In this study we investigated the in vivo binding of the Cry11Bb toxin to the midgut of the insect species Anopheles albimanus, Aedes aegypti, and Culex quinquefasciatus by immunohistochemical analysis. Spodoptera frugiperda was included as negative control. The Cry11Bb protein was detected on the apical microvilli of the midgut epithelial cells, mostly on the posterior midgut and gastric caeca of the three mosquito species. Additionally, the toxin was detected in the Malpighian tubules of An. albimanus, Ae. aegypti, Cx. quinquefasciatus, and in the basal membrane of the epithelial cells of Ae. aegypti midgut. No toxin accumulation was observed in the peritrophic membrane of any of the mosquito species studied. These results confirm that the primary site of action of the Cry11 toxins is the apical membrane of the midgut epithelial cells of mosquito larvae.

  15. Spatial distribution of digestive proteinases in the midgut of the Pacific white shrimp (Litopenaeus vannamei) indicates the existence of endo-ectoperitrophic circulation in Crustacea.

    Science.gov (United States)

    Alexandre, Daniel; Ozório, Renata A; Derner, Roberto B; Fracalossi, Débora M; Oliveira, Gabriel B; Samuels, Richard I; Terra, Walter R; Silva, Carlos P

    2014-01-01

    The effect of dietary protein concentration on the spatial distribution of digestive proteinases in the shrimp Litopenaeus vannamei indicates the existence of endo-ectoperitrophic enzyme circulation in this species. Samples recovered from the midgut gland tissues, stomach contents, three different portions of the midgut and feces were used for quantitative and qualitative analyses of the composition and distribution of the digestive proteinases. Animals were divided into three different groups: (1) animals (controls) fed with a commercial 35% protein diet, (2) animals fed with a commercial diet supplemented with ovalbumin to a final protein concentration of 60%; (3) animals fed with an 80% protein diet. Quantitative determinations using different substrates and zymograms showed that increasing protein concentration in the diet alters the distribution of proteinases along the digestive tract. Composition of proteinases in the midgut gland, stomach contents, midgut sections and feces were similar, but not identical. Chymotrypsin and trypsin paralogues were identified in all enzyme sources in a concentration gradient along the midgut in the control shrimp, the expected distribution supporting the existence of a recycling mechanism. The occurrence of a peritrophic membrane in other Decapoda suggests that endo-ectoperitrophic circulation of digestive enzymes and nutrients may also occur in other crustaceans and also extends beyond the Insecta.

  16. Comparison between Gram stain and culture for the characterization of vaginal microflora: Definition of a distinct grade that resembles grade I microflora and revised categorization of grade I microflora

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    De Backer Ellen

    2005-10-01

    Full Text Available Abstract Background The microbiological diagnosis of bacterial vaginosis is usually made using Nugent's criteria, a useful but rather laborious scoring system based on counting bacterial cell types on Gram stained slides of vaginal smears. Ison and Hay have simplified the score system to three categories and added a fourth category for microflora with a predominance of the Streptococcus cell type. Because in the Nugent system several cell types are not taken into account for a final score, we carried out a detailed assessment of the composition of the vaginal microflora in relation to standard Gram stain in order the improve the diagnostic value of the Gram stain. To this purpose we compared Gram stain based categorization of vaginal smears with i species specific PCR for the detection of Gardnerella vaginalis and Atopobium vaginae and with ii tDNA-PCR for the identification of most cultivable species. Results A total of 515 samples were obtained from 197 pregnant women, of which 403 (78.3% were categorized as grade I microflora, 46 (8.9% as grade II, 22 (4.3% as grade III and 8 (1.6% as grade IV, according to the criteria of Ison and Hay. Another 36 samples (7.0% were assigned to the new category 'grade I-like', because of the presence of diphtheroid bacilli cell types. We found that 52.7% of the grade I-like samples contained Bifidobacterium spp. while L. crispatus was present in only 2.8% of the samples and G. vaginalis and A. vaginae were virtually absent; in addition, the species diversity of this category was similar to that of grade II specimens. Based on the presence of different Lactobacillus cell types, grade I specimens were further characterized as grade Ia (40.2%, grade Iab (14.9% and grade Ib (44.9%. We found that this classification was supported by the finding that L. crispatus was cultured from respectively 87.0% and 76.7% of grade Ia and Iab specimens while this species was present in only 13.3% of grade Ib specimens, a

  17. Urolithins, ellagic acid-derived metabolites produced by human colonic microflora, exhibit estrogenic and antiestrogenic activities.

    Science.gov (United States)

    Larrosa, Mar; González-Sarrías, Antonio; García-Conesa, María Teresa; Tomás-Barberán, Francisco A; Espín, Juan Carlos

    2006-03-08

    Urolithins A and B (hydroxy-6H-dibenzo[b,d]pyran-6-one derivatives) are colonic microflora metabolites recently proposed as biomarkers of human exposure to dietary ellagic acid derivatives. Molecular models suggest that urolithins could display estrogenic and/or antiestrogenic activity. To this purpose, both urolithins and other known phytoestrogens (genistein, daidzein, resveratrol, and enterolactone) were assayed to evaluate the capacity to induce cell proliferation on the estrogen-sensitive human breast cancer MCF-7 cells as well as the ability to bind to alpha- and beta-estrogen receptors. Both urolithins A and B showed estrogenic activity in a dose-dependent manner even at high concentrations (40 microM), without antiproliferative or toxic effects, whereas the other phytoestrogens inhibited cell proliferation at high concentrations. Overall, urolithins showed weaker estrogenic activity than the other phytoestrogens. However, both urolithins displayed slightly higher antiestrogenic activity (antagonized the growth promotion effect of 17-beta-estradiol in a dose-dependent manner) than the other phytoestrogens. The IC(50) values for the ERalpha and ERbeta binding assays were 0.4 and 0.75 microM for urolithin A; 20 and 11 microM for urolithin B; 3 and 0.02 for genistein; and 2.3 and 1 for daidzein, respectively; no binding was detected for resveratrol and enterolactone. Urolithins A and B entered into MCF-7 cells and were metabolized to yield mainly urolithin-sulfate derivatives. These results, together with previous studies regarding absorption and metabolism of dietary ellagitannins and ellagic acid in humans, suggest that the gut microflora metabolites urolithins are potential endocrine-disrupting molecules, which could resemble other described "enterophytoestrogens" (microflora-derived metabolites with estrogenic/antiestrogenic activity). Further research is warranted to evaluate the possible role of ellagitannins and ellagic acid as dietary "pro-phytoestrogens".

  18. Changes in subgingival microflora after placement and removal of fixed orthodontic appliances

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    Živković-Sandić Marija

    2014-01-01

    Full Text Available Introduction. The placement of fixed orthodontic appliances may lead to increased plaque accumulation and changes in subgingival microflora. Objective. The aim of this study was to examine the changes in frequency of subgingival microflora that occur after placement and removal of fixed orthodontic appliance using polymerase chain reaction (PCR. Methods. This study included 33 orthodontic patients, who were divided into two groups. Subgingival plaque samples were collected from the right upper incisor (U1 and right upper first molar (U6. In group A, the samples were taken three times: before placement appliance (T1, after one month (T2, and after 3 months (T3. In group B the samples were also taken three times: before appliance removal (T1, after one month (T2, and after three months (T3. PCR method was used to determine the presence of P. gingivalis, A. actinomycetemcomitans, T. forsythia, and P. intermedia. Results. In group A the frequency of P. gingivalis showed statistically significant decrease at U1 (p=0.049 and U6 (p=0.008, from T1 to T2, and at U1 (p=0.048 from T1 to T3. In group B only the frequency of T. forsythia showed a statistically significant decrease, at U6 (T1 vs. T2, p=0.004; T1 vs. T3, p=0.0003. Regarding other analyzed bacteria, changes in the presence were noticed but no statistical significance was found. Conclusion. Placement of fixed appliances may have an impact on subgingival microflora, but in the first months after the placement and removal of the appliance changes were not significant, probably due to good oral hygiene. [Projekat Ministarstva nauke Republike Srbije, br. 175075

  19. Closed gastroschisis, vanishing midgut and extreme short bowel syndrome: Case report and review of the literature.

    Science.gov (United States)

    Dennison, F A

    2016-08-01

    Gastroschisis alone has excellent survival rates. Occasionally reported is closed gastroschisis, leading to vanishing small bowel and extreme short bowel syndrome. It is believed that the abdominal wall defect can contract or close in utero, which leads to strangulation of the eviscerated bowel and the rare "vanishing gut syndrome." This has a very poor prognosis with mortality as high as 70%. An 18-year-old primigravid patient's 13 week scan diagnosed a large gastroschisis affecting the fetus. After counselling, she decided to continue with the pregnancy. Between 20 and 22 weeks, the gastroschisis disappeared, and the bowel within the abdomen became markedly dilated. Spontaneous labour occurred at 33 + 3 weeks gestation. There was no abdominal wall defect seen at delivery. Imaging and an exploratory laparotomy demonstrated absence of most of the midgut. Because available options for treatment would be very aggressive and risky, palliative care was thought to be the most feasible and practical option. He died at home on day 29 after birth. Extreme short gut syndrome (less than 25 cm of remaining small bowel) is rare. There are 13 reported cases in the literature from year 2000 to 2013. Treatment is aggressive and involves a bowel lengthening procedure or small bowel transplant. All require total parenteral nutrition and liver failure, and liver transplant is a common complication. Of these cases, 12 were born alive and 7 had aggressive treatment. Only two cases were confirmed to still be alive in infancy. If gastroschisis is seen to be reducing and "disappearing" antenatally, parents should be made aware of this rare complication so that they might be prepared if a poor outcome is anticipated.

  20. Maize toxin degrades peritrophic matrix proteins and stimulates compensatory transcriptome responses in fall armyworm midgut.

    Science.gov (United States)

    Fescemyer, Howard W; Sandoya, Germán V; Gill, Torrence A; Ozkan, Seval; Marden, James H; Luthe, Dawn S

    2013-03-01

    Understanding the molecular mechanisms underlying insect compensatory responses to plant defenses could lead to improved plant resistance to herbivores. The Mp708 inbred line of maize produces the maize insect resistant 1-cysteine protease (Mir1-CP) toxin. Reduced feeding and growth of fall armyworm larvae fed on Mp708 was previously linked to impairment of nutrient utilization and degradation of the midgut (MG) peritrophic matrix (PM) by Mir1-CP. Here we examine the biochemical and transcriptional responses of fall armyworm larvae to Mir1-CP. Insect Intestinal Mucin (IIM) was severely depleted from pure PMs treated in vitro with recombinant Mir1-CP. Larvae fed on Mp708 midwhorls excrete frass largely depleted of IIM. Cracks, fissures and increased porosity previously observed in the PM of larvae fed on Mp708 midwhorls could ensue when Mir1-CP degrades the IIM that cross-links chitin fibrils in the PM. Both targeted and global transcriptome analyses were performed to determine how complete dissolution of the structure and function of the PM is prevented, enabling larvae to continue growing in the presence of Mir1-CP. The MGs from fall armyworm fed on Mp708 upregulate expression of genes encoding proteins involved in PM production as an apparent compensation to replace the disrupted PM structure and restore appropriate counter-current MG gradients. Also, several families of digestive enzymes (endopeptidases, aminopeptidases, lipases, amylase) were more highly expressed in MGs from larvae fed on Mp708 than MGs from larvae fed on diets lacking Mir1-CP (artificial diet, midwhorls from Tx601 or B73 maize). Impaired growth of larvae fed on Mp708 probably results from metabolic costs associated with higher production of PM constituents and digestive enzymes in a compensatory attempt to maintain MG function.

  1. Depressive disorders development in connection with gut microflora and dietary factors

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    Shuldyakov А.А.

    2016-06-01

    Full Text Available The morbidity of depressive disorders and their role in the formation of different human pathologies emphasize the actuality of searching for new approaches in the prophylaxis and treatment of depressions. The modification of gut microbiota may be a perspective direction of clinical investigation. Development in this area is based on the review of trials which confirm the significance of gut microflora composition in regulation of mental functions in particular mood and behavior as well as the role of diet in the development of depressions. The hypothesis of the role of gut microbiota in the development of depressive conditions was discussed.

  2. Impact of probiotic drugs, based on Enterobacter faecium autostrains, on human intestinal microflora in confined habitat

    Science.gov (United States)

    Viacheslav, Ilyin; Batov, Alexey; Usanova, Nonna

    The aim of research: Investigation of influence of probiotic drugs based on autostrains of Enter-obacter faecium, selected from the crew in long term isolation experiment in confined habitat. It is known that during long-term presence in confined habitat the risk of infectious diseases increases. One of the main infectious risk occurs during first 20 days of isolation as a result of exchange of strains and stress-mediated disbacterioses. Therefore it is necessary to evaluate activities of probiotics to avoid this risk. Furthermore, in case of super long term autonomous flight there should be possibilities of application of autochthonous microflora strains as pro-biotics to strengthen colonial resistance of crews. Materials and methods: In the experiment there were used probiotic drugs based on autostrains of E. faecium, selected from the crew before the experiment. Probiotic drugs were consumed during 30 days since the beginning of the experiment with the break of consumption between 10th to 19th day. Results: Comparing the state of intestinal microflora of the crew on the baseline and 14th day of experiment re-vealed remarkable changes of microflora: the increasing of concentration of bifidobacteria and E. faecium (approximately 10 times), elimination of hemolytic streptococcus, yeasts, reduction of the rate of S.aureus, hemolytic gramnegative non-fermenting rods, lactobacilli and normal E.coli. On the 45th day of isolation, 15 days after finishing of auto-strains administration, there fere signs of restoration of disbacteriosis: the quantitative decreasing lactobacilli, bifidobacteria and normal E.coli, increasing of the rate of S.aureus, hemolytic gramnegative nonfermentive rods. Conclusion: Thus we managed to avoid risk of pathogenicity potential growth in first 2 decades of isolation. Application of probiotic, based on the autostrains of E. faecium leads to insignificant changes of concentration of lactobacteries, bifidobacteries, normal E. coli and to

  3. Effects of high hydrostatic pressure on microflora and some quality attributes of grape juice

    Science.gov (United States)

    Mert, Mecnun; Buzrul, Sencer; Alpas, Hami

    2013-03-01

    Red and white grape juices were treated with high hydrostatic pressure (HHP) at three different pressures, temperature and time values to investigate the effects of HHP on natural microflora and some quality attributes of the juices. Increased pressure, temperature and time showed significant effect on the microbial reduction and no microbial growth were observed in HHP-treated grape juices up to 90 days. HHP had little or no effect on pH and color of the juices. Although 5-hydroxymethylfurfural (HMF) formation was observed in heat pasteurized samples, no HMF was detected in HHP-treated juices. This study demonstrated that HHP could be used as an alternative to heat treatment.

  4. THE ANTIMICROBIAL EFFECT OF GARLIC (ALLIUM SATIVUM EXTRACT ON MOUTH MICROFLORA

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    S.Mehrabian

    1995-12-01

    Full Text Available Five strains of mouth normal microflora containing staphylococcus epidermidis, streptococcus mutant, lactobacillus acidophilus, Actinomyces viscosos and candida albicans, were isolated and identified then cultured and purified on different media by using routine methods in microbiology. Extract of 3 garlic samples south (Khuzestan, north (Mazanderan and Hamedan garlics, were affected on these microbial cultures using Bauer and Kirby method. The effects were compared with antimicrobial effects of erythromycin. The results show all three used extracts have antimicrobial effects. The extract had a bactericidal effect on bacteria but a fungi static on candida albicans.

  5. Fungal microflora biodiversity as a function of pollution in Oued Sebou (Morocco).

    Science.gov (United States)

    Sage, L; Bennasser, L; Steiman, R; Seigle-Murandi, F

    1997-08-01

    An inventory of the fungal microflora present in sediments collected at 11 sites along Oued Sebou has been established. The influence of some climatic parameters (temperature, rainfall) and of pollution on the composition of the fungal communities has been questioned. Emericella nidulans, Eurotium amstelodami, Neosartotya fisheri var. glabrum (Ascomycetes), Dreschslera biseptata (Dematiaceae), Penicillium citrinum (Mucedinaceae) and Phoma sp. (Sphaeropsidale) can be considered as strains sensitive to pollution. Penicillium janthelinum, Trichoderma koningii (Mucedinaceae) and Candida albicans (Yeast) were found only at the most polluted sites; Talaromyces flavus (Ascomycete) Rhodotorula rubra, Saccharomyces cerevisiae (Yeasts) and Fusarium oxysporum (Tuberculariales) were more often found at heavily polluted sites than at lightly polluted ones.

  6. Pglyrp-Regulated Gut Microflora Prevotella falsenii, Parabacteroides distasonis and Bacteroides eggerthii Enhance and Alistipes finegoldii Attenuates Colitis in Mice.

    Science.gov (United States)

    Dziarski, Roman; Park, Shin Yong; Kashyap, Des Raj; Dowd, Scot E; Gupta, Dipika

    2016-01-01

    Dysbiosis is a hallmark of inflammatory bowel disease (IBD), but it is unclear which specific intestinal bacteria predispose to and which protect from IBD and how they are regulated. Peptidoglycan recognition proteins (Pglyrps) are antibacterial, participate in maintaining intestinal microflora, and modulate inflammatory responses. Mice deficient in any one of the four Pglyrp genes are more sensitive to dextran sulfate sodium (DSS)-induced colitis, and stools from Pglyrp-deficient mice transferred to wild type (WT) germ-free mice predispose them to much more severe colitis than stools from WT mice. However, the identities of these Pglyrp-regulated bacteria that predispose Pglyrp-deficient mice to colitis or protect WT mice from colitis are not known. Here we identified significant changes in β-diversity of stool bacteria in Pglyrp-deficient mice compared with WT mice. The most consistent changes in microbiome in all Pglyrp-deficient mice were in Bacteroidales, from which we selected four species, two with increased abundance (Prevotella falsenii and Parabacteroides distasonis) and two with decreased abundance (Bacteroides eggerthii and Alistipes finegoldii). We then gavaged WT mice with stock type strains of these species to test the hypothesis that they predispose to or protect from DSS-induced colitis. P. falsenii, P. distasonis, and B. eggerthii all enhanced DSS-induced colitis in both WT mice with otherwise undisturbed intestinal microflora and in WT mice with antibiotic-depleted intestinal microflora. By contrast, A. finegoldii (which is the most abundant species in WT mice) attenuated DSS-induced colitis both in WT mice with otherwise undisturbed intestinal microflora and in WT mice with antibiotic-depleted intestinal microflora, similar to the colitis protective effect of the entire normal microflora. These results identify P. falsenii, P. distasonis, and B. eggerthii as colitis-promoting species and A. finegoldii as colitis-protective species.

  7. 肠道微生态与肝脏疾病%The Gut Microflora and Liver Disease

    Institute of Scientific and Technical Information of China (English)

    白文元; 刘娜; 白研

    2013-01-01

    近年来国内外基础与临床研究表明,肠道微生态与肝脏疾病有密切的关系.本文重点介绍肠道微生态与肝脏疾病的相关性、肝脏疾病时对肠道微生态产生什么影响、调整肠道微生态失衡对肝脏疾病有何作用.客观评价肝脏疾病微生态基础与临床研究结论,藉以引起大家重视两者之间的关系,指导临床合理应用微生态制剂,从而达到延长肝病患者生存期,并改善其预后.%In recent years, both domestic and international basic and clinical studies have shown that there is a close relationship between gut microflora and liver disease. This article focused on the close relationship between gut microflora and liver disease, stating what is the impact on adjustment of gut microflora by liver disease, and what is the effect of adjustment of microflora imbalance on liver disease. Objective evaluation of basic microflora and clinical research in liver disease could draw our attention to the relationship between gut microflora and liver disease, guiding the clinical application of probiotics for prolonging the survival time of patients with liver disease and improving the prognosis.

  8. The associated microflora to the larvae of human bot fly Dermatobia hominis L. Jr. (Diptera: Cuterebridae and its furuncular lesions in cattle

    Directory of Open Access Journals (Sweden)

    E Sancho

    1996-06-01

    Full Text Available The microflora associated to furuncular lesions, larvae and pupae of Dermatobia hominis, as well as the relationships between parasite, host and microflora associated, as a comprehensive microsystem, has been studied. One hundred and two furuncular myiasis due to D. hominis larvae in several breeds of cattle were studied and the following bacterial species were significant: Staphylococcus aureus, S. epidermidis, S. warneri, Bacillus subtilis and Escherichia coli. Closely related, the microflora associated to 141 samples from first, second, third instar larva and both external surface and larval cavities has been studied. The representative associated microflora to the larvae were: S. aureus, B. subtilis, S. hycus and Moraxella phenylpiruvica, Moerella wisconsiensis, Proteus mirabilis and P. vulgaris, M. phenylpiruvica, M. wisconsiensis, P. mirabilis and P. rettgeri were the representative microflora associated to 64 pupae of D. hominis.

  9. The antioxidant role of xanthurenic acid in the Aedes aegypti midgut during digestion of a blood meal.

    Directory of Open Access Journals (Sweden)

    Vitor L A Lima

    Full Text Available In the midgut of the mosquito Aedes aegypti, a vector of dengue and yellow fever, an intense release of heme and iron takes place during the digestion of a blood meal. Here, we demonstrated via chromatography, light absorption and mass spectrometry that xanthurenic acid (XA, a product of the oxidative metabolism of tryptophan, is produced in the digestive apparatus after the ingestion of a blood meal and reaches milimolar levels after 24 h, the period of maximal digestive activity. XA formation does not occur in the White Eye (WE strain, which lacks kynurenine hydroxylase and accumulates kynurenic acid. The formation of XA can be diminished by feeding the insect with 3,4-dimethoxy-N-[4-(3-nitrophenylthiazol-2-yl] benzenesulfonamide (Ro-61-8048, an inhibitor of XA biosynthesis. Moreover, XA inhibits the phospholipid oxidation induced by heme or iron. A major fraction of this antioxidant activity is due to the capacity of XA to bind both heme and iron, which occurs at a slightly alkaline pH (7.5-8.0, a condition found in the insect midgut. The midgut epithelial cells of the WE mosquito has a marked increase in occurrence of cell death, which is reversed to levels similar to the wild type mosquitoes by feeding the insects with blood supplemented with XA, confirming the protective role of this molecule. Collectively, these results suggest a new role for XA as a heme and iron chelator that provides protection as an antioxidant and may help these animals adapt to a blood feeding habit.

  10. Effects of Cylindrospermopsis raciborskii and Aphanizomenon ovalisporum (cyanobacteria) ingestion on Daphnia magna midgut and associated diverticula epithelium

    Energy Technology Data Exchange (ETDEWEB)

    Nogueira, Isabel C.G. [Centro Interdisciplinar de Investigacao Marinha e Ambiental, Rua dos Bragas 177-289, 4050-123 Porto (Portugal)]. E-mail: isabelnogueira@ciimar.up.pt; Lobo-da-Cunha, Alexandre [Centro Interdisciplinar de Investigacao Marinha e Ambiental, Rua dos Bragas 177-289, 4050-123 Porto (Portugal); Laboratorio de Biologia Celular, Instituto de Ciencias Biomedicas Abel Salazar (ICBAS), Universidade do Porto, Largo Abel Salazar 2, 4099-003 Porto (Portugal); Vasconcelos, Vitor M. [Centro Interdisciplinar de Investigacao Marinha e Ambiental, Rua dos Bragas 177-289, 4050-123 Porto (Portugal); Departamento de Zoologia e Antropologia, Faculdade de Ciencias, Universidade do Porto, Praca Gomes Teixeira, 4099-002 Porto (Portugal)

    2006-11-16

    This article reports a light and electron microscopy investigation of the effects of Cylindrospermopsis raciborskii and Aphanizomenon ovalisporum ingestion on midgut and associated digestive diverticula of Daphnia magna. Additionally, survivorship and growth effects caused by feeding on cyanobacteria were assessed. Three cyanobacteria were used in the experiments: cylindrospermopsin (CYN)-producing C. raciborskii, CYN-producing A. ovalisporum and non-CYN-producing C. raciborskii. In order to discriminate between the alterations due to the low nutritional value of cyanobacteria and toxic effects, a control group was fed on the chlorophyte Ankistrodesmus falcatus and another control group was not fed. In the chlorophyte fed control, the epithelium lining the midgut and associated diverticula is mainly formed by strongly stained cells with an apical microvilli border. Nevertheless, unstained areas in which cell lyses had occurred were also observed. In the unfed control, the unstained areas became predominant due to an increment of cell lyses. All individuals fed on CYN-producing A. ovalisporum and some of those fed on non-CYN-producing C. raciborskii appear similar to the unfed control. However, some individuals fed on non-CYN-producing C. raciborskii showed similarities with the fed control. In contrast, the midgut and digestive diverticula of D. magna fed on CYN-producing C. raciborskii showed a widespread dissociation of epithelial cells, associated with severe intracellular disorganization, but cell lysis was less evident than in controls. These alterations cannot be attributed to CYN, because those effects were not induced by CYN-producing A. ovalisporum. Therefore, data suggest the production of another unidentified active metabolite by CYN-producing C. raciborskii, responsible for the disruption of cell adhesion in the epithelium of D. magna digestive tract. Data also show that the tested cyanobacteria are inadequate as food to D. magna, due to low nutritional

  11. Stage-specific adhesion of Leishmania promastigotes to sand fly midguts assessed using an improved comparative binding assay.

    Directory of Open Access Journals (Sweden)

    Raymond Wilson

    Full Text Available BACKGROUND: The binding of Leishmania promastigotes to the midgut epithelium is regarded as an essential part of the life-cycle in the sand fly vector, enabling the parasites to persist beyond the initial blood meal phase and establish the infection. However, the precise nature of the promastigote stage(s that mediate binding is not fully understood. METHODOLOGY/PRINCIPAL FINDINGS: To address this issue we have developed an in vitro gut binding assay in which two promastigote populations are labelled with different fluorescent dyes and compete for binding to dissected sand fly midguts. Binding of procyclic, nectomonad, leptomonad and metacyclic promastigotes of Leishmania infantum and L. mexicana to the midguts of blood-fed, female Lutzomyia longipalpis was investigated. The results show that procyclic and metacyclic promastigotes do not bind to the midgut epithelium in significant numbers, whereas nectomonad and leptomonad promastigotes both bind strongly and in similar numbers. The assay was then used to compare the binding of a range of different parasite species (L. infantum, L. mexicana, L. braziliensis, L. major, L. tropica to guts dissected from various sand flies (Lu. longipalpis, Phlebotomus papatasi, P. sergenti. The results of these comparisons were in many cases in line with expectations, the natural parasite binding most effectively to its natural vector, and no examples were found where a parasite was unable to bind to its natural vector. However, there were interesting exceptions: L. major and L. tropica being able to bind to Lu. longipalpis better than L. infantum; L. braziliensis was able to bind to P. papatasi as well as L. major; and significant binding of L. major to P. sergenti and L. tropica to P. papatasi was observed. CONCLUSIONS/SIGNIFICANCE: The results demonstrate that Leishmania gut binding is strictly stage-dependent, is a property of those forms found in the middle phase of development (nectomonad and leptomonad

  12. Environmental biodegradability of diesel oil: composition and performances of degradative micro-floras; Biodegradabilite du gazole dans l'environnement: composition et performances des microflores degradatrices

    Energy Technology Data Exchange (ETDEWEB)

    Penet, S.

    2004-09-01

    The large use of petroleum products makes them a significant source of pollutants in ground water and soils. Biodegradation studies are therefore relevant either to evaluate possibilities of natural attenuation or define bio-remediation strategies. In this study, the possible relationship between the environmental microflora structures and their capabilities for diesel oil biodegradation was investigated. The degradation capacities, i.e. kinetics and extent of biodegradation, were evaluated in closed batch systems by hydrocarbon consumption and CO{sub 2} production, both determined by gas chromatography. The intrinsic biodegradability of different types of diesel oils and the degradation capacities of microflora from ten polluted and ten unpolluted soils samples were determined. The data showed that: i) diesel oil was biodegradable, ii) n-alkanes were totally degraded by each microflora, the final amount of residual hydrocarbons being variable, iii) polluted-soil samples exhibited a slightly higher degradation rate (80%) that polluted-soil samples (67%) or activated sludge (64%). In order to define the contribution of various bacterial groups to diesel oil degradation, enrichment cultures were performed on hydrocarbons representative from the structural classes of diesel oil: hexadecane for n-alkanes, pristane for iso-alkanes, decalin for cyclo-alkanes, phenanthrene for aromatics. By using a 16S rDNA-sequencing method, the bacterial structures of the adapted microflora were determined and compared to that of the native microflora. A marked effect of the selection pressure was observed on the diversity of the microflora, each microflora harboring a major and specific bacterial group. The degradation capacities of the adapted microflora and the occurrence of genes coding for initial hydrocarbon oxidation (alkB, nahAc, cypP450) were also studied. No clear relationship between microflora genes and degradation performances was noted. This seemed to indicate that

  13. Comparison of α-amylase isoforms from the midgut of Cerambyx cerdo L. (Coleoptera: Cerambycidae larvae developed in the wild and on an artificial diet

    Directory of Open Access Journals (Sweden)

    Dojnov Biljana

    2010-01-01

    Full Text Available α-Amylase isoforms of Cerambyx cerdo larvae from the wild (ML and SL and reared in the laboratory (ADL were compared. Three amylase isoforms were presented in the SL and ML extracts while two isoforms were presented in the ADL according to zymogram after isoelectric focusing (IEF. All C. cerdo amylase isoforms were acidic proteins (pI < 3.5. Seven amylase isoforms (ACC 1-7 from the midgut of C. cerdo larvae were found in the ML midgut extract, six in the SL extract, and four in the ADL extract according to native PAGE zymogram. The ADL amylase had the highest activity. All crude midgut extracts of C. cerdo larvae were fractionated on a Superose 12 HR column. The molecular mass of the ACC was estimated to be 34 kDa. .

  14. FEATURES OF THE LARGE INTESTINE MICROFLORA OF CHILDREN – DONOR LIVER TRANSPLANT RECIPIENTS

    Directory of Open Access Journals (Sweden)

    N. I. Gabrielyan

    2013-01-01

    Full Text Available Aim. The study microecology of the large intestine of children with cirrhosis before transplantation of the share liver. Materials and methods. Studied the flora of the colon 157 children of 1 to 17 years admitted to hospital for liver transplantation fragment from a related donor. Identification was carried out using microbial panels BD Crystal and databases BBL Crystal MIND. Methicillin-resistant staphylococci were determined by their sensiti- vity to oxacillin and cefoxitin. Beta-lactamase activity was tested using discs with ceftazidime and ceftazidime/ clavulanic acid. Results. Microecological revealed deep irregularities in the large intestine transplantation in children up lobe of the liver on a spectrum and composition of the microflora. Among the resident microflora decreased levels of bifidobacteria, lactobacilli and coliform bacteria, especially in children under one year. A sig- nificant portion of the children surveyed (over 60–70% had an increase of frequency of finding stateally bacteria, especially Klebsiella and enterobacteria in third children – non-fermenting bacteria – Pseudomonas and Acine- tobacter spp. Revealed the spread of strains of gram-negative bacteria with extended-spectrum betalaktamaz.Conclusion. Expressed microecological violations in the large intestine in children with higher levels of bac- teria are conditionally risk factor reeks of infectious complications in the postoperative period and require are complex tools to assist in eliminatsii.s given antibiotic resistance of bacteria. 

  15. In vitro antimicrobial effects of grape seed extract on peri-implantitis microflora in craniofacial implants

    Institute of Scientific and Technical Information of China (English)

    Binit Shrestha; ML Srithavaj Theerathavaj; Sroisiri Thaweboon; Boonyanit Thaweboon

    2012-01-01

    To determine the antimicrobial effects of grape seed on peri-implantitis microflora.Methods:The grape seed extract was tested against peri-implantitis microflora most commonly found in craniofacial implants including reference strains of Staphylococcus aureus (S. aureus), Escherichia coli (E. coli), Candida albicans (C. albicans) and clinical strains of S. aureus, Klebsiella pneumonia (K. pneumonia) and Candida parapsilosis (C. parapsilosis) by disk diffusion test. Minimum inhibitory concentrations (MIC) and minimum cidal concentrations (MCC) were determined using modified agar dilution millpore method. The extract was further combined with polyethylene glycol and propylene glycol, and was tested for antimicrobial effects. Results: Grape seed extract showed positive inhibitory effects with S. aureus at MIC of 0.625 mg/mL and MCC of 1.25 mg/mL respectively. However the extracts showed minimal or no reactivity against strains of E. coli, K. pneumonia, C. parapsilosis and C. albicans. The use of grape seed extract in combination with polyethylene glycol and propylene glycol also showed dose dependent inhibitory effect on S.aureus. Conclusions: The results of the study showed that grape seed has potential antimicrobial effects which can be further studied and developed to be used in the treatment of infected skin-abutment interface of craniofacial implants.

  16. Effect of dried purslane on immunity responses and cecal microflora composition of broiler chickens

    Directory of Open Access Journals (Sweden)

    Mohammad Reza Ghorbani

    2014-10-01

    Full Text Available The aim of the present study was to evaluate the effect of purslane on immune responses and cecal microflora composition of broiler chickens. One hundred forty four day old broiler chicks (Ross 308 were allocated randomly in 3 groups and 4 replicates to receive diets supplemented with 0 (control, 1 and 2 % of dried purslane. At 21 and 35 days of age 1 mL of sheep red blood cell (SRBC injected to 2 broilers of each replicate and blood samples were taken 7 days after each injection. Eight chicks per treatment at the age of 42 d were killed and the number of coliforms, E-Coli and Lactobacillus bacteria populations in ceca were enumerated on appropriate bacteriological media. There were no significant different in primary and secondary antibody titer against SRBC and no different between the treatments for proportional weight of bursa, thymus and spleen. The results of experiment indicated that use of purslane significantly increased Lactobacillus and decreased E-Coli bacteria population. Use of purslane had no effect on coliforms bacteria population. It is concluded that purslane inclusion had a positive significant effect on cecal microflora composition.

  17. Effects of immune stress on growth performance, immunity, and cecal microflora in chickens.

    Science.gov (United States)

    Yang, X J; Li, W L; Feng, Y; Yao, J H

    2011-12-01

    Immune stress is the loss of immune homeostasis by external forces. This study investigated the effects of different types of immune stress on growth performance, immunity, and the distribution of cecal microflora in broiler chickens. In total, 540 one-day-old Cobb 500 broilers were randomly assigned to receive 1 of 5 (n = 108 birds/group) treatments: 1) no vaccination; 2) simplified vaccination, which included the infectious bronchitis vaccine (H120), the inactivated avian influenza vaccine (AI), the live vaccine strain Clone-30 of the Newcastle disease virus (NDV), and the combined inactive vaccine for infectious bursal diseases and the Newcastle disease vaccine (ND-IB); 3) normal vaccination (simplified vaccination + second dose of ND-IB, H120, and AI); 4) lipopolysaccharide (LPS) stress (normal vaccination+LPS); or 5) cyclophosphamide (CPM) stress (normal vaccination+CPM).The results showed that the average BW and average feed intake decreased significantly after treatment with LPS or CPM (P Chickens that were challenged by LPS or CPM had a lower ileal CP digestibility than that of the control group (P chickens that were treated with LPS or CPM (P chickens were higher than those in the control group chickens at 21 and 42 d of age, respectively (P stress on the microbial populations of treated birds. These data suggest that broilers with simplified vaccinations or without vaccinations can achieve the same growth performance as broilers with general vaccinations, but immune stress can break the homeostasis of cecal microflora and impair intestinal mucosal immune function.

  18. Oatmeal porridge: impact on microflora-associated characteristics in healthy subjects.

    Science.gov (United States)

    Valeur, Jørgen; Puaschitz, Nathalie G; Midtvedt, Tore; Berstad, Arnold

    2016-01-14

    Oatmeal porridge has been consumed for centuries and has several health benefits. We aimed to investigate the effect of oatmeal porridge on gut microflora functions. A total of ten healthy subjects ingested 60 g oatmeal porridge daily for 1 week. The following microflora-associated characteristics were assessed before and after the intervention: intestinal gas production following lactulose ingestion, faecal excretion of SCFA and faecal levels of urease and β-galactosidase. In addition, rectal levels of PGE2 were measured. Microbial fermentation as evaluated by intestinal gas production and excretion of SCFA did not change significantly following the dietary intervention. However, faecal levels of β-galactosidase and urease decreased after eating oatmeal porridge (P=0·049 and 0·031, respectively). Host inflammatory state, as measured by rectal levels of PGE2, also decreased, but the change was not significant (P=0·168). The results suggest that oatmeal porridge has an effect on gut microbial functions and may possess potential prebiotic properties that deserve to be investigated further.

  19. The Cutaneous Bacterial Microflora of the Bodybuilders Using Anabolic-Androgenic Steroids

    Science.gov (United States)

    Zomorodian, Kamiar; Rahimi, Mohammad Javad; Taheri, Mohammad; Ghanbari Asad, Ali; Khani, Soghra; Ahrari, Iman; Pakshir, Keyvan; Khashei, Reza

    2014-01-01

    Background: Anabolic-androgenic steroids (AAS) abuse by the athletes has dramatically increased during the recent decades. These substances might increase the skin lipids and enhance the cutaneous microbial proliferation. Objectives: The current study aimed to investigate the potential side effects of AAS on the bacterial microflora colonization of the bodybuilders` skin. Patients and Methods: The skin samples of 94 male bodybuilders (71 AAS users, 23 non-AAS users) and 46 subjects of the control group, with similar gender and age, were cultured and incubated in both aerobic condition to isolate Staphylococcus aureus and anaerobic condition for Propionibacterium acnes. The isolated bacteria were identified by standard microbiological techniques. Results: The skin lesions were more frequent in the body builders than the controls. Moreover, statistically significant differences were also observed in skin lesions among the AAS users and the non-AAS user athletes. The prevalence of S. aureus and P. acnes in the athletes was higher than that of the control group. In addition, there was a significant difference in distribution of P. acnes between the bodybuilders who used AAS and those who did not. Conclusions: A higher number of bacterial flora was found in the bodybuilders particularly those using AAS in comparison to the controls, which might be due to the influence of these AAS on the skin microflora and transmission of the bacteria through the direct contact of the naked skin with the exercise instruments. PMID:25789120

  20. Autophagy Protects against Colitis by the Maintenance of Normal Gut Microflora and Secretion of Mucus.

    Science.gov (United States)

    Tsuboi, Koichiro; Nishitani, Mayo; Takakura, Atsushi; Imai, Yasuyuki; Komatsu, Masaaki; Kawashima, Hiroto

    2015-08-14

    Genome-wide association studies of inflammatory bowel diseases identified susceptible loci containing an autophagy-related gene. However, the role of autophagy in the colon, a major affected area in inflammatory bowel diseases, is not clear. Here, we show that colonic epithelial cell-specific autophagy-related gene 7 (Atg7) conditional knock-out (cKO) mice showed exacerbation of experimental colitis with more abundant bacterial invasion into the colonic epithelium. Quantitative PCR analysis revealed that cKO mice had abnormal microflora with an increase of some genera. Consistently, expression of antimicrobial or antiparasitic peptides such as angiogenin-4, Relmβ, intelectin-1, and intelectin-2 as well as that of their inducer cytokines was significantly reduced in the cKO mice. Furthermore, secretion of colonic mucins that function as a mucosal barrier against bacterial invasion was also significantly diminished in cKO mice. Taken together, our results indicate that autophagy in colonic epithelial cells protects against colitis by the maintenance of normal gut microflora and secretion of mucus.

  1. Ecological role of algobacterial cenosis links (chlorella - associated microflora or associated bacteria)

    Science.gov (United States)

    Pechurkin, N. S.

    The problems of interrelation of microalgae and bacteria in the "autotroph - heterotroph" aquatic biotic cycle are discussed. The cause and mechanisms of algobacterial cenosis formation still have been explained contradictorily. This work views the results of experimental and theoretical study of algobacterial cenosis functioning by the example of microalga Chlorella vulgaris and associated microflora. The representatives of Pseudomonas mainly predominate in the Chlorella microbial complex. The experiment at non-sterile batch cultivation of Chlorella on Tamya medium showed that the biomass of microorganisms increases simultaneously with the increase of microalgal biomass. Microflora of Chlorella can use organic materials evolved by Chlorella after photosynthesis for reproduction. Moreover, microorganisms can use dying cells of Chlorella, i.e. form the "producer - reducer" biocycle. To understand the cenosis-forming role of microalgae the mathematical model of the "autotroph - heterotroph" aquatic biotic cycle was constructed taking into consideration the opportunities for microorganisms of using Chlorella photosynthates, dying cells and contribution of links to the nitrogen cycle. The theoretical investigation showed that the biomass of associated bacteria growing on glucose and detritus exceeds the biomass of bacteria using only microalgal photosynthates, which is comparable with experimental data.

  2. Isolation of bacterial strains from bovine fecal microflora capable of degradation of ceftiofur.

    Science.gov (United States)

    Rafii, Fatemeh; Williams, Anna J; Park, Miseon; Sims, Lillie M; Heinze, Thomas M; Cerniglia, Carl E; Sutherland, John B

    2009-10-20

    Ceftiofur, a third-generation cephalosporin used to treat bacterial infections in animals, is degraded in bovine feces but the specific bacteria involved are unknown. To find the bacteria involved in ceftiofur metabolism, the bovine fecal microflora was screened. Twenty-one nonidentical strains of bovine fecal bacteria were isolated on media containing 1-32 microg ml(-1) of ceftiofur. The cultures were incubated with 5 microg ml(-1) ceftiofur for different times, then centrifuged and analyzed by high-performance liquid chromatography. Three strains of Bacillus spp., two strains of Roseomonas spp., and one strain of Azospirillum sp. metabolized 5 microg ml(-1) ceftiofur in broth cultures in less than 24h; ten other strains of Roseomonas and one strain of Bacillus pumilus had metabolized it by 120 h. After the ceftiofur had been metabolized by these bacteria, the filter-sterilized supernatants of centrifuged cultures no longer inhibited the growth of a ceftiofur-sensitive strain of Kocuria rhizophila, which indicated that ceftiofur had been transformed to compounds without bactericidal activity. Each isolate was also found to be able to grow in the presence of other beta-lactams, and a nitrocefin assay showed beta-lactamase activity in the 17 strains that metabolized ceftiofur. The results show that some beta-lactamase-producing bacteria from the bovine fecal microflora are capable of transforming ceftiofur to metabolites lacking bactericidal activity.

  3. Prebiotic inulin: Useful dietary adjuncts to manipulate the livestock gut microflora

    Directory of Open Access Journals (Sweden)

    A.K. Samanta

    2013-01-01

    Full Text Available In recent years, there has been a growing appreciation on the relevance of gastrointestinal microflora in both ruminants and non-ruminants owing to revelation of their role in several physiological functions including digestion, nutrient utilization, pathogen exclusion, gastrointestinal development, immunity system, gut gene expression and quality of animal products. The ban imposed on the use of antibiotics and hormones in feed has compelled animal researchers in finding an alternative which could overcome the issues of conventional feed additives. Though the concept of prebiotic was evolved keeping in mind the gastrointestinal flora of human beings, presently animal researchers are exploring the efficiency of prebiotic (inulin for modulating the gut ecosystem of both ruminants and non-ruminants. It was revealed that prebiotic inulin is found to exhibit desirable changes in the gut of non-ruminants like poultry, swine, rabbit etc for augmenting gut health and improvement of product quality. Similarly, in ruminants the prebiotic reduces rumen ammonia nitrogen, methane production, increase microbial protein synthesis and live weight gains in calves. Unlike other feed additives, prebiotic exhibits its effect in multipronged ways for overall increase in the performances of the animals. In coming days, it is expected that prebiotics could be the part of diets in both ruminants and non-ruminants for enabling modulation of gut microflora vis a vis animals productivity in ecological ways.

  4. Prebiotic inulin: Useful dietary adjuncts to manipulate the livestock gut microflora.

    Science.gov (United States)

    Samanta, A K; Jayapal, Natasha; Senani, S; Kolte, A P; Sridhar, Manpal

    2013-05-07

    In recent years, there has been a growing appreciation on the relevance of gastrointestinal microflora in both ruminants and non-ruminants owing to revelation of their role in several physiological functions including digestion, nutrient utilization, pathogen exclusion, gastrointestinal development, immunity system, gut gene expression and quality of animal products. The ban imposed on the use of antibiotics and hormones in feed has compelled animal researchers in finding an alternative which could overcome the issues of conventional feed additives. Though the concept of prebiotic was evolved keeping in mind the gastrointestinal flora of human beings, presently animal researchers are exploring the efficiency of prebiotic (inulin) for modulating the gut ecosystem of both ruminants and non-ruminants. It was revealed that prebiotic inulin is found to exhibit desirable changes in the gut of non-ruminants like poultry, swine, rabbit etc for augmenting gut health and improvement of product quality. Similarly, in ruminants the prebiotic reduces rumen ammonia nitrogen, methane production, increase microbial protein synthesis and live weight gains in calves. Unlike other feed additives, prebiotic exhibits its effect in multipronged ways for overall increase in the performances of the animals. In coming days, it is expected that prebiotics could be the part of diets in both ruminants and non-ruminants for enabling modulation of gut microflora vis a vis animals productivity in ecological ways.

  5. In vitro antimicrobial effects of grape seed extract on peri-implantitis microflora in craniofacial implants

    Institute of Scientific and Technical Information of China (English)

    Binit; Shrestha; M.L.Srithavaj; Theerathavaj; Sroisiri; Thaweboon; Boonyanit; Thaweboon

    2012-01-01

    Objective:To determine the antimicrobial effects of grape seed on peri-implantitis microflora.Methods:The grape seed extract was tested against peri-implantitis microflora most commonly found in craniofacial implants including reference strains of Staphylococcus aureus(S.aureus),Escherichia coli(E.coli),Candida albicans(C.albicans)and clinical strains of S.aureus,Klebsiella pneumonia(K.pneumonia)and Candida parapsilosis(C.parapsilosis)by disk diffusion test.Minimum inhibitory concentrations(MIC)and minimum cidal concentrations(MCC)were determined using modified agar dilution millpore method.The extract was further combined with polyethylene glycol and propylene glycol,and was tested for antimicrobial effects.Results:Grape seed extract showed positive inhibitory effects with S.aureus at MIC of 0.625 mg/mL and MCC of1.25 mg/mL respectively.However the extracts showed minimal or no reactivity against strains of E.coli,K.pneumonia,C.parapsilosis and C.albicans.The use of grape seed extract in combination with polyethylene glycol and propylene glycol also showed dose dependent inhibitory effect on S.aureus.Conclusions:The results of the study showed that grape seed has potential antimicrobial effects which can be further studied and developed to be used in the treatment of infected skinabutment interface of craniofacial implants.

  6. A study of relations between physicochemical properties of crude oils and microbiological characteristics of reservoir microflora

    Science.gov (United States)

    Yashchenko, I. G.; Polishchuk, Yu. M.; Peremitina, T. O.

    2015-10-01

    The dependence of the population and activity of reservoir microflora upon the chemical composition and viscosity of crude oils has been investigated, since it allows the problem of improvement in the technologies and enhancement of oil recovery as applied to production of difficult types of oils with anomalous properties (viscous, heavy, waxy, high resin) to be solved. The effect of the chemical composition of the oil on the number, distribution, and activity of reservoir microflora has been studied using data on the microbiological properties of reservoir water of 16 different fields in oil and gas basins of Russia, Mongolia, China, and Vietnam. Information on the physicochemical properties of crude oils of these fields has been obtained from the database created at the Institute of Petroleum Chemistry, Siberian Branch on the physicochemical properties of oils throughout the world. It has been found that formation water in viscous oil reservoirs is char acterized by a large population of heterotrophic and sulfate reducing bacteria and the water of oil fields with a high paraffin content, by population of denitrifying bacteria.

  7. Metabolism of stevioside and rebaudioside A from Stevia rebaudiana extracts by human microflora.

    Science.gov (United States)

    Gardana, Claudio; Simonetti, Paolo; Canzi, Enrica; Zanchi, Raffaella; Pietta, Piergiorgio

    2003-10-22

    Stevia rebaudiana standardized extracts (SSEs) are used as natural sweeteners or dietary supplements in different countries for their content of stevioside or rebaudioside A. These compounds possess up to 250 times the sweetness intensity of sucrose, and they are noncaloric and noncariogenic sweeteners. The aim of this study was to investigate the in vitro transformation of stevioside and rebaudioside A after incubation with human microflora, the influence of these sweeteners on human microbial fecal community and which specific groups metabolize preferentially stevioside and rebaudioside A. The experiments were carried out under strict anaerobic conditions in batch cultures inoculated with mixed fecal bacteria from volunteers. The hydrolysis was monitored by HPLC coupled to photodiode array and mass spectrometric detectors. Isolated bacterial strains from fecal materials incubated in selective broths were added to stevioside and rebaudioside A. These sweeteners were completely hydrolyzed to their aglycon steviol in 10 and 24 h, respectively. Interestingly, the human intestinal microflora was not able to degrade steviol. Furthermore, stevioside and rebaudioside A did not significantly influence the composition of fecal cultures; among the selected intestinal groups, bacteroides were the most efficient in hydrolyzing Stevia sweeteners to steviol.

  8. 肠道菌群与代谢性疾病%Intestinal microflora and metabolic diseases

    Institute of Scientific and Technical Information of China (English)

    解傲; 袁杰利

    2013-01-01

    Intestinal microflora has a large amount and are of various species, which participate in most metabolic activities in vivo performing considerable effects. It has been drawing many attentions to obesity, hypercholesteremia, hypertension, diabetes caused by high-fat and high-protein diet in these days. Results from the majority of studies suggested there is a connection between microflora and metabolic diseases, and this paper reviews it.%肠道微生物数量庞大、种类繁多,在机体中参与了各种代谢活动,发挥着重要的作用.近年来,随着人们生活水平的提高,饮食结构偏重于高脂肪、高蛋白,导致了肥胖以及"三高"等代谢性疾病,引起了科研工作者得广泛关注.大部分研究结果显示,肠道菌群与代谢性疾病存在着一定的联系,本文针对此问题作一综述.

  9. First insight into the lipid uptake, storage and mobilization in arachnids: role of midgut diverticula and lipoproteins.

    Science.gov (United States)

    Laino, Aldana; Cunningham, Mónica L; García, Fernando; Heras, Horacio

    2009-12-01

    The importance of midgut diverticula (M-diverticula) and hemolymph lipoproteins in the lipid homeostasis of Polybetes phythagoricus was studied. Radioactivity distribution in tissues and hemolymph was analyzed either after feeding or injecting [1-(14)C]-palmitate. In both experiments, radioactivity was mostly taken up by M-diverticula that synthesized diacylglycerols, triacylglycerols and phospholipids in a ratio close to its lipid class composition. M-diverticula total lipids represent 8.08% (by wt), mostly triacylglycerols (74%) and phosphatidylcholine (13%). Major fatty acids were (in decreasing order of abundance) 18:1n-9, 18:2n-6, 16:0, 16:1n-7, 18:0, 18:3n-3. Spider hemocyanin-containing lipoprotein (VHDL) transported 83% of the circulating label at short incubation times. After 24h, VHDL and HDL-1 (comparable to insect lipophorin) were found to be involved in the lipid uptake and release from M-diverticula, HDL-2 playing a negligible role. Lipoprotein's labelled lipid changed with time, phospholipids becoming the main circulating lipid after 24h. These results indicate that arachnid M-diverticula play a central role in lipid synthesis, storage and movilization, analogous to insect fat body or crustacean midgut gland. The relative contribution of HDL-1 and VHDL to lipid dynamics indicated that, unlike insects, spider VHDL significantly contributes to the lipid exchange between M-diverticula and hemolymph.

  10. Histopathological effects and determination of the putative receptor of Bacillus thuringiensis Cry1Da toxin in Spodoptera littoralis midgut.

    Science.gov (United States)

    BenFarhat-Touzri, Dalel; Saadaoui, Marwa; Abdelkefi-Mesrati, Lobna; Saadaoui, Imen; Azzouz, Hichem; Tounsi, Slim

    2013-02-01

    Bacillus thuringiensis subsp. aizawai strain HD133, known by its effectiveness against Spodoptera species, produces many insecticidal proteins including Cry1Ab, Cry1Ca and Cry1Da. In the present study, the insecticidal activity of Cry1Da against Spodoptera littoralis was investigated. It showed toxicity with an LC(50) of 224.4 ng/cm(2) with 95% confidence limits of (178.61-270.19) and an LC(90) of 467.77 ng/cm(2) with 95% confidence limits of (392.89-542.65). The midgut histopathology of Cry1Da fed larvae showed vesicle formation in the apical region, vacuolization and destruction of epithelial cells. Biotinylated-activated Cry1Da toxin bound protein of about 65 kDa on blots of S. littoralis brush border membrane preparations. This putative receptor differs in molecular size from those recognized by Cry1C and Vip3A which are active against this polyphagous insect. This difference in midgut receptors strongly supports the use of Cry1Da as insecticidal agent, particularly in case of Cry and/or Vip-resistance management.

  11. Exploring the midgut transcriptome and brush border membrane vesicle proteome of the rice stem borer, Chilo suppressalis (Walker.

    Directory of Open Access Journals (Sweden)

    Weihua Ma

    Full Text Available The rice stem borer, Chilo suppressalis (Walker (Lepidoptera: Pyralidae, is one of the most detrimental pests affecting rice crops. The use of Bacillus thuringiensis (Bt toxins has been explored as a means to control this pest, but the potential for C. suppressalis to develop resistance to Bt toxins makes this approach problematic. Few C. suppressalis gene sequences are known, which makes in-depth study of gene function difficult. Herein, we sequenced the midgut transcriptome of the rice stem borer. In total, 37,040 contigs were obtained, with a mean size of 497 bp. As expected, the transcripts of C. suppressalis shared high similarity with arthropod genes. Gene ontology and KEGG analysis were used to classify the gene functions in C. suppressalis. Using the midgut transcriptome data, we conducted a proteome analysis to identify proteins expressed abundantly in the brush border membrane vesicles (BBMV. Of the 100 top abundant proteins that were excised and subjected to mass spectrometry analysis, 74 share high similarity with known proteins. Among these proteins, Western blot analysis showed that Aminopeptidase N and EH domain-containing protein have the binding activities with Bt-toxin Cry1Ac. These data provide invaluable information about the gene sequences of C. suppressalis and the proteins that bind with Cry1Ac.

  12. Lipase and invertase activities in midgut and salivary glands of Chilo suppressalis (Walker (Lepidoptera, Pyralidae, rice striped stem borer

    Directory of Open Access Journals (Sweden)

    A Zibaee

    2008-11-01

    Full Text Available The rice striped stem borer, Chilo supprressalis, was introduced to Iran in 1973 where it is now widely distributed and causes severe damages. Lipases, which catalyses the hydrolysis of fatty acid ester bonds, are widely distributed among animals, plants and microorganisms. Invertases (β-fructofuranosidase are glycosidehydrolases that catalyze the cleavage of sucrose (β-D-glucopyranosyl-S-D-fructofuranoside into the monosaccharides glucose and fructose. Laboratory-reared 4th instar larvae were randomly selected, their midgut and salivary glands were removed by dissection under a light microscope and lipase and invertase activities were assayed. The activity of lipase/invertase in the midgut and salivary gland were 0.49/0.27 and 0.35/0.23 μmol/min/mg protein, respectively. The optimum pH and temperature for both the two enzymes were determined to be 10-11 and 37-40 °C, which is consistent with pH and temperature values already observed in Lepidoptera. The enzyme activity was reduced by addition of NaCl, KCl, MgCl2, SDS, urea and plant extracts from Artemisia annua, but not by CaCl2 which enhanced enzyme activity. Pest control with usage of resistant varieties of plants is one of the most important practices that are dependant on inhibitors already present in nature. Hence, characterization of insect digestive enzymes, especially examination of inhibition effects on enzyme activity, could be useful in developing new strategies for pest control.

  13. Exploring the midgut transcriptome and brush border membrane vesicle proteome of the rice stem borer, Chilo suppressalis (Walker).

    Science.gov (United States)

    Ma, Weihua; Zhang, Zan; Peng, Chuanhua; Wang, Xiaoping; Li, Fei; Lin, Yongjun

    2012-01-01

    The rice stem borer, Chilo suppressalis (Walker) (Lepidoptera: Pyralidae), is one of the most detrimental pests affecting rice crops. The use of Bacillus thuringiensis (Bt) toxins has been explored as a means to control this pest, but the potential for C. suppressalis to develop resistance to Bt toxins makes this approach problematic. Few C. suppressalis gene sequences are known, which makes in-depth study of gene function difficult. Herein, we sequenced the midgut transcriptome of the rice stem borer. In total, 37,040 contigs were obtained, with a mean size of 497 bp. As expected, the transcripts of C. suppressalis shared high similarity with arthropod genes. Gene ontology and KEGG analysis were used to classify the gene functions in C. suppressalis. Using the midgut transcriptome data, we conducted a proteome analysis to identify proteins expressed abundantly in the brush border membrane vesicles (BBMV). Of the 100 top abundant proteins that were excised and subjected to mass spectrometry analysis, 74 share high similarity with known proteins. Among these proteins, Western blot analysis showed that Aminopeptidase N and EH domain-containing protein have the binding activities with Bt-toxin Cry1Ac. These data provide invaluable information about the gene sequences of C. suppressalis and the proteins that bind with Cry1Ac.

  14. Feasibility study on the application of rhizosphere microflora of rice for the biohydrogen production from wasted bread

    Energy Technology Data Exchange (ETDEWEB)

    Doi, Tetsuya [Field Production Science Center, Graduate School of Agriculture and Life Sciences, The University of Tokyo, Nishitokyo, Tokyo 188-0002 (Japan); Nishihara Environment Technology Inc., Tokyo 108-0023 (Japan); Matsumoto, Hisami [Nishihara Environment Technology Inc., Tokyo 108-0023 (Japan); Abe, Jun [AE-Bio, Graduate School of Agriculture and Life Sciences, The University of Tokyo, Tokyo 113-8657 (Japan); Morita, Shigenori [Field Production Science Center, Graduate School of Agriculture and Life Sciences, The University of Tokyo, Nishitokyo, Tokyo 188-0002 (Japan)

    2009-02-15

    We performed an experiment of continuous anaerobic hydrogen fermentation as a pilot-plant-scale test, in which waste from a bread factory was fermented by microflora of rice rhizosphere origin. The community structure of microflora during anaerobic hydrogen fermentation was analyzed using PCR-DGGE, FISH, and quinone profiles. The relation of those results to hydrogen generation was discussed. Results show that a suitable condition was a reactor temperature of 35 C, with HRT 12-36 h, volume load of 30-70 kg-COD{sub Cr}/m{sup 3} day, and maximum hydrogen production rate of 1.30 mol-H{sub 2}/mol-hexose. Regarding characteristics of microflora during fermentation, PCR-DGGE results show specific 16S rDNA band patterns; Megasphaera elsdenii and Clostridium sp. of the hydrogen-producing bacteria were identified. M. elsdenii was detected throughout the fermentation period, while Clostridium sp. of hydrogen-producing bacteria was detected on the 46th day. Furthermore, FISH revealed large amounts of Clostridium spp. in the sample. The quinone profile showed that the dominant molecular species of quinone is MK-7. Because Clostridium spp. belong to MK-7, results suggest that the quinone profile result agrees with the results of PCR-DGGE and FISH. Microflora in the rhizosphere of rice plants can be a possible resource for effective bacteria of biohydrogen production. (author)

  15. Effect of fermentation temperature on hydrogen production from cow waste slurry by using anaerobic microflora within the slurry.

    Science.gov (United States)

    Yokoyama, Hiroshi; Waki, Miyoko; Moriya, Naoko; Yasuda, Tomoko; Tanaka, Yasuo; Haga, Kiyonori

    2007-02-01

    We examined hydrogen production from a dairy cow waste slurry (13.4 g of volatile solids per liter) by batch cultures in a temperature range from 37 to 85 degrees C, using microflora naturally present within the slurry. Without the addition of seed bacteria, hydrogen was produced by simply incubating the slurry, using the microflora within the slurry. Interestingly, two peaks of fermentation temperatures for hydrogen production from the slurry were observed at 60 and 75 degrees C (392 and 248 ml H2 per liter of slurry, respectively). After the termination of the hydrogen evolution, the microflora cultured at 60 degrees C displayed hydrogen-consuming activity, but hydrogen-consuming activity of the microflora cultured at 75 degrees C was not detected, at least for 24 days. At both 60 and 75 degrees C, the main by-product was acetate, and the optimum pH of the slurry for hydrogen production was around neutral. Bacteria related to hydrogen-producing moderate and extreme thermophiles, Clostridium thermocellum and Caldanaerobacter subterraneus, were detected in the slurries cultured at 60 and 75 degrees C, respectively, by denaturing gradient gel electrophoresis analyses, using the V3 region of 16S rDNA.

  16. Microbial DNA fingerprinting of human fingerprints: dynamic colonization of fingertip microflora challenges human host inferences for forensic purposes

    NARCIS (Netherlands)

    Tims, S.; Wamel, van W.; Endtz, H.P.; Belkum, van A.; Kayser, M.

    2010-01-01

    Human fingertip microflora is transferred to touched objects and may provide forensically relevant information on individual hosts, such as on geographic origins, if endogenous microbial skin species/strains would be retrievable from physical fingerprints and would carry geographically restricted DN

  17. Microbial DNA fingerprinting of human fingerprints: dynamic colonization of fingertip microflora challenges human host inferences for forensic purposes

    NARCIS (Netherlands)

    S. Tims (Sebastian); W.J.B. van Wamel (Willem); H.P. Endtz (Hubert); A.F. van Belkum (Alex); M.H. Kayser (Manfred)

    2009-01-01

    textabstractHuman fingertip microflora is transferred to touched objects and may provide forensically relevant information on individual hosts, such as on geographic origins, if endogenous microbial skin species/strains would be retrievable from physical fingerprints and would carry geographically r

  18. Microbial DNA fingerprinting of human fingerprints : dynamic colonization of fingertip microflora challenges human host inferences for forensic purposes

    NARCIS (Netherlands)

    Tims, Sebastian; van Wamel, Willem; Endtz, Hubert P.; van Belkum, Alex; Kayser, Manfred

    2010-01-01

    Human fingertip microflora is transferred to touched objects and may provide forensically relevant information on individual hosts, such as on geographic origins, if endogenous microbial skin species/strains would be retrievable from physical fingerprints and would carry geographically restricted DN

  19. [Indigenous microflora associated with the tegument of Triaenophorus nodulosus (Cestoda) and the intestine of its pike host].

    Science.gov (United States)

    Korneva, Zh V; Plotnikov, A O

    2006-01-01

    Investigations of the indigenous microflora associated with the mucous intestines of fish and its cestode parasites have been for the first time carried out using the methods of transmission and scanning electron microscopy. New data on the bacterial biodiversity in the cestode and its fish host are obtained. Nanobacteria and spirochaetes are for the first time revealed in a fish host together with the previously known bacteria forming the intestinal microflora of fish. Spirochaetes were shown to be associated with the intestines of a pike host only, while nanobacteria cover abundantly the surface of the apical parts of the intestinal microvilli and the apical parts of the microtriches in the cestode tegument. The similarity of the bacterial floras associated with the apical surface of the parasite tegument and the intestine of the host should be noted. At the same time, deeper bacterial communities represented by obligate symbionts are specific. Thus, there is a normal indigenous microflora in cestodes, associated with the tegumental surface. This symbiotic microflora has specific morphological features and provides the balance of relations in the parasite-host system.

  20. Oral administration of Bifidobacterim bifidum for modulating microflora, acid and bile resistance, and physiological indices in mice.

    Science.gov (United States)

    Wang, Bao-Gui; Xu, Hai-Bo; Wei, Hua; Zeng, Zhe-Ling; Xu, Feng

    2015-02-01

    Bifidobacteria are generally acknowledged as major gut microflora used as probiotics, which promote human health. In this study, the effects of the administration of Bifidobacterim bifidum on modulating gastrointestinal (GI) tract microflora, acid and bile resistance, and physiological indices in BALB/c mice were investigated. Results showed that B. bifidum can significantly improve the ecosystem of the GI tract by increasing the amount of probiotics and reducing the populations of pathogenic bacteria, as measured by plate count and real-time PCR. After exposure to simulated GI tract conditions, the growth of gut microflora in the B. bifidum group was higher than that in the control group when incubated for 12 h in MRS or nutrient broth adjusted to pH 2.0 or 3.0 or in the presence of a concentration of bile salt (0.45% m/v). The blood biochemical index was examined, and the physiological effect of the cell-free extract of gut microflora was evaluated by measuring the activity of various enzymes, including α-glucosidases, esterase, and lactate dehydrogenase. This study suggested that a B. bifidum strain can stabilize blood sugar, lower cholesterol levels in serum, and improve metabolic activity. Moreover, B. bifidum was a promising enhancer of microbial diversity in mouse intestine and played a vital role in human physiological processes, which can benefit the health of a host.

  1. MICROBIAL LANDSCAPE OF MICROFLORA OF A PHARYNX AT PATIENTS WITH TONSILLIT’S PATHOLOGY

    Directory of Open Access Journals (Sweden)

    O. Yu. Borisova

    2015-01-01

    Full Text Available The microbial landscape of microflora of a pharynx at patients with tonsillit’s pathology were studied. Materials and methods. 79 patients from GBUZ “Research Institute of Clinical Otorhinolaryngology” (78.5% patients with various forms of chronic tonsillitis and 21.5% of patients without chronic tonsillitis (control group. Among patients of the main group with chronic tonsillitis, at 60 (96.8% patients there was a diagnosis chronic tonsillitis a toxical-allergic form of 1 degree (TAF I, at 1 (1.6% the patient — chronic tonsillitis a toxical-allergic form II of degree (TAF II and at 1 (1.6% the patient — chronic tonsillitis. Identification of microorganisms carried out on cultural-morphological and biochemical properties. Specific identification of the hardly cultivated microorganisms and Corynebacterium was carried out by MALDI-TOFF MS of BioMerieux VITEK MS MALDI-TOF (“bioMerieux”, France. Identification of the allocated Corynebacterium was carried out by amplification of a gene rpoB and the subsequent direct sequencing. Results. The majority (98.7% of the allocated microorganisms, treated 27 types and were Gram-positive. It is revealed 159 strains of 29 species of microorganisms, from them 41.4% of strains belonged to the Streptococcus, 19.7% — Staphylococcus, 36.9% — Corynebacterium. Among Streptococcus — 55.4% of S. viridans, 38.4% — S. pyogenes and 3.1% — of S. pneumoniae и S. oralis; Staphylococcus — 64.5% of S. aureus, 32.2% of S. epidermidis and 3.3% of S. hominis. 18 types of Corynebacterium — С. tuberculostearicum (17.2% strains, C. рseudodiphtheriticum (15.5% strains and C. aurimucosum (18.9% strains are revealed. At 44.3% of the surveyed patients the microflora is presented by a monoculture and at 55.7% associations are revealed. Conclusion. The microflora at patients with tonsillit’s pathology is characterized. At the expressed pathological process in a microbiota of a pharynx the monoculture while

  2. Effects of neem oil (Azadirachta indica A. Juss) on the replacement of the midgut epithelium in the lacewing Ceraeochrysa claveri during larval-pupal metamorphosis.

    Science.gov (United States)

    Scudeler, Elton Luiz; Padovani, Carlos Roberto; Santos, Daniela Carvalho Dos

    2014-06-01

    Larvae of the lacewing Ceraeochrysa claveri were fed on eggs of Diatraeasaccharalis treated with neem oil at concentrations of 0.5%, 1% and 2% throughout the larval period. Pupae obtained from treated larvae were used in the study at five days after the completion of cocoon spinning to investigate the effects of neem oil on the replacement of the midgut epithelium during the larval-pupal transition. We observed that the old larval epithelium was shed into the midgut lumen and transformed into the yellow body. Old cells from the yellow body were destroyed by apoptosis and autophagy and were not affected by neem oil. However, neem oil did affect the new pupal epithelium. Cells from treated pupae showed cellular injuries such as a loss of microvilli, cytoplasmic vacuolization, an increase of glycogen stores, deformation of the rough endoplasmic reticulum and dilation of the perinuclear space. Additionally, the neem oil treatment resulted in the release of cytoplasmic protrusions, rupture of the plasma membrane and leakage of cellular debris into the midgut lumen, characteristics of cell death by necrosis. The results indicate that neem oil ingestion affects the replacement of midgut epithelium, causing cytotoxic effects that can alter the organism's physiology due to extensive cellular injuries.

  3. Cloning eleven midgut trypsin cDNAs and evaluating the interaction of proteinase inhibitors with Cry1Ac against the tobacco budworm Heliothis virescens (F.) (Lepidoptera: Noctuidae)

    Science.gov (United States)

    Midgut trypsins are associated with Bt protoxin activation and toxin degradation. Proteinase inhibitors have potential insecticidal toxicity against a wide range of insect species. Proactive action to examine trypsin gene profiles and proteinase inhibitors for interaction with Bt toxin is necessary ...

  4. Expression of a sugar clade gustatory receptor, BmGr6, in the oral sensory organs, midgut, and central nervous system of larvae of the silkworm Bombyx mori.

    Science.gov (United States)

    Mang, Dingze; Shu, Min; Endo, Haruka; Yoshizawa, Yasutaka; Nagata, Shinji; Kikuta, Shingo; Sato, Ryoichi

    2016-03-01

    Insects taste nonvolatile chemicals through gustatory receptors (Grs) and make choices for feeding, mating, and oviposition. To date, genome projects have identified 69 Gr genes in the silkworm, Bombyx mori; however, the expression sites of these Grs remain to be explored. In this study, we used reverse transcription (RT)-PCR to investigate expression of the B. mori Gr-6 (BmGr6) gene, a member of the putative sugar clade gene family in various tissues. BmGr6 is expressed in the midgut, central nervous system (CNS), and oral sensory organs. Moreover, immunohistochemistry using an anti-BmGr6 antiserum demonstrated that BmGr6 is expressed in cells by oral sensory organs, midgut and nervous system. Furthermore, double-immunohistochemistry indicated that BmGr6 is expressed in midgut enteroendocrine cells, also in CNS neurosecretory cells. In particular, a portion of BmGr6-expressing cells, in both midgut and CNS, secretes FMRFamide-related peptides (FaRPs). These results suggest that BmGr6 functions not only as a taste receptor, but also as a chemical sensor such as for the regulation of gut movement, physiological conditions, and feeding behavior of larvae.

  5. Ultrastructural studies of the termite (Odontotermes obesus) gut microflora and its cellulolytic properties.

    Science.gov (United States)

    Paul, J; Saxena, S; Varma, A

    1993-01-01

    The major gut microflora colonizing the hind gut of a higher termite,Odontotermes obesus, included morphologically diverse bacteria, both coccoid and rod-shaped, along with spirochaetes, pseudomonads and actinomycetes. Flagellated protozoa were totally absent. When the gut extract was inoculated on plates containing carboxymethyl cellulose or cellobiose, higher numbers of bacteria grew than on plates without cellulosic sources. The gut homogenate exhibited strong hydrolytic activity when carboxymethyl cellulose,p-nitrophenyl-β-D-glucoside or xylan were used as substrate, indicating the role of gut microbiota in the process of cellulose and hemicellulose digestion. Activities were highest in the hind gut, and the paunch was probably the major site of polysaccharide digestion in this higher termite.In vitro cultivation of some of the isolates revealed both cellulase and xylanase activities. To our knowledge, this is the first report on ultrastructural studies of the higher termiteOdontotermes obesus.

  6. The Effects of GH Transgenic Goats on the Microflora of the Intestine, Feces and Surrounding Soil.

    Science.gov (United States)

    Bao, Zekun; Gao, Xue; Zhang, Qiang; Lin, Jian; Hu, Weiwei; Yu, Huiqing; Chen, Jianquan; Yang, Qian; Yu, Qinghua

    2015-01-01

    The development of genetically engineered animals has brought with it increasing concerns about biosafety issues. We therefore evaluated the risks of growth hormone from transgenic goats, including the probability of horizontal gene transfer and the impact on the microbial community of the goats' gastrointestinal tracts, feces and the surrounding soil. The results showed that neither the GH nor the neoR gene could be detected in the samples. Moreover, there was no significant change in the microbial community of the gastrointestinal tracts, feces and soil, as tested with PCR-denaturing gradient gel electrophoresis and 16S rDNA sequencing. Finally, phylogenetic analysis showed that the intestinal content, feces and soil samples all contained the same dominant group of bacteria. These results demonstrated that expression of goat growth hormone in the mammary of GH transgenic goat does not influence the microflora of the intestine, feces and surrounding soil.

  7. The Effects of GH Transgenic Goats on the Microflora of the Intestine, Feces and Surrounding Soil.

    Directory of Open Access Journals (Sweden)

    Zekun Bao

    Full Text Available The development of genetically engineered animals has brought with it increasing concerns about biosafety issues. We therefore evaluated the risks of growth hormone from transgenic goats, including the probability of horizontal gene transfer and the impact on the microbial community of the goats' gastrointestinal tracts, feces and the surrounding soil. The results showed that neither the GH nor the neoR gene could be detected in the samples. Moreover, there was no significant change in the microbial community of the gastrointestinal tracts, feces and soil, as tested with PCR-denaturing gradient gel electrophoresis and 16S rDNA sequencing. Finally, phylogenetic analysis showed that the intestinal content, feces and soil samples all contained the same dominant group of bacteria. These results demonstrated that expression of goat growth hormone in the mammary of GH transgenic goat does not influence the microflora of the intestine, feces and surrounding soil.

  8. Citric Acid Production by the Aspergillus niger Isolated from the Microflora of Iran

    Directory of Open Access Journals (Sweden)

    R.Yazdanparast

    1995-08-01

    Full Text Available Citric acid production by A.niger, isolated from the microflora of Iran, has been investigated in liquid and semi-solid states using growth media with different compositions. In 2% media made of Rocheh grape pomace or sabouraud dextrose, the yield of citric acid production was 0.7 g per Kg of the pomace; and the yield decreased by 50% in 2% saghal solian grape pomace medium. However, in 40% (W/W saghal solian semi-solid medium containing 3% methanol, the yield of citric acid production has improved to 80 g per Kg of pomace in stationary mode of production and to 120 g per Kg of pomace in the rolling mode of fermentation.

  9. NORMAL NASOPHARYNGEAL MICROFLORA AS A RESERVOIR OF MULTIRESISTANT STRAINS OF UPPER RESPIRATORY TRACT INFECTIONS

    Directory of Open Access Journals (Sweden)

    Minukhin V.V.

    2015-05-01

    Full Text Available Nasopharinheal carriage of bacteria may play a central role in the development and spread of respiratory infections. In addition, so-called "healthy" carriage is often transformed under the influence of various factors into an active infection.It is necessary to take into account not only the range of possible pathogens, but also trends in the development of antibiotic resistance of leading etiologic agents while choosing tactics of antimicrobial therapy. The investigation was designed to study the role of normal microflora of the nasopharynx as a reservoir of resistant strains of respiratory infections. Materials and Methods. Fifty three healthy individuals and 168 patients with acute upper respiratory tract infections who had been treated in CEHC "Kharkiv Municipal Clinical Hospital № 30" were examined. Microbiological study included isolation and identification of pathogens in accordance with the Order of the Ministry of Health Care № 535 from 22.04.1985., determination of the sensitivity of microorganisms to antibiotics by diffusion method according to the Order of the Ministry of Health Care of Ukraine № 167 from 05.04.2007. Results and discussion. Bacteriological study of nasal swabs of healthy people showed that the composition of the microflora of the nasopharynx contained potentially pathogenic microorganisms. Among the isolated microorganisms essential place was occupied by S. epidermidis and S. aureus, both in monoculture and association. Epidermal staphylococcus was isolated in 36 % and Staphylococcus aureus in 27% of cases. Pneumococcus and hemolytic streptococcus of group A were isolated in 23 and 14% of cases, respectively. One hundred and eighty strains of opportunistic microorganisms were isolated in the study of nasopharyngeal microflora of patients with acute upper respiratory tract infection. The leading role belonged to S. pyogenes (40.5% and S.epidermidis (33,3%. S. aureus (12,8% and S.pneumoniae (10,6% were next

  10. Characterisation of the gut microflora in rainbow trout fry (Oncorhynchus mykiss) using deep-sequencing

    DEFF Research Database (Denmark)

    Ingerslev, Hans-Christian; Dalsgaard, Inger; Boye, Mette;

    2012-01-01

    For many years it has been known that the bacterial microflora in the gut of warm-blooded animals exists in harmony with the host and exert various beneficial effects on the health by their metabolic activities. Hence, the gut microbiota has a high importance for the animal. In many studies from e...... was used to examine the composition of the microbial flora in the gut of rainbow trout (Oncorhynchus mykiss) fry. The fish were examined before and after first-feeding and after administration by commercial probiotic lactobacilli to the feed........g. humans and the pig mapping of the bacterial flora from the gut have shown dominance by some specific bacterial groups, and this bacterial profile is termed as a ‘core microbiota’. For lower vertebrates like fish mapping of the bacterial flora in the gastrointestinal system is to date a relatively new...

  11. Polymeric proanthocyanidins are catabolized by human colonic microflora into low-molecular-weight phenolic acids.

    Science.gov (United States)

    Déprez, S; Brezillon, C; Rabot, S; Philippe, C; Mila, I; Lapierre, C; Scalbert, A

    2000-11-01

    Polymeric proanthocyanidins are common constituents of many foods and beverages. Their fate in the human body remains largely unknown. Their metabolism by human colonic microflora incubated in vitro in anoxic conditions has been investigated using nonlabeled and (14)C-labeled purified proanthocyanidin polymers. Polymers were almost totally degraded after 48 h of incubation. Phenylacetic, phenylpropionic and phenylvaleric acids, monohydroxylated mainly in the meta or para position, were identified as metabolites by gas chromatography coupled to mass spectrometry (GC-MS). Yields were similar to those previously reported for flavonoid monomers. These results provide the first evidence of degradation of dietary phenolic polymers into low-molecular-weight aromatic compounds. To understand the nutritional properties of proanthocyanidins, it is therefore essential to consider the biological properties of these metabolites.

  12. [Unknown pathogens from the human oral microflora of interest for otorhinolaryngology].

    Science.gov (United States)

    Tets, V V; Tets, G V; Vikina, D S; Vecherovskaia, M F; Kharlamova, V V

    2014-01-01

    The identification of microorganisms from the human oral cavity is a topical problem in many clinical disciplines including otorhinolaryngology. Many bacterial species of oral microbiota are causative agents of ENT diseases, and poor conditions of the oral cavity, unhealthy teeth and gums increase the risk of the spread of infection. The objective of the present study was to distinguish and identify poorly explored and previously unknown aerobic opportunistic pathogenic microorganisms responsible for various ENT disorders. The normal microflora remains to be thoroughly studied with the use of the new culturing techniques that ensure the isolation of pure microbial cultures. The present publication reports for the first time isolation of the stable mixed microbial biofilms formed by unrelated bacterial species and containing the yet unknown microorganisms. The isolated bacteria were identified as previously unknown or poorly unexplored anaerobic opportunistic species capable of inducing ENT pathology; their sensitivity to antibiotics was evaluated.

  13. Concomitant Antibiotic and Mercury Resistance Among Gastrointestinal Microflora of Feral Brook Trout, Salvelinus fontinalis

    Science.gov (United States)

    Meredith, Matthew M.; Parry, Erin M.; Guay, Justin A.; Markham, Nicholas O.; Danner, G. Russell; Johnson, Keith A.; Barkay, Tamar; Fekete, Frank A.

    2013-01-01

    Twenty-nine bacterial isolates representing eight genera from the gastrointestinal tracts of feral brook trout Salvelinus fontinalis (Mitchell) demonstrated multiple maximal antibiotic resistances and concomitant broad-spectrum mercury (Hg) resistance. Equivalent viable plate counts on tryptic soy agar supplemented with either 0 or 25 μM HgCl2 verified the ubiquity of mercury resistance in this microbial environment. Mercury levels in lake water samples measured 1.5 ng L−1; mercury concentrations in fish filets ranged from 81.8 to 1,080 ng g−1 and correlated with fish length. The presence of similar antibiotic and Hg resistance patterns in multiple genera of gastrointestinal microflora supports a growing body of research that multiple selective genes can be transferred horizontally in the presence of an unrelated individual selective pressure. We present data that bioaccumulation of non-point source Hg pollution could be a selective pressure to accumulate both antibiotic and Hg resistant bacteria. PMID:22850694

  14. Efficacy of oral Bifidobacterium bifidum ATCC 29521 on microflora and antioxidant in mice.

    Science.gov (United States)

    Wang, Bao-gui; Xu, Hai-bo; Xu, Feng; Zeng, Zhe-ling; Wei, Hua

    2016-03-01

    This study aimed to examine whether Bifidobacterium bifidum ATCC 29521, a species of colonic microflora in humans, is involved in the intestinal tract of mice. This study was also conducted to determine the antioxidant activity of this species by evaluating different microbial populations and reactive oxygen species isolated from feces and intestinal contents for 28 days of oral administration. Microbial diversities were assessed through bacterial culture techniques, PCR-DGGE, and real-time PCR. This study showed that the intake of B. bifidum ATCC 29521 significantly (p stress were also determined. Results indicated that B. bifidum ATCC 29521 elicits a beneficial effect on murine gut microbiota and antioxidant activities compared with the control samples. This species can be considered as a potential bioresource antioxidant to promote health. Bifidobacterium bifidum ATCC 29521 may also be used as a promising material in microbiological and food applications.

  15. 肝移植与肠道微生态%Liver transplantation and intestinal microflora

    Institute of Scientific and Technical Information of China (English)

    李政锋; 阮冰

    2012-01-01

    肠道黏膜与肠道内共生的微生物形成了肠道黏膜屏障,可有效防止内源性及外源性微生物及毒素的侵害;肝脏与肠道关系密切,作为终末期肝病的有效治疗手段,肝移植术打破了原有的平衡,造成了肠道微生态的改变.%Intestinal mucosa and gut microbes form the intestinal mucosal barrier, which effectively prevent the damages resulted from endogenous and exogenous microorganism or toxins. The liver is closely related to intestinal tract. As the effective treatments for end-stage liver disease, liver transplantation broke down the original balance, and changed the intestinal microflora.

  16. Determination of nitric oxide metabolites, nitrate and nitrite, in Anopheles culicifacies mosquito midgut and haemolymph by anion exchange high-performance liquid chromatography: plausible mechanism of refractoriness

    Directory of Open Access Journals (Sweden)

    Adak Tridibesh

    2008-04-01

    Full Text Available Abstract Background The diverse physiological and pathological role of nitric oxide in innate immune defenses against many intra and extracellular pathogens, have led to the development of various methods for determining nitric oxide (NO synthesis. NO metabolites, nitrite (NO2- and nitrate (NO3- are produced by the action of an inducible Anopheles culicifacies NO synthase (AcNOS in mosquito mid-guts and may be central to anti-parasitic arsenal of these mosquitoes. Method While exploring a plausible mechanism of refractoriness based on nitric oxide synthase physiology among the sibling species of An. culicifacies, a sensitive, specific and cost effective high performance liquid chromatography (HPLC method was developed, which is not influenced by the presence of biogenic amines, for the determination of NO2- and NO3- from mosquito mid-guts and haemolymph. Results This method is based on extraction, efficiency, assay reproducibility and contaminant minimization. It entails de-proteinization by centrifugal ultra filtration through ultracel 3 K filter and analysis by high performance anion exchange liquid chromatography (Sphereclone, 5 μ SAX column with UV detection at 214 nm. The lower detection limit of the assay procedure is 50 pmoles in all midgut and haemolymph samples. Retention times for NO2- and NO3- in standards and in mid-gut samples were 3.42 and 4.53 min. respectively. Assay linearity for standards ranged between 50 nM and 1 mM. Recoveries of NO2- and NO3- from spiked samples (1–100 μM and from the extracted standards (1–100 μM were calculated to be 100%. Intra-assay and inter assay variations and relative standard deviations (RSDs for NO2- and NO3- in spiked and un-spiked midgut samples were 5.7% or less. Increased levels NO2- and NO3- in midguts and haemolymph of An. culicifacies sibling species B in comparison to species A reflect towards a mechanism of refractoriness based on AcNOS physiology. Conclusion HPLC is a sensitive

  17. Viability of microflora pf market fermented milk products in simulated conditions of gastric and duodenum

    Directory of Open Access Journals (Sweden)

    Małgorzata Ziarno

    2010-06-01

    Full Text Available Background. The non-probiotic lactic acid bacteria have only rarely been used in in vitro or in vivo studies, because they are not considered to exert health benefits. To exert the beneficial effect in human organism, LAB needs to meet some criteria, for example the viability in the gastrointestinal tract (GIT. The aim of this work was to determine the viability of microflora of chosen market non-probiotic fermented milk products in simulated gastric and duodenal fluids. Material and methods. Ten market non-probiotic fermented milk products bought in Warsaw were used in this study. Ten grams of each product have been suspended in simulated gastric fluid, and then the mixture has been transferred into simulated duodenal fluid. Immediately after bacteria inoculums addition and at the end of the experiment, the number of lactobacilli and lactococci was measured. Results. The number of lactococci or streptococci decreased by 0.1-0.3 log cycle after 3 h in gastric mixture. Only in one yoghurt the population of streptococci decreased by 0.9 log cycle. The population of lactobacilli did not change in condition of simulated gastric fluid. The significant reduction of lactobacilli, lactococci and streptococci population was observed after the transfer of mixture into simulated duodenal fluid and incubation in this condition for 5 h. After the end of experiments in every studied sample the number of microflora remained at the level above 6 log CFU/mL. Conclusions. The results indicate that the condition simulating gastric fluid is not a menace to viability of lactic acid bacteria, if they are protected by milk products. The significant reduction of bacteria number in simulated duodenal fluid was probably caused by cell shock to intensive pH change of environment and loss of protective barrier caused by digestive enzymes activity.

  18. Effect of smoking on subgingival microflora of patients with periodontitis in Japan

    Directory of Open Access Journals (Sweden)

    Yamada Satoru

    2011-01-01

    Full Text Available Abstract Background Smoking is a risk factor for periodontitis. To clarify the contribution of smoking to periodontitis, it is essential to assess the relationship between smoking and the subgingival microflora. The aim of this study was to gain an insight into the influence of smoking on the microflora of Japanese patients with periodontitis. Methods Sixty-seven Japanese patients with chronic periodontitis (19 to 83 years old, 23 women and 44 men were enrolled in the present study. They consisted of 30 smokers and 37 non-smokers. Periodontal parameters including probing pocket depth (PPD and bleeding on probing (BOP and oral hygiene status were recorded. Detection of Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia, Tannerella forsythia, Fusobacterium nucleatum/periodonticum, Treponema denticola and Campylobacter rectus in subgingival plaque samples was performed by polymerase chain reaction. Association between the detection of periodontopathic bacteria and smoking status was analyzed by multiple logistic regression analysis and chi-square test. Results A statistically significant association was found between having a PPD ≥ 4 mm and detection of T. denticola, P. intermedia, T. forsythia, or C. rectus, with odds ratios ranging from 2.17 to 3.54. A significant association was noted between BOP and the detection of C. rectus or P. intermedia, and smoking, with odds ratios ranging from 1.99 to 5.62. Prevalence of C. rectus was higher in smokers than non-smokers, whereas that of A. actinomycetemcomitans was lower in smokers. Conclusions Within limits, the analysis of the subgingival microbial flora in smokers and non-smokers with chronic periodontitis suggests a relevant association between smoking and colonization by the specific periodontal pathogens including C. rectus.

  19. Pharyngeal microflora disruption by antibiotics promotes airway hyperresponsiveness after respiratory syncytial virus infection.

    Directory of Open Access Journals (Sweden)

    Ke Ni

    Full Text Available BACKGROUND: Regulatory T cells (Treg cells, which are essential for regulation of immune response to respiratory syncytial virus (RSV infection, are promoted by pharyngeal commensal pneumococcus. The effects of pharyngeal microflora disruption by antibiotics on airway responsiveness and relative immune responses after RSV infection have not been clarified. METHODS: Female BALB/c mice (aged 3 weeks were infected with RSV and then treated with either oral antibiotics or oral double distilled water (ddH(2O from 1 d post infection (pi. Changes in pharyngeal microflora were analyzed after antibiotic treatment for 7 d and 14 d. At 8 d pi and 15 d pi, the inflammatory cells in bronchoalveolar lavage fluid (BALF were investigated in combination with tests of pulmonary histopathology, airway hyperresponsiveness (AHR, pulmonary and splenic Treg cells responses. Pulmonary Foxp3 mRNA expression, IL-10 and TGF-β1 in BALF and lung homogenate were investigated at 15 d pi. Ovalbumin (OVA challenge was used to induce AHR after RSV infection. RESULTS: The predominant pharyngeal commensal, Streptococcus, was cleared by antibiotic treatment for 7 d. Same change also existed after antibiotic treatment for 14 d. After RSV infection, AHR was promoted by antibiotic treatment at 15 d pi. Synchronous decreases of pulmonary Treg cells, Foxp3 mRNA and TGF-β1 were detected. Similar results were observed under OVA challenge. CONCLUSIONS: After RSV infection, antibiotic treatment cleared pharyngeal commensal bacteria such as Streptococcus, which consequently, might induce AHR and decrease pulmonary Treg cells.

  20. Changes in pharyngeal aerobic microflora in oral breathers after palatal rapid expansion

    Directory of Open Access Journals (Sweden)

    Ripa Antonio

    2006-01-01

    Full Text Available Abstract Background The purpose of this study was to investigate in oral breathing children the qualitative and quantitative effects on aerobic and facultatively anaerobic oropharyngeal microflora of respiratory function improved by rapid palatal expansion (RPE. Methods In an open clinical trial, we studied 50 oral breathers, aged 8 to 14 years and suffering from both maxillary constriction and posterior cross-bite. At baseline, patients were examined by a single otorhinolaryngologist (ENT, confirming nasal obstruction in all subjects by posterior rhino-manometric test. Patients were evaluated three times by oropharyngeal swabs:1 at baseline (T = 0; 2 after palatal spreading out (T = 1; and 3 at the end of RPE treatment (T = 2. With regard to the microbiological aspect, the most common and potentially pathogenic oral microrganisms (i.e. Streptococcus pyogenes, Diplococcus pneumoniae, Staphylococcus aureus, Haemophilus spp, Branhamella catarrhalis, Klebsiella pneumoniae, Candida albicans were specifically detected in proper culture plates, isolated colonies were identified by means of biochemical tests and counted by calibrated loop. The data were analyzed by means of the following tests: Chi-square test, Fisher's exact test and Wilcoxon's test. Results After the use of RME there was a statistically significant decrease of Staphylococcus aureus stock at CFU/mLat T1(P = 0.0005; Z = -3,455 by Wilcoxon Rank test and T2 (P Conclusion Our data suggest that RPE therapy in oral breathers may strongly reduce the pathogenic aerobic and facultatively anaerobic microflora in the oral pharynx after a normalization of the upper airways function, and may reduce the risk of respiratory infections.

  1. Gene cloning and expression of cadherin in midgut of Helicoverpa armigera and its Cry1A binding region

    Institute of Scientific and Technical Information of China (English)

    WANG Guirong; WU Kongming; LIANG Gemei; GUO Yuyuan

    2005-01-01

    Cadherins belong to one of the families of animal glycoproteins responsible for calcium-dependent cell-cell adhesion. Recent literatures showed that the cadherin-like in midgut of several insects served as the receptor of Bt toxin Cry1A and the variation of cadherin-like is related to insect's resistance to Cry1A. The full-length cDNA encoding cadherin-like of Helicoverpa armigera is cloned by degenerate PCR and RACE techniques and the gene was designated as BtR-harm, which is 5581 bp in full-length, encoding 1730 amino acid residues (BtR-harm was deposited in GenBank and the accession number is AF519180). Its predicted molecular weight and isoelectric point were 195.39 kDa and 4.23, respectively. The inferred amino acid sequence includes a signal sequence, 11 cadherin repeats, a membrane-proximal region, a transmembrane region and a cytoplasmic region. Sequence analysis indicated that the deduced protein sequence was most similar to the cadherin-like from Heliothis virescens with 84.2% identity and highly similar to three other lepidopteran cadherin from Bombyx mori, Manduca sexta and Pectinophora gossypiella, with the sequence identities of 60.3.6%, 57.5% and 51.0%, respectively. The cDNA encoding cadherin gene was expressed successfully in E. coli and the recombinant proteins can bind with Cry1Ac. Truncation analysis and binding experiment of BtR-harm revealed that the Cry1A binding region was a contiguous 244-amino acid sequence, which located between amino acid 1217 and 1461. Semi-quantitative RT-PCR analysis showed that BtR-harm was highly expressed in midgut of H. armigera, very low expressed in foregut and hindgut and was not expressed in other tissues. After H. armigera producing resistance to Cry1Ac, the expression quantity of BtR-harm significantly decreased in midgut of H. armigera. It is the first confirmation that BtR-harm can function as receptor of Cry1Ac in H. armigera and the binding region was located on a contiguous 244 amino acid sequence

  2. Active site characterization and molecular cloning of Tenebrio molitor midgut trehalase and comments on their insect homologs.

    Science.gov (United States)

    Gomez, Ana; Cardoso, Christiane; Genta, Fernando A; Terra, Walter R; Ferreira, Clélia

    2013-08-01

    The soluble midgut trehalase from Tenebrio molitor (TmTre1) was purified after several chromatographic steps, resulting in an enzyme with 58 kDa and pH optimum 5.3 (ionizing active groups in the free enzyme: pK(e1) = 3.8 ± 0.2 pK(e2) = 7.4 ± 0.2). The purified enzyme corresponds to the deduced amino acid sequence of a cloned cDNA (TmTre1-cDNA), because a single cDNA coding a soluble trehalase was found in the T. molitor midgut transcriptome. Furthermore, the mass of the protein predicted to be coded by TmTre1-cDNA agrees with that of the purified enzyme. TmTre1 has the essential catalytic groups Asp 315 and Glu 513 and the essential Arg residues R164, R217, R282. Carbodiimide inactivation of the purified enzyme at different pH values reveals an essential carboxyl group with pKa = 3.5 ± 0.3. Phenylglyoxal modified a single Arg residue with pKa = 7.5 ± 0.2, as observed in the soluble trehalase from Spodoptera frugiperda (SfTre1). Diethylpyrocarbonate modified a His residue that resulted in a less active enzyme with pK(e1) changed to 4.8 ± 0.2. In TmTre1 the modified His residue (putatively His 336) is more exposed than the His modified in SfTre1 (putatively His 210) and that affects the ionization of an Arg residue. The architecture of the active site of TmTre1 and SfTre1 is different, as shown by multiple inhibition analysis, the meaning of which demands further research. Trehalase sequences obtained from midgut transcriptomes (pyrosequencing and Illumina data) from 8 insects pertaining to 5 different orders were used in a cladogram, together with other representative sequences. The data suggest that the trehalase gene went duplication and divergence prior to the separation of the paraneopteran and holometabolan orders and that the soluble trehalase derived from the membrane-bound one by losing the C-terminal transmembrane loop.

  3. Deletion of the NSm virulence gene of Rift Valley fever virus inhibits virus replication in and dissemination from the midgut of Aedes aegypti mosquitoes.

    Directory of Open Access Journals (Sweden)

    Rebekah C Kading

    2014-02-01

    Full Text Available BACKGROUND: Previously, we investigated the role of the Rift Valley fever virus (RVFV virulence genes NSs and NSm in mosquitoes and demonstrated that deletion of NSm significantly reduced the infection, dissemination, and transmission rates of RVFV in Aedes aegypti mosquitoes. The specific aim of this study was to further characterize midgut infection and escape barriers of RVFV in Ae. aegypti infected with reverse genetics-generated wild type RVFV (rRVF-wt or RVFV lacking the NSm virulence gene (rRVF-ΔNSm by examining sagittal sections of infected mosquitoes for viral antigen at various time points post-infection. METHODOLOGY AND PRINCIPAL FINDINGS: Ae. aegypti mosquitoes were fed an infectious blood meal containing either rRVF-wt or rRVF-ΔNSm. On days 0, 1, 2, 3, 4, 6, 8, 10, 12, and 14 post-infection, mosquitoes from each experimental group were fixed in 4% paraformaldehyde, paraffin-embedded, sectioned, and examined for RVFV antigen by immunofluorescence assay. Remaining mosquitoes at day 14 were assayed for infection, dissemination, and transmission. Disseminated infections were observed in mosquitoes as early as three days post infection for both virus strains. However, infection rates for rRVF-ΔNSm were statistically significantly less than for rRVF-wt. Posterior midgut infections in mosquitoes infected with rRVF-wt were extensive, whereas midgut infections of mosquitoes infected with rRVF-ΔNSm were confined to one or a few small foci. CONCLUSIONS/SIGNIFICANCE: Deletion of NSm resulted in the reduced ability of RVFV to enter, replicate, and disseminate from the midgut epithelial cells. NSm appears to have a functional role in the vector competence of mosquitoes for RVFV at the level of the midgut barrier.

  4. Molecular methods to evaluate effects of feed additives and nutrients in poultry gut microflora Metodologias moleculares para avaliar efeitos de aditivos e nutrientes na microflora intestinal das aves

    Directory of Open Access Journals (Sweden)

    Edgar O. Oviedo-Rondón

    2009-07-01

    Full Text Available Intestines of each animal are the niche of a complex and dynamic ecosystem with important effects to the host. The members or final products of this ecosystem influence nutrient digestion, absorption, mucosa metabolism, general physiology, and local and systemic immunological responses of avian hosts. Better understanding of the avian gut microbial ecosystem may lead to improvements on poultry productivity, health, welfare, and reduction of food borne pathogens and the environmental impact of poultry production for a more sustainable industry. Molecular methods of microbial ecology are key tools to gain this knowledge. The objective of this presentation is to outline the basic concepts, applications, advantages, limitations, and evolution of these molecular methods used to study intestinal microbial ecology. The final goal is to stimulate their application in poultry applied research and development of new feed additives. Some practical examples in poultry research will be described to illustrate their relevance to advance in control methods for pathogens, avoid or manage disbiosis or subclinical intestinal diseases, reduce environmental impact, elucidate effects of nutrients in gut mucosa, microflora, and in general to improve poultry performance.O intestino de cada animal é o nicho de um ecossistema complexo e dinâmico com efeitos importantes para o hospedeiro. As comunidades microbianas componentes deste ecossistema e/ou os produtos finais do metabolismo influenciam a digestão e absorção de nutrientes, o metabolismo das mucosas, a fisiologia geral e as respostas imunitárias locais e gerais da ave hospedeira. A melhor compreensão do ecossistema microbiano do intestino das aves pode levar a melhorias na produtividade, saúde, bem estar, e redução de agente patogênicos dos alimentos e do impacto ambiental da produção avícola para uma indústria mais sustentável. Os métodos moleculares da ecologia microbiana são ferramentas chaves

  5. Changes in midgut endopeptidase activity of Spodoptera frugiperda (Lepidoptera: Noctuidae) are responsible for adaptation to soybean proteinase inhibitors.

    Science.gov (United States)

    Paulillo, L C; Lopes, A R; Cristofoletti, P T; Parra, J R; Terra, W R; Silva-Filho, M C

    2000-06-01

    The development of transgenic maize plants expressing soybean proteinase inhibitors could reduce the economic damage of one of the major maize pests in Brazil, the fall armyworm, Spodoptera frugiperda (J.E. Smith, 1797). We examined the influence of soybean proteinase inhibitors on digestive enzyme properties and development of S. frugiperda larvae. The inhibition of trypsin and chymotrypsin activities in vitro by soybean proteinase inhibitors suggested that either Kunitz (SBTI) or Bowman-Birk (SBBI) would have a potential antimetabolic effect when ingested by insect larvae. However, chronic ingestion of semipurified soybean inhibitors did not result in a significant reduction of growth and development of fall armyworm. Therefore, digestive serine proteinase activities (trypsin and chymotrypsin) of fall armyworm larvae were characterized. The results suggest that S. frugiperda was able to physiologically adapt to dietary proteinase inhibitors by altering the complement of proteolytic enzymes in the insect midguts.

  6. Separation of Binding Protein of Celangulin V from the Midgut of Mythimna separata Walker by Affinity Chromatography

    Directory of Open Access Journals (Sweden)

    Lina Lu

    2015-05-01

    Full Text Available Celangulin V, an insecticidal compound isolated from the root bark of Chinese bittersweet, can affect the digestive system of insects. However, the mechanism of how Celangulin V induces a series of symptoms is still unknown. In this study, affinity chromatography was conducted through coupling of Celangulin V-6-aminoacetic acid ester to the CNBr-activated Sepharose 4B. SDS-PAGE was used to analyze the collected fraction eluted by Celangulin V. Eight binding proteins (Zinc finger protein, Thioredoxin peroxidase (TPx, Glyceraldehyde 3-phosphate dehydrogenase (GAPDH, SUMO E3 ligase RanBP2, Transmembrane protein 1, Actin, APN and V-ATPase were obtained and identified by LC/Q-TOF-MS from the midgut of Mythimna separata larvae. The potential of these proteins to serve as target proteins involved in the insecticidal activity of Celangulin V is discussed.

  7. Characterization of the proteases in the midgut of the xylophagous larvae of Oemona hirta (Coleoptera:Cerambycidae)

    Institute of Scientific and Technical Information of China (English)

    Brian David Shaw; John Tane Christeller

    2009-01-01

    The protein digestive capability oftbe larvae of the longhorn beetle (Oemona hirta,Coleoptera:Cerambycidae,Fabricius,1775) was investigated.This species feeds only on wood where there is a high proportion of vascular tissue.The pH of the midgut,the major digestive organ,was alkaline and protein hydrolysis was maximal at alkaline pH.Use of specific synthetic peptide substrates showed that the major protease activities were the endopeptidases,trypsin and chymotrypsin-like activity,and the exopeptidase,leucine aminopeptidase and the pH curves corresponded to that with protein substrate.Studies using a range ofsefine protease inhibitors as well as specific inhibitors ofmetalloproteases,cysteine proteases and aspartate proteases confirmed a serine protease-based digestive system similar to earlier reports of sapwood-feeding Cerambycids.Control of these insect pests using protease inhibitors is discussed.

  8. Deltamethrin-Mediated Toxicity and Cytomorphological Changes in the Midgut and Nervous System of the Mayfly Callibaetis radiatus

    Science.gov (United States)

    Gutiérrez, Yeisson; Santos, Helen P.; Serrão, José Eduardo; Oliveira, Eugênio E.

    2016-01-01

    Immature instars of mayflies are important constituents of the food web in aquatic ecosystems (especially in Neotropical regions) and they are among the most susceptible arthropods to pyrethroid insecticides. These insecticides have been recognized as important stressors of freshwater ecosystems, but their cellular effects in aquatic insects have been neglected. Here, we assessed the susceptibility to deltamethrin (a typical type II pyrethroid) as well as the deltamethrin-mediated cytomorphological changes in the central nervous system and midgut of the mayfly Callibaetis radiatus. While the deltamethrin LC50 for 24h of exposure was of 0.60 (0.46–0.78) μg of a.i/L, the survival of C. radiatus was significantly reduced in deltamethrin concentrations ≥ 0.25 μg a.i/L at 96h of exposure. Sub-lethal deltamethrin exposure severely affected the cytomorphology of C. radiatus midgut (e.g., muscle layer retraction, cytoplasm vacuolation, nucleus and striated border disorganization) and also induced slight cytomorphological changes in the brain (e.g., presence of pyknotic nuclei) and in the thoracic ganglia (e.g., vacuolation of neurons and presence of pyknotic nuclei) of these insects. However, DNA damage was absent in all of these organs, suggesting that the sublethal cellular stress induced by deltamethrin might disrupt physiological processes (e.g., metabolism or electrical signal transmission) rather than cause cell death (e.g., apoptosis) in C. radiatus. Thus, our findings indicated that deltamethrin actions at cellular levels represent a clear indication of sublethal effects on the C. radiatus survival abilities. PMID:27031238

  9. Variant vicilins from a resistant Vigna unguiculata lineage (IT81D-1053) accumulate inside Callosobruchus maculatus larval midgut epithelium.

    Science.gov (United States)

    Oliveira, Gabriel B; Kunz, Daniele; Peres, Tanara V; Leal, Rodrigo B; Uchôa, Adriana F; Samuels, Richard I; Macedo, Maria Lígia R; Carlini, Célia R; Ribeiro, Alberto F; Grangeiro, Thalles B; Terra, Walter R; Xavier-Filho, José; Silva, Carlos P

    2014-02-01

    It has been demonstrated that variant vicilins are the main resistance factor of cowpea seeds (Vigna unguiculata) against attack by the cowpea beetle Callosobruchus maculatus. There is evidence that the toxic properties of these storage proteins may be related to their interaction with glycoproteins and other microvillar membrane constituents along the digestive tract of the larvae. New findings have shown that following interaction with the microvilli, the vicilins are absorbed across the intestinal epithelium and thus reach the internal environment of the larvae. In the present paper we studied the insecticidal activity of the variant vicilins purified from a resistant cowpea variety (IT81D-1053). Bioassays showed that the seeds of this genotype affected larval growth, causing developmental retardation and 100% mortality. By feeding C. maculatus larvae on susceptible and IT81D-1053 derived vicilins (FITC labelled or unlabelled), followed by fluorescence and immunogold cytolocalization, we were able to demonstrate that both susceptible and variant forms are internalized in the midgut cells and migrate inside vesicular structures from the apex to the basal portion of the enterocytes. However, when larvae were fed with the labelled vicilins for 24h and then returned to a control diet, the concentration of the variant form remained relatively high, suggesting that variant vicilins are not removed from the cells at the same rate as the non-variant vicilins. We suggest that the toxic effects of variant vicilins on midgut cells involve the binding of these proteins to the cell surface followed by internalization and interference with the normal physiology of the enterocytes, thereby affecting larval development in vivo.

  10. Deltamethrin-Mediated Toxicity and Cytomorphological Changes in the Midgut and Nervous System of the Mayfly Callibaetis radiatus.

    Directory of Open Access Journals (Sweden)

    Yeisson Gutiérrez

    Full Text Available Immature instars of mayflies are important constituents of the food web in aquatic ecosystems (especially in Neotropical regions and they are among the most susceptible arthropods to pyrethroid insecticides. These insecticides have been recognized as important stressors of freshwater ecosystems, but their cellular effects in aquatic insects have been neglected. Here, we assessed the susceptibility to deltamethrin (a typical type II pyrethroid as well as the deltamethrin-mediated cytomorphological changes in the central nervous system and midgut of the mayfly Callibaetis radiatus. While the deltamethrin LC50 for 24 h of exposure was of 0.60 (0.46-0.78 μg of a.i/L, the survival of C. radiatus was significantly reduced in deltamethrin concentrations ≥ 0.25 μg a.i/L at 96 h of exposure. Sub-lethal deltamethrin exposure severely affected the cytomorphology of C. radiatus midgut (e.g., muscle layer retraction, cytoplasm vacuolation, nucleus and striated border disorganization and also induced slight cytomorphological changes in the brain (e.g., presence of pyknotic nuclei and in the thoracic ganglia (e.g., vacuolation of neurons and presence of pyknotic nuclei of these insects. However, DNA damage was absent in all of these organs, suggesting that the sublethal cellular stress induced by deltamethrin might disrupt physiological processes (e.g., metabolism or electrical signal transmission rather than cause cell death (e.g., apoptosis in C. radiatus. Thus, our findings indicated that deltamethrin actions at cellular levels represent a clear indication of sublethal effects on the C. radiatus survival abilities.

  11. Entry into Midgut Epithelial Cells is a Key Step in the Selection of Genotypes in a Nucleopolyhedrovirus

    Institute of Scientific and Technical Information of China (English)

    Gabriel Clavijo; Trevor Williams; Delia Mu(n)oz; Miguel L(o)pez-Ferber; Primitivo Caballero

    2009-01-01

    An isolate of the Spodoptera frugiperda multiple nucleopolyhedrovirus comprises a stable proportion of deletion genotypes (e.g., SfNIC-C), that lack pif1 and pif2 rendering them noninfectious per os, and that survive by complementation with a complete genotype (SfNIC-B) in coinfected cells. To determine whether selection for particular ratios of complete and deletion genotypes occurs mainly during the establishment of the primary infection in insect midgut cells or during subsequent systemic infection, we examined genotype frequencies in insects that fed on OBs comprising different co-occluded mixtures of genotypes. Dramatic changes in genotype frequencies were observed between the OB inoculum and budded virus (BV) samples taken from larvae inoculated with OBs comprising 10% SfNIC-B + 90% SfNIC-C indicating that a marked reduction of SfNIC-C genotype had occurred in the insect midgut due to the immediate elimination of all OBs that originated from cells that had been infected only by SfNIC-C. In contrast, immediate changes were not observed in OBs comprising mixtures of 50% SfNIC-B + 50% SfNIC-C or those comprising 10% SfNIC-B + 90% SfNIC-C as most of the OBs in these mixtures originated from cells that had been infected by both genotypes. Subsequent changes in genotypic frequencies during five days of systemic infection were fairly small in magnitude for all genotypic mixtures. We conclude that the prevalence of defective genotypes in the SfNIC population is likely determined by a balance between host selection against OBs produced in cells infected by SfNIC-C alone and within-host selection for fast-replicating deletion genotypes. The strength of intra-host selection is likely modulated by changes in MOI during the infection period.

  12. [Content of parodontopathogenic microflora in cases of parodontitis of different severity degrees according to the data of polymerase chain reaction method].

    Science.gov (United States)

    Grudianov, A I; Ovchinnikova, V V

    2008-01-01

    The use of semiquantitative determination of parodontopathogenic microflora of parodontal pockets (PP) by polymerase chain reaction (PCR) in cases of chronic generalized parodontitis of different severity degrees was investigated. 63 patients were examined - 6 with light parodontitis, 16 with parodontitis of middle severity and 41 with heavy parodontitis. The content of PP was studied. DNA of the main representatives of parodontopathogenic microflora was determined by PCR method.

  13. Efectos del daño de la mosca del olivo y del atroje sobre la microflora en pasta y la acidez del aceite virgen de oliva

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    Martínez, J. A.

    2003-09-01

    Full Text Available The effects of olive fly damage, olive storage, harvest date, olive grove plot and their interactions on the microflora associated with olive paste and oil acidity were studied. Microflora and oil acidity were significantly affected by the four studied factors and by several interactions between them. Fly damage and olive storage interacted sinergically increasing oil acidity. The relationship between fly damage and oil acidity was lineal although sometimes it was not significant depending on microflora populations. The relationship between microflora populations and oil acidity fitted to a logarithmic model. Major microorganisms in olive paste were bacteria (Xanthomonas, yeasts (mostly Torulopsis and Candida and in a smaller measure moulds (mainly Fusarium and Penicillium. Results overall suggest that a qualitative damage threshold based on the percentage of damaged fruits in order to infer oil acidity may be unfeasible in most instances.Se estudiaron los efectos del daño de mosca, atroje, fecha de recolección, parcela de olivo y de sus interacciones, sobre la microflora asociada a la pasta de aceituna y la acidez del aceite obtenido. Microflora y acidez se vieron significativamente afectados por los cuatro factores estudiados y por varias de sus interacciones. Daño y atroje interactuaron sinérgicamente para incrementar la acidez. La relación entre el daño de mosca y la acidez fue lineal aunque a veces no fue significativa dependiendo de la microflora. La relación entre la microflora y la acidez se ajustó a un modelo logarítmico. Los microorganismos predominantes en la pasta fueron bacterias (Xanthomonas, levaduras (fundamentalmente Torulopsis y Candida y en menor medida otros hongos (principalmente Fusarium y Penicillium. Los resultados sugieren que un umbral de daño cualitativo basado en el porcentaje de frutos picados para inferir la acidez del aceite es de difícil implementación.

  14. Effect of the gastrointestinal microflora on induction and maintenance of oral tolerance to ovalbumin in C3H/HeJ mice.

    OpenAIRE

    Moreau, M C; Corthier, G

    1988-01-01

    The effect of the digestive microflora on oral tolerance to ovalbumin was studied by using axenic (germfree) and conventional C3H/HeJ mice. In contrast to reported results of studies with sheep erythrocytes, oral administration of ovalbumin induced tolerance in axenic mice, but the maintenance of tolerance was found to be of shorter duration than was with conventional mice. These data indicate that the contribution of the microflora to oral tolerance depends on the antigen used.

  15. ANTICOMPLEMENTARY ACTIVITY OF THE MICROFLORA OF THE INTESTINE IN PEOPLE WHO ARE OVERWEIGHT AND CARDIOVASCULAR PATHOLOGY

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    Osolodchenko TP

    2015-04-01

    of cases. Thus, 66,67 % of lactobacilli and 68,59 % of bifidobacteria did not have the ability to inactivate the compliment, but 33,33 % of lactobacilli and 31,48 % of bifidobacteria were able to inactivate the complement at the concentration of 5 hem.unit/mL. The percentage of strains that inactivated the complement at the concentration of 10 hem.unit/mL was 20,83 % of lactobacilli and 11,1 % of bifidobacteria. ACA was absent in 80,95 % of lactobacilli from people with obesity without comorbidity. In obese hypertension patients this percentage was 56,25 %, and if the patient had IHD, the percentage decreased to 14,29 %. The percentage of bifidobacteria representatives which did not have the ability to inactivate the complement taken from people without cardiovascular pathology was 76,92 %. In people suffering from hypertensive disease, it dropped slightly to 58,82 %, and if people had IHD, it increased to 81,82%. Conclusion. 1. The representatives of intestinal microflora from people with different weight have different ability to inactivate the complement. The most likely reason for it is the changing nature of modern human nutrition. 2. Gram-negative intestinal microflora has the most distinct ability to inactivate the complement, which may indicate its role in the development of chronic inflammation of low-grade activity in patients with obesity. 3. Naturally, intestinal microflora in a small percentage (33,33 % of lacto- and 31,48 % of bifidobacteria has the ability to inactivate the compliment. Increase of ACA of lactobacilli and decrease of ACA of bifidobacteria from patients with concomitant cardiovascular pathology can be explained by the influence of comorbidity on the biological properties of microflora as well as taking a large number of medicine by this group of patients for the treatment of hypertensive disease and ischemic heart disease, the influence of which on normal intestinal microflora is not sufficiently studied at present and requires a further

  16. Noni (Morinda citrifolia L.) Fruit Extracts Improve Colon Microflora and Exert Anti-Inflammatory Activities in Caco-2 Cells.

    Science.gov (United States)

    Huang, Hsin-Lun; Liu, Cheng-Tzu; Chou, Ming-Chih; Ko, Chien-Hui; Wang, Chin-Kun

    2015-06-01

    Intestinal microflora and inflammation are associated with the risk of inflammatory bowel diseases. Noni (Morinda citrifolia L.) has various bioactivities, but its effect on colon health remains unknown. This study focused on the effects of fermented noni fruit extracts on colon microflora and inflammation of colon epithelial cells. The anti-inflammatory activities of ethanol and ethyl acetate extracts on Caco-2 cells were evaluated including interleukin-8 (IL-8) and cyclooxygenase-2 (COX-2). The growth of Lactobacillus and Bifidobacterium species was promoted by ethanol extract. Ethyl acetate extract decreased intracellular reactive oxygen species and significantly suppressed COX-2, IL-8, and prostaglandin E2 production and neutrophil chemotaxis by suppressing the translocation of the p65 subunit. Quercetin was the main contributor to the anti-inflammatory activity. The fermented noni fruit promoted probiotic growths and downregulated the intracellular oxidation and inflammation in Caco-2 cells. These results suggest that fermented noni fruit might protect against inflammatory diseases of the colon.

  17. Microbial DNA fingerprinting of human fingerprints: dynamic colonization of fingertip microflora challenges human host inferences for forensic purposes

    OpenAIRE

    Tims, S.; Wamel, van, JJ Jos; Endtz, H. P.; Belkum, van, A.; Kayser, M

    2009-01-01

    Human fingertip microflora is transferred to touched objects and may provide forensically relevant information on individual hosts, such as on geographic origins, if endogenous microbial skin species/strains would be retrievable from physical fingerprints and would carry geographically restricted DNA diversity. We tested the suitability of physical fingerprints for revealing human host information, with geographic inference as example, via microbial DNA fingerprinting. We showed that the tran...

  18. CHARACTERISTICS OF TREATMENT OF PATIENTS WITH ACUTE AND CHRONIC LARYNGITIS DEPENDING ON RESULTS OF BACTERIOLOGICAL EXAMINATION OF LARYNX MICROFLORA

    OpenAIRE

    K. I. Chuikova; T. N. Zaripova; V. I. Mukhina

    2014-01-01

    This paper describes study of larynx microflora in patients with acute laryngtis and exacerbations of chronic laryngitis. Therapeutic algorithm based on bacteriologic examination data was developed reduce. New algorithm helps reduce sickness terms and to decrease prevalence of desease.123 patients of age from 18 to 60 were under observation: 43 patients with acute laryngitis and 80 patients with exacerbation of chronic laryngitis. 22 patients with acute laryngitis and 58 patients with exacerb...

  19. Bacterial microflora isolated from the bark surface of poplars growing in areas where air pollution is very high

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    Krystyna Przybył

    2015-05-01

    Full Text Available In the autumn of 1976 bacteria of the genera Bacillus, Pseudomonas, Flavobacterium, Erwinia and Cellulomonas were isolated from the bark surface of poplars growing in protective belts around several industrial plants. It was found that the qualitative and quantitative composition of the surface bacterial microflora changes in dependence on the degree of resistance of the poplars to the action of the dust emitted by the industrial establishment and containing high amounts of heavy metals.

  20. Modelling the effect of temperature on specific death rate of the micro-flora of raw cotton wool

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    C.C. Opara

    2013-01-01

    Full Text Available Cotton wool for use in medicine must be sterilized and heat treatment is the most common sterilization technique for bulk processing of agricultural materials. Modelling the effect of temperature variations on specific death rate of the micro-flora of raw cotton wool was carried out in this project. The raw cotton wool was collected from a farmer in Kano State, the most important cotton wool producing state of Nigeria. The micro-flora was determined by inoculating and incubating with 0.85% NaCl diluents and the number of bacteria determined by plate count method. As the sterilization temperatures were varied with increased time, the thermal death rates and thermal death time of the micro-flora were determined for each temperature and these was used in the modelling. The model lnK = 83.9 – 34190(1/T shows that the sterilization temperature has a linear relationship with the death rate constant and it is in line with Arrhenius model.

  1. Non-cultural methods of human microflora evaluation for the benefit of crew medical control in confined habitat

    Science.gov (United States)

    Viacheslav, Ilyin; Lana, Moukhamedieva; Georgy, Osipov; Aleksey, Batov; Zoya, Soloviova; Robert, Mardanov; Yana, Panina; Anna, Gegenava

    2011-05-01

    Current control of human microflora is a great problem not only for the space medicine but also for practical health care. Due to many reasons its realization by classical bacteriological method is difficult in practical application or cannot be done. To evaluate non-cultural methods of microbial control of crews in a confined habitat we evaluated two different methods. The first method is based on digital treatment of microbial visual images, appearing after gram staining of microbial material from natural sample. This way the rate between gram-positive and gram-negative microbe could be gained as well as differentiation of rods and cocci could be attained, which is necessary for primary evaluation of human microbial cenosis in remote confined habitats. The other non-culture method of human microflora evaluation is gas chromatomass spectrometry (gcms) analysis of swabs gathered from different body sites. Gc-ms testing of swabs allows one to validate quantitative and special microflora based on specific lipid markers analysis.

  2. Response of soil microflora to impact of heavy metals in zones of influence of railway transport

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    N. Bobryk

    2016-03-01

    Full Text Available Monitoring research on the areas intensively and continuously affected by technogenic loading has remained topical until nowadays. The soil as a basic component of many ecosystems, including the structure of its microbial cenoses, remains an informative index of a system’s overall stability. The areas affected by railway transport have been scarcely studied. Due to the above, the aim of this work has been to establish the transformation regularities of soil microbial cenoses of territories close to railways and to establish the groups of microorganisms that are a sensitive criterion of technogenic vehicular influence. For the purpose of microbiological research, soil samples were taken at different distances from the railway track (0, 25, 50, 100 and250 m within five monitoring sections of the Tchop – Uzhhorod – Sambor railway (in the territory of Zakarpatska oblast. The number of ecological trophic groups was identified by means of inoculation on nutrient media using the method of serial dilution of soil suspensions. The research showed that in all types of soils that were adjacent to railway tracks, the number of ammonifiers and spore microbiota was high due to the high content of heavy metals (beyond the background levels. Besides, the bacterial microflora on beef-extract agar was characterized by homogeneity with domination of enteric bacteria and spore bacteria. Simultaneously, the numbers of nitrogen-fixing microorganisms, micromycetes, oligonitrophils, amylolytic and pedotrophic microflorae were shown to be low compared to the control. Farther from the railway track, pigmental species of bacteria appeared in the soil samples, attesting to the activity of self-purification processes. Correlation analysis of the data showed that the soil microbiota of the railway-side areas was undergoing changes as affected by heightened contents of heavy metals. Existence of medium and close connections was established between the number of

  3. Antimicrobial resistance in the subgingival microflora in patients with adult periodontitis - A comparison between The Netherlands and Spain

    NARCIS (Netherlands)

    van Winkelhoff, AJ; Gonzales, DH; Winkel, EG; Dellemijn-Kippuw, N; Vandenbroucke-Grauls, CMJE; Sanz, M

    2000-01-01

    Background: The widespread use of antibiotics for prophylaxis and treatment of bacterial infections has lead to the emergence of resistant human pathogens. Great differences have been documented between European countries in the use of systemic antibiotics. In parallel, significant differences in le

  4. Using soil health to assess ecotoxicological impacts of pollutants on soil microflora.

    Science.gov (United States)

    Bécaert, Valérie; Deschênes, Louise

    2006-01-01

    Microorganisms are essential for a properly functioning soil ecosystem. However, few methods allow an ecotoxicological evaluation of pollutant impact on the soil microbial community. This review proposes the use of the concept of soil health as an ecotoxicological evaluation tool for soil microflora. Initially limited to sustainable agriculture, the concept of soil health is now being applied to novel situations including contaminated and remediated soils. A large amount of work has been published in the last few decades on soil health indicators, and a review of the most relevant studies is presented here. The most cited work is that of the S-5518 committee set up in 1997 by the Soil Science Society of America (SSSA), which proposed to define soil quality as being "the capacity of a soil to function within the limits of an ecosystem, to support biological production, to maintain environmental quality and to support fauna and flora health." The soil health indicators reviewed here are the ones based on this definition because it relates well to sustainability and durability of the soil functions. Several indicators proposed in these studies could be employed in the evaluation of the ecotoxicological impact of pollutants on the soil microbial community, including microbial diversity, microbial activity, and functional stability. However, research is still required to unify the concept, to set threshold values, and to standardize methodologies.

  5. Effect of Different Oligosaccharides on Immunity, and Cecal Microflora in Broilers

    Institute of Scientific and Technical Information of China (English)

    GAO Ying; SHAN An-shan

    2004-01-01

    One hundred and forty-four Avin broilers (1 d old) were randomly divided into six treatments, with three replicates of eight birds each to investigate the effect of different oligosaccharides on index of immune organ, cecal weight and microflora in broilers. The control group (Group Ⅰ) was fed with corn-soybean meal as basal diet, and the trial groups (group Ⅱ, Ⅲ, Ⅳ, Ⅴ and Ⅵ ) were fed with basal diet plus 0.1% manoligosaccharides (MOS), 0.3% soybean-oligosaccharides (SBOS), 0.3% fructooligosaccharides (FOS), 0.3% α -glucooligosaccharides(α -GOS) and 0.03% chlortetracycline (CTC), respectively. The results showed that oligosaccharides supplementation significantly (P<0.05) increased bifidobacteria concentration ofyoung broilers and did not affect on bifidobacteria or escherichia coli concentration of the birds ( P >0.05). Cecal weights of 21 d old broilers fed with FOS and α -GOS were significantly (P <0.05) increased, higher than those with MOS and SBOS, but addition of oligosaccharides had no effect on the birds. The effect of oligosaccharides on the cecal length was the same as on the cecal weights. Oligosaccharides supplement did not affect on indexes of immune organs.

  6. QUALITATIVE EVALUATION OF SUBGINGIVAL MICROFLORA AFTER THE CHEWING OF BETEL LEAF

    Directory of Open Access Journals (Sweden)

    K Vani

    2011-04-01

    Full Text Available Betel a branching vine, scientifically called as Piper betel, is used in a number of traditional remedies and known to have immune boosting as well as antibacterial properties. This study was conducted to assess the qualitative changes in the sub-gingival micro flora, after the chewing of betel leaves, in order to evaluate the antimicrobial effect of this medicinal plant on oral pathogens.Forty volunteers were made to chew betel leaves daily for 5-10 minutes for a period of two weeks and the sub gingival plaque samples were analyzed to identify the change in type organisms if any.We have identified a wide variation in the type of organisms in the sub gingival plaque samples after the study period. Many organisms had reduced while few increased and interestingly few organisms which were not observed in the plaque sample prior to the use of betel leaf have appeared after the use. Significant alteration observed was pertaining to Streptococcus viridans group of organisms which is the commonest oral pathogen.The analysis showed chewing of betel leaf can reduce the pathogenic organisms in the sub gingival microflora.

  7. Effect of probiotics on the fecal microflora after radiotherapy: A pilot study

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    J Timko

    2013-01-01

    Full Text Available Background and Aim : The development of gastrointestinal symptoms following pelvic radiotherapy depends on morphological and functional modifications of the intestinal epithelium after radiation. The aim of this study was to evaluate and compare the effects of preventive administration of the preparation ′′5′′ Strain Dophilus and Hylak on the fecal microflora after radiotherapy in patients during radiotherapy. Materials and Methods : Fourteen patients were randomly selected and subdivided into two groups: The first group was administered ′′5′′ Strain Dophilus (L Group and the second group was administered Hylak (H Group. Radiation was delivered by a Cobalt 60 unit by using the four field box technique. The doses were divided into 2 Gy per day over 5 to 7 weeks to give the total cumulative dose of 50 Gy (2 Gy/day. High risk patients (e.g., patients with prostate cancer, received dosage 65 67 Gy (2 Gy/day. Results : Both experimental and clinical studies have shown that probiotics can effectively modulate intestinal inflammation by altering the composition and the metabolic and functional properties of gut indigenous flora. Conclusions : Many bacteria were found to be sensitive to irradiation. It would be necessary to check the possible effects of cytostatics on bacteria in larger studies.

  8. Characterisation of the microflora contaminating the wooden vats used for traditional Sicilian cheese production

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    Maria Luisa Scatassa

    2015-03-01

    Full Text Available Traditional Sicilian cheese productions are carried out employing traditional wooden vats, called tina. Many studies have highlighted the beneficial role of wooden dairy equipment by contributing to enriching the milk microflora and improving the acidification processes. The present work was undertaken to evaluate the safety of the wooden vats used to coagulate milk. To this purpose, the different microbial populations hosted onto the internal surfaces of the vats used to produce two different stretched cheeses, namely Caciocavallo Palermitano and Vastedda della valle del Belìce DOP, were investigated for the presence of spoilage and pathogenic microorganisms as well as for bacteria with inhibitory effect in vitro against pathogenic microorganisms. A wide biodiversity of protechnological lactic acid bacteria (LAB, in terms of species, was revealed. Several LAB inhibited the growth of Listeria monocytogenes ATCC 7644. The wooden vats analysed resulted safe for three main findings: absence of the main pathogenic species, presence of high levels of LAB, anti-Listeria activity of many LAB.

  9. Development of a simple bio-hydrogen production system through dark fermentation by using unique microflora

    Energy Technology Data Exchange (ETDEWEB)

    Ohnishi, Akihiro; Bando, Yukiko; Fujimoto, Naoshi; Suzuki, Masaharu [Department of Fermentation Science, Faculty of Applied Bio-Science, Tokyo University of Agriculture, 1-1 Sakuragaoka 1-chome, Setagaya-ku, Tokyo 156-8502 (Japan)

    2010-08-15

    In order to ensure efficient functioning of hydrogen fermentation systems that use Clostridium as the dominant hydrogen producer, energy-intensive process such as heat pretreatment of inoculum and/or substrate, continuous injection, and control of anaerobic conditions are required. Here, we describe a simple hydrogen fermentation system designed using microflora from leaf-litter cattle-waste compost. Hydrogen and volatile fatty acid production was measured at various hydraulic retention times, and bacterial genera were determined by PCR amplification and sequencing. Although hydrogen fermentation yield was approximately one-third of values reported in previous studies, this system requires no additional treatment and thus may be advantageous in terms of cost and operational control. Interestingly, Clostridium was absent from this system. Instead, Megasphaera elsdenii was the dominant hydrogen-producing bacterium, and lactic acid-producing bacteria (LAB) were prevalent. This study is the first to characterize M. elsdenii as a useful hydrogen producer in hydrogen fermentation systems. These results demonstrate that pretreatment is not necessary for stable hydrogen fermentation using food waste. (author)

  10. The role of intestinal microflora and probiotic bacteria in prophylactic and development of colorectal cancer

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    Ewa Wasilewska

    2013-08-01

    Full Text Available The gut microbiota comprises a large and diverse range of microorganisms whose activities have a significant impact on health. It interacts with its host at both the local and systemic level, resulting in a broad range of beneficial or detrimental outcomes for nutrition, infections, xenobiotic metabolism, and cancer. The current paper reviews research on the role of intestinal microflora in colorectal cancer development. Especially a protective effect of beneficial bacteria and probiotics on the risk of cancer development is highly discussed. There is substantial experimental evidence that the beneficial gut bacteria and their metabolism have the potential to inhibit the development and progression of neoplasia in the large intestine. Most of the data derive, however, from experimental and animal trials. Over a dozen well-documented animal studies have been published, wherein it has been clearly revealed that some lactic acid bacteria, especially lactobacilli and bifidobacteria, inhibit initiation and progression of colorectal cancer. Studies on cancer suppression in humans as a result of the consumption of probiotics are still sparse. Nevertheless, some epidemiological and interventional studies seem to confirm the bacterial anticancerogenic activity also in human gut. The mechanism by which probiotics may inhibit cancer development is unknown. Probiotics increase the amount of beneficial bacteria and decrease the pathogen level in the gut, consequently altering metabolic, enzymatic and carcinogenic activity in the intestine, decreasing inflammation and enhancing immune function, which may contribute to cancer defense.

  11. Growth performance, carcass yield and intestinal microflora populations of broilers fed diets containing thepax and yogurt

    Directory of Open Access Journals (Sweden)

    A Boostani

    2013-03-01

    Full Text Available The present study aimed at evaluating the effect of the probiotic thepax and yogurt (as probiotic on the growth response and intestinal microflora results of broiler chickens. Two hundred forty day-old Ross 308 broilers were equally distributed into 12 floor pens and reared for 42 day. The treatments consisted of yogurt (10, 5 and 2.5% during starter, grower and finisher periods in the drinking water, respectively and thepax (1000, 500, 250 g/ton-1 in the starter, grower and finisher diets, respectively, resulting three experimental diets and a control group. Each dietary treatment was fed ad-libitum to four replicate group of 20 birds at the beginning of rearing period. Birds and feed were weighed on days 21 and 42. The results of experiment indicate that diets containing feed additives improved broiler performance. The body weight gain and feed conversion ratio improved significantly more (p < 0.05 with the thepax treatment compared with the control broilers during the total rearing period. The highest (p < 0.05 carcass and thigh values were recorded for broilers fed the diet supplemented with thepax and yogurt, respectively. The lowest abdominal fat pad value was obtained in broilers fed the diet supplemented with thepax. On d 21, thepax and yogurt significantly reduced (p < 0.05 cecal Escherichia coli and Clostridium perfringens populations compared with the control group. In conclusion, thepax and yogurt improved broilers growth response and conferred intestinal health benefits to chickens by improving their microbial ecology.

  12. A Strong Impact of Genetic Background on Gut Microflora in Mice

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    R. Steven Esworthy

    2010-01-01

    Full Text Available Genetic background affects susceptibility to ileocolitis in mice deficient in two intracellular glutathione peroxidases, GPx1 and GPx2. The C57BL/6 (B6 GPx1/2 double-knockout (DKO mice have mild ileocolitis, and 129S1/Sv (129 DKO mice have severe inflammation. We used diet to modulate ileocolitis; a casein-based defined diet with AIN76A micronutrients (AIN attenuates inflammation compared to conventional LabDiets. Because luminal microbiota induce DKO ileocolitis, we assessed bacterial composition with automated ribosomal intergenic-spacer analysis (ARISA on cecal DNA. We found that mouse strain had the strongest impact on the composition of microbiota than diet and GPx genotypes. In comparing AIN and LabDiet, DKO mice were more resistant to change than the non-DKO or WT mice. However, supplementing yeast and inulin to AIN diet greatly altered microflora profiles in the DKO mice. From 129 DKO strictly, we found overgrowth of Escherichia coli. We conclude that genetic background predisposes mice to colonization of potentially pathogenic E. coli.

  13. Identification and antimicrobial resistance of microflora colonizing feral pig (Sus scrofa) of Brazilian Pantanal.

    Science.gov (United States)

    Lessa, Ss; Paes, Rcs; Santoro, Pn; Mauro, Ra; Vieira-da-Motta, O

    2011-04-01

    Antimicrobial resistance of bacteria is a worldwide problem affecting wild life by living with resistant bacteria in the environment. This study presents a discussion of outside factors environment on microflora of feral pigs (Sus scrofa) from Brazilian Pantanal. Animals had samples collected from six different body sites coming from two separated geographic areas, Nhecolandia and Rio Negro regions. With routine biochemical tests and commercial kits 516 bacteria were identified, with 240 Gram-positive, predominantly staphylococci (36) and enterococci (186) strains. Among Gram-negative (GN) bacteria the predominant specimens of Enterobacteriaceae (247) mainly represented by Serratia spp. (105), Escherichia coli (50), and Enterobacter spp. (40) and specimens not identified (7). Antimicrobial susceptibility was tested against 17 drugs by agar diffusion method. Staphylococci were negative to production of enterotoxins and TSST-1, with all strains sensitive towards four drugs and highest resistance toward ampicillin (17%). Enterococci presented the highest sensitivity against vancomycin (98%), ampicillin (94%) and tetracycline (90%), and highest resistance pattern toward oxacillin (99%), clindamycin (83%), and cotrimoxazole (54%). In GN the highest resistance was observed with Serratia marcescens against CFL (98%), AMC (66%) and AMP (60%) and all drugs was most effective against E. coli SUT, TET (100%), AMP, TOB (98%), GEN, CLO (95%), CFO, CIP (93%). The results show a new profile of oxacillin-resistant enterococci from Brazilian feral pigs and suggest a limited residue and spreading of antimicrobials in the environment, possibly because of low anthropogenic impact reflected by the drug susceptibility profile of bacteria isolated.

  14. Assessment of Fecal Microflora Changes in Pigs Supplemented with Herbal Residue and Prebiotic.

    Directory of Open Access Journals (Sweden)

    Ashis Kumar Samanta

    Full Text Available Antibiotic usage in animals as a growth promoter is considered as public health issue due to its negative impact on consumer health and environment. The present study aimed to evaluate effectiveness of herbal residue (ginger, Zingiber officinale, dried rhizome powder and prebiotic (inulin as an alternative to antibiotics by comparing fecal microflora composition using terminal restriction fragment length polymorphism. The grower pigs were offered feed containing antibiotic (tetracycline, ginger and inulin separately and un-supplemented group served as control. The study revealed significant changes in the microbial abundance based on operational taxonomic units (OTUs among the groups. Presumptive identification of organisms was established based on the fragment length of OTUs generated with three restriction enzymes (MspI, Sau3AI and BsuRI. The abundance of OTUs representing Bacteroides intestinalis, Eubacterium oxidoreducens, Selonomonas sp., Methylobacterium sp. and Denitrobacter sp. was found significantly greater in inulin supplemented pigs. Similarly, the abundance of OTUs representing Bacteroides intestinalis, Selonomonas sp., and Phascolarcobacterium faecium was found significantly greater in ginger supplemented pigs. In contrast, the abundance of OTUs representing pathogenic microorganisms Atopostipes suicloacalis and Bartonella quintana str. Toulouse was significantly reduced in ginger and inulin supplemented pigs. The OTUs were found to be clustered under two major phylotypes; ginger-inulin and control-tetracycline. Additionally, the abundance of OTUs was similar in ginger and inulin supplemented pigs. The results suggest the potential of ginger and prebioticsto replace antibiotics in the diet of grower pig.

  15. Influence of the gastrointestinal microflora and efflux transporters on the absorption of scutellarin and scutellarein.

    Science.gov (United States)

    You, Hai-Sheng; Xing, Jian-Feng; Lu, Jun; Dong, Wei-Hua; Liu, Jun-Tian; Dong, Ya-Lin

    2014-09-01

    Scutellarin (SG) and its aglycone, Scutellarein (S), are flavonoids of therapeutic cardiocerebrovascular disease. SG was hydrolyzed by bacterial enzyme into S which was absorbed in the intestine. The aim of this study was to determine the effects of the microflora in the intestinal lumen and the efflux transporter of intestinal epithelial cells on the absorption process of SG and S. After oral administration of antibiotics in Sprague-Dawley rats, the reduced bacterial enzyme formation significantly hinders the absorption of SG, whereas scarcely that of S. The absorption study in situ single-pass intestinal perfusion revealed that S could be absorbed throughout the intestine of rats. The effective intestinal permeability of S in the jejunum was much lower than in the other sections of the GI tract. The efflux transporter promoted SG secretion into lumen from enterocytes, which hindered the absorption of both SG and S into the bloodstream. The efflux transporter protein inhibitor (verapamil, probenecid and reserpine) remarkably enhanced the absorption of S and the bioconversion of S into SG in both the rat intestine and Caco-2-monolayer models.

  16. Characterisation of the Microflora Contaminating the Wooden Vats Used for Traditional Sicilian Cheese Production

    Science.gov (United States)

    Cardamone, Cinzia; Miraglia, Viviana; Lazzara, Fabrizio; Fiorenza, Gerlando; Macaluso, Giusi; Arcuri, Luigi; Settanni, Luca; Mancuso, Isabella

    2015-01-01

    Traditional Sicilian cheese productions are carried out employing traditional wooden vats, called tina. Many studies have highlighted the beneficial role of wooden dairy equipment by contributing to enriching the milk microflora and improving the acidification processes. The present work was undertaken to evaluate the safety of the wooden vats used to coagulate milk. To this purpose, the different microbial populations hosted onto the internal surfaces of the vats used to produce two different stretched cheeses, namely Caciocavallo Palermitano and Vastedda della valle del Belìce DOP, were investigated for the presence of spoilage and pathogenic microorganisms as well as for bacteria with inhibitory effect in vitro against pathogenic microorganisms. A wide biodiversity of protechnological lactic acid bacteria (LAB), in terms of species, was revealed. Several LAB inhibited the growth of Listeria monocytogenes ATCC 7644. The wooden vats analysed resulted safe for three main findings: absence of the main pathogenic species, presence of high levels of LAB, anti-Listeria activity of many LAB. PMID:27800376

  17. Application of rice rhizosphere microflora for hydrogen production from apple pomace

    Energy Technology Data Exchange (ETDEWEB)

    Doi, Tetsuya [Institute for Sustainable Agro-ecosystem Services (ISAS), Graduate School of Agriculture and Life Sciences, The University of Tokyo, Nishitokyo, Tokyo 188-0002 (Japan); Nishihara Environment Technology Inc., Tokyo 108-0023 (Japan); Matsumoto, Hisami [Nishihara Environment Technology Inc., Tokyo 108-0023 (Japan); Abe, Jun [AE-Bio, Graduate School of Agriculture and Life Sciences, The University of Tokyo, Tokyo 113-8657 (Japan); Morita, Shigenori [Institute for Sustainable Agro-ecosystem Services (ISAS), Graduate School of Agriculture and Life Sciences, The University of Tokyo, Nishitokyo, Tokyo 188-0002 (Japan)

    2010-07-15

    The combination of substrate materials and bacteria is an important factor affecting conversion technology for biological hydrogen production. We performed anaerobic hydrogen fermentation of apple pomace wastes using rhizosphere bacterial microflora of rice as the parent inoculum. In the vial test, the optimal condition for hydrogen fermentation was initial pH 6.0, 35 C, and 73.4 g pomace per liter of medium (equivalent to 10 g-hexose/L). In the batch experiment (pH 6.0, temperature 35 C) the hydrogen yield reached 2.3 mol-H{sub 2}/mol-hexose. The time course of biogas production and PCR-DGGE analysis suggest that Clostridium spp. decomposed degradable carbohydrates rapidly and a part of the refractory carbohydrate (e.g. pectin) gradually in the apple pomace slurry. In addition to hydrogen, volatile fatty acids (VFAs) were produced in the anaerobic fermentation of apple pomace, which can be a substrate for methane fermentation. The rice rhizosphere can be a promising source of inoculum bacteria for hydrogen fermentation in combination with plant material waste like apple pomace. (author)

  18. Identification and antimicrobial resistance of microflora colonizing feral pig (Sus scrofa of Brazilian Pantanal

    Directory of Open Access Journals (Sweden)

    SS Lessa

    2011-06-01

    Full Text Available Antimicrobial resistance of bacteria is a worldwide problem affecting wild life by living with resistant bacteria in the environment. This study presents a discussion of outside factors environment on microflora of feral pigs (Sus scrofa from Brazilian Pantanal. Animals had samples collected from six different body sites coming from two separated geographic areas, Nhecolandia and Rio Negro regions. With routine biochemical tests and commercial kits 516 bacteria were identified, with 240 Gram-positive, predominantly staphylococci (36 and enterococci (186 strains. Among Gram-negative (GN bacteria the predominant specimens of Enterobacteriaceae (247 mainly represented by Serratia spp. (105, Escherichia coli (50, and Enterobacter spp. (40 and specimens not identified (7. Antimicrobial susceptibility was tested against 17 drugs by agar diffusion method. Staphylococci were negative to production of enterotoxins and TSST-1, with all strains sensitive towards four drugs and highest resistance toward ampicillin (17%. Enterococci presented the highest sensitivity against vancomycin (98%, ampicillin (94% and tetracycline (90%, and highest resistance pattern toward oxacillin (99%, clindamycin (83%, and cotrimoxazole (54%. In GN the highest resistance was observed with Serratia marcescens against CFL (98%, AMC (66% and AMP (60% and all drugs was most effective against E. coli SUT, TET (100%, AMP, TOB (98%, GEN, CLO (95%, CFO, CIP (93%. The results show a new profile of oxacillin-resistant enterococci from Brazilian feral pigs and suggest a limited residue and spreading of antimicrobials in the environment, possibly because of low anthropogenic impact reflected by the drug susceptibility profile of bacteria isolated.

  19. Effect of isomalt consumption on faecal microflora and colonic metabolism in healthy volunteers.

    Science.gov (United States)

    Gostner, A; Blaut, M; Schäffer, V; Kozianowski, G; Theis, S; Klingeberg, M; Dombrowski, Y; Martin, D; Ehrhardt, S; Taras, D; Schwiertz, A; Kleessen, B; Lührs, H; Schauber, J; Dorbath, D; Menzel, T; Scheppach, W

    2006-01-01

    Due to its low digestibility in the small intestine, a major fraction of the polyol isomalt reaches the colon. However, little is known about effects on the intestinal microflora. During two 4-week periods in a double-blind, placebo-controlled, cross-over design, nineteen healthy volunteers consumed a controlled basal diet enriched with either 30 g isomalt or 30 g sucrose daily. Stools were collected at the end of each test phase and various microbiological and luminal markers were analysed. Fermentation characteristics of isomalt were also investigated in vitro. Microbiological analyses of faecal samples indicated a shift of the gut flora towards an increase of bifidobacteria following consumption of the isomalt diet compared with the sucrose diet (Pisomalt phase, the activity of bacterial beta-glucosidase decreased (Pisomalt (P=0.055). Faecal SCFA, lactate, bile acids, neutral sterols, N, NH3, phenol and p-cresol were not affected by isomalt consumption. In vitro, isomalt was metabolized in several bifidobacteria strains and yielded high butyrate concentrations. Isomalt, which is used widely as a low-glycaemic and low-energy sweetener, has to be considered a prebiotic carbohydrate that might contribute to a healthy luminal environment of the colonic mucosa.

  20. The Role of Gut Microflora and the Cholinergic Anti-inflammatory Neuroendocrine System in Diabetes Mellitus.

    Science.gov (United States)

    Parekh, Parth J; Nayi, Vipul R; Johnson, David A; Vinik, Aaron I

    2016-01-01

    The obesity epidemic has drastically impacted the state of health care in the United States. Paralleling this epidemic is the incidence of diabetes mellitus, with a notable shift toward a much younger age of onset. While central to the pathogenesis of diabetes associated with obesity is the role of inflammation attributed to "adiposopathy." Emerging data suggest that changes in sympathetic/parasympathetic balance regulated by the brain precede changes in the inflammatory cascade. It has now been established that the gut microflora contributes significantly to the activation and inhibition of autonomic control and impact the set of the neuroinflammatory inhibitory reflex mediated by the cholinergic nervous system. There has been a paradigm shift toward further investigating commensal bacteria in the pathogenesis of obesity and diabetes mellitus and its complications, as dysbiosis is thought to play a pivotal role in diabetic-associated disorders. This paper is intended to evaluate the role of intestinal dysbiosis in the pathogenesis of diabetes mellitus and examine the potential for restoration of balance via use of probiotics.

  1. The Role of Gut Microflora and the Cholinergic Anti-inflammatory Neuroendocrine System in Diabetes Mellitus

    Science.gov (United States)

    Parekh, Parth J.; Nayi, Vipul R.; Johnson, David A.; Vinik, Aaron I.

    2016-01-01

    The obesity epidemic has drastically impacted the state of health care in the United States. Paralleling this epidemic is the incidence of diabetes mellitus, with a notable shift toward a much younger age of onset. While central to the pathogenesis of diabetes associated with obesity is the role of inflammation attributed to “adiposopathy.” Emerging data suggest that changes in sympathetic/parasympathetic balance regulated by the brain precede changes in the inflammatory cascade. It has now been established that the gut microflora contributes significantly to the activation and inhibition of autonomic control and impact the set of the neuroinflammatory inhibitory reflex mediated by the cholinergic nervous system. There has been a paradigm shift toward further investigating commensal bacteria in the pathogenesis of obesity and diabetes mellitus and its complications, as dysbiosis is thought to play a pivotal role in diabetic-associated disorders. This paper is intended to evaluate the role of intestinal dysbiosis in the pathogenesis of diabetes mellitus and examine the potential for restoration of balance via use of probiotics. PMID:27375553

  2. Effects of packaging systems on the natural microflora and acceptability of chicken breast meat.

    Science.gov (United States)

    Charles, N; Williams, S K; Rodrick, G E

    2006-10-01

    The effect of 3 packaging systems on the spoilage microflora, objective color, and sensory characteristics of fresh commercial broiler chicken breast meat was investigated. Fresh skinless and boneless chicken broiler breasts were purchased from a local poultry processing plant and packaged in either (1) a conventional Styrofoam tray with polyvinyl chloride overwrap and absorbent pad, (2) a Styrofoam tray with polyvinyl chloride overwrap minus absorbent pad, or (3) a Fresh-R-Pax (FRP) container equipped with an absorbent liner-gel system. All packages were heat sealed and stored at 1.2 +/- 1 degrees C for 8 d. At each sampling period (0, 2, 4, 6, and 8 d), packages from each treatment were analyzed for Pseudomonas spp., psychrotrophic organisms, objective color, and sensory characteristics. In general, Pseudomonas spp. and psychrotrophic counts increased as storage time increased for all packaging systems. Color and overall appearance were similar (P >0.05) for all packaging systems. Although not significant, the off-odor scores for breast meat packaged in FRP were higher (P >0.05) after 6 and 8 d when compared with the breast meat packaged in a Styrofoam tray with polyvinyl chloride overwrap with or without an absorbent pad. Although the absorbent pad did not control microbial growth, it maintained aesthetic appeal by absorbing all visible moisture released from the meat during storage.

  3. [Influence of metal nanoparticles on some microorganisms and microflora of unpasteurized beer].

    Science.gov (United States)

    Pyrog, T P; Konon, A D; Antoniuk, S I; Olishevs'kyĭ, V V; Marynin, A I

    2011-01-01

    The influence of nanoparticles of gold, silver, dioxide of cerium and zirconium in low concentration (0.5-7.5 mg/l) on pure cultures of Escherichia coli IEM-1, Bacillus subtilis BT-2, Candida scottii KB-2, Saccharomyces cerevisiae OB-3, Aspergillus niger R-3, Fusarium culmorum T-7 and Penicillium chrysogenum F-7 has been investigated. Silver nanoparticles, which caused a decrease of the amount of S. cerevisiae OB-3 cells by 90% after one hour of exposition, and led to almost complete death of vegetative and spore cells of B. subtilis BT-2 after 1 and 24 h of exposition, respectively, proved to be the most effective antimicrobial preparation. The decrease by one-two orders of the amount of B. subtilis BT-2 and of S cerevisiae OB-3 cells under the influence of preparations of silver nanoparicles in the presence of gold has been established. After adding silver preparations to the unpasteurized beer one could observe a decrease by 10-20% of bacterial and by about 40% of fungi which contaminated microflora on the 20th day of shelf-life. The paper is presented in Ukrainian.

  4. Cloning of PaAtg8 and roles of autophagy in adaptation to starvation with respect to the fat body and midgut of the Americana cockroach, Periplaneta americana.

    Science.gov (United States)

    Park, Moon Soo; Takeda, Makio

    2014-05-01

    Starvation, in particular amino acid deprivation, induces autophagy in trophocytes (adipocytes), the major component of the fat body cell types, in the larvae of Drosophila melanogaster. However, the fat body of cockroach has two additional cell types: urocytes depositing uric acid in urate vacuoles as a nitrogen resource and mycetocytes harboring an endosymbiont, Blattabacterium cuenoti, which can synthesize amino acids from the metabolites of the stored uric acid. These cells might complement the roles of autophagy in recycling amino acids in the fat body or other organs of cockroaches under starvation. We investigate the presence of autophagy in tissues such as the fat body and midgut of the American cockroach, Periplaneta americana, under starvation by immunoblotting with antibody against Atg8, a ubiquitin-like protein required for the formation of autophagosomes and by electron microscopy. Corresponding changes in acid phosphatase activity were also investigated as representing lysosome activity. Starvation increased the level of an autophagic marker, Atg8-II, in both the tissues, extensively stimulating the formation of autophagic compartments in trophocytes of the fat body and columnar cells of the midgut for over 2 weeks. Acid phosphatase showed no significant increase in the fat body of the starved cockroaches but was higher in the midgut of the continuously fed animals. Thus, a distinct autophagic mechanism operates in these tissues under starvation of 2 weeks and longer. The late induction of autophagy implies exhaustion of the stored uric acid in the fat body. High activity of acid phosphatase in the midgut of the fed cockroaches might represent enhanced assimilation and not an autophagy-related function.

  5. Study of the Bacillus thuringiensis Vip3Aa16 histopathological effects and determination of its putative binding proteins in the midgut of Spodoptera littoralis

    OpenAIRE

    Abdelkefi-Mesrati, Lobna; Boukedi, Hanen; Dammak-Karray, Mariam; Sellami-Boudawara, Tahya; Jaoua, Samir; Tounsi, Slim

    2011-01-01

    The bacterium Bacillus thuringiensis produces, at the vegetative stage of its growth, Vip3A proteins with activity against a broad spectrum of lepidopteran insects. The Egyptian cotton leaf worm (Spodoptera littoralis) is an important agricultural pest that is susceptible to the Vip3Aa16 protein of Bacillus thuringiensis kurstaki strain BUPM95. The midgut histopathology of Vip3Aa fed larvae showed vacuolization of the cytoplasm, brush border membrane destruction, vesicle formation in the apic...

  6. THAP and ATF-2 regulated sterol carrier protein-2 promoter activities in the larval midgut of the yellow fever mosquito, Aedes aegypti.

    Directory of Open Access Journals (Sweden)

    Rong Peng

    Full Text Available Expression of sterol carrier protein-2 (SCP-2 in Aedes aegypti shows a distinct temporal/spatial pattern throughout the life cycle. In order to identify the transcription factors responsible for the larval temporal/spatial regulation of AeSCP-2 transcription, AeSCP-2 promoter activities were studied in vivo via transient transfection of promoter/reporter gene assays. Regulatory sequences upstream -1.3 kb of the transcription start site of AeSCP-2 were found to be critical for the in vivo temporal/spatial promoter activity. Interestingly, the -1.6 kb promoter sequence efficiently drove the larval midgut-specific siRNA expression, indicating that the -1.6 kb upstream sequence is sufficient for temporal/spatial AeSCP-2 transcriptional activity. Four transcription factors were identified in the midgut nuclear extract from feeding larvae via labeled -1.6/-1.3 kb DNA probe pull-down and proteomic analysis. Co-transfection of the promoter/reporter gene with inducible siRNA expression of each transcription factor was performed to confirm the regulatory function of individual transcription factor on AeSCP-2 transcriptional activities in the larval midgut. The results indicate that two of the identified transcription factors, Thanatos-associated protein (THAP and activating transcription factor-2 (ATF-2, antagonistically control AeSCP-2 transcriptional activity in the midgut of feeding larvae via the regulatory sequences between -1.6 to -1.3 kb 5' upstream of the transcription start site. In vivo expression knockdown of THAP and ATF-2 resulted in significant changes in developmental progression, which may be partially due to their effects on AeSCP-2 expression.

  7. Bacillus thuringiensis Cry34Ab1/Cry35Ab1 interactions with western corn rootworm midgut membrane binding sites.

    Directory of Open Access Journals (Sweden)

    Huarong Li

    Full Text Available BACKGROUND: Bacillus thuringiensis (Bt Cry34Ab1/Cry35Ab1 are binary insecticidal proteins that are co-expressed in transgenic corn hybrids for control of western corn rootworm, Diabrotica virgifera virgifera LeConte. Bt crystal (Cry proteins with limited potential for field-relevant cross-resistance are used in combination, along with non-transgenic corn refuges, as a strategy to delay development of resistant rootworm populations. Differences in insect midgut membrane binding site interactions are one line of evidence that Bt protein mechanisms of action differ and that the probability of receptor-mediated cross-resistance is low. METHODOLOGY/PRINCIPAL FINDINGS: Binding site interactions were investigated between Cry34Ab1/Cry35Ab1 and coleopteran active insecticidal proteins Cry3Aa, Cry6Aa, and Cry8Ba on western corn rootworm midgut brush border membrane vesicles (BBMV. Competitive binding of radio-labeled proteins to western corn rootworm BBMV was used as a measure of shared binding sites. Our work shows that (125I-Cry35Ab1 binds to rootworm BBMV, Cry34Ab1 enhances (125I-Cry35Ab1 specific binding, and that (125I-Cry35Ab1 with or without unlabeled Cry34Ab1 does not share binding sites with Cry3Aa, Cry6Aa, or Cry8Ba. Two primary lines of evidence presented here support the lack of shared binding sites between Cry34Ab1/Cry35Ab1 and the aforementioned proteins: 1 No competitive binding to rootworm BBMV was observed for competitor proteins when used in excess with (125I-Cry35Ab1 alone or combined with unlabeled Cry34Ab1, and 2 No competitive binding to rootworm BBMV was observed for unlabeled Cry34Ab1 and Cry35Ab1, or a combination of the two, when used in excess with (125I-Cry3Aa, or (125I-Cry8Ba. CONCLUSIONS/SIGNIFICANCE: Combining two or more insecticidal proteins active against the same target pest is one tactic to delay the onset of resistance to either protein. We conclude that Cry34Ab1/Cry35Ab1 are compatible with Cry3Aa, Cry6Aa, or Cry8Ba

  8. Long-Term Starvation and Posterior Feeding Effects on Biochemical and Physiological Responses of Midgut Gland of Cherax quadricarinatus Juveniles (Parastacidae).

    Science.gov (United States)

    Sacristán, Hernán Javier; Ansaldo, Martín; Franco-Tadic, Luis Marcelo; Fernández Gimenez, Analía Verónica; López Greco, Laura Susana

    2016-01-01

    We investigated the effect of long-term starvation and posterior feeding on energetic reserves, oxidative stress, digestive enzymes, and histology of C. quadricarinatus midgut gland. The crayfish (6.27 g) were randomly assigned to one of three feeding protocols: continuous feeding throughout 80 day, continuous starvation until 80 day, and continuous starvation throughout 50 day and then feeding for the following 30 days. Juveniles from each protocol were weighed, and sacrificed at day 15, 30, 50 or 80. The lipids, glycogen, reduced glutathione (GSH), soluble protein, lipid peroxidation (TBARS), protein oxidation (PO), catalase (CAT), lipase and proteinase activities, and histology were measured on midgut gland. Starved crayfish had a lower hepatosomatic index, number of molts, specific growth rate, lipids, glycogen, and GSH levels than fed animals at all assay times. The starvation did not affect the soluble protein, TBARS, PO levels and CAT. In starved juveniles the lipase activity decreased as starvation time increased, whereas proteinase activity decreased only at day 80. The histological analysis of the starved animals showed several signs of structural alterations. After 30 days of feeding, the starved-feeding animals exhibited a striking recovery of hepatosomatic index, number of molts, lipids and glycogen, GSH, lipase activity and midgut gland structure.

  9. The life cycle of Gregarina cuneata in the midgut of Tribolium castaneum and the effects of parasitism on the development of insects.

    Science.gov (United States)

    Gigliolli, A A S; Julio, A H F; Conte, H

    2016-04-01

    Tribolium castaneum Herbst 1797 (Coleoptera: Tenebrionidae), an important pest of stored grains and byproducts, is naturally infected by Gregarina cuneata Stein 1848 (Apicomplexa: Gregarinidae). Changes in the life cycle of insects caused by the parasite development in the midgut were studied. Trophozoites, gamonts (solitary and associated), and gametocysts were present in the midgut of the insects. In young trophozoites, the apical region differentiated into an epimerite that firmly attached the parasite to the host epithelial cells. With maturation, trophozoites developed in gamonts that were associated with the initiation of sexual reproduction in the cell cycle, culminating in the formation of the spherical gametocyst. Morpho-functional analyses indicated that gregarines absorb nutrients from infected cells and can occlude the midgut as they develop. Consequently, nutritional depletion may interfere with the host's physiology, causing decreased growth, delayed development, and high mortality rates of the parasitized insects. These results suggest G. cuneata could be an important biological agent for controlling T. castaneum in integrated pest management programs.

  10. Mesenchymal to epithelial transition during tissue homeostasis and regeneration: Patching up the Drosophila midgut epithelium.

    Science.gov (United States)

    Antonello, Zeus A; Reiff, Tobias; Dominguez, Maria

    2015-01-01

    Stem cells are responsible for preserving morphology and function of adult tissues. Stem cells divide to self-renew and to generate progenitor cells to sustain cell demand from the tissue throughout the organism's life. Unlike stem cells, the progenitor cells have limited proliferation potential but have the capacity to terminally differentiate and thereby to substitute older or damaged mature cells. Recent findings indicate that adult stem cells can adapt their division kinetics dynamically to match changes in tissue demand during homeostasis and regeneration. However, cell turnover not only requires stem cell division but also needs timed differentiation of the progenitor cells, which has been much less explored. In this Extra View article, we discuss the ability of progenitor cells to actively postpone terminal differentiation in the absence of a local demand and how tissue demand activates terminal differentiation via a conserved mesenchymal-epithelial transition program revealed in our recent EMBO J paper and other published and unpublished data. The extent of the significance of these results is discussed for models of tissue dynamics during both homeostasis and regeneration.

  11. Controle químico da microflora oral em pacientes fissurados labiopalatais durante o tratamento ortodôntico-cirúrgico: estudo piloto Oral microflora chemical control in cleft lip and palate patients during orthodontic-surgical treatment pilot study

    Directory of Open Access Journals (Sweden)

    Alcion Alves Silva

    2004-06-01

    Full Text Available O risco de infecção é uma preocupação durante o tratamento ortodôntico-cirúrgico, principalmente em pacientes como os fissurados lábio palatais, pois as condições anatômicas e oclusais contribuem para a proliferação microbiana. Métodos manuais para o controle da microflora oral são muitas vezes insuficientes para a obtenção de uma higiene satisfatória. Técnicas alternativas, como agentes químicos, têm sido propostas como procedimentos auxiliares. O objetivo deste estudo foi comparar a eficiência de dois agentes químicos como auxiliares no controle da microflora, em pacientes ortodôntico-cirúrgicos (fissurados lábio-palatais. O delineamento da pesquisa foi um estudo exploratório, microbiológico (piloto, duplo-cego, para testar os seguintes agentes químicos: triclosan 0,03% (Plax-Colgate Palmolive e clorexidina digluconada 0,12% (Duplak-Herpro/Dentsply. A amostra foi composta por 30 voluntários divididos em 3 grupos: Grupo I (controle, Grupo II (teste _ clorexidina digluconada 0,12% e Grupo III (teste _ triclosan 0,03%. Todos os grupos receberam orientação de higiene bucal prévia à primeira coleta de material. Depois de sete dias de uso dos agentes químicos uma segunda coleta foi realizada e o uso do agente foi suspenso. Uma terceira coleta foi realizada após sete dias a partir da suspensão do agente. A técnica semiquantitativa proposta por Pilonetto e Pilonetto20 foi adotada para avaliar o crescimento de colônias bacterianas. Os resultados mostraram diferenças entre a eficácia dos agentes químicos, sendo que o Grupo II mostrou uma redução mais significativa da microflora quando comparado ao grupo III.The infection risk is a preoccupation during orthodontic-surgical treatment, mainly at cleft lip and palate patients, for their anatomical and occlusal condictions, that contributs to oral microflora proliferation. Manual methods for microflora control are frequently unsuccessful to obtain a sctisfactory

  12. Exploring the midgut transcriptome of Phlebotomus papatasi: comparative analysis of expression profiles of sugar-fed, blood-fed and Leishmania major-infected sandflies

    Directory of Open Access Journals (Sweden)

    Pham Van-My

    2007-08-01

    Full Text Available Abstract Background In sandflies, the blood meal is responsible for the induction of several physiologic processes that culminate in egg development and maturation. During blood feeding, infected sandflies are also able to transmit the parasite Leishmania to a suitable host. Many blood-induced molecules play significant roles during Leishmania development in the sandfly midgut, including parasite killing within the endoperitrophic space. In this work, we randomly sequenced transcripts from three distinct high quality full-length female Phlebotomus papatasi midgut-specific cDNA libraries from sugar-fed, blood-fed and Leishmania major-infected sandflies. Furthermore, we compared the transcript expression profiles from the three different cDNA libraries by customized bioinformatics analysis and validated these findings by semi-quantitative PCR and real-time PCR. Results Transcriptome analysis of 4010 cDNA clones resulted in the identification of the most abundant P. papatasi midgut-specific transcripts. The identified molecules included those with putative roles in digestion and peritrophic matrix formation, among others. Moreover, we identified sandfly midgut transcripts that are expressed only after a blood meal, such as microvilli associated-like protein (PpMVP1, PpMVP2 and PpMVP3, a peritrophin (PpPer1, trypsin 4 (PpTryp4, chymotrypsin PpChym2, and two unknown proteins. Of interest, many of these overabundant transcripts such as PpChym2, PpMVP1, PpMVP2, PpPer1 and PpPer2 were of lower abundance when the sandfly was given a blood meal in the presence of L. major. Conclusion This tissue-specific transcriptome analysis provides a comprehensive look at the repertoire of transcripts present in the midgut of the sandfly P. papatasi. Furthermore, the customized bioinformatic analysis allowed us to compare and identify the overall transcript abundance from sugar-fed, blood-fed and Leishmania-infected sandflies. The suggested upregulation of specific

  13. Cantharidin Impedes Activity of Glutathione S-Transferase in the Midgut of Helicoverpa armigera Hübner

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    Ya Lin Zhang

    2013-03-01

    Full Text Available Previous investigations have implicated glutathione S-transferases (GSTs as one of the major reasons for insecticide resistance. Therefore, effectiveness of new candidate compounds depends on their ability to inhibit GSTs to prevent metabolic detoxification by insects. Cantharidin, a terpenoid compound of insect origin, has been developed as a bio-pesticide in China, and proves highly toxic to a wide range of insects, especially lepidopteran. In the present study, we test cantharidin as a model compound for its toxicity, effects on the mRNA transcription of a model Helicoverpa armigera glutathione S-transferase gene (HaGST and also for its putative inhibitory effect on the catalytic activity of GSTs, both in vivo and in vitro in Helicoverpa armigera, employing molecular and biochemical methods. Bioassay results showed that cantharidin was highly toxic to H. armigera. Real-time qPCR showed down-regulation of the HaGST at the mRNA transcript ranging from 2.5 to 12.5 folds while biochemical assays showed in vivo inhibition of GSTs in midgut and in vitro inhibition of rHaGST. Binding of cantharidin to HaGST was rationalized by homology and molecular docking simulations using a model GST (1PN9 as a template structure. Molecular docking simulations also confirmed accurate docking of the cantharidin molecule to the active site of HaGST impeding its catalytic activity.

  14. Pyrosequencing the Midgut Transcriptome of the Banana Weevil Cosmopolites sordidus (Germar) (Coleoptera: Curculionidae) Reveals Multiple Protease-Like Transcripts.

    Science.gov (United States)

    Valencia, Arnubio; Wang, Haichuan; Soto, Alberto; Aristizabal, Manuel; Arboleda, Jorge W; Eyun, Seong-Il; Noriega, Daniel D; Siegfried, Blair

    2016-01-01

    The banana weevil Cosmopolites sordidus is an important and serious insect pest in most banana and plantain-growing areas of the world. In spite of the economic importance of this insect pest very little genomic and transcriptomic information exists for this species. In the present study, we characterized the midgut transcriptome of C. sordidus using massive 454-pyrosequencing. We generated over 590,000 sequencing reads that assembled into 30,840 contigs with more than 400 bp, representing a significant expansion of existing sequences available for this insect pest. Among them, 16,427 contigs contained one or more GO terms. In addition, 15,263 contigs were assigned an EC number. In-depth transcriptome analysis identified genes potentially involved in insecticide resistance, peritrophic membrane biosynthesis, immunity-related function and defense against pathogens, and Bacillus thuringiensis toxins binding proteins as well as multiple enzymes involved with protein digestion. This transcriptome will provide a valuable resource for understanding larval physiology and for identifying novel target sites and management approaches for this important insect pest.

  15. TAT improves in vitro transportation of fortilin through midgut and into hemocytes of white shrimp Litopenaeus vannamei

    Science.gov (United States)

    Zhou, Yi; Zhang, Wenbing; Mai, Kangsen; Xu, Wei; Zhang, Yanjiao; Ai, Qinghui; Wang, Xiaojie

    2012-06-01

    Fortilin is a multifunctional protein implicated in many important cellular processes. Since injection of Pm-fortilin reduces shrimp mortality caused by white spot syndrome virus (WSSV), there is potential application of fortilin in shrimp culture. In the present study, in order to improve trans-membrane transportation efficiency, the protein transduction domain of the transactivator of transcription (TAT) peptide was fused to fortilin. The Pichia pastoris yeast expression system, which is widely accepted in animal feeds, was used for production of recombinant fusion protein. Green fluorescence protein (GFP) was selected as a reporter because of its intrinsic visible fluorescence. The fortilin, TAT and GFP fusion protein were constructed. Their trans-membrane transportation efficiency and effects on immune response of shrimp were analyzed in vitro. Results showed that TAT peptide improved in vitro uptake of fortilin into the hemocytes and midgut of Litopenaeus vannamei. The phenoloxidase (PO) activity of hemocytes incubated with GFP-Fortilin or GFP-Fortilin-TAT was significantly increased compared with that in the control without expressed fortilin. The PO activity of hemocytes incubated with 200 μg mL-1 GFP-Fortilin-TAT was significantly higher than that in the group with the same concentration of GFP-Fortilin. Hemocytes incubated with GFP-Fortilin-TAT at all concentrations showed significantly higher nitric oxide synthase (NOS) activity than those in the control or in the GFP-Fortilin treatment. The present in vitro study indicated that TAT fusion protein improved the immune effect of fortilin.

  16. A trypsin-like proteinase in the midgut of Ectomyelois ceratoniae Zeller (Lepidoptera: Pyralidae): purification, characterization, and host plant inhibitors.

    Science.gov (United States)

    Ranjbar, Mina; Zibaee, Arash; Sendi, Jalal Jalali

    2014-01-01

    A trypsin-like proteinase was purified and characterized in the midgut of Ectomyelois ceratoniae. A purification process that used Sepharyl G-100 and DEAE-cellulose fast flow chromatographies revealed a proteinase with specific activity of 66.7 μmol/min/mg protein, recovery of 27.04 and purification fold of 23.35. Molecular weight of the purified protein was found to be 35.8 kDa. Optimal pH and temperature were obtained 9 and 20°C for the purified trypsin proteinase, respectively. The purified enzyme was significantly inhibited by PMSF, TLCK, and SBTI as specific inhibitors of trypsins in which TLCK showed the highest inhibitory effect. Trypsin proteinase inhibitors were extracted from four varieties of pomegranate including Brait, Torsh-Sabz, May-Khosh, and Shirin by ion exchange chromatography. It was found that fractions 17-20 of Brait; fractions 18 and 21-26 of Torsh-Sabz; fractions 1-7, 11-17, and 19-21 of May-Khosh and fraction 8 for Shirin showed presence of trypsin inhibitor in these host. Comparison of their inhibitory effects on the purified trypsin proteinase of E. ceratoniae demonstrated that fractions from May-khosh variety had the highest effect on the enzyme among other extracted fractions. Characterization of serine proteinases of insects mainly trypsins is one of the promising methods to decrease population and damages via extracting their inhibitors and providing resistant varieties.

  17. Pyrosequencing the Midgut Transcriptome of the Banana Weevil Cosmopolites sordidus (Germar (Coleoptera: Curculionidae Reveals Multiple Protease-Like Transcripts.

    Directory of Open Access Journals (Sweden)

    Arnubio Valencia

    Full Text Available The banana weevil Cosmopolites sordidus is an important and serious insect pest in most banana and plantain-growing areas of the world. In spite of the economic importance of this insect pest very little genomic and transcriptomic information exists for this species. In the present study, we characterized the midgut transcriptome of C. sordidus using massive 454-pyrosequencing. We generated over 590,000 sequencing reads that assembled into 30,840 contigs with more than 400 bp, representing a significant expansion of existing sequences available for this insect pest. Among them, 16,427 contigs contained one or more GO terms. In addition, 15,263 contigs were assigned an EC number. In-depth transcriptome analysis identified genes potentially involved in insecticide resistance, peritrophic membrane biosynthesis, immunity-related function and defense against pathogens, and Bacillus thuringiensis toxins binding proteins as well as multiple enzymes involved with protein digestion. This transcriptome will provide a valuable resource for understanding larval physiology and for identifying novel target sites and management approaches for this important insect pest.

  18. Analysis of differentially expressed genes between fluoride-sensitive and fluoride-endurable individuals in midgut of silkworm, Bombyx mori.

    Science.gov (United States)

    Qian, Heying; Li, Gang; He, Qingling; Zhang, Huaguang; Xu, Anying

    2016-08-15

    Fluoride tolerance is an economically important trait of silkworm. Near-isogenic lines (NILs) of the dominant endurance to fluoride (Def) gene in Bombyx mori has been constructed before. Here, we analyzed the gene expression profiles of midgut of fluoride-sensitive and fluoride-endurable individuals of Def NILs by using high-throughput Illumina sequencing technology and bioinformatics tools, and identified differentially expressed genes between these individuals. A total of 3,612,399 and 3,567,631 clean tags for the libraries of fluoride-endurable and fluoride-sensitive individuals were obtained, which corresponded to 32,933 and 43,976 distinct clean tags, respectively. Analysis of differentially expressed genes indicates that 241 genes are differentially expressed between the two libraries. Among the 241 genes, 30 are up-regulated and 211 are down-regulated in fluoride-endurable individuals. Pathway enrichment analysis demonstrates that genes related to ribosomes, pancreatic secretion, steroid biosynthesis, glutathione metabolism, steroid biosynthesis, and glycerolipid metabolism are down-regulated in fluoride-endurable individuals. qRT-PCR was conducted to confirm the results of the DGE. The present study analyzed differential expression of related genes and tried to find out whether the crucial genes were related to fluoride detoxification which might elucidate fluoride effect and provide a new way in the fluorosis research.

  19. Brush border membrane binding properties of Bacillus thuringiensis Vip3A toxin to Heliothis virescens and Helicoverpa zea midguts.

    Science.gov (United States)

    Lee, Mi Kyong; Miles, Paul; Chen, Jeng-Shong

    2006-01-27

    The binding properties of Vip3A, a new family of Bacillus thuringiensis insecticidal toxins, have been examined in the major cotton pests, Heliothis virescens and Helicoverpa zea. Vip3A bound specifically to brush border membrane vesicles (BBMV) prepared from both insect larval midguts. In order to examine the cross-resistance potential of Vip3A to the commercially available Cry1Ac and Cry2Ab2 toxins, the membrane binding site relationship among these toxins was investigated. Competition binding assays demonstrated that Vip3A does not inhibit the binding of either Cry1Ac or Cry2Ab2 and vice versa. BBMV protein blotting experiments showed that Vip3A does not bind to the known Cry1Ac receptors. These distinct binding properties and the unique protein sequence of Vip3A support its use as a novel insecticidal agent. This study indicates a very low cross-resistance potential between Vip3A and currently deployed Cry toxins and hence supports its use in an effective resistance management strategy in cotton.

  20. Specific binding of Bacillus thuringiensis Cry2A insecticidal proteins to a common site in the midgut of Helicoverpa species.

    Science.gov (United States)

    Hernández-Rodríguez, Carmen Sara; Van Vliet, Adri; Bautsoens, Nadine; Van Rie, Jeroen; Ferré, Juan

    2008-12-01

    For a long time, it has been assumed that the mode of action of Cry2A toxins was unique and different from that of other three-domain Cry toxins due to their apparent nonspecific and unsaturable binding to an unlimited number of receptors. However, based on the homology of the tertiary structure among three-domain Cry toxins, similar modes of action for all of them are expected. To confirm this hypothesis, binding assays were carried out with (125)I-labeled Cry2Ab. Saturation assays showed that Cry2Ab binds in a specific and saturable manner to brush border membrane vesicles (BBMVs) of Helicoverpa armigera. Homologous-competition assays with (125)I-Cry2Ab demonstrated that this toxin binds with high affinity to binding sites in H. armigera and Helicoverpa zea midgut. Heterologous-competition assays showed a common binding site for three toxins belonging to the Cry2A family (Cry2Aa, Cry2Ab, and Cry2Ae), which is not shared by Cry1Ac. Estimation of K(d) (dissociation constant) values revealed that Cry2Ab had around 35-fold less affinity than Cry1Ac for BBMV binding sites in both insect species. Only minor differences were found regarding R(t) (concentration of binding sites) values. This study questions previous interpretations from other authors performing binding assays with Cry2A toxins and establishes the basis for the mode of action of Cry2A toxins.

  1. Specific Binding of Bacillus thuringiensis Cry2A Insecticidal Proteins to a Common Site in the Midgut of Helicoverpa Species▿

    Science.gov (United States)

    Hernández-Rodríguez, Carmen Sara; Van Vliet, Adri; Bautsoens, Nadine; Van Rie, Jeroen; Ferré, Juan

    2008-01-01

    For a long time, it has been assumed that the mode of action of Cry2A toxins was unique and different from that of other three-domain Cry toxins due to their apparent nonspecific and unsaturable binding to an unlimited number of receptors. However, based on the homology of the tertiary structure among three-domain Cry toxins, similar modes of action for all of them are expected. To confirm this hypothesis, binding assays were carried out with 125I-labeled Cry2Ab. Saturation assays showed that Cry2Ab binds in a specific and saturable manner to brush border membrane vesicles (BBMVs) of Helicoverpa armigera. Homologous-competition assays with 125I-Cry2Ab demonstrated that this toxin binds with high affinity to binding sites in H. armigera and Helicoverpa zea midgut. Heterologous-competition assays showed a common binding site for three toxins belonging to the Cry2A family (Cry2Aa, Cry2Ab, and Cry2Ae), which is not shared by Cry1Ac. Estimation of Kd (dissociation constant) values revealed that Cry2Ab had around 35-fold less affinity than Cry1Ac for BBMV binding sites in both insect species. Only minor differences were found regarding Rt (concentration of binding sites) values. This study questions previous interpretations from other authors performing binding assays with Cry2A toxins and establishes the basis for the mode of action of Cry2A toxins. PMID:18931285

  2. Purification, characterization and sequencing of the major beta-1,3-glucanase from the midgut of Tenebrio molitor larvae.

    Science.gov (United States)

    Genta, Fernando A; Bragatto, Ivan; Terra, Walter R; Ferreira, Clélia

    2009-12-01

    The major beta-1,3-glucanase from Tenebrio molitor (TLam) was purified to homogeneity (yield, 6%; enrichment, 113 fold; specific activity, 4.4 U/mg). TLam has a molecular weight of 50 kDa and a pH optimum of 6. It is an endoglucanase that hydrolyzes beta-1,3-glucans as laminarin and yeast beta-1,3-1,6-glucan, but is inactive toward other polysaccharides (as unbranched beta-1,3-glucans or mixed beta-1,3-1,4-glucan from cereals) or disaccharides. The enzyme is not inhibited by high substrate concentrations and has low processivity (0.6). TLam has two ionizable groups involved in catalysis, and His, Tyr and Arg residues plus a divalent ion at the active site. A Cys residue important for TLam activity is exposed after laminarin binding. The cDNA coding for this enzyme was cloned and sequenced. It belongs to glycoside hydrolase family 16, and is related to other insect glucanases and glucan-binding proteins. Sequence analysis and homology modeling allowed the identification of some residues (E174, E179, H204, Y304, R127 and R181) at the active site of the enzyme, which may be important for TLam activity. TLam efficiently lyses fungal cells, suggesting a role in making available walls and cell contents to digestion and in protecting the midgut from pathogen infections.

  3. Microflora de Bromeliáceas do estado de Pernambuco, Brasil

    Directory of Open Access Journals (Sweden)

    Luis Tavares de Lyra

    1976-01-01

    Full Text Available O autor estudou a microflora de dois gêneros de BROMELIACEAE: Hoenbergia e Portea. As coletas do material foram feitas em seis regiões do Estado de Pernambuco; 1 Região da Mata-Úmida; 2 Região da Mata-Seca; 3 Região do Agreste Central; 4 Região do Agreste Setentrional; 5 Região do Agreste Meridional; 6 Região do Recife. As seguintes diatomáceas indicadoras de águas poluídas (espécies oligossaprobias foram encontradas nas seis regiões estudadas: Gomphonema parvulum (Kutz Grunow., Hantzschia amphioxys Grunow, Pinnularia borealis Ehr., Pinnularia microstauron (Ehr Cleve, gomphonema gracile Ehr., Nitzschia palea Kutz., Melosira roeseana Rabenh., Navicula mutica Kutz., Navicula cryptocephala Kutz., Eunotia pectinalis (Kutz Rabenh. Foram também observadas CHLOROPHYCEAS nas estações chuvosa e seca nas diversas regiões. Algumas são indicadoras de oligossaprobidade: Scenedesmus quadricauda (Turpin brebisson. Chlorococcum sp., Chlorella sp. Os fatores ecológicos e comentários referentes ás diatomáceas foram anotados no texto. A tabela I indica a frequência das diatomáceas nas seis regiões estudadas. Maior número dessas diatomáceas, registramos nas regiões do Agreste. A tabela II mostra a temperatura e pH da água de Hoenbergia e Portea em ambas as estações do ano (inverno e verão. Observamos a ocorrência de larvas de culex em Portea e hoenbergia, entretanto, raramente encontramos larvas de Anopheles. As coletas foram feitas durante as estações chuvosa e seca em Hoenbergia e Portea. Determinamos 35 espécies provenientes de 78 amostras coletadas durante o período de 26 meses.The author studied the microflora from two genera of Bromeliaceae: Hoenbergia and Portea. The material was collected in six natural regions of Pernambuco State: 1 Humid Forest Region; 2 Dry forest region; 3 Central Agreste Region; 4 Setentrional Agreste Region; 5 Meridional Agreste Region; 6 Recife Region. The following diatoms that indicate polluted

  4. Bifidogenic effect of grain larvae extract on serum lipid, glucose and intestinal microflora in rats

    Indian Academy of Sciences (India)

    Sang-Oh Park; Byung-Sung Park

    2015-09-01

    The main objective of this study was to investigate whether orally administered Korean grain larvae ethanol extract (GLE) had a bifidogenic effect in normal rats. Male Sprague–Dawley rats were divided into a negative control group (CO) and GLE orally administered (5.0, 7.0 and 9.0 mg/100 g body weight) groups. Thymus and spleen weights dosedependently increased by 128.58% and 128.58%, respectively, but abdominal fat decreased by 19.18% after GLE administration compared with that in the CO group (<0.05). Serum triglycerides, total cholesterol, low-density lipoprotein cholesterol, and glucose decreased by 30.26%, 7.33%, 27.20%, and 6.96%, respectively, whereas highdensity lipoprotein cholesterol increased by 129.93% in the GLE groups compared with those in the CO group (<0.05). IgG, IgM, IgA in the GLE groups increased 203.68%, 181.41%, and 238.25%, respectively, compared to that in the CO group (<0.05). Bifidobacteria and Lactobacillus increased by 115.74% and 144.28%, whereas Bacteroides, Clostridium, Escherichia, and Streptococcus decreased by 17.37%, 17.46%, 21.25%, and 19.16%, respectively, in the GLE groups compared with those in the CO group (<0.05). Total organic acids, acetic acid, and propionic acid increased by 151.40%, 188.09%, and 150.17%, whereas butyric acid and valeric acid decreased by 40.65% and 49.24%, respectively, in the GLE groups as compared with those in the CO group (<0.05). These results suggest that Korean GLE improves the bifidogenic effect by increasing cecal organic acids and modulating gut microflora via a selective increase in Bifidobacterium in normal rats.

  5. Feasibility study on fermentative conversion of raw and hydrolyzed starch to hydrogen using anaerobic mixed microflora

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Ching-Hsiung [Department of Biological Engineering, Yung Ta Institute of Technology and Commerce, Pingtung (China); Lu, Wei-Bin [Department of Cosmetic Science, Chung Hwa College of Medical Technology, Tainan (China); Chang, Jo-Shu [Department of Chemical Engineering, National Cheng Kung University, Tainan 701 (China)

    2007-11-15

    In this work, H{sub 2} was produced by anaerobic mixed microflora with phosphate-buffered medium containing starch or enzyme-treated starch hydrolyzate as the carbon substrate. The effect of pH on H{sub 2}-producing performance was examined for cultures converting raw starch or hydrolyzed starch into H{sub 2}. Response surface methodology was utilized to determine the best condition (41 C, pH 5.2, 2.1% (v/v) enzyme dosage, 27 h reaction time) for starch hydrolysis with concentrated crude amylase obtained from Bacillus subtilis ATCC 21332. The mixed culture was able to produce H{sub 2} at an optimal pH of 7.0 irrespective of raw or hydrolyzed starch. Direct starch fermentation attained a highest maximum H{sub 2} production rate (R{sub max}), overall H{sub 2} production rate (R{sub overall}), and H{sub 2} yield (Y{sub H2}) of 25.6 ml/h, 88 ml/h/l, and 5.28 mmol H{sub 2}/g starch (4.64 mmol H{sub 2}/g COD), respectively. In contrast, using hydrolyzed starch as the substrate gave rise to much better H{sub 2} producing performance, as the highest R{sub max}, R{sub overall}, and Y{sub H2} values increased to 43.1 ml/h, 210 ml/h/l, and 6.1 mmol H{sub 2}/g COD, respectively. This clearly demonstrates the advantage of using hydrolyzed starch for fermentative H{sub 2} production. The soluble metabolites consisted primarily of acetate (HAc), ethanol (EtOH), butyrate (HBu), and 2,3 butandiol. The amount of H{sub 2} produced from raw and hydrolyzed starch (especially, raw starch) could be estimated from formation of HAc and HBu known to stoichiometrically correlate with H{sub 2} production. (author)

  6. [The bacterial microflora of diabetic foot infection and factors determining its spectrum in Ouagadougou (Burkina Faso)].

    Science.gov (United States)

    Guira, O; Tiéno, H; Traoré, S; Diallo, I; Ouangré, E; Sagna, Y; Zabsonré, J; Yanogo, D; Traoré, S S; Drabo, Y J

    2015-12-01

    The aim of the study was to describe the bacterial microflora of diabetic foot infection and to identify the factors which determine the bacterial spectrum in order to increase empiric antibiotic prescription in Ouagadougou. The study was a cross-sectional one, carried from July 1st, 2011 to June 30, 2012 in the departments of internal medicine and general and digestive surgery in Yalgado Ouédraogo teaching hospital. Samples for bacteriological tests consisted of aspiration of pus through the healthy skin, curettage and swab of the base of the ulceration or tissue biopsy from foot lesions. The bacteria's sensitivity to antibiotics has been tested by the qualitative method (Kirby-Bauer). The frequency of diabetic foot infection was 14.45% and the monthly incidence 5.33. The mean age of patients was 56 years and the sex ratio 1.37. Foot ulcerations were chronic in 33 (51.56%), necrotic in 51 (79.69%) and associated with osteitis in 40 (62.5%) patients. Infection was grade 3 in 70.3% cases. Thirty-nine patients had received antibiotics before hospital admission. Among the 71 samples, 62 (87.32%) cultures were positive: 53 (85.48%) monomicrobial and 9 (14.52%) bimicrobial. Aerobic Gram-positive cocci (76%) were the most frequent from ulcerations: Staphylococcus aureus (32.39%), Streptococcus sp (18.30%). Negative coagulase staphylococci have been found in 23.94% cases. Aerobic gram-negative bacilli have been isolated from 24% ulcerations. No factor was associated with the type of bacteria. Gram-positive pathogen cocci showed a high sensitivity to amoxicillin-clavulanic acid and oxacillin. No methicillin-resistant Staphylococcus aureus (MRSA) or extended-spectrum beta lactamase Enterobacteriaceae (ESBL) have been isolated. A better design is necessary to a clarification of bacterial flora in diabetic foot infections. Prevention of bacterial resistance is also needed.

  7. The Effect of Iranian Customary Used Probiotic Yogurt on the Children’s Salivary Cariogenic Microflora

    Directory of Open Access Journals (Sweden)

    Ali Nozari

    2015-06-01

    Full Text Available Statement of the Problem: Dental caries is the most common disease of child-hood. Using probiotics has recently been introduced to reduce the incidence of dental caries.It consists of live microbial food supplements that beneficially affect the host, and hence are considered an alternative way to eradicate the infections. Purpose: The aim of this study is to evaluate the effect of consumption of probi-otic yogurt on the children’s salivary cariogenic microflora. Materials and Method: A double-blind randomized study was performed recruit-ing 2 parallel groups; 24 healthy children in the case and 25 children in the control group. All healthy children were followed- up over 4 periods. Periods 1 and 3 were wash-out periods with duration of 1 and 2 weeks, respectively. During periods 2 and 4 (2weeks duration each, the case group consumed 200g yogurt containing Bifidobacterium lactis (1×106 per gram once daily and the control group consumed normal yogurt. Salivary Streptococci mutans and Lactobacilli were enumerated before and after the yogurt consumption periods. Pre- and post-treatment values within and between regimens were compared using the t-test and paired samples. Results: There was a reduction in Streptococcus mutans and Lactobacillus counts in the control group, but for Streptococcus mutans, the count reduction between phases 1 and 4 was statistically significant (p= 0.009. In the case group, neither the Streptococcus mutans count nor the Lactobacilli count was significantly reduced. Conclusion: Based on the findings of this study, short-term daily consumption of probiotic yogurt containing Bifidobacterium lactis could not reduce the levels of salivary Streptococcus mutans and Lactobacilli in 6 to 12 year-old children, while normal yogurt could reduce the Streptococcus mutans counts significantly.

  8. Effect of dietary honey on intestinal microflora and toxicity of mycotoxins in mice

    Directory of Open Access Journals (Sweden)

    Hegazy Eman M

    2006-03-01

    Full Text Available Abstract Background Bee honey is a functional food which has a unique composition, antimicrobial properties and bifidogenic effect. In order to assess whether honey can inhibit the toxic effect of mycotoxins, the present study was undertaken. Methods Production of biomass and toxins by Aspergillus parasiticus and Aspergillus ochraceus were followed in media without and with honey. Although aflatoxins and ochratoxin A. were administrated to male Swiss albino mice up to 1 μg and 10 ng/kg body weight/day respectively. The experimental animals were fed diets without our with 10% honey for two months. The changes in colonic probiotic bacteria, determintal colon enzyme glucuronidases, and genotoxicity were followed. Results Addition of 32% in its media increased the biomass of A parasiticus, while the biomass of A. ochraceus decreased and Ochratoxin A. was not produced. When the honey was added at the ratio of 32 and 48% in the medium. No relationship was found between mycelium weight and production of mycotoxins. Oral administration of aflatoxins (mixture of B1, B2, G1 and G2 and Ochratoxin A. induced structural and numerical chromosomal aberrations in bone marrow and germ cells of male mice, whereas, honey treatment reduced the genotoxicity of mycotoxins. Also both toxins induced histopathological changes in liver and kidney. Feeding on diet supplemented with honey improved the histopathological changes in case of aflatoxin group, but not in the case of ochratoxin A. group (except of kidney in two cases. No significant differences were found in the activity of colon β-glucuronidase between group fed diet with or without honey. On the other hand, the colon bifido bacteria and lactobacilli counts were increased markedly in group receiving diet supplemented with honey. Conclusion Substituting sugars with honey in processed food can inhibit the harmful and genotoxic effects of mycotoxins, and improve the gut microflora.

  9. Effects of various pretreatment methods on mixed microflora to enhance biohydrogen production from corn stover hydrolysate

    Institute of Scientific and Technical Information of China (English)

    Kun Zhang; Nanqi Ren; Changhong Guo; Aijie Wang; Guangli Cao

    2011-01-01

    Five individual pretreatment methods,including three widely-used protocols (heat,acid and base) and two novel attempts (ultrasonic and ultraviolet),were conducted in batch tests to compare their effects on mixed microflora to enhance hydrogen (H2) production from corn stover hydrolysate.Experimental results indicated that heat and base pretreatments significantly increased H2 yield with the values of 5.03 and 4.45 mmol H2/g sugar utifized,respectively,followed by acid pretreatment of 3.21 mmol H2/g sugar utilized.However,compared with the control (2.70 mmol H2/g sugar utilized),ultrasonic and ultraviolet pretreatments caused indistinctive effects on H2 production with the values of 2.92 and 2.87 mmol H2/g sugar utilized,respectively.The changes of soluble metabolites composition caused by pretreatment were in accordance with H2-producing behavior.Concretely,more acetate accumulation and less ethanol production were found in pretreated processes,meaning that more reduced nicotinamide adenine dinucleotide (NADH) might be saved and flowed into H2-producing pathways.PCR-DGGE analysis indicated that the pretreatment led to the enrichment of some species,which appeared in large amounts and even dominated the microbial community.Most of the dominated species were affiliated to Enterobacter spp.and Escherichia spp.As another efficient H2 producer,Clostridium bifermentan was only found in a large quantity after heat pretreatment.This strain might be mainly responsible for better performance of H2 production in this case.

  10. Effect of microflora and lactose on the absorption of calcium, phosphorus and magnesium in the hindgut of the rat.

    Science.gov (United States)

    Andrieux, C; Sacquet, E

    1983-01-01

    For 4 weeks, 3-month old germfree (GF) and conventional (CV) rats were given a semi-synthetic diet sterilized by irradiation with or without 10% of lactose. During the 5th week, 0.2% of titanium oxide (TiO2) was added to the diet and the rats were killed at regular intervals throughout the light/dark cycle. The patterns of TiO2 and 45Ca excretion were similar, indicating that TiO2 was a good marker of unabsorbed calcium transit. The apparent absorption coefficient of calcium, magnesium and phosphorus was determined in the ileum, caecum, large intestine and faeces by the mineral/TiO2 ratio. The effects of microflora and lactose varied with the mineral and the digestive tract level studied. --In the small intestine, microflora had no effect on the apparent absorption of calcium and magnesium but did have an unfavorable influence on phosphorus absorption. Lactose increased calcium and magnesium absorption, and this increase was similar in GF and CV rats, but lactose had a favorable effect on phosphorus absorption only in CV rats. --In the caecum, microflora had an unfavorable effect on the apparent absorption of calcium and magnesium and a favorable effect on phosphorus absorption. The ingestion of lactose reduced calcium and magnesium absorption in the caecum of GF rats and phosphorus absorption in the caecum of CV animals. --In the colon, mineral absorption was not significant in either CV or GF rats receiving the lactose-free diets. Lactose ingestion caused the absorption of calcium, magnesium and phosphorus to rise significantly only in GF rats. This absorption contributed to the stronger effect of lactose on total calcium and phosphorus absorption in GF rats.

  11. The Effects of Hormones and Vaginal Microflora on the Glycome of the Female Genital Tract: Cervical-Vaginal Fluid

    Science.gov (United States)

    Moncla, Bernard J.; Chappell, Catherine A.; Debo, Brian M.; Meyn, Leslie A.

    2016-01-01

    In this study, we characterized the glycome of cervical-vaginal fluid, collected with a Catamenial cup. We quantified: glycosidase levels; sialic acid and high mannose specific lectin binding; mucins, MUC1, MUC4, MUC5AC, MUC7; and albumin in the samples collected. These data were analyzed in the context of hormonal status (day of menstrual cycle, hormonal contraception use) and role, if any, of the type of the vaginal microflora present. When the Nugent score was used to stratify the subjects by microflora as normal, intermediate, or bacterial vaginosis, several important differences were observed. The activities of four of six glycosidases in the samples from women with bacterial vaginosis were significantly increased when compared to normal or intermediate women: sialidase, P = IUDs (P = 0.02). The number of sialic acid binding sites was lower in the postmenopausal group (P = 0.04), but the number of high mannose binding sites, measured with Griffithsin, was not significantly different among the 6 hormonal groups. The glycosidase levels in the cervical-vaginal mucus were rather low in the groups, with exception of α-glucosidase activity that was much lower in the postmenopausal group (Pmicroorganisms. These effects were so influential that it required us to remove subjects with BV for data interpretation of the impact of hormones. We also suggest that certain changes occurring in vaginal/cervical proteins are due to bacteria or their products. Therefore, the quantitation of vaginal mucins and lectin binding offers a new method to monitor bacteria-host interactions in the female reproductive tract. The data suggest that some of the changes in these components are the result of host processing, such as the increases in mucin content, while the microflora is responsible for the increases in glycosidases and the decreases in lectin binding. The methods should be considered a valid marker for insult to the female genital tract. PMID:27437931

  12. Effect of red pepper Capsicum annuum var. conoides and garlic Allium sativum on plasma lipid levels and cecal microflora in mice fed beef tallow.

    Science.gov (United States)

    Kuda, Takashi; Iwai, Akiko; Yano, Toshihiro

    2004-10-01

    Antihyperlipidemia or hypocholesterolaemic and antibacterial activities of red hot pepper and garlic are well known. To determine the effect of the dietary spices ingested to suppress blood lipids on the intestinal condition, we examined plasma lipid levels and cecal microflora in mice that were fed diets containing 19% (w/w) beef tallow and 2% red pepper Capsicum annuum var. conoides 'Takanotume' (RP) or garlic Allium sativum 'White' (GP) for 4-weeks. Plasma triacylglyceride level was suppressed by the spices. RP lowered cecal bacteroidaceae, a predominant bacterial group (from 9.4 to 9.0 log CFU/g), bifidobacteria (from 8.7 to 7.6 log CFU/g), and staphylococci. Although GP increased the cecal weight including their contents, significant differences were not shown in the cecal microflora. These results suggest that RP can affect the intestinal condition and host health through the disturbance of intestinal microflora.

  13. Influence of microflora on texture and contents of amino acids, organic acids, and volatiles in semi-hard cheese made with DL-starter and propionibacteria

    DEFF Research Database (Denmark)

    Rehn, Lina Ulrika Ingeborg; Vogensen, Finn Kvist; Persson, S.-E.;

    2011-01-01

    for the cheese variety Grevé was obtained by using a PAB culture in combination with different DL-starters and making the cheeses at 2 dairy plants with different time and temperature profiles during ripening. Propionic acid bacteria dominated the microflora during ripening after a warm room period at levels......The microflora of semi-hard cheese made with DL-starter and propionic acid bacteria (PAB) is quite complex, and we investigated the influence of its variation on texture and contents of organic acids, free amino acids, and volatile compounds. Variation in the microflora within the normal range......, propionate, total content of free amino acids, 2-propanol, and ethyl propionate in the ripened cheeses were related to the number of PAB. A decrease in the relative content of Asp and Lys and increase of Phe over the ripening time were different from what is observed in semi-hard cheese without PAB...

  14. Potential DMSP-degrading Roseobacter clade dominates endosymbiotic microflora of Pyrodinium bahamense var. compressum (Dinophyceae) in vitro.

    Science.gov (United States)

    Onda, Deo Florence L; Azanza, Rhodora V; Lluisma, Arturo O

    2015-09-01

    Many aspects of the biology and ecology of the toxic dinoflagellate Pyrodinium bahamense var. compressum are still poorly understood. In this brief note, we present identification of its associated intracellular bacteria or endosymbionts via PCR cloning and 16s rRNA gene sequencing and their localization by confocal microscopy, a first for Pyrodinium. The most frequently observed species in the endosymbiotic microflora were from Roseobacter clade (Alphaproteobacteria, 68%) and Gilvibacter sediminis (Flavobacteriaceae, 20%). Roseobacter lineage, the most abundant taxa in this study, is known to be involved in dimethylsulfoniopropionate metabolism which is highly produced in dinoflagellates-a possible strong factor shaping the structure of the associated bacterial community.

  15. Effect of antimicrobial spice and herb extract combinations on Listeria monocytogenes, Staphylococcus aureus, and spoilage microflora growth on cooked ready-to-eat vacuum-packaged shrimp.

    Science.gov (United States)

    Weerakkody, Nimsha S; Caffin, Nola; Dykes, Gary A; Turner, Mark S

    2011-07-01

    Two spice and herb extract combinations from galangal (Alpinia galanga), rosemary (Rosmarinus officinalis), and lemon iron bark (Eucalyptus staigerana) were evaluated for their ability to inhibit the growth of inoculated Listeria monocytogenes and Staphylococcus aureus and naturally present spoilage microflora on cooked ready-to-eat shrimp stored for 16 days at 4 or 8 °C. A combination of galangal, rosemary, and lemon iron bark significantly reduced (P galangal and rosemary extract significantly reduced (P galangal, rosemary, and lemon iron bark extracts can be used to control the growth of spoilage microflora on ready-to-eat shrimp.

  16. Methanogens Consortium Analysis in Low-temperature Biogas Functional Microflora%低温沼气功能菌群的产甲烷菌分析

    Institute of Scientific and Technical Information of China (English)

    丁建南; 田晓娟; 张国华; 黄黄; 黄江丽

    2012-01-01

    运用16s rDNA PCR和实时荧光qPCR技术,对选育的低温沼气功能菌群中的产甲烷菌种群结构和数量进行了研究,并与常温沼气功能菌群进行了比较分析。在总共33个产甲烷菌分类单元中低温沼气功能菌群占有20个,低温沼气功能菌群的产甲烷菌多样性远高于常温沼气功能菌群。低温沼气功能菌群以对温度要求较低的中低温产甲烷菌为主,而常温沼气功能菌群则以中高温产甲烷菌占优。虽然两类菌群的产甲烷菌在多样性和种群结构方面存在着较大差异,但在产甲烷菌数量方面并没有明显的不同,这表明选育的低温沼气功能菌群和常温沼气功能菌群一样,已经形成了稳定的产甲烷菌种群。%Methanogens populations in low and normal temperature biogas functional microfloras were respectively investigated using 16s rDNA PCR and real-time qPCR techniques.The 16s rDNA clone library of methanogens showed a great difference in methanogens consortium diversity and structure between the two microfloras.A total of 33 OUTs of methanogens were identified,including 20 in low-temperature microflora and 13 in normal temperature microflora.Methanogens requiring low-medium temperature were found prevalent in low-temperature microflora and those active at moderate-high temperature occurred mainly in normal temperature microflora.However,the results of real-time qPCR showed that there was no significant difference in methanogen quantity between the two studied microfloras,indicating a stable methanogenic community had been built in both microfloras.

  17. Comparative proteomic analysis of Aedes aegypti larval midgut after intoxication with Cry11Aa toxin from Bacillus thuringiensis.

    Directory of Open Access Journals (Sweden)

    Angeles Cancino-Rodezno

    Full Text Available Cry toxins produced by Bacillus thuringiensis bacteria are environmentally safe alternatives to control insect pests. They are pore-forming toxins that specifically affect cell permeability and cellular integrity of insect-midgut cells. In this work we analyzed the defensive response of Aedes aegypti larva to Cry11Aa toxin intoxication by proteomic and functional genomic analyses. Two dimensional differential in-gel electrophoresis (2D-DIGE was utilized to analyze proteomic differences among A. aegypti larvae intoxicated with different doses of Cry11Aa toxin compared to a buffer treatment. Spots with significant differential expression (p<0.05 were then identified by liquid chromatography-tandem mass spectrometry (LC-MS/MS, revealing 18 up-regulated and seven down-regulated proteins. The most abundant subcategories of differentially expressed proteins were proteins involved in protein turnover and folding, energy production, and cytoskeleton maintenance. We selected three candidate proteins based on their differential expression as representatives of the different functional categories to perform gene silencing by RNA interference and analyze their functional role. The heat shock protein HSP90 was selected from the proteins involved in protein turnover and chaperones; actin, was selected as representative of the cytoskeleton protein group, and ATP synthase subunit beta was selected from the group of proteins involved in energy production. When we affected the expression of ATP synthase subunit beta and actin by silencing with RNAi the larvae became hypersensitive to toxin action. In addition, we found that mosquito larvae displayed a resistant phenotype when the heat shock protein was silenced. These results provide insight into the molecular components influencing the defense to Cry toxin intoxication and facilitate further studies on the roles of identified genes.

  18. Generation of a Transcriptome in a Model Lepidopteran Pest, Heliothis virescens, Using Multiple Sequencing Strategies for Profiling Midgut Gene Expression.

    Directory of Open Access Journals (Sweden)

    Omaththage P Perera

    Full Text Available Heliothine pests such as the tobacco budworm, Heliothis virescens (F., pose a significant threat to production of a variety of crops and ornamental plants and are models for developmental and physiological studies. The efforts to develop new control measures for H. virescens, as well as its use as a relevant biological model, are hampered by a lack of molecular resources. The present work demonstrates the utility of next-generation sequencing technologies for rapid molecular resource generation from this species for which lacks a sequenced genome. In order to amass a de novo transcriptome for this moth, transcript sequences generated from Illumina, Roche 454, and Sanger sequencing platforms were merged into a single de novo transcriptome assembly. This pooling strategy allowed a thorough sampling of transcripts produced under diverse environmental conditions, developmental stages, tissues, and infections with entomopathogens used for biological control, to provide the most complete transcriptome to date for this species. Over 138 million reads from the three platforms were assembled into the final set of 63,648 contigs. Of these, 29,978 had significant BLAST scores indicating orthologous relationships to transcripts of other insect species, with the top-hit species being the monarch butterfly (Danaus plexippus and silkworm (Bombyx mori. Among identified H. virescens orthologs were immune effectors, signal transduction pathways, olfactory receptors, hormone biosynthetic pathways, peptide hormones and their receptors, digestive enzymes, and insecticide resistance enzymes. As an example, we demonstrate the utility of this transcriptomic resource to study gene expression profiling of larval midguts and detect transcripts of putative Bacillus thuringiensis (Bt Cry toxin receptors. The substantial molecular resources described in this study will facilitate development of H. virescens as a relevant biological model for functional genomics and for new

  19. Influence of zinc nanoparticles on survival of worms Eisenia fetida and taxonomic diversity of the gut microflora.

    Science.gov (United States)

    Yausheva, Еlena; Sizova, Еlena; Lebedev, Svyatoslav; Skalny, Anatoliy; Miroshnikov, Sergey; Plotnikov, Andrey; Khlopko, Yuri; Gogoleva, Natalia; Cherkasov, Sergey

    2016-07-01

    The study was conducted to examine the effect of zinc nanoparticles on survival of worms Eisenia fetida and composition of the gut microflora. Analysis of the survival data has shown that the introduction of high doses of the nanoparticles causes death of worms in the second group with 35 % mortality rate and activates protective mechanisms realized as mucous film. DNA from the worm guts was extracted and 16S metagenomic sequencing was fulfilled using MiSeq (Illumina). Regarding the gut microflora of worms in the control group, high diversity of microorganisms (303 OTUs) was noted. Most of those belong to the taxa Firmicutes (51.9 % of the total high-quality united reads), Proteobacteria (24.1 % of the total), and Actinobacteria (13.3 % of the total), which were represented by numerous species of gen. Clostridium (C. saccharobutylicum, C. saccharoperbutylacetonicum, C. beijerinckii), gen. Pseudomonas (P. hydrogenovora, P. aeruginosa, and P. putida), gen. Bacillus (B. megaterium, B. silvestris), gen. Cellulomonas (B. megaterium, B. silvestris), and other numerically smaller genera. Adding of zinc nanoparticles to the substrate decreased the diversity of bacteria (78 OTUs) as well as percentage of bacteria belonging to the taxon Firmicutes (-41.6 %) and increased the proportion of Proteobacteria due to growth in abundance of gen. Verminephrobacter (+46 %) and gen. Ochrobactrum (+19.5 %).

  20. Effects of dietary supplementation with Gynura procumbens (Merr.) on egg yolk cholesterol, excreta microflora and laying hen performance.

    Science.gov (United States)

    Lokhande, A; Ingale, S L; Lee, S H; Sen, S; Khong, C; Chae, B J; Kwon, I K

    2014-01-01

    Abstract 1. The present study investigated the effects of dietary supplementation with Gynura procumbens on egg yolk and serum cholesterol and triglycerides, excreta microflora, laying performance and egg quality. 2. A total of 160 Hy-Line Brown layers (45 weeks old) were randomly assigned into 4 treatments on the basis of laying performance. Each treatment had 4 replicates with 10 birds each. 3. Dietary treatments were basal diet supplemented with 0 (control), 2.5, 5.0 and 7.5 g/kg diet G. procumbens during 56-d feeding period. 4. Serum (d 21, 42 and 56) and egg yolk (d 28, 42 and 56) cholesterol and triglycerides concentrations were linearly reduced with increasing dietary concentrations of G. procumbens. 5. Increasing dietary concentrations of G. procumbens linearly reduced the excreta total anaerobic bacteria (d 28), Clostridium sp. and Escherichia coli (d 28 and 56) populations. 6. Overall egg production and egg mass were linearly increased, and overall feed efficiency was linearly improved with increase in dietary G. procumbens. 7. Dietary increasing concentrations of G. procumbens linearly improved egg yolk colour (d 28 and 56) and breaking strength of eggs (d 56). 8. The results obtained in the present experiment indicate that dietary supplementation with G. procumbens could reduce the egg yolk cholesterol, suppresses harmful excreta microflora and improves layers performance.

  1. Comparative evaluation of subgingival plaque microflora in pregnant and non-pregnant women: A clinical and microbiologic study

    Directory of Open Access Journals (Sweden)

    Rishi Emmatty

    2013-01-01

    Full Text Available Context: Gingival changes in pregnancy have been attributed to changes in the subgingival biofilm related to hormonal variations. Aims: To evaluate the subgingival plaque microflora in pregnant and nonpregnant women to determine if pregnancy induces any alterations in the subgingival plaque and to associate these changes with changes in periodontal status. Settings and Design: Thirty pregnant and 10 nonpregnant women within the age group of 20-35 years having a probing pocket depth (PPD of 3-4 mm were included in the study. The pregnant women were equally divided into 3 groups of 10, each belonging to I, II, and III trimester. Materials and Methods: Plaque index, gingival index, PPD, and microbiologic evaluation for specific bacterial counts for Prevotella intermedia, Porphyromonas gingivalis, Aggregatibacter actinomycetemcomitans, Parvimonas micra, and Fusobacterium nucleatum were carried out for all subjects. Statistical Analysis: Mann-Whitney U test. Results: Increase in gingival inflammation was observed in II and III trimester as compared with I trimester and control. Plaque scores did not show any significant difference between pregnant and nonpregnant women. Specific bacterial examination revealed an increase in proportion of P. intermedia in pregnant women of both II and III trimester as compared with I trimester and nonpregnant women. Conclusions: A definite increase in proportions of P. intermedia occurs in subgingival plaque microflora in pregnancy that may be responsible for the exaggerated gingival response.

  2. Intestinal microflora and allergic diseases in children%肠道菌群与儿童过敏性疾病

    Institute of Scientific and Technical Information of China (English)

    王超颖

    2015-01-01

    肠道菌群在调节机体免疫功能、维持肠道黏膜屏障、抑制外源性致病菌定植等方面具有重要作用.近年来流行病学调查和实验研究提示生命早期肠道菌群的紊乱与过敏性疾病的发生发展有密切关系.关于益生菌对过敏性疾病的预防和治疗作用,近年来已有多项研究,但尚无确切结论.%Intestinal microflora play a significant role in regulating systemic immune function,maintaining the intestinal barrier,and preventing the invasion of external pathogenic bacteria.Recent epidemiological studies and experimental research suggest that the occurrence and development of allergic diseases is associated with the disturbances in the intestinal microflora during early life.Research has been taken to evaluate the prevention and treatment effects in allergic disease of probiotics,which is currently unestablished.

  3. Breeding and Testing of Hypothermic Biogas Functional Microflora%低温沼气功能菌群的选育与试验

    Institute of Scientific and Technical Information of China (English)

    丁建南; 于一尊; 黄江丽; 田晓娟; 张国华; 张志红

    2012-01-01

    Two groups of hypothermic biogas functional microflora were bred from anaerobic environmental and digester samples collected in high latitude and high altitude areas by hypothermic oriented selection. At 13℃ and 10℃, the biogas production rates of the wild low-temperature microflora bred from anaerobic environmental samples were 0.14 iwVd-m3 and 0.10mVd-m3 respectively, with methane contents of 51% and48% correspondingly. However,at the same temperatures, the digester low-temperature microflora bred from household biogas digester samples had lower biogas production rates of 0.06 tnVd-m3 and 0.04 mVd-m3 but higher methane contents of 62.0% and 56.0%. By mixing the two microflora, biogas fermentation was significantly improved with a gas production rate up to 0.15 m3/d · m3 (13 ℃) or 0.11 m3·m3 (10℃), which was much higher than that in the fermentation by either of the microflora and three times as high as that in the fermentation driven by normal temperature microflora. Biogas produced by mixed microflora contained 57.0% (13℃) or 52.0% (10℃) methane, with methane content higher than the biogas from wild low-temperature microflora, lower than that from digester low-temperature microflora, and 19.0% (13℃) or 16.0% (10℃) higher than that generated by normal temperature microflora. The results show that the mixed low-temperature biogas functional microflora is provided with excellent hypothermic biogas-producing properties and a good application prospect.%通过低温定向培养,从高纬、高寒野外厌氧环境样品和冬季沼气池底泥样品中分别选育出野生低温功能菌群和沼气池低温菌群.在13℃和10℃低温条件下,野生低温菌群产气率达到0.14 m3/d·m3和0.10 m3/d·m3,但所产沼气的甲烷含量较低,只有51.0%和48.0%.同样温度下,沼气池低温菌群的产气率只有0.06 m3/d·m3和0.04 m3/d·m3,但所产沼气的甲烷含量较高,达到62%和56%.两类菌群混合发酵可显著改善产

  4. Effects of low levels of ciprofloxacin on a chemostat model of the human colonic microflora.

    Science.gov (United States)

    Carman, R J; Woodburn, M A

    2001-06-01

    To study the utility of an in vitro model system for assessing the effect of low concentrations of a fluoroquinolone (FQ) drug on the ecology of the human intestinal microflora, chemostats containing human fecal flora were exposed to 0.43, 4.3, and 43microg of ciprofloxacin (CI) per milliliter. Prior to and during drug exposure, we assayed short-chain fatty acids (SCFA), bacterial populations, and the relative levels of susceptibility of these populations to CI and trovafloxacin (TV), a newer related FQ with increased activity against anaerobes. The degree to which CI affected the chemostat ecology was measured statistically by comparing observed data with the corresponding predicted "no effect" level. No changes in total SCFA were observed; only butyrate was significantly higher at the intermediate and high-dose levels. Enterococci counts and the levels of susceptibility to CI among enterococci were also unaffected. Escherichia coli counts decreased in a dose-dependent manner. Susceptibility levels in E. coli followed no interpretable pattern. Bacteroides fragilis group (BfG) counts decreased significantly following exposure to 43 and 4.3microg/mL CI. Ciprofloxacin susceptibility among the BfG in these chemostats was not determined because the BfG counts were too low (less than 30 colonies per plate) when undiluted chemostat samples were plated. However, within 2 days of exposure to 0.43microg/mL CI, the percentage of BfG resistant to 4microg/mL CI increased to over 95%. Before exposure, all BfG were susceptible to both CI (2microg/mL) and TV (0.25microg/mL). All BfG isolated during exposure were resistant to both CI (4microg/mL) and TV (2microg/mL). Resistance selection in the BfG was unexpected as the MIC(90) of CI for B. fragilis is 8microg/mL. Since the average colon flora is about 20% B. fragilis and other bacteroides, CI may impact the human gut flora even at subtherapeutic levels.

  5. Rhizosphere: a leverage for tolerance to water deficits of soil microflora ?

    Science.gov (United States)

    Bérard, Annette; Ruy, Stéphane; Coronel, Anaïs; Toussaint, Bruce; Czarnes, Sonia; Legendre, Laurent; Doussan, Claude

    2015-04-01

    Microbial soil communities play a fundamental role in soil organic matter mineralization, which is a key process for plant nutrition, growth and production in agro-ecosystems. A number of these microbial processes take place in the rhizosphere: the soil zone influenced by plant roots activity, which is a "hotspot " of biological and physico-chemical activity, transfers and biomass production. The knowledge of rhizosphere processes is however still scanty, especially regarding the interactions between physico-chemical processes occurring there and soil microorganisms. The rhizosphere is a place where soil aggregates are more stable, and where bulk density, porosity, water and nutrients transfer are modified with respect to the bulk soil (e.g. because of production of mucilage, of which exo-polysaccharides (EPS) produced by roots and microorganisms. During a maize field experiment, rhizospheric soil (i.e. soil strongly adhering to maize roots) and bulk soil were sampled twice in spring and summer. These soil samples were characterized for physicochemical parameters (water retention curves and analysis of exopolysaccarides) and microflora (microbial biomass, catabolic capacities of the microbial communities assessed with the MicroRespTM technique, stability of soil microbial respiration faced to a heat-drought disturbance). We observed differences between rhizospheric and bulk soils for all parameters studied: Rhizospheric soils showed higher microbial biomasses, higher quantities of exopolysaccarides and a higher water retention capacity compared to bulk soil measurements. Moreover, microbial soil respiration showed a higher stability confronted to heat-drought stress in the rhizospheric soils compared to bulk soils. Results were more pronounced during summer compared to spring. Globally these data obtained from field suggest that in a changing climate conditions, the specific physico-biological conditions in the rhizosphere partially linked to exopolysaccarides

  6. Dissemination and transmission of the E1-226V variant of chikungunya virus in Aedes albopictus are controlled at the midgut barrier level.

    Directory of Open Access Journals (Sweden)

    Camilo Arias-Goeta

    Full Text Available Emergence of arboviruses could result from their ability to exploit new environments, for example a new host. This ability is facilitated by the high mutation rate occurring during viral genome replication. The last emergence of chikungunya in the Indian Ocean region corroborates this statement since a single viral mutation at the position 226 on the E1 glycoprotein (E1-A226V was associated with enhanced transmission by the mosquito Aedes albopictus in regions where the major mosquito vector, Aedes aegypti, is absent.We used direct competition assays in vivo to dissect out the mechanisms underlying the selection of E1-226V by Ae. albopictus. When the original variant E1-226A and the newly emerged E1-226V were provided in the same blood-meal at equal titers to both species of mosquitoes, we found that the proportion of both variants was drastically different in the two mosquito species. Following ingestion of the infectious blood-meal, the E1-226V variant was preferentially selected in Ae. albopictus, whereas the E1-226A variant was sometimes favored in Ae. aegypti. Interestingly, when the two variants were introduced into the mosquitoes by intrathoracic inoculations, E1-226V was no longer favored for dissemination and transmission in Ae. albopictus, showing that the midgut barrier plays a key role in E1-226V selection.This study sheds light on the role of the midgut barrier in the selection of novel arbovirus emerging variants. We also bring new insight into how the pre-existing variant E1-226V was selected among other viral variants including E1-226A. Indeed the E1-226V variant present at low levels in natural viral populations could rapidly emerge after being selected in Ae. albopictus at the midgut barrier level.

  7. Tissue localization of the endosymbiotic bacterium "Candidatus Blochmannia floridanus" in adults and larvae of the carpenter ant Camponotus floridanus.

    Science.gov (United States)

    Sauer, Christina; Dudaczek, Dieter; Hölldobler, Bert; Gross, Roy

    2002-09-01

    The distribution of endosymbiotic bacteria in different tissues of queens, males, and workers of the carpenter ant Camponotus floridanus was investigated by light and electron microscopy and by in situ hybridization. A large number of bacteria could be detected in bacteriocytes within the midguts of workers, young virgin queens, and males. Large amounts of bacteria were also found in the oocytes of workers and queens. In contrast, bacteria were not present in oocyte-associated cells or in the spermathecae of mature queens, although occasionally a small number of bacteria could be detected in the testis follicles of males. Interestingly, the number of bacteriocytes in mature queens was strongly reduced and the bacteriocytes contained only very few or no bacteria at all, although the endosymbionts were present in huge amounts in the ovaries of the same animals. During embryogenesis of the deposited egg, the bacteria were concentrated in a ring of endodermal tissue destined to become the midgut in later developmental stages. However, during larval development, bacteria could also be detected in other tissues although to a lesser extent. Only in the last-instar larvae were bacteria found exclusively in the midgut tissue within typical bacteriocytes. Tetracycline and rifampin efficiently cleansed C. floridanus workers of their symbionts and the bacteriocytes of these animals still remained empty several months after treatment had ceased. Despite the lack of their endosymbionts, these adult animals were able to survive without any obvious negative effect under normal cultivation conditions.

  8. Effects of diet acidification and xylanase supplementation on performance, nutrient digestibility, duodenal histology and gut microflora of broilers fed wheat based diet

    NARCIS (Netherlands)

    Esmaeilipour, O.; Moravej, H.; Shivazad, M.; Rezaian, M.; Aminzadeh, S.; Krimpen, van M.M.

    2012-01-01

    1. The objective of this experiment was to study the influences of xylanase and citric acid on the performance, nutrient digestibility, digesta viscosity, duodenal histology, and gut microflora of broilers fed on a wheat based diet. 2. The experiment was carried out as a 2 x 3 factorial arrangement

  9. A comparative study on the effect of subtherapeutic tylosin administration on select feral or domestic porcine gut microflora grown in continuous-flow culture

    Science.gov (United States)

    Continuous flow cultures of feral (culture FC) and domesticated (culture RPCF) pig gut microflora were established in steady state. Cultures, in duplicate, were continuously infused subtherapeutic (25 ug/ml) levels of tylosin and sampled at intervals to assess effects on total culturable anaerobes,...

  10. The detection of subgingival microflora contents of type 2 diabetes patients with periodontitis and its correlation with gingival crevicular fluid cytokine levels

    Institute of Scientific and Technical Information of China (English)

    Ai-Hua Luo; Qian Chen; Huan Zhao; Ping Xu; Yong-Xiang Fan; Ji Shi

    2015-01-01

    Objective: To study subgingival microflora contents of type 2 diabetes patients with periodontitis and its correlation with gingival crevicular fluid cytokine levels. Methods: Type 2 diabetes patients with periodontitis and simple periodontitis patients treated in our hospital from March 2012 to August 2014 were selected for study and enrolled in diabetes with periodontitis group and simple periodontitis group respectively. Then subgingival microflora contents and levels of cytokines in gingival crevicular fluid were detected. Results: (1) Subgingival microflora: compared with simple periodontitis group, contents of porphyromonas gingivalis aeromonas, actinomycetes with haemophilus, Forsyth Tanner bacteria, middle Prairie Waugh bacteria and Treponema denticola of diabetes with periodontitis group were higher;(2) Antioxidants and oxidation products: compared with simple periodontitis group, MPO, CAT, SOD, GSH and VitC contents of diabetic with periodontitis group were lower; MDA and 8-OhdG contents were higher; (3) Signaling molecules: compared with simple periodontitis group, SFRP1, Fas, FasL, Wnt5a, NF-kb and p38 contents of diabetic with periodontitis group were higher. Conclusion: Contents of pathogens in subgingival microflora of type 2 diabetes patients with periodontitis significantly increase; antioxidants are extensively consumed and oxidation products are largely generated; there is dysfunction of multiple signaling pathways.

  11. MODULATION OF THE INTESTINAL MICROFLORA BY THE NONABSORBABLE ANTIBIOTIC VANCOMYCIN LEADS TO A REDUCTION OF THE TUMOR LOAD IN LIVER AND SPLEEN IN A LEUKEMIC RAT MODEL

    NARCIS (Netherlands)

    DAENEN, S; HUIGES, W; VEENEMA, B; VANDERWAAIJ, D; HALIE, RM

    1993-01-01

    Based on previous studies where it was shown that non-absorbable antibiotics can influence the normal hematopoiesis via changes in factors related to the intestinal microflora, the influence of vancomycin on the progression of acute myeloid leukemia was investigated in the BNML rat model. Oral vanco

  12. The effect of consumption of milk fermented by Lactobacillus casei strain Shirota on the intestinal microflora and immune parameters in humans

    NARCIS (Netherlands)

    Spanhaak, S.; Havenaar, R.; Schaafsma, G.

    1998-01-01

    Objective: To determine the effect of consumption of milk fermented by Lactobacillus casei strain Shirota (L. casei Shirota) on the composition and metabolic activities of the intestinal microflora, and immune parameters in humans. Subjects: Twenty healthy male subjects aged 40-65 years were selecte

  13. FULL-THICKNESS SMALL INTESTINE NECROSIS WITH MIDGUT VOLVULUS, DISTRIBUTED IN A PATCHY FASHION, IS REVERSIBLE WITH MODERATE BLOOD FLOW : RESUMPTION OF NORMAL FUNCTION TO NON-VIABLE INTESTINE

    OpenAIRE

    2014-01-01

    ABSTRACT Midgut volvulus is a highly life-threatening condition that carries a high risk of short gut syndrome. We report a case of catastrophic neonatal midgut volvulus in which second-look laparotomy revealed apparently non-viable remnant small intestine but with a moderate blood supply. Full-thickness small intestine necrosis was distributed in a patchy fashion, with non-viable and necrotic areas distributed so widely that no portion of the intestine could be resected. A section of full-th...

  14. Adult Strabismus

    Science.gov (United States)

    ... cause. Is eye straightening as an adult strictly cosmetic? No. Eye alignment surgery is performed in adults for several reasons. Adults ... this surgery? Eye muscle surgery is reconstructive (not cosmetic). In ... will cover strabismus surgery in adults, however, one should check with their ...

  15. ANTAGONISM OF A. VIRIDANS TO CONDITIONALLY - PATHOGENIC MICROFLORA OF THE NOSE AND OROPHARYNX OF CHILDREN WITH CARDIAC PATOLOGY

    Directory of Open Access Journals (Sweden)

    Stepansky D.O.

    2015-12-01

    Full Text Available Introduction. Search for harmless and simultaneously effective probiotics, which could be successfully used for treatment and prevention of infectious deseases, is currently important. A. viridans is of particular interest, as it is representative of the normal microflora of human with broad spectrum of antibacterial action. The use of this microorganism has a number of advantages: the absence of side effects on the body; high adhesive abilities; resistance to lysozyme in saliva; the ability of use in patients, sensitized to antibiotics and chemotherapeutic drugs; stimulation effects on the human immune system. Material and methods. The purpose of the study was to investigate the antagonism of A. viridans № 167 and autostrains of aerococcuses, isolated at patients, to conditionally - pathogenic microflora of the nose and oropharynx of children with cardiac patology. At the first stage of the study the microflora of the of the nose and oropharynx of 2 investigated categories was examined – 40 children 4-14 years with cardiac patology and 40 healthy children 4-5 years old. The second stage of work was to study the effect of A. viridans on the explored strains. Results and discussion. A. viridans manifests the antagonism to all studied strains of gram-positive and gram-negative microorganisms, except C. albisans. A. viridans antagonistic activity to staphylococci (10 + 3 mm and streptococci (10 + 2mm is at the approximately same level. It is interesting to compare the antagonism of aerococcuses to clinical isolates of S. pyogenes and similar strains from carriers (healthy children category. Impact of aerococcuses on P. mirabilis strain appeared at the highest level. Autosimbionts of A. viridans, isolated from healthy children, are more antagonistic to CPM strains, isolated from these children, than autostrains of A. viridans, isolated from children with with cardiac patology, and higher than the museum strain of A. viridans № 167 antagonism

  16. Hydrolysis of acyl-homogeneous and fish oil triacylglycerols using desalted midgut extract from atlantic salmon, Salmo salar.

    Science.gov (United States)

    Bogevik, A S; Oxley, A; Olsen, R E

    2008-07-01

    Despite several studies aimed at evaluating the positional and fatty acid specificity of fish triacylglycerol (TAG) digestive lipases, there is still much uncertainty regarding these issues. The aim of the present study was therefore to address these questions in Atlantic salmon (Salmo salar L.). Crude luminal midgut extracts were collected from fed salmon and the hydrolysis studied for various substrates including triolein (Tri-18:1), trilinolein (Tri-18:2), trilinolenin (Tri-18:3), trieicosapentaenoin (Tri-20:5), tridocosahexaenoin (Tri-22:6) and natural fish oil TAG. Using Tri-18:1, in a time-curve model showed an initial high degree of sn-1 or sn-3 specificity where sn-1,2(2,3)-diacylglycerol (1,2(2,3)-DAG) and free fatty acid (FFA) were the main hydrolytic products up to 15 min. Lack of initial sn-2 specificity was confirmed by negligible sn-1,3-diacylglycerol (1,3-DAG) being produced. During the further hydrolysis of DAG, all positions appeared susceptible to attack causing a concomitantly small increase in sn-1(3)-monoacylglycerol (1(3)-MAG) and 2-MAG, but not at the level expected for an exclusively sn-1,3-specific lipase. Oleic acid (18:1n-9) and eicosapentaenoic acid (20:5n-3) were preferred substrates for hydrolysis using both fish oil and acyl-homogeneous TAGs with FFA as the main product of lipolysis. Hydrolysis of the natural fish oil TAG appeared slower yet produced proportionally more MAG and DAG after 5 min, and similar specificities, as for synthetic TAG substrates, were exhibited with 18:1n-9 and 20:5n-3 accumulating in the FFA fraction after 30 min. Notably, 16:0 was particularly conserved in MAG. As TAG resynthesis of absorbed lipid in salmon enterocytes proceeds preferably with 2-MAG as templates, the absorption of 2-MAG, produced during initial stages of TAG hydrolysis, would need to occur rapidly to be effectively utilised via the MAG pathway.

  17. Controle químico da microflora oral em pacientes fissurados labiopalatais durante o tratamento ortodôntico-cirúrgico: estudo piloto Oral microflora chemical control in cleft lip and palate patients during orthodontic-surgical treatment pilot study

    OpenAIRE

    Alcion Alves Silva; João Luiz Carlini; Solena Ziemer Kusma

    2004-01-01

    O risco de infecção é uma preocupação durante o tratamento ortodôntico-cirúrgico, principalmente em pacientes como os fissurados lábio palatais, pois as condições anatômicas e oclusais contribuem para a proliferação microbiana. Métodos manuais para o controle da microflora oral são muitas vezes insuficientes para a obtenção de uma higiene satisfatória. Técnicas alternativas, como agentes químicos, têm sido propostas como procedimentos auxiliares. O objetivo deste estudo foi comparar a eficiên...

  18. Effects of neem oil (Azadirachta indica A. Juss) on midgut cells of predatory larvae Ceraeochrysa claveri (Navás, 1911) (Neuroptera: Chrysopidae).

    Science.gov (United States)

    Scudeler, Elton Luiz; dos Santos, Daniela Carvalho

    2013-01-01

    The effects of ingested neem oil, a botanical insecticide obtained from the seeds of the neem tree, Azadirachta indica, on the midgut cells of predatory larvae Ceraeochrysa claveri were analyzed. C. claveri were fed on eggs of Diatraea saccharalis treated with neem oil at a concentration of 0.5%, 1% and 2% during throughout the larval period. Light and electron microscopy showed severe damages in columnar cells, which had many cytoplasmic protrusions, clustering and ruptured of the microvilli, swollen cells, ruptured cells, dilatation and vesiculation of rough endoplasmic reticulum, development of smooth endoplasmic reticulum, enlargement of extracellular spaces of the basal labyrinth, intercellular spaces and necrosis. The indirect ingestion of neem oil with prey can result in severe alterations showing direct cytotoxic effects of neem oil on midgut cells of C. claveri larvae. Therefore, the safety of neem oil to non-target species as larvae of C. claveri was refuted, thus the notion that plants derived are safer to non-target species must be questioned in future ecotoxicological studies.

  19. Changes in growth and lipid profiles of silk gland, mid-gut biochemical composition of silkworm, Bombyx mori L. on exposure to prostaglandin F2alpha.

    Science.gov (United States)

    Miao, Yun-gen; Jiang, Li-jun

    2003-01-01

    The growth of the silkworm is influenced by the outside and inside environment. Among them, the category of various endocrine hormone of inside is the main factors that adjust the characters such as growth and propagate. In this experiment, we applied different dosage of prostaglandin to the fourth and fifth instar silkworm to observe the effects of prostaglandin F2alpha (PGF2alpha) on silk gland growth, mid-gut biochemical constituents and the lipid profiles of silkworm larva, Bombyx mori L. The weight of the posterior silk gland increased significantly (P lipid profiles except lipase activity suggests that the silk gland had more synthetic activity that might reflect in active spinning of silkworm larva. The changes of total proteins, free amino acids and alkaline phosphatase in mid-gut of control and PGF2alpha treated silkworm, B. mori L. indicate that PGF2alpha favored stimulatory effect on physiology of digestion, absorption and transportation of nutrients which might influence on the growth and development of larva.

  20. Apoptotic and necrotic changes in the midgut glands of the wolf spider Xerolycosa nemoralis (Lycosidae) in response to starvation and dimethoate exposure.

    Science.gov (United States)

    Wilczek, G; Rost-Roszkowska, M; Wilczek, P; Babczyńska, A; Szulińska, E; Sonakowska, L; Marek-Swędzioł, M

    2014-03-01

    In the present study, the intensity of degenerative changes (apoptosis, necrosis) in the cells of the midgut glands of male and female wolf spiders, Xerolycosa nemoralis (Lycosidae), exposed to natural (starvation) and anthropogenic (the organophosphorous pesticide dimethoate) stressors under laboratory conditions were compared. The spiders were collected from two differentially polluted sites, both located in southern Poland: Katowice-Welnowiec, which is heavily polluted with metals, and Pilica, the reference site. Starvation and dimethoate treatment resulted in enhancement of apoptotic and necrotic changes in the midgut glands of the spiders. The frequency of degenerative changes in starving individuals was twice as high as in the specimens intoxicated with dimethoate. The percentage of apoptotic and necrotic cells was higher in starving males than in starving females. A high intensity of necrotic changes, together with increased Cas-3 like activity and a greater percentage of cells with depolarized mitochondria, were typical of starving males from the polluted site. The cell death indices observed in females depended more strongly on the type of stressor than on previous preexposure to pollutants.

  1. Cadmium Accumulation and Pathological Alterations in the Midgut Gland of Terrestrial Snail Helix pomatia L. from a Zinc Smelter Area: Role of Soil pH.

    Science.gov (United States)

    Włostowski, Tadeusz; Kozłowski, Paweł; Łaszkiewicz-Tiszczenko, Barbara; Oleńska, Ewa

    2016-04-01

    The purpose of this study was to determine whether cadmium (Cd) accumulation and toxicity in the midgut gland of Helix pomatia snails living in a Cd-contaminated area were related to soil pH. Toxic responses in the midgut gland (i.e., increased vacuolization and lipid peroxidation) occurred in H. pomatia snails exhibiting the highest Cd levels in the gland (265-274 µg/g dry wt) and living on acidic soil (pH 5.3-5.5), while no toxicity was observed in snails accumulating less Cd (90 µg/g) and ranging on neutral soil (pH 7.0), despite the fact that total soil Cd was similar in the two cases. The accumulation of Cd in the gland was directly related to the water extractable Cd in soil, which in turn correlated inversely with soil pH, indicating that this factor had a significant effect on tissue Cd. It appeared further that the occurrence of Cd toxicity was associated with low levels of metallothionein in the gland of snails ranging on acidic soil.

  2. Specificity of Bacillus thuringiensis endotoxins is correlated with the presence of high-affinity binding sites in the brush border membrane of target insect midguts

    Energy Technology Data Exchange (ETDEWEB)

    Hofmann, C.; Vanderbruggen, H.; Hoefte, H.; Van Rie, J.; Jansens, S.; Van Mellaert, H. (J. Plateaustraat, Gent (Belgium))

    1988-11-01

    Binding studies were performed with two {sup 125}I-labeled Bacillus thuringiensis {delta}-endotoxins on brush border membrane vesicles prepared from the larval midgut of the tobacco hornworm Manduca sexta or the cabbage butterfly Pieris brassicae. One {delta}-endotoxin, Bt2-protoxin, is a 130-kDa recombinant crystalline protein from B. thuringiensis subsp. berliner. It kills larvae of both insect species. The active Bt2-toxin is a 60-kDa proteolytic fragment of the Bt2-protoxin. It binds saturably and with high affinity to brush border membrane vesicles from the midgut of both species. The other {delta}-endotoxin, Bt4412-protoxin, is a 136-kDa crystalline protein from B. thuringiensis subsp. thuringiensis, which is highly toxic for P. brassicae, but not for M. sexta larvae. Bt4412-toxin, obtained after proteolytic activation of Bt4412-protoxin, shows high-affinity saturable binding to P. brassicae vesicles but not to M. sexta vesicles. The correlation between toxicity and specific binding is further strengthened by competition studies. Other B. thuringiensis {delta}-endotoxins active against M. sexta compete for binding of {sup 125}I-labeled Bt2-toxin to M. sexta vesicles, whereas toxins active against dipteran or coleopteran larvae do not compete. Bt2-toxin and Bt4412-toxin bind to different sites on P. brassicae vesicles.

  3. No effect of Bt Cry1Ie toxin on bacterial diversity in the midgut of the Chinese honey bees, Apis cerana cerana (Hymenoptera, Apidae)

    Science.gov (United States)

    Jia, Hui-Ru; Dai, Ping-Li; Geng, Li-Li; Jack, Cameron J.; Li, Yun-He; Wu, Yan-Yan; Diao, Qing-Yun; Ellis, James D.

    2017-01-01

    Cry1Ie protein derived from Bacillus thuringiensis (Bt) has been proposed as a promising candidate for the development of a new Bt-maize variety to control maize pests in China. We studied the response of the midgut bacterial community of Apis cerana cerana to Cry1Ie toxin under laboratory conditions. Newly emerged bees were fed one of the following treatments for 15 and 30 days: three concentrations of Cry1Ie toxin (20 ng/mL, 200 ng/mL, and 20 μg/mL) in sugar syrup, pure sugar syrup as a negative control and 48 ng/mL imidacloprid as a positive control. The relative abundance of 16S rRNA genes was measured by Quantitative Polymerase Chain Reaction and no apparent differences were found among treatments for any of these counts at any time point. Furthermore, the midgut bacterial structure and compositions were determined using high-throughput sequencing targeting the V3-V4 regions of the 16S rDNA. All core honey bee intestinal bacterial genera such as Lactobacillus, Bifidobacterium, Snodgrassella, and Gilliamella were detected, and no significant changes were found in the species diversity and richness for any bacterial taxa among treatments at different time points. These results suggest that Cry1Ie toxin may not affect gut bacterial communities of Chinese honey bees. PMID:28139751

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

    Science.gov (United States)

    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

  5. In vivo and in vitro binding of Vip3Aa to Spodoptera frugiperda midgut and characterization of binding sites by (125)I radiolabeling.

    Science.gov (United States)

    Chakroun, Maissa; Ferré, Juan

    2014-10-01

    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 (125)I-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.

  6. Determination of the proteins encoded by BmBDV VD1-ORF4 and their interacting proteins in BmBDV-infected midguts.

    Science.gov (United States)

    Li, Guohui; Zhou, Qian; Hu, Zhaoyang; Wang, Peng; Tang, Qi; Chen, Keping; Yao, Qin

    2015-04-01

    Bombyx mori bidensovirus (BmBDV) VD1-ORF4 consists of 3,318 nucleotides, which codes for a predicted protein with molecular weight of about 127 kDa. However, the authentic proteins encoded by VD1-ORF4 in silkworm midguts infected with BmBDV and their interacting proteins are still unclear. In this study, Western blot analysis revealed that a 127-kDa protein was confirmed to be translated from the VD1-ORF4 transcript using polyclonal antibodies and monoclonal antibodies against VD1-ORF4 deduced amino acid. Moreover, four smaller proteins with molecular weight of about 70, 60, 53, and 42 kDa were also examined in the infected midguts. Transient expression assay indicated that the expression amount of VD1-ORF4 fused with egfp was at least 30-fold lower than that of egfp gene, and immunofluorescence staining result indicated that these proteins encoded by VD1-ORF4 were located in both the cytoplasm and nucleus. Co-immunoprecipitation result showed that Aminopeptidase and Heat shock protein 90 can be captured by these proteins encoded by VD1-ORF4. In conclusion, multiple proteins were produced from the transcripts of VD1-ORF4 gene by an uncertain expression strategy, which may play important roles in viral replication and assembly.

  7. Effects of different brush border membrane vesicle isolation protocols on proteomic analysis of Cry1 Ac binding proteins from the midgut of Helicoverpa armigera

    Institute of Scientific and Technical Information of China (English)

    Li-Zhen Chen; Ge-Mei Liang; Brian G.Rector; Jie Zhang; Kong-Ming Wu; Yu-Yuan Guo

    2008-01-01

    Brush border membrane vesicles(BBMV)isolated from insect midguts have been widely used to study CrylA binding proteins.Sample preparation is important in two-dimensional electrophoresis(2-DE),so to determine a suitable BBMV preparation method in Helicoverpa armigera for 2-DE,we compared three published BBMV preparation methods mostly used in sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE).All mctllods yielded similar types and numbers of binding proteins,but in different quantifies.The Abdul.Rauf and Ellar protocol was the best of the three,but had limitations.Sufficient protein qu antity iS important for research involving limited numbers of insects,such as studies of insect resistance to Bacillus thuringiensis in the field.Consequently,we integrated the three BBMV isolation methods into a single protocol that yielded high quantities of BBMV proteins from H.armigera larval midguts.which proved suitable for 2-DE analysis.

  8. Ferritin acts as a target site for the snowdrop lectin(GNA)in the midgut of the cotton leafworm Spodoptera fittoralis

    Institute of Scientific and Technical Information of China (English)

    Amin Sadeghi; Guy Smagghe; Paul Proost; EIs J.M.Van Damme

    2008-01-01

    The snowdrop lcctin GNA(Galanthus nivalis agglutininl has been shown to possess insecticidal activity to a range of economically important insect pcsts.However,the precise mechanism of insecticidal action of GNA against insects reinains unknown.In this investigation,we attempted to purify and identify receptor(s)responsible for binding of GNA in the larval midgut of a major lepidopteran pest(the cotton leafworm,Spodoptera littoralis) to better understand its mode of action.Therefore,cytoplasmic as well as membrane proteins from 800 larval midguts were chromatographed on a column with immobilized GNA.Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of the proteins eluted from the GNA column followed by sequencing of the GNA-binding proteins and BLAST analyses revealed that the N-terminal sequences of a 24 kDa polypep-tide purified from the cytoplasmic and membrane protein fraction revealed sequence similarity to sequences encoding heavy chain homologs of ferritin from Manduca sexta (76%sequence identity),Carpodes ethlius(80%sequence identity)and Bombyx mori(61%sequence identity).Furthermore,the N-terminal sequence of a 31 kDa polypeptide from the membrane protein fraction showed sequence similarity to a light chain homolog of ferritin from Manduca sexta(88%sequence identity).

  9. Effect of blood components, abdominal distension, and ecdysone therapy on the ultrastructural organization of posterior midgut epithelial cells and perimicrovillar membranes in Rhodnius prolixus

    Directory of Open Access Journals (Sweden)

    JM Albuquerque-Cunha

    2004-12-01

    Full Text Available The effects of blood components, nerve-cord severance, and ecdysone therapy on the posterior midgut epithelial cells of 5th-instar Rhodnius prolixus nymphs 10 days after feeding were analyzed by transmission electron microscopy. Cutting the nerve-cord of the blood-fed insects partially reduced the development of microvilli and perimicrovillar membranes (PMM, and produced large vacuoles and small electrondense granules; insects fed on Ringer's saline diet exhibited well developed microvilli and low PMM production; swolled rough endoplasmatic reticulum and electrondense granules; Ringer's saline meal with ecdysone led to PMM development, glycogen particles, and several mitochondria in the cytoplasm; epithelial cells of the insects fed on Ringer's saline meal whose nerve-cord was severed showed heterogeneously distributed microvilli with reduced PMM production and a great quantity of mitochondria and glycogen in the cytoplasm; well developed microvilli and PMM were observed in nerve-cord severed insects fed on Ringer's saline meal with ecdysone; Ringer's saline diet containing hemoglobin recovered the release of PMM; and insects fed on human plasma showed slightly reduced PMM production, although the addition of ecdysone in the plasma led to a normal midgut ultrastructural organization. We suggest that the full development of microvilli and PMM in the epithelial cells depends on the abdominal distension in addition to ingestion of hemoglobin, and the release of ecdysone.

  10. Immunology and probiotic impact of the newborn and young children intestinal microflora.

    Science.gov (United States)

    Bezirtzoglou, Eugenia; Stavropoulou, Elisabeth

    2011-12-01

    Human body has developed a holistic defence system, which mission is either to recognize and destroy the aggressive invaders or to evolve mechanisms permitting to minimize or restore the consequences of harmful actions. The host immune system keeps the capital role to preserve the microbial intestinal balance via the barrier effect. Specifically, pathogenic invaders such as, bacteria, parasites, viruses and other xenobiotic invaders are rejected out of the body via barriers formed by the skin, mucosa and intestinal flora. In case physical barriers are breached, the immune system with its many components comes into action in order to fence infection. The intestine itself is considered as an "active organ" due to its abundant bacterial flora and to its large metabolic activity. The variation among different species or even among different strains within a species reflects the complexity of the genetic polymorphism which regulates the immune system functions. Additionally factors such as, gender, particular habits, smoking, alcohol consumption, diet, religion, age, gender, precedent infections and vaccinations must be involved. Hormonal profile and stress seems to be associated to the integrity microbiota and inducing immune system alterations. Which bacterial species are needed for inducing a proper barrier effect is not known, but it is generally accepted that this barrier function can be strongly supported by providing benefic alimentary supplements called functional foods. In this vein it is stressed the fact that early intestinal colonization with organisms such as Lactobacilli and Bifidobacteria and possibly subsequent protection from many different types of diseases. Moreover, this benefic microflora dominated but Bifidobacteria and Lactobacilli support the concept of their ability to modify the gut microbiota by reducing the risk of cancer following their capacity to decrease β-glucoronidase and carcinogen levels. Because of their beneficial roles in the

  11. Adult Education and Adult Learning

    DEFF Research Database (Denmark)

    Illeris, Knud

    Kort beskrivelse Bogen, 'Adult Education og Adult Learning', giver et fyldestgørende overblik over forståelsen af voksenuddannelse og læring. Abstract I "Adult Education and Adult Learning' ser Knud Illeris på voksenuddannelse fra to perspektiver. På den ene side beskrives de aktuelle udfordringer...

  12. Impact of probiotics on colonic microflora in patients with colitis: A prospective double blind randomised crossover study

    DEFF Research Database (Denmark)

    Ahmed, Jamil; Reddy, Bala S.; Mølbak, Lars;

    2013-01-01

    for a month and then “crossed over” to receive a placebo, or alternatively to receive the placebo first followed in the second month by synbiotic. Stool samples were collected on entry into the study and then at the end of first and second months respectively. Colonic microflora was measured by terminal...... restriction fragment length polymorphism technique. Quantitative PCR was used to determine the concentration of individual species. ResultsSixteen patients completed the study of whom 8 had Crohn's colitis and 8 had ulcerative colitis. Their median age was 62 (IQR 50–65) years. An average of 22 terminal...... restriction fragments (T-RF's) was identified in each patient. Dice cluster analysis showed that each patient had a unique microbial composition which did not change significantly at different time points in the study, irrespective of whether they had probiotics or the placebo. Probiotic organisms were...

  13. The role of conditionally pathogenic microflora in the immune pathologic reactions development at Epstein-Barr virus infection

    Directory of Open Access Journals (Sweden)

    Marina Kim

    2014-04-01

    Full Text Available The present article is concerned with studying of appoptogenic activity of microbes associated with Epstein-Barr virus infection on model of peritoneal mice macrophages in vitro. The estimation of apoptosis induced by bacterias extracted from patients with Epstein-Barr virus infection was carried out oncharacteristic morphologic changes of macrophages. It was established that apoptogenic activity had all microbes associated with Epstein-Barr virus infection, but the highiest pathogenic potential Streptococcus pyogenes. Presence of apoptogenic activity of bacterial microflora associated with Epstein-Barr virus infection towards immune system cells can serve as pathway of their survival and can be the pathogenic base of their long persistence in the body. The expressed ability of Streptococcus pyogenes to induce the apoptosis can give us the possibility to consider such microbe as the most dangerous microbe associated with Epstein-Barr virus infection.

  14. Fermentation and microflora of plaa-som, a Thai fermented fish product prepared with different salt concentrations

    DEFF Research Database (Denmark)

    Paludan-Müller, Christine; Madsen, M.; Sophanodora, P.

    2002-01-01

    Plaa-som is a Thai fermented fish product prepared from snakehead fish, salt, palm syrup and sometimes roasted rice. We studied the effects of different salt concentrations on decrease in pH and on microflora composition during fermentation. Two low-salt batches were prepared, containing 6% and 7......% salt (w/w) as well as two high-salt batches, containing 9% and 11% salt. pH decreased rapidly from 6 to 4.5 in low-salt batches, whereas in high-salt batches, a slow or no decrease in pH was found. Lactic acid bacteria (LAB) and yeasts were isolated as the dominant microorganisms during fermentation...

  15. The effect of Lacto- and Bifidobacteria in monoculture on the vaginal microflora in norm and in cases of intravaginal staphylococcosis.

    Science.gov (United States)

    Babenko, L R; Lazarenko, L M; Shynkarenko, L M; Mokrozub, V V; Pidgorskyi, V S; Spivak, M Ja

    2013-01-01

    The influence of probiotic strains of Lactobacillus casei IMV B-7280, Lactobacillus acidophilus IMV B-7279, Bifidobacterium animalis VKL, Lactobacillus delbrueckii subsp. bulgaricus IMV B-7281 and Bifidobacterium animalis VKB, each strain separately, to the range of the urogenital tract microflora in physiological norm and in cases of experimental intravaginal staphylococcosis of mice induced by Staphylococcus aureus 8325-4. It was found that all these strains had different efficiency in Staphylococcus aureus 8325-4 growth suppression in the vagina of infected mice and affect the spectrum of microorganisms. Lactobacillus casei IMV B-7280 strain had effective antistaphylococcal effect and reduced the number of coliform bacteria and fungi in the vagina in cases of intravaginal staphylococcosis of mice. Lactobacillus casei IMV B-7280 strain is promising to create probiotic drugs effective in treating intravaginal staphylococcosis.

  16. Single versus repeated applications of CuO and Ag nanomaterials and their effect on soil microflora.

    Science.gov (United States)

    Schlich, Karsten; Beule, Lukas; Hund-Rinke, Kerstin

    2016-08-01

    Nanomaterials enter the terrestrial environment via the repeated application of sludge to soils over many years. The goal of this investigation was to compare the effects of CuO and Ag nanomaterials on soil microorganisms after a single application and after repeated applications ultimately resulting in the same test concentrations. The effect on soil microorganisms was determined using the ammonium oxidation (ISO 15685), enzymatic activity patterns (ISO 22939) and MicroResp™ tests on days 28, 56 and 84. The comparability of single and repeated applications of ion-releasing nanomaterials depended on the test endpoint and duration. No significant differences between single and repeated applications were observed when testing nitrifying microorganisms and exoenzymes, but differences were observed in the substrate-induced respiration test. The three test systems used together provide more comprehensive information about the impact of different nanomaterials on the soil microflora and its diversity.

  17. Identification of urolithin a as a metabolite produced by human colon microflora from ellagic acid and related compounds.

    Science.gov (United States)

    Cerdá, Begoña; Periago, Paula; Espín, Juan Carlos; Tomás-Barberán, Francisco A

    2005-07-13

    Dietary ellagic acid and related polyphenols are metabolized in humans to dibenzopyran-6-one derivatives, and the microbial origin of these metabolites has been suggested. However, this has not been demonstrated so far. Fecal samples donated by six volunteers were incubated under anaerobic conditions, and aliquots were used to evaluate the fecal metabolism of ellagic acid, the ellagitannin punicalagin, and an ellagitannin rich extract from walnuts. The isoflavone daidzein was also incubated with the same fecal samples to follow the production of the microbial metabolites previously reported (dihydrogenistein, O-demethylangolensin, and equol) as a positive control of the system and to evaluate similarities between isoflavone and ellagic acid fecal flora metabolism. After fermentation the metabolite "urolithin A" (3,8-dihydroxy-6H-dibenzo[b,d]pyran-6-one) was produced from ellagic acid, punicalagin, and the ellagitannin extract in all the fecal cultures from different volunteers, but with very different production rates and concentrations. This large variability in the concentration of metabolite and kinetics of metabolite production is consistent with the large variability found in the excretion of these metabolites in urine in vivo after human consumption of ellagitannins, and with differences in the composition of the fecal microflora. No correlation between isoflavone and ellagic acid metabolism by fecal microflora was observed. The present study confirms the microbial origin of the recently reported in vivo generated hydroxy-6H-dibenzo[b,d]pyran-6-one derivatives in humans and is a further step in the study of the bioavailability and metabolism of ellagic acid and ellagitannins.

  18. Supplementation of milk formula with galacto-oligosaccharides improves intestinal micro-flora and fermentation in term infants

    Institute of Scientific and Technical Information of China (English)

    BEN Xiao-ming 贲晓明; ZHOU Xiao-yu 周晓玉; ZHAO Wei-hua 赵卫华; YU Wen-liang 喻文亮; PAN Wei 潘伟; ZHANG Wei-li 张伟利; WU Sheng-mei 吴圣楣; Christien M.Van Beusekom; Anne Schaafsma

    2004-01-01

    Background Oligosaccharides in human milk may protect infants by improving the intestinal micro-flora and fermentation. This study was to investigate effects of infant formula milk consisting of galacto-oligosaccharide (GOS) on intestinal microbial populations and the fermentation characteristics in term infants in comparison with that of human milk. Methods The test formula (Frisolac H, Friesland, Netherland) was supplemented with GOS at a concentration of 0.24 g/dl. Human milk and another formula without oligosaccharides (Frisolac H, Friesland, Netherland) were used as positive and negative control respectively. Growth, stool characteristics, and side effects of the recruited infants were recorded after 3 and 6 months' follow-up, and the fecal species were collected for the analysis of intestinal micro-flora, short chain fatty acid (SCFA) and pH.Results At the end of 3- and 6-month feeding period, intestinal Bifidobacteria and Lactobacilli were significantly increased in infants fed with GOS supplemented formula and human milk when compared with infants fed with negative control formula; however, there was no statistically significant difference between GOS supplemented formula and human milk groups. Stool characteristics were influenced by the supplement and main fecal SCFA (acetic), and stool frequency were significantly increased in infants fed with GOS supplemented formula and human milk, while the fecal pH was significantly decreased as compared with that of negative control (P<0.05). Supplementation had no influence on incidence of side effects (including crying, regurgitation and vomiting). Conclusions Supplementing infant formula with GOS at a concentration of 0.24 g/dl stimulates the growth of Bifidobacteria and Lactobacilli in the intestine and stool characteristics are similar to in term infants fed with human milk.

  19. Study on Main Microflora of Yanghe Daqu%洋河大曲主要菌系的研究

    Institute of Scientific and Technical Information of China (English)

    周新虎; 陈翔; 李燕荣; 葛向阳

    2011-01-01

    对洋河大由3种大曲浸提液主体微生物区系进行研究。结果表明,洋河大曲中的细菌种类最为丰富,共分离到15株,其中芽孢杆菌属6株;霉菌12株,其中,曲霉属6株,该种类较丰富,但其在数量上并不具优势;分离到酵母菌8株,其中,毕赤酵母属3株,其数量占酵母菌总数的80%左右;洋河大曲与传统浓香型大曲中的主体微生物菌系之间存在一定的差异性。%Three kinds of extract solution of Yanghe Daqu were selected to investigate their main microflora. The results showed that there were rich bacterial species (15 strains isolated including 6 Bacillus strains), there isolated 12 mildew strains including 6 Aspergillus strains but their quantity was not predominant, and there isolated 8 yeast strains including 3 P/ch/a strains (their quantity accounted for 80 % of total yeast quantity). As a result from the above studies, it was found that the difference of main microflora between Yanghe Daqu and traditional Luzhou-flavor Daqu-was evident.

  20. Polymyositis - adult

    Science.gov (United States)

    ... rash is a sign of a similar condition, dermatomyositis . Common symptoms include: Muscle weakness in the shoulders ... in the treatment of refractory adult and juvenile dermatomyositis and adult polymyositis: a randomized, placebo-phase trial. ...

  1. Different Effects of Bacillus thuringiensis Toxin Cry1Ab on Midgut Cell Transmembrane Potential of Mythimna separata and Agrotis ipsilon Larvae

    Science.gov (United States)

    Wang, Yingying; Hu, Zhaonong; Wu, Wenjun

    2015-01-01

    Bacillus thuringiensis (Bt) Cry toxins from the Cry1A family demonstrate significantly different toxicities against members of the family Noctuidae for unknown reasons. In this study, membrane potential was measured and analyzed in freshly isolated midgut samples from Mythimna separata and Agrotis ipsilon larvae under oral administration and in vitro incubation with Bt toxin Cry1Ab to elucidate the mechanism of action for further control of these pests. Bioassay results showed that the larvae of M. separata achieved a LD50 of 258.84 ng/larva at 24 h after ingestion; M. separata larvae were at least eightfold more sensitive than A. ipsilon larvae to Cry1Ab. Force-feeding showed that the observed midgut apical-membrane potential (Vam) of M. separata larvae was significantly depolarized from −82.9 ± 6.6 mV to −19.9 ± 7.2 mV at 8 h after ingestion of 1 μg activated Cry1Ab, whereas no obvious changes were detected in A. ipsilon larvae with dosage of 5 μg Cry1Ab. The activated Cry1Ab caused a distinct concentration-dependent depolarization of the apical membrane; Vam was reduced by 50% after 14.7 ± 0.2, 9.8 ± 0.4, and 7.6 ± 0.6 min of treatment with 1, 5, and 10 μg/mL Cry1Ab, respectively. Cry1Ab showed a minimal effect on A. ipsilon larvae even at 20 μg/mL, and Vam decreased by 26.3% ± 2.3% after 15 min. The concentrations of Cry1Ab displayed no significant effect on the basolateral side of the epithelium. The Vam of A. ipsilon (−33.19 ± 6.29 mV, n = 51) was only half that of M. separata (−80.94 ± 6.95 mV, n = 75). The different degrees of sensitivity to Cry1Ab were speculatively associated with various habits, as well as the diverse physiological or biochemical characteristics of the midgut cell membranes. PMID:26694463

  2. Different Effects of Bacillus thuringiensis Toxin Cry1Ab on Midgut Cell Transmembrane Potential of Mythimna separata and Agrotis ipsilon Larvae

    Directory of Open Access Journals (Sweden)

    Yingying Wang

    2015-12-01

    Full Text Available Bacillus thuringiensis (Bt Cry toxins from the Cry1A family demonstrate significantly different toxicities against members of the family Noctuidae for unknown reasons. In this study, membrane potential was measured and analyzed in freshly isolated midgut samples from Mythimna separata and Agrotis ipsilon larvae under oral administration and in vitro incubation with Bt toxin Cry1Ab to elucidate the mechanism of action for further control of these pests. Bioassay results showed that the larvae of M. separata achieved a LD50 of 258.84 ng/larva at 24 h after ingestion; M. separata larvae were at least eightfold more sensitive than A. ipsilon larvae to Cry1Ab. Force-feeding showed that the observed midgut apical-membrane potential (Vam of M. separata larvae was significantly depolarized from −82.9 ± 6.6 mV to −19.9 ± 7.2 mV at 8 h after ingestion of 1 μg activated Cry1Ab, whereas no obvious changes were detected in A. ipsilon larvae with dosage of 5 μg Cry1Ab. The activated Cry1Ab caused a distinct concentration-dependent depolarization of the apical membrane; Vam was reduced by 50% after 14.7 ± 0.2, 9.8 ± 0.4, and 7.6 ± 0.6 min of treatment with 1, 5, and 10 μg/mL Cry1Ab, respectively. Cry1Ab showed a minimal effect on A. ipsilon larvae even at 20 μg/mL, and Vam decreased by 26.3% ± 2.3% after 15 min. The concentrations of Cry1Ab displayed no significant effect on the basolateral side of the epithelium. The Vam of A. ipsilon (−33.19 ± 6.29 mV, n = 51 was only half that of M. separata (−80.94 ± 6.95 mV, n = 75. The different degrees of sensitivity to Cry1Ab were speculatively associated with various habits, as well as the diverse physiological or biochemical characteristics of the midgut cell membranes.

  3. Identification of different trypanosome species in the mid-guts of tsetse flies of the Malanga (Kimpese sleeping sickness focus of the Democratic Republic of Congo

    Directory of Open Access Journals (Sweden)

    Simo Gustave

    2012-09-01

    Full Text Available Abstract Background The Malanga sleeping sickness focus of the Democratic Republic of Congo has shown an epidemic evolution of disease during the last century. However, following case detection and treatment, the prevalence of the disease decreased considerably. No active survey has been undertaken in this focus for a couple of years. To understand the current epidemiological status of sleeping sickness as well as the animal African trypanosomiasis in the Malanga focus, we undertook the identification of tsetse blood meals as well as different trypanosome species in flies trapped in this focus. Methods Pyramidal traps were use to trap tsetse flies. All flies caught were identified and live flies were dissected and their mid-guts collected. Fly mid-gut was used for the molecular identification of the blood meal source, as well as for the presence of different trypanosome species. Results About 949 Glossina palpalis palpalis were trapped; 296 (31.2% of which were dissected, 60 (20.3% blood meals collected and 57 (19.3% trypanosome infections identified. The infection rates were 13.4%, 5.1%, 3.5% and 0.4% for Trypanosoma congolense savannah type, Trypanosoma brucei s.l., Trypanosoma congolense forest type and Trypanosoma vivax, respectively. Three mixed infections including Trypanosoma brucei s.l. and Trypanosoma congolense savannah type, and one mixed infection of Trypanosoma vivax and Trypanosoma congolense savannah type were identified. Eleven Trypanosoma brucei gambiense infections were identified; indicating an active circulation of this trypanosome subspecies. Of all the identified blood meals, about 58.3% were identified as being taken on pigs, while 33.3% and 8.3% were from man and other mammals, respectively. Conclusion The presence of Trypanosoma brucei in tsetse mid-guts associated with human blood meals is indicative of an active transmission of this parasite between tsetse and man. The considerable number of pig blood meals combined

  4. Study of the Bacillus thuringiensis Vip3Aa16 histopathological effects and determination of its putative binding proteins in the midgut of Spodoptera littoralis.

    Science.gov (United States)

    Abdelkefi-Mesrati, Lobna; Boukedi, Hanen; Dammak-Karray, Mariam; Sellami-Boudawara, Tahya; Jaoua, Samir; Tounsi, Slim

    2011-02-01

    The bacterium Bacillus thuringiensis produces, at the vegetative stage of its growth, Vip3A proteins with activity against a broad spectrum of lepidopteran insects. The Egyptian cotton leaf worm (Spodoptera littoralis) is an important agricultural pest that is susceptible to the Vip3Aa16 protein of Bacillus thuringiensis kurstaki strain BUPM95. The midgut histopathology of Vip3Aa fed larvae showed vacuolization of the cytoplasm, brush border membrane destruction, vesicle formation in the apical region and cellular disintegration. Biotinylated Vip3Aa toxin bound proteins of 55- and 100-kDa on blots of S. littoralis brush border membrane preparations. These binding proteins differ in molecular size from those recognized by Cry1C, one of the very few Cry proteins active against the polyphagous S. littoralis. This result supports the use of Vip3Aa16 proteins as insecticidal agent, especially in case of Cry-resistance management.

  5. The impact of the Bacillus subtilis SPB1 biosurfactant on the midgut