Sample records for jejuni boiled cell

  1. Invasion of epithelial cells by Campylobacter jejuni is independent of caveolae

    PubMed Central

    2013-01-01

    Caveolae are 25–100 nm flask-like membrane structures enriched in cholesterol and glycosphingolipids. Researchers have proposed that Campylobacter jejuni require caveolae for cell invasion based on the finding that treatment of cells with the cholesterol-depleting compounds filipin III or methyl-β-cyclodextrin (MβCD) block bacterial internalization in a dose-dependent manner. The purpose of this study was to determine the role of caveolae and caveolin-1, a principal component of caveolae, in C. jejuni internalization. Consistent with previous work, we found that the treatment of HeLa cells with MβCD inhibited C. jejuni internalization. However, we also found that the treatment of HeLa cells with caveolin-1 siRNA, which resulted in greater than a 90% knockdown in caveolin-1 protein levels, had no effect on C. jejuni internalization. Based on this observation we performed a series of experiments that demonstrate that MβCD acts broadly, disrupting host cell lipid rafts and C. jejuni-induced cell signaling. More specifically, we found that MβCD inhibits the cellular events necessary for C. jejuni internalization, including membrane ruffling and Rac1 GTPase activation. We also demonstrate that MβCD disrupted the association of the β1 integrin and EGF receptor, which are required for the maximal invasion of epithelial cells. In agreement with these findings, C. jejuni were able to invade human Caco-2 cells, which are devoid of caveolae, at a level equal to that of HeLa cells. Taken together, the results of our study demonstrate that C. jejuni internalization occurs in a caveolae-independent manner. PMID:24364863

  2. Polynucleotide phosphorylase has an impact on cell biology of Campylobacter jejuni

    PubMed Central

    Haddad, Nabila; Tresse, Odile; Rivoal, Katell; Chevret, Didier; Nonglaton, Quentin; Burns, Christopher M.; Prévost, Hervé; Cappelier, Jean M.

    2012-01-01

    Polynucleotide phosphorylase (PNPase), encoded by the pnp gene, is known to degrade mRNA, mediating post-transcriptional regulation and may affect cellular functions. The role of PNPase is pleiotropic. As orthologs of the two major ribonucleases (RNase E and RNase II) of Escherichia coli are missing in the Campylobacter jejuni genome, in the current study the focus has been on the C. jejuni ortholog of PNPase. The effect of PNPase mutation on C. jejuni phenotypes and proteome was investigated. The inactivation of the pnp gene reduced significantly the ability of C. jejuni to adhere and to invade Ht-29 cells. Moreover, the pnp mutant strain exhibited a decrease in C. jejuni swimming ability and chick colonization. To explain effects of PNPase on C. jejuni 81-176 phenotype, the proteome of the pnp mutant and parental strains were compared. Overall, little variation in protein production was observed. Despite the predicted role of PNPase in mRNA regulation, the pnp mutation did not induce profound proteomic changes suggesting that other ribonucleases in C. jejuni might ensure this biological function in the absence of PNPase. Nevertheless, synthesis of proteins which are involved in virulence (LuxS, PEB3), motility (N-acetylneuraminic acid synthetase), stress-response (KatA, DnaK, Hsp90), and translation system (EF-Tu, EF-G) were modified in the pnp mutant strain suggesting a more specific role of PNPase in C. jejuni. In conclusion, PNPase deficiency induces limited but important consequences on C. jejuni biology that could explain swimming limitation, chick colonization delay, and the decrease of cell adhesion/invasion ability. PMID:22919622

  3. Campylobacter jejuni induces transcytosis of commensal bacteria across the intestinal epithelium through M-like cells

    PubMed Central

    2010-01-01

    Background Recent epidemiological analyses have implicated acute Campylobacter enteritis as a factor that may incite or exacerbate inflammatory bowel disease (IBD) in susceptible individuals. We have demonstrated previously that C. jejuni disrupts the intestinal barrier function by rapidly inducing epithelial translocation of non-invasive commensal bacteria via a transcellular lipid raft-mediated mechanism ('transcytosis'). To further characterize this mechanism, the aim of this current study was to elucidate whether C. jejuni utilizes M cells to facilitate transcytosis of commensal intestinal bacteria. Results C. jejuni induced translocation of non-invasive E. coli across confluent Caco-2 epithelial monolayers in the absence of disrupted transepithelial electrical resistance or increased permeability to a 3 kDa dextran probe. C. jejuni-infected monolayers displayed increased numbers of cells expressing the M cell-specific marker, galectin-9, reduced numbers of enterocytes that stained with the absorptive enterocyte marker, Ulex europaeus agglutinin-1, and reduced activities of enzymes typically associated with absorptive enterocytes (namely alkaline phosphatase, lactase, and sucrase). Furthermore, in Campylobacter-infected monolayers, E. coli were observed to be internalized specifically within epithelial cells displaying M-like cell characteristics. Conclusion These data indicate that C. jejuni may utilize M cells to promote transcytosis of non-invasive bacteria across the intact intestinal epithelial barrier. This mechanism may contribute to the inflammatory immune responses against commensal intestinal bacteria commonly observed in IBD patients. PMID:21040540

  4. An adapted in vitro assay to assess Campylobacter jejuni interaction with intestinal epithelial cells: Taking into stimulation with TNFα.

    PubMed

    Rodrigues, Ramila Cristiane; Pocheron, Anne-Lise; Cappelier, Jean-Michel; Tresse, Odile; Haddad, Nabila

    2018-06-01

    Campylobacter jejuni is the most prevalent foodborne bacterial infection agent. This pathogen seems also involved in inflammatory bowel diseases in which pro-inflammatory cytokines, such as tumor necrosis factor α (TNFα), play a major role. C. jejuni pathogenicity has been extensively studied using in vitro cell culture methods, and more precisely "healthy" cells. In fact, no information is available regarding the behavior of C. jejuni in contact with TNFα-stimulated cells. Therefore, this research was designed to investigate the effect of TNFα on C. jejuni interaction with human intestinal epithelial cells (HT29 and HT29-MTX). To ensure IL-8 production induced by TNFα, human rtTNFα was added to HT29 and HT29-MTX before adhesion and invasion assays. About 10 8 CFU bacteria of C. jejuni strains cells were added to measure their adherence and invasion abilities using TNFα-stimulated cells versus non stimulated cells. Exposure to TNFα results in IL-8 overproduction by intestinal epithelial cells. In addition, the effect of TNFα pre-treatment on C. jejuni adhesion and internalization into eukaryotic cells is strain-dependent. Indeed, the adhesion/invasion process is affected in <50% of the strains tested when TNFα is added to the intestinal cells. Interestingly, TNFα affects more strains in their ability to adhere to and invade the mucus-secreting HT29-MTX cells. Among the 10 strains tested, the aero-tolerant C. jejuni Bf strain is one of the most virulent. These results suggest that the TNFα signalling pathway could participate in the internalization of C. jejuni in human intestinal cells and can help in understanding the pathogenicity of this microorganism in contact with TNFα-stimulated cells. Copyright © 2018. Published by Elsevier B.V.

  5. Campylobacter fetus subsp. jejuni in poultry reared under different management systems in Nigeria.

    PubMed

    Adekeye, J O; Abdu, P A; Bawa, E K

    1989-01-01

    Cloacal swabs from 487 live birds in 36 flocks and 70 poultry carcasses were cultured for Campylobacter fetus subsp. jejuni. It was isolated from 12.3% of the birds in 19 flocks. Chickens, turkeys, and guinea fowl differed from one another in isolation rates of the organism. Management system affected its occurrence, and only 7.1% of eviscerated carcasses yielded it. It was concluded that bird species, management system, and immersing slaughtered poultry in boiling water before dressing affect recovery of C. fetus subsp. jejuni from live birds and carcasses.

  6. In Vitro Evaluation of the Impact of the Probiotic E. coli Nissle 1917 on Campylobacter jejuni's Invasion and Intracellular Survival in Human Colonic Cells.

    PubMed

    Helmy, Yosra A; Kassem, Issmat I; Kumar, Anand; Rajashekara, Gireesh

    2017-01-01

    Campylobacter jejuni is a leading cause of bacterial food poisoning in humans. Due to the rise in antibiotic-resistant Campylobacter , there exists a need to develop antibiotic-independent interventions to control infections in humans. Here, we evaluated the impact of Escherichia coli Nissle 1917 (EcN), a probiotic strain, on C. jejuni's invasion and intracellular survival in polarized human colonic cells (HT-29). To further understand how EcN mediates its impact, the expression of 84 genes associated with tight junctions and cell adhesion was profiled in HT-29 cells after treatment with EcN and challenge with C. jejuni . The pre-treatment of polarized HT-29 cells with EcN for 4 h showed a significant effect on C. jejuni 's invasion (∼2 log reduction) of the colonic cells. Furthermore, no intracellular C. jejuni were recovered from EcN pre-treated HT-29 cells at 24 h post-infection. Other probiotic strains tested had no significant impact on C. jejuni invasion and intracellular survival. C. jejuni decreased the expression of genes associated with epithelial cells permeability and barrier function in untreated HT-29 cells. However, EcN positively affected the expression of genes that are involved in enhanced intestinal barrier function, decreased cell permeability, and increased tight junction integrity. The results suggest that EcN impedes C. jejuni invasion and subsequent intracellular survival by affecting HT-29 cells barrier function and tight junction integrity. We conclude that EcN might be a viable alternative for controlling C. jejuni infections.

  7. Detection of Small Numbers of Campylobacter jejuni and Campylobacter coli Cells in Environmental Water, Sewage, and Food Samples by a Seminested PCR Assay

    PubMed Central

    Waage, Astrid S.; Vardund, Traute; Lund, Vidar; Kapperud, Georg

    1999-01-01

    A rapid and sensitive assay was developed for detection of small numbers of Campylobacter jejuni and Campylobacter coli cells in environmental water, sewage, and food samples. Water and sewage samples were filtered, and the filters were enriched overnight in a nonselective medium. The enrichment cultures were prepared for PCR by a rapid and simple procedure consisting of centrifugation, proteinase K treatment, and boiling. A seminested PCR based on specific amplification of the intergenic sequence between the two Campylobacter flagellin genes, flaA and flaB, was performed, and the PCR products were visualized by agarose gel electrophoresis. The assay allowed us to detect 3 to 15 CFU of C. jejuni per 100 ml in water samples containing a background flora consisting of up to 8,700 heterotrophic organisms per ml and 10,000 CFU of coliform bacteria per 100 ml. Dilution of the enriched cultures 1:10 with sterile broth prior to the PCR was sometimes necessary to obtain positive results. The assay was also conducted with food samples analyzed with or without overnight enrichment. As few as ≤3 CFU per g of food could be detected with samples subjected to overnight enrichment, while variable results were obtained for samples analyzed without prior enrichment. This rapid and sensitive nested PCR assay provides a useful tool for specific detection of C. jejuni or C. coli in drinking water, as well as environmental water, sewage, and food samples containing high levels of background organisms. PMID:10103261

  8. Heat Shock-Enhanced Conjugation Efficiency in Standard Campylobacter jejuni Strains

    PubMed Central

    Zeng, Ximin; Ardeshna, Devarshi

    2015-01-01

    Campylobacter jejuni, the leading bacterial cause of human gastroenteritis in the United States, displays significant strain diversity due to horizontal gene transfer. Conjugation is an important horizontal gene transfer mechanism contributing to the evolution of bacterial pathogenesis and antimicrobial resistance. It has been observed that heat shock could increase transformation efficiency in some bacteria. In this study, the effect of heat shock on C. jejuni conjugation efficiency and the underlying mechanisms were examined. With a modified Escherichia coli donor strain, different C. jejuni recipient strains displayed significant variation in conjugation efficiency ranging from 6.2 × 10−8 to 6.0 × 10−3 CFU per recipient cell. Despite reduced viability, heat shock of standard C. jejuni NCTC 11168 and 81-176 strains (e.g., 48 to 54°C for 30 to 60 min) could dramatically enhance C. jejuni conjugation efficiency up to 1,000-fold. The phenotype of the heat shock-enhanced conjugation in C. jejuni recipient cells could be sustained for at least 9 h. Filtered supernatant from the heat shock-treated C. jejuni cells could not enhance conjugation efficiency, which suggests that the enhanced conjugation efficiency is independent of secreted substances. Mutagenesis analysis indicated that the clustered regularly interspaced short palindromic repeats system and the selected restriction-modification systems (Cj0030/Cj0031, Cj0139/Cj0140, Cj0690c, and HsdR) were dispensable for heat shock-enhanced conjugation in C. jejuni. Taking all results together, this study demonstrated a heat shock-enhanced conjugation efficiency in standard C. jejuni strains, leading to an optimized conjugation protocol for molecular manipulation of this organism. The findings from this study also represent a significant step toward elucidation of the molecular mechanism of conjugative gene transfer in C. jejuni. PMID:25911489

  9. Heat Shock-Enhanced Conjugation Efficiency in Standard Campylobacter jejuni Strains.

    PubMed

    Zeng, Ximin; Ardeshna, Devarshi; Lin, Jun

    2015-07-01

    Campylobacter jejuni, the leading bacterial cause of human gastroenteritis in the United States, displays significant strain diversity due to horizontal gene transfer. Conjugation is an important horizontal gene transfer mechanism contributing to the evolution of bacterial pathogenesis and antimicrobial resistance. It has been observed that heat shock could increase transformation efficiency in some bacteria. In this study, the effect of heat shock on C. jejuni conjugation efficiency and the underlying mechanisms were examined. With a modified Escherichia coli donor strain, different C. jejuni recipient strains displayed significant variation in conjugation efficiency ranging from 6.2 × 10(-8) to 6.0 × 10(-3) CFU per recipient cell. Despite reduced viability, heat shock of standard C. jejuni NCTC 11168 and 81-176 strains (e.g., 48 to 54°C for 30 to 60 min) could dramatically enhance C. jejuni conjugation efficiency up to 1,000-fold. The phenotype of the heat shock-enhanced conjugation in C. jejuni recipient cells could be sustained for at least 9 h. Filtered supernatant from the heat shock-treated C. jejuni cells could not enhance conjugation efficiency, which suggests that the enhanced conjugation efficiency is independent of secreted substances. Mutagenesis analysis indicated that the clustered regularly interspaced short palindromic repeats system and the selected restriction-modification systems (Cj0030/Cj0031, Cj0139/Cj0140, Cj0690c, and HsdR) were dispensable for heat shock-enhanced conjugation in C. jejuni. Taking all results together, this study demonstrated a heat shock-enhanced conjugation efficiency in standard C. jejuni strains, leading to an optimized conjugation protocol for molecular manipulation of this organism. The findings from this study also represent a significant step toward elucidation of the molecular mechanism of conjugative gene transfer in C. jejuni. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  10. Campylobacter jejuni type VI secretion system: roles in adaptation to deoxycholic acid, host cell adherence, invasion, and in vivo colonization.

    PubMed

    Lertpiriyapong, Kvin; Gamazon, Eric R; Feng, Yan; Park, Danny S; Pang, Jassia; Botka, Georgina; Graffam, Michelle E; Ge, Zhongming; Fox, James G

    2012-01-01

    The recently identified type VI secretion system (T6SS) of proteobacteria has been shown to promote pathogenicity, competitive advantage over competing microorganisms, and adaptation to environmental perturbation. By detailed phenotypic characterization of loss-of-function mutants, in silico, in vitro and in vivo analyses, we provide evidence that the enteric pathogen, Campylobacter jejuni, possesses a functional T6SS and that the secretion system exerts pleiotropic effects on two crucial processes--survival in a bile salt, deoxycholic acid (DCA), and host cell adherence and invasion. The expression of T6SS during initial exposure to the upper range of physiological levels of DCA (0.075%-0.2%) was detrimental to C. jejuni proliferation, whereas down-regulation or inactivation of T6SS enabled C. jejuni to resist this effect. The C. jejuni multidrug efflux transporter gene, cmeA, was significantly up-regulated during the initial exposure to DCA in the wild type C. jejuni relative to the T6SS-deficient strains, suggesting that inhibition of proliferation is the consequence of T6SS-mediated DCA influx. A sequential modulation of the efflux transporter activity and the T6SS represents, in part, an adaptive mechanism for C. jejuni to overcome this inhibitory effect, thereby ensuring its survival. C. jejuni T6SS plays important roles in host cell adhesion and invasion as T6SS inactivation resulted in a reduction of adherence to and invasion of in vitro cell lines, while over-expression of a hemolysin co-regulated protein, which encodes a secreted T6SS component, greatly enhanced these processes. When inoculated into B6.129P2-IL-10(tm1Cgn) mice, the T6SS-deficient C. jejuni strains did not effectively establish persistent colonization, indicating that T6SS contributes to colonization in vivo. Taken together, our data demonstrate the importance of bacterial T6SS in host cell adhesion, invasion, colonization and, for the first time to our knowledge, adaptation to DCA

  11. The civRT operon is important for Campylobacter jejuni strain 81-176 host cell interactions through regulation of the formate dehydrogenase operon

    USDA-ARS?s Scientific Manuscript database

    C. jejuni colonizes the intestinal mucosa, and the severity of disease in different strains is correlated with host cell interaction and invasion. A microarray screen to identify genes differentially regulated during C. jejuni interaction with tissue culture cells revealed the up-regulation of a two...

  12. Survival of Campylobacter jejuni inoculated into ground beef.

    PubMed Central

    Stern, N J; Kotula, A W

    1982-01-01

    Ground beef was inoculated with mixed cultures of Campylobacter jejuni, and the samples were subjected to various cooking and cold-storage temperatures. When samples were heated in an oven at either 190 or 218 degrees C, approximately 10(7) cells of C. jejuni per g were inactivated (less than 30 cells per g) in less than 10 min after the ground beef reached an internal temperature of 70 degrees C. When the samples were held at -15 degrees C over 14 days of storage, the numbers of C. jejuni declined by 3 log10. When inoculated samples were stored with an equal amount of Cary-Blair diluent at 4 degrees C, no changes in viability were observed over 14 days of storage. Twenty-five times as much C. jejuni was recovered from inoculated ground beef when either 10% glycerol or 10% dimethyl sulfoxide was added to an equal amount of ground beef before freezing as was recovered from peptone-diluted ground beef. Twice as much inoculated C. jejuni was recovered from ground beef plus Cary-Blair diluent as was recovered from ground beef plus peptone diluent. PMID:7181502

  13. Phage-displayed peptides selected for binding to Campylobacter jejuni are antimicrobial.

    PubMed

    Bishop-Hurley, Sharon L; Rea, Philippa J; McSweeney, Christopher S

    2010-10-01

    In developed countries, Campylobacter jejuni is a leading cause of zoonotic bacterial gastroenteritis in humans with chicken meat implicated as a source of infection. Campylobacter jejuni colonises the lower gastrointestinal tract of poultry and during processing is spread from the gastrointestinal tract onto the surface of dressed carcasses. Controlling or eliminating C.jejuni on-farm is considered to be one of the best strategies for reducing human infection. Molecules on the cell surface of C.jejuni interact with the host to facilitate its colonisation and persistence in the gastrointestinal tract of poultry. We used a subtractive phage-display protocol to affinity select for peptides binding to the cell surface of a poultry isolate of C.jejuni with the aim of finding peptides that could be used to control this microorganism in chickens. In total, 27 phage peptides, representing 11 unique clones, were found to inhibit the growth of C.jejuni by up to 99.9% in vitro. One clone was bactericidal, reducing the viability of C.jejuni by 87% in vitro. The phage peptides were highly specific. They completely inhibited the growth of two of the four poultry isolates of C.jejuni tested with no activity detected towards other Gram-negative and Gram-positive bacteria.

  14. Assays to Study the Interaction of Campylobacter jejuni with the Mucosal Surface.

    PubMed

    Clyne, Marguerite; Duggan, Gina; Dunne, Ciara; Dolan, Brendan; Alvarez, Luis; Bourke, Billy

    2017-01-01

    Mucosal colonization and overcoming the mucosal barrier are essential steps in the establishment of infection by Campylobacter jejuni. The interaction between C. jejuni and host cells, including binding and invasion, is thought to be the key virulence factor important for pathogenesis of C. jejuni infections in animals or humans. The intestinal mucosal barrier is composed of a polarized epithelium covered by a thick adherent mucus gel layer. There is a requirement for cell culture assays of infection to accurately represent the in vivo mucosal surface. In this chapter, we describe the use of a number of cell culture models and the use of polarized in vitro organ culture to examine the interaction of C. jejuni with mucosal surfaces.

  15. Common genomic features of Campylobacter jejuni subsp. doylei strains distinguish them from C. jejuni subsp. jejuni

    PubMed Central

    Parker, Craig T; Miller, William G; Horn, Sharon T; Lastovica, Albert J

    2007-01-01

    Background Campylobacter jejuni has been divided into two subspecies: C. jejuni subsp. jejuni (Cjj) and C. jejuni subsp. doylei (Cjd). Nearly all of the C. jejuni strains isolated are Cjj; nevertheless, although Cjd strains are isolated infrequently, they differ from Cjj in two key aspects: they are obtained primarily from human clinical samples and are associated often with bacteremia, in addition to gastroenteritis. In this study, we utilized multilocus sequence typing (MLST) and a DNA microarray-based comparative genomic indexing (CGI) approach to examine the genomic diversity and gene content of Cjd strains. Results A geographically diverse collection of eight Cjd strains was examined by MLST and determined to be phylogenetically distinct from Cjj strains. Microarray-based CGI approach also supported this. We were able to demonstrate that Cjd strains exhibited divergence from Cjj strains NCTC 11168 and RM1221 in many of the intraspecies hypervariable regions. Moreover, multiple metabolic, transport and virulence functions (e.g. cytolethal distending toxin) were shown to be absent in the Cjd strains examined. Conclusion Our data demonstrate that Cjd are phylogenetically distinct from Cjj strains. Using the CGI approach, we identified subsets of absent genes from amongst the C. jejuni genes that provide clues as to the potential evolutionary origin and unusual pathogenicity of Cjd. PMID:17535437

  16. Chicken Juice Enhances Surface Attachment and Biofilm Formation of Campylobacter jejuni

    PubMed Central

    Brown, Helen L.; Reuter, Mark; Salt, Louise J.; Cross, Kathryn L.; Betts, Roy P.

    2014-01-01

    The bacterial pathogen Campylobacter jejuni is primarily transmitted via the consumption of contaminated foodstuffs, especially poultry meat. In food processing environments, C. jejuni is required to survive a multitude of stresses and requires the use of specific survival mechanisms, such as biofilms. An initial step in biofilm formation is bacterial attachment to a surface. Here, we investigated the effects of a chicken meat exudate (chicken juice) on C. jejuni surface attachment and biofilm formation. Supplementation of brucella broth with ≥5% chicken juice resulted in increased biofilm formation on glass, polystyrene, and stainless steel surfaces with four C. jejuni isolates and one C. coli isolate in both microaerobic and aerobic conditions. When incubated with chicken juice, C. jejuni was both able to grow and form biofilms in static cultures in aerobic conditions. Electron microscopy showed that C. jejuni cells were associated with chicken juice particulates attached to the abiotic surface rather than the surface itself. This suggests that chicken juice contributes to C. jejuni biofilm formation by covering and conditioning the abiotic surface and is a source of nutrients. Chicken juice was able to complement the reduction in biofilm formation of an aflagellated mutant of C. jejuni, indicating that chicken juice may support food chain transmission of isolates with lowered motility. We provide here a useful model for studying the interaction of C. jejuni biofilms in food chain-relevant conditions and also show a possible mechanism for C. jejuni cell attachment and biofilm initiation on abiotic surfaces within the food chain. PMID:25192991

  17. Hyperosmotic Stress Response of Campylobacter jejuni

    PubMed Central

    Cameron, Andrew; Frirdich, Emilisa; Huynh, Steven; Parker, Craig T.

    2012-01-01

    The diarrheal pathogen Campylobacter jejuni and other gastrointestinal bacteria encounter changes in osmolarity in the environment, through exposure to food processing, and upon entering host organisms, where osmotic adaptation can be associated with virulence. In this study, growth profiles, transcriptomics, and phenotypic, mutant, and single-cell analyses were used to explore the effects of hyperosmotic stress exposure on C. jejuni. Increased growth inhibition correlated with increased osmotic concentration, with both ionic and nonionic stressors inhibiting growth at 0.620 total osmol liter−1. C. jejuni adaptation to a range of osmotic stressors and concentrations was accompanied by severe filamentation in subpopulations, with microscopy indicating septum formation and phenotypic diversity between individual cells in a filament. Population heterogeneity was also exemplified by the bifurcation of colony morphology into small and large variants on salt stress plates. Flow cytometry of C. jejuni harboring green fluorescent protein (GFP) fused to the ATP synthase promoter likewise revealed bimodal subpopulations under hyperosmotic stress. We also identified frequent hyperosmotic stress-sensitive variants within the clonal wild-type population propagated on standard laboratory medium. Microarray analysis following hyperosmotic upshift revealed enhanced expression of heat shock genes and genes encoding enzymes for synthesis of potential osmoprotectants and cross-protective induction of oxidative stress genes. The capsule export gene kpsM was also upregulated, and an acapsular mutant was defective for growth under hyperosmotic stress. For C. jejuni, an organism lacking most conventional osmotic response factors, these data suggest an unusual hyperosmotic stress response, including likely “bet-hedging” survival strategies relying on the presence of stress-fit individuals in a heterogeneous population. PMID:22961853

  18. Alpinia katsumadai Extracts Inhibit Adhesion and Invasion of Campylobacter jejuni in Animal and Human Foetal Small Intestine Cell Lines.

    PubMed

    Pogačar, Maja Šikić; Klančnik, Anja; Bucar, Franz; Langerholc, Tomaž; Možina, Sonja Smole

    2015-10-01

    Alpinia katsumadai is used in traditional Chinese medicine for abdominal distention, pain, and diarrhoea. Campylobacter jejuni is the most common cause of bacterial food-borne diarrhoeal illnesses worldwide. Adhesion to gut epithelium is a prerequisite in its pathogenesis. The antimicrobial, cytotoxic, and anti-adhesive activities of a chemically characterised extract (SEE) and its residual material of hydrodistillation (hdSEE-R) from A. katsumadai seeds were evaluated against C. jejuni. Minimal inhibitory concentrations for SEE and hdSEE-R were 0.5 mg/mL and 0.25 mg/mL, respectively, and there was no cytotoxic influence in the anti-adhesion tests, as these were performed at much lower concentrations of these tested plant extracts. Adhesion of C. jejuni to pig (PSI) and human foetal (H4) small-intestine cell lines was significantly decreased at lower concentrations (0.2 to 50 µg/mL). In the same concentration range, the invasiveness of C. jejuni in PSI cells was reduced by 45% to 65% when they were treated with SEE or hdSEE-R. The hdSEE-R represents a bioactive waste with a high phenolic content and an anti-adhesive activity against C. jejuni and thus has the potential for use in pharmaceutical and food products. Copyright © 2015 John Wiley & Sons, Ltd.

  19. Mutation in the peb1A Locus of Campylobacter jejuni Reduces Interactions with Epithelial Cells and Intestinal Colonization of Mice

    PubMed Central

    Pei, Zhiheng; Burucoa, Christophe; Grignon, Bernadette; Baqar, Shahida; Huang, Xiao-Zhe; Kopecko, Dennis J.; Bourgeois, A. L.; Fauchere, Jean-Louis; Blaser, Martin J.

    1998-01-01

    Campylobacter jejuni is one of the leading causes of bacterial diarrhea throughout the world. We previously found that PEB1 is a homolog of cluster 3 binding proteins of bacterial ABC transporters and that a C. jejuni adhesin, cell-binding factor 1 (CBF1), if not identical to, contains PEB1. A single protein migrating at approximately 27 to 28 kDa was recognized by anti-CBF1 and anti-PEB1. To determine the role that the operon encoding PEB1 plays in C. jejuni adherence, peb1A, the gene encoding PEB1, was disrupted in strain 81-176 by insertion of a kanamycin resistance gene through homologous recombination. Inactivation of this operon completely abolished expression of CBF1, as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting. In comparison to the wild-type strain, the mutant strain showed 50- to 100-fold less adherence to and 15-fold less invasion of epithelial cells in culture. Mouse challenge studies showed that the rate and duration of intestinal colonization by the mutant were significantly lower and shorter than with the wild-type strain. In summary, PEB1 is identical to a previously identified cell-binding factor, CBF1, in C. jejuni, and the peb1A locus plays an important role in epithelial cell interactions and in intestinal colonization in a mouse model. PMID:9488379

  20. Autoinducer-2 Production in Campylobacter jejuni Contributes to Chicken Colonization ▿

    PubMed Central

    Quiñones, Beatriz; Miller, William G.; Bates, Anna H.; Mandrell, Robert E.

    2009-01-01

    Inactivation of luxS, encoding an AI-2 biosynthesis enzyme, in Campylobacter jejuni strain 81-176 significantly reduced colonization of the chick lower gastrointestinal tract, chemotaxis toward organic acids, and in vitro adherence to LMH chicken hepatoma cells. Thus, AI-2 production in C. jejuni contributes to host colonization and interactions with epithelial cells. PMID:19011073

  1. N-Glycosylation of Campylobacter jejuni Surface Proteins Promotes Bacterial Fitness

    PubMed Central

    Nothaft, Harald; Zheng, Jing

    2013-01-01

    Campylobacter jejuni is the etiologic agent of human bacterial gastroenteritis worldwide. In contrast, despite heavy colonization, C. jejuni maintains a commensal mode of existence in chickens. The consumption of contaminated chicken products is thought to be the principal mode of C. jejuni transmission to the human population. C. jejuni harbors a system for N-linked protein glycosylation that has been well characterized and modifies more than 60 periplasmic and membrane-bound proteins. However, the precise role of this modification in the biology of C. jejuni remains unexplored. We hypothesized that the N-glycans protect C. jejuni surface proteins from the action of gut proteases. The C. jejuni pglB mutant, deficient in the expression of the oligosaccharyltransferase, exhibited reduced growth in medium supplemented with chicken cecal contents (CCC) compared with that of wild-type (WT) cells. Inactivation of the cecal proteases by heat treatment or with protease inhibitors completely restored bacterial viability and partially rescued bacterial growth. Physiological concentrations of trypsin, but not chymotrypsin, also reduced C. jejuni pglB mutant CFU. Live or dead staining indicated that CCC preferentially influenced C. jejuni growth as opposed to bacterial viability. We identified multiple chicken cecal proteases by mass fingerprinting. The use of protease inhibitors that target specific classes indicated that both metalloproteases and serine proteases were involved in the attenuated growth of the oligosaccharyltransferase mutant. In conclusion, protein N-linked glycosylation of surface proteins may enhance C. jejuni fitness by protecting bacterial proteins from cleavage due to gut proteases. PMID:23460522

  2. Occurrence of the invasion associated marker (iam) in Campylobacter jejuni isolated from cattle

    PubMed Central

    2011-01-01

    Background The invasion associated marker (iam) has been detected in the majority of invasive Campylobacter jejuni retrieved from humans. Furthermore, the detection of iam in C. jejuni isolated from two important hosts, humans and chickens, suggested a role for this marker in C. jejuni's colonization of multiple hosts. However, no data exist regarding the occurrence of this marker in C. jejuni isolated from non-poultry food-animals such as cattle, an increasingly important source for human infections. Since little is known about the genetics associated with C. jejuni's capability for colonizing physiologically disparate hosts, we investigated the occurrence of the iam in C. jejuni isolated from cattle and assessed the potential of iam-containing cattle and human isolates for chicken colonization and human cell invasion. Results Simultaneous RAPD typing and iam-specific PCR analysis of 129 C. jejuni isolated from 1171 cattle fecal samples showed that 8 (6.2%) of the isolates were iam-positive, while 7 (54%) of human-associated isolates were iam-positive. The iam sequences were mostly heterogeneous and occurred in diverse genetic backgrounds. All iam-positive isolates were motile and possessed important genes (cadF, ciaB, cdtB) associated with adhesion and virulence. Although certain iam-containing isolates invaded and survived in INT-407 cells in high numbers and successfully colonized live chickens, there was no clear association between the occurrence, allelic sequence, and expression levels of the iam and the aforementioned phenotypes. Conclusions We show that the prevalence of iam in cattle C. jejuni is relatively lower as compared to isolates occurring in humans and chickens. In addition, iam was polymorphic and certain alleles occur in cattle isolates that were capable of colonizing and invading chickens and human intestinal cells, respectively. However, the iam did not appear to contribute to the cattle-associated C. jejuni's potential for invasion and

  3. Natural Transformation of Campylobacter jejuni Occurs Beyond Limits of Growth

    PubMed Central

    Vegge, Christina S.; Brøndsted, Lone; Ligowska-Marzęta, Małgorzata; Ingmer, Hanne

    2012-01-01

    Campylobacter jejuni is a human bacterial pathogen. While poultry is considered to be a major source of food borne campylobacteriosis, C. jejuni is frequently found in the external environment, and water is another well-known source of human infections. Natural transformation is considered to be one of the main mechanisms for mediating transfer of genetic material and evolution of the organism. Given the diverse habitats of C. jejuni we set out to examine how environmental conditions and physiological processes affect natural transformation of C. jejuni. We show that the efficiency of transformation is correlated to the growth conditions, but more importantly that transformation occurs at growth-restrictive conditions as well as in the late stationary phase; hence revealing that growth per se is not required for C. jejuni to be competent. Yet, natural transformation of C. jejuni is an energy dependent process, that occurs in the absence of transcription but requires an active translational machinery. Moreover, we show the ATP dependent ClpP protease to be important for transformation, which possibly could be associated with reduced protein glycosylation in the ClpP mutant. In contrast, competence of C. jejuni was neither found to be involved in DNA repair following DNA damage nor to provide a growth benefit. Kinetic studies revealed that several transformation events occur per cell cycle indicating that natural transformation of C. jejuni is a highly efficient process. Thus, our findings suggest that horizontal gene transfer by natural transformation takes place in various habitats occupied by C. jejuni. PMID:23049803

  4. Enhanced transmission of antibiotic resistance in Campylobacter jejuni biofilms by natural transformation.

    PubMed

    Bae, Junghee; Oh, Euna; Jeon, Byeonghwa

    2014-12-01

    Campylobacter jejuni is a leading food-borne pathogen, and its antibiotic resistance is of serious concern to public health worldwide. C. jejuni is naturally competent for DNA transformation and freely takes up foreign DNA harboring genetic information responsible for antibiotic resistance. In this study, we demonstrate that C. jejuni transfers antibiotic resistance genes more frequently in biofilms than in planktonic cells by natural transformation. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

  5. Relaxation of DNA supercoiling leads to increased invasion of epithelial cells and protein secretion by Campylobacter jejuni

    USDA-ARS?s Scientific Manuscript database

    Invasion of intestinal epithelial cells by Campylobacter jejuni is a critical step during infection of the human intestine by this important human pathogen. In this study we investigated the role played by DNA supercoiling in the regulation of invasion of epithelial cells and the mechanism by which ...

  6. Antimicrobial effect of electrolyzed water for inactivating Campylobacter jejuni during poultry washing.

    PubMed

    Park, Hoon; Hung, Yen-Con; Brackett, Robert E

    2002-01-30

    The effectiveness of electrolyzed (EO) water for killing Campylobacter jejuni on poultry was evaluated. Complete inactivation of C. jejuni in pure culture occurred within 10 s after exposure to EO or chlorinated water, both of which contained 50 mg/l of residual chlorine. A strong bactericidal activity was also observed on the diluted EO water (containing 25 mg/l of residual chlorine) and the mean population of C. jejuni was reduced to less than 10 CFU/ml (detected only by enrichment for 48 h) after 10-s treatment. The diluted chlorine water (25 mg/l residual chlorine) was less effective than the diluted EO water for inactivation of C. jejuni. EO water was further evaluated for its effectiveness in reducing C. jejuni on chicken during washing. EO water treatment was equally effective as chlorinated water and both achieved reduction of C. jejuni by about 3 log10 CFU/g on chicken, whereas deionized water (control) treatment resulted in only 1 log10 CFU/g reduction. No viable cells of C. jejuni were recovered in EO and chlorinated water after washing treatment, whereas high populations of C. jejuni (4 log10 CFU/ml) were recovered in the wash solution after the control treatment. Our study demonstrated that EO water was very effective not only in reducing the populations of C. jejuni on chicken, but also could prevent cross-contamination of processing environments.

  7. Methylation-dependent DNA discrimination in natural transformation of Campylobacter jejuni

    PubMed Central

    Leveque, Rhiannon M.; Dawid, Suzanne; DiRita, Victor J.

    2017-01-01

    Campylobacter jejuni, a leading cause of bacterial gastroenteritis, is naturally competent. Like many competent organisms, C. jejuni restricts the DNA that can be used for transformation to minimize undesirable changes in the chromosome. Although C. jejuni can be transformed by C. jejuni-derived DNA, it is poorly transformed by the same DNA propagated in Escherichia coli or produced with PCR. Our work indicates that methylation plays an important role in marking DNA for transformation. We have identified a highly conserved DNA methyltransferase, which we term Campylobacter transformation system methyltransferase (ctsM), which methylates an overrepresented 6-bp sequence in the chromosome. DNA derived from a ctsM mutant transforms C. jejuni significantly less well than DNA derived from ctsM+ (parental) cells. The ctsM mutation itself does not affect transformation efficiency when parental DNA is used, suggesting that CtsM is important for marking transforming DNA, but not for transformation itself. The mutant has no growth defect, arguing against ongoing restriction of its own DNA. We further show that E. coli plasmid and PCR-derived DNA can efficiently transform C. jejuni when only a subset of the CtsM sites are methylated in vitro. A single methylation event 1 kb upstream of the DNA involved in homologous recombination is sufficient to transform C. jejuni, whereas otherwise identical unmethylated DNA is not. Methylation influences DNA uptake, with a slight effect also seen on DNA binding. This mechanism of DNA discrimination in C. jejuni is distinct from the DNA discrimination described in other competent bacteria. PMID:28855338

  8. Fate of Campylobacter jejuni in butter.

    PubMed

    Zhao, T; Doyle, M P; Berg, D E

    2000-01-01

    An outbreak of Campylobacter enteritis was associated with a restaurant in Louisiana during the summer of 1995. Thirty cases were identified, and four required hospitalization. Campylobacter jejuni was isolated from the patients, and epidemiologic studies revealed illness associated with eating garlic butter served at the restaurant. Three batches of garlic butter prepared by the restaurant associated with the outbreak and a C. jejuni isolate obtained from a patient involved in the outbreak were used for studies to determine the fate of C. jejuni in garlic butter. Studies also were done to determine the efficacy of the heat treatment used by the restaurant to prepare garlic bread to kill C. jejuni. Garlic butter was inoculated with approximately 10(4) and 10(6) CFU/g of C. jejuni and held at 5 or 21 degrees C. Results revealed that the survival of C. jejuni differed greatly, depending on the presence or absence of garlic. At 5 degrees C, C. jejuni populations decreased to an undetectable level (<10 CFU/g) within 3 h for two batches and within 24 h for another batch. In contrast, C. jejuni could survive at 5 degrees C for 13 days in butter with no garlic. At 21 degrees C, C. jejuni populations decreased to an undetectable level within 5 h for two batches and to 50 CFU/g in 5 h for another batch. In contrast, C. jejuni was detected at 500 CFU/g at 28 h after inoculation but was undetectable at 3 days in butter with no garlic held at 21 degrees C. The heating procedure (135 degrees C, 4 min) used to make garlic bread by the implicated restaurant was determined not to be sufficient for killing C. jejuni, with the internal temperature of the buttered bread after heating ranging from 19 to 22 degrees C. This study revealed that C. jejuni can survive for many days in refrigerated butter, but large populations (10(3) to 10(5) CFU/g) are killed within a few hours in butter that contains garlic. Furthermore, the heat treatment used by the restaurant to melt garlic butter in

  9. Anti-Campylobacter activity of resveratrol and an extract from waste Pinot noir grape skins and seeds, and resistance of Camp. jejuni planktonic and biofilm cells, mediated via the CmeABC efflux pump.

    PubMed

    Klančnik, A; Šikić Pogačar, M; Trošt, K; Tušek Žnidarič, M; Mozetič Vodopivec, B; Smole Možina, S

    2017-01-01

    To define anti-Campylobacter jejuni activity of an extract from waste skins and seeds of Pinot noir grapes (GSS), resveratrol and possible resistance mechanisms, and the influence of these on Camp. jejuni morphology. Using gene-specific knock-out Camp. jejuni mutants and an efflux pump inhibitor, we showed CmeABC as the most active efflux pump for extrusion across the outer membrane of GSS extract and resveratrol. Using polystyrene surface and pig small intestine epithelial (PSI) and human foetal small intestine (H4) cell lines, GSS extract shows an efficient inhibition of adhesion of Camp. jejuni to these abiotic and biotic surfaces. Low doses of GSS extract can inhibit Camp. jejuni adhesion to polystyrene surfaces and to PSI and H4 cells, and can thus modulate Camp. jejuni invasion and intracellular survival. An understanding of the activities of GSS extract and resveratrol as bacterial growth inhibitors and the specific mechanisms of cell accumulation is crucial for our understanding of Camp. jejuni resistance. GSS extract inhibition of Camp. jejuni adhesion to abiotic and biotic surfaces provides a further step towards the application of new innovative strategies to control Campylobacter contamination and infection via the food chain. © 2016 The Society for Applied Microbiology.

  10. Strain-dependent induction of epithelial cell oncosis by Campylobacter jejuni is correlated with invasion ability and is independent of cytolethal distending toxin.

    PubMed

    Kalischuk, Lisa D; Inglis, G Douglas; Buret, Andre G

    2007-09-01

    Induction of host cell death is thought to play an important role in bacterial pathogenesis. Campylobacter jejuni is a prevalent cause of bacterial enteritis; however, its effects on enterocytes remain unclear. The present study indicates for the first time that C. jejuni induces oncotic, rather than apoptotic death of T84 enterocytes. C. jejuni-treated enterocytes exhibited extensive cytoplasmic vacuolation, rapid (3-6 h) loss of plasma membrane integrity ('cytotoxicity'), loss of mitochondrial transmembrane potential, and ATP depletion. Enterocytes also exhibited increased oligonucleosomal DNA fragmentation, a feature characteristic of apoptosis. However, consistent with a non-apoptotic process, DNA fragmentation and cytotoxicity were not caspase dependent. During apoptosis, caspases mediate cleavage of poly(ADP-ribose) polymerase; however, cleavage was not observed in C. jejuni-treated monolayers. Cytotoxicity, ATP depletion and DNA fragmentation were not prevented by the deletion of the cytolethal distending toxin (CDT) gene, indicating that C. jejuni causes enterocyte oncosis via a mechanism that is CDT independent. The ability to cause oncosis was significantly decreased in a FlaAFlaB mutant (CDT(+)) that was defective in the ability to adhere and invade enterocytes. Analysis of clinical isolates revealed that oncosis was strain dependent and correlated with increased invasive ability. These observations offer new insights into the pathogenesis of C. jejuni infection.

  11. Accumulation of Peptidoglycan O-Acetylation Leads to Altered Cell Wall Biochemistry and Negatively Impacts Pathogenesis Factors of Campylobacter jejuni.

    PubMed

    Ha, Reuben; Frirdich, Emilisa; Sychantha, David; Biboy, Jacob; Taveirne, Michael E; Johnson, Jeremiah G; DiRita, Victor J; Vollmer, Waldemar; Clarke, Anthony J; Gaynor, Erin C

    2016-10-21

    Campylobacter jejuni is a leading cause of bacterial gastroenteritis in the developed world. Despite its prevalence, its mechanisms of pathogenesis are poorly understood. Peptidoglycan (PG) is important for helical shape, colonization, and host-pathogen interactions in C. jejuni Therefore, changes in PG greatly impact the physiology of this organism. O-acetylation of peptidoglycan (OAP) is a bacterial phenomenon proposed to be important for proper cell growth, characterized by acetylation of the C6 hydroxyl group of N-acetylmuramic acid in the PG glycan backbone. The OAP gene cluster consists of a PG O-acetyltransferase A (patA) for translocation of acetate into the periplasm, a PG O-acetyltransferase B (patB) for O-acetylation, and an O-acetylpeptidoglycan esterase (ape1) for de-O-acetylation. In this study, reduced OAP in ΔpatA and ΔpatB had minimal impact on C. jejuni growth and fitness under the conditions tested. However, accumulation of OAP in Δape1 resulted in marked differences in PG biochemistry, including O-acetylation, anhydromuropeptide levels, and changes not expected to result directly from Ape1 activity. This suggests that OAP may be a form of substrate level regulation in PG biosynthesis. Ape1 acetylesterase activity was confirmed in vitro using p-nitrophenyl acetate and O-acetylated PG as substrates. In addition, Δape1 exhibited defects in pathogenesis-associated phenotypes, including cell shape, motility, biofilm formation, cell surface hydrophobicity, and sodium deoxycholate sensitivity. Δape1 was also impaired for chick colonization and adhesion, invasion, intracellular survival, and induction of IL-8 production in INT407 cells in vitro The importance of Ape1 in C. jejuni biology makes it a good candidate as an antimicrobial target. © 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

  12. Inactivation of Campylobacter jejuni by chlorine and monochloramine.

    PubMed Central

    Blaser, M J; Smith, P F; Wang, W L; Hoff, J C

    1986-01-01

    Campylobacter jejuni and closely related organisms are important bacterial causes of acute diarrheal illness in the United States. Both endemic and epidemic infections have been associated with consuming untreated or improperly treated surface water. We compared susceptibility of three C. jejuni strains and Escherichia coli ATCC 11229 with standard procedures used to disinfect water. Inactivation of bacterial preparations with 0.1 mg of chlorine and 1.0 mg of monochloramine per liter was determined at pH 6 and 8 and at 4 and 25 degrees C. Under virtually every condition tested, each of the three C. jejuni strains was more susceptible than the E. coli control strain, with greater than 99% inactivation after 15 min of contact with 1.0 mg of monochloramine per liter or 5 min of contact with 0.1 mg of free chlorine per liter. Results of experiments in which an antibiotic-containing medium was used suggest that a high proportion of the remaining cells were injured. An animal-passaged C. jejuni strain was as susceptible to chlorine disinfection as were laboratory-passaged strains. These results suggest that disinfection procedures commonly used for treatment of drinking water to remove coliform bacteria are adequate to eliminate C. jejuni and further correlate with the absence of outbreaks associated with properly treated water. PMID:3954344

  13. Microfluidics meets metabolomics to reveal the impact of Campylobacter jejuni infection on biochemical pathways.

    PubMed

    Mortensen, Ninell P; Mercier, Kelly A; McRitchie, Susan; Cavallo, Tammy B; Pathmasiri, Wimal; Stewart, Delisha; Sumner, Susan J

    2016-06-01

    Microfluidic devices that are currently being used in pharmaceutical research also have a significant potential for utilization in investigating exposure to infectious agents. We have established a microfluidic device cultured with Caco-2 cells, and utilized metabolomics to investigate the biochemical responses to the bacterial pathogen Campylobacter jejuni. In the microfluidic devices, Caco-2 cells polarize at day 5, are uniform, have defined brush borders and tight junctions, and form a mucus layer. Metabolomics analysis of cell culture media collected from both Caco-2 cell culture systems demonstrated a more metabolic homogenous biochemical profile in the media collected from microfluidic devices, compared with media collected from transwells. GeneGo pathway mapping indicated that aminoacyl-tRNA biosynthesis was perturbed by fluid flow, suggesting that fluid dynamics and shear stress impacts the cells translational quality control. Both microfluidic device and transwell culturing systems were used to investigate the impact of Campylobacter jejuni infection on biochemical processes. Caco-2 cells cultured in either system were infected at day 5 with C. jejuni 81-176 for 48 h. Metabolomics analysis clearly differentiated C. jejuni 81-176 infected and non-infected medias collected from the microfluidic devices, and demonstrated that C. jejuni 81-176 infection in microfluidic devices impacts branched-chain amino acid metabolism, glycolysis, and gluconeogenesis. In contrast, no distinction was seen in the biochemical profiles of infected versus non-infected media collected from cells cultured in transwells. Microfluidic culturing conditions demonstrated a more metabolically homogenous cell population, and present the opportunity for studying host-pathogen interactions for extended periods of time.

  14. Microfluidics Meets Metabolomics to Reveal the Impact of Campylobacter jejuni Infection on Biochemical Pathways

    PubMed Central

    Mortensen, Ninell P.; Mercier, Kelly A.; McRitchie, Susan; Cavallo, Tammy B.; Pathmasiri, Wimal; Stewart, Delisha; Sumner, Susan J.

    2016-01-01

    Microfluidic devices that are currently being used in pharmaceutical research also have a significant potential for utilization in investigating exposure to infectious agents. We have established a microfluidic device cultured with Caco-2 cells, and utilized metabolomics to investigate the biochemical responses to the bacterial pathogen Campylobacter jejuni. In the microfluidic devices, Caco-2 cells polarize at day 5, are uniform, have defined brush borders and tight junctions, and form a mucus layer. Metabolomics analysis of cell culture media collected from both Caco-2 cell culture systems demonstrated a more metabolic homogenous biochemical profile in the media collected from microfluidic devices, compared with media collected from transwells. GeneGo pathway mapping indicated that aminoacyl-tRNA biosynthesis was perturbed by fluid flow, suggesting that fluid dynamics and shear stress impacts the cells translational quality control. Both microfluidic device and transwell culturing systems were used to investigate the impact of Campylobacter jejuni infection on biochemical processes. Caco-2 cells cultured in either system were infected at day 5 with C. jejuni 81-176 for 48 hours. Metabolomics analysis clearly differentiated C. jejuni 81-176 infected and non-infected medias collected from the microfluidic devices, and demonstrated that C. jejuni 81-176 infection in microfluidic devices impacts branched-chain amino acid metabolism, glycolysis, and gluconeogenesis. In contrast, no distinction was seen in the biochemical profiles of infected versus non-infected media collected from cells cultured in transwells. Microfluidic culturing conditions demonstrated a more metabolically homogenous cell population, and present the opportunity for studying host-pathogen interactions for extended periods of time. PMID:27231016

  15. Cytolethal Distending Toxin From Campylobacter jejuni Requires the Cytoskeleton for Toxic Activity.

    PubMed

    Méndez-Olvera, Estela T; Bustos-Martínez, Jaime A; López-Vidal, Yolanda; Verdugo-Rodríguez, Antonio; Martínez-Gómez, Daniel

    2016-10-01

    Campylobacter jejuni is one of the major causes of infectious diarrhea worldwide. The distending cytolethal toxin (CDT) of Campylobacter spp. interferes with normal cell cycle progression. This toxic effect is considered a result of DNase activity that produces chromosomal DNA damage. To perform this event, the toxin must be endocytosed and translocated to the nucleus. The aim of this study was to evaluate the role of the cytoskeleton in the translocation of CDT to the nucleus. Campylobacter jejuni ATCC 33291 and seven isolates donated from Instituto de Biotecnologia were used in this study. The presence of CDT genes in C. jejuni strains was determined by PCR. To evaluate the effect of CDT, HeLa cells were treated with bacterial lysate, and the damage and morphological changes were analyzed by microscopy, immunofluorescence staining, and flow cytometry. To evaluate the role of the cytoskeleton, HeLa cells were treated with either latrunculin A or by nocodazole and analyzed by microscopy, flow cytometry, and immunoquantification (ELISA). The results obtained showed that the eight strains of C. jejuni , including the reference strain, had the ability to produce the toxin. Usage of latrunculin A and nocodazole, two cytoskeletal inhibitors, blocked the toxic effect in cells treated with the toxin. This phenomenon was evident in flow cytometry analysis and immunoquantification of Cdc2-phosphorylated. This work showed that the cytotoxic activity of the C. jejuni CDT is dependent on its endocytosis. The alteration in the microtubules and actin filaments caused a blockage transit of the toxin, preventing it from reaching the nucleus of the cell, as well as preventing DNA fragmentation and alteration of the cell cycle. The CDT toxin appears to be an important element for the pathogenesis of campylobacteriosis, since all clinical isolates showed the presence of cdtA , cdtB and cdtC genes.

  16. Cytolethal Distending Toxin From Campylobacter jejuni Requires the Cytoskeleton for Toxic Activity

    PubMed Central

    Méndez-Olvera, Estela T.; Bustos-Martínez, Jaime A.; López-Vidal, Yolanda; Verdugo-Rodríguez, Antonio; Martínez-Gómez, Daniel

    2016-01-01

    Background Campylobacter jejuni is one of the major causes of infectious diarrhea worldwide. The distending cytolethal toxin (CDT) of Campylobacter spp. interferes with normal cell cycle progression. This toxic effect is considered a result of DNase activity that produces chromosomal DNA damage. To perform this event, the toxin must be endocytosed and translocated to the nucleus. Objectives The aim of this study was to evaluate the role of the cytoskeleton in the translocation of CDT to the nucleus. Methods Campylobacter jejuni ATCC 33291 and seven isolates donated from Instituto de Biotecnologia were used in this study. The presence of CDT genes in C. jejuni strains was determined by PCR. To evaluate the effect of CDT, HeLa cells were treated with bacterial lysate, and the damage and morphological changes were analyzed by microscopy, immunofluorescence staining, and flow cytometry. To evaluate the role of the cytoskeleton, HeLa cells were treated with either latrunculin A or by nocodazole and analyzed by microscopy, flow cytometry, and immunoquantification (ELISA). Results The results obtained showed that the eight strains of C. jejuni, including the reference strain, had the ability to produce the toxin. Usage of latrunculin A and nocodazole, two cytoskeletal inhibitors, blocked the toxic effect in cells treated with the toxin. This phenomenon was evident in flow cytometry analysis and immunoquantification of Cdc2-phosphorylated. Conclusions This work showed that the cytotoxic activity of the C. jejuni CDT is dependent on its endocytosis. The alteration in the microtubules and actin filaments caused a blockage transit of the toxin, preventing it from reaching the nucleus of the cell, as well as preventing DNA fragmentation and alteration of the cell cycle. The CDT toxin appears to be an important element for the pathogenesis of campylobacteriosis, since all clinical isolates showed the presence of cdtA, cdtB and cdtC genes. PMID:27942359

  17. Role of Campylobacter jejuni Infection in the Pathogenesis of Guillain-Barré Syndrome: An Update

    PubMed Central

    Nyati, Kishan Kumar; Nyati, Roopanshi

    2013-01-01

    Our current knowledge on Campylobacter jejuni infections in humans has progressively increased over the past few decades. Infection with C. jejuni is the most common cause of bacterial gastroenteritis, sometimes surpassing other infections due to Salmonella, Shigella, and Escherichia coli. Most infections are acquired due to consumption of raw or undercooked poultry, unpasteurized milk, and contaminated water. After developing the diagnostic methods to detect C. jejuni, the possibility to identify the association of its infection with new diseases has been increased. After the successful isolation of C. jejuni, reports have been published citing the occurrence of GBS following C. jejuni infection. Thus, C. jejuni is now considered as a major triggering agent of GBS. Molecular mimicry between sialylated lipooligosaccharide structures on the cell envelope of these bacteria and ganglioside epitopes on the human nerves that generates cross-reactive immune response results in autoimmune-driven nerve damage. Though C. jejuni is associated with several pathologic forms of GBS, axonal subtypes following C. jejuni infection may be more severe. Ample amount of existing data covers a large spectrum of GBS; however, the studies on C. jejuni-associated GBS are still inconclusive. Therefore, this review provides an update on the C. jejuni infections engaged in the pathogenesis of GBS. PMID:24000328

  18. Virulence characterization of Campylobacter jejuni isolated from resident wild birds in Tokachi area, Japan.

    PubMed

    Shyaka, Anselme; Kusumoto, Akiko; Chaisowwong, Warangkhana; Okouchi, Yoshiki; Fukumoto, Shinya; Yoshimura, Aya; Kawamoto, Keiko

    2015-08-01

    The prevalence of Campylobacter jejuni in wild birds is a potential hazard for human and animal health. The aim of this study was to establish the prevalence of C. jejuni in wild birds in Tokachi area, Hokkaido, Japan and investigate their virulence in vitro. In total, 173 cloacal swabs from individual wild birds were collected for the detection of Campylobacter spp. Thirty four samples (19.7%) were positive for Campylobacter of which 94.1% (32/34 samples) were C. jejuni. Additionally, one C. coli and one C. fetus were isolated. Seven C. jejuni isolates (one from crows and the other from pigeons) had important virulence genes including all three CDT genes (cdtA, cdtB and cdtC) and flaA, flaB, ciaB and cadF, and the other isolates were lacking cdtA gene. Further studies on in vitro virulence-associated phenotypes, such as motility assay on soft agar and invasion assay in Caco-2 cells, were performed. The wild bird C. jejuni isolates adhered and invaded human cells. Although the numbers of viable intracellular bacteria of wild bird isolates were lower than a type strain NCTC11168, they persisted at 48-hr and underwent replication in host cells.

  19. Virulence characterization of Campylobacter jejuni isolated from resident wild birds in Tokachi area, Japan

    PubMed Central

    SHYAKA, Anselme; KUSUMOTO, Akiko; CHAISOWWONG, Warangkhana; OKOUCHI, Yoshiki; FUKUMOTO, Shinya; YOSHIMURA, Aya; KAWAMOTO, Keiko

    2015-01-01

    The prevalence of Campylobacter jejuni in wild birds is a potential hazard for human and animal health. The aim of this study was to establish the prevalence of C. jejuni in wild birds in Tokachi area, Hokkaido, Japan and investigate their virulence in vitro. In total, 173 cloacal swabs from individual wild birds were collected for the detection of Campylobacter spp. Thirty four samples (19.7%) were positive for Campylobacter of which 94.1% (32/34 samples) were C. jejuni. Additionally, one C. coli and one C. fetus were isolated. Seven C. jejuni isolates (one from crows and the other from pigeons) had important virulence genes including all three CDT genes (cdtA, cdtB and cdtC) and flaA, flaB, ciaB and cadF, and the other isolates were lacking cdtA gene. Further studies on in vitro virulence-associated phenotypes, such as motility assay on soft agar and invasion assay in Caco-2 cells, were performed. The wild bird C. jejuni isolates adhered and invaded human cells. Although the numbers of viable intracellular bacteria of wild bird isolates were lower than a type strain NCTC11168, they persisted at 48-hr and underwent replication in host cells. PMID:25843040

  20. High pressure inactivation of Escherichia coli, Campylobacter jejuni, and spoilage microbiota on poultry meat.

    PubMed

    Liu, Yang; Betti, Mirko; Gänzle, Michael G

    2012-03-01

    This study evaluated the high pressure inactivation of Campylobacter jejuni, Escherichia coli, and poultry meat spoilage organisms. All treatments were performed in aseptically prepared minced poultry meat. Treatment of 19 strains of C. jejuni at 300 MPa and 30°C revealed a large variation of pressure resistance. The recovery of pressure-induced sublethally injured C. jejuni depended on the availability of iron. The addition of iron content to enumeration media was required for resuscitation of sublethally injured cells. Survival of C. jejuni during storage of refrigerated poultry meat was analyzed in fresh and pressuretreated poultry meat, and in the presence or absence of spoilage microbiota. The presence of spoilage microbiota did not significantly influence the survival of C. jejuni. Pressure treatment at 400 MPa and 40°C reduced cell counts of Brochothrix thermosphacta, Carnobacterium divergens, C. jejuni, and Pseudomonas fluorescens to levels below the detection limit. Cell counts of E. coli AW1.7, however, were reduced by only 3.5 log (CFU/g) and remained stable during subsequent refrigerated storage. The resistance to treatment at 600 MPa and 40°C of E. coli AW1.7 was compared with Salmonella enterica, Shiga toxin-producing E. coli and nonpathogenic E. coli strains, and Staphylococcus spp. Cell counts of all organisms except E. coli AW 1.7 were reduced by more than 6 log CFU/g. Cell counts of E. coli AW1.7 were reduced by 4.5 log CFU/g only. Moreover, the ability of E. coli AW1.7 to resist pressure was comparable to the pressure-resistant mutant E. coli LMM1030. Our results indicate that preservation of fresh meat requires a combination of high pressure with high temperature (40 to 60°C) or other antimicrobial hurdles.

  1. Campylobacter jejuni host tissue tropism: a consequence of its low-carb lifestyle?

    PubMed

    Thompson, Stuart A; Gaynor, Erin C

    2008-11-13

    Mechanisms underlying virulence properties of Campylobacter jejuni have historically been difficult to identify. In this issue of Cell Host & Microbe, Hofreuter et al. (2008) show that C. jejuni's ability to metabolize glutamine, glutathione, and asparagine affects its ability to colonize specific host tissues. These findings reflect the emerging theme of bacterial physiology directly impacting pathogenesis.

  2. Antibacterial activity and mechanism of action of zinc oxide nanoparticles against Campylobacter jejuni.

    PubMed

    Xie, Yanping; He, Yiping; Irwin, Peter L; Jin, Tony; Shi, Xianming

    2011-04-01

    The antibacterial effect of zinc oxide (ZnO) nanoparticles on Campylobacter jejuni was investigated for inhibition and inactivation of cell growth. The results showed that C. jejuni was extremely sensitive to treatment with ZnO nanoparticles. The MIC of ZnO nanoparticles for C. jejuni was determined to be 0.05 to 0.025 mg/ml, which is 8- to 16-fold lower than that for Salmonella enterica serovar Enteritidis and Escherichia coli O157:H7 (0.4 mg/ml). The action of ZnO nanoparticles against C. jejuni was determined to be bactericidal, not bacteriostatic. Scanning electron microscopy examination revealed that the majority of the cells transformed from spiral shapes into coccoid forms after exposure to 0.5 mg/ml of ZnO nanoparticles for 16 h, which is consistent with the morphological changes of C. jejuni under other stress conditions. These coccoid cells were found by ethidium monoazide-quantitative PCR (EMA-qPCR) to have a certain level of membrane leakage. To address the molecular basis of ZnO nanoparticle action, a large set of genes involved in cell stress response, motility, pathogenesis, and toxin production were selected for a gene expression study. Reverse transcription-quantitative PCR (RT-qPCR) showed that in response to treatment with ZnO nanoparticles, the expression levels of two oxidative stress genes (katA and ahpC) and a general stress response gene (dnaK) were increased 52-, 7-, and 17-fold, respectively. These results suggest that the antibacterial mechanism of ZnO nanoparticles is most likely due to disruption of the cell membrane and oxidative stress in Campylobacter.

  3. Campylobacter jejuni inactivation in New Zealand soils.

    PubMed

    Ross, C M; Donnison, A M

    2006-11-01

    The study was undertaken to determine the inactivation rate of Campylobacter jejuni in New Zealand soils. Farm dairy effluent (FDE) inoculated at c. 10(5) ml(-1) with C. jejuni was applied to intact soil cores at a rate of 2 l m(-2). Four soils were used: Hamilton (granular); Taupo (pumice); Horotiu and Waihou (allophanic). After FDE application cores were incubated at 10 degrees C for up to 32 days. For all four soils all the FDE remained within the cores and at least 99% of C. jejuni were retained in the top 5 cm. Campylobacter jejuni had declined to the limit of detection (two C. jejuni 100 g(-1)) by 25 days in Hamilton and Taupo soils and by 32 days in Waihou soil. In contrast, in Horotiu soil the decline was only three orders of magnitude after 32 days. Simulated heavy rainfall was applied 4 and 11 days after FDE application and only about 1% of the applied C. jejuni were recovered in leachates. This study demonstrated that at least 99% of applied C. jejuni were retained in the top 5 cm of four soils where they survived for at least 25 days at 10 degrees C. Soil retention of C. jejuni is efficient at FDE application rates that prevent drainage losses. The low infectious dose of C. jejuni and its ability to survive up to 25 days have implications for stock management on dairy farms.

  4. Nutrient Acquisition and Metabolism by Campylobacter jejuni

    PubMed Central

    Stahl, Martin; Butcher, James; Stintzi, Alain

    2012-01-01

    The gastrointestinal pathogen Campylobacter jejuni is able to colonize numerous different hosts and compete against the gut microbiota. To do this, it must be able to efficiently acquire sufficient nutrients from its environment to support its survival and rapid growth in the intestine. However, despite almost 50 years of research, many aspects as to how C. jejuni accomplishes this feat remain poorly understood. C. jejuni lacks many of the common metabolic pathways necessary for the use of glucose, galactose, or other carbohydrates upon which most other microbes thrive. It does however make efficient use of citric acid cycle intermediates and various amino acids. C. jejuni readily uses the amino acids aspartate, glutamate, serine, and proline, with certain strains also possessing additional pathways allowing for the use of glutamine and asparagine. More recent work has revealed that some C. jejuni strains can metabolize the sugar l-fucose. This finding has upset years of dogma that C. jejuni is an asaccharolytic organism. C. jejuni also possesses diverse mechanisms for the acquisition of various transition metals that are required for metabolic activities. In particular, iron acquisition is critical for the formation of iron–sulfur complexes. C. jejuni is also unique in possessing both molybdate and tungsten cofactored proteins and thus has an unusual regulatory scheme for these metals. Together these various metabolic and acquisition pathways help C. jejuni to compete and thrive in wide variety of hosts and environments. PMID:22919597

  5. Complete genomic sequence of campylobacter jejuni subsp. jejuni HS:19 penner reference strain

    USDA-ARS?s Scientific Manuscript database

    Campylobacter jejuni subsp. jejuni (Cjj) infections are a leading cause of foodborne gastroenteritis and the most prevalent antecedent to Guillain-Barré syndrome (GBS). Capsular type Penner HS:19 is among several capsule types shown to be markers for GBS. This study describes the genome of Cjj HS:19...

  6. Campylobacter jejuni biofilms contain extracellular DNA and are sensitive to DNase I treatment

    PubMed Central

    Brown, Helen L.; Hanman, Kate; Reuter, Mark; Betts, Roy P.; van Vliet, Arnoud H. M.

    2015-01-01

    Biofilms make an important contribution to survival and transmission of bacterial pathogens in the food chain. The human pathogen Campylobacter jejuni is known to form biofilms in vitro in food chain-relevant conditions, but the exact roles and composition of the extracellular matrix are still not clear. Extracellular DNA has been found in many bacterial biofilms and can be a major component of the extracellular matrix. Here we show that extracellular DNA is also an important component of the C. jejuni biofilm when attached to stainless steel surfaces, in aerobic conditions and on conditioned surfaces. Degradation of extracellular DNA by exogenous addition of DNase I led to rapid biofilm removal, without loss of C. jejuni viability. Following treatment of a surface with DNase I, C. jejuni was unable to re-establish a biofilm population within 48 h. Similar results were obtained by digesting extracellular DNA with restriction enzymes, suggesting the need for high molecular weight DNA. Addition of C. jejuni genomic DNA containing an antibiotic resistance marker resulted in transfer of the antibiotic resistance marker to susceptible cells in the biofilm, presumably by natural transformation. Taken together, this suggest that eDNA is not only an important component of C. jejuni biofilms and subsequent food chain survival of C. jejuni, but may also contribute to the spread of antimicrobial resistance in C. jejuni. The degradation of extracellular DNA with enzymes such as DNase I is a rapid method to remove C. jejuni biofilms, and is likely to potentiate the activity of antimicrobial treatments and thus synergistically aid disinfection treatments. PMID:26217328

  7. Campylobacter jejuni biofilms contain extracellular DNA and are sensitive to DNase I treatment.

    PubMed

    Brown, Helen L; Hanman, Kate; Reuter, Mark; Betts, Roy P; van Vliet, Arnoud H M

    2015-01-01

    Biofilms make an important contribution to survival and transmission of bacterial pathogens in the food chain. The human pathogen Campylobacter jejuni is known to form biofilms in vitro in food chain-relevant conditions, but the exact roles and composition of the extracellular matrix are still not clear. Extracellular DNA has been found in many bacterial biofilms and can be a major component of the extracellular matrix. Here we show that extracellular DNA is also an important component of the C. jejuni biofilm when attached to stainless steel surfaces, in aerobic conditions and on conditioned surfaces. Degradation of extracellular DNA by exogenous addition of DNase I led to rapid biofilm removal, without loss of C. jejuni viability. Following treatment of a surface with DNase I, C. jejuni was unable to re-establish a biofilm population within 48 h. Similar results were obtained by digesting extracellular DNA with restriction enzymes, suggesting the need for high molecular weight DNA. Addition of C. jejuni genomic DNA containing an antibiotic resistance marker resulted in transfer of the antibiotic resistance marker to susceptible cells in the biofilm, presumably by natural transformation. Taken together, this suggest that eDNA is not only an important component of C. jejuni biofilms and subsequent food chain survival of C. jejuni, but may also contribute to the spread of antimicrobial resistance in C. jejuni. The degradation of extracellular DNA with enzymes such as DNase I is a rapid method to remove C. jejuni biofilms, and is likely to potentiate the activity of antimicrobial treatments and thus synergistically aid disinfection treatments.

  8. Lactobacillus johnsonii ameliorates intestinal, extra-intestinal and systemic pro-inflammatory immune responses following murine Campylobacter jejuni infection.

    PubMed

    Bereswill, Stefan; Ekmekciu, Ira; Escher, Ulrike; Fiebiger, Ulrike; Stingl, Kerstin; Heimesaat, Markus M

    2017-05-18

    Campylobacter jejuni infections are progressively increasing worldwide. Probiotic treatment might open novel therapeutic or even prophylactic approaches to combat campylobacteriosis. In the present study secondary abiotic mice were generated by broad-spectrum antibiotic treatment and perorally reassociated with a commensal murine Lactobacillus johnsonii strain either 14 days before (i.e. prophylactic regimen) or 7 days after (i.e. therapeutic regimen) peroral C. jejuni strain 81-176 infection. Following peroral reassociation both C. jejuni and L. johnsonii were able to stably colonize the murine intestinal tract. Neither therapeutic nor prophylactic L. johnsonii application, however, could decrease intestinal C. jejuni burdens. Notably, C. jejuni induced colonic apoptosis could be ameliorated by prophylactic L. johnsonii treatment, whereas co-administration of L. johnsonii impacted adaptive (i.e. T and B lymphocytes, regulatory T cells), but not innate (i.e. macrophages and monocytes) immune cell responses in the intestinal tract. Strikingly, C. jejuni induced intestinal, extra-intestinal and systemic secretion of pro-inflammatory mediators (such as IL-6, MCP-1, TNF and nitric oxide) could be alleviated by peroral L. johnsonii challenge. In conclusion, immunomodulatory probiotic species might offer valuable strategies for prophylaxis and/or treatment of C. jejuni induced intestinal, extra-intestinal as well as systemic pro-inflammatory immune responses in vivo.

  9. Anti-infective bovine colostrum oligosaccharides: Campylobacter jejuni as a case study.

    PubMed

    Lane, Jonathan A; Mariño, Karina; Naughton, Julie; Kavanaugh, Devon; Clyne, Marguerite; Carrington, Stephen D; Hickey, Rita M

    2012-07-02

    Campylobacter jejuni is the leading cause of acute bacterial infectious diarrhea in humans. Unlike in humans, C. jejuni is a commensal within the avian host. Heavily colonized chickens often fail to display intestinal disease, and no cellular attachment or invasion has been demonstrated in-vivo. Recently, researchers have shown that the reason for the attenuation of C. jejuni virulence may be attributed to the presence of chicken intestinal mucus and more specifically chicken mucin. Since mucins are heavily glycosylated molecules this observation would suggest that glycan-based compounds may act as anti-infectives against C. jejuni. Considering this, we have investigated naturally sourced foods for potential anti-infective glycans. Bovine colostrum rich in neutral and acidic oligosaccharides has been identified as a potential source of anti-infective glycans. In this study, we tested oligosaccharides isolated and purified from the colostrum of Holstein Friesian cows for anti-infective activity against a highly invasive strain of C. jejuni. During our initial studies we structurally defined 37 bovine colostrum oligosaccharides (BCO) by HILIC-HPLC coupled with exoglycosidase digests and off-line mass spectroscopy, and demonstrated the ability of C. jejuni to bind to some of these structures, in-vitro. We also examined the effect of BCO on C. jejuni adhesion to, invasion of and translocation of HT-29 cells. BCO dramatically reduced the cellular invasion and translocation of C. jejuni, in a concentration dependent manner. Periodate treatment of the BCO prior to inhibition studies resulted in a loss of the anti-infective activity of the glycans suggesting a direct oligosaccharide-bacterial interaction. This was confirmed when the BCO completely prevented C. jejuni binding to chicken intestinal mucin, in-vitro. This study builds a strong case for the inclusion of oligosaccharides sourced from cow's milk in functional foods. However, it is only through further

  10. Evaluation of a protective effect of in ovo delivered Campylobacter jejuni OMVs.

    PubMed

    Godlewska, Renata; Kuczkowski, Maciej; Wyszyńska, Agnieszka; Klim, Joanna; Derlatka, Katarzyna; Woźniak-Biel, Anna; Jagusztyn-Krynicka, Elżbieta K

    2016-10-01

    Campylobacter jejuni is the most prevalent cause of a food-borne gastroenteritis in the developed world, with poultry being the main source of infection. Campylobacter jejuni, like other Gram-negative bacteria, constitutively releases outer membrane vesicles (OMVs). OMVs are highly immunogenic, can be taken up by mammalian cells, and are easily modifiable by recombinant engineering. We have tested their usefulness for an oral (in ovo) vaccination of chickens. Four groups of 18-day-old chicken embryos (164 animals) underwent injection of wt C. jejuni OMVs or modified OMVs or PBS into the amniotic fluid. The OMVs modifications relied on overexpression of either a complete wt cjaA gene or the C20A mutant that relocates to the periplasm. Fourteen days post-hatch chicks were orally challenged with live C. jejuni strain. Cecum colonization parameters were analyzed by two-way ANOVA with Tukey post-hoc test. The wtOMVs and OMVs with wtCjaA overexpression were found to confer significant protection of chicken against C. jejuni (p = 0.03 and p = 0.013, respectively) in comparison to PBS controls and are promising candidates for further in ovo vaccine development.

  11. Influence of oral application of Enterococcus faecium AL41 on TGF-β4 and IL-17 expression and immunocompetent cell distribution in chickens challenged with Campylobacter jejuni.

    PubMed

    Letnická, Alica; Karaffová, Viera; Levkut, Mikuláš; Revajová, Viera; Herich, Róbert

    2017-09-01

    Campylobacteriosis is mainly caused by infection with Campylobacter jejuni following consumption or handling of Campylobacter-contaminated poultry meat. The aim of this study was to investigate the effect of probiotic Enterococcus faecium AL41 on TGF-β4 and IL-17 expression and on immunocompetent cell distribution after C. jejuni infection in broiler chicken, as a second part of the previous study of Karaffová et al. (2017). Accordingly, day-old chicks were randomly divided into four experimental groups of 10 chicks each (n = 10): control (C), E. faecium AL41 (EFAL41), C. jejuni CCM6191 (CJ), and combined E. faecium AL41 + C. jejuni CCM6191 (EFAL41 + CJ). Samples from the caecum were collected on days 4 and 7 post Campylobacter infection (dpi), for the isolation of mRNA of TGF-β4, IL-17 and for immunohistochemistry. The relative mRNA expression of TGF-β4 was upregulated in the combined (EFAL41 + CJ) group compared to other groups during both samplings, but the expression of IL-17 was downregulated. Similarly, the highest density of CD3+ was detected in the combined group at 7 dpi, but the number of IgA+ cells was increased in both groups with EFAL41. It was concluded that the EFAL41 probiotic E. faecium strain can modulate the expression of selected cytokines (upregulation of TGF-β4 but downregulation of IL-17 relative expression), and activate IgA-producing cells in the caeca of chicks infected with C. jejuni CCM6191.

  12. Contamination of meat with Campylobacter jejuni in Saitama, Japan.

    PubMed

    Ono, K; Yamamoto, K

    1999-03-15

    To determine the source of food contamination with Campylobacter jejuni, we investigated retail meat, a chicken processing plant and a broiler farm. C. jejuni was found in domestic retailed poultry (45.8%) and imported poultry (3.7%), but not in beef or pork. In the poultry processing plant, there is significant contamination with C. jejuni in chicken carcasses, equipment and workers' hands. This contamination increases during the defeathering and evisceration processes. RAPD analysis shows that contamination with C. jejuni is of intestinal origin. In a broiler farm, C. jejuni was first isolated from a faecal sample of broiler chicken after the 20th day of age. Two weeks later, all birds in this farm became C. jejuni positive. RAPD analysis indicated that C. jejuni spread rapidly from one broiler flock to the other flocks on the farm.

  13. Campylobacter jejuni in commercial eggs.

    PubMed

    Fonseca, Belchiolina Beatriz; Beletti, Marcelo Emílio; de Melo, Roberta Torres; Mendonça, Eliane Pereira; Coelho, Letícia Ríspoli; Nalevaiko, Priscila Christen; Rossi, Daise Aparecida

    2014-01-01

    This study evaluated the ability of Campylobacter jejuni to penetrate through the pores of the shells of commercial eggs and colonize the interior of these eggs, which may become a risk factor for human infection. Furthermore, this study assessed the survival and viability of the bacteria in commercial eggs. The eggs were placed in contact with wood shavings infected with C. jejuni to check the passage of the bacteria. In parallel, the bacteria were inoculated directly into the air chamber to assess the viability in the egg yolk. To determine whether the albumen and egg fertility interferes with the entry and survival of bacteria, we used varying concentrations of albumen and SPF and commercial eggs. C. jejuni was recovered in SPF eggs (fertile) after three hours in contact with contaminated wood shavings but not in infertile commercial eggs. The colonies isolated in the SPF eggs were identified by multiplex PCR and the similarity between strains verified by RAPD-PCR. The bacteria grew in different concentrations of albumen in commercial and SPF eggs. We did not find C. jejuni in commercial eggs inoculated directly into the air chamber, but the bacteria were viable during all periods tested in the wood shavings. This study shows that consumption of commercial eggs infected with C. jejuni does not represent a potential risk to human health.

  14. Virulence characteristics of five new Campylobacter jejuni chicken isolates.

    PubMed

    Stef, Lavinia; Cean, Ada; Vasile, Aida; Julean, Calin; Drinceanu, Dan; Corcionivoschi, Nicolae

    2013-12-13

    Campylobacter enteritis has emerged as one of the most common forms of human diarrheal illness. In this study we have investigated the virulence potential of five new C. jejuni chicken isolates (RO14, RO19, RO24, RO29 and RO37) originated from private households in the rural regions of Banat and Transylvania in Romania. Following isolation and in vitro virulence assay, on HCT-8 cells, our results show that all the C. jejuni chicken isolates overcome the virulence abilities of the highly virulent strain C. jejuni 81-176. Motility, an important virulence factor was significantly improved in all the new chicken isolates. The ability to survive to the antimicrobial activity of the human serum, to resist to the violent attack of bile acids and to survive in the presence of synthetic antibiotics was increased in all the chicken isolates. However, these were statistically significant only for isolates RO29 and RO37. In conclusion our study shows, based on invasiveness and motility, and also on the data provided by the serum and bile resistance experiments that all the new chicken isolates are able to infect human cells, in vitro, and could potentially represent a health hazard for humans.

  15. Campylobacter jejuni--an emerging foodborne pathogen.

    PubMed

    Altekruse, S F; Stern, N J; Fields, P I; Swerdlow, D L

    1999-01-01

    Campylobacter jejuni is the most commonly reported bacterial cause of foodborne infection in the United States. Adding to the human and economic costs are chronic sequelae associated with C. jejuni infection--Guillian-Barré syndrome and reactive arthritis. In addition, an increasing proportion of human infections caused by C. jejuni are resistant to antimicrobial therapy. Mishandling of raw poultry and consumption of undercooked poultry are the major risk factors for human campylobacteriosis. Efforts to prevent human illness are needed throughout each link in the food chain.

  16. Antimicrobial Activities of Isothiocyanates Against Campylobacter jejuni Isolates

    PubMed Central

    Dufour, Virginie; Alazzam, Bachar; Ermel, Gwennola; Thepaut, Marion; Rossero, Albert; Tresse, Odile; Baysse, Christine

    2012-01-01

    Food-borne human infection with Campylobacter jejuni is a medical concern in both industrialized and developing countries. Efficient eradication of C. jejuni reservoirs within live animals and processed foods is limited by the development of antimicrobial resistances and by practical problems related to the use of conventional antibiotics in food processes. We have investigated the bacteriostatic and bactericidal activities of two phytochemicals, allyl-isothiocyanate (AITC), and benzyl isothiocyanate (BITC), against 24 C. jejuni isolates from chicken feces, human infections, and contaminated foods, as well as two reference strains NCTC11168 and 81-176. AITC and BITC displayed a potent antibacterial activity against C. jejuni. BITC showed a higher overall antibacterial effect (MIC of 1.25–5 μg mL−1) compared to AITC (MIC of 50–200 μg mL−1). Both compounds are bactericidal rather than bacteriostatic. The sensitivity levels of C. jejuni isolates against isothiocyanates were neither correlated with the presence of a GGT (γ-Glutamyl Transpeptidase) encoding gene in the genome, with antibiotic resistance nor with the origin of the biological sample. However the ggt mutant of C. jejuni 81-176 displayed a decreased survival rate compared to wild-type when exposed to ITC. This work determined the MIC of two ITC against a panel of C. jejuni isolates, showed that both compounds are bactericidal rather than bacteriostatic, and highlighted the role of GGT enzyme in the survival rate of C. jejuni exposed to ITC. PMID:22919644

  17. Characterization of the biochemical properties of Campylobacter jejuni RNase III

    PubMed Central

    Haddad, Nabila; Saramago, Margarida; Matos, Rute G.; Prévost, Hervé; Arraiano, Cecília M.

    2013-01-01

    Campylobacter jejuni is a foodborne bacterial pathogen, which is now considered as a leading cause of human bacterial gastroenteritis. The information regarding ribonucleases in C. jejuni is very scarce but there are hints that they can be instrumental in virulence mechanisms. Namely, PNPase (polynucleotide phosphorylase) was shown to allow survival of C. jejuni in refrigerated conditions, to facilitate bacterial swimming, cell adhesion, colonization and invasion. In several microorganisms PNPase synthesis is auto-controlled in an RNase III (ribonuclease III)-dependent mechanism. Thereby, we have cloned, overexpressed, purified and characterized Cj-RNase III (C. jejuni RNase III). We have demonstrated that Cj-RNase III is able to complement an Escherichia coli rnc-deficient strain in 30S rRNA processing and PNPase regulation. Cj-RNase III was shown to be active in an unexpectedly large range of conditions, and Mn2+ seems to be its preferred co-factor, contrarily to what was described for other RNase III orthologues. The results lead us to speculate that Cj-RNase III may have an important role under a Mn2+-rich environment. Mutational analysis strengthened the function of some residues in the catalytic mechanism of action of RNase III, which was shown to be conserved. PMID:24073828

  18. L-Fucose metabolism in camplobacter jejuni

    USDA-ARS?s Scientific Manuscript database

    Campylobacter jejuni is a gastrointestinal pathogen once considered asaccharolytic, but now known to metabolize fucose. Strains with the fuc locus encode enzymes for fucose uptake and metabolism and show a competitive colonization advantage in the piglet disease model. C. jejuni NCTC11168 shows redu...

  19. Can microbiota transplantation abrogate murine colonization resistance against Campylobacter jejuni?

    PubMed

    Heimesaat, M M; Plickert, R; Fischer, A; Göbel, U B; Bereswill, S

    2013-03-01

    Enterocolitis caused by Campylobacter jejuni represents an important socioeconomic burden worldwide. The host-specific intestinal microbiota is essential for maintaining colonization resistance (CR) against C. jejuni in conventional mice. Notably, CR is abrogated by shifts of the intestinal microbiota towards overgrowth with commensal E. coli during acute ileitis. Thus, we investigated whether oral transplantation (TX) of ileal microbiota derived from C. jejuni susceptible mice with acute ileitis overcomes CR of healthy conventional animals. Four days following ileitis microbiota TX or ileitis induction and right before C. jejuni infection, mice displayed comparable loads of main intestinal bacterial groups as shown by culture. Eight days following ileitis induction, but not ileal microbiota TX, however, C. jejuni could readily colonize the gastrointestinal tract of conventional mice and also translocate to extra-intestinal tissue sites such as mesenteric lymph nodes, spleen, liver, and blood within 4 days following oral infection. Of note, C. jejuni did not further deteriorate histopathology following ileitis induction. Lack of C. jejuni colonization in TX mice was accompanied by a decrease of commensal E. coli loads in the feces 4 days following C. jejuni infection. In summary, oral ileal microbiota TX from susceptible donors is not sufficient to abrogate murine CR against C. jejuni.

  20. Environmental Stress-Induced Bacterial Lysis and Extracellular DNA Release Contribute to Campylobacter jejuni Biofilm Formation.

    PubMed

    Feng, Jinsong; Ma, Lina; Nie, Jiatong; Konkel, Michael E; Lu, Xiaonan

    2018-03-01

    Campylobacter jejuni is a microaerophilic bacterium and is believed to persist in a biofilm to antagonize environmental stress. This study investigated the influence of environmental conditions on the formation of C. jejuni biofilm. We report an extracellular DNA (eDNA)-mediated mechanism of biofilm formation in response to aerobic and starvation stress. The eDNA was determined to represent a major form of constitutional material of C. jejuni biofilms and to be closely associated with bacterial lysis. Deletion mutation of the stress response genes spoT and recA enhanced the aerobic influence by stimulating lysis and increasing eDNA release. Flagella were also involved in biofilm formation but mainly contributed to attachment rather than induction of lysis. The addition of genomic DNA from either Campylobacter or Salmonella resulted in a concentration-dependent stimulation effect on biofilm formation, but the effect was not due to forming a precoating DNA layer. Enzymatic degradation of DNA by DNase I disrupted C. jejuni biofilm. In a dual-species biofilm, eDNA allocated Campylobacter and Salmonella at distinct spatial locations that protect Campylobacter from oxygen stress. Our findings demonstrated an essential role and multiple functions of eDNA in biofilm formation of C. jejuni , including facilitating initial attachment, establishing and maintaining biofilm, and allocating bacterial cells. IMPORTANCE Campylobacter jejuni is a major cause of foodborne illness worldwide. In the natural environment, the growth of C. jejuni is greatly inhibited by various forms of environmental stress, such as aerobic stress and starvation stress. Biofilm formation can facilitate the distribution of C. jejuni by enabling the survival of this fragile microorganism under unfavorable conditions. However, the mechanism of C. jejuni biofilm formation in response to environmental stress has been investigated only partially. The significance of our research is in identifying extracellular

  1. Extreme Heat Resistance of Food Borne Pathogens Campylobacter jejuni, Escherichia coli, and Salmonella typhimurium on Chicken Breast Fillet during Cooking

    PubMed Central

    de Jong, Aarieke E. I.; van Asselt, Esther D.; Zwietering, Marcel H.; Nauta, Maarten J.; de Jonge, Rob

    2012-01-01

    The aim of this research was to determine the decimal reduction times of bacteria present on chicken fillet in boiling water. The experiments were conducted with Campylobacter jejuni, Salmonella, and Escherichia coli. Whole chicken breast fillets were inoculated with the pathogens, stored overnight (4°C), and subsequently cooked. The surface temperature reached 70°C within 30 sec and 85°C within one minute. Extremely high decimal reduction times of 1.90, 1.97, and 2.20 min were obtained for C. jejuni, E. coli, and S. typhimurium, respectively. Chicken meat and refrigerated storage before cooking enlarged the heat resistance of the food borne pathogens. Additionally, a high challenge temperature or fast heating rate contributed to the level of heat resistance. The data were used to assess the probability of illness (campylobacteriosis) due to consumption of chicken fillet as a function of cooking time. The data revealed that cooking time may be far more critical than previously assumed. PMID:22389647

  2. Accessory genetic content in Campylobacter jejuni ST21CC isolates from feces and blood.

    PubMed

    Skarp, C P A; Akinrinade, O; Kaden, R; Johansson, C; Rautelin, H

    2017-06-01

    Campylobacter jejuni is an important foodborne pathogen and the most commonly reported bacterial cause of gastroenteritis. C. jejuni is occasionally found in blood, although mechanisms important for invasiveness have remained unclear. C. jejuni is divided into many different lineages, of which the ST21 clonal complex (CC) is widely distributed. Here, we performed comparative genomic and in vitro analyses on 17C. jejuni ST21CC strains derived from human blood and feces in order to identify features associated with isolation site. The ST21CC lineage is divided into two large groups; centered around ST-21 and ST-50. Our clinical strains, typed as ST-50, showed further microevolution into two distinct clusters. These clusters were distinguished by major differences in their capsule loci and the distribution of accessory genetic content, including C. jejuni integrated elements (CJIEs) and plasmids. Accessory genetic content was more common among fecal than blood strains, whereas blood strains contained a hybrid capsule locus which partially consisted of C. jejuni subsp. doylei-like content. In vitro infection assays with human colon cell lines did not show significant differences in adherence and invasion between the blood and fecal strains. Our results showed that CJIEs and plasmid derived genetic material were less common among blood isolates than fecal isolates; in contrast, hybrid capsule loci, especially those containing C. jejuni subsp. doylei-like gene content, were found among many isolates derived from blood. The role of these findings requires more detailed investigation. Copyright © 2017 The Authors. Published by Elsevier GmbH.. All rights reserved.

  3. Trans-Cinnamaldehyde, Carvacrol, and Eugenol Reduce Campylobacter jejuni Colonization Factors and Expression of Virulence Genes in Vitro

    PubMed Central

    Upadhyay, Abhinav; Arsi, Komala; Wagle, Basanta R.; Upadhyaya, Indu; Shrestha, Sandip; Donoghue, Ann M.; Donoghue, Dan J.

    2017-01-01

    Campylobacter jejuni is a major foodborne pathogen that causes severe gastroenteritis in humans characterized by fever, diarrhea, and abdominal cramps. In the human gut, Campylobacter adheres and invades the intestinal epithelium followed by cytolethal distending toxin mediated cell death, and enteritis. Reducing the attachment and invasion of Campylobacter to intestinal epithelium and expression of its virulence factors such as motility and cytolethal distending toxin (CDT) production could potentially reduce infection in humans. This study investigated the efficacy of sub-inhibitory concentrations (SICs, concentration not inhibiting bacterial growth) of three GRAS (generally recognized as safe) status phytochemicals namely trans-cinnamaldehyde (TC; 0.005, 0.01%), carvacrol (CR; 0.001, 0.002%), and eugenol (EG; 0.005, 0.01%) in reducing the attachment, invasion, and translocation of C. jejuni on human intestinal epithelial cells (Caco-2). Additionally, the effect of these phytochemicals on Campylobacter motility and CDT production was studied using standard bioassays and gene expression analysis. All experiments had duplicate samples and were replicated three times on three strains (wild type S-8, NCTC 11168, 81–176) of C. jejuni. Data were analyzed using ANOVA with GraphPad ver. 6. Differences between the means were considered significantly different at P < 0.05. The majority of phytochemical treatments reduced C. jejuni adhesion, invasion, and translocation of Caco-2 cells (P < 0.05). In addition, the phytochemicals reduced pathogen motility and production of CDT in S-8 and NCTC 11168 (P < 0.05). Real-time quantitative PCR revealed that phytochemicals reduced the transcription of select C. jejuni genes critical for infection in humans (P < 0.05). Results suggest that TC, CR, and EG could potentially be used to control C. jejuni infection in humans. PMID:28487683

  4. Whole Genome Sequencing and Multiplex qPCR Methods to Identify Campylobacter jejuni Encoding cst-II or cst-III Sialyltransferase

    PubMed Central

    Neal-McKinney, Jason M.; Liu, Kun C.; Jinneman, Karen C.; Wu, Wen-Hsin; Rice, Daniel H.

    2018-01-01

    Campylobacter jejuni causes more than 2 million cases of gastroenteritis annually in the United States, and is also linked to the autoimmune sequelae Guillan–Barre syndrome (GBS). GBS often results in flaccid paralysis, as the myelin sheaths of nerve cells are degraded by the adaptive immune response. Certain strains of C. jejuni modify their lipooligosaccharide (LOS) with the addition of neuraminic acid, resulting in LOS moieties that are structurally similar to gangliosides present on nerve cells. This can trigger GBS in a susceptible host, as antibodies generated against C. jejuni can cross-react with gangliosides, leading to demyelination of nerves and a loss of signal transduction. The goal of this study was to develop a quantitative PCR (qPCR) method and use whole genome sequencing data to detect the Campylobacter sialyltransferase (cst) genes responsible for the addition of neuraminic acid to LOS. The qPCR method was used to screen a library of 89 C. jejuni field samples collected by the Food and Drug Administration Pacific Northwest Lab (PNL) as well as clinical isolates transferred to PNL. In silico analysis was used to screen 827 C. jejuni genomes in the FDA GenomeTrakr SRA database. The results indicate that a majority of C. jejuni strains could produce LOS with ganglioside mimicry, as 43.8% of PNL isolates and 46.9% of the GenomeTrakr isolates lacked the cst genes. The methods described in this study can be used by public health laboratories to rapidly determine whether a C. jejuni isolate has the potential to induce GBS. Based on these results, a majority of C. jejuni in the PNL collection and submitted to GenomeTrakr have the potential to produce LOS that mimics human gangliosides. PMID:29615986

  5. Whole Genome Sequencing and Multiplex qPCR Methods to Identify Campylobacter jejuni Encoding cst-II or cst-III Sialyltransferase.

    PubMed

    Neal-McKinney, Jason M; Liu, Kun C; Jinneman, Karen C; Wu, Wen-Hsin; Rice, Daniel H

    2018-01-01

    Campylobacter jejuni causes more than 2 million cases of gastroenteritis annually in the United States, and is also linked to the autoimmune sequelae Guillan-Barre syndrome (GBS). GBS often results in flaccid paralysis, as the myelin sheaths of nerve cells are degraded by the adaptive immune response. Certain strains of C. jejuni modify their lipooligosaccharide (LOS) with the addition of neuraminic acid, resulting in LOS moieties that are structurally similar to gangliosides present on nerve cells. This can trigger GBS in a susceptible host, as antibodies generated against C. jejuni can cross-react with gangliosides, leading to demyelination of nerves and a loss of signal transduction. The goal of this study was to develop a quantitative PCR (qPCR) method and use whole genome sequencing data to detect the Campylobacter sialyltransferase ( cst ) genes responsible for the addition of neuraminic acid to LOS. The qPCR method was used to screen a library of 89 C. jejuni field samples collected by the Food and Drug Administration Pacific Northwest Lab (PNL) as well as clinical isolates transferred to PNL. In silico analysis was used to screen 827 C. jejuni genomes in the FDA GenomeTrakr SRA database. The results indicate that a majority of C. jejuni strains could produce LOS with ganglioside mimicry, as 43.8% of PNL isolates and 46.9% of the GenomeTrakr isolates lacked the cst genes. The methods described in this study can be used by public health laboratories to rapidly determine whether a C. jejuni isolate has the potential to induce GBS. Based on these results, a majority of C. jejuni in the PNL collection and submitted to GenomeTrakr have the potential to produce LOS that mimics human gangliosides.

  6. Pseudomonas aeruginosa facilitates Campylobacter jejuni growth in biofilms under oxic flow conditions.

    PubMed

    Culotti, Alessandro; Packman, Aaron I

    2015-12-01

    We investigated the growth of Campylobacter jejuni in biofilms with Pseudomonas aeruginosa under oxic flow conditions. We observed the growth of C. jejuni in mono-culture, deposited on pre-established P. aeruginosa biofilms, and co-inoculated with P. aeruginosa. In mono-culture, C. jejuni was unable to form biofilms. However, deposited C. jejuni continuously grew on pre-established P. aeruginosa biofilms for a period of 3 days. The growth of scattered C. jejuni clusters was strictly limited to the P. aeruginosa biofilm surface, and no intergrowth was observed. Co-culturing of C. jejuni and P. aeruginosa also enabled the growth of both organisms in biofilms, with C. jejuni clusters developing on the surface of the P. aeruginosa biofilm. Dissolved oxygen (DO) measurements in the medium showed that P. aeruginosa biofilms depleted the effluent DO from 9.0 to 0.5 mg L(-1) 24 hours after inoculation. The localized microaerophilic environment generated by P. aeruginosa promoted the persistence and growth of C. jejuni. Our findings show that P. aeruginosa not only prolongs the survival of C. jejuni under oxic conditions, but also enables the growth of C. jejuni on the surface of P. aeruginosa biofilms. © FEMS 2015. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  7. Adherence Reduction of Campylobacter jejuni and Campylobacter coli Strains to HEp-2 Cells by Mannan Oligosaccharides and a High-Molecular-Weight Component of Cranberry Extract.

    PubMed

    Ramirez-Hernandez, Alejandra; Rupnow, John; Hutkins, Robert W

    2015-08-01

    Campylobacter infections are a leading cause of human bacterial gastroenteritis in the United States and are a major cause of diarrheal disease throughout the world. Colonization and subsequent infection and invasion of Campylobacter require that the bacteria adhere to the surface of host cells. Agents that inhibit adherence could be used prophylactically to reduce Campylobacter carriage and infection. Mannan oligosaccharides (MOS) have been used as a feed supplement in livestock animals to improve performance and to replace growth-promoting antibiotics. However, MOS and other nondigestible oligosaccharides may also prevent pathogen colonization by inhibiting adherence in the gastrointestinal tract. In addition, plant extracts, including those derived from cranberries, have been shown to have antiadherence activity against pathogens. The goal of this study was to assess the ability of MOS and cranberry fractions to serve as antiadherence agents against strains of Campylobacter jejuni and Campylobacter coli. Adherence experiments were performed using HEp-2 cells. Significant reductions in adherence of C. jejuni 29438, C. jejuni 700819, C. jejuni 3329, and C. coli 43485 were observed in the presence of MOS (up to 40 mg/ml) and with a high-molecular-weight fraction of cranberry extract (up to 3 mg/ml). However, none of the tested materials reduced adherence of C. coli BAA-1061. No additive effect in adherence inhibition was observed for an MOS-cranberry blend. These results suggest that both components, MOS and cranberry, could be used to reduce Campylobacter colonization and carriage in livestock animals and potentially limit human exposure to this pathogen.

  8. Intrinsic and Extrinsic Aspects on Campylobacter jejuni Biofilms

    PubMed Central

    Melo, Roberta T.; Mendonça, Eliane P.; Monteiro, Guilherme P.; Siqueira, Mariana C.; Pereira, Clara B.; Peres, Phelipe A. B. M.; Fernandez, Heriberto; Rossi, Daise A.

    2017-01-01

    Biofilm represents a way of life that allows greater survival of microorganisms in hostile habitats. Campylobacter jejuni is able to form biofilms in vitro and on surfaces at several points in the poultry production chain. Genetic determinants related to their formation are expressed differently between strains and external conditions are decisive in this respect. Our approach combines phylogenetic analysis and the presence of seven specific genes linked to biofilm formation in association with traditional microbiology techniques, using Mueller Hinton and chicken juice as substrates in order to quantify, classify, determine the composition and morphology of the biomass of simple and mixed biofilms of 30 C. jejuni strains. It also evaluates the inhibition of its formation by biocides commonly used in industry and also by zinc oxide nanoparticles. Genetic analysis showed high heterogeneity with the identification of 23 pulsotypes. Despite the diversity, the presence of flaA, cadF, luxS, dnaJ, htrA, cbrA, and sodB genes in all strains shows the high potential for biofilm formation. This ability was only expressed in chicken juice, where they presented phenotype of a strong biofilm producer, with a mean count of 7.37 log CFU/mL and an ultrastructure characteristic of mature biofilm. The composition of simple and mixed biofilms was predominantly composed by proteins. The exceptions were found in mixed biofilms with Pseudomonas aeruginosa, which includes a carbohydrate-rich matrix, lower ability to sessile form in chicken juice and compact architecture of the biofilm, this aspects are intrinsic to this species. Hypochlorite, chlorhexidine, and peracetic acid were more effective in controlling viable cells of C. jejuni in biofilm, but the existence of tolerant strains indicates exposure to sublethal concentrations and development of adaptation mechanisms. This study shows that in chicken juice C. jejuni presents greater potential in producing mature biofilms. PMID

  9. Intrinsic and Extrinsic Aspects on Campylobacter jejuni Biofilms.

    PubMed

    Melo, Roberta T; Mendonça, Eliane P; Monteiro, Guilherme P; Siqueira, Mariana C; Pereira, Clara B; Peres, Phelipe A B M; Fernandez, Heriberto; Rossi, Daise A

    2017-01-01

    Biofilm represents a way of life that allows greater survival of microorganisms in hostile habitats. Campylobacter jejuni is able to form biofilms in vitro and on surfaces at several points in the poultry production chain. Genetic determinants related to their formation are expressed differently between strains and external conditions are decisive in this respect. Our approach combines phylogenetic analysis and the presence of seven specific genes linked to biofilm formation in association with traditional microbiology techniques, using Mueller Hinton and chicken juice as substrates in order to quantify, classify, determine the composition and morphology of the biomass of simple and mixed biofilms of 30 C. jejuni strains. It also evaluates the inhibition of its formation by biocides commonly used in industry and also by zinc oxide nanoparticles. Genetic analysis showed high heterogeneity with the identification of 23 pulsotypes. Despite the diversity, the presence of flaA, cadF, luxS, dnaJ, htrA, cbrA , and sodB genes in all strains shows the high potential for biofilm formation. This ability was only expressed in chicken juice, where they presented phenotype of a strong biofilm producer, with a mean count of 7.37 log CFU/mL and an ultrastructure characteristic of mature biofilm. The composition of simple and mixed biofilms was predominantly composed by proteins. The exceptions were found in mixed biofilms with Pseudomonas aeruginosa , which includes a carbohydrate-rich matrix, lower ability to sessile form in chicken juice and compact architecture of the biofilm, this aspects are intrinsic to this species. Hypochlorite, chlorhexidine, and peracetic acid were more effective in controlling viable cells of C. jejuni in biofilm, but the existence of tolerant strains indicates exposure to sublethal concentrations and development of adaptation mechanisms. This study shows that in chicken juice C. jejuni presents greater potential in producing mature biofilms.

  10. Reduction of Campylobacter jejuni on chicken wings by chemical treatments.

    PubMed

    Zhao, Tong; Doyle, Michael P

    2006-04-01

    Eight chemicals, including glycerol monolaurate, hydrogen peroxide, acetic acid, lactic acid, sodium benzoate, sodium chlorate, sodium carbonate, and sodium hydroxide, were tested individually or in combination for their ability to inactivate Campylobacter jejuni at 4 degrees C in suspension. Results showed that treatment for up to 20 min with 0.01% glycerol monolaurate, 0.1% sodium benzoate, 50 or 100 mM sodium chlorate, or 1% lactic acid did not substantially (< or = 0.5 log CFU/ml) reduce C. jejuni populations but that 0.1 and 0.2% hydrogen peroxide for 20 min reduced C. jejuni populations by ca. 2.0 and 4.5 log CFU/ml, respectively. By contrast, treatments with 0.5, 1.0, 1.5, and 2.0% acetic acid, 25, 50, and 100 mM sodium carbonate, and 0.05 and 0.1 N sodium hydroxide reduced C. jejuni populations by >5 log CFU/ml within 2 min. A combination of 0.5% acetic acid plus 0.05% potassium sorbate or 0.5% acetic acid plus 0.05% sodium benzoate reduced C. jejuni populations by >5 log CFU/ml within 1 min; however, substituting 0.5% lactic acid for 0.5% acetic acid was not effective, with a reduction of C. jejuni of <0.5 log CFU/ml. A combination of acidic calcium sulfate, lactic acid, ethanol, sodium dodecyl sulfate, and polypropylene glycol (ACS-LA) also reduced C. jejuni in suspension by >5 log CFU/ml within 1 min. All chemicals or chemical combinations for which there was a >5-log/ml reduction of C. jejuni in suspension were further evaluated for C. jejuni inactivation on chicken wings. Treatments at 4 degrees C of 2% acetic acid, 100 mM sodium carbonate, or 0.1 N sodium hydroxide for up to 45 s reduced C. jejuni populations by ca. 1.4, 1.6, or 3.5 log CFU/g, respectively. Treatment with ACS-LA at 4 degrees C for 15 s reduced C. jejuni by >5 log CFU/g to an undetectable level. The ACS-LA treatment was highly effective in chilled water at killing C. jejuni on chicken and, if recycled, may be a useful treatment in chill water tanks for poultry processors to reduce

  11. A novel O-linked glycan modulates Campylobacter jejuni major outer membrane protein-mediated adhesion to human histo-blood group antigens and chicken colonization

    PubMed Central

    Mahdavi, Jafar; Pirinccioglu, Necmettin; Oldfield, Neil J.; Carlsohn, Elisabet; Stoof, Jeroen; Aslam, Akhmed; Self, Tim; Cawthraw, Shaun A.; Petrovska, Liljana; Colborne, Natalie; Sihlbom, Carina; Borén, Thomas; Wooldridge, Karl G.; Ala'Aldeen, Dlawer A. A.

    2014-01-01

    Campylobacter jejuni is an important cause of human foodborne gastroenteritis; strategies to prevent infection are hampered by a poor understanding of the complex interactions between host and pathogen. Previous work showed that C. jejuni could bind human histo-blood group antigens (BgAgs) in vitro and that BgAgs could inhibit the binding of C. jejuni to human intestinal mucosa ex vivo. Here, the major flagella subunit protein (FlaA) and the major outer membrane protein (MOMP) were identified as BgAg-binding adhesins in C. jejuni NCTC11168. Significantly, the MOMP was shown to be O-glycosylated at Thr268; previously only flagellin proteins were known to be O-glycosylated in C. jejuni. Substitution of MOMP Thr268 led to significantly reduced binding to BgAgs. The O-glycan moiety was characterized as Gal(β1–3)-GalNAc(β1–4)-GalNAc(β1–4)-GalNAcα1-Thr268; modelling suggested that O-glycosylation has a notable effect on the conformation of MOMP and this modulates BgAg-binding capacity. Glycosylation of MOMP at Thr268 promoted cell-to-cell binding, biofilm formation and adhesion to Caco-2 cells, and was required for the optimal colonization of chickens by C. jejuni, confirming the significance of this O-glycosylation in pathogenesis. PMID:24451549

  12. Complete genomic sequence of Campylobacter jejuni subsp. jejuni HS:19 strain RM1285 that was isolated from packaged chicken

    USDA-ARS?s Scientific Manuscript database

    Poultry products serve as the main source of Campylobacter jejuni subsp. jejuni (Cjj) infections in humans. Cjj infections are a leading cause of foodborne gastroenteritis and are a prevalent antecedent to Guillain-Barré syndrome (GBS). This study describes the genome of Cjj HS:19 strain RM1285 isol...

  13. Effects of antibiotic resistance (AR) and microbiota shifts on Campylobacter jejuni-mediated diseases.

    PubMed

    Brooks, Phillip T; Mansfield, Linda S

    2017-12-01

    Campylobacter jejuni is an important zoonotic pathogen recently designated a serious antimicrobial resistant (AR) threat. While most patients with C. jejuni experience hemorrhagic colitis, serious autoimmune conditions can follow including inflammatory bowel disease (IBD) and the acute neuropathy Guillain Barré Syndrome (GBS). This review examines inter-relationships among factors mediating C. jejuni diarrheal versus autoimmune disease especially AR C. jejuni and microbiome shifts. Because both susceptible and AR C. jejuni are acquired from animals or their products, we consider their role in harboring strains. Inter-relationships among factors mediating C. jejuni colonization, diarrheal and autoimmune disease include C. jejuni virulence factors and AR, the enteric microbiome, and host responses. Because AR C. jejuni have been suggested to affect the severity of disease, length of infections and propensity to develop GBS, it is important to understand how these interactions occur when strains are under selection by antimicrobials. More work is needed to elucidate host-pathogen interactions of AR C. jejuni compared with susceptible strains and how AR C. jejuni are maintained and evolve in animal reservoirs and the extent of transmission to humans. These knowledge gaps impair the development of effective strategies to prevent the emergence of AR C. jejuni in reservoir species and human populations.

  14. Survival of cold-stressed Campylobacter jejuni on ground chicken and chicken skin during frozen storage.

    PubMed

    Bhaduri, Saumya; Cottrell, Bryan

    2004-12-01

    Campylobacter jejuni is prevalent in poultry, but the effect of combined refrigerated and frozen storage on its survival, conditions relevant to poultry processing and storage, has not been evaluated. Therefore, the effects of refrigeration at 4 degrees C, freezing at -20 degrees C, and a combination of refrigeration and freezing on the survival of C. jejuni in ground chicken and on chicken skin were examined. Samples were enumerated using tryptic soy agar containing sheep's blood and modified cefoperazone charcoal deoxycholate agar. Refrigerated storage alone for 3 to 7 days produced a reduction in cell counts of 0.34 to 0.81 log10 CFU/g in ground chicken and a reduction in cell counts of 0.31 to 0.63 log10 CFU/g on chicken skin. Declines were comparable for each sample type using either plating medium. Frozen storage, alone and with prerefrigeration, produced a reduction in cell counts of 0.56 to 1.57 log10 CFU/g in ground chicken and a reduction in cell counts of 1.38 to 3.39 log10 CFU/g on chicken skin over a 2-week period. The recovery of C. jejuni following freezing was similar on both plating media. The survival following frozen storage was greater in ground chicken than on chicken skin with or without prerefrigeration. Cell counts after freezing were lower on chicken skin samples that had been prerefrigerated for 7 days than in those that had been prerefrigerated for 0, 1, or 3 days. This was not observed for ground chicken samples, possibly due to their composition. C. jejuni survived storage at 4 and -20 degrees C with either sample type. This study indicates that, individually or in combination, refrigeration and freezing are not a substitute for safe handling and proper cooking of poultry.

  15. Potential virulence and antimicrobial susceptibility of Campylobacter jejuni isolates from food and companion animals.

    PubMed

    Lee, Michelle K; Billington, Stephen J; Joens, Lynn A

    2004-01-01

    Infection in humans with Campylobacter jejuni is commonly associated with exposure to food animal fecal material. In this study, we report on the recovery, potential for virulence and antimicrobial resistance levels of C. jejuni isolated from food and companion animals. Three hundred and seventy-eight fecal samples from food and companion animals and surface swabs from beef carcasses were tested for the presence of C. jejuni. C. jejuni was isolated from 13.8% (11/80) of dogs, 5% (1/20) of goats, 28.3% (17/60) of dairy cattle, 0% (0/65) of range cattle, 73.5% (36/49) of feedlot cattle, and 94.7% (18/19) of beef carcasses. Beef cattle from a single Arizona herd showed a considerable increase in fecal shedding of C. jejuni from pasture to feedlot and over time on the feedlot. Forty-two isolates were tested for susceptibility to four antimicrobial agents, each representing a class of antimicrobial drug approved for use in both humans and animals. None of the isolates were found to be resistant to erythromycin or gentamicin, whereas 2.4% of isolates were resistant to ciprofloxacin and 28.6% of isolates were resistant to tetracycline. The presence of virulence traits among the 42 isolates was assessed using in vitro macrophage survival and epithelial cell adherence and invasion assays. Of the isolates examined, 17 were able to survive within macrophages through 72 h at viable counts of >/=10(3)/well and 12 were capable of invading epithelial cells at viable counts of >/=10(3)/well. Data from these studies suggests that many of the isolates recovered from the non-poultry animal sources have the capacity to cause disease if transmitted to humans.

  16. Campylobacter jejuni transducer like proteins: Chemotaxis and beyond.

    PubMed

    Chandrashekhar, Kshipra; Kassem, Issmat I; Rajashekara, Gireesh

    2017-07-04

    Chemotaxis, a process that mediates directional motility toward or away from chemical stimuli (chemoeffectors/ligands that can be attractants or repellents) in the environment, plays an important role in the adaptation of Campylobacter jejuni to disparate niches. The chemotaxis system consists of core signal transduction proteins and methyl-accepting-domain-containing Transducer like proteins (Tlps). Ligands binding to Tlps relay a signal to chemotaxis proteins in the cytoplasm which initiate a signal transduction cascade, culminating into a directional flagellar movement. Tlps facilitate substrate-specific chemotaxis in C. jejuni, which plays an important role in the pathogen's adaptation, pathobiology and colonization of the chicken gastrointestinal tract. However, the role of Tlps in C. jejuni's host tissue specific colonization, physiology and virulence remains not completely understood. Based on recent studies, it can be predicted that Tlps might be important targets for developing strategies to control C. jejuni via vaccines and antimicrobials.

  17. Phytochemicals reduce biofilm formation and inactivates mature biofilm of Campylobacter jejuni

    USDA-ARS?s Scientific Manuscript database

    Campylobacter jejuni is the leading cause of human foodborne illness globally, and is strongly linked with the consumption of contaminated poultry products. However, little is known about the persistence of C. jejuni in the poultry processing environment. Several studies have shown that C. jejuni ca...

  18. Avian Resistance to Campylobacter jejuni Colonization Is Associated with an Intestinal Immunogene Expression Signature Identified by mRNA Sequencing

    PubMed Central

    Connell, Sarah; Meade, Kieran G.; Allan, Brenda; Lloyd, Andrew T.; Kenny, Elaine; Cormican, Paul; Morris, Derek W.; Bradley, Daniel G.; O'Farrelly, Cliona

    2012-01-01

    Campylobacter jejuni is the most common cause of human bacterial gastroenteritis and is associated with several post-infectious manifestations, including onset of the autoimmune neuropathy Guillain-Barré syndrome, causing significant morbidity and mortality. Poorly-cooked chicken meat is the most frequent source of infection as C. jejuni colonizes the avian intestine in a commensal relationship. However, not all chickens are equally colonized and resistance seems to be genetically determined. We hypothesize that differences in immune response may contribute to variation in colonization levels between susceptible and resistant birds. Using high-throughput sequencing in an avian infection model, we investigate gene expression associated with resistance or susceptibility to colonization of the gastrointestinal tract with C. jejuni and find that gut related immune mechanisms are critical for regulating colonization. Amongst a single population of 300 4-week old chickens, there was clear segregation in levels of C. jejuni colonization 48 hours post-exposure. RNAseq analysis of caecal tissue from 14 C. jejuni-susceptible and 14 C. jejuni-resistant birds generated over 363 million short mRNA sequences which were investigated to identify 219 differentially expressed genes. Significantly higher expression of genes involved in the innate immune response, cytokine signaling, B cell and T cell activation and immunoglobulin production, as well as the renin-angiotensin system was observed in resistant birds, suggesting an early active immune response to C. jejuni. Lower expression of these genes in colonized birds suggests suppression or inhibition of a clearing immune response thus facilitating commensal colonization and generating vectors for zoonotic transmission. This study describes biological processes regulating C. jejuni colonization of the avian intestine and gives insight into the differential immune mechanisms incited in response to commensal bacteria in general

  19. Morphology heterogeneity within a Campylobacter jejuni helical population

    USDA-ARS?s Scientific Manuscript database

    Campylobacter jejuni helical shape is an important factor in colonization and host interactions with straight mutants having altered biological properties. Passage on calcofluor white (CFW) led to the selection of C. jejuni 81-176 isolates with changes in morphology resulting in either a straight mo...

  20. PCR detection of four virulence-associated genes of Campylobacter jejuni isolates from Thai broilers and their abilities of adhesion to and invasion of INT-407 cells.

    PubMed

    Chansiripornchai, Niwat; Sasipreeyajan, Jiroj

    2009-06-01

    Campylobacter jejuni is a major cause of food borne pathogens in humans and a major reservoir for this pathogen is poultry. The C. jejuni in broilers was investigated from in the caeca of broilers. Twenty broiler/flock samples from 7 flocks were assessed. The average prevalence of C. jejuni was 65% in the broiler flocks. The adhesion and invasion ability of 48 strains of C. jejuni on INT 407 were studied. The adhesion and invasion ability of 48 Campylobacter isolates from caecal contents were analyzed with Human embryonic intestine (INT-407) cells being used as a gentamicin resistance assay. The caecal isolates exhibited a wide range of adherence and invasion ability. There was a significant correlation (p<0.01) between the adherence and the invasion ability of the Campylobacter isolates. Each of the virulence-associated genes: dnaJ, cadF, pldA and ciaB was detected by polymerase chain reaction from 100, 76, 31 and 41% of the Campylobacter strains, respectively. All of four virulence-associated genes were detected in 11 isolates. However, there was unclear association between the invasion ability and the presence of virulence-associated genes in this experiment, suggesting that more genes may be involved in the invasion process.

  1. Characterization of the Thermal Stress Response of Campylobacter jejuni

    PubMed Central

    Konkel, Michael E.; Kim, Bong J.; Klena, John D.; Young, Colin R.; Ziprin, Richard

    1998-01-01

    Campylobacter jejuni, a microaerophilic, gram-negative bacterium, is a common cause of gastrointestinal disease in humans. Heat shock proteins are a group of highly conserved, coregulated proteins that play important roles in enabling organisms to cope with physiological stresses. The primary aim of this study was to characterize the heat shock response of C. jejuni. Twenty-four proteins were preferentially synthesized by C. jejuni immediately following heat shock. Upon immunoscreening of Escherichia coli transformants harboring a Campylobacter genomic DNA library, one recombinant plasmid that encoded a heat shock protein was isolated. The recombinant plasmid, designated pMEK20, contained an open reading frame of 1,119 bp that was capable of encoding a protein of 372 amino acids with a calculated molecular mass of 41,436 Da. The deduced amino acid sequence of the open reading frame shared similarity with that of DnaJ, which belongs to the Hsp-40 family of molecular chaperones, from a number of bacteria. An E. coli dnaJ mutant was successfully complemented with the pMEK20 recombinant plasmid, as judged by the ability of bacteriophage λ to form plaques, indicating that the C. jejuni gene encoding the 41-kDa protein is a functional homolog of the dnaJ gene from E. coli. The ability of each of two C. jejuni dnaJ mutants to form colonies at 46°C was severely retarded, indicating that DnaJ plays an important role in C. jejuni thermotolerance. Experiments revealed that a C. jejuni DnaJ mutant was unable to colonize newly hatched Leghorn chickens, suggesting that heat shock proteins play a role in vivo. PMID:9673247

  2. Campylobacter jejuni transducer like proteins: Chemotaxis and beyond

    PubMed Central

    Chandrashekhar, Kshipra; Kassem, Issmat I.; Rajashekara, Gireesh

    2017-01-01

    ABSTRACT Chemotaxis, a process that mediates directional motility toward or away from chemical stimuli (chemoeffectors/ligands that can be attractants or repellents) in the environment, plays an important role in the adaptation of Campylobacter jejuni to disparate niches. The chemotaxis system consists of core signal transduction proteins and methyl-accepting-domain-containing Transducer like proteins (Tlps). Ligands binding to Tlps relay a signal to chemotaxis proteins in the cytoplasm which initiate a signal transduction cascade, culminating into a directional flagellar movement. Tlps facilitate substrate-specific chemotaxis in C. jejuni, which plays an important role in the pathogen's adaptation, pathobiology and colonization of the chicken gastrointestinal tract. However, the role of Tlps in C. jejuni's host tissue specific colonization, physiology and virulence remains not completely understood. Based on recent studies, it can be predicted that Tlps might be important targets for developing strategies to control C. jejuni via vaccines and antimicrobials. PMID:28080213

  3. Gene Expression Profiling of the Local Cecal Response of Genetic Chicken Lines That Differ in Their Susceptibility to Campylobacter jejuni Colonization

    PubMed Central

    Kogut, Michael H.; Chiang, Hsin-I; Wang, Ying; Genovese, Kenneth J.; He, Haiqi; Zhou, Huaijun

    2010-01-01

    Campylobacter jejuni (C. jejuni) is one of the most common causes of human bacterial enteritis worldwide primarily due to contaminated poultry products. Previously, we found a significant difference in C. jejuni colonization in the ceca between two genetically distinct broiler lines (Line A (resistant) has less colony than line B (susceptible) on day 7 post inoculation). We hypothesize that different mechanisms between these two genetic lines may affect their ability to resist C. jejuni colonization in chickens. The molecular mechanisms of the local host response to C. jejuni colonization in chickens have not been well understood. In the present study, to profile the cecal gene expression in the response to C. jejuni colonization and to compare differences between two lines at the molecular level, RNA of ceca from two genetic lines of chickens (A and B) were applied to a chicken whole genome microarray for a pair-comparison between inoculated (I) and non-inoculated (N) chickens within each line and between lines. Our results demonstrated that metabolism process and insulin receptor signaling pathways are key contributors to the different response to C. jejuni colonization between lines A and B. With C. jejuni inoculation, lymphocyte activation and lymphoid organ development functions are important for line A host defenses, while cell differentiation, communication and signaling pathways are important for line B. Interestingly, circadian rhythm appears play a critical role in host response of the more resistant A line to C. jejuni colonization. A dramatic differential host response was observed between these two lines of chickens. The more susceptible line B chickens responded to C. jejuni inoculation with a dramatic up-regulation in lipid, glucose, and amino acid metabolism, which is undoubtedly for use in the response to the colonization with little or no change in immune host defenses. However, in more resistant line A birds the host defense responses were

  4. Impact of oxidative stress defense on bacterial survival and morphological change in Campylobacter jejuni under aerobic conditions.

    PubMed

    Oh, Euna; McMullen, Lynn; Jeon, Byeonghwa

    2015-01-01

    Campylobacter jejuni, a microaerophilic foodborne pathogen, inescapably faces high oxygen tension during its transmission to humans. Thus, the ability of C. jejuni to survive under oxygen-rich conditions may significantly impact C. jejuni viability in food and food safety as well. In this study, we investigated the impact of oxidative stress resistance on the survival of C. jejuni under aerobic conditions by examining three mutants defective in key antioxidant genes, including ahpC, katA, and sodB. All the three mutants exhibited growth reduction under aerobic conditions compared to the wild-type (WT), and the ahpC mutant showed the most significant growth defect. The CFU reduction in the mutants was recovered to the WT level by complementation. Higher levels of reactive oxygen species were accumulated in C. jejuni under aerobic conditions than microaerobic conditions, and supplementation of culture media with an antioxidant recovered the growth of C. jejuni. The levels of lipid peroxidation and protein oxidation were significantly increased in the mutants compared to WT. Additionally, the mutants exhibited different morphological changes under aerobic conditions. The ahpC and katA mutants developed coccoid morphology by aeration, whereas the sodB mutant established elongated cellular morphology. Compared to microaerobic conditions, interestingly, aerobic culture conditions substantially induced the formation of coccoidal cells, and antioxidant treatment reduced the emergence of coccoid forms under aerobic conditions. The ATP concentrations and PMA-qPCR analysis supported that oxidative stress is a factor that induces the development of a viable-but-non-culturable state in C. jejuni. The findings in this study clearly demonstrated that oxidative stress resistance plays an important role in the survival and morphological changes of C. jejuni under aerobic conditions.

  5. Major role for FeoB in Campylobacter jejuni ferrous iron acquisition, gut colonization, and intracellular survival.

    PubMed

    Naikare, Hemant; Palyada, Kiran; Panciera, Roger; Marlow, Denver; Stintzi, Alain

    2006-10-01

    To assess the importance of ferrous iron acquisition in Campylobacter physiology and pathogenesis, we disrupted and characterized the Fe2+ iron transporter, FeoB, in Campylobacter jejuni NCTC 11168, 81-176, and ATCC 43431. The feoB mutant was significantly affected in its ability to transport 55Fe2+. It accumulated half the amount of iron than the wild-type strain during growth in an iron-containing medium. The intracellular iron of the feoB mutant was localized in the periplasmic space versus the cytoplasm for the wild-type strain. These results indicate that the feoB gene of C. jejuni encodes a functional ferrous iron transport system. Reverse transcriptase PCR analysis revealed the cotranscription of feoB and Cj1397, which encodes a homolog of Escherichia coli feoA. C. jejuni 81-176 feoB mutants exhibited reduced ability to persist in human INT-407 embryonic intestinal cells and porcine IPEC-1 small intestinal epithelial cells compared to the wild type. C. jejuni NCTC 11168 feoB mutant was outcompeted by the wild type for colonization and/or survival in the rabbit ileal loop. The feoB mutants of the three C. jejuni strains were significantly affected in their ability to colonize the chick cecum. And finally, the three feoB mutants were outcompeted by their respective wild-type strains for infection of the intestinal tracts of colostrum-deprived piglets. Taken together, these results demonstrate that FeoB-mediated ferrous iron acquisition contributes significantly to colonization of the gastrointestinal tract during both commensal and infectious relationship, and thus it plays an important role in Campylobacter pathogenesis.

  6. [Campylobacter jejuni infections in slaughterhouse workers].

    PubMed

    Mancinelli, S; Riccardi, F; Santi, A L; Palombi, L; Marazzi, M C

    1988-01-01

    Complement fixing (C.F.) antibodies to Campylobacter jejuni were detected in 83 slaughterhouse workers and 83 blood donors. Workers were aged 18-65 years (mean, 41.7 +/- 12.3) and had worked in the slaughterhouse for 2-40 years (mean, 17.5 +/- 5.1). C.F. antibodies were detected according to Mosimann's method and by including five antigens: Campylobacter jejuni, Yersinia enterocolitica types 03 and 09, Yersinia pseudotuberculosis and Brucella. Positive titers were found in 12.1% of workers and in 2.4% of control subjects (p less than 0.01); values ranged from 1:10 to 1:40. Frequent and close contact with animals or their products was significantly associated with seropositivity. No association was found with the time of employment. Sixty per cent of seropositive workers referred rheumatological symptoms. These findings confirm that slaughterhouse workers exposed to potential sources of C. jejuni have elevated titers of antibodies. Attention has, therefore, to be focused on breaking the chain of transmission as a means of control.

  7. Antimicrobial and Virulence-Modulating Effects of Clove Essential Oil on the Foodborne Pathogen Campylobacter jejuni

    PubMed Central

    Kovács, Judit K.; Felső, Péter; Makszin, Lilla; Pápai, Zoltán; Horváth, Györgyi; Ábrahám, Hajnalka; Palkovics, Tamás; Böszörményi, Andrea; Emődy, Levente

    2016-01-01

    ABSTRACT Our study investigated the antimicrobial action of clove (Syzygium aromaticum) essential oil (EO) on the zoonotic pathogen Campylobacter jejuni. After confirming the clove essential oil's general antibacterial effect, we analyzed the reference strain Campylobacter jejuni NCTC 11168. Phenotypic, proteomic, and transcriptomic methods were used to reveal changes in cell morphology and functions when exposed to sublethal concentrations of clove EO. The normally curved cells showed markedly straightened and shrunken morphology on the scanning electron micrographs as a result of stress. Although, oxidative stress, as a generally accepted response to essential oils, was also present, the dominance of a general stress response was demonstrated by reverse transcription-PCR (RT-PCR). The results of RT-PCR and two-dimensional (2D) PAGE revealed that clove oil perturbs the expression of virulence-associated genes taking part in the synthesis of flagella, PEB1, PEB4, lipopolysaccharide (LPS), and serine protease. Loss of motility was also detected by a phenotypic test. Bioautographic analysis revealed that besides its major component, eugenol, at least four other spots of clove EO possessed bactericidal activity against C. jejuni. Our findings show that clove EO has a marked antibacterial and potential virulence-modulating effect on C. jejuni. IMPORTANCE This study demonstrates that the components of clove essential oil influence not only the expression of general stress genes but also the expression of virulence-associated genes. Based on this finding, alternative strategies can be worked on to control this important foodborne pathogen. PMID:27520816

  8. Draft Genome Sequence of Campylobacter jejuni 11168H

    PubMed Central

    Macdonald, Sarah E.; Gundogdu, Ozan; Dorrell, Nick; Wren, Brendan W.; Blake, Damer

    2017-01-01

    ABSTRACT Campylobacter jejuni is the most prevalent cause of food-borne gastroenteritis in the developed world. The reference and original sequenced strain C. jejuni NCTC11168 has low levels of motility compared to clinical isolates. Here, we describe the draft genome of the laboratory derived hypermotile variant named 11168H. PMID:28153902

  9. Survival and Risk Comparison of Campylobacter jejuni on Various Processed Meat Products

    PubMed Central

    Hong, Soo Hyeon; Kim, Han Sol; Yoon, Ki Sun

    2016-01-01

    The objective of this study was to investigate survival kinetics of Campylobacter jejuni on various processed meat products (dry-cured ham, round ham with/without sodium nitrite, garlic seasoned ham with/without sodium nitrite, and sausage without sodium nitrite). Additionally, a semi-quantitative risk assessment of C. jejuni on various processed meat products was conducted using FDA-iRISK 1.0. Inoculated processed meat products with 6.0 ± 0.5 log CFU/g of C. jejuni were vacuum packed and stored at 4, 10, 17, 24, 30, and 36 °C. Survival curves were fitted to the Weibull model to obtain the delta values of C. jejuni on various processed meat products. The most rapid death of C. jejuni was observed on dry-cured ham, followed by sausage without sodium nitrite. The results of semi-quantitative risk assessment indicate that dry-cured ham represented the lowest risk among all samples. C. jejuni on processed meats presented a greater risk at 4 °C than at 10 °C. The risk of ham was greater than the risk of sausage, regardless of type. Among all samples, the highest risk of C. jejuni was observed in round ham without sodium nitrite. Overall, our data indicates that risk of processed meat products due to C. jejuni is relatively low. PMID:27294947

  10. Surface proteome mining for identification of potential vaccine candidates against Campylobacter jejuni: an in silico approach.

    PubMed

    Mehla, Kusum; Ramana, Jayashree

    2017-01-01

    Campylobacter jejuni remains a major cause of human gastroenteritis with estimated annual incidence rate of 450 million infections worldwide. C. jejuni is a major burden to public health in both socioeconomically developing and industrialized nations. Virulence determinants involved in C. jejuni pathogenesis are multifactorial in nature and not yet fully understood. Despite the completion of the first C. jejuni genome project in 2000, there are currently no vaccines in the market against this pathogen. Traditional vaccinology approach is an arduous and time extensive task. Omics techniques coupled with sequencing data have engaged researcher's attention to reduce the time and resources applied in the process of vaccine development. Recently, there has been remarkable increase in development of in silico analysis tools for efficiently mining biological information obscured in the genome. In silico approaches have been crucial for combating infectious diseases by accelerating the pace of vaccine development. This study employed a range of bioinformatics approaches for proteome scale identification of peptide vaccine candidates. Whole proteome of C. jejuni was investigated for varied properties like antigenicity, allergenicity, major histocompatibility class (MHC)-peptide interaction, immune cell processivity, HLA distribution, conservancy, and population coverage. Predicted epitopes were further tested for binding in MHC groove using computational docking studies. The predicted epitopes were conserved; covered more than 80 % of the world population and were presented by MHC-I supertypes. We conclude by underscoring that the epitopes predicted are believed to expedite the development of successful vaccines to control or prevent C. jejuni infections albeit the results need to be experimentally validated.

  11. Genomic insights from whole genome sequencing of four clonal outbreak Campylobacter jejuni assessed within the global C. jejuni population.

    PubMed

    Clark, Clifford G; Berry, Chrystal; Walker, Matthew; Petkau, Aaron; Barker, Dillon O R; Guan, Cai; Reimer, Aleisha; Taboada, Eduardo N

    2016-12-03

    Whole genome sequencing (WGS) is useful for determining clusters of human cases, investigating outbreaks, and defining the population genetics of bacteria. It also provides information about other aspects of bacterial biology, including classical typing results, virulence, and adaptive strategies of the organism. Cell culture invasion and protein expression patterns of four related multilocus sequence type 21 (ST21) C. jejuni isolates from a significant Canadian water-borne outbreak were previously associated with the presence of a CJIE1 prophage. Whole genome sequencing was used to examine the genetic diversity among these isolates and confirm that previous observations could be attributed to differential prophage carriage. Moreover, we sought to determine the presence of genome sequences that could be used as surrogate markers to delineate outbreak-associated isolates. Differential carriage of the CJIE1 prophage was identified as the major genetic difference among the four outbreak isolates. High quality single-nucleotide variant (hqSNV) and core genome multilocus sequence typing (cgMLST) clustered these isolates within expanded datasets consisting of additional C. jejuni strains. The number and location of homopolymeric tract regions was identical in all four outbreak isolates but differed from all other C. jejuni examined. Comparative genomics and PCR amplification enabled the identification of large chromosomal inversions of approximately 93 kb and 388 kb within the outbreak isolates associated with transducer-like proteins containing long nucleotide repeat sequences. The 93-kb inversion was characteristic of the outbreak-associated isolates, and the gene content of this inverted region displayed high synteny with the reference strain. The four outbreak isolates were clonally derived and differed mainly in the presence of the CJIE1 prophage, validating earlier findings linking the prophage to phenotypic differences in virulence assays and protein expression

  12. Genetic diversity in Campylobacter jejuni is associated with differential colonization of broiler chickens and C57BL/6J IL10-deficient mice

    PubMed Central

    Wilson, David L.; Rathinam, Vijay A. K.; Qi, Weihong; Wick, Lukas M.; Landgraf, Jeff; Bell, Julia A.; Plovanich-Jones, Anne; Parrish, Jodi; Finley, Russell L.; Mansfield, Linda S.; Linz, John E.

    2010-01-01

    Previous studies have demonstrated that Campylobacter jejuni, the leading causative agent of bacterial food-borne disease in the USA, exhibits high-frequency genetic variation that is associated with changes in cell-surface antigens and ability to colonize chickens. To expand our understanding of the role of genetic diversity in the disease process, we analysed the ability of three C. jejuni human disease isolates (strains 11168, 33292 and 81-176) and genetically marked derivatives to colonize Ross 308 broilers and C57BL/6J IL10-deficient mice. C. jejuni colonized broilers at much higher efficiency (all three strains, 23 of 24 broilers) than mice (11168 only, 8 of 24 mice). C. jejuni 11168 genetically marked strains colonized mice at very low efficiency (2 of 42 mice); however, C. jejuni reisolated from mice colonized both mice and broilers at high efficiency, suggesting that this pathogen can adapt genetically in the mouse. We compared the genome composition in the three wild-type C. jejuni strains and derivatives by microarray DNA/DNA hybridization analysis; the data demonstrated a high degree of genetic diversity in three gene clusters associated with synthesis and modification of the cell-surface structures capsule, flagella and lipo-oligosaccharide. Finally, we analysed the frequency of mutation in homopolymeric tracts associated with the contingency genes wlaN (GC tract) and flgR (AT tracts) in culture and after passage through broilers and mice. C. jejuni adapted genetically in culture at high frequency and the degree of genetic diversity was increased by passage through broilers but was nearly eliminated in the gastrointestinal tract of mice. The data suggest that the broiler gastrointestinal tract provides an environment which promotes outgrowth and genetic variation in C. jejuni; the enhancement of genetic diversity at this location may contribute to its importance as a human disease reservoir. PMID:20360176

  13. Discrimination of multilocus sequence typing-based Campylobacter jejuni subgroups by MALDI-TOF mass spectrometry.

    PubMed

    Zautner, Andreas Erich; Masanta, Wycliffe Omurwa; Tareen, Abdul Malik; Weig, Michael; Lugert, Raimond; Groß, Uwe; Bader, Oliver

    2013-11-07

    Campylobacter jejuni, the most common bacterial pathogen causing gastroenteritis, shows a wide genetic diversity. Previously, we demonstrated by the combination of multi locus sequence typing (MLST)-based UPGMA-clustering and analysis of 16 genetic markers that twelve different C. jejuni subgroups can be distinguished. Among these are two prominent subgroups. The first subgroup contains the majority of hyperinvasive strains and is characterized by a dimeric form of the chemotaxis-receptor Tlp7(m+c). The second has an extended amino acid metabolism and is characterized by the presence of a periplasmic asparaginase (ansB) and gamma-glutamyl-transpeptidase (ggt). Phyloproteomic principal component analysis (PCA) hierarchical clustering of MALDI-TOF based intact cell mass spectrometry (ICMS) spectra was able to group particular C. jejuni subgroups of phylogenetic related isolates in distinct clusters. Especially the aforementioned Tlp7(m+c)(+) and ansB+/ ggt+ subgroups could be discriminated by PCA. Overlay of ICMS spectra of all isolates led to the identification of characteristic biomarker ions for these specific C. jejuni subgroups. Thus, mass peak shifts can be used to identify the C. jejuni subgroup with an extended amino acid metabolism. Although the PCA hierarchical clustering of ICMS-spectra groups the tested isolates into a different order as compared to MLST-based UPGMA-clustering, the isolates of the indicator-groups form predominantly coherent clusters. These clusters reflect phenotypic aspects better than phylogenetic clustering, indicating that the genes corresponding to the biomarker ions are phylogenetically coupled to the tested marker genes. Thus, PCA clustering could be an additional tool for analyzing the relatedness of bacterial isolates.

  14. Evidence of Campylobacter jejuni reduction in broilers with early synbiotic administration.

    PubMed

    Baffoni, Loredana; Gaggìa, Francesca; Garofolo, Giuliano; Di Serafino, Gabriella; Buglione, Enrico; Di Giannatale, Elisabetta; Di Gioia, Diana

    2017-06-19

    C. jejuni is considered a food safety concern to both public health authorities and consumers since it is the leading bacterial cause of food-borne gastroenteritis in humans. A high incidence of C. jejuni in broiler flocks is often correlated to pathogen recovery in retail poultry meat, which is the main source of human infection. In this work broiler chickens were fed with a synbiotic product mixed with conventional feed using two different administration strategies. The synbiotic was formulated with the microencapsulated probiotic Bifidobacterium longum PCB133 and a xylo-oligosaccharide (XOS). 1-day old chicks were infected with C. jejuni strain M1 (10 5 cells) and the synbiotic mixture was then administered starting from the first and the 14th day of chicken life (for animal groups GrpC and GrpB respectively). The goal of this study was to monitor C. jejuni load at caecum level at different sampling time by real-time PCR, identifying the best administration strategy. The microbiological analysis of the caecal content also considered the quantification of Campylobacter spp., Bifidobacterium spp. and B. longum. The supplemented synbiotic was more successful in reducing C. jejuni and Campylobacter spp. when administered lifelong, compared to the shorter supplementation (GrpB). Bifidobacterium spp. quantification did not show significant differences among treatments and B. longum PCB133 was detected in both supplemented groups evidencing the successful colonization of the strain. Moreover, the samples of the control group (GrpA) and GrpC were analysed with PCR-denaturing gradient gel electrophoresis (PCR-DGGE) to compare the caecal microbial community profiles at the beginning and at the end of the trial. Pattern analysis evidenced the strong influence of the early synbiotic supplementation, although a physiological change in the microbial community, occurring during growth, could be observed. Experimental results demonstrate that the synbiotic approach at farm

  15. Prevalence of Campylobacter jejuni in eggs and poultry meat in New York State.

    PubMed

    Baker, R C; Paredes, M D; Qureshi, R A

    1987-11-01

    The presence of Campylobacter jejuni was tested for but not isolated from any of 276 eggs sampled from 23 egg farms in New York State. The presence of C. jejuni was evaluated in broilers, kosher broilers, spent layers, Peking ducks, and turkeys. Four of five poultry dressing plants tested showed positive growth of C. jejuni on the 25-carcass samples at various stages of processing. Twenty to 100% of live birds sampled contained C. jejuni on the skin but 90 to 100% were contaminated after scalding and defeathering operations from contaminated birds and equipment. A three to four-fold increase in carcass contamination was observed after evisceration. The number of C. jejuni on the carcasses decreased after washing and chilling. The organisms did not survive the salting, rinsing, and chilling operations in a kosher processing plant. Several pieces of equipment, i.e., shackles, eviscerating troughs, and cooling tanks were contaminated with C. jejuni. This study illustrates how C. jejuni may be transmitted from the live bird to the final poultry product.

  16. Antimicrobial and Virulence-Modulating Effects of Clove Essential Oil on the Foodborne Pathogen Campylobacter jejuni.

    PubMed

    Kovács, Judit K; Felső, Péter; Makszin, Lilla; Pápai, Zoltán; Horváth, Györgyi; Ábrahám, Hajnalka; Palkovics, Tamás; Böszörményi, Andrea; Emődy, Levente; Schneider, György

    2016-10-15

    Our study investigated the antimicrobial action of clove (Syzygium aromaticum) essential oil (EO) on the zoonotic pathogen Campylobacter jejuni After confirming the clove essential oil's general antibacterial effect, we analyzed the reference strain Campylobacter jejuni NCTC 11168. Phenotypic, proteomic, and transcriptomic methods were used to reveal changes in cell morphology and functions when exposed to sublethal concentrations of clove EO. The normally curved cells showed markedly straightened and shrunken morphology on the scanning electron micrographs as a result of stress. Although, oxidative stress, as a generally accepted response to essential oils, was also present, the dominance of a general stress response was demonstrated by reverse transcription-PCR (RT-PCR). The results of RT-PCR and two-dimensional (2D) PAGE revealed that clove oil perturbs the expression of virulence-associated genes taking part in the synthesis of flagella, PEB1, PEB4, lipopolysaccharide (LPS), and serine protease. Loss of motility was also detected by a phenotypic test. Bioautographic analysis revealed that besides its major component, eugenol, at least four other spots of clove EO possessed bactericidal activity against C. jejuni Our findings show that clove EO has a marked antibacterial and potential virulence-modulating effect on C. jejuni IMPORTANCE: This study demonstrates that the components of clove essential oil influence not only the expression of general stress genes but also the expression of virulence-associated genes. Based on this finding, alternative strategies can be worked on to control this important foodborne pathogen. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

  17. Prevention of biofilm formation and removal of existing biofilms by extracellular DNases of Campylobacter jejuni.

    PubMed

    Brown, Helen L; Reuter, Mark; Hanman, Kate; Betts, Roy P; van Vliet, Arnoud H M

    2015-01-01

    The fastidious nature of the foodborne bacterial pathogen Campylobacter jejuni contrasts with its ability to survive in the food chain. The formation of biofilms, or the integration into existing biofilms by C. jejuni, is thought to contribute to food chain survival. As extracellular DNA (eDNA) has previously been proposed to play a role in C. jejuni biofilms, we have investigated the role of extracellular DNases (eDNases) produced by C. jejuni in biofilm formation. A search of 2791 C. jejuni genomes highlighted that almost half of C. jejuni genomes contains at least one eDNase gene, but only a minority of isolates contains two or three of these eDNase genes, such as C. jejuni strain RM1221 which contains the cje0256, cje0566 and cje1441 eDNase genes. Strain RM1221 did not form biofilms, whereas the eDNase-negative strains NCTC 11168 and 81116 did. Incubation of pre-formed biofilms of NCTC 11168 with live C. jejuni RM1221 or with spent medium from a RM1221 culture resulted in removal of the biofilm. Inactivation of the cje1441 eDNase gene in strain RM1221 restored biofilm formation, and made the mutant unable to degrade biofilms of strain NCTC 11168. Finally, C. jejuni strain RM1221 was able to degrade genomic DNA from C. jejuni NCTC 11168, 81116 and RM1221, whereas strain NCTC 11168 and the RM1221 cje1441 mutant were unable to do so. This was mirrored by an absence of eDNA in overnight cultures of C. jejuni RM1221. This suggests that the activity of eDNases in C. jejuni affects biofilm formation and is not conducive to a biofilm lifestyle. These eDNases do however have a potential role in controlling biofilm formation by C. jejuni strains in food chain relevant environments.

  18. Absence of Nucleotide-Oligomerization-Domain-2 Is Associated with Less Distinct Disease in Campylobacter jejuni Infected Secondary Abiotic IL-10 Deficient Mice.

    PubMed

    Heimesaat, Markus M; Grundmann, Ursula; Alutis, Marie E; Fischer, André; Bereswill, Stefan

    2017-01-01

    Human Campylobacter jejuni -infections are progressively increasing worldwide. Despite their high prevalence and socioeconomic impact the underlying mechanisms of pathogen-host-interactions are only incompletely understood. Given that the innate immune receptor nucleotide-oligomerization-domain-2 (Nod2) is involved in clearance of enteropathogens, we here evaluated its role in murine campylobacteriosis. To address this, we applied Nod2-deficient IL-10 -/- (Nod2 -/- IL-10 -/- ) mice and IL-10 -/- counterparts both with a depleted intestinal microbiota to warrant pathogen-induced enterocolitis. At day 7 following peroral C. jejuni strain 81-176 infection, Nod2 mRNA was down-regulated in the colon of secondary abiotic IL-10 -/- and wildtype mice. Nod2-deficiency did neither affect gastrointestinal colonization nor extra-intestinal and systemic translocation properties of C. jejuni . Colonic mucin-2 mRNA was, however, down-regulated upon C. jejuni -infection of both Nod2 -/- IL-10 -/- and IL-10 -/- mice, whereas expression levels were lower in infected, but also naive Nod2 -/- IL-10 -/- mice as compared to respective IL-10 -/- controls. Remarkably, C. jejuni -infected Nod2 -/- IL-10 -/- mice were less compromised than IL-10 -/- counterparts and displayed less distinct apoptotic, but higher regenerative cell responses in colonic epithelia. Conversely, innate as well as adaptive immune cells such as macrophages and monocytes as well as T lymphocytes and regulatory T-cells, respectively, were even more abundant in large intestines of Nod2 -/- IL-10 -/- as compared to IL-10 -/- mice at day 7 post-infection. Furthermore, IFN-γ concentrations were higher in ex vivo biopsies derived from intestinal compartments including colon and mesenteric lymph nodes as well as in systemic tissue sites such as the spleen of C. jejuni infected Nod2 -/- IL-10 -/- as compared to IL10 -/- counterparts. Whereas, at day 7 postinfection anti-inflammatory IL-22 mRNA levels were up-regulated, IL

  19. Complete genome sequences of three Campylobacter jejuni phage-propagating strains

    USDA-ARS?s Scientific Manuscript database

    Bacteriophage therapy has the potential to reduce Campylobacter jejuni numbers in livestock, but requires a detailed understanding of phage-host interactions. Some C. jejuni strains are readily infected by certain phages, and are thus designated as phage-propagating strains. Here we report the compl...

  20. The RNase R from Campylobacter jejuni Has Unique Features and Is Involved in the First Steps of Infection*

    PubMed Central

    Haddad, Nabila; Matos, Rute G.; Pinto, Teresa; Rannou, Pauline; Cappelier, Jean-Michel; Prévost, Hervé; Arraiano, Cecília M.

    2014-01-01

    Bacterial pathogens must adapt/respond rapidly to changing environmental conditions. Ribonucleases (RNases) can be crucial factors contributing to the fast adaptation of RNA levels to different environmental demands. It has been demonstrated that the exoribonuclease polynucleotide phosphorylase (PNPase) facilitates survival of Campylobacter jejuni in low temperatures and favors swimming, chick colonization, and cell adhesion/invasion. However, little is known about the mechanism of action of other ribonucleases in this microorganism. Members of the RNB family of enzymes have been shown to be involved in virulence of several pathogens. We have searched C. jejuni genome for homologues and found one candidate that displayed properties more similar to RNase R (Cj-RNR). We show here that Cj-RNR is important for the first steps of infection, the adhesion and invasion of C. jejuni to eukaryotic cells. Moreover, Cj-RNR proved to be active in a wide range of conditions. The results obtained lead us to conclude that Cj-RNR has an important role in the biology of this foodborne pathogen. PMID:25100732

  1. The food-borne pathogen Campylobacter jejuni depends on the AddAB DNA repair system to defend against bile in the intestinal environment.

    PubMed

    Gourley, Christopher R; Negretti, Nicholas M; Konkel, Michael E

    2017-10-31

    Accurate repair of DNA damage is crucial to ensure genome stability and cell survival of all organisms. Bile functions as a defensive barrier against intestinal colonization by pathogenic microbes. Campylobacter jejuni, a leading bacterial cause of foodborne illness, possess strategies to mitigate the toxic components of bile. We recently found that growth of C. jejuni in medium with deoxycholate, a component of bile, caused DNA damage consistent with the exposure to reactive oxygen species. We hypothesized that C. jejuni must repair DNA damage caused by reactive oxygen species to restore chromosomal integrity. Our efforts focused on determining the importance of the putative AddAB DNA repair proteins. A C. jejuni addAB mutant demonstrated enhanced sensitivity to deoxycholate and was impaired in DNA double strand break repair. Complementation of the addAB mutant restored resistance to deoxycholate, as well as function of the DNA double strand break repair system. The importance of these findings translated to the natural host, where the AddAB system was found to be required for efficient C. jejuni colonization of the chicken intestine. This research provides new insight into the molecular mechanism utilized by C. jejuni, and possibly other intestinal pathogens, to survive in the presence of bile.

  2. L-fucose utilization provides Campylobacter jejuni with a competitive advantage.

    PubMed

    Stahl, Martin; Friis, Lorna M; Nothaft, Harald; Liu, Xin; Li, Jianjun; Szymanski, Christine M; Stintzi, Alain

    2011-04-26

    Campylobacter jejuni is a prevalent gastrointestinal pathogen in humans and a common commensal of poultry. When colonizing its hosts, C. jejuni comes into contact with intestinal carbohydrates, including L-fucose, released from mucin glycoproteins. Several strains of C. jejuni possess a genomic island (cj0480c-cj0490) that is up-regulated in the presence of both L-fucose and mucin and allows for the utilization of L-fucose as a substrate for growth. Strains possessing this genomic island show increased growth in the presence of L-fucose and mutation of cj0481, cj0486, and cj0487 results in the loss of the ability to grow on this substrate. Furthermore, mutants in the putative fucose permease (cj0486) are deficient in fucose uptake and demonstrate a competitive disadvantage when colonizing the piglet model of human disease, which is not paralleled in the colonization of poultry. This identifies a previously unrecorded metabolic pathway in select strains of C. jejuni associated with a virulent lifestyle.

  3. l-Fucose utilization provides Campylobacter jejuni with a competitive advantage

    PubMed Central

    Stahl, Martin; Friis, Lorna M.; Nothaft, Harald; Liu, Xin; Li, Jianjun; Szymanski, Christine M.; Stintzi, Alain

    2011-01-01

    Campylobacter jejuni is a prevalent gastrointestinal pathogen in humans and a common commensal of poultry. When colonizing its hosts, C. jejuni comes into contact with intestinal carbohydrates, including l-fucose, released from mucin glycoproteins. Several strains of C. jejuni possess a genomic island (cj0480c–cj0490) that is up-regulated in the presence of both l-fucose and mucin and allows for the utilization of l-fucose as a substrate for growth. Strains possessing this genomic island show increased growth in the presence of l-fucose and mutation of cj0481, cj0486, and cj0487 results in the loss of the ability to grow on this substrate. Furthermore, mutants in the putative fucose permease (cj0486) are deficient in fucose uptake and demonstrate a competitive disadvantage when colonizing the piglet model of human disease, which is not paralleled in the colonization of poultry. This identifies a previously unrecorded metabolic pathway in select strains of C. jejuni associated with a virulent lifestyle. PMID:21482772

  4. In-water supplementation of Trans-cinnamaldehyde nanoemulsion reduces Campylobacter jejuni colonization in broiler chickens

    USDA-ARS?s Scientific Manuscript database

    Campylobacter jejuni is a major foodborne pathogen that causes severe gastroenteritis in humans. Chickens act as the reservoir host for C. jejuni, wherein the pathogen colonizes the ceca thereby leading to contamination of the carcass during slaughter. Reducing C. jejuni cecal colonization could pot...

  5. Anti-adhesion activity of thyme (Thymus vulgaris L.) extract, thyme post-distillation waste, and olive (Olea europea L.) leaf extract against Campylobacter jejuni on polystyrene and intestine epithelial cells.

    PubMed

    Šikić Pogačar, Maja; Klančnik, Anja; Bucar, Franz; Langerholc, Tomaž; Smole Možina, Sonja

    2016-06-01

    In order to survive in food-processing environments and cause disease, Campylobacter jejuni requires specific survival mechanisms, such as biofilms, which contribute to its transmission through the food chain to the human host and present a critical form of resistance to a wide variety of antimicrobials. Phytochemical analysis of thyme ethanolic extract (TE), thyme post-hydrodistillation residue (TE-R), and olive leaf extract (OE) using high-performance liquid chromatography with photodiode array indicates that the major compounds in TE and TE-R are flavone glucuronides and rosmarinic acid derivatives, and in OE verbascoside, luteolin 7-O-glucoside and oleuroside. TE and TE-R reduced C. jejuni adhesion to abiotic surfaces by up to 30% at 0.2-12.5 µg mL(-1) , with TE-R showing a greater effect. OE from 3.125 to 200 µg mL(-1) reduced C. jejuni adhesion to polystyrene by 10-23%. On the other hand, C. jejuni adhesion to PSI cl1 cells was inhibited by almost 30% over a large concentration range of these extracts. Our findings suggest that TE, the agro-food waste material TE-R, and the by-product OE represent sources of bioactive phytochemicals that are effective at low concentrations and can be used as therapeutic agents to prevent bacterial adhesion. © 2015 Society of Chemical Industry. © 2015 Society of Chemical Industry.

  6. [Experimental study on the chitosan-DNA vaccines against campylobacter jejuni invasion].

    PubMed

    Zheng, Hui; Cai, Fang-cheng; Zhong, Min; Deng, Bing; Li, Xin; Zhang, Xiao-ping

    2007-09-01

    The immunogenicity and protective efficacy of an experimental Campylobacter jejuni (C. jejuni) chitosan-DNA vaccines were evaluated in mice. The chitosan-DNA vaccines were prepared by embedding pcDNA3.1(+)-cadF and pcDNA3.1(+)-peblA with chitosan respectively. BALB/c mice were intranasally immunized in a four-dose primary series (7 d intervals) at doses of 60 microg chitosan-DNA vaccines each time. The comparative immunogenicities of nine formulations were assessed on the basis of the generation of antigen-specific antibodies in serum and intestinal secretions. Mice were attacked repeatedly through intragastric administration of C. jejuni HS:19 at the 8th week after the immunization and protective efficacy was determined by detecting the degrees of protection afforded against C. jejuni invaded. The mice immunized with chitosan-DNA vaccines have generated high levels of IgA and IgG from the sera and IgA from the intestinal secretions and the P/N value went up to 20.58, 30.13 and 6.87 respectively. Meanwhile, the expression of intestinal SIgA increased correspondingly. Moreover the chitosan-DNA vaccines induced strongest level of protection in BALB/c mice against challenge with C. jejuni HS:19 strain and the protective efficacies was 93.70. The results of this study indicate that the chitosan-DNA vaccines could induce significant protective immunity against C. jejuni challenge in the mice model.

  7. γ-Glutamyl transpeptidase has a role in the persistent colonization of the avian gut by Campylobacter jejuni

    PubMed Central

    Barnes, If H.A.; Bagnall, Mary C.; Browning, Darren D.; Thompson, Stuart A.; Manning, Georgina; Newell, Diane G.

    2009-01-01

    The contribution of γ-glutamyl transpeptidase (GGT) to Campylobacter jejuni virulence and colonization of the avian gut has been investigated. The presence of the ggt gene in C. jejuni strains directly correlated with the expression of GGT activity as measured by cleavage and transfer of the γ-glutamyl moiety. Inactivation of the monocistronic ggt gene in C. jejuni strain 81116 resulted in isogenic mutants with undetectable GGT activity; nevertheless, these mutants grew normally in vitro. However, the mutants had increased motility, a 5.4-fold higher invasion efficiency into INT407 cells in vitro and increased resistance to hydrogen peroxide stress. Moreover, the apoptosis-inducing activity of the ggt mutant was significantly lower than that of the parental strain. In vivo studies showed that, although GGT activity was not required for initial colonization of 1-day-old chicks, the enzyme was required for persistant colonization of the avian gut. PMID:17600669

  8. [Campylobacter jejuni in poultry processed in slaughterhouses].

    PubMed

    Mícková, V

    1987-09-01

    The frequency of occurrence of Campylobacter jejuni germs in dressed poultry was studied for a year. The samples--smears from the body cavities of chickens--were collected during the technological dressing of the chickens; 101 strains of Campylobacter jejuni (i. e. 28.69%) were isolated from the 352 samples analyzed. The occurrence of the germs exhibited a considerable seasonal variance with peak rates in spring and summer. The use of a suitable culture medium, the technique of cultivation and the properties of the isolated strains were studied at the same time. The culture medium (Agar no. 3 IMUNA enriched with supplement C, horse blood and ingredients increasing the aerotolerance of the germs--sodium pyruvate and iron sulphate) used during the investigation was found to be suitable. The technique of cultivation by means of an anaerostat manufactured by the Development Station in Brno, atmosphere regulation (5% CO2) and with a pre-set cultivation temperature (43 degrees C) was found to be suitable for the screening of the Campylobacter jejuni germs.

  9. Campylobacter jejuni survival in a poultry processing plant environment.

    PubMed

    García-Sánchez, Lourdes; Melero, Beatriz; Jaime, Isabel; Hänninen, Marja-Liisa; Rossi, Mirko; Rovira, Jordi

    2017-08-01

    Campylobacteriosis is the most common cause of bacterial gastroenteritis worldwide. Consumption of poultry, especially chicken's meat is considered the most common route for human infection. The aim of this study was to determine if Campylobacter spp. might persist in the poultry plant environment before and after cleaning and disinfection procedures and the distribution and their genetic relatedness. During one month from a poultry plant were analyzed a total of 494 samples -defeathering machine, evisceration machine, floor, sink, conveyor belt, shackles and broiler meat- in order to isolate C. jejuni and C. coli. Results showed that C. jejuni and C. coli prevalence was 94.5% and 5.5% respectively. Different typing techniques as PFGE, MLST established seven C. jejuni genotypes. Whole genome MLST strongly suggest that highly clonal populations of C. jejuni can survive in adverse environmental conditions, even after cleaning and disinfection, and persist for longer periods than previous thought (at least 21 days) in the poultry plant environment. Even so, it might act as a source of contamination independently of the contamination level of the flock entering the slaughter line. Copyright © 2017 Elsevier Ltd. All rights reserved.

  10. Chicken Caecal Microbiome Modifications Induced by Campylobacter jejuni Colonization and by a Non-Antibiotic Feed Additive.

    PubMed

    Thibodeau, Alexandre; Fravalo, Philippe; Yergeau, Étienne; Arsenault, Julie; Lahaye, Ludovic; Letellier, Ann

    2015-01-01

    Campylobacter jejuni is an important zoonotic foodborne pathogen causing acute gastroenteritis in humans. Chickens are often colonized at very high numbers by C. jejuni, up to 10(9) CFU per gram of caecal content, with no detrimental effects on their health. Farm control strategies are being developed to lower the C. jejuni contamination of chicken food products in an effort to reduce human campylobacteriosis incidence. It is believed that intestinal microbiome composition may affect gut colonization by such undesirable bacteria but, although the chicken microbiome is being increasingly characterized, information is lacking on the factors affecting its modulation, especially by foodborne pathogens. This study monitored the effects of C. jejuni chicken caecal colonization on the chicken microbiome in healthy chickens. It also evaluated the capacity of a feed additive to affect caecal bacterial populations and to lower C. jejuni colonization. From day-0, chickens received or not a microencapsulated feed additive and were inoculated or not with C. jejuni at 14 days of age. Fresh caecal content was harvested at 35 days of age. The caecal microbiome was characterized by real time quantitative PCR and Ion Torrent sequencing. We observed that the feed additive lowered C. jejuni caecal count by 0.7 log (p<0.05). Alpha-diversity of the caecal microbiome was not affected by C. jejuni colonization or by the feed additive. C. jejuni colonization modified the caecal beta-diversity while the feed additive did not. We observed that C. jejuni colonization was associated with an increase of Bifidobacterium and affected Clostridia and Mollicutes relative abundances. The feed additive was associated with a lower Streptococcus relative abundance. The caecal microbiome remained relatively unchanged despite high C. jejuni colonization. The feed additive was efficient in lowering C. jejuni colonization while not disturbing the caecal microbiome.

  11. Chicken Caecal Microbiome Modifications Induced by Campylobacter jejuni Colonization and by a Non-Antibiotic Feed Additive

    PubMed Central

    Thibodeau, Alexandre; Fravalo, Philippe; Yergeau, Étienne; Arsenault, Julie; Lahaye, Ludovic; Letellier, Ann

    2015-01-01

    Campylobacter jejuni is an important zoonotic foodborne pathogen causing acute gastroenteritis in humans. Chickens are often colonized at very high numbers by C. jejuni, up to 109 CFU per gram of caecal content, with no detrimental effects on their health. Farm control strategies are being developed to lower the C. jejuni contamination of chicken food products in an effort to reduce human campylobacteriosis incidence. It is believed that intestinal microbiome composition may affect gut colonization by such undesirable bacteria but, although the chicken microbiome is being increasingly characterized, information is lacking on the factors affecting its modulation, especially by foodborne pathogens. This study monitored the effects of C. jejuni chicken caecal colonization on the chicken microbiome in healthy chickens. It also evaluated the capacity of a feed additive to affect caecal bacterial populations and to lower C. jejuni colonization. From day-0, chickens received or not a microencapsulated feed additive and were inoculated or not with C. jejuni at 14 days of age. Fresh caecal content was harvested at 35 days of age. The caecal microbiome was characterized by real time quantitative PCR and Ion Torrent sequencing. We observed that the feed additive lowered C. jejuni caecal count by 0.7 log (p<0.05). Alpha-diversity of the caecal microbiome was not affected by C. jejuni colonization or by the feed additive. C. jejuni colonization modified the caecal beta-diversity while the feed additive did not. We observed that C. jejuni colonization was associated with an increase of Bifidobacterium and affected Clostridia and Mollicutes relative abundances. The feed additive was associated with a lower Streptococcus relative abundance. The caecal microbiome remained relatively unchanged despite high C. jejuni colonization. The feed additive was efficient in lowering C. jejuni colonization while not disturbing the caecal microbiome. PMID:26161743

  12. High frequency, spontaneous motA mutations in Campylobacter jejuni strain 81-176.

    PubMed

    Mohawk, Krystle L; Poly, Frédéric; Sahl, Jason W; Rasko, David A; Guerry, Patricia

    2014-01-01

    Campylobacter jejuni is an important cause of bacterial diarrhea worldwide. The pathogenesis of C. jejuni is poorly understood and complicated by phase variation of multiple surface structures including lipooligosaccharide, capsule, and flagellum. When C. jejuni strain 81-176 was plated on blood agar for single colonies, the presence of translucent, non-motile colonial variants was noted among the majority of opaque, motile colonies. High-throughput genomic sequencing of two flagellated translucent and two opaque variants as well as the parent strain revealed multiple genetic changes compared to the published genome. However, the only mutated open reading frame common between the two translucent variants and absent from the opaque variants and the parent was motA, encoding a flagellar motor protein. A total of 18 spontaneous motA mutations were found that mapped to four distinct sites in the gene, with only one class of mutation present in a phase variable region. This study exemplifies the mutative/adaptive properties of C. jejuni and demonstrates additional variability in C. jejuni beyond phase variation.

  13. Phenotypic and Transcriptomic Responses of Campylobacter jejuni Suspended in an Artificial Freshwater Medium

    PubMed Central

    Trigui, Hana; Lee, Kristen; Thibodeau, Alexandre; Lévesque, Simon; Mendis, Nilmini; Fravalo, Philippe; Letellier, Ann; Faucher, Sébastien P.

    2017-01-01

    Campylobacter jejuni is the leading cause of campylobacteriosis in the developed world. Although most cases are caused by consumption of contaminated meat, a significant proportion is linked to ingestion of contaminated water. The differences between C. jejuni strains originating from food products and those isolated from water are poorly understood. Working under the hypothesis that water-borne C. jejuni strains are better equipped at surviving the nutrient-poor aquatic environment than food-borne strains, the present study aims to characterize these differences using outbreak strains 81116 and 81-176. Strain 81116 caused a campylobacteriosis outbreak linked to consumption of water, while strain 81-176 was linked to consumption of raw milk. CFU counts and viability assays showed that 81116 survives better than 81-176 at 4°C in a defined freshwater medium (Fraquil). Moreover, 81116 was significantly more resistant to oxidative stress and bile salt than strain 81-176 in Fraquil. To better understand the genetic response of 81116 to water, a transcriptomic profiling study was undertaken using microarrays. Compared to rich broth, strain 81116 represses genes involved in amino acid uptake and metabolism, as well as genes involved in costly biosynthetic processes such as replication, translation, flagellum synthesis and virulence in response to Fraquil. In accordance with the observed increase in stress resistance in Fraquil, 81116 induces genes involved in resistance to oxidative stress and bile salt. Interestingly, genes responsible for cell wall synthesis were also induced upon Fraquil exposure. Finally, twelve unique genes were expressed in Fraquil; however, analysis of their distribution in animal and water isolates showed that they are not uniquely and ubiquitously present in water isolates, and thus, unlikely to play a major role in adaptation to water. Our results show that some C. jejuni strains are more resilient than others, thereby challenging current water

  14. Defining the metabolic requirements for the growth and colonization capacity of Campylobacter jejuni

    PubMed Central

    Hofreuter, Dirk

    2014-01-01

    During the last decade Campylobacter jejuni has been recognized as the leading cause of bacterial gastroenteritis worldwide. This facultative intracellular pathogen is a member of the Epsilonproteobacteria and requires microaerobic atmosphere and nutrient rich media for efficient proliferation in vitro. Its catabolic capacity is highly restricted in contrast to Salmonella Typhimurium and other enteropathogenic bacteria because several common pathways for carbohydrate utilization are either missing or incomplete. Despite these metabolic limitations, C. jejuni efficiently colonizes various animal hosts as a commensal intestinal inhabitant. Moreover, C. jejuni is tremendously successful in competing with the human intestinal microbiota; an infectious dose of few hundreds bacteria is sufficient to overcome the colonization resistance of humans and can lead to campylobacteriosis. Besides the importance and clear clinical manifestation of this disease, the pathogenesis mechanisms of C. jejuni infections are still poorly understood. In recent years comparative genome sequence, transcriptome and metabolome analyses as well as mutagenesis studies combined with animal infection models have provided a new understanding of how the specific metabolic capacity of C. jejuni drives its persistence in the intestinal habitat of various hosts. Furthermore, new insights into the metabolic requirements that support the intracellular survival of C. jejuni were obtained. Because C. jejuni harbors distinct properties in establishing an infection in comparison to pathogenic Enterobacteriaceae, it represents an excellent organism for elucidating new aspects of the dynamic interaction and metabolic cross talk between a bacterial pathogen, the microbiota and the host. PMID:25325018

  15. Synergistic anti-Campylobacter jejuni activity of fluoroquinolone and macrolide antibiotics with phenolic compounds

    PubMed Central

    Oh, Euna; Jeon, Byeonghwa

    2015-01-01

    The increasing resistance of Campylobacter to clinically important antibiotics, such as fluoroquinolones and macrolides, is a serious public health problem. The objective of this study is to investigate synergistic anti-Campylobacter jejuni activity of fluoroquinolones and macrolides in combination with phenolic compounds. Synergistic antimicrobial activity was measured by performing a checkerboard assay with ciprofloxacin and erythromycin in the presence of 21 phenolic compounds. Membrane permeability changes in C. jejuni by phenolic compounds were determined by measuring the level of intracellular uptake of 1-N-phenylnaphthylamine (NPN). Antibiotic accumulation assays were performed to evaluate the level of ciprofloxacin accumulation in C. jejuni. Six phenolic compounds, including p-coumaric acid, sinapic acid, caffeic acid, vanillic acid, gallic acid, and taxifolin, significantly increased the susceptibility to ciprofloxacin and erythromycin in several human and poultry isolates. The synergistic antimicrobial effect was also observed in ciprofloxacin- and erythromycin-resistant C. jejuni strains. The phenolic compounds also substantially increased membrane permeability and antibiotic accumulation in C. jejuni. Interestingly, some phenolic compounds, such as gallic acid and taxifolin, significantly reduced the expression of the CmeABC multidrug efflux pump. Phenolic compounds increased the NPN accumulation in the cmeB mutant, indicating phenolic compounds may affect the membrane permeability. In this study, we successfully demonstrated that combinational treatment of C. jejuni with antibiotics and phenolic compounds synergistically inhibits C. jejuni by impacting both antimicrobial influx and efflux. PMID:26528273

  16. Antibiotic susceptibility profiling and virulence potential of Campylobacter jejuni isolates from different sources in Pakistan.

    PubMed

    Siddiqui, Fariha Masood; Akram, Muhammad; Noureen, Nighat; Noreen, Zobia; Bokhari, Habib

    2015-03-01

    To determine antibiotic resistance patterns and virulence potential of Campylobacter jejuni (C. jejuni) isolates from clinical human diarrheal infections, cattle and healthy broilers. Antibiotic sensitivity patterns of C. jejuni isolates were determined by Kirby Bauer Disc Diffusion assay. These isolates were then subjected to virulence profiling for the detection of mapA (membrane-associated protein), cadF (fibronectin binding protein), wlaN (beta-l,3-galactosyltransferase) and neuAB (sialic acid biosynthesis gene). Further C. jejuni isolates were grouped by random amplification of polymorphic DNA (RAPD) profiling. A total of 436 samples from poultry (n=88), cattle (n=216) and humans (n=132) from different locations were collected. Results revealed percentage of C. jejuni isolates were 35.2% (31/88), 25.0% (54/216) and 11.3% (15/132) among poultry, cattle and clinical human samples respectively. Antibiotic susceptibility results showed that similar resistance patterns to cephalothin was ie. 87.0%, 87.1% and 89%among humans, poultry and cattle respectively, followed by sulfamethoxazole+trimethoprim 40.0%, 38.7% and 31.0% in humans, poultry and cattle and Ampicillin 40%, 32% and 20% in humans, poultry and cattle respectively. Beta-lactamase activity was detected in 40.00% humans, 20.37% cattle and 32.25% in poultry C. jejuni isolates. CadF and mapA were present in all poultry, cattle and human C. jejuni isolates, wlaN was not detected in any isolate and neuAB was found in 9/31 (36%) poultry isolates. RAPD profiling results suggested high diversity of C. jejuni isolates. Detection of multidrug resistant C. jejuni strains from poultry and cattle is alarming as they can be potential hazard to humans. Moreover, predominant association of virulence factors, cadF and mapA (100% each) in C. jejuni isolates from all sources and neuAB (36%) with poultry isolates suggest the potential source of transmission of diverse types of C. jejuni to humans. Copyright © 2015 Hainan

  17. Therapeutic administration of enrofloxacin in mice does not select for fluoroquinolone resistance in Campylobacter jejuni.

    PubMed

    Inglis, G Douglas; Zaytsoff, S J M; Selinger, L Brent; Taboada, Eduardo N; Uwiera, R R E

    2018-05-11

    Enrofloxacin is registered for therapeutic use in beef cattle to treat bovine respiratory disease in Canada. A murine model was used to experimentally examine the impact of therapeutic administration of enrofloxacin on fluoroquinolone resistance development in Campylobacter jejuni. Administration of enrofloxacin to mice via subcutaneous injection or per os routes resulted in equivalent levels of bioactive enrofloxacin within the intestine, but bioactivity was short-lived (<48 hr after cessation). Enrofloxacin administration did not affect densities of total bacteria, Firmicutes, or Bacteroidetes in digesta, and had modest impacts on densities of Enterobacteriaceae. All mice inoculated with C. jejuni NCTC 11168 became persistently colonized by the bacterium. Enrofloxacin reduced C. jejuni cell densities within the cecal and colonic digesta for all treatments, and densities shed in feces as a function of antibiotic duration. None of the C. jejuni isolates recovered from mice after administration of enrofloxacin (n=260) developed resistance to ciprofloxacin regardless of method or duration of administration. Furthermore, only modest shifts in the minimum inhibitory concentration of the isolates by treatment were noted. The study findings indicate that the risk posed by short-term subcutaneous administration of enrofloxacin for the development of fluoroquinolone resistance in mammals is low.

  18. [Characterization of Campylobacter jejuni samples coming form poultry meat and feces].

    PubMed

    Gutiérrez, Sindy; Orellana, Daniel; Martínez, Claudio; García Mena, Verónica

    2017-12-01

    Background Campylobacter jejuni is one of the main causal agents of food borne diseases. Infections with this pathogen are mainly caused by chicken meat consumption. Aim To characterize antibiotic resistance and virulence factors in C. jejuni strains obtained from chicken meat and poultry feces in Central Chile. Material and Methods The presence of C. jejuni in 30 meat and 40 feces samples from poultry was studied. From these samples, we obtained 40 strains which were characterized at the molecular level for the presence of 16 genes involved in virulence using PCR. In parallel, antibiotic resistance for ciprofloxacin, nalidixic acid, tetracycline, erythromycin, azithromycin, chloramphenicol y ampicillin was analyzed. Results Twenty and 63% of feces and chicken meat samples were positive for C. jejuni, respectively. Moreover, a high percentage of strains showed antibiotic resistance, where 27% of strains were resistant to all tested antibiotics, except for azithromycin. Finally, 10% of the strains coming from feces contained 14 out of 16 virulence genes evaluated. Only 23% of the strains did not contain any of these genes. Conclusions A high percentage of feces and chicken meat samples are contaminated with C. jejuni. Moreover, these strains show a high genetic and phenotypic diversity represented by their antibiotic resistance profiles and the presence of virulence factors.

  19. Insights into potential pathogenesis mechanisms associated with Campylobacter jejuni-induced abortion in ewes.

    PubMed

    Sanad, Yasser M; Jung, Kwonil; Kashoma, Isaac; Zhang, Xiaoli; Kassem, Issmat I; Saif, Yehia M; Rajashekara, Gireesh

    2014-11-25

    Campylobacter jejuni is commonly found in the gastrointestinal tract of many food-animals including sheep without causing visible clinical symptoms of disease. However, C. jejuni has been implicated in ovine abortion cases worldwide. Specifically, in the USA, the C. jejuni sheep abortion (SA) clone has been increasingly associated with sheep abortion. In vivo studies in sheep (the natural host) are needed to better characterize the virulence potential and pathogenesis of this clone. Pregnant ewes intravenously (IV) or orally inoculated with ovine or bovine abortion-associated C. jejuni SA clones exhibited partial or complete uterine prolapse with retained placenta, and abortion or stillbirth, whereas delivery of healthy lambs occurred in pregnant ewes inoculated with C. jejuni 81-176 or in the uninfected group. In sheep inoculated with the SA clone, histopathological lesions including suppurative necrotizing placentitis and/or endometritis coincided with: 1) increased apoptotic death of trophoblasts, 2) increased expression of the host genes (e.g. genes encoding interleukin IL-6 and IL-15) related to cellular necrosis and pro-inflammatory responses in uterus, and 3) decreased expression of the genes encoding GATA binding protein 6, chordin, and insulin-like 3 (INSL3) that account for embryonic development in uterus. Immunohistochemistry revealed localization of bacterial antigens in trophoblasts lining the chorioallantoic membrane of ewes inoculated with the C. jejuni SA clone. The results showed that C. jejuni SA clones are capable of causing abortion or stillbirth in experimentally infected sheep. Furthermore, down- or up-regulation of specific genes in the uterus of infected pregnant ewes might implicate host genes in facilitating the disease progression. Since the C. jejuni SA strains share genotypic similarities with clones that have been isolated from human clinical cases of gastroenteritis, these strains might represent a potential public health risk.

  20. Detection of Campylobacter jejuni in rectal swab samples from Rousettus amplexicaudatus in the Philippines.

    PubMed

    Hatta, Yuki; Omatsu, Tsutomu; Tsuchiaka, Shinobu; Katayama, Yukie; Taniguchi, Satoshi; Masangkay, Joseph S; Puentespina, Roberto; Eres, Eduardo; Cosico, Edison; Une, Yumi; Yoshikawa, Yasuhiro; Maeda, Ken; Kyuwa, Shigeru; Mizutani, Tetsuya

    2016-09-01

    Bats are the second diversity species of mammals and widely distributed in the world. They are thought to be reservoir and vectors of zoonotic pathogens. However, there is scarce report of the evidence of pathogenic bacteria kept in bats. The precise knowledge of the pathogenic bacteria in bat microbiota is important for zoonosis control. Thus, metagenomic analysis targeting the V3-V4 region of the 16S rRNA of the rectal microbiota in Rousettus amplexicaudatus was performed using high throughput sequencing. The results revealed that 103 genera of bacteria including Camplyobacter were detected. Campylobacter was second predominant genus, and Campylobacter coli and Campylobacter jejuni were identified in microbiome of R. amplexicaudatus. Campylobacteriosis is one of the serious bacterial diarrhea in human, and the most often implicated species as the causative agent of campylobacteriosis is C. jejuni. Therefore, we investigated the prevalence of C. jejuni in 91 wild bats with PCR. As a result of PCR assay targeted on 16S-23S intergenic spacer, partial genome of C. jejuni was detected only in five R. amplexicaudatus. This is the first report that C. jejuni was detected in bat rectal swab samples. C. jejuni is the most common cause of campylobacteriosis in humans, transmitted through water and contact with livestock animals. This result indicated that R. amplexicaudatus may be a carrier of C. jejuni.

  1. Complete genome sequence of Campylobacter jejuni strain 12567 a livestock-associated clade representative

    USDA-ARS?s Scientific Manuscript database

    We report the complete genome sequence of the Campylobacter jejuni strain 12567, a member of a C. jejuni livestock-associated clade that expresses glycoconjugates linked to improved gastrointestinal tract persistence....

  2. Campylobacter jejuni: A rare agent in a child with peritoneal dialysis-related peritonitis.

    PubMed

    Tural Kara, Tugce; Yilmaz, Songul; Ozdemir, Halil; Birsin Ozcakar, Zeynep; Derya Aysev, Ahmet; Ciftci, Ergin; Ince, Erdal

    2016-10-01

    Peritonitis is a serious problem in children receiving peritoneal dialysis. Campylobacter jejuni is an unusual cause of peritonitis. A 10-year-old boy who had end stage renal failure due to atypical hemolytic uremic syndrome was admitted to our hospital with abdominal pain and fever. Peritoneal dialysis fluid was cloudy and microscopic examination showed abundant leukocytes. Intraperitoneal cefepime treatment was started. Campylobacter jejuni was isolated from peritoneal dialysis fluid culture and oral clarithromycin was added to the treatment. At the end of therapy, peritoneal fluid culture was negative. To our knowledge, C. jejuni peritonitis was not reported in children previously. Although C. jejuni peritonitis is rarely encountered in children, it should be considered as an etiologic factor for peritonitis. Sociedad Argentina de Pediatría.

  3. A DNase encoded by integrated element CJIE1 inhibits natural transformation of Campylobacter jejuni.

    PubMed

    Gaasbeek, Esther J; Wagenaar, Jaap A; Guilhabert, Magalie R; Wösten, Marc M S M; van Putten, Jos P M; van der Graaf-van Bloois, Linda; Parker, Craig T; van der Wal, Fimme J

    2009-04-01

    The species Campylobacter jejuni is considered naturally competent for DNA uptake and displays strong genetic diversity. Nevertheless, nonnaturally transformable strains and several relatively stable clonal lineages exist. In the present study, the molecular mechanism responsible for the nonnatural transformability of a subset of C. jejuni strains was investigated. Comparative genome hybridization indicated that C. jejuni Mu-like prophage integrated element 1 (CJIE1) was more abundant in nonnaturally transformable C. jejuni strains than in naturally transformable strains. Analysis of CJIE1 indicated the presence of dns (CJE0256), which is annotated as a gene encoding an extracellular DNase. DNase assays using a defined dns mutant and a dns-negative strain expressing Dns from a plasmid indicated that Dns is an endogenous DNase. The DNA-hydrolyzing activity directly correlated with the natural transformability of the knockout mutant and the dns-negative strain expressing Dns from a plasmid. Analysis of a broader set of strains indicated that the majority of nonnaturally transformable strains expressed DNase activity, while all naturally competent strains lacked this activity. The inhibition of natural transformation in C. jejuni via endogenous DNase activity may contribute to the formation of stable lineages in the C. jejuni population.

  4. Antimicrobial wash with Trans-cinnamaldehyde nanoemulsion reduces Campylobacter jejuni on chicken skin

    USDA-ARS?s Scientific Manuscript database

    Campylobacter jejuni is a major foodborne pathogen that causes severe enteritis in humans largely due to consumption of contaminated poultry products. Reducing C. jejuni contamination on chicken carcasses would reduce subsequent human infections. This study investigated the efficacy of Trans-cinnama...

  5. Reactions of chicken sera to recombinant Campylobacter jejuni flagellar proteins.

    PubMed

    Yeh, Hung-Yueh; Hiett, Kelli L; Line, John E

    2015-03-01

    Campylobacter jejuni is a Gram-negative spiral rod bacterium and is the leading but underreported bacterial food-borne pathogen that causes human campylobacteriosis worldwide. Raw or undercooked poultry products are regarded as a major source for human infection. C. jejuni flagella have been implicated in colonization and adhesion to the mucosal surface of chicken gastrointestinal tracts. Therefore, flagellar proteins would be the excellent targets for further investigation. In this report, we used the recombinant technology to generate a battery of C. jejuni flagellar proteins, which were purified by His tag affinity chromatography and determined antigenic profiles of these recombinant flagellar proteins using sera from chickens older than 6 weeks of age. The immunoblot results demonstrate that each chicken serum reacted to various numbers of recombinant flagellar proteins. Among these recombinant proteins, chicken sera reacted predominantly to the FlgE1, FlgK, FlhF, FliG and FliY proteins. These antibody screening results provide a rationale for further evaluation of these recombinant flagellar proteins as potential vaccines for chickens to improve food safety as well as investigation of host immune response to C. jejuni.

  6. Prevalence and Distribution of Campylobacter jejuni in Small-Scale Broiler Operations.

    PubMed

    Tangkham, Wannee; Janes, Marlene; LeMieux, Frederick

    2016-01-01

    Campylobacter jejuni has been recognized as one of the most prevalent causes of foodborne bacterial illnesses in humans. Previous studies have focused on the transmission routes of C. jejuni from commercial flock farms to the final retail product. The objective of this study was to determine the prevalence of C. jejuni and Campylobacter spp. in eggshells, live birds, feed, drinking water, and the rearing environment in a small-scale broiler operation. Broilers were raised under two different production systems: (i) environmentally controlled housing and (ii) open-air housing with two replications. Each week, samples were collected from eggshells, bird feces, feed, drinking water, enclosures (vertical walls of bird housing), and feed troughs for enumeration and isolation testing. All samples were plated on modified charcoal-cefoperazone-deoxycholate agar to determine the log CFU per gram and percent prevalence of Campylobacter spp. Isolation of C. jejuni was verified with latex agglutination and hippurate hydrolysis tests. The results from this study suggest that vertical transmission of these bacteria from egg surfaces to newly hatched chicks is not a significant risk factor. The results also suggest that the prevalence of C. jejuni at time of harvest (week 6) was significantly higher (P < 0.05) in the open-air housing broilers than in those in the environmentally controlled housing. Elevated levels of cross-contaminants, especially water and feed, may have played a role in this outcome.

  7. [Isolation of Campylobacter jejuni ATCC 29428 from inoculated fried pork meat and roasted chicken].

    PubMed

    Castillo-Martínez, M L; Sánchez-Sánchez, S; Rodríguez-Montaño, R; Quiñones-Ramírez, E I; Lugo de la Fuente, G; Vázquez-Salinas, C

    1993-01-01

    The human gastroenteritis caused by Campylobacter jejuni in some industrialized countries is higher than gastroenteritis produced by Salmonella and Shigella. This has induced the development of techniques to demonstrate the presence of the microorganism in different foods using some culture media combinations. There is not a method to isolate C. jejuni from roasted chicken and fried pork meat, which are popular foods in México. The sensitivity of two culture media combinations was compared: Rama broth (RB)-Rama agar (RA) and Preston broth (PB)-Skirrow agar (SA) to isolate C. jejuni from these foods. The RB-RA combination demonstrated to be the best one to isolate C. jejuni.

  8. Biofilm spatial organization by the emerging pathogen Campylobacter jejuni: comparison between NCTC 11168 and 81-176 strains under microaerobic and oxygen-enriched conditions.

    PubMed

    Turonova, Hana; Briandet, Romain; Rodrigues, Ramila; Hernould, Mathieu; Hayek, Nabil; Stintzi, Alain; Pazlarova, Jarmila; Tresse, Odile

    2015-01-01

    During the last years, Campylobacter has emerged as the leading cause of bacterial foodborne infections in developed countries. Described as an obligate microaerophile, Campylobacter has puzzled scientists by surviving a wide range of environmental oxidative stresses on foods farm to retail, and thereafter intestinal transit and oxidative damage from macrophages to cause human infection. In this study, confocal laser scanning microscopy (CLSM) was used to explore the biofilm development of two well-described Campylobacter jejuni strains (NCTC 11168 and 81-176) prior to or during cultivation under oxygen-enriched conditions. Quantitative and qualitative appraisal indicated that C. jejuni formed finger-like biofilm structures with an open ultrastructure for 81-176 and a multilayer-like structure for NCTC 11168 under microaerobic conditions (MAC). The presence of motile cells within the biofilm confirmed the maturation of the C. jejuni 81-176 biofilm. Acclimation of cells to oxygen-enriched conditions led to significant enhancement of biofilm formation during the early stages of the process. Exposure to these conditions during biofilm cultivation induced an even greater biofilm development for both strains, indicating that oxygen demand for biofilm formation is higher than for planktonic growth counterparts. Overexpression of cosR in the poorer biofilm-forming strain, NCTC 11168, enhanced biofilm development dramatically by promoting an open ultrastructure similar to that observed for 81-176. Consequently, the regulator CosR is likely to be a key protein in the maturation of C. jejuni biofilm, although it is not linked to oxygen stimulation. These unexpected data advocate challenging studies by reconsidering the paradigm of fastidious requirements for C. jejuni growth when various subpopulations (from quiescent to motile cells) coexist in biofilms. These findings constitute a clear example of a survival strategy used by this emerging human pathogen.

  9. Waterborne Campylobacter jejuni epidemic in a Finnish hospital for rheumatic diseases.

    PubMed

    Rautelin, H; Koota, K; von Essen, R; Jahkola, M; Siitonen, A; Kosunen, T U

    1990-01-01

    A waterborne Campylobacter jejuni outbreak in the Rheumatism Foundation Hospital in Heinola, Finland, in November-December 1986 is described. 32 patients and 62 members of the staff developed gastrointestinal symptoms. C. jejuni heat-stable serotype 45 was isolated from the faeces of 32 enteritis patients and from none of the controls. No other enteropathogens were found. Positive serological responses to C. jejuni acid extract antigen were detected by enzyme immunoassay in 34% of the symptomatic hospital patients, in 40% of the symptomatic staff members, and in 10% of the controls. The clinical course of the illness was mostly mild and self-limited. No striking progress in the arthritis symptoms of the patients was found after the outbreak. The hospital has its own water supply. C. jejuni of the same serotype as the epidemic strain was isolated from the water of the pipeline system. After a careful examination some aged components of the waterworks were found to be responsible for leaks that resulted in the contamination of the water.

  10. Trans-cinnamaldehyde, carvacrol, and eugenol reduce Campylobacter jejuni colonization factors and expression of virulence genes in vitro

    USDA-ARS?s Scientific Manuscript database

    Campylobacter jejuni is a major foodborne pathogen that causes severe gastroenteritis in humans characterized by fever, diarrhea and abdominal cramps. In the human gut, Campylobacter adheres and invades the intestinal epithelium followed by cytolethal distending toxin mediated cell death, and enteri...

  11. Characterization of the porins of Campylobacter jejuni and Campylobacter coli and implications for antibiotic susceptibility.

    PubMed Central

    Page, W J; Huyer, G; Huyer, M; Worobec, E A

    1989-01-01

    The major outer membrane protein was extracted from Campylobacter coli by Triton X-100/EDTA fractionation of cell envelopes. This heat-modifiable protein was shown to have pore-forming activity in black lipid bilayers. The C. coli porin formed a relatively small cation-selective pore with a mean single-channel conductance of 0.53 +/- 0.16 nS in 1.0 M KCl. There was no evidence of oligomer formation, which suggested that each protein monomer formed a pore. Pore-forming activity of the C. coli porin and similarly prepared Campylobacter jejuni porin was also measured in liposome-swelling assays. These results confirmed the cation selectivity of both pores. The C. coli porin formed a small pore, which hindered the penetration of solutes with a molecular weight of 262, and a larger pore, which hindered the penetration of solutes with a molecular weight of 340, in a protein-concentration-dependent manner. C. jejuni formed one size of pore that was slightly larger than the C. coli pore and just permitted the passage of solutes, with a molecular weight of 340. A review of the literature concerning in vitro screening of antimicrobial agents tended to confirm the low permeability of the C. jejuni outer membrane to hydrophilic antimicrobial agents except when the molecules had molecular weights of less than 360. The porins of C. jejuni and C. coli may contribute to intrinsic resistance to antimicrobial agents, whereas alternative (nonporin) routes of antimicrobial agent uptake may be more important determinants of susceptibility to antimicrobial agents. Images PMID:2543277

  12. Characterization of the porins of Campylobacter jejuni and Campylobacter coli and implications for antibiotic susceptibility.

    PubMed

    Page, W J; Huyer, G; Huyer, M; Worobec, E A

    1989-03-01

    The major outer membrane protein was extracted from Campylobacter coli by Triton X-100/EDTA fractionation of cell envelopes. This heat-modifiable protein was shown to have pore-forming activity in black lipid bilayers. The C. coli porin formed a relatively small cation-selective pore with a mean single-channel conductance of 0.53 +/- 0.16 nS in 1.0 M KCl. There was no evidence of oligomer formation, which suggested that each protein monomer formed a pore. Pore-forming activity of the C. coli porin and similarly prepared Campylobacter jejuni porin was also measured in liposome-swelling assays. These results confirmed the cation selectivity of both pores. The C. coli porin formed a small pore, which hindered the penetration of solutes with a molecular weight of 262, and a larger pore, which hindered the penetration of solutes with a molecular weight of 340, in a protein-concentration-dependent manner. C. jejuni formed one size of pore that was slightly larger than the C. coli pore and just permitted the passage of solutes, with a molecular weight of 340. A review of the literature concerning in vitro screening of antimicrobial agents tended to confirm the low permeability of the C. jejuni outer membrane to hydrophilic antimicrobial agents except when the molecules had molecular weights of less than 360. The porins of C. jejuni and C. coli may contribute to intrinsic resistance to antimicrobial agents, whereas alternative (nonporin) routes of antimicrobial agent uptake may be more important determinants of susceptibility to antimicrobial agents.

  13. Conceptual design for spacelab pool boiling experiment

    NASA Technical Reports Server (NTRS)

    Lienhard, J. H.; Peck, R. E.

    1978-01-01

    A pool boiling heat transfer experiment to be incorporated with a larger two-phase flow experiment on Spacelab was designed to confirm (or alter) the results of earth-normal gravity experiments which indicate that the hydrodynamic peak and minimum pool boiling heat fluxes vanish at very low gravity. Twelve small sealed test cells containing water, methanol or Freon 113 and cylindrical heaters of various sizes are to be built. Each cell will be subjected to one or more 45 sec tests in which the surface heat flux on the heaters is increased linearly until the surface temperature reaches a limiting value of 500 C. The entire boiling process will be photographed in slow-motion. Boiling curves will be constructed from thermocouple and electric input data, for comparison with the motion picture records. The conduct of the experiment will require no more than a few hours of operator time.

  14. Survival of Campylobacter jejuni in naturally and artificially contaminated laying hen feces.

    PubMed

    Ahmed, M F M; Schulz, J; Hartung, J

    2013-02-01

    Infected laying hens regularly excrete large amounts of Campylobacter jejuni with their feces, which represent a reservoir of infection within the flock and for animals in the region. However, the knowledge about survival times of C. jejuni in these feces is still scarce. Therefore, orienting laboratory experiments were carried out under controlled conditions to estimate the survival times of C. jejuni both in artificially and naturally contaminated laying hen feces. In 6 different laying hen flocks (3 Campylobacter-free and 3 Campylobacter-positive flocks), fresh excreta were randomly collected and pooled in 20-g samples per flock. In the laboratory, each of the 3 pooled samples from the Campylobacter-free barns were homogenized and mixed with 10 mL of a freshly prepared C. jejuni suspension (3 × 10(8) cfu/mL). The other 3 samples were homogenized only. The 6 samples were stored at 20 ± 1°C and 40 to 60% RH in 2 different incubators. Specimens of 2 g were taken from all 6 samples 1 h after storage and daily at the same time during the next 10 consecutive days and investigated on culturable C. jejuni. The survival times of culturable C. jejuni ranged from 72 to 96 h in artificially inoculated feces and varied from 120 to 144 h in naturally colonized flocks. The flaA typing by RFLP confirmed that the isolates from the artificially contaminated feces were identical with the added strain. A total of 5 different flaA types were identified from the naturally contaminated feces, and survival of these isolates was dependent on flaA type. The demonstrated survival times indicate that contaminated fresh feces are an important reservoir of C. jejuni, representing a permanent source of infection over at least 6 d after excretion. It shows the considerable potential of fresh feces in transmitting the agent within and between flocks during that period. This 6-d span should be considered when poultry manure is applied to land as organic fertilizer.

  15. Differentiation of Campylobacter jejuni and Campylobacter coli Using Multiplex-PCR and High Resolution Melt Curve Analysis

    PubMed Central

    Banowary, Banya; Dang, Van Tuan; Sarker, Subir; Connolly, Joanne H.; Chenu, Jeremy; Groves, Peter; Ayton, Michelle; Raidal, Shane; Devi, Aruna; Vanniasinkam, Thiru; Ghorashi, Seyed A.

    2015-01-01

    Campylobacter spp. are important causes of bacterial gastroenteritis in humans in developed countries. Among Campylobacter spp. Campylobacter jejuni (C. jejuni) and C. coli are the most common causes of human infection. In this study, a multiplex PCR (mPCR) and high resolution melt (HRM) curve analysis were optimized for simultaneous detection and differentiation of C. jejuni and C. coli isolates. A segment of the hippuricase gene (hipO) of C. jejuni and putative aspartokinase (asp) gene of C. coli were amplified from 26 Campylobacter isolates and amplicons were subjected to HRM curve analysis. The mPCR-HRM was able to differentiate between C. jejuni and C. coli species. All DNA amplicons generated by mPCR were sequenced. Analysis of the nucleotide sequences from each isolate revealed that the HRM curves were correlated with the nucleotide sequences of the amplicons. Minor variation in melting point temperatures of C. coli or C. jejuni isolates was also observed and enabled some intraspecies differentiation between C. coli and/or C. jejuni isolates. The potential of PCR-HRM curve analysis for the detection and speciation of Campylobacter in additional human clinical specimens and chicken swab samples was also confirmed. The sensitivity and specificity of the test were found to be 100% and 92%, respectively. The results indicated that mPCR followed by HRM curve analysis provides a rapid (8 hours) technique for differentiation between C. jejuni and C. coli isolates. PMID:26394042

  16. MLST genotypes of Campylobacter jejuni isolated from broiler products, dairy cattle and human campylobacteriosis cases in Lithuania.

    PubMed

    Ramonaite, Sigita; Tamuleviciene, Egle; Alter, Thomas; Kasnauskyte, Neringa; Malakauskas, Mindaugas

    2017-06-15

    Campylobacter (C.) jejuni is the leading cause of human campylobacteriosis worldwide. We performed a molecular epidemiological study to investigate the genetic relationship among C. jejuni strains isolated from human diarrhoeal patients, broiler products and dairy cattle in Lithuania. The C. jejuni isolates from human clinical cases, dairy cattle and broiler products were genotyped using multilocus sequence typing (MLST). Allele numbers for each housekeeping gene, sequence type (ST), and clonal complex (CC) were assigned by submitting the DNA sequences to the C. jejuni MLST database ( http://pubmlst.org/campylobacter ). Based on the obtained sequence data of the housekeeping genes a phylogenetic analysis of the strains was performed and a minimum spanning tree (MST) was calculated. Among the 262 C. jejuni strains (consisting of 43 strains isolated from dairy cattle, 102 strains isolated from broiler products and 117 clinical human C. jejuni strains), 82 different MLST sequence types and 22 clonal complexes were identified. Clonal complexes CC21 and CC353 predominated among the C. jejuni strains. On ST-level, five sequence types (ST-5, ST-21, ST-50, ST-464 and ST-6410) were dominating and these five STs accounted for 35.9% (n = 94) of our isolates. In addition, 51 (19.5%) C. jejuni strains representing 27 (32.9%) STs were reported for the first time in the PubMLST database ( http://pubmlst.org/campylobacter ). The highest Czekanowski index or proportional similarity index (PSI) was calculated for C. jejuni strains isolated from human campylobacteriosis cases and broiler products (PSI = 0.32) suggesting a strong link between broiler strains and human cases. The PSI of dairy cattle and human samples was lower (PSI = 0.11), suggesting a weaker link between bovine strains and human cases. The calculated Simpson's index of all C. jejuni isolates showed a high genetic diversity (D = 0.96). Our results suggest that broiler products are the most important source of

  17. Serotyping of Campylobacter jejuni from an outbreak of enteritis implicating chicken.

    PubMed

    Rosenfield, J A; Arnold, G J; Davey, G R; Archer, R S; Woods, W H

    1985-09-01

    An outbreak of campylobacter enteritis involving 7 of 17 people over a period of 5 days followed a dinner at a restaurant. A chicken casserole dish was implicated with a food-specific attack rate of 58%. Campylobacter jejuni Penner serotype 18/21/29, resistant to metronidazole, was isolated from 3 of 4 symptomatic patients and from three raw fresh chicken samples closely associated with the implicated chicken. Numbers of C. jejuni in the chicken ranged from 5.3 X 10(1) to 7.5 X 10(2) colony forming units per square centimeter of surface area. This is the first outbreak of campylobacter enteritis reported in Australia in which C. jejuni has been isolated from both human and food sources and the isolates serologically confirmed as identical.

  18. The signaling pathway of Campylobacter jejuni-induced Cdc42 activation: Role of fibronectin, integrin beta1, tyrosine kinases and guanine exchange factor Vav2

    PubMed Central

    2011-01-01

    Background Host cell invasion by the foodborne pathogen Campylobacter jejuni is considered as one of the primary reasons of gut tissue damage, however, mechanisms and key factors involved in this process are widely unclear. It was reported that small Rho GTPases, including Cdc42, are activated and play a role during invasion, but the involved signaling cascades remained unknown. Here we utilised knockout cell lines derived from fibronectin-/-, integrin-beta1-/-, focal adhesion kinase (FAK)-/- and Src/Yes/Fyn-/- deficient mice, and wild-type control cells, to investigate C. jejuni-induced mechanisms leading to Cdc42 activation and bacterial uptake. Results Using high-resolution scanning electron microscopy, GTPase pulldowns, G-Lisa and gentamicin protection assays we found that each studied host factor is necessary for induction of Cdc42-GTP and efficient invasion. Interestingly, filopodia formation and associated membrane dynamics linked to invasion were only seen during infection of wild-type but not in knockout cells. Infection of cells stably expressing integrin-beta1 variants with well-known defects in fibronectin fibril formation or FAK signaling also exhibited severe deficiencies in Cdc42 activation and bacterial invasion. We further demonstrated that infection of wild-type cells induces increasing amounts of phosphorylated FAK and growth factor receptors (EGFR and PDGFR) during the course of infection, correlating with accumulating Cdc42-GTP levels and C. jejuni invasion over time. In studies using pharmacological inhibitors, silencing RNA (siRNA) and dominant-negative expression constructs, EGFR, PDGFR and PI3-kinase appeared to represent other crucial components upstream of Cdc42 and invasion. siRNA and the use of Vav1/2-/- knockout cells further showed that the guanine exchange factor Vav2 is required for Cdc42 activation and maximal bacterial invasion. Overexpression of certain mutant constructs indicated that Vav2 is a linker molecule between Cdc42 and

  19. Isolation of Campylobacter fetus subsp. jejuni from migratory waterfowl.

    PubMed Central

    Luechtefeld, N A; Blaser, M J; Reller, L B; Wang, W L

    1980-01-01

    Since the sources from which humans acquire Campylobacter enteritis are only partially known, we studied the frequency of carriage of Campylobacter fetus subsp. jejuni in migratory waterfowl. Cecal contents of various species of wild ducks were cultured on selective media that contained antibiotics to inhibit normal flora. Thirty-five percent of the 445 ducks cultured harbored C. fetus subsp. jejuni. Migratory waterfowl are yet another reservoir for this enteric pathogen and may be of public health importance for humans in the contamination of water or when used as food. PMID:7217334

  20. Survival of Campylobacter jejuni during Stationary Phase: Evidence for the Absence of a Phenotypic Stationary-Phase Response

    PubMed Central

    Kelly, Alison F.; Park, Simon F.; Bovill, Richard; Mackey, Bernard M.

    2001-01-01

    When Campylobacter jejuni NCTC 11351 was grown microaerobically in rich medium at 39°C, entry into stationary phase was followed by a rapid decline in viable numbers to leave a residual population of 1% of the maximum number or less. Loss of viability was preceded by sublethal injury, which was seen as a loss of the ability to grow on media containing 0.1% sodium deoxycholate or 1% sodium chloride. Resistance of cells to mild heat stress (50°C) or aeration was greatest in exponential phase and declined during early stationary phase. These results show that C. jejuni does not mount the normal phenotypic stationary-phase response which results in enhanced stress resistance. This conclusion is consistent with the absence of rpoS homologues in the recently reported genome sequence of this species and their probable absence from strain NCTC 11351. During prolonged incubation of C. jejuni NCTC 11351 in stationary phase, an unusual pattern of decreasing and increasing heat resistance was observed that coincided with fluctuations in the viable count. During stationary phase of Campylobacter coli UA585, nonmotile variants and those with impaired ability to form coccoid cells were isolated at high frequency. Taken together, these observations suggest that stationary-phase cultures of campylobacters are dynamic populations and that this may be a strategy to promote survival in at least some strains. Investigation of two spontaneously arising variants (NM3 and SC4) of C. coli UA585 showed that a reduced ability to form coccoid cells did not affect survival under nongrowth conditions. PMID:11319108

  1. Campylobacter jejuni chromosomal sequences that hybridize to Vibrio cholerae and Escherichia coli LT enterotoxin genes.

    PubMed

    Calva, E; Torres, J; Vázquez, M; Angeles, V; de la Vega, H; Ruíz-Palacios, G M

    1989-02-20

    Campylobacter jejuni is one of the main etiologic agents of gastrointestinal illness in developing and developed areas throughout the world. Isolation of enterotoxin-producing C. jejuni has been associated with clinical symptoms of a watery-secretory type of diarrhea. Although physiological and immunological relatedness has been demonstrated between the C. jejuni enterotoxin (CJT), the Vibrio cholerae enterotoxin (CT), and the heat-labile cholera-like Escherichia coli enterotoxin (LT), nucleotide sequence similarity between C. jejuni DNA and either the toxA, toxB, eltA or eltB genes remained to be shown. We found that binding to ganglioside GM1 prevented recognition of CJT by monoclonal antibodies directed to either CT or LT. This indicates antigenic similarity between the three enterotoxins in the ganglioside GM1-binding site. Therefore we searched for corresponding similarities at the DNA level and found, by oligodeoxynucleotide hybridization, C. jejuni chromosomal nucleotide sequences similar to the coding region for a postulated ganglioside GM1-binding site on toxB and eltB.

  2. Distinct Campylobacter jejuni capsular types are related to Guillain-Barré syndrome in The Netherlands and Bangladesh

    USDA-ARS?s Scientific Manuscript database

    An infection with the intestinal pathogen Campylobacter jejuni leads to Guillain-Barré syndrome (GBS) in around one in thousand cases. It is established that sialylated lipooligosaccharides (LOS) of C. jejuni are a crucial virulence factor in GBS development. Frequent detection of C. jejuni with sia...

  3. Transfer of Campylobacter jejuni from raw to cooked chicken via wood and plastic cutting boards.

    PubMed

    Tang, J Y H; Nishibuchi, M; Nakaguchi, Y; Ghazali, F M; Saleha, A A; Son, R

    2011-06-01

    We quantified Campylobacter jejuni transferred from naturally contaminated raw chicken fillets and skins to similar cooked chicken parts via standard rubberwood (RW) and polyethylene cutting boards (PE). RW and PE cutting boards (2.5 × 2.5 cm(2)) were constructed. RW surfaces were smooth and even, whereas PE was uneven. Scoring with scalpel blades produced crevices on RW and flaked patches on the PE boards. Raw chicken breast fillets or skin pieces (10 g) naturally contaminated with Camp. jejuni were used to contaminate the cutting boards (6.25 cm(2)). These were then briefly covered with pieces of cooked chicken. Campylobacter jejuni on raw chicken, the boards, and cooked chicken pieces were counted using a combined most-probable-number (MPN)-PCR method. The type of cutting board (RW, PE; unscored and scored) and temperature of cooked chicken fillets and skins were examined. Unscored PE and RW boards were not significantly different in regards to the mean transfer of Camp. jejuni from raw samples to the boards. The mean transfer of Camp. jejuni from scored RW was significantly higher than from scored PE. When the chicken fillets were held at room temperature, the mean transfer of Camp. jejuni from scored RW and PE was found to be 44.9 and 40.3%, respectively.   RW and PE cutting boards are potential vehicles for Camp. jejuni to contaminate cooked chicken. Although cooked chicken maintained at high temperatures reduced cross-contamination via contaminated boards, a risk was still present. Contamination of cooked chicken by Camp. jejuni from raw chicken via a cutting board is influenced by features of the board (material, changes caused by scoring) and chicken (types of chicken parts and temperature of the cooked chicken). © 2011 The Authors. Letters in Applied Microbiology © 2011 The Society for Applied Microbiology.

  4. Sulfonamide Resistance in Clinical Isolates of Campylobacter jejuni: Mutational Changes in the Chromosomal Dihydropteroate Synthase

    PubMed Central

    Gibreel, Amera; Sköld, Ola

    1999-01-01

    The characterization of the genetic basis of sulfonamide resistance in Campylobacter jejuni was attempted. The resistance determinant from a sulfonamide-resistant strain of C. jejuni was cloned and was found to show 42% identity with the folP gene (which codes for dihydropteroate synthase, the target of sulfonamides) of the related bacterium Helicobacter pylori. The sequences of the areas surrounding the folP gene in C. jejuni showed similarity to those of the areas surrounding the corresponding gene in H. pylori. The folP gene of C. jejuni, which mediates the resistance, was observed to show particular features when it was compared to other known folP genes. One of these features is the presence of two pairs of direct repeats (15 and 27 bp) within the coding sequence of the gene. Comparison of the C. jejuni folP genes that mediate susceptibility and resistance revealed the occurrence of mutations that changed four amino acid residues. Resistance of C. jejuni to sulfonamides could be associated with one or several of these four mutational substitutions, which all occurred in the five different resistant isolates studied. The codon for one of these changed amino acids was found to be located in the second direct repeat within the coding sequence of the gene. The change made the repeat perfect. The transformation of both the resistance and the susceptibility variants of the gene into an Escherichia coli folP knockout mutant was found to complement the dihydropteroate synthase deficiency, confirming that the characterized sulfonamide resistance determinant codes for the C. jejuni dihydropteroate synthase enzyme. Kinetic measurements established different affinities of sulfonamide for the dihydropteroate synthase enzyme isolated from the resistant and susceptible strains. In conclusion, sulfonamide resistance in C. jejuni was shown to be associated with mutational changes in the chromosomally located gene for dihydropteroate synthase, the target of sulfonamides. PMID

  5. Increased intracellular calcium level and impaired nutrient absorption are important pathogenicity traits in the chicken intestinal epithelium during Campylobacter jejuni colonization.

    PubMed

    Awad, Wageha A; Smorodchenko, Alina; Hess, Claudia; Aschenbach, Jörg R; Molnár, Andor; Dublecz, Károly; Khayal, Basel; Pohl, Elena E; Hess, Michael

    2015-08-01

    Although a high number of chickens carry Campylobacter jejuni, the mechanistic action of colonization in the intestine is still poorly understood. The current study was therefore designed to investigate the effects of C. jejuni on glucose uptake, amino acids availability in digesta, and intracellular calcium [Ca(2+)]i signaling in the intestines of broiler chickens. For this, we compared: control birds (n = 60) and C. jejuni-infected birds (n = 60; infected orally with 1 × 10(8) CFU of C. jejuni NCTC 12744 at 14 days of age). Our results showed that glucose uptake was reduced due to C. jejuni infection in isolated jejunal, but not in cecal mucosa at 14 days postinfection (dpi). The decrease in intestinal glucose absorption coincided with a decrease in body weight gain during the 2-week post-infectious period. A reduction in the amount of the amino acids (serine, proline, valine, leucine, phenylalanine, arginine, histidine, and lysine) in ileal digesta of the infected birds at 2 and/or 7 dpi was found, indicating that Campylobacter utilizes amino acids as a carbon source for their multiplication. Applying the cell-permeable Ca(2+) indicator Fluo-4 and two-photon microscopy, we revealed that [Ca(2+)]i was increased in the jejunal and cecal mucosa of infected birds. The muscarinic agonist carbachol induced an increase in [Ca(2+)]i in jejunum and cecum mucosa of control chickens, a response absent in the mucosa of infected chickens, demonstrating that the modulation of [Ca(2+)]i by Campylobacter might be involved in facilitating the necessary cytoskeletal rearrangements that occur during the bacterial invasion of epithelial cells. In conclusion, this study demonstrates the multifaceted interactions of C. jejuni with the gastrointestinal mucosa of broiler chickens. For the first time, it could be shown that a Campylobacter infection could interfere with intracellular Ca(2+) signaling and nutrient absorption in the small intestine with consequences on

  6. Antiadhesion activity of juniper (Juniperus communis L.) preparations against Campylobacter jejuni evaluated with PCR-based methods.

    PubMed

    Klančnik, Anja; Zorko, Špela; Toplak, Nataša; Kovač, Minka; Bucar, Franz; Jeršek, Barbara; Smole Možina, Sonja

    2018-03-01

    The food-borne pathogen Campylobacter jejuni can cause bacterial gastrointestinal infections. Biofilm formation amplifies the risk of human infection by improving survival and persistence of C. jejuni in food processing environments and its transmission through the food chain. We aimed to control C. jejuni using an alternative strategy of low doses of Juniperus communis fruit preparations to target bacterial adhesion properties in the first step of biofilm formation. First, we defined the anti-Campylobacter activity of a juniper fruit crude extract and its fractionated biflavonoids, flavone glycosides, and purified amentoflavone, of juniper fruit essential oil and of juniper fruit postdistillation waste material extract. For accurate quantification of adherent C. jejuni, we optimised digital Polymerase Chain Reaction (PCR) and quantitative real-time PCR for construction of standard curves and quantification. We show for the first time that juniper fruit formulations can effectively inhibit adhesion of C. jejuni to polystyrene. Furthermore, ≥94% of the antiadhesion activity of juniper fruit crude extract and juniper fruit essential oil remained under food-related conditions: modified culture medium with glucose, or a stainless steel surface, or mixed co-cultures of C. jejuni and Listeria monocytogenes. This study indicates that addition of juniper fruit formulations can control growth and adhesion of C. jejuni and thus limit food chain transmission of campylobacters. Copyright © 2017 John Wiley & Sons, Ltd.

  7. Serum antibodies against gangliosides and Campylobacter jejuni lipopolysaccharides in Miller Fisher syndrome.

    PubMed Central

    Neisser, A; Bernheimer, H; Berger, T; Moran, A P; Schwerer, B

    1997-01-01

    Seven patients with Miller Fisher syndrome (MFS), six in the acute phase and one in the recovery phase, were investigated for serum antibodies against gangliosides and purified lipopolysaccharides (LPS) from different strains of Campylobacter jejuni, including the MFS-associated serotypes O:2 and O:23. Immunoglobulin G antibodies against gangliosides GT1a and GQ1b were found in five of six patients in the acute phase of disease. Three of these patients also displayed antibodies to ganglioside GD2, a finding not previously reported for MFS. All anti-GT1a- and anti-GQ1b-seropositive patients showed antibody binding to C. jejuni LPS, predominantly to O:2 and O:23 LPS. Antibody cross-reactivity between gangliosides GT1a and GQ1b and O:2 and O:23 LPS was demonstrated by adsorption studies. This cross-reactivity between gangliosides and C.jejuni LPS, which is obviously due to oligosaccharide homologies, may be an important pathogenetic factor in the development of MFS after C. jejuni infection. PMID:9317004

  8. Rapid Evolution and the Importance of Recombination to the Gastroenteric Pathogen Campylobacter jejuni

    PubMed Central

    Gabriel, Edith; Leatherbarrow, Andrew J.H.; Cheesbrough, John; Gee, Steven; Bolton, Eric; Fox, Andrew; Hart, C. Anthony; Diggle, Peter J.; Fearnhead, Paul

    2009-01-01

    Responsible for the majority of bacterial gastroenteritis in the developed world, Campylobacter jejuni is a pervasive pathogen of humans and animals, but its evolution is obscure. In this paper, we exploit contemporary genetic diversity and empirical evidence to piece together the evolutionary history of C. jejuni and quantify its evolutionary potential. Our combined population genetics–phylogenetics approach reveals a surprising picture. Campylobacter jejuni is a rapidly evolving species, subject to intense purifying selection that purges 60% of novel variation, but possessing a massive evolutionary potential. The low mutation rate is offset by a large effective population size so that a mutation at any site can occur somewhere in the population within the space of a week. Recombination has a fundamental role, generating diversity at twice the rate of de novo mutation, and facilitating gene flow between C. jejuni and its sister species Campylobacter coli. We attempt to calibrate the rate of molecular evolution in C. jejuni based solely on within-species variation. The rates we obtain are up to 1,000 times faster than conventional estimates, placing the C. jejuni–C. coli split at the time of the Neolithic revolution. We weigh the plausibility of such recent bacterial evolution against alternative explanations and discuss the evidence required to settle the issue. PMID:19008526

  9. Effect of organic acids and marination ingredients on the survival of Campylobacter jejuni on meat.

    PubMed

    Birk, Tina; Grønlund, Anne Christine; Christensen, Bjarke Bak; Knøchel, Susanne; Lohse, Kristin; Rosenquist, Hanne

    2010-02-01

    The aim of this study was to determine whether marination of chicken meat in different food ingredients can be used to reduce populations of Campylobacter jejuni. C. jejuni strains were exposed to different organic acids (tartaric, acetic, lactic, malic, and citric acids) and food marinating ingredients at 4 degrees C in broth and on chicken meat. The organic acids (0.5%) reduced populations of C. jejuni in broth (chicken juice and brain heart infusion broth) by 4 to 6 log units (after 24 h); tartaric acid was the most efficient treatment. Large strain variation was observed among 14 C. jejuni isolates inoculated in brain heart infusion broth containing 0.3% tartaric acid. On chicken meat medallions, reductions of C. jejuni were 0.5 to 2 log units when tartaric acid solutions (2, 4, 6, and 10%) were spread onto the meat. Analysis of acidic food ingredient (e.g., vinegar, lemon juice, pomegranate syrup, and soya sauce) revealed that such ingredients reduced counts of C. jejuni by at least 0.8 log units on meat medallions. Three low pH marinades (pH < 3) based on pomegranate syrup, lemon juice, and white wine vinegar were prepared. When applied to whole filets, these marinades resulted in a reduction of approximately 1.2 log units after 3 days of storage. Taste evaluations of chicken meat that had been marinated and then fried were graded positively for flavor and texture. Thus, success was achieved in creating a marinade with an acceptable taste that reduced the counts of C. jejuni.

  10. Morphological control in polymer solar cells using low-boiling-point solvent additives

    NASA Astrophysics Data System (ADS)

    Mahadevapuram, Rakesh C.

    In the global search for clean, renewable energy sources, organic photovoltaics (OPVs) have recently been given much attention. Popular modern-day OPVs are made from solution-processible, carbon-based polymers (e.g. the model poly(3-hexylthiophene) that are intimately blended with fullerene derivatives (e.g. [6,6]-phenyl-C71-butyric acid methyl ester) to form what is known as the dispersed bulk-heterojunction (BHJ). This BHJ architecture has produced some of the most efficient OPVs to date, with reports closing in on 10% power conversion efficiency. To push efficiencies further into double digits, many groups have identified the BHJ nanomorphology---that is, the phase separations and grain sizes within the polymer: fullerene composite---as a key aspect in need of control and improvement. As a result, many methods, including thermal annealing, slow-drying (solvent) annealing, vapor annealing, and solvent additives, have been developed and studied to promote BHJ self-organization. Processing organic photovoltaic (OPV) blend solutions with high-boiling-point solvent additives has recently been used for morphological control in BHJ OPV cells. Here we show that even low-boiling-point solvents can be effective additives. When P3HT:PCBM OPV cells were processed with a low-boiling-point solvent tetrahydrafuran as an additive in parent solvent o-dichlorobenzene, charge extraction increased leading to fill factors as high as 69.5%, without low work-function cathodes, electrode buffer layers or thermal treatment. This was attributed to PCBM demixing from P3HT domains and better vertical phase separation, as indicated by photoluminescence lifetimes, hole mobilities, and shunt leakage currents. Dependence on solvent parameters and applicability beyond P3HT system was also investigated.

  11. In Vivo Phase Variation and Serologic Response to Lipooligosaccharide of Campylobacter jejuni in Experimental Human Infection

    DTIC Science & Technology

    2004-02-01

    pregnancy, HLA B27 positivity, human im- munodeficiency virus seropositivity, or any disease or condition as determined by a study physician that would place...leukocyte antigen ( HLA ) types and the development of C. jejuni-associated GBS have been inconclusive (21). Thus, the requirements for an individual to be...susceptible to GBS development after C. jejuni infection remain to be identified since HLA type has not been consistently correlated with C. jejuni

  12. Methods to Assess the Direct Interaction of C. jejuni with Mucins.

    PubMed

    Clyne, Marguerite; Duggan, Gina; Naughton, Julie; Bourke, Billy

    2017-01-01

    Studies of the interaction of bacteria with mucus-secreting cells can be complemented at a more mechanistic level by exploring the interaction of bacteria with purified mucins. Here we describe a far Western blotting approach to show how C. jejuni proteins separated by SDS PAGE and transferred to a membrane or slot blotted directly onto a membrane can be probed using biotinylated mucin. In addition we describe the use of novel mucin microarrays to assess bacterial interactions with mucins in a high-throughput manner.

  13. Coadministration of the Campylobacter jejuni N-Glycan-Based Vaccine with Probiotics Improves Vaccine Performance in Broiler Chickens

    PubMed Central

    Perez-Muñoz, M. E.; Gouveia, G. J.; Wanford, J. J.; Lango-Scholey, L.; Panagos, C. G.; Srithayakumar, V.; Plastow, G. S.; Coros, C.; Bayliss, C. D.; Edison, A. S.; Walter, J.

    2017-01-01

    ABSTRACT Source attribution studies report that the consumption of contaminated poultry is the primary source for acquiring human campylobacteriosis. Oral administration of an engineered Escherichia coli strain expressing the Campylobacter jejuni N-glycan reduces bacterial colonization in specific-pathogen-free leghorn chickens, but only a fraction of birds respond to vaccination. Optimization of the vaccine for commercial broiler chickens has great potential to prevent the entry of the pathogen into the food chain. Here, we tested the same vaccination approach in broiler chickens and observed similar efficacies in pathogen load reduction, stimulation of the host IgY response, the lack of C. jejuni resistance development, uniformity in microbial gut composition, and the bimodal response to treatment. Gut microbiota analysis of leghorn and broiler vaccine responders identified one member of Clostridiales cluster XIVa, Anaerosporobacter mobilis, that was significantly more abundant in responder birds. In broiler chickens, coadministration of the live vaccine with A. mobilis or Lactobacillus reuteri, a commonly used probiotic, resulted in increased vaccine efficacy, antibody responses, and weight gain. To investigate whether the responder-nonresponder effect was due to the selection of a C. jejuni “supercolonizer mutant” with altered phase-variable genes, we analyzed all poly(G)-containing loci of the input strain compared to nonresponder colony isolates and found no evidence of phase state selection. However, untargeted nuclear magnetic resonance (NMR)-based metabolomics identified a potential biomarker negatively correlated with C. jejuni colonization levels that is possibly linked to increased microbial diversity in this subgroup. The comprehensive methods used to examine the bimodality of the vaccine response provide several opportunities to improve the C. jejuni vaccine and the efficacy of any vaccination strategy. IMPORTANCE Campylobacter jejuni is a common

  14. The Campylobacter jejuni Oxidative Stress Regulator RrpB Is Associated with a Genomic Hypervariable Region and Altered Oxidative Stress Resistance.

    PubMed

    Gundogdu, Ozan; da Silva, Daiani T; Mohammad, Banaz; Elmi, Abdi; Wren, Brendan W; van Vliet, Arnoud H M; Dorrell, Nick

    2016-01-01

    Campylobacter jejuni is the leading cause of bacterial foodborne diarrhoeal disease worldwide. Despite the microaerophilic nature of the bacterium, C. jejuni can survive the atmospheric oxygen conditions in the environment. Bacteria that can survive either within a host or in the environment like C. jejuni require variable responses to survive the stresses associated with exposure to different levels of reactive oxygen species. The MarR-type transcriptional regulators RrpA and RrpB have recently been shown to play a role in controlling both the C. jejuni oxidative and aerobic stress responses. Analysis of 3,746 C. jejuni and 486 C. coli genome sequences showed that whilst rrpA is present in over 99% of C. jejuni strains, the presence of rrpB is restricted and appears to correlate with specific MLST clonal complexes (predominantly ST-21 and ST-61). C. coli strains in contrast lack both rrpA and rrpB . In C. jejuni rrpB + strains, the rrpB gene is located within a variable genomic region containing the IF subtype of the type I Restriction-Modification ( hsd ) system, whilst this variable genomic region in C. jejuni rrpB - strains contains the IAB subtype hsd system and not the rrpB gene. C. jejuni rrpB - strains exhibit greater resistance to peroxide and aerobic stress than C. jejuni rrpB + strains. Inactivation of rrpA resulted in increased sensitivity to peroxide stress in rrpB + strains, but not in rrpB - strains. Mutation of rrpA resulted in reduced killing of Galleria mellonella larvae and enhanced biofilm formation independent of rrpB status. The oxidative and aerobic stress responses of rrpB - and rrpB + strains suggest adaptation of C. jejuni within different hosts and niches that can be linked to specific MLST clonal complexes.

  15. Methods for Isolation, Purification, and Propagation of Bacteriophages of Campylobacter jejuni.

    PubMed

    Gencay, Yilmaz Emre; Birk, Tina; Sørensen, Martine Camilla Holst; Brøndsted, Lone

    2017-01-01

    Here, we describe the methods for isolation, purification, and propagation of Campylobacter jejuni bacteriophages from samples expected to contain high number of phages such as chicken feces. The overall steps are (1) liberation of phages from the sample material; (2) observation of plaque-forming units on C. jejuni lawns using a spot assay; (3) isolation of single plaques; (4) consecutive purification procedures; and (5) propagation of purified phages from a plate lysate to prepare master stocks.

  16. [Campylobacter jejuni O:19 serotype in Argentine poultry meat supply chain].

    PubMed

    Rossler, Eugenia; Fuhr, Estefanía M; Lorenzón, Guillermina; Romero-Scharpen, Analía; Berisvil, Ayelén P; Blajman, Jesica E; Astesana, Diego M; Zimmermann, Jorge A; Fusari, Marcia L; Signorini, Marcelo L; Soto, Lorena P; Frizzo, Laureano S; Zbrun, María V

    Thermotolerant species of Campylobacter have been focus of attention in the last years because they are the major agent causing zoonotic foodborne diseases. In addition, Campylobacter jejuni O:19 serotype was associated with Guillain Barré syndrome. The aim of this study was to determine the proportion of C. jejuni O:19 serotype isolated at different stages of three poultry meat supply chain in Santa Fe, Argentina. The analysis showed that 18% of isolated C. jejuni belong to serotype O:19. It was also determined that the presence of these strains is given in almost all production stages. These results reflect a significant risk to public health of consumers. Epidemiological studies of Campylobacter should be considered to establish a risk manager policy. Copyright © 2017 Asociación Argentina de Microbiología. Publicado por Elsevier España, S.L.U. All rights reserved.

  17. Detection of Campylobacter jejuni in Lizard Faeces from Central Australia Using Quantitative PCR

    PubMed Central

    Whiley, Harriet; McLean, Ryan; Ross, Kirstin

    2016-01-01

    Worldwide, Campylobacter is a significant cause of gastrointestinal illness. It is predominately considered a foodborne pathogen, with human exposure via non-food transmission routes generally overlooked. Current literature has been exploring environmental reservoirs of campylobacteriosis including potential wildlife reservoirs. Given the close proximity between lizards and human habitats in Central Australia, this study examined the presence of Campylobacter jejuni from lizard faeces collected from this region. Of the 51 samples collected, 17 (33%) (this included 14/46 (30%) wild and 3/5 (60%) captive lizard samples) were positive for C. jejuni using quantitative PCR (qPCR). This was the first study to investigate the presence of C. jejuni in Australian lizards. This has public health implications regarding the risk of campylobacteriosis from handling of pet reptiles and through cross-contamination or contact with wild lizard faeces. Additionally this has implication for horizontal transmission via lizards of C. jejuni to food production farms. Further research is needed on this environmental reservoir and potential transmission routes to reduce the risk to public health. PMID:28025556

  18. Campylobacter jejuni transcriptome changes during loss of culturability in water

    PubMed Central

    Bronowski, Christina; Mustafa, Kasem; Goodhead, Ian; James, Chloe E.; Nelson, Charlotte; Lucaci, Anita; Wigley, Paul; Humphrey, Tom J.; Williams, Nicola J.; Winstanley, Craig

    2017-01-01

    Background Water serves as a potential reservoir for Campylobacter, the leading cause of bacterial gastroenteritis in humans. However, little is understood about the mechanisms underlying variations in survival characteristics between different strains of C. jejuni in natural environments, including water. Results We identified three Campylobacter jejuni strains that exhibited variability in their ability to retain culturability after suspension in tap water at two different temperatures (4°C and 25°C). Of the three, strains C. jejuni M1 exhibited the most rapid loss of culturability whilst retaining viability. Using RNAseq transcriptomics, we characterised C. jejuni M1 gene expression in response to suspension in water by analyzing bacterial suspensions recovered immediately after introduction into water (Time 0), and from two sampling time/temperature combinations where considerable loss of culturability was evident, namely (i) after 24 h at 25°C, and (ii) after 72 h at 4°C. Transcript data were compared with a culture-grown control. Some gene expression characteristics were shared amongst the three populations recovered from water, with more genes being up-regulated than down. Many of the up-regulated genes were identified in the Time 0 sample, whereas the majority of down-regulated genes occurred in the 25°C (24 h) sample. Conclusions Variations in expression were found amongst genes associated with oxygen tolerance, starvation and osmotic stress. However, we also found upregulation of flagellar assembly genes, accompanied by down-regulation of genes involved in chemotaxis. Our data also suggested a switch from secretion via the sec system to via the tat system, and that the quorum sensing gene luxS may be implicated in the survival of strain M1 in water. Variations in gene expression also occurred in accessory genome regions. Our data suggest that despite the loss of culturability, C. jejuni M1 remains viable and adapts via specific changes in gene

  19. Comparative quantification of Campylobacter jejuni from environmental samples using traditional and molecular biological techniques

    USDA-ARS?s Scientific Manuscript database

    Campylobacter jejuni (C. jejuni) is one of the most common causes of gastroenteritis in the world. Given the potential risks to human, animal and environmental health the development and optimization of methods to quantify this important pathogen in environmental samples is essential. Two of the mos...

  20. Methods for Initial Characterization of Campylobacter jejuni Bacteriophages.

    PubMed

    Sørensen, Martine Camilla Holst; Gencay, Yilmaz Emre; Brøndsted, Lone

    2017-01-01

    Here we describe an initial characterization of Campylobacter jejuni bacteriophages by host range analysis, genome size determination by pulsed-field gel electrophoresis, and receptor-type identification by screening mutants for phage sensitivity.

  1. [The occurrence of campylobacter fetus subsp. jejuni and Salmonella bacteria in some wild birds (author's transl)].

    PubMed

    Rosef, O

    1981-12-01

    An investigation was carried out into the occurrence of Campylobacter fetus subsp. jejuni and Salmonella species in some wild birds. A total of 129 birds was examined, consisting of 71 pigeons, 54 seagulls, three crows and one raven. Campylobacter bacteria were isolated from 32 birds (24.8%), of which three were pigeons, 27 seagulls and two were crows. Of the 27 Campylobacter strains isolated from seagulls, four had the biochemical characteristics of the NARTC biotype described by Skirrow and Benjamin, seven were grouped as Campylobacter coli biotype and 16 as the biotype of Campylobacter jejuni. All the strains isolated from crows and pigeons had the biochemical characteristics of Campylobacter jejuni biotypes. Salmonella bacteria were isolated from the intestinal contents of two of the 54 seagulls (3.7%), and were identified serologically as Salmonella indiana and Salmonella typhimurium. One seagull was found to be a carrier of both Campylobacter fetus subsp. jejuni and Salmonella typhimurium. A correlation could not be demonstrated between the occurrence of Salmonella bacteria and Campylobacter fetus subsp. jejuni.

  2. Boiling incipience and convective boiling of neon and nitrogen

    NASA Technical Reports Server (NTRS)

    Papell, S. S.; Hendricks, R. C.

    1977-01-01

    Forced convection and subcooled boiling heat transfer data for liquid nitrogen and liquid neon were obtained in support of a design study for a 30 tesla cryomagnet cooled by forced convection of liquid neon. This design precludes nucleate boiling in the flow channels as they are too small to handle vapor flow. Consequently, it was necessary to determine boiling incipience under the operating conditions of the magnet system. The cryogen data obtained over a range of system pressures, fluid flow rates, and applied heat fluxes were used to develop correlations for predicting boiling incipience and convective boiling heat transfer coefficients in uniformly heated flow channels. The accuracy of the correlating equations was then evaluated. A technique was also developed to calculate the position of boiling incipience in a uniformly heated flow channel. Comparisons made with the experimental data showed a prediction accuracy of plus or minus 15 percent

  3. Campylobacter jejuni in Musca domestica: An examination of survival and transmission potential in light of the innate immune responses of the house flies.

    PubMed

    Gill, Carson; Bahrndorff, Simon; Lowenberger, Carl

    2017-08-01

    The house fly, Musca domestica, has been implicated as a vector of Campylobacter spp., a major cause of human disease. Little is known whether house flies serve as biological amplifying hosts or mechanical vectors for Campylobacter jejuni. We investigated the period after C. jejuni had been ingested by house flies in which viable C. jejuni colonies could be isolated from whole bodies, the vomitus and the excreta of adult M. domestica and evaluated the activation of innate immune responses of house flies to ingested C. jejuni over time. C. jejuni could be cultured from infected houseflies soon after ingestion but no countable C. jejuni colonies were observed > 24 h postingestion. We detected viable C. jejuni in house fly vomitus and excreta up to 4 h after ingestion, but no viable bacteria were detected ≥ 8 h. Suppression subtractive hybridization identified pathogen-induced gene expression in the intestinal tracts of adult house flies 4-24 h after ingesting C. jejuni. We measured the expression of immune regulatory (thor, JNK, and spheroide) and effector (cecropin, diptericin, attacin, defensing, and lysozyme) genes in C. jejuni-infected and -uninfected house flies using quantitative real time PCR. Some house fly factor, or combination of factors, eliminates C. jejuni within 24 h postingestion. Because C. jejuni is not amplified within the body of the housefly, this insect likely serves as a mechanical vector rather than as a true biological, amplifying vector for C. jejuni, and adds to our understanding of insect-pathogen interactions. © 2016 Institute of Zoology, Chinese Academy of Sciences.

  4. Comparison of Proteomics Profiles of Campylobacter jejuni Strain Bf under Microaerobic and Aerobic Conditions

    PubMed Central

    Rodrigues, Ramila C.; Haddad, Nabila; Chevret, Didier; Cappelier, Jean-Michel; Tresse, Odile

    2016-01-01

    Campylobacter jejuni accounts for one of the leading causes of foodborne bacterial enteritis in humans. Despite being considered an obligate microaerobic microorganism, C. jejuni is regularly exposed to oxidative stress. However, its adaptive strategies to survive the atmospheric oxygen level during transmission to humans remain unclear. Recently, the clinical C. jejuni strain Bf was singled out for its unexpected ability to grow under ambient atmosphere. Here, we aimed to understand better the biological mechanisms underlying its atypical aerotolerance trait using two-dimensional protein electrophoresis, gene expression, and enzymatic activities. Forty-seven proteins were identified with a significantly different abundance between cultivation under microaerobic and aerobic conditions. The over-expressed proteins in aerobiosis belonged mainly to the oxidative stress response, enzymes of the tricarboxylic acid cycle, iron uptake, and regulation, and amino acid uptake when compared to microaerobic conditions. The higher abundance of proteins related to oxidative stress was correlated to dramatically higher transcript levels of the corresponding encoding genes in aerobic conditions compared to microaerobic conditions. In addition, a higher catalase-equivalent activity in strain Bf was observed. Despite the restricted catabolic capacities of C. jejuni, this study reveals that strain Bf is equipped to withstand oxidative stress. This ability could contribute to emergence and persistence of particular strains of C. jejuni throughout food processing or macrophage attack during human infection. PMID:27790195

  5. Administration of Lactobacillus johnsonii FI9785 to chickens affects colonisation by Campylobacter jejuni and the intestinal microbiota.

    PubMed

    Mañes-Lázaro, R; Van Diemen, P M; Pin, C; Mayer, M J; Stevens, M P; Narbad, A

    2017-08-01

    1. Campylobacter jejuni is the most common bacterial cause of human food-borne gastroenteritis in the world. A major source of human infection is the consumption of contaminated meat, particularly poultry. New control measures to reduce or eliminate this pathogen from the animal gastrointestinal tract are urgently required, and the use of probiotics as competitive exclusion agents is a promising biocontrol measure to reduce C. jejuni in the food chain. 2. In this study, we assessed the potential of Lactobacillus johnsonii FI9785, which has shown efficacy against Clostridium perfringens, to combat C. jejuni. The effect of prophylactic administration of L. johnsonii on the ability of C. jejuni to colonise chickens was determined. 3. Two doses of L. johnsonii given a week apart led to a reduction in C. jejuni colonisation in the caecal contents, but this biocontrol seemed reliant upon a high level of initial colonisation by the probiotic. 4. The microbial composition in the chicken gut was significantly altered by the probiotic treatment, as shown by denaturing gradient gel electrophoresis of 16S rRNA gene amplicons. 5. Together these results demonstrate the potential of this probiotic strain to be tested further as a competitive exclusion agent in poultry against C. jejuni.

  6. Hyperendemic Campylobacter jejuni in guinea pigs (Cavia porcellus) raised for food in a semi-rural community of Quito, Ecuador.

    PubMed

    Graham, Jay P; Vasco, Karla; Trueba, Gabriel

    2016-06-01

    Domestic animals and animal products are the source of pathogenic Campylobacter jejuni and C. coli in industrialized countries, yet little is known about the transmission of these bacteria in developing countries. Guinea pigs (Cavia porcellus) are commonly raised for food in the Andean region of South America, however, limited research has characterized this rodent as a reservoir of zoonotic enteric pathogens. In this study, we examined the prevalence of Campylobacter spp. in 203 fecal samples from domestic animals of 59 households in a semi-rural parish of Quito, Ecuador. Of the twelve animal species studied, guinea pigs showed the highest prevalence of C. jejuni (n = 39/40; 97.5%). Multilocus sequence typing (MLST) was used to characterize the genetic relationship of C. jejuni from domestic animals and 21 sequence types (STs) were identified. The majority of STs from guinea pigs appeared to form new clonal complexes that were not related to STs of C. jejuni isolated from other animal species and shared only a few alleles with other C. jejuni previously characterized. The study identifies guinea pigs as a major reservoir of C. jejuni and suggests that some C. jejuni strains are adapted to this animal species. © 2016 Society for Applied Microbiology and John Wiley & Sons Ltd.

  7. Campylobacter jejuni CsrA Regulates Metabolic and Virulence Associated Proteins and Is Necessary for Mouse Colonization.

    PubMed

    Fields, Joshua A; Li, Jiaqi; Gulbronson, Connor J; Hendrixson, David R; Thompson, Stuart A

    2016-01-01

    Campylobacter jejuni infection is a leading bacterial cause of gastroenteritis and a common antecedent leading to Gullian-Barré syndrome. Our previous data suggested that the RNA-binding protein CsrA plays an important role in regulating several important phenotypes including motility, biofilm formation, and oxidative stress resistance. In this study, we compared the proteomes of wild type, csrA mutant, and complemented csrA mutant C. jejuni strains in an effort to elucidate the mechanisms by which CsrA affects virulence phenotypes. The putative CsrA regulon was more pronounced at stationary phase (111 regulated proteins) than at mid-log phase (25 regulated proteins). Proteins displaying altered expression in the csrA mutant included diverse metabolic functions, with roles in amino acid metabolism, TCA cycle, acetate metabolism, and various other cell processes, as well as pathogenesis-associated characteristics such as motility, chemotaxis, oxidative stress resistance, and fibronectin binding. The csrA mutant strain also showed altered autoagglutination kinetics when compared to the wild type. CsrA specifically bound the 5' end of flaA mRNA, and we demonstrated that CsrA is a growth-phase dependent repressor of FlaA expression. Finally, the csrA mutant exhibited reduced ability to colonize in a mouse model when in competition with the wild type, further underscoring the role of CsrA in C. jejuni colonization and pathogenesis.

  8. Campylobacter jejuni CsrA Regulates Metabolic and Virulence Associated Proteins and Is Necessary for Mouse Colonization

    PubMed Central

    Fields, Joshua A.; Li, Jiaqi; Gulbronson, Connor J.; Hendrixson, David R.

    2016-01-01

    Campylobacter jejuni infection is a leading bacterial cause of gastroenteritis and a common antecedent leading to Gullian-Barré syndrome. Our previous data suggested that the RNA-binding protein CsrA plays an important role in regulating several important phenotypes including motility, biofilm formation, and oxidative stress resistance. In this study, we compared the proteomes of wild type, csrA mutant, and complemented csrA mutant C. jejuni strains in an effort to elucidate the mechanisms by which CsrA affects virulence phenotypes. The putative CsrA regulon was more pronounced at stationary phase (111 regulated proteins) than at mid-log phase (25 regulated proteins). Proteins displaying altered expression in the csrA mutant included diverse metabolic functions, with roles in amino acid metabolism, TCA cycle, acetate metabolism, and various other cell processes, as well as pathogenesis-associated characteristics such as motility, chemotaxis, oxidative stress resistance, and fibronectin binding. The csrA mutant strain also showed altered autoagglutination kinetics when compared to the wild type. CsrA specifically bound the 5’ end of flaA mRNA, and we demonstrated that CsrA is a growth-phase dependent repressor of FlaA expression. Finally, the csrA mutant exhibited reduced ability to colonize in a mouse model when in competition with the wild type, further underscoring the role of CsrA in C. jejuni colonization and pathogenesis. PMID:27257952

  9. High-Throughput Sequencing of Campylobacter jejuni Insertion Mutant Libraries Reveals mapA as a Fitness Factor for Chicken Colonization

    PubMed Central

    Johnson, Jeremiah G.; Livny, Jonathan

    2014-01-01

    Campylobacter jejuni is a leading cause of gastrointestinal infections worldwide, due primarily to its ability to asymptomatically colonize the gastrointestinal tracts of agriculturally relevant animals, including chickens. Infection often occurs following consumption of meat that was contaminated by C. jejuni during harvest. Because of this, much interest lies in understanding the mechanisms that allow C. jejuni to colonize the chicken gastrointestinal tract. To address this, we generated a C. jejuni transposon mutant library that is amenable to insertion sequencing and introduced this mutant pool into day-of-hatch chicks. Following deep sequencing of C. jejuni mutants in the cecal outputs, several novel factors required for efficient colonization of the chicken gastrointestinal tract were identified, including the predicted outer membrane protein MapA. A mutant strain lacking mapA was constructed and found to be significantly reduced for chicken colonization in both competitive infections and monoinfections. Further, we found that mapA is required for in vitro competition with wild-type C. jejuni but is dispensable for growth in monoculture. PMID:24633877

  10. High-throughput sequencing of Campylobacter jejuni insertion mutant libraries reveals mapA as a fitness factor for chicken colonization.

    PubMed

    Johnson, Jeremiah G; Livny, Jonathan; Dirita, Victor J

    2014-06-01

    Campylobacter jejuni is a leading cause of gastrointestinal infections worldwide, due primarily to its ability to asymptomatically colonize the gastrointestinal tracts of agriculturally relevant animals, including chickens. Infection often occurs following consumption of meat that was contaminated by C. jejuni during harvest. Because of this, much interest lies in understanding the mechanisms that allow C. jejuni to colonize the chicken gastrointestinal tract. To address this, we generated a C. jejuni transposon mutant library that is amenable to insertion sequencing and introduced this mutant pool into day-of-hatch chicks. Following deep sequencing of C. jejuni mutants in the cecal outputs, several novel factors required for efficient colonization of the chicken gastrointestinal tract were identified, including the predicted outer membrane protein MapA. A mutant strain lacking mapA was constructed and found to be significantly reduced for chicken colonization in both competitive infections and monoinfections. Further, we found that mapA is required for in vitro competition with wild-type C. jejuni but is dispensable for growth in monoculture.

  11. Prevalence and characterization of Campylobacter jejuni isolated from pasture flock poultry.

    PubMed

    Hanning, Irene; Biswas, Debabrata; Herrera, Paul; Roesler, Mary; Ricke, Steven C

    2010-09-01

    The growing interest in organic and natural foods warrants a greater need for information on the food safety of these products. In this study, samples were taken from 2 pasture flock farms (N = 178; feed, water, drag swabs, and insect traps), pasture flock retail carcasses (N = 48) and 1 pasture flock processing facility (N = 16) over a period of 8 mo. A total of 105 Campylobacter isolates were obtained from 53 (30%), 36 (75%), and 16 (100%) samples from the farms, retail carcasses, and processing facility, respectively. Of the 105 isolates collected, 65 were C. jejuni, 31 were C. coli, and 9 were other Campylobacter spp. Using PCR, the C. jejuni isolates were further analyzed for virulence genes involved in colonization and survival (flaA, flaC, cadF, dnaJ, racR, cbrR), invasion (virB11, ciaB, pldA), protection against harsh conditions (sodB, htrA, clpA), toxin production (cdtA, cdtB, cdtC), siderophore transport (ceuE), and ganglioside mimicry (wlaN). In addition, the short variable region of the flaA locus (flaA SVR) was sequenced to determine the genetic diversity of the C. jejuni isolates. The flaA SVR diversity indices increased along the farm to carcass continuum. PCR-based analysis indicated a low prevalence of 5 genes involved in colonization (dnaJ, ciaB, pldA, racR, virB11). The results of this survey indicate that the prevalence of Campylobacter on organic retail carcasses is similar to prevalence reports of Campylobacter on conventional retail carcasses. However, the genetic diversity of the flaA SVR genotypes increased along the farm to carcass continuum that contrasted with conventional poultry studies. Campylobacter jejuni is a leading cause of foodborne illness with poultry and poultry products being leading sources of infection. Free-range and pasture flock chickens are becoming more popular; however, there is an inherent biosecurity risk that can increase the prevalence of foodborne pathogens in these flocks. This study aimed to determine sources

  12. Presence and characterization of Campylobacter jejuni in organically raised chickens in Quebec

    PubMed Central

    Thibodeau, Alexandre; Fravalo, Philippe; Laurent-Lewandowski, Sylvette; Guévremont, Evelyne; Quessy, Sylvain; Letellier, Ann

    2011-01-01

    The objective of this study was to estimate the presence of the important foodborne pathogen Campylobacter jejuni in organically raised chickens in the province of Quebec. The recovered isolates were further characterized for their antimicrobial resistance profile, autoagglutination property and chemotaxis. Antimicrobial resistance was evaluated using agar dilution for: tetracycline, erythromycin, chloramphenicol, ciprofloxacin, gentamicin, nalidixic acid, clindamycin, ampicillin, azithromycin, bacitracin, and ceftiofur. Autoagglutination was measured by monitoring optical density changes in a bacterial suspension after 3 h of incubation at room temperature. Chemotaxis was evaluated after a contact time of 3 h between isolates and mucin, using a quantitative protocol. A total of 10 lots of chickens was sampled in August and September 2009; half of them were positive for the presence of C. jejuni. Antimicrobial resistance was found only for tetracycline (44%), erythromycin (6%), azithromycin (6%) and clindamycin (2%). Variation was observed in the minimum inhibitory concentrations (MICs) for ceftiofur and bacitracin, for which C. jejuni possess intrinsic resistance. Autoagglutination and chemotaxis varied among isolates and lot-level differences in these were observed. Autoagglutination and chemotaxis levels appeared as independent isolate properties. Further monitoring and characterization of isolates originating from organic chickens is of interest since this type of production might represent another source of exposure of consumers to a variety of the foodborne pathogen C. jejuni. PMID:22468028

  13. Nucleases Encoded by Integrated Elements CJIE2 and CJIE4 Inhibit Natural Transformation of Campylobacter jejuni

    USDA-ARS?s Scientific Manuscript database

    The species Campylobacter jejuni displays huge genetic diversity, and is naturally competent for DNA uptake. Nevertheless, not every strain is able to acquire foreign DNA since nonnaturally transformable strains do exist. Previously we showed that many nonnaturally transformable C. jejuni strains ex...

  14. Differences in the Fecal Concentrations and Genetic Diversities of Campylobacter jejuni Populations among Individual Cows in Two Dairy Herds

    PubMed Central

    Ross, Colleen M.; Pleydell, Eve J.; Muirhead, Richard W.

    2012-01-01

    Dairy cows have been identified as common carriers of Campylobacter jejuni, which causes many of the human gastroenteritis cases reported worldwide. To design on-farm management practices that control the human infection sourced from dairy cows, the first step is to acquire an understanding of the excretion patterns of the cow reservoir. We monitored the same 35 cows from two dairy farms for C. jejuni excretion fortnightly for up to 12 months. The objective was to examine the concentration of C. jejuni and assess the genetic relationship of the C. jejuni populations excreted by individual cows. Significant differences (P < 0.01) in C. jejuni fecal concentration were observed among the 35 cows, with median concentrations that varied by up to 3.6 log10 · g−1 feces. A total of 36 different genotypes were identified from the 514 positive samples by using enterobacterial repetitive intergenic consensus (ERIC)-PCR. Although 22 of these genotypes were excreted by more than one cow, the analysis of frequencies and distribution of the genotypes by model-based statistics revealed a high degree of individuality in the C. jejuni population in each cow. The observed variation in the frequency of excretion of a genotype among cows and the analysis by multilocus sequence typing (MLST) of these genotypes suggest that excretion of C. jejuni in high numbers is due to a successful adaptation of a particular genotype to a particular cow's gut environment, but that animal-related factors render some individual cows resistant to colonization by particular genotypes. The reasons for differences in C. jejuni colonization of animals warrant further investigation. PMID:22904055

  15. Mucosal reactive oxygen species decrease virulence by disrupting Campylobacter jejuni phosphotyrosine signaling

    PubMed Central

    Corcionivoschi, Nicolae; Alvarez, Luis A.; Sharp, Thomas H.; Strengert, Monika; Alemka, Abofu; Mantell, Judith; Verkade, Paul; Knaus, Ulla G.; Bourke, Billy

    2013-01-01

    Summary Reactive oxygen species (ROS) play key roles in mucosal defense, yet how they are induced and the consequences for pathogens are unclear. We report that ROS generated by epithelial NADPH oxidases (Nox1/Duox2) during Campylobacter jejuni infection impair bacterial capsule formation and virulence by altering bacterial signal transduction. Upon C. jejuni invasion, ROS released from the intestinal mucosa inhibit the bacterial phosphotyrosine network that is regulated by the outer membrane tyrosine kinase Cjtk (Cj1170/OMP50). ROS-mediated Cjtk inactivation results in an overall decrease in the phosphorylation of C. jejuni outer membrane / periplasmic proteins including UDP-GlcNAc/Glc 4-epimerase (Gne), an enzyme required for N-glycosylation and capsule formation. Cjtk positively regulates Gne by phosphorylating an active site tyrosine, while loss of Cjtk or ROS treatment inhibits Gne activity, causing altered polysaccharide synthesis. Thus, epithelial NADPH oxidases are an early antibacterial defense system in the intestinal mucosa that modifies virulence by disrupting bacterial signaling. PMID:22817987

  16. The Polysaccharide Capsule of Campylobacter jejuni 81-176 Modulates the Host Immune Response

    DTIC Science & Technology

    2012-12-17

    in C. jejuni colonization of chickens (2, 16). 315 Following restimulation, IL-17 production by CD4+ LPLs was reduced in 316 mice colonized by C...produce anti-bacterial molecules (57). Recently, Th17 350 responses have been demonstrated to have protective roles against Salmonella 351 and...old 498 chickens by Campylobacter jejuni. Appl. Environ. Microbiol. 71:8031-8041. 499 17. Guerry, P., C. M. Szymanski, M. M. Prendergast, T. E. Hickey

  17. Influence of inoculation levels and processing parameters on the survival of Campylobacter jejuni in German style fermented turkey sausages.

    PubMed

    Alter, Thomas; Bori, Anouchka; Hamedi, Ahmad; Ellerbroek, Lüppo; Fehlhaber, Karsten

    2006-10-01

    This study investigated the influence of inoculum levels and manufacturing methods on the survival of Campylobacter (C.) jejuni in raw fermented turkey sausages. Sausages were prepared and inoculated with C. jejuni. After inoculation, these sausages were processed and ripened for 8 days. Samples were taken throughout the ripening process. The presence of C. jejuni was established bacteriologically. Additionally, lactic acid bacteria were enumerated, pH values and water activity were measured to verify the ripening process. To detect changes in genotype and verify the identity of the recovered clones, AFLP analysis was carried out on the re-isolated strains. Whereas no C. jejuni were detectable when inoculating the sausages with the lowest inoculum (0.08-0.44 log(10) cfu/g sausage emulsion), C. jejuni were detectable for 12-24h by enrichment when inoculated with approximately 2 log(10) cfu/g. After inoculation with 4 and 6 log(10) cfu/g respectively, C. jejuni were detectable without enrichment for 12-48 h and by enrichment for 144 h at the most. The greatest decrease of the C. jejuni population occurred during the first 4 h of ripening. Only a very high inoculum level allowed the survival of the organism during a fermentation process and during ripening to pose a potential risk for consumers. Lower initial Campylobacter inoculums will be eliminated during proper ripening of the sausages, if sufficient decrease in water activity and pH-value is ensured.

  18. Comparison of Survival of Campylobacter jejuni in the Phyllosphere with That in the Rhizosphere of Spinach and Radish Plants

    PubMed Central

    Brandl, Maria T.; Haxo, Aileen F.; Bates, Anna H.; Mandrell, Robert E.

    2004-01-01

    Campylobacter jejuni has been isolated previously from market produce and has caused gastroenteritis outbreaks linked to produce. We have tested the ability of this human pathogen to utilize organic compounds that are present in leaf and root exudates and to survive in the plant environment under various conditions. Carbon utilization profiles revealed that C. jejuni can utilize many organic acids and amino acids available on leaves and roots. Despite the presence of suitable substrates in the phyllosphere and the rhizosphere, C. jejuni was unable to grow on lettuce and spinach leaves and on spinach and radish roots of plants incubated at 33°C, a temperature that is conducive to its growth in vitro. However, C. jejuni was cultured from radish roots and from the spinach rhizosphere for at least 23 and 28 days, respectively, at 10°C. This enteric pathogen also persisted in the rhizosphere of spinach for prolonged periods of time at 16°C, a temperature at which many cool-season crops are grown. The decline rate constants of C. jejuni populations in the spinach and radish rhizosphere were 10- and 6-fold lower, respectively, than on healthy spinach leaves at 10°C. The enhanced survival of C. jejuni in soil and in the rhizosphere may be a significant factor in its contamination cycle in the environment and may be associated with the sporadic C. jejuni incidence and campylobacteriosis outbreaks linked to produce. PMID:14766604

  19. Simulation of cross-contamination and decontamination of Campylobacter jejuni during handling of contaminated raw vegetables in a domestic kitchen.

    PubMed

    Chai, Lay-Ching; Lee, Hai-Yen; Ghazali, Farinazleen Mohd; Abu Bakar, Fatimah; Malakar, Pradeep Kumar; Nishibuchi, Mitsuaki; Nakaguchi, Yoshitsugu; Radu, Son

    2008-12-01

    Campylobacter jejuni was found to occur at high prevalence in the raw salad vegetables examined. Previous reports describe cross-contamination involving meat; here we investigated the occurrence of cross-contamination and decontamination events in the domestic kitchen via C. jejuni-contaminated vegetables during salad preparation. This is the first report concerning quantitative cross-contamination and decontamination involving naturally contaminated produce. The study was designed to simulate the real preparation of salad in a household kitchen, starting with washing the vegetables in tap water, then cutting the vegetables on a cutting board, followed by slicing cucumber and blanching (heating in hot water) the vegetables in 85 degrees C water. Vegetables naturally contaminated with C. jejuni were used throughout the simulation to attain realistic quantitative data. The mean of the percent transfer rates for C. jejuni from vegetable to wash water was 30.1 to 38.2%; from wash water to cucumber, it was 26.3 to 47.2%; from vegetables to cutting board, it was 1.6 to 10.3%; and from cutting board to cucumber, it was 22.6 to 73.3%. The data suggest the wash water and plastic cutting board as potential risk factors in C. jejuni transmission to consumers. Washing of the vegetables with tap water caused a 0.4-log reduction of C. jejuni attached to the vegetables (most probable number/gram), while rapid blanching reduced the number of C. jejuni organisms to an undetectable level.

  20. Complete genome sequence of Campylobacter jejuni YH001 from beef liver which contains a novel plasmid

    USDA-ARS?s Scientific Manuscript database

    Campylobacter jejuni is an important foodborne pathogen that causes gastroenteritis in humans and is commonly found in poultry and meat products. Here, we report the complete genome sequence of a Campylobacter jejuni strain recently isolated from retail beef liver. The genome size was 1,712,361 bp, ...

  1. The Evolution of Campylobacter jejuni and Campylobacter coli

    PubMed Central

    Sheppard, Samuel K.; Maiden, Martin C.J.

    2015-01-01

    The global significance of Campylobacter jejuni and Campylobacter coli as gastrointestinal human pathogens has motivated numerous studies to characterize their population biology and evolution. These bacteria are a common component of the intestinal microbiota of numerous bird and mammal species and cause disease in humans, typically via consumption of contaminated meat products, especially poultry meat. Sequence-based molecular typing methods, such as multilocus sequence typing (MLST) and whole genome sequencing (WGS), have been instructive for understanding the epidemiology and evolution of these bacteria and how phenotypic variation relates to the high degree of genetic structuring in C. coli and C. jejuni populations. Here, we describe aspects of the relatively short history of coevolution between humans and pathogenic Campylobacter, by reviewing research investigating how mutation and lateral or horizontal gene transfer (LGT or HGT, respectively) interact to create the observed population structure. These genetic changes occur in a complex fitness landscape with divergent ecologies, including multiple host species, which can lead to rapid adaptation, for example, through frame-shift mutations that alter gene expression or the acquisition of novel genetic elements by HGT. Recombination is a particularly strong evolutionary force in Campylobacter, leading to the emergence of new lineages and even large-scale genome-wide interspecies introgression between C. jejuni and C. coli. The increasing availability of large genome datasets is enhancing understanding of Campylobacter evolution through the application of methods, such as genome-wide association studies, but MLST-derived clonal complex designations remain a useful method for describing population structure. PMID:26101080

  2. Functional and bioinformatics analysis of two Campylobacter jejuni homologs of the thiol-disulfide oxidoreductase, DsbA.

    PubMed

    Grabowska, Anna D; Wywiał, Ewa; Dunin-Horkawicz, Stanislaw; Łasica, Anna M; Wösten, Marc M S M; Nagy-Staroń, Anna; Godlewska, Renata; Bocian-Ostrzycka, Katarzyna; Pieńkowska, Katarzyna; Łaniewski, Paweł; Bujnicki, Janusz M; van Putten, Jos P M; Jagusztyn-Krynicka, E Katarzyna

    2014-01-01

    Bacterial Dsb enzymes are involved in the oxidative folding of many proteins, through the formation of disulfide bonds between their cysteine residues. The Dsb protein network has been well characterized in cells of the model microorganism Escherichia coli. To gain insight into the functioning of the Dsb system in epsilon-Proteobacteria, where it plays an important role in the colonization process, we studied two homologs of the main Escherichia coli Dsb oxidase (EcDsbA) that are present in the cells of the enteric pathogen Campylobacter jejuni, the most frequently reported bacterial cause of human enteritis in the world. Phylogenetic analysis suggests the horizontal transfer of the epsilon-Proteobacterial DsbAs from a common ancestor to gamma-Proteobacteria, which then gave rise to the DsbL lineage. Phenotype and enzymatic assays suggest that the two C. jejuni DsbAs play different roles in bacterial cells and have divergent substrate spectra. CjDsbA1 is essential for the motility and autoagglutination phenotypes, while CjDsbA2 has no impact on those processes. CjDsbA1 plays a critical role in the oxidative folding that ensures the activity of alkaline phosphatase CjPhoX, whereas CjDsbA2 is crucial for the activity of arylsulfotransferase CjAstA, encoded within the dsbA2-dsbB-astA operon. Our results show that CjDsbA1 is the primary thiol-oxidoreductase affecting life processes associated with bacterial spread and host colonization, as well as ensuring the oxidative folding of particular protein substrates. In contrast, CjDsbA2 activity does not affect the same processes and so far its oxidative folding activity has been demonstrated for one substrate, arylsulfotransferase CjAstA. The results suggest the cooperation between CjDsbA2 and CjDsbB. In the case of the CjDsbA1, this cooperation is not exclusive and there is probably another protein to be identified in C. jejuni cells that acts to re-oxidize CjDsbA1. Altogether the data presented here constitute the

  3. Functional and Bioinformatics Analysis of Two Campylobacter jejuni Homologs of the Thiol-Disulfide Oxidoreductase, DsbA

    PubMed Central

    Grabowska, Anna D.; Wywiał, Ewa; Dunin-Horkawicz, Stanislaw; Łasica, Anna M.; Wösten, Marc M. S. M.; Nagy-Staroń, Anna; Godlewska, Renata; Bocian-Ostrzycka, Katarzyna; Pieńkowska, Katarzyna; Łaniewski, Paweł; Bujnicki, Janusz M.; van Putten, Jos P. M.; Jagusztyn-Krynicka, E. Katarzyna

    2014-01-01

    Background Bacterial Dsb enzymes are involved in the oxidative folding of many proteins, through the formation of disulfide bonds between their cysteine residues. The Dsb protein network has been well characterized in cells of the model microorganism Escherichia coli. To gain insight into the functioning of the Dsb system in epsilon-Proteobacteria, where it plays an important role in the colonization process, we studied two homologs of the main Escherichia coli Dsb oxidase (EcDsbA) that are present in the cells of the enteric pathogen Campylobacter jejuni, the most frequently reported bacterial cause of human enteritis in the world. Methods and Results Phylogenetic analysis suggests the horizontal transfer of the epsilon-Proteobacterial DsbAs from a common ancestor to gamma-Proteobacteria, which then gave rise to the DsbL lineage. Phenotype and enzymatic assays suggest that the two C. jejuni DsbAs play different roles in bacterial cells and have divergent substrate spectra. CjDsbA1 is essential for the motility and autoagglutination phenotypes, while CjDsbA2 has no impact on those processes. CjDsbA1 plays a critical role in the oxidative folding that ensures the activity of alkaline phosphatase CjPhoX, whereas CjDsbA2 is crucial for the activity of arylsulfotransferase CjAstA, encoded within the dsbA2-dsbB-astA operon. Conclusions Our results show that CjDsbA1 is the primary thiol-oxidoreductase affecting life processes associated with bacterial spread and host colonization, as well as ensuring the oxidative folding of particular protein substrates. In contrast, CjDsbA2 activity does not affect the same processes and so far its oxidative folding activity has been demonstrated for one substrate, arylsulfotransferase CjAstA. The results suggest the cooperation between CjDsbA2 and CjDsbB. In the case of the CjDsbA1, this cooperation is not exclusive and there is probably another protein to be identified in C. jejuni cells that acts to re-oxidize CjDsbA1. Altogether

  4. Examination of Campylobacter jejuni putative adhesins leads to the identification of a new protein, designated FlpA, required for chicken colonization

    USDA-ARS?s Scientific Manuscript database

    Campylobacter jejuni colonization of chickens is dependent upon surface exposed proteins termed adhesins. Putative C. jejuni adhesins include CadF, CapA, JlpA, MOMP, PEB1, Cj1279c, and Cj1349c. We examined the genetic relatedness of ninety-seven C. jejuni isolates recovered from human, poultry, bo...

  5. Boiling incipience and convective boiling of neon and nitrogen

    NASA Technical Reports Server (NTRS)

    Papell, S. S.; Hendricks, R. C.

    1977-01-01

    Forced convection and subcooled boiling heat transfer data for liquid nitrogen and liquid neon were obtained in support of a design study for a 30 tesla cryomagnet cooled by forced convection of liquid neon. The cryogen data obtained over a range of system pressures, fluid flow rates, and applied heat fluxes were used to develop correlations for predicting boiling incipience and convective boiling heat transfer coefficients in uniformly heated flow channels. The accuracy of the correlating equations was then evaluated. A technique was also developed to calculate the position of boiling incipience in a uniformly heated flow channel. Comparisons made with the experimental data showed a prediction accuracy of + or - 15 percent.

  6. Rapid identification and quantification of Campylobacter coli and Campylobacter jejuni by real-time PCR in pure cultures and in complex samples

    PubMed Central

    2011-01-01

    Background Campylobacter spp., especially Campylobacter jejuni (C. jejuni) and Campylobacter coli (C. coli), are recognized as the leading human foodborne pathogens in developed countries. Livestock animals carrying Campylobacter pose an important risk for human contamination. Pigs are known to be frequently colonized with Campylobacter, especially C. coli, and to excrete high numbers of this pathogen in their faeces. Molecular tools, notably real-time PCR, provide an effective, rapid, and sensitive alternative to culture-based methods for the detection of C. coli and C. jejuni in various substrates. In order to serve as a diagnostic tool supporting Campylobacter epidemiology, we developed a quantitative real-time PCR method for species-specific detection and quantification of C. coli and C. jejuni directly in faecal, feed, and environmental samples. Results With a sensitivity of 10 genome copies and a linear range of seven to eight orders of magnitude, the C. coli and C. jejuni real-time PCR assays allowed a precise quantification of purified DNA from C. coli and C. jejuni. The assays were highly specific and showed a 6-log-linear dynamic range of quantification with a quantitative detection limit of approximately 2.5 × 102 CFU/g of faeces, 1.3 × 102 CFU/g of feed, and 1.0 × 103 CFU/m2 for the environmental samples. Compared to the results obtained by culture, both C. coli and C. jejuni real-time PCR assays exhibited a specificity of 96.2% with a kappa of 0.94 and 0.89 respectively. For faecal samples of experimentally infected pigs, the coefficients of correlation between the C. coli or C. jejuni real-time PCR assay and culture enumeration were R2 = 0.90 and R2 = 0.93 respectively. Conclusion The C. coli and C. jejuni real-time quantitative PCR assays developed in this study provide a method capable of directly detecting and quantifying C. coli and C. jejuni in faeces, feed, and environmental samples. These assays represent a new diagnostic tool for studying

  7. The significance of Campylobacter jejuni infection in poultry: a review.

    PubMed

    Shane, S M

    1992-01-01

    Campylobacter is a significant cause of enterocolitis in consumers of undercooked poultry meat. Campylobacter jejuni is the most significant of the three thermophilic Campylobacter species, and is responsible for intestinal colonization in poultry and food-borne enteritis in humans. Generally, C. jejuni is apathogenic in poultry, although newly hatched chicks and turkeys may develop a transient diarrhoea following infection. Modern intensive poultry production favours the introduction of infection into commercial growing units, resulting in intestinal colonization during the second to fourth weeks inclusive. Routes of infection include contaminated fomites, infected water supply, rodents, insects, and free-living birds. Vertical transmission is considered unlikely. Contamination of poultry meat is enhanced by deficiencies in transport and processing of broilers and turkeys. Scalding, defeathering and evisceration represent the significant points of cross-contamination during processing. Epidemiological correlation has been established between consumption of contaminated chicken and outbreaks of human campylobacteriosis. Amelioration of infection by application of improved standards of hygiene and decontamination is possible in the context of commercial poultry production. Improvement in washing of carcasses, and the application of chemical disinfectants and gamma irradiation have the potential to reduce the prevalence of C. jejuni contamination in poultry meat. These innovations, together with improved storage and handling of meat products, will reduce the risk of campylobacteriosis to consumers.

  8. Biosynthesis of Nucleoside Diphosphoramidates in Campylobacter jejuni.

    PubMed

    Taylor, Zane W; Brown, Haley A; Holden, Hazel M; Raushel, Frank M

    2017-11-21

    Campylobacter jejuni is a pathogenic Gram-negative bacterium and a leading cause of food-borne gastroenteritis. C. jejuni produces a capsular polysaccharide (CPS) that contains a unique O-methyl phosphoramidate modification (MeOPN). Recently, the first step in the biosynthetic pathway for the assembly of the MeOPN modification to the CPS was elucidated. It was shown that the enzyme Cj1418 catalyzes the phosphorylation of the amide nitrogen of l-glutamine to form l-glutamine phosphate. In this investigation, the metabolic fate of l-glutamine phosphate was determined. The enzyme Cj1416 catalyzes the displacement of pyrophosphate from MgCTP by l-glutamine phosphate to form CDP-l-glutamine. The enzyme Cj1417 subsequently catalyzes the hydrolysis of CDP-l-glutamine to generate cytidine diphosphoramidate and l-glutamate. The structures of the two novel intermediates, CDP-l-glutamine and cytidine diphosphoramidate, were confirmed by 31 P nuclear magnetic resonance spectroscopy and mass spectrometry. It is proposed that the enzyme Cj1416 be named CTP:phosphoglutamine cytidylyltransferase and that the enzyme Cj1417 be named γ-glutamyl-CDP-amidate hydrolase.

  9. The complete annotated genome sequences of three Campylobacter jejuni strains isolated from naturally colonized, farm raised chickens

    USDA-ARS?s Scientific Manuscript database

    Campylobacter jejuni is a leading cause of bacterially derived foodborne illness worldwide. Human illness is commonly associated with handling and consumption of contaminated poultry products. Three C. jejuni strains were isolated from cecal contents of three different naturally colonized, farm rais...

  10. Complete genome sequence of Campylobacter jejuni RM1246-ERRC that exhibits resistance to Quaternary Ammonium Compounds

    USDA-ARS?s Scientific Manuscript database

    Campylobacter jejuni strain RM1246-ERRC is a clinical isolate. In laboratory experiments RM1246-ERRC exhibited resistance to the antimicrobial effects of quaternary ammonium compounds (QACs) when compared to other C. jejuni strains. The chromosome of RM1246-ERRC was determined to be 1,659,694 bp w...

  11. Prevalence, antibiogram, and cdt genes of toxigenic Campylobacter jejuni in salad style vegetables (ulam) at farms and retail outlets in Terengganu.

    PubMed

    Khalid, Mohd Ikhsan; Tang, John Yew Huat; Baharuddin, Nabila Huda; Rahman, Nasiha Shakina; Rahimi, Nurul Faizzah; Radu, Son

    2015-01-01

    The present study was conducted to investigate the prevalence and antibiotic resistance among Campylobacter jejuni in ulam at farms and retail outlets located in Kuala Terengganu, Malaysia. A total of 526 samples (ulam, soil, and fertilizer) were investigated for the presence of C. jejuni and the gene for cytolethal distending toxin (cdt) by using a multiplex PCR method. Antibiotic susceptibility to 10 types of antibiotics was determined using the disk diffusion method for 33 C. jejuni isolates. The average prevalence of contaminated samples from farms, wet markets, and supermarkets was 35.29, 52.66, and 69.88%, respectively. The cdt gene was not detected in 24 of the 33 C. jejuni isolates, but 9 isolates harbored cdtC. Antibiotic resistance in C. jejuni isolates was highest to penicillin G (96.97% of isolates) followed by vancomycin (87.88%), ampicillin (75.76%), erythromycin (60.61%), tetracycline (9.09%), amikacin (6.06%), and norfloxacin (3.03%); none of the isolates were resistant to ciprofloxacin, enrofloxacin, and gentamicin. In this study, C. jejuni was present in ulam, and some isolates were highly resistant to some antibiotics but not to quinolones. Thus, appropriate attention and measures are required to prevent C. jejuni contamination on farms and at retail outlets.

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

    PubMed

    Johansen, C H; Bjerrum, L; Finster, K; Pedersen, K

    2006-04-01

    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 effect of a C. jejuni infection on the ileal microflora of the broilers. In addition, denaturant gradient gel electrophoresis (DGGE) profiles generated from cecal and ileal contents revealed several DGGE bands that were present in the control chickens, but not in the chickens colonized with C. jejuni. Some of these DGGE bands could be affiliated with Lactobacillus reuteri, Clostridium perfringens, and the genus Klebsiella.

  13. Antimicrobial resistance of Campylobacter jejuni and Campylobacter coli from poultry in Italy.

    PubMed

    Giacomelli, Martina; Salata, Cristiano; Martini, Marco; Montesissa, Clara; Piccirillo, Alessandra

    2014-04-01

    This study was aimed at assessing the antimicrobial resistance (AMR) of Campylobacter isolates from broilers and turkeys reared in industrial farms in Northern Italy, given the public health concern represented by resistant campylobacters in food-producing animals and the paucity of data about this topic in our country. Thirty-six Campylobacter jejuni and 24 Campylobacter coli isolated from broilers and 68 C. jejuni and 32 C. coli from turkeys were tested by disk diffusion for their susceptibility to apramycin, gentamicin, streptomycin, cephalothin, cefotaxime, ceftiofur, cefuroxime, ampicillin, amoxicillin+clavulanic acid, nalidixic acid, flumequine, enrofloxacin, ciprofloxacin, erythromycin, tilmicosin, tylosin, tiamulin, clindamycin, tetracycline, sulfamethoxazole+trimethoprim, chloramphenicol. Depending on the drug, breakpoints provided by Comité de l'antibiogramme de la Société Française de Microbiologie, Clinical and Laboratory Standards Institute, and the manufacturer were followed. All broiler strains and 92% turkey strains were multidrug resistant. Very high resistance rates were detected for quinolones, tetracycline, and sulfamethoxazole+trimethoprim, ranging from 65% to 100% in broilers and from 74% to 96% in turkeys. Prevalence of resistance was observed also against ampicillin (97% in broilers, 88% in turkeys) and at least three cephalosporins (93-100% in broilers, 100% in turkeys). Conversely, no isolates showed resistance to chloramphenicol and tiamulin. Susceptibility prevailed for amoxicillin+clavulanic acid and aminoglycosides in both poultry species, and for macrolides and clindamycin among turkey strains and among C. jejuni from broilers, whereas most C. coli strains from broilers (87.5%) were resistant. Other differences between C. jejuni and C. coli were observed markedly in broiler isolates, with the overall predominance of resistance in C. coli compared to C. jejuni. This study provides updates and novel data on the AMR of broiler and

  14. In vivo and in silico determination of essential genes of Campylobacter jejuni.

    PubMed

    Metris, Aline; Reuter, Mark; Gaskin, Duncan J H; Baranyi, Jozsef; van Vliet, Arnoud H M

    2011-11-01

    In the United Kingdom, the thermophilic Campylobacter species C. jejuni and C. coli are the most frequent causes of food-borne gastroenteritis in humans. While campylobacteriosis is usually a relatively mild infection, it has a significant public health and economic impact, and possible complications include reactive arthritis and the autoimmune diseases Guillain-Barré syndrome. The rapid developments in "omics" technologies have resulted in the availability of diverse datasets allowing predictions of metabolism and physiology of pathogenic micro-organisms. When combined, these datasets may allow for the identification of potential weaknesses that can be used for development of new antimicrobials to reduce or eliminate C. jejuni and C. coli from the food chain. A metabolic model of C. jejuni was constructed using the annotation of the NCTC 11168 genome sequence, a published model of the related bacterium Helicobacter pylori, and extensive literature mining. Using this model, we have used in silico Flux Balance Analysis (FBA) to determine key metabolic routes that are essential for generating energy and biomass, thus creating a list of genes potentially essential for growth under laboratory conditions. To complement this in silico approach, candidate essential genes have been determined using a whole genome transposon mutagenesis method. FBA and transposon mutagenesis (both this study and a published study) predict a similar number of essential genes (around 200). The analysis of the intersection between the three approaches highlights the shikimate pathway where genes are predicted to be essential by one or more method, and tend to be network hubs, based on a previously published Campylobacter protein-protein interaction network, and could therefore be targets for novel antimicrobial therapy. We have constructed the first curated metabolic model for the food-borne pathogen Campylobacter jejuni and have presented the resulting metabolic insights. We have shown that

  15. Microbiota-derived short-chain fatty acids modulate expression of Campylobacter jejuni determinants required for commensalism and virulence

    USDA-ARS?s Scientific Manuscript database

    Campylobacter jejuni effectively promotes commensalism in the intestinal tract of avian hosts and diarrheal disease in humans, yet components of intestinal environments sensed by the bacterium in either host to initiate interactions are mostly unknown. By analyzing a C. jejuni acetogenesis mutant th...

  16. Secondary pool boiling effects

    NASA Astrophysics Data System (ADS)

    Kruse, C.; Tsubaki, A.; Zuhlke, C.; Anderson, T.; Alexander, D.; Gogos, G.; Ndao, S.

    2016-02-01

    A pool boiling phenomenon referred to as secondary boiling effects is discussed. Based on the experimental trends, a mechanism is proposed that identifies the parameters that lead to this phenomenon. Secondary boiling effects refer to a distinct decrease in the wall superheat temperature near the critical heat flux due to a significant increase in the heat transfer coefficient. Recent pool boiling heat transfer experiments using femtosecond laser processed Inconel, stainless steel, and copper multiscale surfaces consistently displayed secondary boiling effects, which were found to be a result of both temperature drop along the microstructures and nucleation characteristic length scales. The temperature drop is a function of microstructure height and thermal conductivity. An increased microstructure height and a decreased thermal conductivity result in a significant temperature drop along the microstructures. This temperature drop becomes more pronounced at higher heat fluxes and along with the right nucleation characteristic length scales results in a change of the boiling dynamics. Nucleation spreads from the bottom of the microstructure valleys to the top of the microstructures, resulting in a decreased surface superheat with an increasing heat flux. This decrease in the wall superheat at higher heat fluxes is reflected by a "hook back" of the traditional boiling curve and is thus referred to as secondary boiling effects. In addition, a boiling hysteresis during increasing and decreasing heat flux develops due to the secondary boiling effects. This hysteresis further validates the existence of secondary boiling effects.

  17. Flagella-Mediated Adhesion and Extracellular DNA Release Contribute to Biofilm Formation and Stress Tolerance of Campylobacter jejuni

    PubMed Central

    Svensson, Sarah L.; Pryjma, Mark; Gaynor, Erin C.

    2014-01-01

    Campylobacter jejuni is a leading cause of foodbourne gastroenteritis, despite fragile behaviour under standard laboratory conditions. In the environment, C. jejuni may survive within biofilms, which can impart resident bacteria with enhanced stress tolerance compared to their planktonic counterparts. While C. jejuni forms biofilms in vitro and in the wild, it had not been confirmed that this lifestyle confers stress tolerance. Moreover, little is understood about molecular mechanisms of biofilm formation in this pathogen. We previously found that a ΔcprS mutant, which carries a deletion in the sensor kinase of the CprRS two-component system, forms enhanced biofilms. Biofilms were also enhanced by the bile salt deoxycholate and contained extracellular DNA. Through more in-depth analysis of ΔcprS and WT under conditions that promote or inhibit biofilms, we sought to further define this lifestyle for C. jejuni. Epistasis experiments with ΔcprS and flagellar mutations (ΔflhA, ΔpflA) suggested that initiation is mediated by flagellum-mediated adherence, a process which was kinetically enhanced by motility. Lysis was also observed, especially under biofilm-enhancing conditions. Microscopy suggested adherence was followed by release of eDNA, which was required for biofilm maturation. Importantly, inhibiting biofilm formation by removal of eDNA with DNase decreased stress tolerance. This work suggests the biofilm lifestyle provides C. jejuni with resilience that has not been apparent from observation of planktonic bacteria during routine laboratory culture, and provides a framework for subsequent molecular studies of C. jejuni biofilms. PMID:25166748

  18. Identification of a Novel Membrane Transporter Mediating Resistance to Organic Arsenic in Campylobacter jejuni

    PubMed Central

    Shen, Zhangqi; Luangtongkum, Taradon; Qiang, Zhiyi; Jeon, Byeonghwa; Wang, Liping

    2014-01-01

    Although bacterial mechanisms involved in the resistance to inorganic arsenic are well understood, the molecular basis for organic arsenic resistance has not been described. Campylobacter jejuni, a major food-borne pathogen causing gastroenteritis in humans, is highly prevalent in poultry and is reportedly resistant to the arsenic compound roxarsone (4-hydroxy-3-nitrobenzenearsonic acid), which has been used as a feed additive in the poultry industry for growth promotion. In this study, we report the identification of a novel membrane transporter (named ArsP) that contributes to organic arsenic resistance in Campylobacter. ArsP is predicted to be a membrane permease containing eight transmembrane helices, distinct from other known arsenic transporters. Analysis of multiple C. jejuni isolates from various animal species revealed that the presence of an intact arsP gene is associated with elevated resistance to roxarsone. In addition, inactivation of arsP in C. jejuni resulted in 4- and 8-fold reductions in the MICs of roxarsone and nitarsone, respectively, compared to that for the wild-type strain. Furthermore, cloning of arsP into a C. jejuni strain lacking a functional arsP gene led to 16- and 64-fold increases in the MICs of roxarsone and nitarsone, respectively. Neither mutation nor overexpression of arsP affected the MICs of inorganic arsenic, including arsenite and arsenate, in Campylobacter. Moreover, acquisition of arsP in NCTC 11168 led to accumulation of less roxarsone than the wild-type strain lacking arsP. Together, these results indicate that ArsP functions as an efflux transporter specific for extrusion of organic arsenic and contributes to the resistance to these compounds in C. jejuni. PMID:24419344

  19. Development of an Immunoassay for Rapid Detection of Ganglioside GM1 Mimicry in Campylobacter jejuni Strains

    PubMed Central

    Prendergast, Martina M.; Kosunen, Timo U.; Moran, Anthony P.

    2001-01-01

    Mimicry of peripheral nerve gangliosides by Campylobacter jejuni lipopolysaccharides (LPSs) has been proposed to induce cross-reacting antiganglioside antibodies in Guillain-Barré syndrome (GBS). Because current methods for LPS characterization are labor-intensive and inhibit the screening of large numbers of strains, a rapid GM1 epitope screening assay was developed. Biomass from two agar plates of confluent growth yielded sufficient LPS using a novel phenol-water and ether extraction procedure. Extracts of LPS were reacted with cholera toxin (GM1 ligand), peanut agglutinin (Galβ1→3GalNAc ligand), and anti-GM1 antibodies. After the assay was validated, 12 of 59 (20%) C. jejuni serostrains, including four serotypes that have not previously been associated with GBS, reacted with two or more anti-GM1 ganglioside reagents. Subsequently, LPS extracts from 5 of 7 (71%) C. jejuni isolates and 2 of 3 (67%) C. jejuni culture collection strains bore GM1 structures. Overall, the assay system was reliable, efficient, and reproducible and may be adapted for large-scale epidemiological studies. PMID:11283076

  20. The antimicrobial effect of spice-based marinades against Campylobacter jejuni on contaminated fresh broiler wings.

    PubMed

    Zakarienė, Gintarė; Rokaitytė, Anita; Ramonaitė, Sigita; Novoslavskij, Aleksandr; Mulkytė, Kristina; Zaborskienė, Gintarė; Malakauskas, Mindaugas

    2015-03-01

    The antimicrobial effect of spice-based marinades against Campylobacter jejuni on inoculated fresh broiler wings was investigated. Experiments were carried out with 1 strain of C. jejuni and 6 marinades. Four experimental marinades were composed for the study and contained spices (thyme, rosemary, basil, marjoram, and so on) and different combination of bioactive compounds. Two marinades were commercial and contained spices (black pepper, sweet red pepper, and so on) and chemical additives (monosodium glutamate, sodium diacetate, calcium lactate), 1 commercial marinade was also enriched with bioactive compounds (linalool, cinnamaldehyde, lactic acid). Total aerobic bacterial count was examined to estimate the possible effect of tested marinades on the shelf-life of marinated broiler wings. Study revealed that thyme-based marinade was the most effective against C. jejuni on broiler wings and reduced the numbers of campylobacters by 1.04 log colony forming unit (CFU)/g (P ≤ 0.05) during storage for 168 h at 4 °C temperature. Moreover, it was more effective against C. jejuni than commercial marinade with 0.47 log CFU/g (P ≤ 0.05) reduction effect. Both experimental and commercial marinades had very similar effect on the total aerobic bacterial count. Although experimental and commercial marinades had different effect on pH of broiler wings, this parameter did not show a major impact on the antimicrobial effect of tested marinades (P ≥ 0.05). Our study shows that experimental natural thyme-based marinade can reduce numbers of C. jejuni more effectively than tested commercial marinades. © 2015 Institute of Food Technologists®

  1. When water does not boil at the boiling point.

    PubMed

    Chang, Hasok

    2007-03-01

    Every schoolchild learns that, under standard pressure, pure water always boils at 100 degrees C. Except that it does not. By the late 18th century, pioneering scientists had already discovered great variations in the boiling temperature of water under fixed pressure. So, why have most of us been taught that the boiling point of water is constant? And, if it is not constant, how can it be used as a 'fixed point' for the calibration of thermometers? History of science has the answers.

  2. PFGE, Lior serotype, and antimicrobial resistance patterns among Campylobacter jejuni isolated from travelers and US military personnel with acute diarrhea in Thailand, 1998-2003

    PubMed Central

    2010-01-01

    Background Campylobacter jejuni is a major cause of gastroenteritis worldwide. In Thailand, several strains of C. jejuni have been isolated and identified as major diarrheal pathogens among adult travelers. To study the epidemiology of C. jejuni in adult travelers and U.S. military personnel with acute diarrhea in Thailand from 1998-2003, strains of C. jejuni were isolated and phenotypically identified, serotyped, tested for antimicrobial susceptibility, and characterized using pulsed-field gel electrophoresis (PFGE). Results A total of 312 C. jejuni isolates were obtained from travelers (n = 46) and U.S. military personnel (n = 266) in Thailand who were experiencing acute diarrhea. Nalidixic acid and ciprofloxacin resistance was observed in 94.9% and 93.0% of the isolates, respectively. From 2001-2003, resistance to tetracycline (81.9%), trimethoprim-sulfamethoxazole (57.9%), ampicillin (28.9%), kanamycin (5.9%), sulfisoxazole (3.9%), neomycin (2.0%), and streptomycin (0.7%) was observed. Combined PFGE analysis showed considerable genetic diversity among the C. jejuni isolates; however, four PFGE clusters included isolates from the major Lior serotypes (HL: 36, HL: 11, HL: 5, and HL: 28). The PFGE analysis linked individual C. jejuni clones that were obtained at U.S. military exercises with specific antimicrobial resistance patterns. Conclusions In summary, most human C. jejuni isolates from Thailand were multi-resistant to quinolones and tetracycline. PFGE detected spatial and temporal C. jejuni clonality responsible for the common sources of Campylobacter gastroenteritis. PMID:21062505

  3. RNAseq Reveals Complex Response of Campylobacter jejuni to Ovine Bile and In vivo Gallbladder Environment

    PubMed Central

    Kreuder, Amanda J.; Schleining, Jennifer A.; Yaeger, Michael; Zhang, Qijing; Plummer, Paul J.

    2017-01-01

    Colonization of the gallbladder by enteric pathogens such as Salmonella typhi, Listeria monocytogenes, and Campylobacter jejuni is thought to play a key role in transmission and persistence of these important zoonotic agents; however, little is known about the molecular mechanisms that allow for bacterial survival within this harsh environment. Recently, a highly virulent C. jejuni sheep abortion (SA) clone represented by the clinical isolate IA3902 has emerged as the dominant cause for sheep abortion in the United States. Previous studies have indicated that the C. jejuni clone SA can frequently be isolated from the gallbladders of otherwise healthy sheep, suggesting that the gallbladder may serve as an important reservoir for infection. To begin to understand the molecular mechanisms associated with survival in the host gallbladder, C. jejuni IA3902 was exposed for up to 24 h to both the natural ovine host in vivo gallbladder environment, as well as ovine bile in vitro. Following exposure, total RNA was isolated from the bile and high throughput deep sequencing of strand specific rRNA-depleted total RNA was used to characterize the transcriptome of IA3902 under these conditions. Our results demonstrated for the first time the complete transcriptome of C. jejuni IA3902 during exposure to an important host environment, the sheep gallbladder. Exposure to the host environment as compared to in vitro bile alone provided a more robust picture of the complexity of gene regulation required for survival in the host gallbladder. A subset of genes including a large number of protein coding genes as well as seven previously identified non-coding RNAs were confirmed to be differentially expressed within our data, suggesting that they may play a key role in adaptation upon exposure to these conditions. This research provides valuable insights into the molecular mechanisms that may be utilized by C. jejuni IA3902 to colonize and survive within the inhospitable gallbladder

  4. Structures and transport dynamics of a Campylobacter jejuni multidrug efflux pump

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Su, Chih-Chia; Yin, Linxiang; Kumar, Nitin

    2017-08-01

    Resistance-nodulation-cell division efflux pumps are integral membrane proteins that catalyze the export of substrates across cell membranes. Within the hydrophobe-amphiphile efflux subfamily, these resistance-nodulation-cell division proteins largely form trimeric efflux pumps. The drug efflux process has been proposed to entail a synchronized motion between subunits of the trimer to advance the transport cycle, leading to the extrusion of drug molecules. Here we use X-ray crystallography and single-molecule fluorescence resonance energy transfer imaging to elucidate the structures and functional dynamics of the Campylobacter jejuni CmeB multidrug efflux pump. We find that the CmeB trimer displays a very unique conformation. A directmore » observation of transport dynamics in individual CmeB trimers embedded in membrane vesicles indicates that each CmeB subunit undergoes conformational transitions uncoordinated and independent of each other. On the basis of our findings and analyses, we propose a model for transport mechanism where CmeB protomers function independently within the trimer.« less

  5. Effect of refrigeration and frozen storage on the Campylobacter jejuni recovery from naturally contaminated broiler carcasses

    PubMed Central

    Maziero, Maike T.; de Oliveira, Tereza Cristina R. M.

    2010-01-01

    Campylobacter jejuni is the most common thermophilic Campylobacter associated with human enteritis in many countries. Broilers and their by-products are the main sources for human enteritis. Refrigeration and freezing are used to control bacterial growth in foods. The effect of these interventions on survival of Campylobacter jejuni is yet not quite understood. This study evaluated the effect of storage temperature on the survival of C. jejuni in chicken meat stored for seven days at 4°C and for 28 days at -20°C. The influence of selective enrichment on recovery of Campylobacter was also evaluated. Thirty fresh chicken meat samples were analyzed and 93.3% was contaminated with termotolerant Campylobacter spp. with average count of 3.08 Log10 CFU/g on direct plating. After refrigeration, 53.3% of the analyzed samples tested positive for Campylobacter and the average count was 1.19 Log10 CFU/g. After storage at -20°C, 36.6% of the samples were positive with a verage count of 0.75 Log10 CFU/g. C. jejuni was detected after enrichment, respectively, in 50% of the fresh, 36.7% of the refrigerated and 33.3% of the frozen meat samples analyzed. No difference was detected for the recovery of C. jejuni from fresh, refrigerated or frozen samples after selective enrichment, showing that this microorganism can survive under the tested storage conditions. PMID:24031523

  6. Draft Genome Sequences of 116 Campylobacter jejuni Strains Isolated from Humans, Animals, Food, and the Environment in Brazil.

    PubMed

    Frazão, Miliane Rodrigues; Cao, Guojie; Medeiros, Marta Inês Cazentini; Duque, Sheila da Silva; Leon, Maria Sanchez; Allard, Marc William; Falcão, Juliana Pfrimer

    2018-04-19

    Campylobacter jejuni is a major zoonotic pathogen that causes foodborne gastroenteritis worldwide. However, clinical cases of campylobacteriosis have been underreported and underdiagnosed in Brazil. Herein, we describe the draft genome sequences of 116 C. jejuni strains isolated from diverse sources in Brazil.

  7. Lattice Boltzmann modeling of boiling heat transfer: The boiling curve and the effects of wettability

    DOE PAGES

    Li, Q.; Kang, Q. J.; Francois, M. M.; ...

    2015-03-03

    A hybrid thermal lattice Boltzmann (LB) model is presented to simulate thermal multiphase flows with phase change based on an improved pseudopotential LB approach (Li et al., 2013). The present model does not suffer from the spurious term caused by the forcing-term effect, which was encountered in some previous thermal LB models for liquid–vapor phase change. Using the model, the liquid–vapor boiling process is simulated. The boiling curve together with the three boiling stages (nucleate boiling, transition boiling, and film boiling) is numerically reproduced in the LB community for the first time. The numerical results show that the basic featuresmore » and the fundamental characteristics of boiling heat transfer are well captured, such as the severe fluctuation of transient heat flux in the transition boiling and the feature that the maximum heat transfer coefficient lies at a lower wall superheat than that of the maximum heat flux. Moreover, the effects of the heating surface wettability on boiling heat transfer are investigated. It is found that an increase in contact angle promotes the onset of boiling but reduces the critical heat flux, and makes the boiling process enter into the film boiling regime at a lower wall superheat, which is consistent with the findings from experimental studies.« less

  8. Standing Genetic Variation in Contingency Loci Drives the Rapid Adaptation of Campylobacter jejuni to a Novel Host

    PubMed Central

    Jerome, John P.; Bell, Julia A.; Plovanich-Jones, Anne E.; Barrick, Jeffrey E.; Brown, C. Titus; Mansfield, Linda S.

    2011-01-01

    The genome of the food-borne pathogen Campylobacter jejuni contains multiple highly mutable sites, or contingency loci. It has been suggested that standing variation at these loci is a mechanism for rapid adaptation to a novel environment, but this phenomenon has not been shown experimentally. In previous work we showed that the virulence of C. jejuni NCTC11168 increased after serial passage through a C57BL/6 IL-10-/- mouse model of campylobacteriosis. Here we sought to determine the genetic basis of this adaptation during passage. Re-sequencing of the 1.64Mb genome to 200-500X coverage allowed us to define variation in 23 contingency loci to an unprecedented depth both before and after in vivo adaptation. Mutations in the mouse-adapted C. jejuni were largely restricted to the homopolymeric tracts of thirteen contingency loci. These changes cause significant alterations in open reading frames of genes in surface structure biosynthesis loci and in genes with only putative functions. Several loci with open reading frame changes also had altered transcript abundance. The increase in specific phases of contingency loci during in vivo passage of C. jejuni, coupled with the observed virulence increase and the lack of other types of genetic changes, is the first experimental evidence that these variable regions play a significant role in C. jejuni adaptation and virulence in a novel host. PMID:21283682

  9. Monomorphic genotypes within a generalist lineage of Campylobacter jejuni show signs of global dispersion

    PubMed Central

    Zhang, Ji; Vehkala, Minna; Välimäki, Niko; Hakkinen, Marjaana; Hänninen, Marja-Liisa; Roasto, Mati; Mäesaar, Mihkel; Taboada, Eduardo; Barker, Dillon; Garofolo, Giuliano; Cammà, Cesare; Di Giannatale, Elisabetta; Corander, Jukka; Rossi, Mirko

    2016-01-01

    The decreased costs of genome sequencing have increased the capability to apply whole-genome sequencing to epidemiological surveillance of zoonotic Campylobacter jejuni. However, knowledge of the genetic diversity of this bacteria is vital for inferring relatedness between epidemiologically linked isolates and a necessary prerequisite for correct application of this methodology. To address this issue in C. jejuni we investigated the spatial and temporal signals in the genomes of a major clonal complex and generalist lineage, ST-45 CC, by analysing the population structure and genealogy as well as applying genome-wide association analysis of 340 isolates from across Europe collected over a wide time range. The occurrence and strength of the geographical signal varied between sublineages and followed the clonal frame when present, while no evidence of a temporal signal was found. Certain sublineages of ST-45 formed discrete and genetically isolated clades containing isolates with extremely similar genomes regardless of time and location of sampling. Based on a separate data set, these monomorphic genotypes represent successful C. jejuni clones, possibly spread around the globe by rapid animal (migrating birds), food or human movement. In addition, we observed an incongruence between the genealogy of the strains and multilocus sequence typing (MLST), challenging the existing clonal complex definition and the use of whole-genome gene-by-gene hierarchical nomenclature schemes for C. jejuni. PMID:28348829

  10. Detection of Campylobacter jejuni added to foods by using a combined selective enrichment and nucleic acid sequence-based amplification (NASBA).

    PubMed Central

    Uyttendaele, M; Schukkink, R; van Gemen, B; Debevere, J

    1995-01-01

    An assay to detect Campylobacter jejuni in foods that uses a short selective enrichment culture, a simple and rapid isolation procedure, NASBA amplification of RNA, and a nonradioactive in solution hybridization was studied. The presence of high numbers of indigenous flora affected the sensitivity of the assay. However, detection of C. jejuni was possible up to a ratio of indigenous flora to C. jejuni of 10,000:1. Interference by food components was eliminated by centrifugation following the enrichment step. Fourteen food samples artificially inoculated with C. jejuni (1 to 1,000 CFU/10 g) were analyzed with the NASBA assay and the conventional culture method with Campylobacter charcoal differential agar (CCDA). A few false-negative results were obtained by both NASBA (1.42%) and CCDA (2.86%) isolation. Yet the use of enrichment culture and NASBA shortened the analysis time from 6 days to 26 h. The relative simplicity and rapidity of the NASBA assay make it an attractive alternative for detection of C. jejuni in food samples. PMID:7747955

  11. Campylobacter jejuni Fatal Sepsis in a Patient with Non-Hodgkin’s Lymphoma: Case Report and Literature Review of a Difficult Diagnosis

    PubMed Central

    Gallo, Maria Teresa; Di Domenico, Enea Gino; Toma, Luigi; Marchesi, Francesco; Pelagalli, Lorella; Manghisi, Nicola; Ascenzioni, Fiorentina; Prignano, Grazia; Mengarelli, Andrea; Ensoli, Fabrizio

    2016-01-01

    Campylobacter jejuni (C. jejuni) bacteremia is difficult to diagnose in individuals with hematological disorders undergoing chemotherapy. The cause can be attributed to the rarity of this infection, to the variable clinical presentation, and to the partial overlapping symptoms underlying the disease. Here, we report a case of a fatal sepsis caused by C. jejuni in a 76-year-old Caucasian man with non-Hodgkin’s lymphoma. After chemotherapeutic treatment, the patient experienced fever associated with severe neutropenia and thrombocytopenia without hemodynamic instability, abdominal pain, and diarrhea. The slow growth of C. jejuni in the blood culture systems and the difficulty in identifying it with conventional biochemical phenotyping methods contributed to the delay of administering a targeted antimicrobial treatment, leading to a fatal outcome. Early recognition and timely intervention are critical for the successful management of C. jejuni infection. Symptoms may be difficult to recognize in immunocompromised patients undergoing chemotherapy. Thus, it is important to increase physician awareness regarding the clinical manifestations of C. jejuni to improve therapeutic efficacy. Moreover, the use of more aggressive empirical antimicrobial treatments with aminoglycosides and/or carbapenems should be considered in immunosuppressed patients, in comparison to those currently indicated in the guidelines for cancer-related infections supporting the use of cephalosporins as monotherapy. PMID:27077849

  12. THE FREEZING POINT DEPRESSION OF MAMMALIAN TISSUES AFTER SUDDEN HEATING IN BOILING DISTILLED WATER

    PubMed Central

    Appelboom, Johannes W. Th.; Brodsky, William A.; Tuttle, William S.; Diamond, Israel

    1958-01-01

    The calculated freezing point depression of freshly excised boiled mammalian tissue is approximately the same as that of plasma. The boiling procedure was chosen to eliminate the influence of metabolism on the level of the freezing point depression. Problems created by the boiling, such as equilibrium between tissue and diluent, change in activity coefficient by dilution, and loss of CO2 content, are discussed. A frozen crushed tissue homogenate is hypertonic to plasma. Boiling and dilution of such hypertonic homogenate exposed to room temperature for 5 to 15 minutes did not produce significant or unexplicable decreases in its osmotic activity. Moreover, freezing and crushing of a boiled diluted tissue did not produce any increase of the isoosmotic level of freezing point depression. It is possible to explain these data either with the hypothesis of hypertonic cell fluid or with that of isotonic cell fluid. In the case of an assumed isotonic cell fluid, data can be explained with one assumption, experimentally backed. In the case of an assumed hypertonic theory data can be explained only with the help of at least three ad hoc postulates. The data support the validity of the classical concept which holds that cell fluid is isotonic to extracellular fluid. PMID:13563805

  13. Quantitative RNA-seq analysis of the Campylobacter jejuni transcriptome

    PubMed Central

    Chaudhuri, Roy R.; Yu, Lu; Kanji, Alpa; Perkins, Timothy T.; Gardner, Paul P.; Choudhary, Jyoti; Maskell, Duncan J.

    2011-01-01

    Campylobacter jejuni is the most common bacterial cause of foodborne disease in the developed world. Its general physiology and biochemistry, as well as the mechanisms enabling it to colonize and cause disease in various hosts, are not well understood, and new approaches are required to understand its basic biology. High-throughput sequencing technologies provide unprecedented opportunities for functional genomic research. Recent studies have shown that direct Illumina sequencing of cDNA (RNA-seq) is a useful technique for the quantitative and qualitative examination of transcriptomes. In this study we report RNA-seq analyses of the transcriptomes of C. jejuni (NCTC11168) and its rpoN mutant. This has allowed the identification of hitherto unknown transcriptional units, and further defines the regulon that is dependent on rpoN for expression. The analysis of the NCTC11168 transcriptome was supplemented by additional proteomic analysis using liquid chromatography-MS. The transcriptomic and proteomic datasets represent an important resource for the Campylobacter research community. PMID:21816880

  14. Campylobacter jejuni Colonization in Wild Birds: Results from an Infection Experiment

    PubMed Central

    Waldenström, Jonas; Axelsson-Olsson, Diana; Olsen, Björn; Hasselquist, Dennis; Griekspoor, Petra; Jansson, Lena; Teneberg, Susann; Svensson, Lovisa; Ellström, Patrik

    2010-01-01

    Campylobacter jejuni is a common cause of bacterial gastroenteritis in most parts of the world. The bacterium has a broad host range and has been isolated from many animals and environments. To investigate shedding patterns and putative effects on an avian host, we developed a colonization model in which a wild bird species, the European Robin Erithacus rubecula, was inoculated orally with C. jejuni from either a human patient or from another wild bird species, the Song Thrush Turdus philomelos. These two isolates were genetically distinct from each other and provoked very different host responses. The Song Thrush isolate colonized all challenged birds and colonization lasted 6.8 days on average. Birds infected with this isolate also showed a transient but significant decrease in body mass. The human isolate did not colonize the birds and could be detected only in the feces of the birds shortly after inoculation. European Robins infected with the wild bird isolate generated a specific antibody response to C. jejuni membrane proteins from the avian isolate, which also was cross-reactive to membrane proteins of the human isolate. In contrast, European Robins infected with the human isolate did not mount a significant response to bacterial membrane proteins from either of the two isolates. The difference in colonization ability could indicate host adaptations. PMID:20140204

  15. Wide but variable distribution of a hypervirulent Campylobacter jejuni clone in beef and dairy cattle in the united states

    USDA-ARS?s Scientific Manuscript database

    Campylobacter jejuni is a significant concern for ruminant health and food safety. Recently, a highly pathogenic C. jejuni clone (named SA) has emerged as the predominant cause of ruminant abortion and a significant cause of foodborne illnesses in the United States. Despite the recent advance in und...

  16. Updated Campylobacter jejuni capsule PCR multiplex typing system and its application to clinical isolates from south and southeast Asia

    USDA-ARS?s Scientific Manuscript database

    Campylobacter jejuni produces a polysaccharide capsule that is the major determinant of the Penner serotyping scheme. This passive slide agglutination typing system was developed in the early 1980’s and was recognized for over two decades as gold standard for C. jejuni typing. A preliminary multiple...

  17. Investigating Antibacterial Effects of Garlic (Allium sativum) Concentrate and Garlic-Derived Organosulfur Compounds on Campylobacter jejuni by Using Fourier Transform Infrared Spectroscopy, Raman Spectroscopy, and Electron Microscopy ▿ †

    PubMed Central

    Lu, Xiaonan; Rasco, Barbara A.; Jabal, Jamie M. F.; Aston, D. Eric; Lin, Mengshi; Konkel, Michael E.

    2011-01-01

    Fourier transform infrared (FT-IR) spectroscopy and Raman spectroscopy were used to study the cell injury and inactivation of Campylobacter jejuni from exposure to antioxidants from garlic. C. jejuni was treated with various concentrations of garlic concentrate and garlic-derived organosulfur compounds in growth media and saline at 4, 22, and 35°C. The antimicrobial activities of the diallyl sulfides increased with the number of sulfur atoms (diallyl sulfide < diallyl disulfide < diallyl trisulfide). FT-IR spectroscopy confirmed that organosulfur compounds are responsible for the substantial antimicrobial activity of garlic, much greater than those of garlic phenolic compounds, as indicated by changes in the spectral features of proteins, lipids, and polysaccharides in the bacterial cell membranes. Confocal Raman microscopy (532-nm-gold-particle substrate) and Raman mapping of a single bacterium confirmed the intracellular uptake of sulfur and phenolic components. Scanning electron microscopy (SEM) and transmission electron microscopy (TEM) were employed to verify cell damage. Principal-component analysis (PCA), discriminant function analysis (DFA), and soft independent modeling of class analogs (SIMCA) were performed, and results were cross validated to differentiate bacteria based upon the degree of cell injury. Partial least-squares regression (PLSR) was employed to quantify and predict actual numbers of healthy and injured bacterial cells remaining following treatment. PLSR-based loading plots were investigated to further verify the changes in the cell membrane of C. jejuni treated with organosulfur compounds. We demonstrated that bacterial injury and inactivation could be accurately investigated by complementary infrared and Raman spectroscopies using a chemical-based, “whole-organism fingerprint” with the aid of chemometrics and electron microscopy. PMID:21642409

  18. The impact of environmental conditions on Campylobacter jejuni survival in broiler faeces and litter.

    PubMed

    Smith, Shaun; Meade, Joseph; Gibbons, James; McGill, Kevina; Bolton, Declan; Whyte, Paul

    2016-01-01

    Campylobacter jejuni is the leading bacterial food-borne pathogen within the European Union, and poultry meat is an important vehicle for its transmission to humans. However, there is limited knowledge about how this organism persists in broiler litter and faeces. The aim of this study was to assess the impact of a number of environmental parameters, such as temperature, humidity, and oxygen, on Campylobacter survival in both broiler litter and faeces. Used litter was collected from a Campylobacter-negative broiler house after final depopulation and fresh faeces were collected from transport crates. Samples were confirmed as Campylobacter negative according to modified ISO methods for veterinary samples. Both sample matrices were inoculated with 9 log10 CFU/ml C. jejuni and incubated under high (≥85%) and low (≤70%) relative humidity conditions at three different temperatures (20°C, 25°C, and 30°C) under both aerobic and microaerophilic atmospheres. Inoculated litter samples were then tested for Campylobacter concentrations at time zero and every 2 hours for 12 hours, while faecal samples were examined at time zero and every 24 hours for 120 hours. A two-tailed t-test assuming unequal variance was used to compare mean Campylobacter concentrations in samples under the various temperature, humidity, and atmospheric conditions. C. jejuni survived significantly longer (P≤0.01) in faeces, with a minimum survival time of 48 hours, compared with 4 hours in used broiler litter. C. jejuni survival was significantly enhanced at 20°C in all environmental conditions in both sample matrices tested compared with survival at 25°C and 30°C. In general, survival was greater in microaerophilic compared with aerobic conditions in both sample matrices. Humidity, at the levels examined, did not appear to significantly impact C. jejuni survival in any sample matrix. The persistence of Campylobacter in broiler litter and faeces under various environmental conditions has

  19. Boiling of the interface between two immiscible liquids below the bulk boiling temperatures of both components.

    PubMed

    Pimenova, Anastasiya V; Goldobin, Denis S

    2014-11-01

    We consider the problem of boiling of the direct contact of two immiscible liquids. An intense vapour formation at such a direct contact is possible below the bulk boiling points of both components, meaning an effective decrease of the boiling temperature of the system. Although the phenomenon is known in science and widely employed in technology, the direct contact boiling process was thoroughly studied (both experimentally and theoretically) only for the case where one of liquids is becoming heated above its bulk boiling point. On the contrary, we address the case where both liquids remain below their bulk boiling points. In this paper we construct the theoretical description of the boiling process and discuss the actualisation of the case we consider for real systems.

  20. Survival of Escherichia coli, enterococci and Campylobacter jejuni in Canada goose faeces on pasture.

    PubMed

    Moriarty, E M; Weaver, L; Sinton, L W; Gilpin, B

    2012-11-01

    Freshly excreted Canada goose faeces pose a public health risk as they contain pathogenic microorganisms. Accordingly, a study was carried out on the growth and survival of resident indicator bacteria (enterococci and Escherichia coli) and inoculated Campylobacter jejuni in freshly excreted faeces over summer and winter. Canada goose faeces were collected, mixed thoroughly and inoculated with 10⁸ g⁻¹ C. jejuni. The faeces were mixed again before making the Canada goose dropping. The simulated goose droppings (N = 70) were placed on pasture, and the concentrations of E. coli, enterococci and the pathogen, C. jejuni, were monitored. In summer only, the molecular marker of E. coli LacZ and the avian-associated bacteria E2 was also monitored. Results of the survival study indicated that significant growth of enterococci and E. coli occurred in summer, before concentrations decreased to less than 15% of the original concentration (day 77) for enterococci and 0.01% for E. coli. LacZ followed a similar pattern to E. coli, while the E2 marker dropped to below 0.1% of the original concentration within 4 days. In winter, enterococci grew slightly, while no growth of E. coli occurred. In both summer and winter, C. jejuni was rapidly inactivated. This research highlights the ability of bacterial indicators to replicate and survive in the environment when harboured by avian faeces, and the limited risk aged Canada goose faeces pose as an environmental source of Campylobacter spp. © 2012 Blackwell Verlag GmbH.

  1. Characterization of Two Campylobacter jejuni Strains for Use in Volunteer Experimental-Infection Studies▿ †

    PubMed Central

    Poly, Frédéric; Read, Timothy D.; Chen, Yu-Han; Monteiro, Mario A.; Serichantalergs, Oralak; Pootong, Piyarat; Bodhidatta, Ladaporn; Mason, Carl J.; Rockabrand, David; Baqar, Shahida; Porter, Chad K.; Tribble, David; Darsley, Michael; Guerry, Patricia

    2008-01-01

    The development of vaccines against Campylobacter jejuni would be facilitated by the ability to perform phase II challenge studies. However, molecular mimicry of the lipooligosaccharide (LOS) of most C. jejuni strains with human gangliosides presents safety concerns about the development of Guillain-Barré syndrome. Clinical isolates of C. jejuni that appeared to lack genes for the synthesis of ganglioside mimics were identified by DNA probe analyses. Two clinical isolates from Southeast Asia (strains BH-01-0142 and CG8421) were determined to express the LOS type containing N-acetyl quinovosamine. No ganglioside structures were observed to be present in the LOSs of these strains, and pyrosequence analyses of the genomes of both strains confirmed the absence of genes involved in ganglioside mimicry. The capsule polysaccharide (CPS) of BH-01-0142 was determined to be composed of galactose (Gal), 6-deoxy-ido-heptose, and, in smaller amounts, d-glycero-d-ido-heptose, and the CPS of CG8421 was observed to contain Gal, 6-deoxy-altro-heptose, N-acetyl-glucosamine, and minor amounts of 6-deoxy-3-O-Me-altro-heptose. Both CPSs were shown to carry O-methyl-phosphoramidate. The two genomes contained strain-specific zones, some of which could be traced to a plasmid origin, and both contained a large chromosomal insertion related to the CJEI3 element of C. jejuni RM1221. The genomes of both strains shared a high degree of similarity to each other and, with the exception of the capsule locus of CG8421, to the type strain of the HS3 serotype, TGH9011. PMID:18809665

  2. The design of a capsule polysaccharide conjugate vaccine against Campylobacter jejuni serotype HS15.

    PubMed

    Bertolo, Lisa; Ewing, Cheryl P; Maue, Alexander; Poly, Frederic; Guerry, Patricia; Monteiro, Mario A

    2013-01-25

    Campylobacter jejuni infection is now the main cause of diarrhea-related illnesses in humans. An efficacious vaccine for the traveler and developing world market would be welcomed. We are engaged in the discovery and characterization of serotype-specific C. jejuni capsule polysaccharides (CPSs) to study their role in virulence and as protective vaccine antigens. Our prototype conjugate vaccine with serotype HS23 CPS (strain 81-176) has been shown to fully protect non-human primates against diarrhea inflicted by C. jejuni HS23, but ultimately, a useful CPS-based vaccine will have to be multivalent. To this end, we describe here the creation of a CPS-conjugate vaccine against C. jejuni serotype HS15. Structural analysis revealed that a repeating block consisting of L-α-arabinofuranose (Ara) and 6-deoxy-L-α-gulo-heptopyranose (6d-gulo-Hep) comprised the CPS of serotype HS15 type strain ATCC 43442 [→3)-α-L-Araf-(1→3)-6d-L-α-gulo-Hepp(1→](n). Strategically, the non-reducing end of the CPS was activated and used in the attachment of CPS to CRM₁₉₇ to yield a conjugate vaccine. A serological assessment of the CPS(HS15)-CRM₁₉₇ conjugate with an anti-HS15 polyclonal antibody confirmed the conservation of antigenic epitopes, and subsequent inoculation of mice with CPS(HS15)-CRM₁₉₇ revealed that this conjugate was indeed capable of raising anti-CPS(HS15) antibodies. Copyright © 2012 Elsevier Ltd. All rights reserved.

  3. Cj1386 is an ankyrin-containing protein involved in heme trafficking to catalase in Campylobacter jejuni.

    PubMed

    Flint, Annika; Sun, Yi-Qian; Stintzi, Alain

    2012-01-01

    Campylobacter jejuni, a microaerophilic bacterium, is the most frequent cause of human bacterial gastroenteritis. C. jejuni is exposed to harmful reactive oxygen species (ROS) produced during its own normal metabolic processes and during infection from the host immune system and from host intestinal microbiota. These ROS will damage DNA and proteins and cause peroxidation of lipids. Consequently, identifying ROS defense mechanisms is important for understanding how Campylobacter survives this environmental stress during infection. Construction of a ΔCj1386 isogenic deletion mutant and phenotypic assays led to its discovery as a novel oxidative stress defense gene. The ΔCj1386 mutant has an increased sensitivity toward hydrogen peroxide. The Cj1386 gene is located directly downstream from katA (catalase) in the C. jejuni genome. A ΔkatAΔ Cj1386 double deletion mutant was constructed and exhibited a sensitivity to hydrogen peroxide similar to that seen in the ΔCj1386 and ΔkatA single deletion mutants. This observation suggests that Cj1386 may be involved in the same detoxification pathway as catalase. Despite identical KatA abundances, catalase activity assays showed that the ΔCj1386 mutant had a reduced catalase activity relative to that of wild-type C. jejuni. Heme quantification of KatA protein from the ΔCj1386 mutant revealed a significant decrease in heme concentration. This indicates an important role for Cj1386 in heme trafficking to KatA within C. jejuni. Interestingly, the ΔCj1386 mutant had a reduced ability to colonize the ceca of chicks and was outcompeted by the wild-type strain for colonization of the gastrointestinal tract of neonate piglets. These results indicate an important role for Cj1386 in Campylobacter colonization and pathogenesis.

  4. Cj1386 Is an Ankyrin-Containing Protein Involved in Heme Trafficking to Catalase in Campylobacter jejuni

    PubMed Central

    Flint, Annika; Sun, Yi-Qian

    2012-01-01

    Campylobacter jejuni, a microaerophilic bacterium, is the most frequent cause of human bacterial gastroenteritis. C. jejuni is exposed to harmful reactive oxygen species (ROS) produced during its own normal metabolic processes and during infection from the host immune system and from host intestinal microbiota. These ROS will damage DNA and proteins and cause peroxidation of lipids. Consequently, identifying ROS defense mechanisms is important for understanding how Campylobacter survives this environmental stress during infection. Construction of a ΔCj1386 isogenic deletion mutant and phenotypic assays led to its discovery as a novel oxidative stress defense gene. The ΔCj1386 mutant has an increased sensitivity toward hydrogen peroxide. The Cj1386 gene is located directly downstream from katA (catalase) in the C. jejuni genome. A ΔkatAΔ Cj1386 double deletion mutant was constructed and exhibited a sensitivity to hydrogen peroxide similar to that seen in the ΔCj1386 and ΔkatA single deletion mutants. This observation suggests that Cj1386 may be involved in the same detoxification pathway as catalase. Despite identical KatA abundances, catalase activity assays showed that the ΔCj1386 mutant had a reduced catalase activity relative to that of wild-type C. jejuni. Heme quantification of KatA protein from the ΔCj1386 mutant revealed a significant decrease in heme concentration. This indicates an important role for Cj1386 in heme trafficking to KatA within C. jejuni. Interestingly, the ΔCj1386 mutant had a reduced ability to colonize the ceca of chicks and was outcompeted by the wild-type strain for colonization of the gastrointestinal tract of neonate piglets. These results indicate an important role for Cj1386 in Campylobacter colonization and pathogenesis. PMID:22081390

  5. Evaluation of different Campylobacter jejuni isolates to colonize the intestinal tract of commercial turkey poults and selective media for enumeration

    USDA-ARS?s Scientific Manuscript database

    Consumption of contaminated poultry products is the main source of human campylobacteriosis, which Campylobacter jejuni is responsible for 90 percent of human cases. Although chickens are believed to be a main source of human exposure to C. jejuni, turkey also contributes to cases of human infection...

  6. The Influence of Prior Modes of Growth, Temperature, Medium, and Substrate Surface on Biofilm Formation by Antibiotic-Resistant Campylobacter jejuni.

    PubMed

    Teh, Amy Huei Teen; Lee, Sui Mae; Dykes, Gary A

    2016-12-01

    Campylobacter jejuni is one of the most common causes of bacterial gastrointestinal food-borne infection worldwide. It has been suggested that biofilm formation may play a role in survival of these bacteria in the environment. In this study, the influence of prior modes of growth (planktonic or sessile), temperatures (37 and 42 °C), and nutrient conditions (nutrient broth and Mueller-Hinton broth) on biofilm formation by eight C. jejuni strains with different antibiotic resistance profiles was examined. The ability of these strains to form biofilm on different abiotic surfaces (stainless steel, glass, and polystyrene) as well as factors potentially associated with biofilm formation (bacterial surface hydrophobicity, auto-aggregation, and initial attachment) was also determined. The results showed that cells grown as sessile culture generally have a greater ability to form biofilm (P < 0.05) compared to their planktonic counterparts. Biofilm was also greater (P < 0.05) in lower nutrient media, while growth at different temperatures affects biofilm formation in a strain-dependent manner. The strains were able to attach and form biofilms on different abiotic surfaces, but none of them demonstrated strong, complex, or structured biofilm formation. There were no clear trends between the bacterial surface hydrophobicity, auto-aggregation, attachment, and biofilm formation by the strains. This finding suggests that environmental factors did affect biofilm formation by C. jejuni, and they are more likely to persist in the environment in the form of mixed-species rather than monospecies biofilms.

  7. The effects of high-pressure treatments on Campylobacter jejuni in ground poultry products containing polyphosphate additives.

    PubMed

    Gunther, Nereus W; Sites, Joseph; Sommers, Christopher

    2015-09-01

    Marinades containing polyphosphates have been previously implicated in the enhanced survival of Campylobacter spp. in poultry product exudates. The enhanced Campylobacter survival has been attributed primarily to the ability of some polyphosphates to change the pH of the exudate to one more amenable to Campylobacter. In this study a ground poultry product contaminated with a 6 strain Campylobacter jejuni cocktail was utilized to determine if the efficiency of high-hydrostatic-pressure treatments was negatively impacted by the presence of commonly utilized polyphosphates. Two polyphosphates, hexametaphosphate and sodium tripolyphosphate, used at 2 concentrations, 0.25 and 0.5%, failed to demonstrate any significant negative effects on the efficiency of inactivation of C. jejuni by high-pressure treatment. However, storage at 4°C of the ground poultry samples containing C. jejuni after high-pressure treatment appeared to provide a synergistic effect on Campylobacter inactivation. High-pressure treatment in conjunction with 7 d of storage at 4°C resulted in a mean reduction in C. jejuni survival that was larger than the sum of the individual reductions caused by high pressure or 4°C storage when applied separately. Published by Oxford University Press on behalf of Poultry Science Association 2015. This work is written by (a) US Government employee(s) and is in the public domain in the US.

  8. Molecular evidence for zoonotic transmission of an emergent, highly pathogenic Campylobacter jejuni clone in the United States.

    PubMed

    Sahin, Orhan; Fitzgerald, Collette; Stroika, Steven; Zhao, Shaohua; Sippy, Rachel J; Kwan, Patrick; Plummer, Paul J; Han, Jing; Yaeger, Michael J; Zhang, Qijing

    2012-03-01

    Campylobacter jejuni is a major zoonotic pathogen. A highly virulent, tetracycline-resistant C. jejuni clone (clone SA) has recently emerged in ruminant reservoirs and has become the predominant cause of sheep abortion in the United States. To determine whether clone SA is associated with human disease, we compared the clinical isolates of clone SA from sheep abortions with the human isolates of the PulseNet National Campylobacter databases at the CDC and the FDA using pulsed-field gel electrophoresis (PFGE), multilocus sequence typing (MLST), and serotyping. The combined SmaI and KpnI PFGE pattern designations of clone SA from sheep were indistinguishable from those of 123 (9.03%) human C. jejuni isolates (total, 1,361) in the CDC database, among which 56 were associated with sporadic infections and 67 were associated with outbreaks that occurred in multiple states from 2003 to 2010. Most of the outbreaks were attributed to raw milk, while the sources for most of the sporadic cases were unknown. All clone SA isolates examined, including PFGE-matched human isolates, belong to sequence type 8 (ST-8) by MLST and serotype HS:1,8, further indicating the clonality of the related isolates from different host species. Additionally, C. jejuni clone SA was identified in raw milk, cattle feces, the feces and bile of healthy sheep, and abortion cases of cattle and goats, indicating the broad distribution of this pathogenic clone in ruminants. These results provide strong molecular and epidemiological evidence for zoonotic transmission of this emergent clone from ruminants to humans and indicate that C. jejuni clone SA is an important threat to public health.

  9. Molecular Evidence for Zoonotic Transmission of an Emergent, Highly Pathogenic Campylobacter jejuni Clone in the United States

    PubMed Central

    Sahin, Orhan; Fitzgerald, Collette; Stroika, Steven; Zhao, Shaohua; Sippy, Rachel J.; Kwan, Patrick; Plummer, Paul J.; Han, Jing; Yaeger, Michael J.

    2012-01-01

    Campylobacter jejuni is a major zoonotic pathogen. A highly virulent, tetracycline-resistant C. jejuni clone (clone SA) has recently emerged in ruminant reservoirs and has become the predominant cause of sheep abortion in the United States. To determine whether clone SA is associated with human disease, we compared the clinical isolates of clone SA from sheep abortions with the human isolates of the PulseNet National Campylobacter databases at the CDC and the FDA using pulsed-field gel electrophoresis (PFGE), multilocus sequence typing (MLST), and serotyping. The combined SmaI and KpnI PFGE pattern designations of clone SA from sheep were indistinguishable from those of 123 (9.03%) human C. jejuni isolates (total, 1,361) in the CDC database, among which 56 were associated with sporadic infections and 67 were associated with outbreaks that occurred in multiple states from 2003 to 2010. Most of the outbreaks were attributed to raw milk, while the sources for most of the sporadic cases were unknown. All clone SA isolates examined, including PFGE-matched human isolates, belong to sequence type 8 (ST-8) by MLST and serotype HS:1,8, further indicating the clonality of the related isolates from different host species. Additionally, C. jejuni clone SA was identified in raw milk, cattle feces, the feces and bile of healthy sheep, and abortion cases of cattle and goats, indicating the broad distribution of this pathogenic clone in ruminants. These results provide strong molecular and epidemiological evidence for zoonotic transmission of this emergent clone from ruminants to humans and indicate that C. jejuni clone SA is an important threat to public health. PMID:22189122

  10. Sensitivity of Direct Culture, Enrichment and PCR for Detection of Campylobacter jejuni and C. coli in Broiler Flocks at Slaughter.

    PubMed

    Rodgers, J D; Simpkin, E; Lee, R; Clifton-Hadley, F A; Vidal, A B

    2017-06-01

    Broiler chicken flocks are a significant source of Campylobacter jejuni and Campylobacter coli that result in the major public health problem of campylobacteriosis. Accurate estimates of the prevalence of both C. coli and C. jejuni in flocks would enhance epidemiological understanding, risk assessment and control options. This study combined results from a panel of 10 detection tests (direct culture, enrichment and PCR) on caecal samples from flocks at slaughter. A parallel interpretation approach was used to determine the presence of Campylobacter spp. and for C. jejuni and C. coli individually. The sample was considered positive if at least one method detected the target and this interpretation was taken to represent a 'proxy gold standard' for detection in the absence of a gold standard reference test. The sensitivity of each individual method to detect Campylobacter spp., C. jejuni and C. coli was then estimated relative to the proxy gold standard. Enrichment in adapted Exeter broth (deficient in polymyxin B) with a resuscitation step was 100% sensitive, whilst direct culture on modified charcoal cefoperazone deoxycholate agar (mCCDA) was highly sensitive (97.9%). Enrichment methods using Preston broth and Bolton broth were significantly less sensitive. Enrichment in Exeter broth promoted the recovery of C. jejuni, whilst enrichment in Bolton broth favoured C. coli. A RT-PCR detection test could identify 80% of flocks that were co-colonised with both species. This study found that 76.3% (n = 127) of flocks were colonised with Campylobacter spp. The majority (95.9%) of Campylobacter-positive flocks were colonised with C. jejuni; however, approximately one-third of positive flocks were simultaneously colonised with both C. jejuni and C. coli. The findings highlight the impact of different detection methodologies on the accuracy of the estimated incidence of both C. jejuni and C. coli entering the abattoir within broiler flocks and the associated

  11. The complete genome sequence and annotation of a Campylobacter jejuni strain, MTVDSCj20, isolated from a naturally colonized farm-raised chicken

    USDA-ARS?s Scientific Manuscript database

    Campylobacter jejuni is a major cause of human foodborne illness worldwide with contaminated poultry products serving as a main source of human infection. C. jejuni strain MTVDSCj20 was isolated from the cecal contents of a farm-raised chicken naturally colonized with Campylobacter. The complete,...

  12. Wide but Variable Distribution of a Hypervirulent Campylobacter jejuni Clone in Beef and Dairy Cattle in the United States

    PubMed Central

    Tang, Yizhi; Meinersmann, Richard J.; Sahin, Orhan; Wu, Zuowei; Dai, Lei; Carlson, James; Plumblee Lawrence, Jodie; Genzlinger, Linda; LeJeune, Jeffrey T.

    2017-01-01

    ABSTRACT Campylobacter jejuni clone SA is the major cause of sheep abortion and contributes significantly to foodborne illnesses in the United States. Clone SA is hypervirulent because of its distinct ability to produce systemic infection and its predominant role in clinical sheep abortion. Despite the importance of clone SA, little is known about its distribution and epidemiological features in cattle. Here we describe a prospective study on C. jejuni clone SA prevalence in 35 feedlots in 5 different states in the United States and a retrospective analysis of clone SA in C. jejuni isolates collected by National Animal Health Monitoring System (NAHMS) dairy studies in 2002, 2007, and 2014. In feedlot cattle feces, the overall prevalence of Campylobacter organisms was 72.2%, 82.1% of which were C. jejuni. Clone SA accounted for 5.8% of the total C. jejuni isolates, but its prevalence varied by feedlot and state. Interestingly, starlings on the feedlots harbored C. jejuni in feces, including clone SA, suggesting that these birds may play a role in the transmission of Campylobacter. In dairy cattle, the overall prevalence of clone SA was 7.2%, but a significant decrease in the prevalence was observed from 2002 to 2014. Whole-genome sequence analysis of the dairy clone SA isolates revealed that it was genetically stable over the years and most of the isolates carried the tetracycline resistance gene tet(O) in the chromosome. These findings indicate that clone SA is widely distributed in both beef and dairy cattle and provide new insights into the molecular epidemiology of clone SA in ruminants. IMPORTANCE C. jejuni clone SA is a major cause of small-ruminant abortion and an emerging threat to food safety because of its association with foodborne outbreaks. Cattle appear to serve as a major reservoir for this pathogenic organism, but there is a major gap in our knowledge about the epidemiology of clone SA in beef and dairy cattle. By taking advantage of surveillance

  13. Wide but variable distribution of a hypervirulent Campylobacter jejuni clone in beef and dairy cattle in the United States.

    PubMed

    Tang, Yizhi; Meinersmann, Richard J; Sahin, Orhan; Wu, Zuowei; Dai, Lei; Carlson, James; Plumblee, Jodie; Genzlinger, Linda; LeJeune, Jeffrey T; Zhang, Qijing

    2017-09-29

    Campylobacter jejuni clone SA is the major cause of sheep abortion and contributes significantly to foodborne illnesses in the United States. Clone SA is hypervirulent because of its distinct ability to produce systemic infection and its predominant role in clinical sheep abortion. Despite the importance of clone SA, little is known about its distribution and epidemiological features in cattle. Here, we describe a prospective study on C. jejuni clone SA prevalence in 35 feedlots in 5 different states in the U.S. and a retrospective analysis of clone SA in C. jejuni isolates collected by National Animal Health Monitoring System (NAHMS) Dairy Studies 2002, 2007 and 2014. In feedlot cattle feces, the overall prevalence of Campylobacter was 72.2%, 82.1% of which were C. jejuni Clone SA accounted for 5.8% of the total C. jejuni isolates, but its prevalence varied by feedlot and state. Interestingly, starlings on the feedlots harbored C. jejuni in feces including clone SA, suggesting it may play a role in the transmission of Campylobacter In dairy cattle, the overall prevalence of clone SA was 7.2%, but a significant decrease in the prevalence was observed from 2002 to 2014. Whole genome sequence analysis of the dairy clone SA isolates revealed that it was genetically stable over the years and most of the isolates carried the tetracycline resistance gene tet(O ) in the chromosome. These findings indicate clone SA is widely distributed in both beef and dairy cattle, and provide new insights into the molecular epidemiology of clone SA in ruminants. Importance C. jejuni clone SA is a major cause of small ruminant abortion and an emerging threat to food safety because of its association with foodborne outbreaks. Cattle appears to serve as a major reservoir for this pathogenic organism, but there is a major gap in our knowledge about the epidemiology of clone SA in beef and dairy cattle. By taking advantage of surveillance studies conducted on a national scale, this

  14. Detection of gyrA mutation among clinical isolates of Campylobacter jejuni isolated in Egypt by MAMA-PCR.

    PubMed

    Said, Mayar M; El-Mohamady, Hanan; El-Beih, Fawkia M; Rockabrand, David M; Ismail, Tharwat F; Monteville, Marshall R; Ahmed, Salwa F; Klena, John D; Salama, Mohamed S

    2010-10-04

    Campylobacter spp are the major cause of enteritis in humans and more than 90% of reported infections are caused by Campylobacter jejuni. Fluoroquinolones such as ciprofloxacin are the antibiotics of choice for treatment. An increase in the frequency of ciprofloxacin-resistant Campylobacter has been reported globally due to a single base mutation (C-257 to T) in codon 86 of the quinolone resistance determining region (QRDR) of the gyrA gene altering the amino acid sequence from threonine at position 86 to isoleucine (Thr-86 to Ile). Campylobacter spp (n = 118) were selected from a collection of Egyptian isolates spanning 1998 to 2005. The presence of C. jejuni gyrA gene was confirmed in each isolate by a PCR assay amplifying 368 bp portion of the gyrA gene. C to T alteration was detected by the mismatch amplification mutation assay MAMA PCR. The MIC of nalidixic acid (NA) and ciprofloxacin (CIP) was determined by E-test. C. jejuni gyrA gene was detected in 100 of the Campylobacter spp studied; the other 18 isolates were found to be Campylobacter coli by lpxA PCR. The mutation was detected in 89 C. jejuni resistant isolates with MIC values (NA; 8 - >256 μg/ml) and (CIP; 4 - >32 μg/ml). The other 11 sensitive C. jejuni isolates with MIC values (NA; 0.38 - 3 µg/ml) and (CIP; 0.03 - 0.125 µg/ml) were not amplified by the MAMA primers. There was 100% congruence with MAMA PCR, MIC results and gyrA gene sequence analysis. In Egypt the main mechanism for resistance to fluoroquinolones is an alteration in the gyrA QRDR. MAMA PCR provides an economical and rapid means for screening fluoroquinolone resistance.

  15. Epidemiology, Relative Invasive Ability, Molecular Characterization, and Competitive Performance of Campylobacter jejuni Strains in the Chicken Gut▿

    PubMed Central

    Pope, Christopher; Wilson, Janet; Taboada, Eduardo N.; MacKinnon, Joanne; Felipe Alves, Cristiano A.; Nash, John H. E.; Rahn, Kris; Tannock, Gerald W.

    2007-01-01

    One hundred forty-one Campylobacter jejuni isolates from humans with diarrhea and 100 isolates from retailed poultry meat were differentiated by flaA typing. The bacteria were isolated in a specific geographical area (Dunedin) in New Zealand over a common time period. Twenty nine flaA types were detected, one of which (flaA restriction fragment length polymorphism type 15 [flaA-15]) predominated among isolates from humans (∼30% of isolates). This strain was of low prevalence (5% of isolates) among poultry isolates. flaA-15 strains were five to six times more invasive of HEp2 cells in an in vitro assay than a flaA type (flaA-3) that was commonly encountered on poultry meat (23% of isolates) but was seldom associated with human illness (5%). Competitive-exclusion experiments with chickens, utilizing real-time quantitative PCR to measure the population sizes of specific strains representing flaA-15 (T1016) and flaA-3 (Pstau) in digesta, were carried out. These experiments showed that T1016 always outcompeted Pstau in the chicken intestine. Genomic comparisons of T1016 and Pstau were made using DNA microarrays representing the genome of C. jejuni NCTC 11168. These comparisons revealed differences between the strains in the gene content of the Cj1417c-to-Cj1442c region of the genome, which is associated with the formation of capsular polysaccharide. The strains differed in Penner type (T1016, O42; Pstau, O53). It was concluded that poultry meat was at least one source of human infection with C. jejuni, that some Campylobacter strains detected in poultry meat are of higher virulence for humans than others, and that bacterial attributes affecting strain virulence and commensal colonization ability may be linked. PMID:17921281

  16. Development of Multiple-Locus Variable-Number Tandem-Repeat Analysis for Molecular Subtyping of Campylobacter jejuni by Using Capillary Electrophoresis

    PubMed Central

    Techaruvichit, Punnida; Vesaratchavest, Mongkol; Keeratipibul, Suwimon; Kuda, Takashi; Kimura, Bon

    2015-01-01

    Campylobacter jejuni is a common cause of the frequently reported food-borne diseases in developed and developing nations. This study describes the development of multiple-locus variable-number tandem-repeat (VNTR) analysis (MLVA) using capillary electrophoresis as a novel typing method for microbial source tracking and epidemiological investigation of C. jejuni. Among 36 tandem repeat loci detected by the Tandem Repeat Finder program, 7 VNTR loci were selected and used for characterizing 60 isolates recovered from chicken meat samples from retail shops, samples from chicken meat processing factory, and stool samples. The discrimination ability of MLVA was compared with that of multilocus sequence typing (MLST). MLVA (diversity index of 0.97 with 31 MLVA types) provided slightly higher discrimination than MLST (diversity index of 0.95 with 25 MLST types). The overall concordance between MLVA and MLST was estimated at 63% by adjusted Rand coefficient. MLVA predicted MLST type better than MLST predicted MLVA type, as reflected by Wallace coefficient (Wallace coefficient for MLVA to MLST versus MLST to MLVA, 86% versus 51%). MLVA is a useful tool and can be used for effective monitoring of C. jejuni and investigation of epidemics caused by C. jejuni. PMID:26025899

  17. Size-exclusion chromatography for the determination of the boiling point distribution of high-boiling petroleum fractions.

    PubMed

    Boczkaj, Grzegorz; Przyjazny, Andrzej; Kamiński, Marian

    2015-03-01

    The paper describes a new procedure for the determination of boiling point distribution of high-boiling petroleum fractions using size-exclusion chromatography with refractive index detection. Thus far, the determination of boiling range distribution by chromatography has been accomplished using simulated distillation with gas chromatography with flame ionization detection. This study revealed that in spite of substantial differences in the separation mechanism and the detection mode, the size-exclusion chromatography technique yields similar results for the determination of boiling point distribution compared with simulated distillation and novel empty column gas chromatography. The developed procedure using size-exclusion chromatography has a substantial applicability, especially for the determination of exact final boiling point values for high-boiling mixtures, for which a standard high-temperature simulated distillation would have to be used. In this case, the precision of final boiling point determination is low due to the high final temperatures of the gas chromatograph oven and an insufficient thermal stability of both the gas chromatography stationary phase and the sample. Additionally, the use of high-performance liquid chromatography detectors more sensitive than refractive index detection allows a lower detection limit for high-molar-mass aromatic compounds, and thus increases the sensitivity of final boiling point determination. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  18. Analysis of the activity and regulon of the two-component regulatory system encoded by Cjj1484 and Cjj1483 of Campylobacter jejuni

    USDA-ARS?s Scientific Manuscript database

    Campylobacter jejuni is a leading cause of bacterial diarrheal disease throughout the world and a frequent commensal in the intestinal tract of poultry and many other animals. For maintaining optimal growth and ability to colonize various hosts, C. jejuni depends upon two-component regulatory system...

  19. Campylobacter jejuni colonization and population structure in urban populations of ducks and starlings in New Zealand

    PubMed Central

    Mohan, Vathsala; Stevenson, Mark; Marshall, Jonathan; Fearnhead, Paul; Holland, Barbara R; Hotter, Grant; French, Nigel P

    2013-01-01

    Abstract A repeated cross-sectional study was conducted to determine the prevalence of Campylobacter spp. and the population structure of C. jejuni in European starlings and ducks cohabiting multiple public access sites in an urban area of New Zealand. The country's geographical isolation and relatively recent history of introduction of wild bird species, including the European starling and mallard duck, create an ideal setting to explore the impact of geographical separation on the population biology of C. jejuni, as well as potential public health implications. A total of 716 starling and 720 duck fecal samples were collected and screened for C. jejuni over a 12 month period. This study combined molecular genotyping, population genetics and epidemiological modeling and revealed: (i) higher Campylobacter spp. isolation in starlings (46%) compared with ducks (30%), but similar isolation of C. jejuni in ducks (23%) and starlings (21%), (ii) significant associations between the isolation of Campylobacter spp. and host species, sampling location and time of year using logistic regression, (iii) evidence of population differentiation, as indicated by FST, and host-genotype association with clonal complexes CC ST-177 and CC ST-682 associated with starlings, and clonal complexes CC ST-1034, CC ST-692, and CC ST-1332 associated with ducks, and (iv) greater genetic diversity and genotype richness in ducks compared with starlings. These findings provide evidence that host-associated genotypes, such as the starling-associated ST-177 and ST-682, represent lineages that were introduced with the host species in the 19th century. The isolation of sequence types associated with human disease in New Zealand indicate that wild ducks and starlings need to be considered as a potential public health risk, particularly in urban areas. We applied molecular epidemiology and population genetics to obtain insights in to the population structure, host-species relationships, gene flow and

  20. Multi-omics approaches to deciphering a hypervirulent strain of Campylobacter jejuni

    USDA-ARS?s Scientific Manuscript database

    Background: Campylobacter jejuni clone SA recently emerged as the predominant cause of sheep abortion in the U.S. and is also associated with foodborne gastroenteritis in humans. A distinct phenotype of this clone is its ability to induce bacteremia and abortion. To facilitate understanding the path...

  1. Subcooled forced convection boiling of trichlorotrifluoroethane

    NASA Technical Reports Server (NTRS)

    Dougall, R. S.; Panian, D. J.

    1972-01-01

    Experimental heat-transfer data were obtained for the forced-convection boiling of trichlorotrifluoroethane (R-113 or Freon-113) in a vertical annular test annular test section. The 97 data points obtained covered heat transfer by forced convection, local boiling, and fully-developed boiling. Correlating methods were obtained which accurately predicted the heat flux as a function of wall superheat (boiling curve) over the range of parameters studied.

  2. Optimizing the Combination of Smoking and Boiling on Quality of Korean Traditional Boiled Loin (M. longissimus dorsi)

    PubMed Central

    Choi, Yun-Sang; Kim, Hyun-Wook; Kim, Young-Boong; Kim, Cheon-Jei

    2015-01-01

    The combined effects of smoking and boiling on the proximate composition, technological quality traits, shear force, and sensory characteristics of the Korean traditional boiled loin were studied. Cooking loss, processing loss, and shear force were lower in the smoked/boiled samples than those in the control (without smoking treatment) (p<0.05). The results showed that the boiled loin samples between the control and treatment did not differ significantly in protein, fat, or ash contents, or pH values (p>0.05). The treated samples had higher score for overall acceptability than the control (p<0.05). Thus, these results show that the Korean traditional boiled loin treated with smoking for 60 min before boiling had improved physicochemical properties and sensory characteristics. PMID:26761822

  3. The myth of the boiling point.

    PubMed

    Chang, Hasok

    2008-01-01

    Around 1800, many reputable scientists reported significant variations in the temperature of pure water boiling under normal atmospheric pressure. The reported variations included a difference of over 1 degree C between boiling in metallic and glass vessels (Gay-Lussac), and "superheating" up to 112 degrees C on extracting dissolved air out of water (De Luc). I have confirmed most of these observations in my own experiments, many of which are described in this paper. Water boils at the "boiling point" only under very particular circumstances. Our common-sense intuition about the fixedness of the boiling point is only sustained by our limited experience.

  4. Universality of oscillating boiling in Leidenfrost transition

    NASA Astrophysics Data System (ADS)

    Tran, Tuan; Khavari, Mohammad

    2017-11-01

    The Leidenfrost transition leads a boiling system to the boiling crisis, a state in which the liquid loses contact with the heated surface due to excessive vapor generation. Here, using experiments of liquid droplets boiling on a heated surface, we report a new phenomenon, termed oscillating boiling, at the Leidenfrost transition. We show that oscillating boiling results from the competition between two effects: separation of liquid from the heated surface due to localized boiling, and rewetting. We argue theoretically that the Leidenfrost transition can be predicted based on its link with the oscillating boiling phenomenon, and verify the prediction experimentally for various liquids. This work was funded by Nanyang Technological University and A*STAR, Singapore.

  5. Binational outbreak of Guillain-Barré syndrome associated with Campylobacter jejuni infection, Mexico and USA, 2011.

    PubMed

    Jackson, B R; Zegarra, J Alomía; López-Gatell, H; Sejvar, J; Arzate, F; Waterman, S; Núñez, A Sánchez; López, B; Weiss, J; Cruz, R Quintero; Murrieta, D Y López; Luna-Gierke, R; Heiman, K; Vieira, A R; Fitzgerald, C; Kwan, P; Zárate-Bermúdez, M; Talkington, D; Hill, V R; Mahon, B

    2014-05-01

    In June 2011, a cluster of suspected cases of Guillain-Barré syndrome (GBS), which can follow Campylobacter jejuni infection, was identified in San Luis Río Colorado (SLRC), Sonora, Mexico and Yuma County, Arizona, USA. An outbreak investigation identified 26 patients (18 from Sonora, eight from Arizona) with onset of GBS 4 May-21 July 2011, exceeding the expected number of cases (n = 1-2). Twenty-one (81%) patients reported antecedent diarrhoea, and 61% of 18 patients tested were seropositive for C. jejuni IgM antibodies. In a case-control study matched on age group, sex, ethnicity, and neighbourhood of residence, all Arizona GBS patients travelled to SLRC during the exposure period vs. 45% of matched controls (matched odds ratio 8·1, 95% confidence interval 1·5-∞). Exposure information and an environmental assessment suggested that GBS cases resulted from a large outbreak of C. jejuni infection from inadequately disinfected tap water in SLRC. Binational collaboration was essential in investigating this cross-border GBS outbreak, the first in mainland North America since 1976.

  6. Orthogonal typing methods identify genetic diversity among Belgian Campylobacter jejuni strains isolated over a decade from poultry and cases of sporadic human illness

    USDA-ARS?s Scientific Manuscript database

    Campylobacter jejuni is a zoonotic pathogen commonly associated with human gastroenteritis. Retail poultry meat is a major food-related transmission source of C. jejuni to humans. The present study investigated the genetic diversity, clonal relationship, and strain risk-ranking of 403 representativ...

  7. Genome-Wide Identification of Host-Segregating Epidemiological Markers for Source Attribution in Campylobacter jejuni

    PubMed Central

    Thépault, Amandine; Méric, Guillaume; Rivoal, Katell; Pascoe, Ben; Mageiros, Leonardos; Touzain, Fabrice; Rose, Valérie; Béven, Véronique; Chemaly, Marianne

    2017-01-01

    ABSTRACT Campylobacter is among the most common worldwide causes of bacterial gastroenteritis. This organism is part of the commensal microbiota of numerous host species, including livestock, and these animals constitute potential sources of human infection. Molecular typing approaches, especially multilocus sequence typing (MLST), have been used to attribute the source of human campylobacteriosis by quantifying the relative abundance of alleles at seven MLST loci among isolates from animal reservoirs and human infection, implicating chicken as a major infection source. The increasing availability of bacterial genomes provides data on allelic variation at loci across the genome, providing the potential to improve the discriminatory power of data for source attribution. Here we present a source attribution approach based on the identification of novel epidemiological markers among a reference pan-genome list of 1,810 genes identified by gene-by-gene comparison of 884 genomes of Campylobacter jejuni isolates from animal reservoirs, the environment, and clinical cases. Fifteen loci involved in metabolic activities, protein modification, signal transduction, and stress response or coding for hypothetical proteins were selected as host-segregating markers and used to attribute the source of 42 French and 281 United Kingdom clinical C. jejuni isolates. Consistent with previous studies of British campylobacteriosis, analyses performed using STRUCTURE software attributed 56.8% of British clinical cases to chicken, emphasizing the importance of this host reservoir as an infection source in the United Kingdom. However, among French clinical isolates, approximately equal proportions of isolates were attributed to chicken and ruminant reservoirs, suggesting possible differences in the relative importance of animal host reservoirs and indicating a benefit for further national-scale attribution modeling to account for differences in production, behavior, and food consumption

  8. [Outbreak of gastroenteritis caused by Campylobacter jejuni transmitted through drinking water].

    PubMed

    Godoy, Pere; Artigues, Antoni; Nuín, Carmen; Aramburu, Jesús; Pérez, Montse; Domínguez, Angela; Salleras, Lluís

    2002-11-23

    The aim of this study was to conduct a clinical-epidemiological and microbiological investigation into an outbreak of waterborne disease caused by Campylobacter jejuni due to the consumption of drinking water. A historical cohort study was carried out among 237 residents of Torres de Segre (Lleida, Spain) who were selected using a systematic sample. We conducted a telephone interview about water consumption, symptoms and the onset of disease. We investigated samples of drinking water and stools from 14 patients. The risk associated with each water source was assessed by applying relative risk (RR) analysis at 95% confidence (CI) intervals. The overall attack rate was 18.3% (43/237). The symptoms were: diarrhoea, 93.0% (18/43); abdominal pain, 80.9% (34/42); nausea; 56,1% (23/41); vomits, 42.9% (18/42), and fever, 11.9% (5/42). Only 5.8% of patients contact with his physician. The consumption of drinking water was statistically associated with the disease (RR = 3.0; 95% CI, 1.7-5.3), while the consumption of bottled water (RR = 0.6; 95% CI 0.3-1.0) and water from other villages (RR = 0.3; 95% CI, 0.1-1.1) were a protection factor. The day of outbreak notification we did not detect any residual chlorine in the drinking water: it was qualified as no potable and we isolated Campylobacter jejuni in 8 samples stools. This research highlights the potential importance of waterborne outbreaks of gastroenteritis due to Campylobacter jejuni transmitted through untreated drinking water and suggests to need systematic controls over drinking water and the proper register of their results.

  9. Prevalence, antimicrobial resistance and genetic diversity of Campylobacter coli and Campylobacter jejuni in Ecuadorian broilers at slaughter age

    PubMed Central

    Vinueza-Burgos, Christian; Wautier, Magali; Martiny, Delphine; Cisneros, Marco; Van Damme, Inge; De Zutter, Lieven

    2017-01-01

    Abstract Thermotolerant Campylobacter spp. are a major cause of foodborne gastrointestinal infections worldwide. The linkage of human campylobacteriosis and poultry has been widely described. In this study we aimed to investigate the prevalence, antimicrobial resistance and genetic diversity of C. coli and C. jejuni in broilers from Ecuador. Caecal content from 379 randomly selected broiler batches originating from 115 farms were collected from 6 slaughterhouses located in the province of Pichincha during 1 year. Microbiological isolation was performed by direct plating on mCCDA agar. Identification of Campylobacter species was done by PCR. Minimum inhibitory concentration (MIC) values for gentamicin, ciprofloxacin, nalidixic acid, tetracycline, streptomycin, and erythromycin were obtained. Genetic variation was assessed by RFLP-flaA typing and Multilocus Sequence Typing (MLST) of selected isolates. Prevalence at batch level was 64.1%. Of the positive batches 68.7% were positive for C. coli, 18.9% for C. jejuni, and 12.4% for C. coli and C. jejuni. Resistance rates above 67% were shown for tetracycline, ciprofloxacin, and nalidixic acid. The resistance pattern tetracycline, ciprofloxin, and nalidixic acid was the dominant one in both Campylobacter species. RFLP-flaA typing analysis showed that C. coli and C. jejuni strains belonged to 38 and 26 profiles respectively. On the other hand MLST typing revealed that C. coli except one strain belonged to CC-828, while C. jejuni except 2 strains belonged to 12 assigned clonal complexes (CCs). Furthermore 4 new sequence types (STs) for both species were described, whereby 2 new STs for C. coli were based on new allele sequences. Further research is necessary to estimate the impact of the slaughter of Campylobacter positive broiler batches on the contamination level of carcasses in slaughterhouses and at retail in Ecuador. PMID:28339716

  10. Characterisation by multilocus sequence and porA and flaA typing of Campylobacter jejuni isolated from samples of dog faeces collected in one city in New Zealand.

    PubMed

    Mohan, V; Stevenson, M A; Marshall, J C; French, N P

    2017-07-01

    To investigate the prevalence of Campylobacter spp. and C. jejuni in dog faecal material collected from dog walkways in the city of Palmerston North, New Zealand, and to characterise the C. jejuni isolates by multilocus sequence typing (MLST) and porA and flaA antigen gene typing. A total of 355 fresh samples of dogs faeces were collected from bins provided for the disposal of dog faeces in 10 walkways in Palmerston North, New Zealand, between August 2008-July 2009. Presumptive Campylobacter colonies, cultured on modified charcoal cefoperazone deoxycholate plates, were screened for genus Campylobacter and C. jejuni by PCR. The C. jejuni isolates were subsequently characterised by MLST and porA and flaA typing, and C. jejuni sequence types (ST) were assigned. Of the 355 samples collected, 72 (20 (95% CI=16-25)%) were positive for Campylobacter spp. and 22 (6 (95% CI=4-9)%) were positive for C. jejuni. Of the 22 C. jejuni isolates, 19 were fully typed by MLST. Ten isolates were assigned to the clonal complex ST-45 and three to ST-52. The allelic combinations of ST-45/flaA 21/porA 44 (n=3), ST-45/flaA 22/porA 53 (n=3) and ST-52/ flaA 57/porA 905 (n=3) were most frequent. The successful isolation of C. jejuni from canine faecal samples collected from faecal bins provides evidence that Campylobacter spp. may survive outside the host for at least several hours despite requiring fastidious growth conditions in culture. The results show that dogs carry C. jejuni genotypes (ST-45, ST-50, ST-52 and ST-696) that have been reported in human clinical cases. Although these results do not provide any evidence either for the direction of infection or for dogs being a potential risk factor for human campylobacteriosis, dog owners are advised to practice good hygiene with respect to their pets to reduce potential exposure to infection.

  11. Odd-Boiled Eggs

    ERIC Educational Resources Information Center

    Kaminsky, Kenneth; Scheman, Naomi

    2010-01-01

    At a Shabbat lunch in Madrid not long ago, the conversation turned to the question of boiling eggs. One of the guests mentioned that a Dutch rabbi he knew had heard that in order to make it more likely that boiled eggs be kosher, you should add an egg to the pot if the number you began with was even. According to the laws of Kashruth, Jews may not…

  12. 21 CFR 872.6710 - Boiling water sterilizer.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Boiling water sterilizer. 872.6710 Section 872...) MEDICAL DEVICES DENTAL DEVICES Miscellaneous Devices § 872.6710 Boiling water sterilizer. (a) Identification. A boiling water sterilizer is an AC-powered device that consists of a container for boiling water...

  13. 21 CFR 872.6710 - Boiling water sterilizer.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Boiling water sterilizer. 872.6710 Section 872...) MEDICAL DEVICES DENTAL DEVICES Miscellaneous Devices § 872.6710 Boiling water sterilizer. (a) Identification. A boiling water sterilizer is an AC-powered device that consists of a container for boiling water...

  14. 21 CFR 872.6710 - Boiling water sterilizer.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 8 2013-04-01 2013-04-01 false Boiling water sterilizer. 872.6710 Section 872...) MEDICAL DEVICES DENTAL DEVICES Miscellaneous Devices § 872.6710 Boiling water sterilizer. (a) Identification. A boiling water sterilizer is an AC-powered device that consists of a container for boiling water...

  15. 21 CFR 872.6710 - Boiling water sterilizer.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 8 2012-04-01 2012-04-01 false Boiling water sterilizer. 872.6710 Section 872...) MEDICAL DEVICES DENTAL DEVICES Miscellaneous Devices § 872.6710 Boiling water sterilizer. (a) Identification. A boiling water sterilizer is an AC-powered device that consists of a container for boiling water...

  16. Draft Genome Sequences of Three Multiantibiotic-Resistant Campylobacter jejuni Strains (2865, 2868, and 2871) Isolated from Poultry at Retail Outlets in Malaysia

    PubMed Central

    Teh, Amy Huei Teen; Lee, Sui Mae

    2016-01-01

    Campylobacter jejuni is a frequent cause of human bacterial gastrointestinal foodborne disease worldwide. Antibiotic resistance in this species is of public health concern. The draft genome sequences of three multiantibiotic-resistant C. jejuni strains (2865, 2868, and 2871) isolated from poultry at retail outlets in Malaysia are presented here. PMID:27151799

  17. Growth phase-dependent activation of the DccRS regulon of Campylobacter jejuni

    USDA-ARS?s Scientific Manuscript database

    Two-component systems are widespread prokaryotic signal transduction devices which allow the regulation of cellular functions in response to changing environmental conditions. The two-component system DccRS (Cj1223-Cj1222) of Campylobacter jejuni is important for the colonization of chickens. Here w...

  18. Boils

    MedlinePlus

    ... boil is an infection that affects groups of hair follicles and nearby skin tissue. Related conditions include: Carbunculosis ... found on the skin's surface. Damage to the hair follicle allows the infection to grow deeper into the ...

  19. Interaction effects between sender and receiver processes in indirect transmission of Campylobacter jejuni between broilers.

    PubMed

    van Bunnik, Bram A D; Hagenaars, Thomas J; Bolder, Nico M; Nodelijk, Gonnie; de Jong, Mart C M

    2012-07-25

    Infectious diseases in plants, animals and humans are often transmitted indirectly between hosts (or between groups of hosts), i.e. via some route through the environment instead of via direct contacts between these hosts. Here we study indirect transmission experimentally, using transmission of Campylobacter jejuni (C. jejuni) between spatially separated broilers as a model system. We distinguish three stages in the process of indirect transmission; (1) an infectious "sender" excretes the agent, after which (2) the agent is transported via some route to a susceptible "receiver", and subsequently (3) the receiver becomes colonised by the agent. The role of the sender and receiver side (stage 1 and stage 3) was studied here by using acidification of the drinking water as a modulation mechanism. In the experiment one control group and three treatment groups were monitored for the presence of C. jejuni by taking daily cloacal swabs. The three treatments consisted of acidification of the drinking water of the inoculated animals (the senders), acidification of the drinking water of the susceptible animals (the receivers) or acidification of the drinking water of both inoculated and susceptible animals. In the control group 12 animals got colonised out of a possible 40, in each treatment groups 3 animals out of a possible 40 were found colonised with C. jejuni. The results of the experiments show a significant decrease in transmission rate (β) between the control groups and treatment groups (p < 0.01 for all groups) but not between different treatments; there is a significant negative interaction effect when both the sender and the receiver group receive acidified drinking water (p = 0.01). This negative interaction effect could be due to selection of bacteria already at the sender side thereby diminishing the effect of acidification at the receiver side.

  20. Interaction effects between sender and receiver processes in indirect transmission of Campylobacter jejuni between broilers

    PubMed Central

    2012-01-01

    Background Infectious diseases in plants, animals and humans are often transmitted indirectly between hosts (or between groups of hosts), i.e. via some route through the environment instead of via direct contacts between these hosts. Here we study indirect transmission experimentally, using transmission of Campylobacter jejuni (C. jejuni) between spatially separated broilers as a model system. We distinguish three stages in the process of indirect transmission; (1) an infectious “sender” excretes the agent, after which (2) the agent is transported via some route to a susceptible “receiver”, and subsequently (3) the receiver becomes colonised by the agent. The role of the sender and receiver side (stage 1 and stage 3) was studied here by using acidification of the drinking water as a modulation mechanism. Results In the experiment one control group and three treatment groups were monitored for the presence of C. jejuni by taking daily cloacal swabs. The three treatments consisted of acidification of the drinking water of the inoculated animals (the senders), acidification of the drinking water of the susceptible animals (the receivers) or acidification of the drinking water of both inoculated and susceptible animals. In the control group 12 animals got colonised out of a possible 40, in each treatment groups 3 animals out of a possible 40 were found colonised with C. jejuni. Conclusions The results of the experiments show a significant decrease in transmission rate (β) between the control groups and treatment groups (p < 0.01 for all groups) but not between different treatments; there is a significant negative interaction effect when both the sender and the receiver group receive acidified drinking water (p = 0.01). This negative interaction effect could be due to selection of bacteria already at the sender side thereby diminishing the effect of acidification at the receiver side. PMID:22831274

  1. Pool Boiling Experiment Has Five Successful Flights

    NASA Technical Reports Server (NTRS)

    Chiaramonte, Fran

    1997-01-01

    The Pool Boiling Experiment (PBE) is designed to improve understanding of the fundamental mechanisms that constitute nucleate pool boiling. Nucleate pool boiling is a process wherein a stagnant pool of liquid is in contact with a surface that can supply heat to the liquid. If the liquid absorbs enough heat, a vapor bubble can be formed. This process occurs when a pot of water boils. On Earth, gravity tends to remove the vapor bubble from the heating surface because it is dominated by buoyant convection. In the orbiting space shuttle, however, buoyant convection has much less of an effect because the forces of gravity are very small. The Pool Boiling Experiment was initiated to provide insight into this nucleate boiling process, which has many earthbound applications in steamgeneration power plants, petroleum plants, and other chemical plants. In addition, by using the test fluid R-113, the Pool Boiling Experiment can provide some basic understanding of the boiling behavior of cryogenic fluids without the large cost of an experiment using an actual cryogen.

  2. Gene expression profiling of chicken cecal tonsils and ileum following oral exposure to soluble and PLGA-encapsulated CpG ODN, and lysate of Campylobacter jejuni.

    PubMed

    Taha-Abdelaziz, Khaled; Alkie, Tamiru Negash; Hodgins, Douglas C; Yitbarek, Alexander; Shojadoost, Bahram; Sharif, Shayan

    2017-12-01

    Campylobacter jejuni (C. jejuni) is a leading bacterial cause of food-borne illness in humans. Contaminated chicken meat is an important source of infection for humans. Chickens are not clinically affected by colonization, and immune responses following natural infection have limited effects on bacterial load in the gut. Induction of intestinal immune responses may possibly lead to a breakdown of the commensal relationship of chickens with Campylobacter. We have recently shown that soluble and poly D, L-lactic-co-glycolic acid (PLGA)-encapsulated CpG oligodeoxynucleotide (ODN) as well as C. jejuni lysate, are effective in reducing the intestinal burden of C. jejuni in chickens; however, the mechanisms behind this protection have yet to be determined. The present study was undertaken to investigate the mechanisms of host responses conferred by these treatments. Chickens were treated orally with soluble CpG ODN, or PLGA-encapsulated CpG ODN, or C. jejuni lysate, and expression of cytokines and antimicrobial peptides was evaluated in cecal tonsils and ileum using quantitative RT-PCR. Oral administration of soluble CpG ODN upregulated the expression of interferon (IFN)-γ, interleukin (IL)-1β, CXCLi2, transforming growth factor (TGF)-β4/1, IL-10 and IL-13, while treatment with PLGA-encapsulated CpG ODN upregulated the expression of IL-1β, CXCLi2, TGF-β4/1, IL-13, avian β-defensin (AvBD) 1, AvBD2 and cathelicidin 3 (CATHL-3). C. jejuni lysate upregulated the expression of IFN-γ, IL-1β, TGF-β4/1, IL-13, AvBD1, and CATHL-3. In conclusion, induction of cytokine and antimicrobial peptides expression in intestinal microenvironments may provide a means of reducing C. jejuni colonization in broiler chickens, a key step in reducing the incidence of campylobacteriosis in humans. Copyright © 2017 Elsevier B.V. All rights reserved.

  3. The influence of dissolved oxygen level and medium on biofilm formation by Campylobacter jejuni.

    PubMed

    Teh, Amy Huei Teen; Lee, Sui Mae; Dykes, Gary A

    2017-02-01

    Campylobacter jejuni survival in aerobic environments has been suggested to be mediated by biofilm formation. Biofilm formation by eight C. jejuni strains under both aerobic and microaerobic conditions in different broths (Mueller-Hinton (MH), Bolton and Brucella) was quantified. The dissolved oxygen (DO) content of the broths under both incubation atmospheres was determined. Biofilm formation for all strains was highest in MH broth under both incubation atmospheres. Four strains had lower biofilm formation in MH under aerobic as compared to microaerobic incubation, while biofilm formation by the other four strains did not differ under the 2 atm. Two strains had higher biofilm formation under aerobic as compared to microaerobic atmospheres in Bolton broth. Biofilm formation by all other strains in Bolton, and all strains in Brucella broth, did not differ under the 2 atm. Under aerobic incubation DO levels in MH > Brucella > Bolton broth. Under microaerobic conditions levels in MH = Brucella > Bolton broth. Levels of DO in MH and Brucella broth were lower under microaerobic conditions but those of Bolton did not differ under the 2 atm. Experimental conditions and especially the DO of broth media confound previous conclusions drawn about aerobic biofilm formation by C. jejuni. Copyright © 2016 Elsevier Ltd. All rights reserved.

  4. Molecular epidemiology of Campylobacter jejuni infection in Israel-a nationwide study.

    PubMed

    Weinberger, M; Moran-Gilad, J; Rokney, A; Davidov, Y; Agmon, V; Peretz, C; Valinsky, L

    2016-12-01

    The incidence of Campylobacter infection in Israel, particularly among children <2 years of age, has risen over the last decade and became one of the highest among industrialized countries. This study explored the molecular epidemiology of Campylobacter jejuni in Israel over a decade (2003-2012) using multilocus sequence typing (MLST) combined with demographic metadata. Representative clinical isolates (438) from a large national repository together with selected veterinary isolates (74) were subject to MLST. The distribution of age groups, ethnicity and clinical source across various genotypes was evaluated using Poisson modelling. The 512 studied isolates were assigned 126 distinct sequence types (STs) (18.8% novel STs) grouped into 21 clonal complexes (CCs). Most human, poultry and bovine STs clustered together in the leading CCs. Three dominant STs (ST21, ST6608, ST4766) were detected only since 2006. Patients infected with the leading CCs were similarly distributed along densely populated areas. The frequency of blood isolates was higher in patients infected with CC353 (relative rate (RR)=2.0, 95% CI 1.03-3.9, adjusted p value (adj.p) 0.047) and CC42 (RR=4.4, 95% CI 1.7-11.6, adj.p 0.018) and lower with CC257 (RR=0.3, 95% CI 0.1-0.9, adj. p 0.047). The distribution of age groups and ethnicity also varied across the leading CCs. In conclusion, C. jejuni isolates in a national sample appeared highly diverse with a high proportion of new STs. Phylogenic analysis was compatible with poultry and cattle as possible food sources of clinical infection. Demographic characteristics of the infected patients coupled with strain invasiveness across different genotypes revealed a complex epidemiology of C. jejuni transmission in Israel. Copyright © 2016 European Society of Clinical Microbiology and Infectious Diseases. Published by Elsevier Ltd. All rights reserved.

  5. Selection for pro-inflammatory mediators produces chickens more resistant to Campylobacter jejuni

    USDA-ARS?s Scientific Manuscript database

    Campylobacter spp. are the second leading cause of bacterial-induced foodborne illnesses with an estimated economic burden of nearly $2 billion per year. Most human illness associated with campylobacteriosis is due to infection by C. jejuni and chickens are recognized as a reservoir, which could le...

  6. The complete genome sequences of 65 Campylobacter jejuni and C. coli strains

    USDA-ARS?s Scientific Manuscript database

    Campylobacter jejuni (Cj) and C. coli (Cc) are genetically highly diverse based on various molecular methods including MLST, microarray-based comparisons and the whole genome sequences of a few strains. Cj and Cc diversity is also exhibited by variable capsular polysaccharides (CPS) that are the maj...

  7. Analysis of evolutionary patterns of genes in campylobacter jejuni and C. coli

    USDA-ARS?s Scientific Manuscript database

    Background: In order to investigate the population genetics structure of thermophilic Campylobacter spp., we extracted a set of 1029 core gene families (CGF) from 25 sequenced genomes of C. jejuni, C. coli and C. lari. Based on these CGFs we employed different approaches to reveal the evolutionary ...

  8. Genetic diversity and clonal characteristics of ciprofloxacin-resistant Campylobacter jejuni isolated from Chilean patients with gastroenteritis.

    PubMed

    Collado, Luis; Muñoz, Nataly; Porte, Lorena; Ochoa, Sofía; Varela, Carmen; Muñoz, Ivo

    2018-03-01

    Campylobacter jejuni is a major cause of acute gastroenteritis worldwide. However, it has also been associated with other diseases such as bacteremia and with several post-infection sequelae. Although campylobacteriosis is usually a self-limited infection, antibiotics are indicated for severe and chronic conditions. Unfortunately, several industrialised nations have reported a substantial increase in antibiotic resistance of C. jejuni. However, there is still a lack of knowledge about the epidemiology of resistance developed by this pathogen in the developing world. For this reason, our objective was to determine the resistance of clinical C. jejuni strains to ciprofloxacin and erythromycin in Chile and their associated genotypes. Fifty C. jejuni isolates recovered from fecal samples of people with acute gastroenteritis, in central and southern Chile between 2006 and 2015, were analysed. Resistance to erythromycin and ciprofloxacin was assessed by disk diffusion and agar dilution methods. Furthermore, these strains were genotyped by Multilocus Sequence Typing (MLST). Only one of the isolates was resistant to erythromycin. However, 48% of them were resistant to ciprofloxacin. The minimal inhibitory concentration of these ciprofloxacin-resistant isolates was in the range between 4 and 32 μg/ml. Moreover, MLST analyses showed that most ciprofloxacin-resistant strains were grouped into three dominant clonal complexes (ST-21, ST-48 and ST-353), while the unique strain resistant to both antibiotics belonged to the ST-45 complex. Our results evidence a high ciprofloxacin resistance and suggest that there is a dissemination of resistant clonal lineages responsible for cases of campylobacteriosis in Chile. Further studies should elucidate the origin of these resistant genotypes. Copyright © 2018 Elsevier B.V. All rights reserved.

  9. Bactericidal effect of hydrolysable and condensed tannin extracts on Campylobacter jejuni in vitro

    USDA-ARS?s Scientific Manuscript database

    Strategies are sought to reduce intestinal colonization of food-producing animals by Campylobacter jejuni, a leading bacterial cause of human foodborne illness worldwide. Presently, we tested the antimicrobial activity of hydrolysable-rich blackberry, cranberry, chestnut tannin extracts, and conden...

  10. Development of a loop-mediated isothermal amplification method for rapid campylobacter jejuni detection

    USDA-ARS?s Scientific Manuscript database

    Introduction: Campylobacter jejuni is the leading foodborne pathogen that causes human bacterial gastroenteritis worldwide. Poultry products are regarded as a major source for human infection. Early, rapid detection of this microorganism in poultry products is necessary for contamination control ...

  11. Guillain Barré Syndrome is induced in Non-Obese Diabetic (NOD) mice following Campylobacter jejuni infection and is exacerbated by antibiotics.

    PubMed

    St Charles, J L; Bell, J A; Gadsden, B J; Malik, A; Cooke, H; Van de Grift, L K; Kim, H Y; Smith, E J; Mansfield, L S

    2017-02-01

    Campylobacter jejuni is a leading cause of bacterial gastroenteritis linked to several serious autoimmune sequelae such as the peripheral neuropathies Guillain Barré syndrome (GBS) and Miller Fisher syndrome (MFS). We hypothesized that GBS and MFS can result in NOD wild type (WT) mice or their congenic interleukin (IL)-10 or B7-2 knockouts secondary to C. jejuni infection. Mice were gavaged orally with C. jejuni strains HB93-13 and 260.94 from patients with GBS or CF93-6 from a patient with MFS and assessed for clinical neurological signs and phenotypes, anti-ganglioside antibodies, and cellular infiltrates and lesions in gut and peripheral nerve tissues. Significant increases in autoantibodies against single gangliosides (GM1, GQ1b, GD1a) occurred in infected NOD mice of all genotypes, although the isotypes varied (NOD WT had IgG1, IgG3; NOD B7-2 -/- had IgG3; NOD IL-10 -/- had IgG1, IgG3, IgG2a). Infected NOD WT and NOD IL-10 -/- mice also produced anti-ganglioside antibodies of the IgG1 isotype directed against a mixture of GM1/GQ1b gangliosides. Phenotypic tests showed significant differences between treatment groups of all mouse genotypes. Peripheral nerve lesions with macrophage infiltrates were significantly increased in infected mice of NOD WT and IL-10 -/- genotypes compared to sham-inoculated controls, while lesions with T cell infiltrates were significantly increased in infected mice of the NOD B7-2 -/- genotype compared to sham-inoculated controls. In both infected and sham inoculated NOD IL-10 -/- mice, antibiotic treatment exacerbated neurological signs, lesions and the amount and number of different isotypes of antiganglioside autoantibodies produced. Thus, inducible mouse models of post-C. jejuni GBS are feasible and can be characterized based on evaluation of three factors-onset of GBS clinical signs/phenotypes, anti-ganglioside autoantibodies and nerve lesions. Based on these factors we characterized 1) NOD B-7 -/- mice as an acute

  12. Draft Genome Sequences of Three Multiantibiotic-Resistant Campylobacter jejuni Strains (2865, 2868, and 2871) Isolated from Poultry at Retail Outlets in Malaysia.

    PubMed

    Teh, Amy Huei Teen; Lee, Sui Mae; Dykes, Gary A

    2016-05-05

    Campylobacter jejuni is a frequent cause of human bacterial gastrointestinal foodborne disease worldwide. Antibiotic resistance in this species is of public health concern. The draft genome sequences of three multiantibiotic-resistant C. jejuni strains (2865, 2868, and 2871) isolated from poultry at retail outlets in Malaysia are presented here. Copyright © 2016 Teh et al.

  13. Efficacy of Peracetic acid and Zinc in reducing Campylobacter jejuni on chicken skin

    USDA-ARS?s Scientific Manuscript database

    Campylobacter jejuni is a leading cause of bacterial foodborne disease in humans worldwide, largely associated with the consumption of contaminated poultry products. With increasing consumer demand for natural and minimally processed foods, the use of Generally Recognized as Safe status antimicrobia...

  14. Cryogenic Boil-Off Reduction System

    NASA Astrophysics Data System (ADS)

    Plachta, David W.; Guzik, Monica C.

    2014-03-01

    A computational model of the cryogenic boil-off reduction system being developed by NASA as part of the Cryogenic Propellant Storage and Transfer technology maturation project has been applied to a range of propellant storage tanks sizes for high-performing in-space cryogenic propulsion applications. This effort focuses on the scaling of multi-layer insulation (MLI), cryocoolers, broad area cooling shields, radiators, solar arrays, and tanks for liquid hydrogen propellant storage tanks ranging from 2 to 10 m in diameter. Component scaling equations were incorporated into the Cryogenic Analysis Tool, a spreadsheet-based tool used to perform system-level parametric studies. The primary addition to the evolution of this updated tool is the integration of a scaling method for reverse turbo-Brayton cycle cryocoolers, as well as the development and inclusion of Self-Supporting Multi-Layer Insulation. Mass, power, and sizing relationships are traded parametrically to establish the appropriate loiter period beyond which this boil-off reduction system application reduces mass. The projected benefit compares passive thermal control to active thermal control, where active thermal control is evaluated for reduced boil-off with a 90 K shield, zero boil-off with a single heat interception stage at the tank wall, and zero boil-off with a second interception stage at a 90 K shield. Parametric studies show a benefit over passive storage at loiter durations under one month, in addition to showing a benefit for two-stage zero boil-off in terms of reducing power and mass as compared to single stage zero boil-off. Furthermore, active cooling reduces the effect of varied multi-layer insulation performance, which, historically, has been shown to be significant.

  15. Correlational approach to turbulent saturated film boiling

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Chu, T.Y.

    A correlation method for saturated film boiling is proposed. The correlation is based on the analogy between film boiling and natural convection. As in the case of natural convection, the turbulent film boiling correlation takes the form of a Nusselt number versus the Raleigh number power law, Nu[sub B] [proportional to] Ra[sub B][sup 1.3]. The proposed correlation shows very good agreement with current data for film boiling of water from vertical surfaces. The general applicability of the correlation is established by comparisons with film boiling data from R-113 and cryogenic fluids. 25 refs., 8 figs.

  16. Evaporation, Boiling and Bubbles

    ERIC Educational Resources Information Center

    Goodwin, Alan

    2012-01-01

    Evaporation and boiling are both terms applied to the change of a liquid to the vapour/gaseous state. This article argues that it is the formation of bubbles of vapour within the liquid that most clearly differentiates boiling from evaporation although only a minority of chemistry textbooks seems to mention bubble formation in this context. The…

  17. Film boiling of mercury droplets

    NASA Technical Reports Server (NTRS)

    Baumeister, K. J.; Schoessow, G. J.; Chmielewski, C. E.

    1975-01-01

    Vaporization times of mercury droplets in Leidenfrost film boiling on a flat horizontal plate are measured in an air atmosphere. Extreme care was used to prevent large amplitude droplet vibrations and surface wetting; therefore, these data can be compared to film boiling theory. Diffusion from the upper surface of the drop appears as a dominant mode of mass transfer from the drop. A closed-form analytical film boiling theory is developed to account for the diffusive evaporation. Reasonable agreement between data and theory is seen.

  18. Genetic Basis and Clonal Population Structure of Antibiotic Resistance in Campylobacter jejuni Isolated From Broiler Carcasses in Belgium.

    PubMed

    Elhadidy, Mohamed; Miller, William G; Arguello, Hector; Álvarez-Ordóñez, Avelino; Duarte, Alexandra; Dierick, Katelijne; Botteldoorn, Nadine

    2018-01-01

    Human campylobacteriosis is the leading food-borne zoonosis in industrialized countries. This study characterized the clonal population structure, antimicrobial resistance profiles and occurrence of antimicrobial resistance determinants of a set of Campylobacter jejuni strains isolated from broiler carcasses in Belgium. Minimum inhibitory concentrations (MICs) against five commonly-used antibiotics (ciprofloxacin, nalidixic acid, tetracycline, gentamicin, and erythromycin) were determined for 204 C. jejuni isolates. More than half of the isolates were resistant to ciprofloxacin or nalidixic acid. In contrast, a lower percentage of screened isolates were resistant to gentamicin or erythromycin. C. jejuni isolates resistant to ciprofloxacin and/or nalidixic acid were screened for the substitution T86I in the quinolone resistance determining region (QRDR) of the gyrA gene, while C. jejuni isolates resistant to tetracycline were screened for the presence of the tet(O) gene. These resistance determinants were observed in most but not all resistant isolates. Regarding resistance to erythromycin, different mutations occurred in diverse genetic loci, including mutations in the 23S rRNA gene, the rplD and rplV ribosomal genes, and the intergenic region between cmeR and cmeABC . Interestingly, and contrary to previous reports, the A2075G transition mutation in the 23S rRNA gene was only found in one strain displaying a high level of resistance to erythromycin. Ultimately, molecular typing by multilocus sequence typing revealed that two sequence types (ST-824 and ST-2274) were associated to quinolones resistance by the presence of mutations in the gene gyrA ( p = 0.01). In addition, ST-2274 was linked to the CIP-NAL-TET-AMR multidrug resistant phenotype. In contrast, clonal complex CC-45 was linked to increased susceptibility to the tested antibiotics. The results obtained in this study provide better understanding of the phenotypic and the molecular basis of antibiotic

  19. Differential Distribution of Type II CRISPR-Cas Systems in Agricultural and Nonagricultural Campylobacter coli and Campylobacter jejuni Isolates Correlates with Lack of Shared Environments

    PubMed Central

    Pearson, Bruce M.; Louwen, Rogier; van Baarlen, Peter; van Vliet, Arnoud H.M.

    2015-01-01

    CRISPR (clustered regularly interspaced palindromic repeats)-Cas (CRISPR-associated) systems are sequence-specific adaptive defenses against phages and plasmids which are widespread in prokaryotes. Here we have studied whether phylogenetic relatedness or sharing of environmental niches affects the distribution and dissemination of Type II CRISPR-Cas systems, first in 132 bacterial genomes from 15 phylogenetic classes, ranging from Proteobacteria to Actinobacteria. There was clustering of distinct Type II CRISPR-Cas systems in phylogenetically distinct genera with varying G+C%, which share environmental niches. The distribution of CRISPR-Cas within a genus was studied using a large collection of genome sequences of the closely related Campylobacter species Campylobacter jejuni (N = 3,746) and Campylobacter coli (N = 486). The Cas gene cas9 and CRISPR-repeat are almost universally present in C. jejuni genomes (98.0% positive) but relatively rare in C. coli genomes (9.6% positive). Campylobacter jejuni and agricultural C. coli isolates share the C. jejuni CRISPR-Cas system, which is closely related to, but distinct from the C. coli CRISPR-Cas system found in C. coli isolates from nonagricultural sources. Analysis of the genomic position of CRISPR-Cas insertion suggests that the C. jejuni-type CRISPR-Cas has been transferred to agricultural C. coli. Conversely, the absence of the C. coli-type CRISPR-Cas in agricultural C. coli isolates may be due to these isolates not sharing the same environmental niche, and may be affected by farm hygiene and biosecurity practices in the agricultural sector. Finally, many CRISPR spacer alleles were linked with specific multilocus sequence types, suggesting that these can assist molecular epidemiology applications for C. jejuni and C. coli. PMID:26338188

  20. Enabling Highly Effective Boiling from Superhydrophobic Surfaces

    NASA Astrophysics Data System (ADS)

    Allred, Taylor P.; Weibel, Justin A.; Garimella, Suresh V.

    2018-04-01

    A variety of industrial applications such as power generation, water distillation, and high-density cooling rely on heat transfer processes involving boiling. Enhancements to the boiling process can improve the energy efficiency and performance across multiple industries. Highly wetting textured surfaces have shown promise in boiling applications since capillary wicking increases the maximum heat flux that can be dissipated. Conversely, highly nonwetting textured (superhydrophobic) surfaces have been largely dismissed for these applications as they have been shown to promote formation of an insulating vapor film that greatly diminishes heat transfer efficiency. The current Letter shows that boiling from a superhydrophobic surface in an initial Wenzel state, in which the surface texture is infiltrated with liquid, results in remarkably low surface superheat with nucleate boiling sustained up to a critical heat flux typical of hydrophilic wetting surfaces, and thus upends this conventional wisdom. Two distinct boiling behaviors are demonstrated on both micro- and nanostructured superhydrophobic surfaces based on the initial wetting state. For an initial surface condition in which vapor occupies the interstices of the surface texture (Cassie-Baxter state), premature film boiling occurs, as has been commonly observed in the literature. However, if the surface texture is infiltrated with liquid (Wenzel state) prior to boiling, drastically improved thermal performance is observed; in this wetting state, the three-phase contact line is pinned during vapor bubble growth, which prevents the development of a vapor film over the surface and maintains efficient nucleate boiling behavior.

  1. Campylobacter jejuni colonization and population structure in urban populations of ducks and starlings in New Zealand.

    PubMed

    Mohan, Vathsala; Stevenson, Mark; Marshall, Jonathan; Fearnhead, Paul; Holland, Barbara R; Hotter, Grant; French, Nigel P

    2013-08-01

    A repeated cross-sectional study was conducted to determine the prevalence of Campylobacter spp. and the population structure of C. jejuni in European starlings and ducks cohabiting multiple public access sites in an urban area of New Zealand. The country's geographical isolation and relatively recent history of introduction of wild bird species, including the European starling and mallard duck, create an ideal setting to explore the impact of geographical separation on the population biology of C. jejuni, as well as potential public health implications. A total of 716 starling and 720 duck fecal samples were collected and screened for C. jejuni over a 12 month period. This study combined molecular genotyping, population genetics and epidemiological modeling and revealed: (i) higher Campylobacter spp. isolation in starlings (46%) compared with ducks (30%), but similar isolation of C. jejuni in ducks (23%) and starlings (21%), (ii) significant associations between the isolation of Campylobacter spp. and host species, sampling location and time of year using logistic regression, (iii) evidence of population differentiation, as indicated by FST , and host-genotype association with clonal complexes CC ST-177 and CC ST-682 associated with starlings, and clonal complexes CC ST-1034, CC ST-692, and CC ST-1332 associated with ducks, and (iv) greater genetic diversity and genotype richness in ducks compared with starlings. These findings provide evidence that host-associated genotypes, such as the starling-associated ST-177 and ST-682, represent lineages that were introduced with the host species in the 19th century. The isolation of sequence types associated with human disease in New Zealand indicate that wild ducks and starlings need to be considered as a potential public health risk, particularly in urban areas. © 2013 The Authors. Microbiology Open published by John Wiley & Sons Ltd.

  2. Matrix metalloproteinases-2 and -9 in Campylobacter jejuni-induced paralytic neuropathy resembling Guillain-Barré syndrome in chickens.

    PubMed

    Nyati, Kishan Kumar; Prasad, Kashi Nath; Agrawal, Vinita; Husain, Nuzhat

    2017-10-01

    Inflammation in Guillain-Barré syndrome (GBS) is manifested by changes in matrix metalloproteinase (MMP) and pro-inflammatory cytokine expression. We investigated the expression of MMP-2, -9 and TNF-α and correlated it with pathological changes in sciatic nerve tissue from Campylobacter jejuni-induced chicken model for GBS. Campylobacter jejuni and placebo were fed to chickens and assessed for disease symptoms. Sciatic nerves were examined by histopathology and immunohistochemistry. Expressions of MMPs and TNF-α, were determined by real-time PCR, and activities of MMPs by zymography. Diarrhea developed in 73.3% chickens after infection and 60.0% of them developed GBS like neuropathy. Pathology in sciatic nerves showed perinodal and/or patchy demyelination, perivascular focal lymphocytic infiltration and myelin swelling on 10th- 20th post infection day (PID). MMP-2, -9 and TNF-α were up-regulated in progressive phase of the disease. Enhanced MMP-2, -9 and TNF-α production in progressive phase correlated with sciatic nerve pathology in C. jejuni-induced GBS chicken model. Copyright © 2017 Elsevier Ltd. All rights reserved.

  3. Cluster Analysis of Campylobacter jejuni Genotypes Isolated from Small and Medium-Sized Mammalian Wildlife and Bovine Livestock from Ontario Farms.

    PubMed

    Viswanathan, M; Pearl, D L; Taboada, E N; Parmley, E J; Mutschall, S K; Jardine, C M

    2017-05-01

    Using data collected from a cross-sectional study of 25 farms (eight beef, eight swine and nine dairy) in 2010, we assessed clustering of molecular subtypes of C. jejuni based on a Campylobacter-specific 40 gene comparative genomic fingerprinting assay (CGF40) subtypes, using unweighted pair-group method with arithmetic mean (UPGMA) analysis, and multiple correspondence analysis. Exact logistic regression was used to determine which genes differentiate wildlife and livestock subtypes in our study population. A total of 33 bovine livestock (17 beef and 16 dairy), 26 wildlife (20 raccoon (Procyon lotor), five skunk (Mephitis mephitis) and one mouse (Peromyscus spp.) C. jejuni isolates were subtyped using CGF40. Dendrogram analysis, based on UPGMA, showed distinct branches separating bovine livestock and mammalian wildlife isolates. Furthermore, two-dimensional multiple correspondence analysis was highly concordant with dendrogram analysis showing clear differentiation between livestock and wildlife CGF40 subtypes. Based on multilevel logistic regression models with a random intercept for farm of origin, we found that isolates in general, and raccoons more specifically, were significantly more likely to be part of the wildlife branch. Exact logistic regression conducted gene by gene revealed 15 genes that were predictive of whether an isolate was of wildlife or bovine livestock isolate origin. Both multiple correspondence analysis and exact logistic regression revealed that in most cases, the presence of a particular gene (13 of 15) was associated with an isolate being of livestock rather than wildlife origin. In conclusion, the evidence gained from dendrogram analysis, multiple correspondence analysis and exact logistic regression indicates that mammalian wildlife carry CGF40 subtypes of C. jejuni distinct from those carried by bovine livestock. Future studies focused on source attribution of C. jejuni in human infections will help determine whether wildlife

  4. Key Role of Capsular Polysaccharide in the Induction of Systemic Infection and Abortion by Hypervirulent Campylobacter jejuni

    PubMed Central

    Sahin, Orhan; Terhorst, Samantha A.; Burrough, Eric R.; Shen, Zhangqi; Wu, Zuowei; Dai, Lei; Tang, Yizhi; Plummer, Paul J.; Ji, Ju; Yaeger, Michael J.

    2017-01-01

    ABSTRACT Campylobacter jejuni is a zoonotic pathogen, and a hypervirulent clone, named clone SA, has recently emerged as the predominant cause of ovine abortion in the United States. To induce abortion, orally ingested Campylobacter must translocate across the intestinal epithelium, spread systemically in the circulation, and reach the fetoplacental tissue. Bacterial factors involved in these steps are not well understood. C. jejuni is known to produce capsular polysaccharide (CPS), but the specific role that CPS plays in systemic infection and particularly abortion in animals remains to be determined. In this study, we evaluated the role of CPS in bacteremia using a mouse model and in abortion using a pregnant guinea pig model following oral challenge. Compared with C. jejuni NCTC 11168 and 81-176, a clone SA isolate (IA3902) resulted in significantly higher bacterial counts and a significantly longer duration of bacteremia in mice. The loss of capsule production via gene-specific mutagenesis in IA3902 led to the complete abolishment of bacteremia in mice and abortion in pregnant guinea pigs, while complementation of capsule expression almost fully restored these phenotypes. The capsule mutant strain was also impaired for survival in guinea pig sera and sheep blood. Sequence-based analyses revealed that clone SA possesses a unique CPS locus with a mosaic structure, which has been stably maintained in all clone SA isolates derived from various hosts and times. These findings establish CPS as a key virulence factor for the induction of systemic infection and abortion in pregnant animals and provide a viable candidate for the development of vaccines against hypervirulent C. jejuni. PMID:28373351

  5. On the pulse boiling frequency in thermosyphons

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Liu, J.F.; Wang, J.C.Y.

    1992-02-01

    The unsteady periodic boiling phenomenon, pulse boiling, appearing in the evaporator of thermosyphons has been mentioned and investigated by many researchers. The heat transfer coefficient in evaporators was predicted according to different considerations of flow patterns. For instance, Shiraishi et al. proposed a method based on a combination flow pattern: the nucleate boiling in a liquid pool and the evaporation from a falling condensate film. Liu et al. only considered a pure pulse boiling flow pattern, and Xin et al. focused on the flow pattern of the continuous boiling process without pulse phenomenon. Besides, the forming conditions of pulse boilingmore » were also described differently. Xin et al. also reported that pulse boiling cannot occur in a carbon-steel/water heat pipe; Ma et al., however, observed this phenomenon in a carbon-steel/water thermosyphon. Nearly all researchers mentioned that this phenomenon indeed exists in glass/water thermosyphons. Although the influential factors have been discussed qualitatively, the quantitative analysis has yet to be conducted. This study focuses on the pulse boiling frequency as a criterion for the determination of flow patterns, and attempts are made to predict the frequency both experimentally and theoretically.« less

  6. Differential Distribution of Type II CRISPR-Cas Systems in Agricultural and Nonagricultural Campylobacter coli and Campylobacter jejuni Isolates Correlates with Lack of Shared Environments.

    PubMed

    Pearson, Bruce M; Louwen, Rogier; van Baarlen, Peter; van Vliet, Arnoud H M

    2015-09-02

    CRISPR (clustered regularly interspaced palindromic repeats)-Cas (CRISPR-associated) systems are sequence-specific adaptive defenses against phages and plasmids which are widespread in prokaryotes. Here we have studied whether phylogenetic relatedness or sharing of environmental niches affects the distribution and dissemination of Type II CRISPR-Cas systems, first in 132 bacterial genomes from 15 phylogenetic classes, ranging from Proteobacteria to Actinobacteria. There was clustering of distinct Type II CRISPR-Cas systems in phylogenetically distinct genera with varying G+C%, which share environmental niches. The distribution of CRISPR-Cas within a genus was studied using a large collection of genome sequences of the closely related Campylobacter species Campylobacter jejuni (N = 3,746) and Campylobacter coli (N = 486). The Cas gene cas9 and CRISPR-repeat are almost universally present in C. jejuni genomes (98.0% positive) but relatively rare in C. coli genomes (9.6% positive). Campylobacter jejuni and agricultural C. coli isolates share the C. jejuni CRISPR-Cas system, which is closely related to, but distinct from the C. coli CRISPR-Cas system found in C. coli isolates from nonagricultural sources. Analysis of the genomic position of CRISPR-Cas insertion suggests that the C. jejuni-type CRISPR-Cas has been transferred to agricultural C. coli. Conversely, the absence of the C. coli-type CRISPR-Cas in agricultural C. coli isolates may be due to these isolates not sharing the same environmental niche, and may be affected by farm hygiene and biosecurity practices in the agricultural sector. Finally, many CRISPR spacer alleles were linked with specific multilocus sequence types, suggesting that these can assist molecular epidemiology applications for C. jejuni and C. coli. © The Author(s) 2015. Published by Oxford University Press on behalf of the Society for Molecular Biology and Evolution.

  7. Genotyping of Campylobacter jejuni from broiler carcasses and slaughterhouse environment by amplified fragment length polymorphism.

    PubMed

    Johnsen, G; Kruse, H; Hofshagen, M

    2006-12-01

    We examined the occurrence and diversity of Campylobacter jejuni on broiler carcasses during slaughter of an infected flock and in the slaughterhouse environment during slaughter and postdisinfection before a new production run. During the slaughter of a known C. jejuni infected broiler flock, samples were taken from broiler carcasses at 7 different stages during the process. Thirty-seven sites in the slaughterhouse environment were sampled both during process and postdisinfection. The samples were analyzed for C. jejuni, and genetic fingerprinting was performed using amplified fragment length polymorphism. All carcass samples were positive. Of the environmental samples collected during slaughter, 89% were positive; 100% of those from the arrival, stunning, scalding, defeathering, and evisceration facilities and 67% of those from the cooling and sorting facilities. Postdisinfection, 41% of the samples were positive; 71% of those from the arrival and stunning area, 60% of those from the scalding and defeathering area, and 20% of those from the evisceration, cooling, and sorting area. The C. jejuni isolates (n = 60) recovered were grouped into 4 different amplified fragment length polymorphism clones with a similarity index of 95% or greater. All isolates obtained from the flock and 94% of the isolates obtained from the environment during slaughtering belonged to clone A, whereas 1 environmental isolate belonged to each of the clones B and C. Isolates from clones A, B, and D were present postdisinfection. Only clone B was detected on flocks slaughtered during the previous week. The high level and continuous presence of Campylobacter in the environment constitutes a risk for transmission to negative carcasses. In Norway, where above 96% of the broiler flocks are Campylobacter-negative, this aspect is of special importance. The ability of Campylobacter to remain in the slaughterhouse environment through washing and disinfection is associated with constructional

  8. Noise analysis of nucleate boiling

    NASA Technical Reports Server (NTRS)

    Mcknight, R. D.; Ram, K. S.

    1971-01-01

    The techniques of noise analysis have been utilized to investigate nucleate pool boiling. A simple experimental setup has been developed for obtaining the power spectrum of a nucleate boiling system. These techniques were first used to study single bubbles, and a method of relating the two-dimensional projected size and the local velocity of the bubbles to the auto-correlation functions is presented. This method is much less time consuming than conventional methods of measurement and has no probes to disturb the system. These techniques can be used to determine the contribution of evaporation to total heat flux in nucleate boiling. Also, these techniques can be used to investigate the effect of various parameters upon the frequency response of nucleate boiling. The predominant frequencies of the power spectrum correspond to the frequencies of bubble generation. The effects of heat input, degree of subcooling, and liquid surface tension upon the power spectra of a boiling system are presented. It was found that the degree of subcooling has a more pronounced effect upon bubble size than does heat flux. Also the effect of lowering surface tension can be sufficient to reduce the effect of the degree of subcooling upon the size of the bubbles.

  9. A direct-sensing galactose chemoreceptor recently evolved in invasive strains of Campylobacter jejuni

    NASA Astrophysics Data System (ADS)

    Day, Christopher J.; King, Rebecca M.; Shewell, Lucy K.; Tram, Greg; Najnin, Tahria; Hartley-Tassell, Lauren E.; Wilson, Jennifer C.; Fleetwood, Aaron D.; Zhulin, Igor B.; Korolik, Victoria

    2016-10-01

    A rare chemotaxis receptor, Tlp11, has been previously identified in invasive strains of Campylobacter jejuni, the most prevalent cause of bacterial gastroenteritis worldwide. Here we use glycan and small-molecule arrays, as well as surface plasmon resonance, to show that Tlp11 specifically interacts with galactose. Tlp11 is required for the chemotactic response of C. jejuni to galactose, as shown using wild type, allelic inactivation and addition mutants. The inactivated mutant displays reduced virulence in vivo, in a model of chicken colonization. The Tlp11 sensory domain represents the first known sugar-binding dCache_1 domain, which is the most abundant family of extracellular sensors in bacteria. The Tlp11 signalling domain interacts with the chemotaxis scaffolding proteins CheV and CheW, and comparative genomic analysis indicates a likely recent evolutionary origin for Tlp11. We propose to rename Tlp11 as CcrG, Campylobacter ChemoReceptor for Galactose.

  10. Multilocus Sequence Types of Campylobacter jejuni Isolates from Different Sources in Eastern China.

    PubMed

    Zhang, Gong; Zhang, Xiaoyan; Hu, Yuanqing; Jiao, Xin-An; Huang, Jinlin

    2015-09-01

    Campylobacter jejuni is a major food-borne pathogen that causes human gastroenteritis in many developed countries. In our study, we applied multilocus sequence typing (MLST) technology to 167 C. jejuni isolates from diverse sources in Eastern China to examine their genetic diversity. MLST defined 94 sequence types (STs) belonging to 18 clonal complexes (CCs). Forty-five STs from 60 isolates (36%) and 22 alleles have not been previously documented in an international database. One hundred and two isolates, accounting for 61.1% of all isolates, belonged to eight clonal complexes. The eight major CCs were also the most common complexes from different sources. The most common ST type of isolates from human and food was ST-353. The dominant ST type in chicken and foods was ST-354. Among 21 STs that contained two or more different sources isolates, 15 STs contained human isolates and isolates from other sources, suggesting that potentially pathogenic strains are not restricted to specific lineages.

  11. A direct-sensing galactose chemoreceptor recently evolved in invasive strains of Campylobacter jejuni

    DOE PAGES

    Day, Christopher J.; King, Rebecca M.; Shewell, Lucy K.; ...

    2016-10-20

    A rare chemotaxis receptor, Tlp11, has been previously identified in invasive strains of Campylobacter jejuni, the most prevalent cause of bacterial gastroenteritis worldwide. Here we use glycan and small-molecule arrays, as well as surface plasmon resonance, to show that Tlp11 specifically interacts with galactose. Tlp11 is required for the chemotactic response of C. jejuni to galactose, as shown using wild type, allelic inactivation and addition mutants. The inactivated mutant displays reduced virulence in vivo, in a model of chicken colonization. The Tlp11 sensory domain represents the first known sugar-binding dCache_1 domain, which is the most abundant family of extracellular sensorsmore » in bacteria. The Tlp11 signalling domain interacts with the chemotaxis scaffolding proteins CheV and CheW, and comparative genomic analysis indicates a likely recent evolutionary origin for Tlp11. Lastly, we propose to rename Tlp11 as CcrG, Campylobacter ChemoReceptor for Galactose.« less

  12. A direct-sensing galactose chemoreceptor recently evolved in invasive strains of Campylobacter jejuni

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Day, Christopher J.; King, Rebecca M.; Shewell, Lucy K.

    A rare chemotaxis receptor, Tlp11, has been previously identified in invasive strains of Campylobacter jejuni, the most prevalent cause of bacterial gastroenteritis worldwide. Here we use glycan and small-molecule arrays, as well as surface plasmon resonance, to show that Tlp11 specifically interacts with galactose. Tlp11 is required for the chemotactic response of C. jejuni to galactose, as shown using wild type, allelic inactivation and addition mutants. The inactivated mutant displays reduced virulence in vivo, in a model of chicken colonization. The Tlp11 sensory domain represents the first known sugar-binding dCache_1 domain, which is the most abundant family of extracellular sensorsmore » in bacteria. The Tlp11 signalling domain interacts with the chemotaxis scaffolding proteins CheV and CheW, and comparative genomic analysis indicates a likely recent evolutionary origin for Tlp11. Lastly, we propose to rename Tlp11 as CcrG, Campylobacter ChemoReceptor for Galactose.« less

  13. Effect of oxygen stress on growth and survival of Clostridium perfringens, Campylobacter jejuni, and Listeria monocytogenes under different storage conditions.

    PubMed

    Al-Qadiri, Hamzah; Sablani, Shyam S; Ovissipour, Mahmoudreza; Al-Alami, Nivin; Govindan, Byju; Rasco, Barbara

    2015-04-01

    This study investigated the growth and survival of three foodborne pathogens (Clostridium perfringens, Campylobacter jejuni, and Listeria monocytogenes) in beef (7% fat) and nutrient broth under different oxygen levels. Samples were tested under anoxic (<0.5%), microoxic (6 to 8%), and oxic (20%) conditions during storage at 7 °C for 14 days and at 22 °C for 5 days. Two initial inoculum concentrations were used (1 and 2 log CFU per g of beef or per ml of broth). The results show that C. perfringens could grow in beef at 22 °C, with an increase of approximately 5 log under anoxic conditions and a 1-log increase under microoxic conditions. However, C. perfringens could not survive in beef held at 7 °C under microoxic and oxic storage conditions after 14 days. In an anoxic environment, C. perfringens survived in beef samples held at 7 °C, with a 1-log reduction. A cell decline was observed at 2 log under these conditions, with no surviving cells at the 1-log level. However, the results show that C. jejuni under microoxic conditions survived with declining cell numbers. Significant increases in L. monocytogenes (5 to 7 log) were observed in beef held at 22 °C for 5 days, with the lowest levels recovered under anoxic conditions. L. monocytogenes in refrigerated storage increased by a factor of 2 to 4 log. It showed the greatest growth under oxic conditions, with significant growth under anoxic conditions. These findings can be used to enhance food safety in vacuum-packed and modified atmosphere-packaged food products.

  14. Evaluation of Two Multiplex PCR-High-Resolution Melt Curve Analysis Methods for Differentiation of Campylobacter jejuni and Campylobacter coli Intraspecies.

    PubMed

    Banowary, Banya; Dang, Van Tuan; Sarker, Subir; Connolly, Joanne H; Chenu, Jeremy; Groves, Peter; Raidal, Shane; Ghorashi, Seyed Ali

    2018-03-01

    Campylobacter infection is a common cause of bacterial gastroenteritis in humans and remains a significant global public health issue. The capability of two multiplex PCR (mPCR)-high-resolution melt (HRM) curve analysis methods (i.e., mPCR1-HRM and mPCR2-HRM) to detect and differentiate 24 poultry isolates and three reference strains of Campylobacter jejuni and Campylobacter coli was investigated. Campylobacter jejuni and C. coli were successfully differentiated in both assays, but the differentiation power of mPCR2-HRM targeting the cadF gene was found superior to that of mPCR1-HRM targeting the gpsA gene or a hypothetical protein gene. However, higher intraspecies variation within C. coli and C. jejuni isolates was detected in mPCR1-HRM when compared with mPCR2-HRM. Both assays were rapid and required minimum interpretation skills for discrimination between and within Campylobacter species when using HRM curve analysis software.

  15. Effect of Feed Form and Whole Grain Feeding on Gastrointestinal Weight and the Prevalence of Campylobacter jejuni in Broilers Orally Infected

    PubMed Central

    Gracia, Marta Isabel; Sánchez, Jaime; Millán, Carlos; Casabuena, Óscar; Vesseur, Peter; Martín, Ángel; García-Peña, Francisco Javier; Medel, Pedro

    2016-01-01

    Two independent trials were carried out to evaluate the effect of feed form, whole wheat (WW) and oat hulls (OH) addition on gastrointestinal (GIT) weight and Campylobacter jejuni colonization in orally infected birds. In Trial 1, there were six treatments factorially arranged with two feed forms (mash vs pellets), and three levels of WW from 1-21/22-42d: 0/0, 7.5/15%, 15/30%. Broilers were allocated in cages (3 birds/cage, 12 cages/treatment). In Trial 2, there were three treatments: a mash diet, a mash diet including WW (7.5% from 1–21 and 15% from 22-42d), and a third treatment including also 5%OH. Broilers were allocated in floor pens (1 pen with 30 birds/treatment). At 14d, all broilers in Trial 1 or 3 broilers/pen in Trial 2 were orally challenged with 1.5 x 105 cfu of C. jejuni ST-45 /. In Trial 1, birds fed pelleted diets consumed 13.5% more feed, gained 31% more weight, and presented 12.9% better feed conversion for the whole trial (P<0.05). Pelleting decreased the relative weight of GIT and gizzard and increased the relative weight of proventriculus (P<0.05). Mash diets decreased pH in the gizzard (P<0.05). Inclusion of WW decreased the relative weight of proventriculus, increased gizzard weight, and reduced pH in the gizzard (P<0.05). At 21d of age, mash tended to reduce C. jejuni compared to pellets (7.85 vs 8.27 log10cfu/g; P = 0.091) and WW inclusion at 7.5/15% reduced C. jejuni colonization when compared to lower and higher inclusion (P<0.05). In Trial 2, birds fed T3 (WW+OH) showed 1.38 log10cfu/g less than birds fed Control diet (P<0.05). In conclusion, despite of the clear morphological changes in the GIT derived of FF and WW inclusion, no clear reductions in C. jejuni populations in the ceca were observed. However, WW and OH inclusion to mash diets significantly reduced cecal C. jejuni colonization at 42 days. PMID:27500730

  16. Discrimination of Major Capsular Types of Campylobacter jejuni by Multiplex PCR

    DTIC Science & Technology

    2011-05-01

    strains of known Pcnnt:r type No. ,,f s1r.1ins with !he following result: %" 11 rimcr set Tl•tat TrUt: pu~illVC: Fat ~~ pus it iw F:.tsc u~gativc Trm...D., B. Kuzniar. B. Shames . L. :\\, Kurjunczyk, and .J, L. Penner. 1992. Variation of the 0 antigen of Campyh•hacter jejuni in viv’’· J. Med

  17. Affinity Probe Capillary Electrophoresis Evaluation of Aptamer Binding to Campylobacter jejuni Bacteria

    DTIC Science & Technology

    2009-11-01

    absorption coefficients (260nm) of 173,100 M cm–1. Desired stock solutions were freshly prepared with tris- borate ethylenediaminetetraacetic acid (EDTA... McMasters , and Paul M. Pellegrino ARL-TR-5015 November 2009 Approved for public release...Aptamer Binding to Campylobacter jejuni Bacteria Dimitra N. Stratis-Cullum, Sun McMasters , and Paul M. Pellegrino Sensors and Electron Devices

  18. Characterization and Reactivity of Broiler Chicken Sera to Selected Recombinant Campylobacter jejuni Chemotactic Proteins

    USDA-ARS?s Scientific Manuscript database

    Campylobacter jejuni, a Gram-negative rod bacterium, is the leading causative agent of human acute bacterial gastroenteritis worldwide. Consumption and handling of raw or undercooked poultry are regarded as a major source for human infection. Because bacterial chemotaxis guides microorganisms to c...

  19. Selenium-Dependent Biogenesis of Formate Dehydrogenase in Campylobacter jejuni Is Controlled by the fdhTU Accessory Genes

    PubMed Central

    Shaw, Frances L.; Mulholland, Francis; Le Gall, Gwénaëlle; Porcelli, Ida; Hart, Dave J.; Pearson, Bruce M.

    2012-01-01

    The food-borne bacterial pathogen Campylobacter jejuni efficiently utilizes organic acids such as lactate and formate for energy production. Formate is rapidly metabolized via the activity of the multisubunit formate dehydrogenase (FDH) enzyme, of which the FdhA subunit is predicted to contain a selenocysteine (SeC) amino acid. In this study we investigated the function of the cj1500 and cj1501 genes of C. jejuni, demonstrate that they are involved in selenium-controlled production of FDH, and propose the names fdhT and fdhU, respectively. Insertional inactivation of fdhT or fdhU in C. jejuni resulted in the absence of FdhA and FdhB protein expression, reduced fdhABC RNA levels, the absence of FDH enzyme activity, and the lack of formate utilization, as assessed by 1H nuclear magnetic resonance. The fdhABC genes are transcribed from a single promoter located two genes upstream of fdhA, and the decrease in fdhABC RNA levels in the fdhU mutant is mediated at the posttranscriptional level. FDH activity and the ability to utilize formate were restored by genetic complementation with fdhU and by supplementation of the growth media with selenium dioxide. Disruption of SeC synthesis by inactivation of the selA and selB genes also resulted in the absence of FDH activity, which could not be restored by selenium supplementation. Comparative genomic analysis suggests a link between the presence of selA and fdhTU orthologs and the predicted presence of SeC in FdhA. The fdhTU genes encode accessory proteins required for FDH expression and activity in C. jejuni, possibly by contributing to acquisition or utilization of selenium. PMID:22609917

  20. Complete genomic sequences of Campylobacter jejuni strains RM3196 (233.94) and RM3197 (308.95) that were isolated from patients with Guillain-Barré Syndrome

    USDA-ARS?s Scientific Manuscript database

    An infection with Campylobacter jejuni subsp. jejuni (Cjj) is a leading cause of foodborne gastroenteritis in humans and also the most prevalent infection preceding Guillain-Barré syndrome (GBS). This study describes the complete genomic sequences of Cjj HS:41 strains RM3196 (233.94) and RM3197 (308...

  1. Fundamental Boiling and RP-1 Freezing Experiments

    NASA Technical Reports Server (NTRS)

    Goode, Brian

    2002-01-01

    The prestart thermal conditioning of the hardware in LOX (liquid oxygen) systems involve heat transfer between LOX and metal where boiling plays a large role. Information is easily found on nucleate boiling, maximum heat flux, minimum heat flux and film boiling for common fluids like water. After looking at these standard correlations it was felt more data was needed for the cool down side transition boiling for the LN2 and LOX. In particular interest is the film boiling values, the temperature at which transition begins and the slope as peak heat flux is approached. The ultimate goal is an array of boiling heat transfer coefficient as a function of surface temperature which can be used in the chilldown model of the feed system, engine and bleed system for X-34. The first experiment consisted of an actual MC-1 LOX Impeller which had been machined backwards, that was instrumented with 17 surface thermocouples and submerged in liquid nitrogen. The thermocouples were installed on metal thicknesses varying from the thin inducer to the thick hub.

  2. Boiling Experiment Facility for Heat Transfer Studies in Microgravity

    NASA Technical Reports Server (NTRS)

    Delombard, Richard; McQuillen, John; Chao, David

    2008-01-01

    Pool boiling in microgravity is an area of both scientific and practical interest. By conducting tests in microgravity, it is possible to assess the effect of buoyancy on the overall boiling process and assess the relative magnitude of effects with regards to other "forces" and phenomena such as Marangoni forces, liquid momentum forces, and microlayer evaporation. The Boiling eXperiment Facility is now being built for the Microgravity Science Glovebox that will use normal perfluorohexane as a test fluid to extend the range of test conditions to include longer test durations and less liquid subcooling. Two experiments, the Microheater Array Boiling Experiment and the Nucleate Pool Boiling eXperiment will use the Boiling eXperiment Facility. The objectives of these studies are to determine the differences in local boiling heat transfer mechanisms in microgravity and normal gravity from nucleate boiling, through critical heat flux and into the transition boiling regime and to examine the bubble nucleation, growth, departure and coalescence processes. Custom-designed heaters will be utilized to achieve these objectives.

  3. A large outbreak of Campylobacter jejuni infection in a university college caused by chicken liver pâté, Australia, 2013.

    PubMed

    Moffatt, C R M; Greig, A; Valcanis, M; Gao, W; Seemann, T; Howden, B P; Kirk, M D

    2016-10-01

    In October 2013, public health authorities were notified of a suspected outbreak of gastroenteritis in students and guests following a catered function at a university residential college. A retrospective cohort study was undertaken to examine whether foods served at the function caused illness. A total of 56 cases of gastroenteritis, including seven laboratory-confirmed cases of Campylobacter jejuni infection, were identified in 235 eligible respondents. Univariate analysis showed a significant association with a chicken liver pâté entrée [relative risk (RR) 3·64, 95% confidence interval (CI) 2·03-6·52, P < 0·001], which retained significance after adjustment for confounding via multivariable analysis (adjusted RR 2·80, 95% CI 1·26-6·19, P = 0·01). C. jejuni and C. coli were also isolated in chicken liver pâté recovered from the college's kitchen. Subsequent whole genome multilocus sequence typing (wgMLST) of clinical and food-derived C. jejuni isolates showed three genetically distinct sequence types (STs) comprising ST528, ST535 (both clinically derived) and ST991 (food derived). The study demonstrates the value of utilizing complementary sources of evidence, including genomic data, to support public health investigations. The use of wgMLST highlights the potential for significant C. jejuni diversity in epidemiologically related human and food isolates recovered during outbreaks linked to poultry liver.

  4. Boiling enriches the linear polysulfides and the hydrogen sulfide-releasing activity of garlic.

    PubMed

    Tocmo, Restituto; Wu, Yuchen; Liang, Dong; Fogliano, Vincenzo; Huang, Dejian

    2017-04-15

    Garlic is rich in polysulfides, and some of them can be H 2 S donors. This study was conducted to explore the effect of cooking on garlic's organopolysulfides and H 2 S-releasing activity. Garlic bulbs were crushed and boiled for a period ranging from 3 to 30min and the solvent extracts were analyzed by GC-MS/FID and HPLC. A cell-based assay was used to measure the H 2 S-releasing activity of the extracts. Results showed that the amounts of allyl polysulfides increased in crushed garlic boiled for 6-10min; however, prolonging the thermal treatment to 20 or 30min decreased their concentrations. Data of the H 2 S-releasing activity, expressed as diallyl trisulfide equivalents (DATS-E), parallel this trend, being significantly higher at 6 and 10min boiling. Our results showed enhancement of H 2 S-releasing activity upon moderate boiling, suggesting that shorter cooking time may maximize its health benefits as a dietary source of natural H 2 S donors. Copyright © 2016 Elsevier Ltd. All rights reserved.

  5. Pool and flow boiling in variable and microgravity

    NASA Technical Reports Server (NTRS)

    Merte, Herman, Jr.

    1994-01-01

    As is well known, boiling is an effective mode of heat transfer in that high heat flux levels are possible with relatively small temperature differences. Its optimal application requires that the process be adequately understood. A measure of the understanding of any physical event lies in the ability to predict its behavior in terms of the relevant parameters. Despite many years of research the predictability of boiling is currently possible only for quite specialized circumstances, e.g., the critical heat flux and film boiling for the pool boiling case, and then only with special geometries. Variable gravity down to microgravity provides the opportunity to test this understanding, but possibly more important, by changing the dimensional and time scales involved permits more detailed observations of elements involved in the boiling process, and perhaps discloses phenomena heretofore unknown. The focus here is on nucleate boiling although, as will be demonstrated below, under but certain circumstances in microgravity it can take place concurrently with the dryout process. In the presence of earth gravity or forced convection effects, the latter process is usually referred to as film boiling. However, no vapor film as such forms with pool boiling in microgravity, only dryout. Initial results are presented here for pool boiling in microgravity, and were made possible at such an early date by the availability of the Get-Away-Specials (GAS). Also presented here are some results of ground testing of a flow loop for the study of low velocity boiling, eventually to take place also in microgravity. In the interim, variable buoyancy normal to the heater surface is achieved by rotation of the entire loop relative to earth gravity. Of course, this is at the expense of varying the buoyancy parallel to the heater surface. Two questions which must be resolved early in the study of flow boiling in microgravity are (1) the lower limits of liquid flow velocity where buoyancy

  6. Plant-derived antimicrobial eugenol modulates C. jejuni proteome and virulence critical for colonization in chickens

    USDA-ARS?s Scientific Manuscript database

    Campylobacter jejuni is an important foodborne pathogen that causes severe diarrhea in humans. Chickens act as the reservoir host for Campylobacter, wherein the pathogen colonizes the ceca leading to contaminated poultry products during slaughter. The potential of natural intervention strategies, in...

  7. Functional Analysis of the RdxA and RdxB Nitroreductases of Campylobacter jejuni Reveals that Mutations in rdxA Confer Metronidazole Resistance▿ †

    PubMed Central

    Ribardo, Deborah A.; Bingham-Ramos, Lacey K.; Hendrixson, David R.

    2010-01-01

    Campylobacter jejuni is a leading cause of gastroenteritis in humans and a commensal bacterium of the intestinal tracts of many wild and agriculturally significant animals. We identified and characterized a locus, which we annotated as rdxAB, encoding two nitroreductases. RdxA was found to be responsible for sensitivity to metronidazole (Mtz), a common therapeutic agent for another epsilonproteobacterium, Helicobacter pylori. Multiple, independently derived mutations in rdxA but not rdxB resulted in resistance to Mtz (Mtzr), suggesting that, unlike the case in H. pylori, Mtzr might not be a polygenic trait. Similarly, Mtzr C. jejuni was isolated after both in vitro and in vivo growth in the absence of selection that contained frameshift, point, insertion, or deletion mutations within rdxA, possibly revealing genetic variability of this trait in C. jejuni due to spontaneous DNA replication errors occurring during normal growth of the bacterium. Similar to previous findings with H. pylori RdxA, biochemical analysis of C. jejuni RdxA showed strong oxidase activity, with reduction of Mtz occurring only under anaerobic conditions. RdxB showed similar characteristics but at levels lower than those for RdxA. Genetic analysis confirmed that rdxA and rdxB are cotranscribed and induced during in vivo growth in the chick intestinal tract, but an absence of these genes did not strongly impair C. jejuni for commensal colonization. Further studies indicate that rdxA is a convenient locus for complementation of mutants in cis. Our work contributes to the growing knowledge of determinants contributing to susceptibility to Mtz (Mtzs) and supports previous observations of the fundamental differences in the activities of nitroreductases from epsilonproteobacteria. PMID:20118248

  8. Characterizing preferential groundwater discharge through boils using temperature

    NASA Astrophysics Data System (ADS)

    Vandenbohede, A.; de Louw, P. G. B.; Doornenbal, P. J.

    2014-03-01

    In The Netherlands, preferential groundwater discharge trough boils is a key process in the salinization of deep polders. Previous work showed that boils also influence the temperature in the subsurface and of surface water. This paper elaborates on this process combining field observations with numerical modeling. As is the case for salinity, a distinct anomaly in the subsurface and surface water temperature can be attributed to boils. Lines of equal temperature are distorted towards the boil, which can be considered as an upconing of the temperature profile by analogy of the upconing of a fresh-saltwater interface. The zone of this distortion is limited to the immediate vicinity of the boil, being about 5 m in the aquitard which holds the boil's conduit, or maximum a few dozens of meters in the underlying aquifer. In the aquitard, heat transport is conduction dominated whereas this is convection dominated in the aquifer. The temperature anomaly differs from the salinity anomaly by the smaller radius of influence and faster time to reach a new steady-state of the former. Boils discharge water with a temperature equal to the mean groundwater temperature. This influences the yearly and diurnal variation of ditch water temperature in the immediate vicinity of the boil importantly but also the temperature in the downstream direction. Temporary nature of the boil (e.g. stability of the conduit, discharge rate), uncertainty on the 3D construction of the conduit and heterogeneity of the subsoil make it unlikely that temperature measurements can be interpreted further than a qualitative level.

  9. The extent of surface contamination of retailed chickens with Campylobacter jejuni serogroups.

    PubMed Central

    Hood, A. M.; Pearson, A. D.; Shahamat, M.

    1988-01-01

    Eighty-two chickens purchased at 11 retailers (supplied by 12 wholesalers) in the south of England were cultured for Campylobacter jejuni by a method involving total immersion. The organism was isolated from 22 (48%) of 46 fresh birds, 12 of 12 uneviscerated (New York dressed) birds, but only 1 of 24 frozen birds. Viable counts of up to 1.5 x 10(6)/chicken were obtained from fresh birds and 2.4 x 10(7)/chicken from uneviscerated birds. Surface swabbing of breasts, thighs, wings and vents of fresh chickens showed that contamination was generally distributed over the carcasses. Salmonellas were found in only 2 of 69 of the fresh chickens. The prevalence of several Lior and Penner C. jejuni serogroups was similar in chickens and sporadic human cases of enteritis. Penner serogroup 4 (mostly of Lior serogroup 1) was found in 26% of human isolates and 14% of chicken isolates. The rising incidence of campylobacter enteritis during the last 6 years could well be a reflection of the increasing proportion of fresh chickens consumed over that period (32% higher in 1986 than in 1981). PMID:3338503

  10. Structural changes of malt proteins during boiling.

    PubMed

    Jin, Bei; Li, Lin; Liu, Guo-Qin; Li, Bing; Zhu, Yu-Kui; Liao, Liao-Ning

    2009-03-09

    Changes in the physicochemical properties and structure of proteins derived from two malt varieties (Baudin and Guangmai) during wort boiling were investigated by differential scanning calorimetry, SDS-PAGE, two-dimensional electrophoresis, gel filtration chromatography and circular dichroism spectroscopy. The results showed that both protein content and amino acid composition changed only slightly during boiling, and that boiling might cause a gradual unfolding of protein structures, as indicated by the decrease in surface hydrophobicity and free sulfhydryl content and enthalpy value, as well as reduced alpha-helix contents and markedly increased random coil contents. It was also found that major component of both worts was a boiling-resistant protein with a molecular mass of 40 kDa, and that according to the two-dimensional electrophoresis and SE-HPLC analyses, a small amount of soluble aggregates might be formed via hydrophobic interactions. It was thus concluded that changes of protein structure caused by boiling that might influence beer quality are largely independent of malt variety.

  11. A carvacrol wash and/or a chitosan based coating reduced Campylobacter jejuni on chicken wingettes

    USDA-ARS?s Scientific Manuscript database

    Campylobacter jejuni is a leading cause of foodborne disease in humans, largely associated with consumption of contaminated poultry and poultry products. With increasing consumer demand for natural and minimally processed foods, the use of Generally Recognized as Safe (GRAS) status plant derived com...

  12. Ciliate ingestion and digestion: flow cytometric measurements and regrowth of a digestion-resistant campylobacter jejuni

    USDA-ARS?s Scientific Manuscript database

    We developed a method to measure ingestion and digestion rates of bacterivorous protists feeding on pathogenic bacteria. We tested this method using the enteric bacteria Campylobacter jejuni and a freshwater colpodid ciliate. Campylobacter and a non-pathogenic bacteria isolated from the environment ...

  13. A review on boiling heat transfer enhancement with nanofluids

    PubMed Central

    2011-01-01

    There has been increasing interest of late in nanofluid boiling and its use in heat transfer enhancement. This article covers recent advances in the last decade by researchers in both pool boiling and convective boiling applications, with nanofluids as the working fluid. The available data in the literature is reviewed in terms of enhancements, and degradations in the nucleate boiling heat transfer and critical heat flux. Conflicting data have been presented in the literature on the effect that nanofluids have on the boiling heat-transfer coefficient; however, almost all researchers have noted an enhancement in the critical heat flux during nanofluid boiling. Several researchers have observed nanoparticle deposition at the heater surface, which they have related back to the critical heat flux enhancement. PMID:21711794

  14. The food-borne pathogen Campylobacter jejuni responds to the bile salt deoxycholate with countermeasures to reactive oxygen species

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Negretti, Nicholas M.; Gourley, Christopher R.; Clair, Geremy

    In this study, bile plays an important role in digestion, absorption of fats, and the excretion of waste products, while concurrently providing a critical barrier against colonization by harmful bacteria. Previous studies have demonstrated that gut pathogens react to bile by adapting their protein synthesis. The ability of pathogens to respond to bile is remarkably complex and still incompletely understood. Here we show that Campylobacter jejuni, a leading bacterial cause of human diarrheal illness worldwide, responds to deoxycholate, a component of bile, by altering global gene transcription in a manner consistent with a strategy to mitigate exposure to reactive oxygenmore » stress. More specifically, continuous growth of C. jejuni in deoxycholate was found to: induce the production of reactive oxygen species (ROS); decrease succinate dehydrogenase activity (complex II of the electron transport chain); increase catalase activity that is involved in H 2O 2 breakdown; and result in DNA strand breaks. Congruently, by adding 4-hydroxy-TEMPO (TEMPOL), a superoxide dismutase mimic, that reacts with superoxide to cultures under deoxycholate-mediated ROS stress, C. jejuni growth in the presence of deoxycholate was rescued. We postulate that continuous exposure of a number of enteric pathogens to deoxycholate stimulates a conserved survival response to this stressor.« less

  15. The food-borne pathogen Campylobacter jejuni responds to the bile salt deoxycholate with countermeasures to reactive oxygen species

    DOE PAGES

    Negretti, Nicholas M.; Gourley, Christopher R.; Clair, Geremy; ...

    2017-11-13

    In this study, bile plays an important role in digestion, absorption of fats, and the excretion of waste products, while concurrently providing a critical barrier against colonization by harmful bacteria. Previous studies have demonstrated that gut pathogens react to bile by adapting their protein synthesis. The ability of pathogens to respond to bile is remarkably complex and still incompletely understood. Here we show that Campylobacter jejuni, a leading bacterial cause of human diarrheal illness worldwide, responds to deoxycholate, a component of bile, by altering global gene transcription in a manner consistent with a strategy to mitigate exposure to reactive oxygenmore » stress. More specifically, continuous growth of C. jejuni in deoxycholate was found to: induce the production of reactive oxygen species (ROS); decrease succinate dehydrogenase activity (complex II of the electron transport chain); increase catalase activity that is involved in H 2O 2 breakdown; and result in DNA strand breaks. Congruently, by adding 4-hydroxy-TEMPO (TEMPOL), a superoxide dismutase mimic, that reacts with superoxide to cultures under deoxycholate-mediated ROS stress, C. jejuni growth in the presence of deoxycholate was rescued. We postulate that continuous exposure of a number of enteric pathogens to deoxycholate stimulates a conserved survival response to this stressor.« less

  16. Modification of the Campylobacter jejuni flagellin glycan by the product of the Cj1295 homopolymeric-tract-containing gene

    PubMed Central

    Hitchen, Paul; Brzostek, Joanna; Panico, Maria; Butler, Jonathan A.; Morris, Howard R.; Dell, Anne; Linton, Dennis

    2010-01-01

    The Campylobacter jejuni flagellin protein is O-glycosylated with structural analogues of the nine-carbon sugar pseudaminic acid. The most common modifications in the C. jejuni 81-176 strain are the 5,7-di-N-acetylated derivative (Pse5Ac7Ac) and an acetamidino-substituted version (Pse5Am7Ac). Other structures detected include O-acetylated and N-acetylglutamine-substituted derivatives (Pse5Am7Ac8OAc and Pse5Am7Ac8GlnNAc, respectively). Recently, a derivative of pseudaminic acid modified with a di-O-methylglyceroyl group was detected in C. jejuni NCTC 11168 strain. The gene products required for Pse5Ac7Ac biosynthesis have been characterized, but those genes involved in generating other structures have not. We have demonstrated that the mobility of the NCTC 11168 flagellin protein in SDS-PAGE gels can vary spontaneously and we investigated the role of single nucleotide repeats or homopolymeric-tract-containing genes from the flagellin glycosylation locus in this process. One such gene, Cj1295, was shown to be responsible for structural changes in the flagellin glycoprotein. Mass spectrometry demonstrated that the Cj1295 gene is required for glycosylation with the di-O-methylglyceroyl-modified version of pseudaminic acid. PMID:20338909

  17. Transition boiling heat transfer and the film transition regime

    NASA Technical Reports Server (NTRS)

    Ramilison, J. M.; Lienhard, J. H.

    1987-01-01

    The Berenson (1960) flat-plate transition-boiling experiment has been recreated with a reduced thermal resistance in the heater, and an improved access to those portions of the transition boiling regime that have a steep negative slope. Tests have been made in Freon-113, acetone, benzene, and n-pentane boiling on horizontal flat copper heaters that have been mirror-polished, 'roughened', or teflon-coated. The resulting data reproduce and clarify certain features observed by Berenson: the modest surface finish dependence of boiling burnout, and the influence of surface chemistry on both the minimum heat flux and the mode of transition boiling, for example. A rational scheme of correlation yields a prediction of the heat flux in what Witte and Lienhard (1982) previously identified as the 'film-transition boiling' region. It is also shown how to calculate the heat flux at the boundary between the pure-film, and the film-transition, boiling regimes, as a function of the advancing contact angle.

  18. The use of FTA cards for transport and detection of gyrA mutation of Campylobacter jejuni from poultry.

    PubMed

    Sierra-Arguello, Y M; Faulkner, O; Tellez, G; Hargis, B M; Pinheiro do Nascimento, V

    2016-04-01

    The purpose of the present study was to evaluate a technique involving the use of commercially available FTA classic card (Whatman) for transporting and detection of DNA to use in PCR analysis and genetic sequencing of Campylobacter jejuni of poultry origin. Fifty isolates of Campylobacter jejuni were obtained from broiler carcasses in Rio Grande do Sul, Brazil. Antimicrobial susceptibility testing to ciprofloxacin revealed that all 50 isolates were resistant to ciprofloxacin. Each isolate was transferred to Brucella broth tubes and incubated overnight at 41.5°C. Cell cultures were diluted to match a McFarland Turbidity Standard 0.5, and 110 μL of the cell suspension were applied to one circle on Whatman FTA classic cards. The samples were then covered and allowed to dry at room temperature. Cards were identified and stored at room temperature until further use (3 mo after collection). FTA cards were shipped for analysis to the Department of Poultry Science, University of Arkansas. Amplification of the Campylobacter gyrA gene was successful and demonstrated strong bands for a large amplicon for all 50 samples preserved on FTA cards. Mutations present in each gene were confirmed by DNA sequencing. Then, 7 samples were chosen for the sequencing. The detection of a mutation regarding ciprofloxacin-resistant isolates revealed that 7 samples had a mutation in the gyrA gene. In conclusion, the characteristics of the profiles suggest that the DNA has maintained its integrity after 3 mo of storage at room temperature and is a suitable template for PCR and sequencing from Campylobacter samples. The application of this technology has potential in numerous methodologies, especially when working in remote areas and in developing countries where access to laboratory facilities and equipment is limited. © 2016 Poultry Science Association Inc.

  19. Novel plasmid conferring kanamycin and tetracycline resistance in turkey-derived Campylobacter jejuni strain 11601MD

    USDA-ARS?s Scientific Manuscript database

    In Campylobacter spp., resistance to the antibiotics kanamycin and tetracycline is frequently associated with plasmid-borne genes. However, relatively few plasmids of Campylobacter jejuni have been fully characterized to date. A novel plasmid (p11601MD; 44,095 bp.) harboring tet(O) was identified in...

  20. Cleaning and disinfection programs against Campylobacter jejuni for broiler chickens: productive performance, microbiological assessment and characterization1

    PubMed Central

    Deliberali Lelis, Karoline; Granghelli, Carlos Alexandre; Carão de Pinho, Agatha Cristina; Ribeiro Almeida Queiroz, Sabrina; Fernandes, Andrezza Maria; Moro de Souza, Ricardo Luiz; Gaglianone Moro, Maria Estela; de Andrade Bordin, Roberto; de Albuquerque, Ricardo

    2017-01-01

    Abstract Detailed cleaning and disinfection programs aims to reduce infection pressure from microorganisms from one flock to the next. However, studies evaluating the benefits to poultry performance, the sanitary status of the facilities, and the sanitary quality of the meat are rarely found. Thus, this study was designed to evaluate 2 cleaning and disinfecting programs regarding their influence on productive performance, elimination of Campylobacter, and characterization of Campylobacter jejuni strains when applied to broiler chickens’ facilities. Two subsequent flocks with 960 birds each were distributed into 32 pens containing 30 birds each. In the first, the whole flock was inoculated with a known strain of Campylobacter jejuni in order to contaminate the environment. In the second flock, performance and microbiological evaluations were done, characterizing an observational study between 2 cleaning and disinfection programs, regular and proposed. The regular program consisted of sweeping facilities, washing equipment and environment with water and neutral detergent. The proposed cleaning program consisted of dry and wet cleaning, application of 2 detergents (one acid and one basic) and 2 disinfectants (250 g/L glutaraldehyde and 185 g/L formaldehyde at 0.5% and 210 g/L para-chloro-meta-cresol at 4%). Total microorganism count in the environment and Campylobacter spp. identification were done for the microbiological assessment of the environment and carcasses. The positive samples were submitted to molecular identification of Campylobacter spp. and posterior genetic sequencing of the species identified as Campylobacter jejuni. The birds housed in the facilities and submitted to the proposed treatment had better performance when compared to the ones in the regular treatment, most likely because there was a smaller total microorganism count on the floor, walls, feeders and drinkers. The proposed program also resulted in a reduction of Campylobacter spp. on

  1. Gene Loss and Lineage-Specific Restriction-Modification Systems Associated with Niche Differentiation in the Campylobacter jejuni Sequence Type 403 Clonal Complex

    PubMed Central

    Morley, Laura; McNally, Alan; Paszkiewicz, Konrad; Corander, Jukka; Méric, Guillaume; Sheppard, Samuel K.; Blom, Jochen

    2015-01-01

    Campylobacter jejuni is a highly diverse species of bacteria commonly associated with infectious intestinal disease of humans and zoonotic carriage in poultry, cattle, pigs, and other animals. The species contains a large number of distinct clonal complexes that vary from host generalist lineages commonly found in poultry, livestock, and human disease cases to host-adapted specialized lineages primarily associated with livestock or poultry. Here, we present novel data on the ST403 clonal complex of C. jejuni, a lineage that has not been reported in avian hosts. Our data show that the lineage exhibits a distinctive pattern of intralineage recombination that is accompanied by the presence of lineage-specific restriction-modification systems. Furthermore, we show that the ST403 complex has undergone gene decay at a number of loci. Our data provide a putative link between the lack of association with avian hosts of C. jejuni ST403 and both gene gain and gene loss through nonsense mutations in coding sequences of genes, resulting in pseudogene formation. PMID:25795671

  2. Water boiling inside carbon nanotubes: toward efficient drug release.

    PubMed

    Chaban, Vitaly V; Prezhdo, Oleg V

    2011-07-26

    We show using molecular dynamics simulation that spatial confinement of water inside carbon nanotubes (CNTs) substantially increases its boiling temperature and that a small temperature growth above the boiling point dramatically raises the inside pressure. Capillary theory successfully predicts the boiling point elevation down to 2 nm, below which large deviations between the theory and atomistic simulation take place. Water behaves qualitatively different inside narrow CNTs, exhibiting transition into an unusual phase, where pressure is gas-like and grows linearly with temperature, while the diffusion constant is temperature-independent. Precise control over boiling by CNT diameter, together with the rapid growth of inside pressure above the boiling point, suggests a novel drug delivery protocol. Polar drug molecules are packaged inside CNTs; the latter are delivered into living tissues and heated by laser. Solvent boiling facilitates drug release.

  3. Serologic Evidence of Previous Campylobacter jejuni Infection in Patients with the Guillain-Barre Syndrome

    DTIC Science & Technology

    1993-06-15

    chronic inflammatory demyelinating polyneuropathy , and polyneuropathy associated with IgM paraproteinemia. creased the sensitivity but improved the...paraproteine- were employees of or visitors to the Infectious Diseases Divi- ,,i- hronic inflammatory demyelinating polyneuropathy , sion of Vanderbilt... polyneuropathy ," is an inflammatory de- jejuni infection before onset of neurologic symptoms. myelinating disease of peripheral nerves characterized

  4. Cleaning and disinfection programs against Campylobacter jejuni for broiler chickens: productive performance, microbiological assessment and characterization.

    PubMed

    Castro Burbarelli, Maria Fernanda de; do Valle Polycarpo, Gustavo; Deliberali Lelis, Karoline; Granghelli, Carlos Alexandre; Carão de Pinho, Agatha Cristina; Ribeiro Almeida Queiroz, Sabrina; Fernandes, Andrezza Maria; Moro de Souza, Ricardo Luiz; Gaglianone Moro, Maria Estela; de Andrade Bordin, Roberto; de Albuquerque, Ricardo

    2017-09-01

    Detailed cleaning and disinfection programs aims to reduce infection pressure from microorganisms from one flock to the next. However, studies evaluating the benefits to poultry performance, the sanitary status of the facilities, and the sanitary quality of the meat are rarely found. Thus, this study was designed to evaluate 2 cleaning and disinfecting programs regarding their influence on productive performance, elimination of Campylobacter, and characterization of Campylobacter jejuni strains when applied to broiler chickens' facilities. Two subsequent flocks with 960 birds each were distributed into 32 pens containing 30 birds each. In the first, the whole flock was inoculated with a known strain of Campylobacter jejuni in order to contaminate the environment. In the second flock, performance and microbiological evaluations were done, characterizing an observational study between 2 cleaning and disinfection programs, regular and proposed. The regular program consisted of sweeping facilities, washing equipment and environment with water and neutral detergent. The proposed cleaning program consisted of dry and wet cleaning, application of 2 detergents (one acid and one basic) and 2 disinfectants (250 g/L glutaraldehyde and 185 g/L formaldehyde at 0.5% and 210 g/L para-chloro-meta-cresol at 4%). Total microorganism count in the environment and Campylobacter spp. identification were done for the microbiological assessment of the environment and carcasses. The positive samples were submitted to molecular identification of Campylobacter spp. and posterior genetic sequencing of the species identified as Campylobacter jejuni. The birds housed in the facilities and submitted to the proposed treatment had better performance when compared to the ones in the regular treatment, most likely because there was a smaller total microorganism count on the floor, walls, feeders and drinkers. The proposed program also resulted in a reduction of Campylobacter spp. on floors

  5. Boiling on Microconfigured Composite Surfaces Enhanced

    NASA Technical Reports Server (NTRS)

    Chao, David F.

    2000-01-01

    Boiling heat transfer is one of the key technologies for the two-phase active thermal-control system used on space platforms, as well as for the dynamic power systems aboard the International Space Station. Because it is an effective heat transfer mode, boiling is integral to many space applications, such as heat exchangers and other cooling devices. Nucleate boiling near the critical heat flux (CHF) can transport very large thermal loads with a much smaller device and much lower pumping power than for single-phase heat exchangers. However, boiling performance sharply deteriorates in a reduced-gravity environment, and operation in the CHF regime is somewhat perilous because of the risk of burnout to the device surface. New materials called microconfigured metal-graphite composites can enhance boiling. The photomicrograph shows the microconfiguration (x3000) of the copper-graphite (Cu-Gr) surface as viewed by scanning electronic microscope. The graphite fiber tips appear as plateaus with rugged surfaces embedded in the copper matrix. It has been experimentally demonstrated that this type of material manifests excellent boiling heat transfer performance characteristics and an increased CHF. Nonisothermal surfaces were less sensitive to variations of wall superheat in the CHF regime. Because of the great difference in conductivity between the copper base and the graphite fiber, the composite surfaces have a nonisothermal surface characteristic and, therefore, will have a much larger "safe" operating region in the CHF regime. In addition, the thermocapillary forces induced by the temperature differences between the fiber tips and the metal matrix play an important role in bubble detachment, and may not be adversely affected in a reduced-gravity environment. All these factors indicate that microconfigured composites may improve the reliability and economy (dominant factors in all space applications) of various thermal components found on spacecraft during future

  6. BOILING HEAT TRANSFER IN ZERO GRAVITY

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Zara, E.A.

    1964-01-01

    The preliminary results of a research program to determine the effects of zero and near zero gravity on boiling heat transfer are presented. Zero gravity conditions were obtained on the ASD KC-135 zero gravity test aircraft, capable of providing 30-seconds of zero gravity. Results of the program to date indicate that nucleate (bubble) boiling heat transfer rates are not greatly affected by the absence of gravity forces. However, radical pressure increases were observed that will dictate special design considerations to space vehicle systems utilizing pool boiling processes, such as cryogenic or other fluid storage vessels where thermal input to themore » fluid is used for vessel pressurization. (auth)« less

  7. Enhancements of Nucleate Boiling Under Microgravity Conditions

    NASA Technical Reports Server (NTRS)

    Zhang, Nengli; Chao, David F.; Yang, W. J.

    2000-01-01

    This paper presents two means for enhancing nucleate boiling and critical heat flux under microgravity conditions: using micro-configured metal-graphite composites as the boiling surface and dilute aqueous solutions of long-chain alcohols as the working fluid. In the former, thermocapillary force induced by temperature difference between the graphite-fiber tips and the metal matrix plays an important role in bubble detachment. Thus boiling-heat transfer performance does not deteriorate in a reduced-gravity environment. In the latter cases, the surface tension-temperature gradient of the long-chain alcohol solutions turns positive as the temperature exceeds a certain value. Consequently, the Marangoni effect does not impede, but rather aids in bubble departure from the heating surface. This feature is most favorable in microgravity. As a result, the bubble size of departure is substantially reduced at higher frequencies. Based on the existing experimental data, and a two-tier theoretical model, correlation formulas are derived for nucleate boiling on the copper-graphite and aluminum-graphite composite surfaces, in both the isolated and coalesced bubble regimes. In addition, performance equations for nucleate boiling and critical heat flux in dilute aqueous solutions of long-chain alcohols are obtained.

  8. Structural analysis of PseH, the Campylobacter jejuni N-acetyltransferase involved in bacterial O-linked glycosylation.

    PubMed

    Song, Wan Seok; Nam, Mi Sun; Namgung, Byeol; Yoon, Sung-il

    2015-03-20

    Campylobacter jejuni is a bacterium that uses flagella for motility and causes worldwide acute gastroenteritis in humans. The C. jejuni N-acetyltransferase PseH (cjPseH) is responsible for the third step in flagellin O-linked glycosylation and plays a key role in flagellar formation and motility. cjPseH transfers an acetyl group from an acetyl donor, acetyl coenzyme A (AcCoA), to the amino group of UDP-4-amino-4,6-dideoxy-N-acetyl-β-L-altrosamine to produce UDP-2,4-diacetamido-2,4,6-trideoxy-β-L-altropyranose. To elucidate the catalytic mechanism of cjPseH, crystal structures of cjPseH alone and in complex with AcCoA were determined at 1.95 Å resolution. cjPseH folds into a single-domain structure of a central β-sheet decorated by four α-helices with two continuously connected grooves. A deep groove (groove-A) accommodates the AcCoA molecule. Interestingly, the acetyl end of AcCoA points toward an open space in a neighboring shallow groove (groove-S), which is occupied by extra electron density that potentially serves as a pseudosubstrate, suggesting that the groove-S may provide a substrate-binding site. Structure-based comparative analysis suggests that cjPseH utilizes a unique catalytic mechanism of acetylation that has not been observed in other glycosylation-associated acetyltransferases. Thus, our studies on cjPseH will provide valuable information for the design of new antibiotics to treat C. jejuni-induced gastroenteritis. Copyright © 2015 Elsevier Inc. All rights reserved.

  9. Boils and Carbuncles

    MedlinePlus

    ... body. The spreading infection, commonly known as blood poisoning (sepsis), can lead to infections deep within your body, such as your heart (endocarditis) and bone (osteomyelitis). Prevention It's not always possible to prevent boils, especially ...

  10. Conversion of direct process high-boiling residue to monosilanes

    DOEpatents

    Brinson, Jonathan Ashley; Crum, Bruce Robert; Jarvis, Jr., Robert Frank

    2000-01-01

    A process for the production of monosilanes from the high-boiling residue resulting from the reaction of hydrogen chloride with silicon metalloid in a process typically referred to as the "direct process." The process comprises contacting a high-boiling residue resulting from the reaction of hydrogen chloride and silicon metalloid, with hydrogen gas in the presence of a catalytic amount of aluminum trichloride effective in promoting conversion of the high-boiling residue to monosilanes. The present process results in conversion of the high-boiling residue to monosilanes. At least a portion of the aluminum trichloride catalyst required for conduct of the process may be formed in situ during conduct of the direct process and isolation of the high-boiling residue.

  11. A fundamental study of nucleate pool boiling under microgravity

    NASA Technical Reports Server (NTRS)

    Ervin, Jamie S.; Merte, Herman, Jr.

    1991-01-01

    An experimental study of incipient boiling in short-term microgravity and with a/g = +/- 1 for pool boiling was performed. Calibrated thin gold films sputtered on a smoothly polished quartz surface were used simultaneously for thermal resistance measurements and heating of the boiling surface. The gold films were used for both transient and quasi-steady heating surface temperature measurements. Two test vessels were constructed for precise measurement and control of fluid temperature and pressure: a laboratory pool boiling vessel for the a/g = +/- experiments and a pool boiling vessel designed for the 131 m free-fall in the NASA Lewis Research Center Microgravity Research Facility for the microgravity tests. Measurements included the heater surface temperature, the pressure near the heating surface, and the bulk liquid temperatures. High speed photography was used in the experiments. With high quality microgravity and the measured initial temperature of the quiescent test fluid, R113, the temperature distribution in the liquid at the moment of boiling inception resulting from an imposed step in heat flux is known with a certainty not possible previously. The types of boiling propagation across the large flat heating surface are categorized; the conditions necessary for their occurrence are described. Explosive boiling propagation with a striking pattern of small scale protuberances over the entire vapor mass periphery not observed previously at low heat flux levels is described. For the heater surface with a/g = -1, a step in the heater surface temperature of short duration was imposed. The resulting liquid temperature distribution at the moment of boiling inception was different from that obtained with a step in heat flux.

  12. Explosive Boiling at Very Low Heat Fluxes: A Microgravity Phenomenon

    NASA Technical Reports Server (NTRS)

    Hasan, M. M.; Lin, C. S.; Knoll, R. H.; Bentz, M. D.

    1993-01-01

    The paper presents experimental observations of explosive boiling from a large (relative to bubble sizes) flat heating surface at very low heat fluxes in microgravity. The explosive boiling is characterized as either a rapid growth of vapor mass over the entire heating surface due to the flashing of superheated liquid or a violent boiling spread following the appearance of single bubbles on the heating surface. Pool boiling data with saturated Freon 113 was obtained in the microgravity environment of the space shuttle. The unique features of the experimental results are the sustainability of high liquid superheat for long periods and the occurrence of explosive boiling at low heat fluxes (0.2 to 1.2 kW/sq m). For a heat flux of 1.0 kW/sq m a wall superheat of 17.9 degrees C was attained in ten minutes of heating. This was followed by an explosive boiling accompanied with a pressure spike and a violent bulk liquid motion. However, at this heat flux the vapor blanketing the heating surface could not be sustained. Stable nucleate boiling continued following the explosive boiling.

  13. Increased risk for Campylobacter jejuni and C. coli infection of pet origin in dog owners and evidence for genetic association between strains causing infection in humans and their pets.

    PubMed

    Mughini Gras, L; Smid, J H; Wagenaar, J A; Koene, M G J; Havelaar, A H; Friesema, I H M; French, N P; Flemming, C; Galson, J D; Graziani, C; Busani, L; VAN Pelt, W

    2013-12-01

    We compared Campylobacter jejuni/coli multilocus sequence types (STs) from pets (dogs/cats) and their owners and investigated risk factors for pet-associated human campylobacteriosis using a combined source-attribution and case-control analysis. In total, 132/687 pet stools were Campylobacter-positive, resulting in 499 strains isolated (320 C. upsaliensis/helveticus, 100 C. jejuni, 33 C. hyointestinalis/fetus, 10 C. lari, 4 C. coli, 32 unidentified). There were 737 human and 104 pet C. jejuni/coli strains assigned to 154 and 49 STs, respectively. Dog, particularly puppy, owners were at increased risk of infection with pet-associated STs. In 2/68 cases vs. 0.134/68 expected by chance, a pet and its owner were infected with an identical ST (ST45, ST658). Although common sources of infection and directionality of transmission between pets and humans were unknown, dog ownership significantly increased the risk for pet-associated human C. jejuni/coli infection and isolation of identical strains in humans and their pets occurred significantly more often than expected.

  14. Infrared thermometry study of nanofluid pool boiling phenomena

    PubMed Central

    2011-01-01

    Infrared thermometry was used to obtain first-of-a-kind, time- and space-resolved data for pool boiling phenomena in water-based nanofluids with diamond and silica nanoparticles at low concentration (<0.1 vol.%). In addition to macroscopic parameters like the average heat transfer coefficient and critical heat flux [CHF] value, more fundamental parameters such as the bubble departure diameter and frequency, growth and wait times, and nucleation site density [NSD] were directly measured for a thin, resistively heated, indium-tin-oxide surface deposited onto a sapphire substrate. Consistent with other nanofluid studies, the nanoparticles caused deterioration in the nucleate boiling heat transfer (by as much as 50%) and an increase in the CHF (by as much as 100%). The bubble departure frequency and NSD were found to be lower in nanofluids compared with water for the same wall superheat. Furthermore, it was found that a porous layer of nanoparticles built up on the heater surface during nucleate boiling, which improved surface wettability compared with the water-boiled surfaces. Using the prevalent nucleate boiling models, it was possible to correlate this improved surface wettability to the experimentally observed reductions in the bubble departure frequency, NSD, and ultimately to the deterioration in the nucleate boiling heat transfer and the CHF enhancement. PMID:21711754

  15. Effects of commercial marinade seasoning and a natural blend of cultured sugar and vinegar on Campylobacter jejuni and Salmonella Typhimurium and the texture of chicken breasts.

    PubMed

    Park, Na Yoon; Hong, Soo Hyeon; Yoon, Ki Sun

    2014-03-01

    Marination using various ingredients has been widely used to improve microbial safety and quality of chicken products at retail markets. The objective of this study was to investigate the effects of commercial marinade seasoning and cultured sugar/vinegar blend on Campylobacter jejuni and Salmonella Typhimurium populations during refrigerated storage. In addition, their effects on the texture of precooked chicken breasts during frozen and refrigerated storage was investigated. Chicken breasts inoculated with 4.5 to 5.0 log cfu/g of C. jejuni and Salmonella Typhimurium were treated with 3% cultured sugar/vinegar blend with and without 0.6% polish rub seasoning containing 32% herb content. Breasts were then vacuum-packaged and stored at 4 and 10°C. Survival and growth curves were fitted to the Baranyi equation to determine survival and growth kinetics of C. jejuni and Salmonella Typhimurium. In addition, the vacuum-packaged precooked chicken breasts with different marination treatments were subjected to 3 freeze-thaw cycles and shear force was measured. At 4°C, the populations of C. jejuni and Salmonella Typhimurium decreased, regardless of treatment group during storage. The greatest survival for C. jejuni was observed in untreated chicken breasts. At 10°C, the growth of Salmonella Typhimurium was completely prevented in precooked chicken breasts treated with 3% cultured sugar/vinegar blend, regardless of the presence of 0.6% seasoning. The 3% cultured sugar/vinegar blend also improved the tenderness of frozen chicken breasts and refrigerated, ready-to-eat chicken breast. Therefore, a natural blend of cultured sugar and vinegar can be used as antimicrobial and texture-modifying agents for poultry meat and poultry products.

  16. Analysis of the LIV System of Campylobacter jejuni Reveals Alternative Roles for LivJ and LivK in Commensalism beyond Branched-Chain Amino Acid Transport ▿

    PubMed Central

    Ribardo, Deborah A.; Hendrixson, David R.

    2011-01-01

    Campylobacter jejuni is a leading cause of diarrheal disease in humans and an intestinal commensal in poultry and other agriculturally important animals. These zoonotic infections result in significant amounts of C. jejuni present in the food supply to contribute to disease in humans. We previously found that a transposon insertion in Cjj81176_1038, encoding a homolog of the Escherichia coli LivJ periplasmic binding protein of the leucine, isoleucine, and valine (LIV) branched-chain amino acid transport system, reduced the commensal colonization capacity of C. jejuni 81-176 in chicks. Cjj81176_1038 is the first gene of a six-gene locus that encodes homologous components of the E. coli LIV system. By analyzing mutants with in-frame deletions of individual genes or pairs of genes, we found that this system constitutes a LIV transport system in C. jejuni responsible for a high level of leucine acquisition and, to a lesser extent, isoleucine and valine acquisition. Despite each LIV protein being required for branched-chain amino acid transport, only the LivJ and LivK periplasmic binding proteins were required for wild-type levels of commensal colonization of chicks. All LIV permease and ATPase components were dispensable for in vivo growth. These results suggest that the biological functions of LivJ and LivK for colonization are more complex than previously hypothesized and extend beyond a role for binding and acquiring branched-chain amino acids during commensalism. In contrast to other studies indicating a requirement and utilization of other specific amino acids for colonization, acquisition of branched-chain amino acids does not appear to be a determinant for C. jejuni during commensalism. PMID:21949065

  17. A seventeen-year observation of the antimicrobial susceptibility of clinical Campylobacter jejuni and the molecular mechanisms of erythromycin-resistant isolates in Beijing, China.

    PubMed

    Zhou, Jiyuan; Zhang, Maojun; Yang, Wanna; Fang, Yuqing; Wang, Guiqiang; Hou, Fengqin

    2016-01-01

    To investigate the dynamic development of the antimicrobial resistance of Campylobacter jejuni isolated from human diarrhea in Beijing, China, between 1994 and 2010, and to further analyze the molecular mechanisms of erythromycin-resistant strains. Susceptibility tests were performed on 203 non-duplicate clinical C. jejuni strains against eight common antibiotics using the standard agar dilution method. The molecular determinants were further studied in the erythromycin (ERY) non-susceptible strains. The analysis focused on the 23S rRNA gene, the rplD and rplV ribosomal genes, the ermB gene, and the regulatory region of the CmeABC efflux pump. The rates of resistance of C. jejuni to ciprofloxacin (CIP), nalidixic acid (NAL), doxycycline (DOX), tetracycline (TET), florfenicol (FFC), and chloramphenicol (CHL) increased significantly over the period studied (all p<0.05). Similarly, the proportions of resistant patterns (CIP-NAL-DOX-TET, CIP-NAL-DOX-TET-FFC, and CIP-NAL-DOX-TET-CHL) increased remarkably. In this study, 4.4% (9/203) of C. jejuni strains were ERY non-susceptible. The A2075G mutation in the 23S rRNA was found in all of the resistant strains except cj8091, which harbored the ermB gene. Interestingly, the ermB gene was also detected in intermediately resistant isolates, and the earliest ermB-positive strain cj94473 was derived in 1994. Moreover, none of the ribosomal rplD or rplV genes harbored mutations that have been described to confer resistance to macrolides. Different mutations affecting the regulatory region of the CmeABC efflux pump were also found. This is the first comprehensive study on the recent trend in antimicrobial resistance and the molecular mechanisms of macrolide resistance in clinical C. jejuni strains isolated in China. More stringent monitoring and regulation of human and animal antimicrobial use are warranted. Copyright © 2015 The Authors. Published by Elsevier Ltd.. All rights reserved.

  18. Theoretical modeling of CHF for near-saturated pool boiling and flow boiling from short heaters using the interfacial lift-off criterion

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Mudawar, I.; Galloway, J.E.; Gersey, C.O.

    Pool boiling and flow boiling were examined for near-saturated bulk conditions in order to determine the critical heat flux (CHF) trigger mechanism for each. Photographic studies of the wall region revealed features common to both situations. At fluxes below CHF, the vapor coalesces into a wavy layer which permits wetting only in wetting fronts, the portions of the liquid-vapor interface which contact the wall as a result of the interfacial waviness. Close examination of the interfacial features revealed the waves are generated from the lower edge of the heater in pool boiling and the heater`s upstream region in flow boiling.more » Wavelengths follow predictions based upon the Kelvin-Helmholtz instability criterion. Critical heat flux in both cases occurs when the pressure force exerted upon the interface due to interfacial curvature, which tends to preserve interfacial contact with the wall prior to CHF, is overcome by the momentum of vapor at the site of the first wetting front, causing the interface to lift away from the wall. It is shown this interfacial lift-off criterion facilitates accurate theoretical modeling of CHF in pool boiling and in flow boiling in both straight and curved channels.« less

  19. A Fundamental Study of Nucleate Pool Boiling Under Microgravity

    NASA Technical Reports Server (NTRS)

    Ervin, Jamie S.; Merte, Herman, Jr.

    1996-01-01

    An experimental study of incipient boiling in short-term microgravity and with a/g = +/- 1 for pool boiling was performed. Calibrated thin gold films sputtered on a smoothly polished quartz surface were used simultaneously for thermal-resistance measurements and heating of the boiling surface. The gold films were used for both transient and quasi-steady heating surface temperature measurements. Two test vessels were constructed for precise measurement and control of fluid temperature and pressure: a laboratory pool boiling vessel for the a/g = +/- 1 experiments and a pool boiling vessel designed for the 131 m free-fall in the NASA Lewis Research Center Microgravity Research Facility for the microgravity tests. Measurements included the heater surface temperature, the pressure near the heating surface, the bulk liquid temperatures. High speed photography (up to 1,000 frames per second) was used in the experiments. With high quality microgravity and the measured initial temperature of the quiescent test fluid, R113, the temperature distribution in the liquid at the moment of boiling inception resulting from an imposed step in heat flux is known with a certainty not possible previously. The types of boiling propagation across the large flat heating surface, some observed here for the first time, are categorized; the conditions necessary for their occurrence are described. Explosive boiling propagation with a striking pattern of small scale protuberances over the entire vapor mass periphery not observed previously at low heat flux levels (on the order of 5 W/cm(exp 2)) is described. For the heater surface with a/g = -1, a step in the heater surface temperature of short duration was imposed. The resulting liquid temperature distribution at the moment of boiling inception was different from that obtained with a step in heat flux.

  20. Heat Transfer Performances of Pool Boiling on Metal-Graphite Composite Surfaces

    NASA Technical Reports Server (NTRS)

    Zhang, Nengli; Chao, David F.; Yang, Wen-Jei

    2000-01-01

    Nucleate boiling, especially near the critical heat flux (CHF), can provide excellent economy along with high efficiency of heat transfer. However, the performance of nucleate boiling may deteriorate in a reduced gravity environment and the nucleate boiling usually has a potentially dangerous characteristic in CHF regime. That is, any slight overload can result in burnout of the boiling surface because the heat transfer will suddenly move into the film-boiling regime. Therefore, enhancement of nucleate boiling heat transfer becomes more important in reduced gravity environments. Enhancing nucleate boiling and critical heat flux can be reached using micro-configured metal-graphite composites as the boiling surface. Thermocapillary force induced by temperature difference between the graphite-fiber tips and the metal matrix, which is independent of gravity, will play an important role in bubble detachment. Thus boiling heat transfer performance does not deteriorate in a reduced-gravity environment. Based on the existing experimental data, and a two-tier theoretical model, correlation formulas are derived for nucleate boiling on the copper-graphite and aluminum-graphite composite surfaces, in both the isolated and coalesced bubble regimes. Experimental studies were performed on nucleate pool boiling of pentane on cooper-graphite (Cu-Gr) and aluminum-graphite (Al-Gr) composite surfaces with various fiber volume concentrations for heat fluxes up to 35 W per square centimeter. It is revealed that a significant enhancement in boiling heat transfer performance on the composite surfaces is achieved, due to the presence of micro-graphite fibers embedded in the matrix. The onset of nucleate boiling (the isolated bubble regime) occurs at wall superheat of about 10 C for the Cu-Gr surface and 15 C for the Al-Gr surface, much lower than their respective pure metal surfaces. Transition from an isolated bubble regime to a coalesced bubble regime in boiling occurs at a superheat of

  1. Evaluation and single-laboratory verification of a proposed modification to the U.S. Food and Drug Administration method for detection and identification of Campylobacter jejuni or Campylobacter coli from raw silo milk.

    PubMed

    Gharst, Greg; Bark, Don H; Newkirk, Robert; Guillen, Lacey; Wang, Qian; Abeyta, Carlos

    2013-01-01

    The current U.S. Food and Drug Administration (FDA) methodology for detection of Campylobacter, a leading source for foodborne illness, is outdated. The purpose of this study, therefore, was to improve and update the cultural and identification methods found in the FDA/Bacteriological Analytical Manual (BAM). Raw silo milk samples containing typical and atypical strains of Campylobacter jejuni and Campylobacter coli at different levels (5 CFU/25 g, 50 CFU/25 g, and 125 CFU/25 g) were analyzed. Valid results were obtained from 240 test portions. Six inoculated (at the levels described above) and two uninoculated samples were sent to a participating laboratory to mimic a "real-world" scenario. These combined data indicated that the use of sheep blood in combination with enrichment is not necessary. R & F Campylobacter jejuni/Campylobacter coli Chromogenic Plating Medium is significantly (P < 0.05) more sensitive for detection of C. jejuni or C. coli at low inoculation levels than the modified Cefoperazone Charcoal Deoxycholate Agar used in the BAM. The quantitative PCR method described demonstrated rapid confirmation and identification of C. jejuni or C. coli. It reduced the time to isolate C. jejuni or C. coli, and increased the sensitivity compared to the current BAM protocol.

  2. Development of Flow Boiling and Condensation Experiment on the International Space Station- Normal and Low Gravity Flow Boiling Experiment Development and Test Results

    NASA Technical Reports Server (NTRS)

    Nahra, Henry K.; Hall, Nancy R.; Hasan, Mohammad M.; Wagner, James D.; May, Rochelle L.; Mackey, Jeffrey R.; Kolacz, John S.; Butcher, Robert L.; Frankenfield, Bruce J.; Mudawar, Issam; hide

    2013-01-01

    Flow boiling and condensation have been identified as two key mechanisms for heat transport that are vital for achieving weight and volume reduction as well as performance enhancement in future space systems. Since inertia driven flows are demanding on power usage, lower flows are desirable. However, in microgravity, lower flows are dominated by forces other than inertia (like the capillary force). It is of paramount interest to investigate limits of low flows beyond which the flow is inertial enough to be gravity independent. One of the objectives of the Flow Boiling and Condensation Flight Experiment sets to investigate these limits for flow boiling and condensation. A two-phase flow loop consisting of a Flow Boiling Module and two Condensation Modules has been developed to experimentally study flow boiling condensation heat transfer in the reduced gravity environment provided by the reduced gravity platform. This effort supports the development of a flow boiling and condensation facility for the International Space Station (ISS). The closed loop test facility is designed to deliver the test fluid, FC-72 to the inlet of any one of the test modules at specified thermodynamic and flow conditions. The zero-g-aircraft tests will provide subcooled and saturated flow boiling critical heat flux and flow condensation heat transfer data over wide range of flow velocities. Additionally, these tests will verify the performance of all gravity sensitive components, such as evaporator, condenser and accumulator associated with the two-phase flow loop. We will present in this paper the breadboard development and testing results which consist of detailed performance evaluation of the heater and condenser combination in reduced and normal gravity. We will also present the design of the reduced gravity aircraft rack and the results of the ground flow boiling heat transfer testing performed with the Flow Boiling Module that is designed to investigate flow boiling heat transfer and

  3. Cryogenic Boil-Off Reduction System Testing

    NASA Technical Reports Server (NTRS)

    Plachta, David W.; Johnson, Wesley L.; Feller, Jeffrey R.

    2014-01-01

    Cryogenic propellants such as liquid hydrogen (LH2) and liquid oxygen (LO2) are a part of NASA's future space exploration due to the high specific impulse that can be achieved using engines suitable for moving 10's to 100's of metric tons of payload mass to destinations outside of low earth orbit. However, the low storage temperatures of LH2 and LO2 cause substantial boil-off losses for missions with durations greater than several days. The losses can be greatly reduced by incorporating high performance cryocooler technology to intercept heat load to the propellant tanks and by the integration of self-supporting multi-layer insulation. The active thermal control technology under development is the integration of the reverse turbo- Brayton cycle cryocooler to the propellant tank through a distributed cooling network of tubes coupled to a shield in the tank insulation and to the tank wall itself. Also, the self-supporting insulation technology was utilized under the shield to obtain needed tank applied LH2 performance. These elements were recently tested at NASA Glenn Research Center in a series of three tests, two that reduced LH2 boil-off and one to eliminate LO2 boil-off. This test series was conducted in a vacuum chamber that replicated the vacuum of space and the temperatures of low Earth orbit. The test results show that LH2 boil-off was reduced 60% by the cryocooler system operating at 90K and that robust LO2 zero boil-off storage, including full tank pressure control was achieved.

  4. Prevalence of three campylobacter species, C. jejuni, C. coli, and C. lari, using multilocus sequence typing in wild birds of the Mid-Atlantic region, USA.

    PubMed

    Keller, Judith I; Shriver, W Gregory

    2014-01-01

    Campylobacter jejuni is responsible for the majority of bacterial foodborne gastroenteritis in the US, usually due to the consumption of undercooked poultry. Research on which avian species transmit the bacterium is limited, especially in the US. We sampled wild birds in three families-Anatidae, Scolopacidae, and Laridae-in eastern North America to determine the prevalence and specific strains of Campylobacter. The overall prevalence of Campylobacter spp. was 9.2% for all wild birds sampled (n = 781). Campylobacter jejuni was the most prevalent species (8.1%), while Campylobacter coli and Campylobacter lari prevalence estimates were low (1.4% and 0.3%, respectively). We used multilocus sequence typing PCR specific to C. jejuni to characterize clonal complexes and sequence types isolated from wild bird samples and detected 13 novel sequence types, along with a clonal complex previously only associated with human disease (ST-658). Wild birds share an increasing amount of habitat with humans as more landscapes become fragmented and developed for human needs. Wild birds are and will remain an important aspect of public health due to their ability to carry and disperse emerging zoonotic pathogens or their arthropod vectors. As basic information such as prevalence is limited or lacking from a majority of wild birds in the US, this study provides further insight into Campylobacter epidemiology, host preference, and strain characterization of C. jejuni.

  5. Feasibility study of a brine boiling machine by solar energy

    NASA Astrophysics Data System (ADS)

    Phayom, W.

    2018-06-01

    This study presented the technical and operational feasibility of brine boiling machine by using solar energy instead of firewood or husk for salt production. The solar salt brine boiling machine consisted of a boiling chamber with an enhanced thermal efficiency through use of a solar brine heater. The stainless steel solar salt brine boiling chamber had dimensions of 60 cm x 70 cm x 20 cm. The steel brine heater had dimensions of 70 cm x 80 cm x 20 cm. The tilt angle of both the boiling chamber and brine heater was 20 degrees from horizontal. The brine temperature in the reservoir tank was 42°C with a flow rate of 6.64 L/h discharging into the solar boiling machine. It was found that the thermal efficiency and overall efficiency of the solar salt brine boiling machine were 0.63 and 0.38, respectively at a solar irradiance of 787.6 W/m2. The results shows that the potential of using solar energy for salt production system is feasible.

  6. Skin whitening and anti-corrugation activities of glycoprotein fractions from liquid extracts of boiled sea cucumber.

    PubMed

    Kim, So Jung; Park, So Yun; Hong, Sun-Mee; Kwon, Eun-Hye; Lee, Taek-Kyun

    2016-10-01

    To determine skin whitening and wrinkle improvement efficacy, glycoprotein fractions were extracted from liquid extracts of boiled sea cucumber and their effects on tyrosine and elastase inhibitory activities were assayed. Fractions above and below 50 kDa (>50 kDa and <50 kDa) were extracted via a series of steps involving: boiling, filtering, desalting and freeze drying. Cytotoxicity, skin whitening and wrinkle-removing effects of boiled liquid were determined. Our MTT data showed that neither glycoprotein fraction of boiled liquid induces cellular cytotoxicity up to a concentration of 10 mg/mL treatment of the mouse melanoma cell line, B16F10, with 10 mg/mL >50 kDa enhanced tyrosinase and elastase inhibitory activities by 50.84% and 28.78%, respectively. Correlations of the >50 kDa concentration with tyrosinase inhibitory (R2 = 0.968) and elastase inhibitory (R2 = 0.983) efficacy were significant. >50 kDa glycoprotein fraction isolated from liquid extracts of boiled sea cucumber, which can serve as a functional cosmetic ingredient for whitening and wrinkle improvement of skin. Copyright © 2016 Hainan Medical University. Production and hosting by Elsevier B.V. All rights reserved.

  7. Liquid metal boiling inception

    NASA Technical Reports Server (NTRS)

    Sabin, C. M.; Poppendiek, H. F.; Mouritzen, G.; Meckel, P. T.; Cloakey, J. E.

    1972-01-01

    An experimental study of the inception of boiling in potassium in forced convection is reported. The boiler consisted of a 0.19-inch inside diameter, niobium-1% zirconium boiler tube approximately six feet long. Heating was accomplished by direct electrical tube wall conduction. Experiments were performed with both all-liquid fill and two-phase fill startup sequences and with a range of flow rates, saturation temperatures, inert gas levels, and fill liquid temperatures. Superheat of the liquid above the equilibrium saturation temperature was observed in all the experiments. Incipient boiling liquid superheat ranged from a few degrees to several hundred. Comparisons of these data with other data and with several analytical treatments are presented.

  8. Critical role of LuxS in the virulence of Campylobacter jejuni in a guinea pig model of abortion.

    PubMed

    Plummer, Paul; Sahin, Orhan; Burrough, Eric; Sippy, Rachel; Mou, Kathy; Rabenold, Jessica; Yaeger, Mike; Zhang, Qijing

    2012-02-01

    Previous studies on Campylobacter jejuni have demonstrated the role of LuxS in motility, cytolethal distending toxin production, agglutination, and intestinal colonization; however, its direct involvement in virulence has not been reported. In this study, we demonstrate a direct role of luxS in the virulence of C. jejuni in two different animal hosts. The IA3902 strain, a highly virulent sheep abortion strain recently described by our laboratory, along with its isogenic luxS mutant and luxS complement strains, was inoculated by the oral route into both a pregnant guinea pig virulence model and a chicken colonization model. In both cases, the IA3902 luxS mutant demonstrated a complete loss of ability to colonize the intestinal tract. In the pregnant model, the mutant also failed to induce abortion, while the wild-type strain was highly abortifacient. Genetic complementation of the luxS gene fully restored the virulent phenotype in both models. Interestingly, when the organism was inoculated into guinea pigs by the intraperitoneal route, no difference in virulence (abortion induction) was observed between the luxS mutant and the wild-type strain, suggesting that the defect in virulence following oral inoculation is likely associated with a defect in colonization and/or translocation of the organism out of the intestine. These studies provide the first direct evidence that LuxS plays an important role in the virulence of C. jejuni using an in vivo model of natural disease.

  9. Prevalence of Campylobacter jejuni and Campylobacter coli in captive wildlife species of India

    PubMed Central

    Prince Milton, A. A.; Agarwal, R. K.; Priya, G. B.; Saminathan, M.; Aravind, M.; Reddy, A.; Athira, C. K.; Anjay; Ramees, T. P.; Dhama, K.; Sharma, A. K.; Kumar, A.

    2017-01-01

    Campylobacteriosis is an important zoonotic disease and the prevalence of Campylobacter is largely unknown in the wildlife of India. A total of 370 samples, comprising of 314 fresh faecal samples from apparently healthy captive wild animals and birds, 30 stool swabs from animal care takers and 26 samples of the animals’ food and water were collected from G. B. Pant High Altitude Zoo, Nainital, Kanpur Zoo, Wildlife Park, IVRI and the Post Graduate Research Institute in Animal Sciences (PGRIAS), Chennai, Tamilnadu from August 2014 to May 2015. Samples were processed for cultural isolation, direct PCR and multiplex PCR for species confirmation. To decipher the genetic diversity, the 16S rRNA gene was amplified, sequenced and analyzed. Based on isolation, the overall occurrence rate of Campylobacter spp. was 0.8% (3/370), being 2.94% (3/102) for captive wild birds. Three Campylobacter jejuni were isolated from silver pheasants, lady amherest pheasants and saras cranes. Direct PCR assay showed the overall occurrence rate of Campylobacter spp. to be 4.77% (15/315), being 1.58% (2/126) for captive wild ruminants, 5.81% (5/86) for non-ruminants and 7.84% (8/102) for birds. All the isolates were identified as C. jejuni. PMID:29163646

  10. ISOLATION AND MOLECULAR IDENTIFICATION OF POTENTIALLY PATHOGENIC Escherichia coli AND Campylobacter jejuni IN FERAL PIGEONS FROM AN URBAN AREA IN THE CITY OF LIMA, PERU

    PubMed Central

    CABALLERO, Moisés; RIVERA, Isabel; JARA, Luis M.; ULLOA-STANOJLOVIC, Francisco M.; SHIVA, Carlos

    2015-01-01

    SUMMARY Feral pigeons (Columbia livia) live in close contact with humans and other animals. They can transmit potentially pathogenic and zoonotic agents. The objective of this study was to isolate and detect strains of diarrheagenic Escherichia coli and Campylobacter jejuni of urban feral pigeons from an area of Lima, Peru. Fresh dropping samples from urban parks were collected for microbiological isolation of E. coli strains in selective agar, and Campylobacter by filtration method. Molecular identification of diarrheagenic pathotypes of E.coli and Campylobacter jejuni was performed by PCR. Twenty-two parks were sampled and 16 colonies of Campylobacter spp. were isolated. The 100% of isolates were identified as Campylobacter jejuni. Furthermore, 102 colonies of E. coliwere isolated and the 5.88% resulted as Enteropathogenic (EPEC) type and 0.98% as Shiga toxin-producing E. coli (STEC). The urban feral pigeons of Lima in Peru can act as a reservoir or carriers of zoonotic potentially pathogenic enteric agents. PMID:26603225

  11. Influence of Asellus aquaticus on Escherichia coli, Klebsiella pneumoniae, Campylobacter jejuni and naturally occurring heterotrophic bacteria in drinking water.

    PubMed

    Christensen, Sarah C B; Nissen, Erling; Arvin, Erik; Albrechtsen, Hans-Jørgen

    2012-10-15

    Water lice, Asellus aquaticus (isopoda), frequently occur in drinking water distribution systems where they are a nuisance to consumers and water utilities. Whether they are solely an aesthetic problem or also affect the microbial water quality is a matter of interest. We studied the influence of A. aquaticus on microbial water quality in non-chlorinated drinking water in controlled laboratory experiments. Pure cultures of the indicator organisms Escherichia coli and Klebsiella pneumoniae and the pathogen Campylobacter jejuni as well as naturally occurring heterotrophic drinking water bacteria (measured as heterotrophic plate counts, HPC) were investigated in microcosms at 7 °C, containing non-sterilised drinking water, drinking water sediment and A. aquaticus collected from a non-chlorinated ground water based drinking water supply system. Concentrations of E. coli, K. pneumoniae and C. jejuni decreased over time, following a first order decay with half lives of 5.3, 18.4 and 1.3 days, respectively. A. aquaticus did not affect survival of indicators and pathogens substantially whereas HPC were influenced by presence of dead A. aquaticus. Growth rates increased with an average of 48% for bacteria grown on R-2A agar and an average of 83% for bacteria grown on yeast extract agar when dead A. aquaticus were present compared to no and living A. aquaticus present. A. aquaticus associated E. coli, K. pneumoniae and C. jejuni were measured (up to 25 per living and 500 per dead A. aquaticus) and so were A. aquaticus associated heterotrophic bacteria (>1.8*10(4) CFU per living and >6*10(4) CFU per dead A. aquaticus). A. aquaticus did not serve as an optimised habitat that increased survival of indicators and pathogens, since A. aquaticus associated E. coli, K. pneumoniae and C. jejuni were only measured as long as the bacteria were also present in the water and sediment. Copyright © 2012 Elsevier Ltd. All rights reserved.

  12. An investigation of transition boiling mechanisms of subcooled water under forced convective conditions

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Kwang-Won, Lee; Sang-Yong, Lee

    1995-09-01

    A mechanistic model for forced convective transition boiling has been developed to investigate transition boiling mechanisms and to predict transition boiling heat flux realistically. This model is based on a postulated multi-stage boiling process occurring during the passage time of the elongated vapor blanket specified at a critical heat flux (CHF) condition. Between the departure from nucleate boiling (DNB) and the departure from film boiling (DFB) points, the boiling heat transfer is established through three boiling stages, namely, the macrolayer evaporation and dryout governed by nucleate boiling in a thin liquid film and the unstable film boiling characterized by themore » frequent touches of the interface and the heated wall. The total heat transfer rates after the DNB is weighted by the time fractions of each stage, which are defined as the ratio of each stage duration to the vapor blanket passage time. The model predictions are compared with some available experimental transition boiling data. The parametric effects of pressure, mass flux, inlet subcooling on the transition boiling heat transfer are also investigated. From these comparisons, it can be seen that this model can identify the crucial mechanisms of forced convective transition boiling, and that the transition boiling heat fluxes including the maximum heat flux and the minimum film boiling heat flux are well predicted at low qualities/high pressures near 10 bar. In future, this model will be improved in the unstable film boiling stage and generalized for high quality and low pressure situations.« less

  13. Transient nucleate pool boiling in microgravity: Some initial results

    NASA Technical Reports Server (NTRS)

    Merte, Herman, Jr.; Lee, H. S.; Ervin, J. S.

    1994-01-01

    Variable gravity provides an opportunity to test the understanding of phenomena which are considered to depend on buoyancy, such as nucleate pool boiling. The active fundamental research in nucleate boiling has sought to determine the mechanisms or physical processes responsible for its high effectiveness, manifested by the high heat flux levels possible with relatively low temperature differences. Earlier research on nucleate pool boiling at high gravity levels under steady conditions demonstrated quantitatively that the heat transfer is degraded as the buoyancy normal to the heater surfaced increases. Correspondingly, it was later shown, qualitatively for short periods of time only, that nucleate boiling heat transfer is enhanced as the buoyancy normal to the heater surface is reduced. It can be deduced that nucleate pool boiling can be sustained as a quasi-steady process provided that some means is available to remove the vapor generated from the immediate vicinity of the heater surface. One of the objectives of the research, the initial results of which are presented here, is to quantify the heat transfer associated with boiling in microgravity. Some quantitative results of nucleate pool boiling in high quality microgravity (a/g approximately 10(exp -5)) of 5s duration, obtained in an evacuated drop tower, are presented here. These experiments were conducted as precursors of longer term space experiments. A transient heating technique is used, in which the heater surface is a transparent gold film sputtered on a qua rtz substrate, simultaneously providing the mean surface temperature from resistance thermometry and viewing of the boiling process both from beneath and across the surface. The measurement of the transient mean heater surface temperature permits the computation, by numerical means, of the transient mean heat transfer coefficient. The preliminary data obtained demonstrates that a quasi-steady boiling process can occur in microgravity if the bulk

  14. Electrically Driven Liquid Film Boiling Experiment

    NASA Technical Reports Server (NTRS)

    Didion, Jeffrey R.

    2016-01-01

    This presentation presents the science background and ground based results that form the basis of the Electrically Driven Liquid Film Boiling Experiment. This is an ISS experiment that is manifested for 2021. Objective: Characterize the effects of gravity on the interaction of electric and flow fields in the presence of phase change specifically pertaining to: a) The effects of microgravity on the electrically generated two-phase flow. b) The effects of microgravity on electrically driven liquid film boiling (includes extreme heat fluxes). Electro-wetting of the boiling section will repel the bubbles away from the heated surface in microgravity environment. Relevance/Impact: Provides phenomenological foundation for the development of electric field based two-phase thermal management systems leveraging EHD, permitting optimization of heat transfer surface area to volume ratios as well as achievement of high heat transfer coefficients thus resulting in system mass and volume savings. EHD replaces buoyancy or flow driven bubble removal from heated surface. Development Approach: Conduct preliminary experiments in low gravity and ground-based facilities to refine technique and obtain preliminary data for model development. ISS environment required to characterize electro-wetting effect on nucleate boiling and CHF in the absence of gravity. Will operate in the FIR - designed for autonomous operation.

  15. SUPERHEATING IN A BOILING WATER REACTOR

    DOEpatents

    Treshow, M.

    1960-05-31

    A boiling-water reactor is described in which the steam developed in the reactor is superheated in the reactor. This is accomplished by providing means for separating the steam from the water and passing the steam over a surface of the fissionable material which is not in contact with the water. Specifically water is boiled on the outside of tubular fuel elements and the steam is superheated on the inside of the fuel elements.

  16. Genetic basis and clonal population structure of antibiotic resistance in Campylobacter jejuni isolated from broiler carcasses in Belgium

    USDA-ARS?s Scientific Manuscript database

    The aim of this study was to investigate the population structure and antimicrobial resistance profiles of a set of Campylobacter jejuni strains isolated from broiler carcasses in Belgium, and to further analyze the molecular mechanisms responsible for the resistance phenotypes. Minimum inhibitory c...

  17. Criticality in the slowed-down boiling crisis at zero gravity.

    PubMed

    Charignon, T; Lloveras, P; Chatain, D; Truskinovsky, L; Vives, E; Beysens, D; Nikolayev, V S

    2015-05-01

    Boiling crisis is a transition between nucleate and film boiling. It occurs at a threshold value of the heat flux from the heater called CHF (critical heat flux). Usually, boiling crisis studies are hindered by the high CHF and short transition duration (below 1 ms). Here we report on experiments in hydrogen near its liquid-vapor critical point, in which the CHF is low and the dynamics slow enough to be resolved. As under such conditions the surface tension is very small, the experiments are carried out in the reduced gravity to preserve the conventional bubble geometry. Weightlessness is created artificially in two-phase hydrogen by compensating gravity with magnetic forces. We were able to reveal the fractal structure of the contour of the percolating cluster of the dry areas at the heater that precedes the boiling crisis. We provide a direct statistical analysis of dry spot areas that confirms the boiling crisis at zero gravity as a scale-free phenomenon. It was observed that, in agreement with theoretical predictions, saturated boiling CHF tends to zero (within the precision of our thermal control system) in zero gravity, which suggests that the boiling crisis may be observed at any heat flux provided the experiment lasts long enough.

  18. The influence of feeding crimped kernel maize silage on growth performance and intestinal colonization with Campylobacter jejuni of broilers.

    PubMed

    Ranjitkar, Samir; Engberg, Ricarda Margarete

    2016-01-01

    An infection trial and a production trial over 35 days were conducted in parallel to study the influence of feeding crimped kernel maize silage (CKMS) on the intestinal Campylobacter jejuni colonization and broiler performance, respectively. The CKMS was used at dietary inclusion levels of 15% and 30% in maize-based diets. Broilers were orally inoculated with 2 × 10(5) log cfu/ml C. jejuni on day 14. Four birds from each pen were randomly selected and killed by cervical dislocation on days 3, 6, 9, 14 and 21 post infection and intestinal contents from ileum, caeca and rectum as well as liver samples were taken. Body weight and feed consumption of broilers were registered on days 13, 22 and 35. On day 35, litter dry matter (DM) was measured and the condition of the foot pads was evaluated. There was no significant effect of CKMS on the colonization of C. jejuni. Body weight of the broilers supplemented with 15% CKMS was comparable with the control maize-based feed, whereas addition of 30% CKMS reduced broiler body weight (P < 0.001). However, DM intake and feed conversion ratio were the same in all three dietary treatments. Furthermore, the foot pad condition of broilers significantly improved with the inclusion of CKMS on broiler diets as a result of a higher DM content in the litter material. It is concluded that CKMS did not influence intestinal Campylobacter colonization, but improved the foot pad health of broilers.

  19. The initiation of boiling during pressure transients. [water boiling on metal surfaces

    NASA Technical Reports Server (NTRS)

    Weisman, J.; Bussell, G.; Jashnani, I. L.; Hsieh, T.

    1973-01-01

    The initiation of boiling of water on metal surfaces during pressure transients has been investigated. The data were obtained by a new technique in which light beam fluctuations and a pressure signal were simultaneously recorded on a dual beam oscilloscope. The results obtained agreed with those obtained using high speed photography. It was found that, for water temperatures between 90-150 C, the wall superheat required to initiate boiling during a rapid pressure transient was significantly higher than required when the pressure was slowly reduced. This result is explained by assuming that a finite time is necessary for vapor to fill the cavity at which the bubble originates. Experimental measurements of this time are in reasonably good agreement with calculations based on the proposed theory. The theory includes a new procedure for estimating the coefficient of vaporization.

  20. Generation of the membrane potential and its impact on the motility, ATP production and growth in Campylobacter jejuni

    USDA-ARS?s Scientific Manuscript database

    The generation of an electrical membrane potential (''), the major constituent of the proton motive force (pmf) is crucial for the ATP synthesis, bacterial growth and motility. The pmf drives the rotation of flagella and is vital for the microaerophilic human pathogen Campylobacter jejuni to coloniz...

  1. 40 CFR 180.1056 - Boiled linseed oil; exemption from requirement of tolerance.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... “boiled linseed oil.” This exemption is limited to use on rice before edible parts form. [46 FR 33270... 40 Protection of Environment 23 2010-07-01 2010-07-01 false Boiled linseed oil; exemption from... From Tolerances § 180.1056 Boiled linseed oil; exemption from requirement of tolerance. Boiled linseed...

  2. Faecal contamination of a municipal drinking water distribution system in association with Campylobacter jejuni infections.

    PubMed

    Pitkänen, Tarja; Miettinen, Ilkka T; Nakari, Ulla-Maija; Takkinen, Johanna; Nieminen, Kalle; Siitonen, Anja; Kuusi, Markku; Holopainen, Arja; Hänninen, Marja-Liisa

    2008-09-01

    After heavy rains Campylobacter jejuni together with high counts of Escherichia coli, other coliforms and intestinal enterococci were detected from drinking water of a municipal distribution system in eastern Finland in August 2004. Three patients with a positive C. jejuni finding, who had drunk the contaminated water, were identified and interviewed. The pulsed-field gel electrophoresis (PFGE) genotypes from the patient samples were identical to some of the genotypes isolated from the water of the suspected contamination source. In addition, repetitive DNA element analysis (rep-PCR) revealed identical patterns of E. coli and other coliform isolates along the distribution line. Further on-site technical investigations revealed that one of the two rainwater gutters on the roof of the water storage tower had been in an incorrect position and rainwater had flushed a large amount of faecal material from wild birds into the drinking water. The findings required close co-operation between civil authorities, and application of cultivation and genotyping techniques strongly suggested that the municipal drinking water was the source of the infections. The faecal contamination associated with failures in cleaning and technical management stress the importance of instructions for waterworks personnel to perform maintenance work properly.

  3. NUCLEAR SUPERHEATER FOR BOILING WATER REACTOR

    DOEpatents

    Holl, R.J.; Klecker, R.W.; Graham, C.B.

    1962-05-15

    A description is given of a boiling water reactor having a superheating region integral with the core. The core consists essentially of an annular boiling region surrounding an inner superheating region. Both regions contain fuel elements and are separated by a cylindrical wall, perforations being provided in the lower portion of the cylindrical wall to permit circulation of a common water moderator between the two regions. The superheater region comprises a plurality of tubular fuel assemblies through which the steam emanating from the boiling region passes to the steam outlet. Each superheater fuel assembly has an outer double-walled cylinder, the double walls being concentrically spaced and connected together at their upper ends but open at the bottom to provide for differential thermal expansion of the inner and outer walls. Gas is entrapped in the annulus between the walls which acts as an insulating space between the fissionable material inside and the moderator outside. (AEC)

  4. Physical quality of Simental Ongole crossbred silverside meat at various boiling times

    NASA Astrophysics Data System (ADS)

    Riyanto, J.; Cahyadi, M.; Guntari, W. S.

    2018-03-01

    This study aims to determine the physical quality of silverside beef meat at various boiling times. Samples that have been used are the back thigh or silverside meat. Treatment of boiling meat included TR (meat without boiled), R15 (boiled 15 minutes), and R30 (boiled for 30 minutes). The experimental design using Completely Randomized Design with 3 replications. Each replication was done in triple physical quality test. Determination of physical quality was performed at the Livestock Industry and Processing Laboratory at Sebelas Maret University Surakarta and the Meat Technology Laboratory at the Faculty of Animal Husbandry of Gadjah Mada University. The result of variance analysis showed that boiling affect cooking loss (P≥0.05) and but did not affect (P≤0,05) pH, water holding capacity and meat tenderness. The conclusions of the study showed that boiling for 15 minutes and 30 minutes decreased the cooking loss of Simental Ongole Crossbred silverside meat. Meat physical quality of pH, water holding capacity and the value of tenderness is not affected by boiling for 15 and 30 minutes.

  5. Mosaic Structure of a Multiple-Drug-Resistant, Conjugative Plasmid from Campylobacter jejuni

    DTIC Science & Technology

    2005-01-30

    allele of each gene in the respective clones. There were three genes predicted to encode alleles of strep- tomycin-inactivating enzymes from Enterococcus ...aminoglycoside 6-adenyltransferase/E. faecium /NP_863159 3 cpp50 2599–2811 26.3 473 70 100/100 (70) Unknown of pTet plasmid/C. jejuni strain 81-176/YP_063493 4... faecium /NP_863159 24 sat4 17692–18222 37.7 180 176 94/94 (176) Streptothricin acetyltransferase/E. faecium /AAM77897 25 aphA-3 18315–19109 44.9 264

  6. Nucleate Boiling Heat Transfer Studied Under Reduced-Gravity Conditions

    NASA Technical Reports Server (NTRS)

    Chao, David F.; Hasan, Mohammad M.

    2000-01-01

    Boiling is known to be a very efficient mode of heat transfer, and as such, it is employed in component cooling and in various energy-conversion systems. In space, boiling heat transfer may be used in thermal management, fluid handling and control, power systems, and on-orbit storage and supply systems for cryogenic propellants and life-support fluids. Recent interest in the exploration of Mars and other planets and in the concept of in situ resource utilization on the Martian and Lunar surfaces highlights the need to understand how gravity levels varying from the Earth's gravity to microgravity (1g = or > g/g(sub e) = or > 10(exp -6)g) affect boiling heat transfer. Because of the complex nature of the boiling process, no generalized prediction or procedure has been developed to describe the boiling heat transfer coefficient, particularly at reduced gravity levels. Recently, Professor Vijay K. Dhir of the University of California at Los Angeles proposed a novel building-block approach to investigate the boiling phenomena in low-gravity to microgravity environments. This approach experimentally investigates the complete process of bubble inception, growth, and departure for single bubbles formed at a well-defined and controllable nucleation site. Principal investigator Professor Vijay K. Dhir, with support from researchers from the NASA Glenn Research Center at Lewis Field, is performing a series of pool boiling experiments in the low-gravity environments of the KC 135 microgravity aircraft s parabolic flight to investigate the inception, growth, departure, and merger of bubbles from single- and multiple-nucleation sites as a function of the wall superheat and the liquid subcooling. Silicon wafers with single and multiple cavities of known characteristics are being used as test surfaces. Water and PF5060 (an inert liquid) were chosen as test liquids so that the role of surface wettability and the magnitude of the effect of interfacial tension on boiling in reduced

  7. CONTINUOUS ANALYZER UTILIZING BOILING POINT DETERMINATION

    DOEpatents

    Pappas, W.S.

    1963-03-19

    A device is designed for continuously determining the boiling point of a mixture of liquids. The device comprises a distillation chamber for boiling a liquid; outlet conduit means for maintaining the liquid contents of said chamber at a constant level; a reflux condenser mounted above said distillation chamber; means for continuously introducing an incoming liquid sample into said reflux condenser and into intimate contact with vapors refluxing within said condenser; and means for measuring the temperature of the liquid flowing through said distillation chamber. (AEC)

  8. Using MOMP typing as an epidemiological tool to investigate outbreaks caused by milkborne Campylobacter jejuni isolates in California

    USDA-ARS?s Scientific Manuscript database

    We describe using major outer membrane protein (MOMP) typing as a screen to compare the C. jejuni porA gene sequences of clinical outbreak strains from human stool with the porA sequences of dairy farm strains isolated during two milkborne campylobacteriosis outbreak investigations in California. Th...

  9. A Mutator Phenotype Promoting the Emergence of Spontaneous Oxidative Stress-Resistant Mutants in Campylobacter jejuni.

    PubMed

    Dai, Lei; Sahin, Orhan; Tang, Yizhi; Zhang, Qijing

    2017-12-15

    Campylobacter jejuni is a leading cause of foodborne illnesses worldwide. As a microaerophilic organism, C. jejuni must be able to defend against oxidative stress encountered both in the host and in the environment. How Campylobacter utilizes a mutation-based mechanism for adaptation to oxidative stress is still unknown. Here we present a previously undescribed phenotypic and genetic mechanism that promotes the emergence of oxidative stress-resistant mutants. Specifically, we showed that a naturally occurring mutator phenotype, resulting from a loss of function mutation in the DNA repair enzyme MutY, increased oxidative stress resistance (OX R ) in C. jejuni We further demonstrated that MutY malfunction did not directly contribute to the OX R phenotype but increased the spontaneous mutation rate in the peroxide regulator gene perR , which functions as a repressor for multiple genes involved in oxidative stress resistance. Mutations in PerR resulted in loss of its DNA binding function and derepression of PerR-controlled oxidative stress defense genes, thereby conferring an OX R phenotype and facilitating Campylobacter survival under oxidative stress. These findings reveal a new mechanism that promotes the emergence of spontaneous OX R mutants in bacterial organisms. IMPORTANCE Although a mutator phenotype has been shown to promote antibiotic resistance in many bacterial species, little is known about its contribution to the emergence of OX R mutants. This work describes the link between a mutator phenotype and the enhanced emergence of OX R mutants as well as its underlying mechanism involving DNA repair and mutations in PerR. Since DNA repair systems and PerR are well conserved in many bacterial species, especially in Gram positives, the same mechanism may operate in multiple bacterial species. Additionally, we developed a novel method that allows for rapid quantification of spontaneous OX R mutants in a bacterial population. This method represents a technical

  10. Cooling of hot bubbles by surface texture during the boiling crisis

    NASA Astrophysics Data System (ADS)

    Dhillon, Navdeep; Buongiorno, Jacopo; Varanasi, Kripa

    2015-11-01

    We report the existence of maxima in critical heat flux (CHF) enhancement for pool boiling on textured hydrophilic surfaces and reveal the interaction mechanism between bubbles and surface texture that governs the boiling crisis phenomenon. Boiling is a process of fundamental importance in many engineering and industrial applications but the maximum heat flux that can be absorbed by the boiling liquid (or CHF) is limited by the boiling crisis. Enhancing the CHF of industrial boilers by surface texturing can lead to substantial energy savings and reduction in greenhouse gas emissions on a global scale. However, the fundamental mechanisms behind this enhancement are not well understood, with some previous studies indicating that CHF should increase monotonically with increasing texture density. However, using pool boiling experiments on a parametrically designed set of plain and nano-textured micropillar surfaces, we show that there is an optimum intermediate texture density that maximizes CHF and further that the length scale of this texture is of fundamental significance. Using imbibition experiments and high-speed optical and infrared imaging, we reveal the fundamental mechanisms governing the CHF enhancement maxima in boiling crisis. We acknowledge funding from the Chevron corporation.

  11. Microbiological effectiveness of disinfecting water by boiling in rural Guatemala.

    PubMed

    Rosa, Ghislaine; Miller, Laura; Clasen, Thomas

    2010-03-01

    Boiling is the most common means of treating water in the home and the benchmark against which alternative point-of-use water treatment options must be compared. In a 5-week study in rural Guatemala among 45 households who claimed they always or almost always boiled their drinking water, boiling was associated with a 86.2% reduction in geometric mean thermotolerant coliforms (TTC) (N = 206, P < 0.0001). Despite consistent levels of fecal contamination in source water, 71.2% of stored water samples from self-reported boilers met the World Health Organization guidelines for safe drinking water (0 TTC/100 mL), and 10.7% fell within the commonly accepted low-risk category of (1-10 TTC/100 mL). As actually practiced in the study community, boiling significantly improved the microbiological quality of drinking water, though boiled and stored drinking water is not always free of fecal contaminations.

  12. Hydrogenase activity in the foodborne pathogen Campylobacter jejuni depends upon a novel ABC-type nickel transporter (NikZYXWV) and is SlyD-independent.

    PubMed

    Howlett, Robert M; Hughes, Bethan M; Hitchcock, Andrew; Kelly, David J

    2012-06-01

    Campylobacter jejuni is a human pathogen of worldwide significance. It is commensal in the gut of many birds and mammals, where hydrogen is a readily available electron donor. The bacterium possesses a single membrane-bound, periplasmic-facing NiFe uptake hydrogenase that depends on the acquisition of environmental nickel for activity. The periplasmic binding protein Cj1584 (NikZ) of the ATP binding cassette (ABC) transporter encoded by the cj1584c-cj1580c (nikZYXWV) operon in C. jejuni strain NCTC 11168 was found to be nickel-repressed and to bind free nickel ions with a submicromolar K(d) value, as measured by fluorescence spectroscopy. Unlike the Escherichia coli NikA protein, NikZ did not bind EDTA-chelated nickel and lacks key conserved residues implicated in metallophore interaction. A C. jejuni cj1584c null mutant strain showed an approximately 22-fold decrease in intracellular nickel content compared with the wild-type strain and a decreased rate of uptake of (63)NiCl(2). The inhibition of residual nickel uptake at higher nickel concentrations in this mutant by hexa-ammine cobalt (III) chloride or magnesium ions suggests that low-affinity uptake occurs partly through the CorA magnesium transporter. Hydrogenase activity was completely abolished in the cj1584c mutant after growth in unsupplemented media, but was fully restored after growth with 0.5 mM nickel chloride. Mutation of the putative metallochaperone gene slyD (cj0115) had no effect on either intracellular nickel accumulation or hydrogenase activity. Our data reveal a strict dependence of hydrogenase activity in C. jejuni on high-affinity nickel uptake through an ABC transporter that has distinct properties compared with the E. coli Nik system.

  13. The boiling point of stratospheric aerosols.

    NASA Technical Reports Server (NTRS)

    Rosen, J. M.

    1971-01-01

    A photoelectric particle counter was used for the measurement of aerosol boiling points. The operational principle involves raising the temperature of the aerosol by vigorously heating a portion of the intake tube. At or above the boiling point, the particles disintegrate rather quickly, and a noticeable effect on the size distribution and concentration is observed. Stratospheric aerosols appear to have the same volatility as a solution of 75% sulfuric acid. Chemical analysis of the aerosols indicates that there are other substances present, but that the sulfate radical is apparently the major constituent.

  14. Campylobacter protein oxidation influences epithelial cell invasion or intracellular survival as well as intestinal tract colonization in chickens.

    PubMed

    Lasica, A M; Wyszynska, A; Szymanek, K; Majewski, P; Jagusztyn-Krynicka, E K

    2010-01-01

    The Dsb family of redox proteins catalyzes disulfide bond formation and isomerization. Since mutations in dsb genes change the conformation and stability of many extracytoplasmic proteins, and since many virulence factors of pathogenic bacteria are extracytoplasmic, inactivation of dsb genes often results in pathogen attenuation. This study investigated the role of 2 membrane-bound oxidoreductases, DsbB and DsbI, in the Campylobacter jejuni oxidative Dsb pathway. Campylobacter mutants, lacking DsbB or DsbI or both, were constructed by allelic replacement and used in the human intestinal epithelial T84 cell line for the gentamicin protection assay (invasion assay) and chicken colonization experiments. In C. coli strain 23/1, the inactivation of the dsbB or dsbI gene separately did not significantly affect the colonization process. However, simultaneous disruption of both membrane-bound oxidoreductase genes significantly decreased the strain’s ability to colonize chicken intestines. Moreover, C. jejuni strain 81-176 with mutated dsbB or dsbI genes showed reduced invasion/intracellular survival abilities. No cells of the double mutants (dsbB⁻ dsbI⁻) of C. jejuni 81-176 were recovered from human cells after 3 h of invasion.

  15. Microbiological Effectiveness of Disinfecting Water by Boiling in Rural Guatemala

    PubMed Central

    Rosa, Ghislaine; Miller, Laura; Clasen, Thomas

    2010-01-01

    Boiling is the most common means of treating water in the home and the benchmark against which alternative point-of-use water treatment options must be compared. In a 5-week study in rural Guatemala among 45 households who claimed they always or almost always boiled their drinking water, boiling was associated with a 86.2% reduction in geometric mean thermotolerant coliforms (TTC) (N = 206, P < 0.0001). Despite consistent levels of fecal contamination in source water, 71.2% of stored water samples from self-reported boilers met the World Health Organization guidelines for safe drinking water (0 TTC/100 mL), and 10.7% fell within the commonly accepted low-risk category of (1–10 TTC/100 mL). As actually practiced in the study community, boiling significantly improved the microbiological quality of drinking water, though boiled and stored drinking water is not always free of fecal contaminations. PMID:20207876

  16. BOILING REACTORS

    DOEpatents

    Untermyer, S.

    1962-04-10

    A boiling reactor having a reactivity which is reduced by an increase in the volume of vaporized coolant therein is described. In this system unvaporized liquid coolant is extracted from the reactor, heat is extracted therefrom, and it is returned to the reactor as sub-cooled liquid coolant. This reduces a portion of the coolant which includes vaporized coolant within the core assembly thereby enhancing the power output of the assembly and rendering the reactor substantially self-regulating. (AEC)

  17. A Cytolethal Distending Toxin Gene-Based Multiplex PCR Assay for Campylobacter jejuni, C. fetus, C. coli, C. upsaliensis, C. hyointestinalis, and C. lari.

    PubMed

    Kamei, Kazumasa; Kawabata, Hiroki; Asakura, Masahiro; Samosornsuk, Worada; Hinenoya, Atsushi; Nakagawa, Shinsaku; Yamasaki, Shinji

    2016-05-20

    In this study, we devised a multiplex PCR assay based on the gene of cytolethal distending toxin (cdt) B subunit to simultaneously detect and discriminate Campylobacter jejuni, C. fetus, C. coli, C. upsaliensis, C. hyointestinalis, and C. lari. Species-specific PCR products were successfully obtained from all 38 C. jejuni, 12 C. fetus, 39 C. coli, 22 C. upsaliensis, 24 C. hyointestinalis, and 7 C. lari strains tested. On the other hand, no specific PCR products were obtained from other campylobacters and bacterial species tested (41 strains in total). The proposed multiplex PCR assay is a valuable tool for detection and descrimination of 6 major Campylobacter species, that are associated with gastrointestinal diseases in humans.

  18. Experimental evidence of the vapor recoil mechanism in the boiling crisis.

    PubMed

    Nikolayev, V S; Chatain, D; Garrabos, Y; Beysens, D

    2006-11-03

    Boiling crisis experiments are carried out in the vicinity of the liquid-gas critical point of H2. A magnetic gravity compensation setup is used to enable nucleate boiling at near critical pressure. The measurements of the critical heat flux that defines the threshold for the boiling crisis are carried out as a function of the distance from the critical point. The obtained power law behavior and the boiling crisis dynamics agree with the predictions of the vapor recoil mechanism and disagree with the classical vapor column mechanism.

  19. Full evaporation headspace gas chromatography for sensitive determination of high boiling point volatile organic compounds in low boiling matrices.

    PubMed

    Mana Kialengila, Didi; Wolfs, Kris; Bugalama, John; Van Schepdael, Ann; Adams, Erwin

    2013-11-08

    Determination of volatile organic components (VOC's) is often done by static headspace gas chromatography as this technique is very robust and combines easy sample preparation with good selectivity and low detection limits. This technique is used nowadays in different applications which have in common that they have a dirty matrix which would be problematic in direct injection approaches. Headspace by nature favors the most volatile compounds, avoiding the less volatile to reach the injector and column. As a consequence, determination of a high boiling solvent in a lower boiling matrix becomes challenging. Determination of VOCs like: xylenes, cumene, N,N-dimethylformamide (DMF), dimethyl sulfoxide (DMSO), N,N-dimethylacetamide (DMA), N-methyl-2-pyrrolidone (NMP), 1,3-dimethyl-2-imidazolidinone (DMI), benzyl alcohol (BA) and anisole in water or water soluble products are an interesting example of the arising problems. In this work, a headspace variant called full evaporation technique is worked out and validated for the mentioned solvents. Detection limits below 0.1 μg/vial are reached with RSD values below 10%. Mean recovery values ranged from 92.5 to 110%. The optimized method was applied to determine residual DMSO in a water based cell culture and DMSO and DMA in tetracycline hydrochloride (a water soluble sample). Copyright © 2013 Elsevier B.V. All rights reserved.

  20. Heat Transfer in Boiling Dilute Emulsion with Strong Buoyancy

    NASA Astrophysics Data System (ADS)

    Freeburg, Eric Thomas

    Little attention has been given to the boiling of emulsions compared to that of boiling in pure liquids. The advantages of using emulsions as a heat transfer agent were first discovered in the 1970s and several interesting features have since been studied by few researchers. Early research focuses primarily on pool and flow boiling and looks to determine a mechanism by which the boiling process occurs. This thesis looks at the boiling of dilute emulsions in fluids with strong buoyant forces. The boiling of dilute emulsions presents many favorable characteristics that make it an ideal agent for heat transfer. High heat flux electronics, such as those seen in avionics equipment, produce high heat fluxes of 100 W/cm2 or more, but must be maintained at low temperatures. So far, research on single phase convection and flow boiling in small diameter channels have yet to provide an adequate solution. Emulsions allow the engineer to tailor the solution to the specific problem. The fluid can be customized to retain the high thermal conductivity and specific heat capacity of the continuous phase while enhancing the heat transfer coefficient through boiling of the dispersed phase component. Heat transfer experiments were carried out with FC-72 in water emulsions. FC-72 has a saturation temperature of 56 °C, far below that of water. The parameters were varied as follows: 0% ≤ epsilon ≤ 1% and 1.82 x 1012 ≤ RaH ≤ 4.42 x 1012. Surface temperatures along the heated surface reached temperature that were 20 °C in excess of the dispersed phase saturation temperature. An increase of ˜20% was seen in the average Nusselt numbers at the highest Rayleigh numbers. Holography was used to obtain images of individual and multiple FC-72 droplets in the boundary layer next to the heated surface. The droplet diameters ranged from 0.5 mm to 1.3 mm. The Magnus effect was observed when larger individual droplets were injected into the boundary layer, causing the droplets to be pushed

  1. Numerical and Experimental Study of Mechanisms Involved in Boiling Histotripsy.

    PubMed

    Pahk, Ki Joo; Gélat, Pierre; Sinden, David; Dhar, Dipok Kumar; Saffari, Nader

    2017-12-01

    The aim of boiling histotripsy is to mechanically fractionate tissue as an alternative to thermal ablation for therapeutic applications. In general, the shape of a lesion produced by boiling histotripsy is tadpole like, consisting of a head and a tail. Although many studies have demonstrated the efficacy of boiling histotripsy for fractionating solid tumors, the exact mechanisms underpinning this phenomenon are not yet well understood, particularly the interaction of a boiling vapor bubble with incoming incident shockwaves. To investigate the mechanisms involved in boiling histotripsy, a high-speed camera with a passive cavitation detection system was used to observe the dynamics of bubbles produced in optically transparent tissue-mimicking gel phantoms exposed to the field of a 2.0-MHz high-intensity focused ultrasound (HIFU) transducer. We observed that boiling bubbles were generated in a localized heated region and cavitation clouds were subsequently induced ahead of the expanding bubble. This process was repeated with HIFU pulses and eventually resulted in a tadpole-shaped lesion. A simplified numerical model describing the scattering of the incident ultrasound wave by a vapor bubble was developed to help interpret the experimental observations. Together with the numerical results, these observations suggest that the overall size of a lesion induced by boiling histotripsy is dependent on the sizes of (i) the heated region at the HIFU focus and (ii) the backscattered acoustic field by the original vapor bubble. Copyright © 2017 World Federation for Ultrasound in Medicine and Biology. Published by Elsevier Inc. All rights reserved.

  2. Acoustic emission feedback control for control of boiling in a microwave oven

    DOEpatents

    White, Terry L.

    1991-01-01

    An acoustic emission based feedback system for controlling the boiling level of a liquid medium in a microwave oven is provided. The acoustic emissions from the medium correlated with surface boiling is used to generate a feedback control signal proportional to the level of boiling of the medium. This signal is applied to a power controller to automatically and continuoulsly vary the power applied to the oven to control the boiling at a selected level.

  3. Single-bubble boiling under Earth's and low gravity

    NASA Astrophysics Data System (ADS)

    Khusid, Boris; Elele, Ezinwa; Lei, Qian; Tang, John; Shen, Yueyang

    2017-11-01

    Miniaturization of electronic systems in terrestrial and space applications is challenged by a dramatic increase in the power dissipation per unit volume with the occurrence of localized hot spots where the heat flux is much higher than the average. Cooling by forced gas or liquid flow appears insufficient to remove high local heat fluxes. Boiling that involves evaporation of liquid in a hot spot and condensation of vapor in a cold region can remove a significantly larger amount of heat through the latent heat of vaporization than force-flow cooling can carry out. Traditional methods for enhancing boiling heat transfer in terrestrial and space applications focus on removal of bubbles from the heating surface. In contrast, we unexpectedly observed a new boiling regime of water under Earth's gravity and low gravity in which a bubble was pinned on a small heater up to 270°C and delivered a heat flux up to 1.2 MW/m2 that was as high as the critical heat flux in the classical boiling regime on Earth .Low gravity measurements conducted in parabolic flights in NASA Boeing 727. The heat flux in flight and Earth's experiments was found to rise linearly with increasing the heater temperature. We will discuss physical mechanisms underlying heat transfer in single-bubble boiling. The work supported by NASA Grants NNX12AM26G and NNX09AK06G.

  4. Boiling-induced formation of colloidal gold in black smoker hydrothermal fluids

    USGS Publications Warehouse

    Gartman, Amy; Hannington, Mark; Jamieson, John W.; Peterkin, Ben; Garbe-Schönberg, Dieter; Findlay, Alyssa J; Fuchs, Sebastian; Kwasnitschka, Tom

    2017-01-01

    Gold colloids occur in black smoker fluids from the Niua South hydrothermal vent field, Lau Basin (South Pacific Ocean), confirming the long-standing hypothesis that gold may undergo colloidal transport in hydrothermal fluids. Six black smoker vents, varying in temperature from 250 °C to 325 °C, were sampled; the 325 °C vent was boiling at the time of sampling and the 250 °C fluids were diffusely venting. Native gold particles ranging from <50 nm to 2 µm were identified in 4 of the fluid samples and were also observed to precipitate on the sampler during collection from the boiling vent. Total gold concentrations (dissolved and particulate) in the fluid samples range from 1.6 to 5.4 nM in the high-temperature, focused flow vents. Although the gold concentrations in the focused flow fluids are relatively high, they are lower than potential solubilities prior to boiling and indicate that precipitation was boiling induced, with sulfide lost upon boiling to exsolution and metal sulfide formation. Gold concentrations reach 26.7 nM in the 250 °C diffuse flow sample, and abundant native gold particles were also found in the fluids and associated sulfide chimney and are interpreted to be a product of colloid accumulation and growth following initial precipitation upon boiling. These results indicate that colloid-driven precipitation as a result of boiling, the persistence of colloids after boiling, and the accumulation of colloids in diffuse flow fluids are important mechanisms for the enrichment of gold in seafloor hydrothermal systems.

  5. Multistate outbreak of Campylobacter jejuni infections associated with undercooked chicken livers--northeastern United States, 2012.

    PubMed

    2013-11-08

    In October 2012 the Vermont Department of Health (VDH) identified three cases of laboratory-confirmed Campylobacter jejuni infection in Vermont residents; the isolates had indistinguishable pulsed-field gel electrophoresis (PFGE) patterns. A query of PulseNet, the national molecular subtyping network for foodborne disease surveillance, led to the identification of one additional case each from New Hampshire, New York, and Vermont that had been reported in the preceding 6 months. An investigation led by VDH found that all six patients had been exposed to raw or lightly cooked chicken livers that had been produced at the same Vermont poultry establishment (establishment A). Livers collected from this establishment yielded the outbreak strain of C. jejuni. In response, establishment A voluntarily ceased the sale of chicken livers on November 9. A food safety assessment conducted by the U.S. Department of Agriculture's Food Safety and Inspection Service (USDA-FSIS) found no major violations at the establishment. This is the first reported multistate outbreak of campylobacteriosis associated with chicken liver in the United States. Public health professionals, members of the food industry, and consumers should be aware that chicken livers often are contaminated with Campylobacter and that fully cooking products made with chicken liver is the only way to prepare them so they are safe to eat.

  6. Steady State Film Boiling Heat Transfer Simulated With Trace V4.160

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Audrius Jasiulevicius; Rafael Macian-Juan

    2006-07-01

    This paper presents the results of the assessment and analysis of TRACE v4.160 heat transfer predictions in the post-CHF (critical heat flux) region and discusses the possibilities to improve the TRACE v4.160 code predictions in the film boiling heat transfer when applying different film boiling correlations. For this purpose, the TRACE v4.160-calculated film boiling heat flux and the resulting maximum inner wall temperatures during film boiling in single tubes were compared with experimental data obtained at the Royal Institute of Technology (KTH) in Stockholm, Sweden. The experimental database included measurements for pressures ranging from 30 to 200 bar and coolantmore » mass fluxes from 500 to 3000 kg/m{sup 2}s. It was found that TRACE v4.160 does not produce correct predictions of the film boiling heat flux, and consequently of the maximum inner wall temperature in the test section, under the wide range of conditions documented in the KTH experiments. In particular, it was found that the standard TRACE v4.160 under-predicts the film boiling heat transfer coefficient at low pressure-low mass flux and high pressure-high mass flux conditions. For most of the rest of the investigated range of parameters, TRACE v4.160 over-predicts the film boiling heat transfer coefficient, which can lead to non-conservative predictions in applications to nuclear power plant analyses. Since no satisfactory agreement with the experimental database was obtained with the standard TRACE v4.160 film boiling heat transfer correlations, we have added seven film boiling correlations to TRACE v4.160 in order to investigate the possibility to improve the code predictions for the conditions similar to the KTH tests. The film boiling correlations were selected among the most commonly used film boiling correlations found in the open literature, namely Groeneveld 5.7, Bishop (2 correlations), Tong, Konkov, Miropolskii and Groeneveld-Delorme correlations. The only correlation among the

  7. 20. RW Meyer Sugar Mill: 18761889. Boiling House Interior, 1878. ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    20. RW Meyer Sugar Mill: 1876-1889. Boiling House Interior, 1878. View: Remains of south wall. The molasses storage pits are below the floor in the foreground. The remaining piece of floor indicates the form of the entire floor. The sorghum pan and boiling range flue slope from left to right (east to west) and permitted batches of cane juice to flow through the boiling pan by gravity. The beams, joists, truss work are built of northwest pine. The sides and floor boards are built of redwood. The boiling range flue is built of fire-brick, masonry, and portland cement. The corrugated roof appears to be a later addition, not contemporary with mill operation. - R. W. Meyer Sugar Mill, State Route 47, Kualapuu, Maui County, HI

  8. Effects of boiling and in vitro gastrointestinal digestion on the antioxidant activity of Sonchus oleraceus leaves.

    PubMed

    Mawalagedera, S M M R; Ou, Zong-Quan; McDowell, Arlene; Gould, Kevin S

    2016-03-01

    Leaves of Sonchus oleraceus L. are especially rich in phenolic compounds and have potent extractable antioxidants. However, it is not known how their antioxidant activity changes after cooking and gastrointestinal digestion. We recorded the profile of phenolics and their associated antioxidant activity in both raw and boiled S. oleraceus leaf extracts after in vitro gastric and intestinal digestion, and quantified their antioxidant potentials using Caco-2 and HepG2 cells. Boiling significantly diminished the oxygen radical absorbance capacity (ORAC) and concentrations of ascorbate and chicoric acid in the soluble fractions. In contrast, 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging and concentrations of caftaric and chlorogenic acids were unaffected. Phenolics in the soluble fraction were absorbed into cultured human cells and exerted antioxidant activity. Only chlorogenic acid content remained stable during gastrointestinal digestion. S. oleraceus appears to be an excellent dietary source of phenolic antioxidants.

  9. Campylobacter jejuni dsb gene expression is regulated by iron in a Fur-dependent manner and by a translational coupling mechanism.

    PubMed

    Grabowska, Anna D; Wandel, Michał P; Łasica, Anna M; Nesteruk, Monika; Roszczenko, Paula; Wyszyńska, Agnieszka; Godlewska, Renata; Jagusztyn-Krynicka, Elzbieta K

    2011-07-25

    Many bacterial extracytoplasmic proteins are stabilized by intramolecular disulfide bridges that are formed post-translationally between their cysteine residues. This protein modification plays an important role in bacterial pathogenesis, and is facilitated by the Dsb (disulfide bond) family of the redox proteins. These proteins function in two parallel pathways in the periplasmic space: an oxidation pathway and an isomerization pathway. The Dsb oxidative pathway in Campylobacter jejuni is more complex than the one in the laboratory E. coli K-12 strain. In the C. jejuni 81-176 genome, the dsb genes of the oxidative pathway are arranged in three transcriptional units: dsbA2-dsbB-astA, dsbA1 and dba-dsbI. Their transcription responds to an environmental stimulus - iron availability - and is regulated in a Fur-dependent manner. Fur involvement in dsb gene regulation was proven by a reporter gene study in a C. jejuni wild type strain and its isogenic fur mutant. An electrophoretic mobility shift assay (EMSA) confirmed that analyzed genes are members of the Fur regulon but each of them is regulated by a disparate mechanism, and both the iron-free and the iron-complexed Fur are able to bind in vitro to the C. jejuni promoter regions. This study led to identification of a new iron- and Fur-regulated promoter that drives dsbA1 gene expression in an indirect way. Moreover, the present work documents that synthesis of DsbI oxidoreductase is controlled by the mechanism of translational coupling. The importance of a secondary dba-dsbI mRNA structure for dsbI mRNA translation was verified by estimating individual dsbI gene expression from its own promoter. The present work shows that iron concentration is a significant factor in dsb gene transcription. These results support the concept that iron concentration - also through its influence on dsb gene expression - might control the abundance of extracytoplasmic proteins during different stages of infection. Our work further shows

  10. Early Onset of Nucleate Boiling on Gas-covered Biphilic Surfaces.

    PubMed

    Shen, Biao; Yamada, Masayuki; Hidaka, Sumitomo; Liu, Jiewei; Shiomi, Junichiro; Amberg, Gustav; Do-Quang, Minh; Kohno, Masamichi; Takahashi, Koji; Takata, Yasuyuki

    2017-05-17

    For phase-change cooling schemes for electronics, quick activation of nucleate boiling helps safeguard the electronics components from thermal shocks associated with undesired surface superheating at boiling incipience, which is of great importance to the long-term system stability and reliability. Previous experimental studies show that bubble nucleation can occur surprisingly early on mixed-wettability surfaces. In this paper, we report unambiguous evidence that such unusual bubble generation at extremely low temperatures-even below the boiling point-is induced by a significant presence of incondensable gas retained by the hydrophobic surface, which exhibits exceptional stability even surviving extensive boiling deaeration. By means of high-speed imaging, it is revealed that the consequently gassy boiling leads to unique bubble behaviour that stands in sharp contrast with that of pure vapour bubbles. Such findings agree qualitatively well with numerical simulations based on a diffuse-interface method. Moreover, the simulations further demonstrate strong thermocapillary flows accompanying growing bubbles with considerable gas contents, which is associated with heat transfer enhancement on the biphilic surface in the low-superheat region.

  11. Structural analysis of PseH, the Campylobacter jejuni N-acetyltransferase involved in bacterial O-linked glycosylation

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Song, Wan Seok; Nam, Mi Sun; Namgung, Byeol

    2015-03-20

    Campylobacter jejuni is a bacterium that uses flagella for motility and causes worldwide acute gastroenteritis in humans. The C. jejuni N-acetyltransferase PseH (cjPseH) is responsible for the third step in flagellin O-linked glycosylation and plays a key role in flagellar formation and motility. cjPseH transfers an acetyl group from an acetyl donor, acetyl coenzyme A (AcCoA), to the amino group of UDP-4-amino-4,6-dideoxy-N-acetyl-β-L-altrosamine to produce UDP-2,4-diacetamido-2,4,6-trideoxy-β-L-altropyranose. To elucidate the catalytic mechanism of cjPseH, crystal structures of cjPseH alone and in complex with AcCoA were determined at 1.95 Å resolution. cjPseH folds into a single-domain structure of a central β-sheet decorated by four α-helicesmore » with two continuously connected grooves. A deep groove (groove-A) accommodates the AcCoA molecule. Interestingly, the acetyl end of AcCoA points toward an open space in a neighboring shallow groove (groove-S), which is occupied by extra electron density that potentially serves as a pseudosubstrate, suggesting that the groove-S may provide a substrate-binding site. Structure-based comparative analysis suggests that cjPseH utilizes a unique catalytic mechanism of acetylation that has not been observed in other glycosylation-associated acetyltransferases. Thus, our studies on cjPseH will provide valuable information for the design of new antibiotics to treat C. jejuni-induced gastroenteritis. - Highlights: • cjPseH adopts a single-domain structure of a central β-sheet decorated by α-helices. • cjPseH features two continuously connected grooves on the protein surface. • Acetyl coenzyme A (AcCoA) binds into a deep groove of cjPseH in an ‘L’ shape. • The acetyl end of AcCoA points to a wide groove, a potential substrate-binding site.« less

  12. Major contribution of the type II beta carbonic anhydrase CanB (Cj0237) to the capnophilic growth phenotype of Campylobacter jejuni.

    PubMed

    Al-Haideri, Halah; White, Michael A; Kelly, David J

    2016-02-01

    Campylobacter jejuni, the leading cause of human bacterial gastroenteritis, requires low environmental oxygen and high carbon dioxide for optimum growth, but the molecular basis for the carbon dioxide requirement is unclear. One factor may be inefficient conversion of gaseous CO2 to bicarbonate, the required substrate of various carboxylases. Two putative carbonic anhydrases (CAs) are encoded in the genome of C. jejuni strain NCTC 11168 (Cj0229 and Cj0237). Here, we show that the deletion of the cj0237 (canB) gene alone prevents growth in complex media at low (1% v/v) CO2 and significantly reduces the growth rate at high (5% v/v) CO2. In minimal media incubated under high CO2, the canB mutant grew on L-aspartate but not on the key C3 compounds L-serine, pyruvate and L-lactate, showing that CanB is crucial in bicarbonate provision for pyruvate carboxylase-mediated oxaloacetate synthesis. Nevertheless, purified CanB (a dimeric, anion and acetazolamide sensitive, zinc-containing type II beta-class enzyme) hydrates CO2 actively only above pH 8 and with a high Km (∼ 34 mM). At typical cytoplasmic pH values and low CO2, these kinetic properties might limit intracellular bicarbonate availability. Taken together, our data suggest CanB is a major contributor to the capnophilic growth phenotype of C. jejuni. © 2015 Society for Applied Microbiology and John Wiley & Sons Ltd.

  13. Boiling local heat transfer enhancement in minichannels using nanofluids

    PubMed Central

    2013-01-01

    This paper reports an experimental study on nanofluid convective boiling heat transfer in parallel rectangular minichannels of 800 μm hydraulic diameter. Experiments are conducted with pure water and silver nanoparticles suspended in water base fluid. Two small volume fractions of silver nanoparticles suspended in water are tested: 0.000237% and 0.000475%. The experimental results show that the local heat transfer coefficient, local heat flux, and local wall temperature are affected by silver nanoparticle concentration in water base fluid. In addition, different correlations established for boiling flow heat transfer in minichannels or macrochannels are evaluated. It is found that the correlation of Kandlikar and Balasubramanian is the closest to the water boiling heat transfer results. The boiling local heat transfer enhancement by adding silver nanoparticles in base fluid is not uniform along the channel flow. Better performances and highest effect of nanoparticle concentration on the heat transfer are obtained at the minichannels entrance. PMID:23506445

  14. 46 CFR 154.705 - Cargo boil-off as fuel: General.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... STANDARDS FOR SELF-PROPELLED VESSELS CARRYING BULK LIQUEFIED GASES Design, Construction and Equipment Cargo Pressure and Temperature Control § 154.705 Cargo boil-off as fuel: General. (a) Each cargo boil-off fuel...

  15. 46 CFR 154.705 - Cargo boil-off as fuel: General.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... STANDARDS FOR SELF-PROPELLED VESSELS CARRYING BULK LIQUEFIED GASES Design, Construction and Equipment Cargo Pressure and Temperature Control § 154.705 Cargo boil-off as fuel: General. (a) Each cargo boil-off fuel...

  16. 46 CFR 154.705 - Cargo boil-off as fuel: General.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... STANDARDS FOR SELF-PROPELLED VESSELS CARRYING BULK LIQUEFIED GASES Design, Construction and Equipment Cargo Pressure and Temperature Control § 154.705 Cargo boil-off as fuel: General. (a) Each cargo boil-off fuel...

  17. 46 CFR 154.705 - Cargo boil-off as fuel: General.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... STANDARDS FOR SELF-PROPELLED VESSELS CARRYING BULK LIQUEFIED GASES Design, Construction and Equipment Cargo Pressure and Temperature Control § 154.705 Cargo boil-off as fuel: General. (a) Each cargo boil-off fuel...

  18. 46 CFR 154.705 - Cargo boil-off as fuel: General.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... STANDARDS FOR SELF-PROPELLED VESSELS CARRYING BULK LIQUEFIED GASES Design, Construction and Equipment Cargo Pressure and Temperature Control § 154.705 Cargo boil-off as fuel: General. (a) Each cargo boil-off fuel...

  19. Mechanisms underlying zoonotic success of Campylobacter jejuni: the CprRS two-component regulatory system influences essential processes, biofilm formation, and pathogenesis

    USDA-ARS?s Scientific Manuscript database

    Campylobacter jejuni is a leading cause of food- and waterbourne bacterial gastroenteritis in the developed world. Although illness is usually self-limiting, immunocompromised individuals are at risk for infections recalcitrant to antibiotic treatment, and prior campylobacter infection correlates wi...

  20. Film Boiling Heat Transfer Properties of Liquid Hydrogen in Natural Convection

    NASA Astrophysics Data System (ADS)

    Horie, Y.; Shirai, Y.; Shiotsu, M.; Matsuzawa, T.; Yoneda, K.; Shigeta, H.; Tatsumoto, H.; Hata, K.; Naruo, Y.; Kobayashi, H.; Inatani, Y.

    Film boiling heat transfer properties of LH2 for various pressures and subcooling conditions were measured by applying electric current to give an exponential heat input to a PtCo wire with a diameter of 1.2 mm submerged in LH2. The heated wire was set to be horizontal to the ground. The heat transfer coefficient in the film boiling region was higher for higher pressure and higher subcooling. The experimental results are compared with the equation of pool film boiling heat transfer. It is confirmed that the pool film boiling heat transfer coefficients in LH2 can be expressed by this equation.

  1. Marangoni Effects in the Boiling of Binary Fluid Mixtures

    NASA Technical Reports Server (NTRS)

    Ahmed, Sayeed; Carey, Van P.; Motil, Brian

    1996-01-01

    Results of very recent experimental studies indicate that during nucleate boiling in some binary mixture, Marangoni effects augment the gravity driven flow of liquid towards the heated surface. With gravity present, it is impossible to separate the two effects. The reduced gravity environment gives an unique opportunity to explore th role of Marangoni effects on the boiling mechanisms free of gravitational body forces that obscure the role of such effects. However, recent experimental results suggest that under reduced gravity conditions, Marangoni effects is the dominant mechanism of vapor-liquid exchange at the surface for some binary mixture. To further explore such effects, experiments have been conducted with water/2-propanol mixtures at three different concentrations under normal gravity with different orientations of the heater surface and under reduce gravity aboard the DC-9 aircraft at NASA Lewis Research Center. The system pressure was sub atmospheric (approx. 8 kP at 1g(n)) and the bulk liquid temperature varied from low subcooling to near saturation. The molar concentrations of 2-propanol tested were 0.015, 0.025, and 0.1. Boiling curves were obtained both for high gravity (approx. 2g(n)) and reduce gravity (approx. 0.01g(n)). For each concentration of 2-propanol, the critical heat flux has been determined in the flight experiments only for reduced gravity conditions. Comparison of boiling curves and CHF obtained under l-g(n) an reduced gravity indicates that boiling mechanism in this mixtures is nearly independent of gravity. The results also indicate that the Marangoni mechanism is strong enough in these mixtures to sustain the boiling under reduced gravity conditions.

  2. Reduced Boil-Off System Sizing

    NASA Technical Reports Server (NTRS)

    Guzik, Monica C.; Plachta, David W.; Feller, Jeffrey R.

    2015-01-01

    NASA is currently developing cryogenic propellant storage and transfer systems for future space exploration and scientific discovery missions by addressing the need to raise the technology readiness level of cryogenic fluid management technologies. Cryogenic propellants are baselined in many propulsion systems due to their inherently high specific impulse; however, their low boiling points can cause substantial boil-off losses over time. Recent efforts such as the Reduced Boil-off Testing and the Active Thermal Control Scaling Study provide important information on the benefit of an active cooling system applied to LH2 propellant storage. Findings show that zero-boil off technologies can reduce overall mass in LH2 storage systems when low Earth orbit loiter periods extend beyond two months. A significant part of this mass reduction is realized by integrating two stages of cooling: a 20 K stage to intercept heat at the tank surface, and a 90 K stage to reduce the heat entering the less efficient 20 K stage. A missing element in previous studies, which is addressed in this paper, is the development of a direct method for sizing the 90 K cooling stage. Such a method requires calculation of the heat entering both the 90 K and 20 K stages as compared to the overall system masses, and is reliant upon the temperature distribution, performance, and unique design characteristics of the system in question. By utilizing the known conductance of a system without active thermal control, the heat being intercepted by a 90 K stage can be calculated to find the resultant lift and mass of each active thermal control stage. Integral to this is the thermal conductance of the cooling straps and the broad area cooling shield, key parts of the 90 K stage. Additionally, a trade study is performed to show the ability of the 90 K cooling stage to reduce the lift on the 20 K cryocooler stage, which is considerably less developed and efficient than 90 K cryocoolers.

  3. Nucleate pool boiling in subcooled liquid under microgravity: Results of TEXUS experimental investigations

    NASA Astrophysics Data System (ADS)

    Zell, M.; Straub, J.; Weinzierl, A.

    1984-12-01

    Experiments on subcooled nucleate pool boiling in microgravity were carried out to separate gravity driven effects on heat transfer within the boiling process. A ballistic trajectory by sounding rocket flight (TEXUS 5 and 10) achieved a gravity level of a/g = 0.0001 for 360 sec. For determination of geometrical effects on heat transport two different experimental configurations (platinum wire and flat plate) were employed. Boiling curves and bubble dynamics recorded by cinematography lead to gravity independent modelling of the boiling phenomena. The results ensure the applicability and high efficiency of nucleate pool boiling for heat exchangers in space laboratories.

  4. Zero Boil-OFF Tank Hardware Setup

    NASA Image and Video Library

    2017-09-19

    iss053e027051 (Sept. 19, 2017) --- Flight Engineer Joe Acaba works in the U.S. Destiny laboratory module setting up hardware for the Zero Boil-Off Tank (ZBOT) experiment. ZBOT uses an experimental fluid to test active heat removal and forced jet mixing as alternative means for controlling tank pressure for volatile fluids. Rocket fuel, spacecraft heating and cooling systems, and sensitive scientific instruments rely on very cold cryogenic fluids. Heat from the environment around cryogenic tanks can cause their pressures to rise, which requires dumping or "boiling off" fluid to release the excess pressure, or actively cooling the tanks in some way.

  5. The Boiling eXperiment Facility (BXF) for the Microgravity Science Glovebox (MSG)

    NASA Technical Reports Server (NTRS)

    McQuillen, John; Chao, David; Vergilii, Frank

    2006-01-01

    Boiling is an effective means of cooling by removing heat from surfaces through vaporization of a working fluid. It is also affected by both the magnitude and direction of gravity. By conducting pool boiling tests in microgravity, the effect of buoyancy n the overall boiling process and the relative magnitude of other phenomena can be assessed. The Boiling eXperiment Facility (BXF) is being built for the Microgravity Science Glovebox. This facility will conduct two pool boiling studies. The first study the Microheater Array Boiling Experiment (MABE) uses two 96 element microheater arrays, 2.7 mm and 7.0 mm in size, to measure localized hear fluxes while operating at a constant temperature. The other experiment, the Nucleate Pool Boiling eXperiment (NPBX) uses a 85 mm diameter heater wafer that has been "seeded" with five individually-controlled nucleation sites to study bubble nucleation, growth, coalescence and departure. The BXF uses normal-perfluorohexane as the test fluid and will operate between pressures of 60 to 244 Pa. and temperatures of 35 to 60 C. Both sets of experimental heaters are highly instrumented. Pressure and bulk fluid temperature measurements will be made with standard rate video. A high speed video system will be used to visualize the boiling process through the bottom of the MABE heater arrays. The BXF is currently scheduled to fly on Utilization Flight-13A.1 to the ISS with facility integration into the MSG and operation during Increment 15

  6. An Investigation of Graduate Scientists' Understandings of Evaporation and Boiling.

    ERIC Educational Resources Information Center

    Goodwin, Alan; Orlik, Yuri

    2000-01-01

    Uses a video presentation of six situations relating to the evaporation and boiling of liquids and the escape of dissolved gases from solution and investigates graduate scientists' understanding of the concepts of boiling and evaporation. (Author/YDS)

  7. New Departure from Nucleate Boiling model relying on first principle energy balance at the boiling surface

    NASA Astrophysics Data System (ADS)

    Demarly, Etienne; Baglietto, Emilio

    2017-11-01

    Predictions of Departure from Nucleate Boiling have been a longstanding challenge when designing heat exchangers such as boilers or nuclear reactors. Many mechanistic models have been postulated over more than 50 years in order to explain this phenomenon but none is able to predict accurately the conditions which trigger the sudden change of heat transfer mode. This work aims at demonstrating the pertinence of a new approach for detecting DNB by leveraging recent experimental insights. The new model proposed departs from all the previous models by making the DNB inception come from an energy balance instability at the heating surface rather than a hydrodynamic instability of the bubbly layer above the surface (Zuber, 1959). The main idea is to modulate the amount of heat flux being exchanged via the nucleate boiling mechanism by the wetted area fraction on the surface, thus allowing a completely automatic trigger of DNB that doesn't require any parameter prescription. This approach is implemented as a surrogate model in MATLAB in order to validate the principles of the model in a simple and controlled geometry. Good agreement is found with the experimental data leveraged from the MIT Flow Boiling at various flow regimes. Consortium for Advanced Simulation of Light Water Reactors (CASL).

  8. Gravity and Heater Size Effects on Pool Boiling Heat Transfer

    NASA Technical Reports Server (NTRS)

    Kim, Jungho; Raj, Rishi

    2014-01-01

    The current work is based on observations of boiling heat transfer over a continuous range of gravity levels between 0g to 1.8g and varying heater sizes with a fluorinert as the test liquid (FC-72/n-perfluorohexane). Variable gravity pool boiling heat transfer measurements over a wide range of gravity levels were made during parabolic flight campaigns as well as onboard the International Space Station. For large heaters and-or higher gravity conditions, buoyancy dominated boiling and heat transfer results were heater size independent. The power law coefficient for gravity in the heat transfer equation was found to be a function of wall temperature under these conditions. Under low gravity conditions and-or for smaller heaters, surface tension forces dominated and heat transfer results were heater size dependent. A pool boiling regime map differentiating buoyancy and surface tension dominated regimes was developed along with a unified framework that allowed for scaling of pool boiling over a wide range of gravity levels and heater sizes. The scaling laws developed in this study are expected to allow performance quantification of phase change based technologies under variable gravity environments eventually leading to their implementation in space based applications.

  9. A numerical investigation of the effect of surface wettability on the boiling curve.

    PubMed

    Hsu, Hua-Yi; Lin, Ming-Chieh; Popovic, Bridget; Lin, Chii-Ruey; Patankar, Neelesh A

    2017-01-01

    Surface wettability is recognized as playing an important role in pool boiling and the corresponding heat transfer curve. In this work, a systematic study of pool boiling heat transfer on smooth surfaces of varying wettability (contact angle range of 5° - 180°) has been conducted and reported. Based on numerical simulations, boiling curves are calculated and boiling dynamics in each regime are studied using a volume-of-fluid method with contact angle model. The calculated trends in critical heat flux and Leidenfrost point as functions of surface wettability are obtained and compared with prior experimental and theoretical predictions, giving good agreement. For the first time, the effect of contact angle on the complete boiling curve is shown. It is demonstrated that the simulation methodology can be used for studying pool boiling and related dynamics and providing more physical insights.

  10. A numerical investigation of the effect of surface wettability on the boiling curve

    PubMed Central

    Lin, Ming-Chieh; Popovic, Bridget; Lin, Chii-Ruey; Patankar, Neelesh A.

    2017-01-01

    Surface wettability is recognized as playing an important role in pool boiling and the corresponding heat transfer curve. In this work, a systematic study of pool boiling heat transfer on smooth surfaces of varying wettability (contact angle range of 5° − 180°) has been conducted and reported. Based on numerical simulations, boiling curves are calculated and boiling dynamics in each regime are studied using a volume-of-fluid method with contact angle model. The calculated trends in critical heat flux and Leidenfrost point as functions of surface wettability are obtained and compared with prior experimental and theoretical predictions, giving good agreement. For the first time, the effect of contact angle on the complete boiling curve is shown. It is demonstrated that the simulation methodology can be used for studying pool boiling and related dynamics and providing more physical insights. PMID:29125847

  11. Orthogonal typing methods identify genetic diversity among Belgian Campylobacter jejuni strains isolated over a decade from poultry and cases of sporadic human illness.

    PubMed

    Elhadidy, Mohamed; Arguello, Hector; Álvarez-Ordóñez, Avelino; Miller, William G; Duarte, Alexandra; Martiny, Delphine; Hallin, Marie; Vandenberg, Olivier; Dierick, Katelijne; Botteldoorn, Nadine

    2018-06-20

    Campylobacter jejuni is a zoonotic pathogen commonly associated with human gastroenteritis. Retail poultry meat is a major food-related transmission source of C. jejuni to humans. The present study investigated the genetic diversity, clonal relationship, and strain risk-analysis of 403 representative C. jejuni isolates from chicken broilers (n = 204) and sporadic cases of human diarrhea (n = 199) over a decade (2006-2015) in Belgium, using multilocus sequence typing (MLST), PCR binary typing (P-BIT), and identification of lipooligosaccharide (LOS) biosynthesis locus classes. A total of 123 distinct sequence types (STs), clustered in 28 clonal complexes (CCs) were assigned, including ten novel sequence types that were not previously documented in the international database. Sequence types ST-48, ST-21, ST-50, ST-45, ST-464, ST-2274, ST-572, ST-19, ST-257 and ST-42 were the most prevalent. Clonal complex 21 was the main clonal complex in isolates from humans and chickens. Among observed STs, a total of 35 STs that represent 72.2% (291/403) of the isolates were identified in both chicken and human isolates confirming considerable epidemiological relatedness; these 35 STs also clustered together in the most prevalent CCs. A majority of the isolates harbored sialylated LOS loci associated with potential neuropathic outcomes in humans. Although the concordance between MLST and P-BIT, determined by the adjusted Rand and Wallace coefficients, showed low congruence between both typing methods. The discriminatory power of P-BIT and MLST was similar, with Simpson's diversity indexes of 0.978 and 0.975, respectively. Furthermore, P-BIT could provide additional epidemiological information that would provide further insights regarding the potential association to human health from each strain. In addition, certain clones could be linked to specific clinical symptoms. Indeed, LOS class E was associated with less severe infections. Moreover, ST-572 was significantly

  12. A high-fidelity approach towards simulation of pool boiling

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Yazdani, Miad; Radcliff, Thomas; Soteriou, Marios

    2016-01-15

    A novel numerical approach is developed to simulate the multiscale problem of pool-boiling phase change. The particular focus is to develop a simulation technique that is capable of predicting the heat transfer and hydrodynamic characteristics of nucleate boiling and the transition to critical heat flux on surfaces of arbitrary shape and roughness distribution addressing a critical need to design enhanced boiling heat transfer surfaces. The macro-scale of the phase change and bubble dynamics is addressed through employing off-the-shelf Computational Fluid Dynamics (CFD) methods for interface tracking and interphase mass and energy transfer. The micro-scale of the microlayer, which forms atmore » early stage of bubble nucleation near the wall, is resolved through asymptotic approximation of the thin-film theory which provides a closed-form solution for the distribution of the micro-layer and its influence on the evaporation process. In addition, the sub-grid surface roughness is represented stochastically through probabilistic density functions and its role in bubble nucleation and growth is then represented based on the thermodynamics of nucleation process. This combination of deterministic CFD, local approximation, and stochastic representation allows the simulation of pool boiling on any surface with known roughness and enhancement characteristics. The numerical model is validated for dynamics and hydrothermal characteristics of a single nucleated bubble on a flat surface against available literature data. In addition, the prediction of pool-boiling heat transfer coefficient is verified against experimental measurements as well as reputable correlations for various roughness distributions and different surface orientations. Finally, the model is employed to demonstrate pool-boiling phenomenon on enhanced structures with reentrance cavities and to explore the effect of enhancement feature design on thermal and hydrodynamic characteristics of these surfaces.« less

  13. A study of forced convection boiling under reduced gravity

    NASA Technical Reports Server (NTRS)

    Merte, Herman, Jr.

    1992-01-01

    This report presents the results of activities conducted over the period 1/2/85-12/31/90, in which the study of forced convection boiling under reduced gravity was initiated. The study seeks to improve the understanding of the basic processes that constitute forced convection boiling by removing the buoyancy effects which may mask other phenomena. Specific objectives may also be expressed in terms of the following questions: (1) what effects, if any, will the removal of body forces to the lowest possible levels have on the forced convection boiling heat transfer processes in well-defined and meaningful circumstances? (this includes those effects and processes associated with the nucleation or onset of boiling during the transient increase in heater surface temperature, as well as the heat transfer and vapor bubble behaviors with established or steady-state conditions); and (2) if such effects are present, what are the boundaries of the relevant parameters such as heat flux, heater surface superheat, fluid velocity, bulk subcooling, and geometric/orientation relationships within which such effects will be produced?

  14. Boiling process modelling peculiarities analysis of the vacuum boiler

    NASA Astrophysics Data System (ADS)

    Slobodina, E. N.; Mikhailov, A. G.

    2017-06-01

    The analysis of the low and medium powered boiler equipment development was carried out, boiler units possible development directions with the purpose of energy efficiency improvement were identified. Engineering studies for the vacuum boilers applying are represented. Vacuum boiler heat-exchange processes where boiling water is the working body are considered. Heat-exchange intensification method under boiling at the maximum heat- transfer coefficient is examined. As a result of the conducted calculation studies, heat-transfer coefficients variation curves depending on the pressure, calculated through the analytical and numerical methodologies were obtained. The conclusion about the possibility of numerical computing method application through RPI ANSYS CFX for the boiling process description in boiler vacuum volume was given.

  15. Purification, characterization and sequence analysis of Omp50,a new porin isolated from Campylobacter jejuni.

    PubMed Central

    Bolla, J M; Dé, E; Dorez, A; Pagès, J M

    2000-01-01

    A novel pore-forming protein identified in Campylobacter was purified by ion-exchange chromatography and named Omp50 according to both its molecular mass and its outer membrane localization. We observed a pore-forming ability of Omp50 after re-incorporation into artificial membranes. The protein induced cation-selective channels with major conductance values of 50-60 pS in 1 M NaCl. N-terminal sequencing allowed us to identify the predicted coding sequence Cj1170c from the Campylobacter jejuni genome database as the corresponding gene in the NCTC 11168 genome sequence. The gene, designated omp50, consists of a 1425 bp open reading frame encoding a deduced 453-amino acid protein with a calculated pI of 5.81 and a molecular mass of 51169.2 Da. The protein possessed a 20-amino acid leader sequence. No significant similarity was found between Omp50 and porin protein sequences already determined. Moreover, the protein showed only weak sequence identity with the major outer-membrane protein (MOMP) of Campylobacter, correlating with the absence of antigenic cross-reactivity between these two proteins. Omp50 is expressed in C. jejuni and Campylobacter lari but not in Campylobacter coli. The gene, however, was detected in all three species by PCR. According to its conformation and functional properties, the protein would belong to the family of outer-membrane monomeric porins. PMID:11104668

  16. 17. RW Meyer Sugar Mill: 18761889. Boiling House, 1878. View: ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    17. RW Meyer Sugar Mill: 1876-1889. Boiling House, 1878. View: Southwest corner of boiling house. The amimal-powered cane mill is located in the undergrowth in the right foreground, - R. W. Meyer Sugar Mill, State Route 47, Kualapuu, Maui County, HI

  17. Extended hydrodynamic theory of the peak and minimum pool boiling heat fluxes

    NASA Technical Reports Server (NTRS)

    Linehard, J. H.; Dhir, V. K.

    1973-01-01

    The hydrodynamic theory of the extreme pool boiling heat fluxes is expanded to embrace a variety of problems that have not previously been analyzed. These problems include the prediction of the peak heat flux on a variety of finite heaters, the influence of viscosity on the Taylor and Helmoltz instability mechanisms with application to film boiling and to the peak heat flux in viscous liquids, the formalization of the analogy between high-current-density electrolysis and boiling, and the description of boiling in the low-gravity limit. The predictions are verified with a large number of new data.

  18. Enhanced Boiling on Micro-Configured Composite Surfaces Under Microgravity Conditions

    NASA Technical Reports Server (NTRS)

    Zhang, Nengli; Chai, An-Ti

    1999-01-01

    In order to accommodate the growing thermal management needs of future space platforms, several two-phase active thermal control systems (ATCSs) have evolved and were included in the designs of space stations. Compared to the pumped single-phase liquid loops used in the conventional Space Transportation System and Spacelab, ATCSs offer significant benefits that may be realized by adopting a two-phase fluid-loop system. Alternately, dynamic power systems (DPSs), based on the Rankine cycle, seem inevitably to be required to supply the electrical power requirements of expanding space activities. Boiling heat transfer is one of the key technologies for both ATCSs and DPSs. Nucleate boiling near critical heat flux (CHF) can transport very large thermal loads with much smaller device size and much lower pumping power. However, boiling performance deteriorates in a reduced gravity environment and operation in the CHF regime is precarious because any slight overload will cause the heat transfer to suddenly move to the film boiling regime, which in turn, will result in burnout of the heat transfer surfaces. New materials, such as micro-configured metal-graphite composites, can provide a solution for boiling enhancement. It has been shown experimentally that this type of material manifests outstanding boiling heat transfer performance and their CHF is also extended to higher values. Due to the high thermal conductivity of graphite fiber (up to 1,200 W/m-K in the fiber direction), the composite surfaces are non-isothermal during the boiling process. The composite surfaces are believed to have a much wider safe operating region (a more uniform boiling curve in the CHF regime) because non-isothermal surfaces have been found to be less sensitive to variations of wall superheat in the CHF regime. The thermocapillary forces formed by the temperature difference between the fiber tips and the metal matrix play a more important role than the buoyancy in the bubble detachment, for the

  19. Turning bubbles on and off during boiling using charged surfactants

    PubMed Central

    Cho, H. Jeremy; Mizerak, Jordan P.; Wang, Evelyn N.

    2015-01-01

    Boiling—a process that has powered industries since the steam age—is governed by bubble formation. State-of-the-art boiling surfaces often increase bubble nucleation via roughness and/or wettability modification to increase performance. However, without active in situ control of bubbles, temperature or steam generation cannot be adjusted for a given heat input. Here we report the ability to turn bubbles ‘on and off' independent of heat input during boiling both temporally and spatially via molecular manipulation of the boiling surface. As a result, we can rapidly and reversibly alter heat transfer performance up to an order of magnitude. Our experiments show that this active control is achieved by electrostatically adsorbing and desorbing charged surfactants to alter the wettability of the surface, thereby affecting nucleation. This approach can improve performance and flexibility in existing boiling technologies as well as enable emerging or unprecedented energy applications. PMID:26486275

  20. Zero Boil Off Cryogen Storage for Future Launchers

    NASA Technical Reports Server (NTRS)

    Valentian, D.; Plachta, D.; Kittel, P.; Hastings, L. J.; Salerno, Louis J.; Arnold, James O. (Technical Monitor)

    2001-01-01

    Zero boil off (ZBO) cryogen storage using both cryocoolers and passive insulation technologies will enable long-term exploration missions by allowing designers to optimize tankage without the need for excess cryogen storage to account for boil off. Studies of ZBO (zero boil off) have been on-going in the USA for several years. More recently, a review of the needs of advanced space propulsion took place in Europe. This showed the interest of the European community in cryogenic propulsion for planetary missions as well as the use of liquid hydrogen for large power electric propulsion (manned Mars missions). Although natural boiling could be acceptable for single leg missions, passive insulation techniques yield roughly a I% per month cryogen loss and this would not be cost effective for robotic planetary missions involving storage times greater than one year. To make economic sense, long-term exploration missions require lower tank capacity and longer storage times. Recent advances in cryocooler technology, resulting in vast improvements in both cooler efficiency and reliability, make ZBO is a clear choice for planetary exploration missions. Other, more near term applications of ZBO include boil-off reduction or elimination applied to first and upper stages of future earth-to-orbit (ETO) launchers. This would extend launch windows and reduce infrastructure costs. Successors to vehicles like Ariane 5 could greatly benefit by implementing ZBO. Zero Boil Off will only be successful in ETO launcher applications if it makes economic sense to implement. The energy cost is only a fraction of the total cost of buying liquid cryogen, the rest being transportation and other overhead. Because of this, higher boiling point cryogens will benefit more from on-board liquefaction, thus reducing the infrastructure costs. Since hydrogen requires a liquefier with at least a 17% efficiency just to break even from a cost standpoint, one approach for implementing ZBO in upper stages would

  1. Drug resistance in Campylobacter jejuni, C coli, and C lari isolated from humans in north west England and Wales, 1997.

    PubMed Central

    Thwaites, R T; Frost, J A

    1999-01-01

    AIMS: To test the sensitivity of strains of Campylobacter species isolated from humans in England and Wales against a range of antimicrobial agents for the purpose of monitoring therapeutic efficacy and as an epidemiological marker. METHODS: An agar dilution breakpoint technique was used to screen isolates against ampicillin, chloramphenicol, gentamicin, kanamycin, neomycin, tetracycline, nalidixic acid, ciprofloxacin, and erythromycin. Minimal inhibitory concentrations (MIC) were also determined for a sample of quinolone resistant strains. RESULTS: Approximately 50% of strains tested were resistant to at least one drug. Strains which were resistant to four or more of the drugs tested were classified as multiresistant; this occurred in 11.3% of C jejuni, 19.9% of C coli, and 63.6% of C lari. Resistance to erythromycin occurred in 1.0% of C jejuni and 12.8% of C coli. Resistance to quinolones occurred in 12% of strains, with a ciprofloxacin MIC of > 8 mg/l and a nalidixic acid MIC of > 256 mg/l; a further 4% of strains had intermediate resistance with a ciprofloxacin MIC of between 0.5 and 2 mg/l (fully sensitive strains, 0.25 mg/l or less) and a nalidixic acid MIC of between 32 and 64 mg/l (fully sensitive strains, 8 mg/l or less). CONCLUSIONS: Resistance to quinolones in campylobacters from human infection may relate to clinical overuse or use of fluoroquinolones in animal husbandry. Both veterinary and clinical use should be reconsidered and fluoroquinolone drugs used only as a treatment for serious infections requiring hospital admission. Erythromycin resistance is still rare in C jejuni but much more common in C coli. PMID:10690169

  2. Boiling regimes of impacting drops on a heated substrate under reduced pressure

    NASA Astrophysics Data System (ADS)

    van Limbeek, Michiel A. J.; Hoefnagels, Paul B. J.; Shirota, Minori; Sun, Chao; Lohse, Detlef

    2018-05-01

    We experimentally investigate the boiling behavior of impacting ethanol drops on a heated smooth sapphire substrate at pressures ranging from P =0.13 bar to atmospheric pressure. We employ frustrated total internal reflection imaging to study the wetting dynamics of the contact between the drop and the substrate. The spreading drop can be in full contact (contact boiling), it can partially touch (transition boiling), or the drop can be fully levitated (Leidenfrost boiling). We show that the temperature of the boundary between contact and transition boiling shows at most a weak dependence on the impact velocity, but a significant decrease with decreasing ambient gas pressure. A striking correspondence is found between the temperature of this boundary and the static Leidenfrost temperature for all pressures. We therefore conclude that both phenomena share the same mechanism and are dominated by the dynamics taking place at the contact line. On the other hand, the boundary between transition boiling and Leidenfrost boiling, i.e., the dynamic Leidenfrost temperature, increases for increasing impact velocity for all ambient gas pressures. Moreover, the dynamic Leidenfrost temperature coincides for pressures between P =0.13 and 0.54 bar, whereas for atmospheric pressure the dynamic Leidenfrost temperature is slightly elevated. This indicates that the dynamic Leidenfrost temperature is at most weakly dependent on the enhanced evaporation by the lower saturation temperature of the liquid.

  3. An Analytical Approach for Relating Boiling Points of Monofunctional Organic Compounds to Intermolecular Forces

    ERIC Educational Resources Information Center

    Struyf, Jef

    2011-01-01

    The boiling point of a monofunctional organic compound is expressed as the sum of two parts: a contribution to the boiling point due to the R group and a contribution due to the functional group. The boiling point in absolute temperature of the corresponding RH hydrocarbon is chosen for the contribution to the boiling point of the R group and is a…

  4. Campylobacter jejuni dsb gene expression is regulated by iron in a Fur-dependent manner and by a translational coupling mechanism

    PubMed Central

    2011-01-01

    Background Many bacterial extracytoplasmic proteins are stabilized by intramolecular disulfide bridges that are formed post-translationally between their cysteine residues. This protein modification plays an important role in bacterial pathogenesis, and is facilitated by the Dsb (disulfide bond) family of the redox proteins. These proteins function in two parallel pathways in the periplasmic space: an oxidation pathway and an isomerization pathway. The Dsb oxidative pathway in Campylobacter jejuni is more complex than the one in the laboratory E. coli K-12 strain. Results In the C. jejuni 81-176 genome, the dsb genes of the oxidative pathway are arranged in three transcriptional units: dsbA2-dsbB-astA, dsbA1 and dba-dsbI. Their transcription responds to an environmental stimulus - iron availability - and is regulated in a Fur-dependent manner. Fur involvement in dsb gene regulation was proven by a reporter gene study in a C. jejuni wild type strain and its isogenic fur mutant. An electrophoretic mobility shift assay (EMSA) confirmed that analyzed genes are members of the Fur regulon but each of them is regulated by a disparate mechanism, and both the iron-free and the iron-complexed Fur are able to bind in vitro to the C. jejuni promoter regions. This study led to identification of a new iron- and Fur-regulated promoter that drives dsbA1 gene expression in an indirect way. Moreover, the present work documents that synthesis of DsbI oxidoreductase is controlled by the mechanism of translational coupling. The importance of a secondary dba-dsbI mRNA structure for dsbI mRNA translation was verified by estimating individual dsbI gene expression from its own promoter. Conclusions The present work shows that iron concentration is a significant factor in dsb gene transcription. These results support the concept that iron concentration - also through its influence on dsb gene expression - might control the abundance of extracytoplasmic proteins during different stages of

  5. Zero Boil-Off System Testing

    NASA Technical Reports Server (NTRS)

    Plachta, David W.; Johnson, Wesley L.; Feller, Jeffrey R.

    2015-01-01

    Cryogenic propellants such as liquid hydrogen (LH2) and liquid oxygen (LO2) are a part of NASA's future space exploration due to their high specific impulse for rocket motors of upper stages suitable for transporting 10s to 100s of metric tons of payload mass to destinations outside of low earth orbit and for their return. However, the low storage temperatures of LH2 and LO2 cause substantial boil-off losses for missions with durations greater than several months. These losses can be eliminated by incorporating high performance cryocooler technology to intercept heat load to the propellant tanks and modulating the cryocooler to control tank pressure. The active thermal control technology being developed by NASA is the reverse turbo-Brayton cycle cryocooler and its integration to the propellant tank through a distributed cooling tubing network coupled to the tank wall. This configuration was recently tested at NASA Glenn Research Center, in a vacuum chamber and cryoshroud that simulated the essential thermal aspects of low Earth orbit, its vacuum and temperature. Testing consisted of three passive tests with the active cryocooler system off, and 7 active tests, with the cryocooler powered up. The test matrix included zero boil-off tests performed at 90 full and 25 full, and several demonstrations at excess cooling capacity and reduced cooling capacity. From this, the tank pressure response with varied cryocooler power inputs was determined. This test series established that the active cooling system integrated with the propellant tank eliminated boil-off and robustly controlled tank pressure.

  6. Pool boiling of water on nano-structured micro wires at sub-atmospheric conditions

    NASA Astrophysics Data System (ADS)

    Arya, Mahendra; Khandekar, Sameer; Pratap, Dheeraj; Ramakrishna, S. Anantha

    2016-09-01

    Past decades have seen active research in enhancement of boiling heat transfer by surface modifications. Favorable surface modifications are expected to enhance boiling efficiency. Several interrelated mechanisms such as capillarity, surface energy alteration, wettability, cavity geometry, wetting transitions, geometrical features of surface morphology, etc., are responsible for change in the boiling behavior of modified surfaces. Not much work is available on pool boiling at low pressures on microscale/nanoscale geometries; low pressure boiling is attractive in many applications wherein low operating temperatures are desired for a particular working fluid. In this background, an experimental setup was designed and developed to investigate the pool boiling performance of water on (a) plain aluminum micro wire (99.999 % pure) and, (b) nano-porous alumina structured aluminum micro wire, both having diameter of 250 µm, under sub-atmospheric pressure. Nano-structuring on the plain wire surface was achieved via anodization. Two samples, A and B of anodized wires, differing by the degree of anodization were tested. The heater length scale (wire diameter) was much smaller than the capillary length scale. Pool boiling characteristics of water were investigated at three different sub-atmospheric pressures of 73, 123 and 199 mbar (corresponding to T sat = 40, 50 and 60 °C). First, the boiling characteristics of plain wire were measured. It was noticed that at sub-atmospheric pressures, boiling heat transfer performance for plain wire was quite low due to the increased bubble sizes and low nucleation site density. Subsequently, boiling performance of nano-structured wires (both Sample A and Sample B) was compared with plain wire and it was noted that boiling heat transfer for the former was considerably enhanced as compared to the plain wire. This enhancement is attributed to increased nucleation site density, change in wettability and possibly due to enhanced pore scale

  7. Length Scale and Gravity Effects on Microgravity Boiling Heat Transfer

    NASA Technical Reports Server (NTRS)

    Kim, Jungho; McQuillen, John; Balombin, Joe

    2002-01-01

    Boiling is a complex phenomenon where hydrodynamics, heat transfer, mass transfer, and interfacial phenomena are tightly interwoven. An understanding of boiling and critical heat flux in microgravity environments is of importance to space based hardware and processes such as heat exchange, cryogenic fuel storage and transportation, electronic cooling, and material processing due to the large amounts of heat that can be removed with relatively little increase in temperature. Although research in this area has been performed in the past four decades, the mechanisms by which heat is removed from surfaces in microgravity are still unclear. In earth gravity, buoyancy is an important parameter that affects boiling heat transfer through the rate at which bubbles are removed from the surface. A simple model describing the bubble departure size based on a quasistatic force balance between buoyancy and surface tension is given by the Fritz [I] relation: Bo(exp 1/2) = 0.0208 theta where Bo is the ratio between buoyancy and surface tension forces. For small, rapidly growing bubbles, inertia associated with the induced liquid motion can also cause bubble departure. In microgravity, the magnitude of effects related to natural convection and buoyancy are small and physical mechanisms normally masked by natural convection in earth gravity such as Marangoni convection can substantially influence the boiling and vapor bubble dynamics. CHF (critical heat transfer) is also substantially affected by microgravity. In 1 g environments, Bo has been used as a correlating parameter for CHF. Zuber's CHF model for an infinite horizontal surface assumes that vapor columns formed by the merger of bubbles become unstable due to a Helmholtz instability blocking the supply of liquid to the surface. The jets are spaced lambda(sub D) apart, where lambda(sub D) = 2pi square root of 3[(sigma)/(g(rho(sub l) - rho(sub v)](exp 1/2) = 2pi square root of 3 L Bo(exp -1/2) = square root of 3 lambda(sub c

  8. QSPR using MOLGEN-QSPR: the challenge of fluoroalkane boiling points.

    PubMed

    Rücker, Christoph; Meringer, Markus; Kerber, Adalbert

    2005-01-01

    By means of the new software MOLGEN-QSPR, a multilinear regression model for the boiling points of lower fluoroalkanes is established. The model is based exclusively on simple descriptors derived directly from molecular structure and nevertheless describes a broader set of data more precisely than previous attempts that used either more demanding (quantum chemical) descriptors or more demanding (nonlinear) statistical methods such as neural networks. The model's internal consistency was confirmed by leave-one-out cross-validation. The model was used to predict all unknown boiling points of fluorobutanes, and the quality of predictions was estimated by means of comparison with boiling point predictions for fluoropentanes.

  9. Teaching Structure-Property Relationships: Investigating Molecular Structure and Boiling Point

    ERIC Educational Resources Information Center

    Murphy, Peter M.

    2007-01-01

    A concise, well-organized table of the boiling points of 392 organic compounds has facilitated inquiry-based instruction in multiple scientific principles. Many individual or group learning activities can be derived from the tabulated data of molecular structure and boiling point based on the instructor's education objectives and the students'…

  10. On mechanism of explosive boiling in nanosecond regime

    NASA Astrophysics Data System (ADS)

    Çelen, Serap

    2016-06-01

    Today laser-based machining is used to manufacture vital parts for biomedical, aviation and aerospace industries. The aim of the paper is to report theoretical, numerical and experimental investigations of explosive boiling under nanosecond pulsed ytterbium fiber laser irradiation. Experiments were performed in an effective peak power density range between 1397 and 1450 MW/cm2 on pure titanium specimens. The threshold laser fluence for phase explosion, the pressure and temperature at the target surface and the velocity of the expulsed material were reported. A narrow transition zone was realized between the normal vaporization and phase explosion fields. The proof of heterogeneous boiling was given with detailed micrographs. A novel thermal model was proposed for laser-induced splashing at high fluences. Packaging factor and scattering arc radius terms were proposed to state the level of the melt ejection process. Results of the present investigation explain the explosive boiling during high-power laser interaction with metal.

  11. An Experimental Study of Boiling in Reduced and Zero Gravity Fields

    NASA Technical Reports Server (NTRS)

    Usiskin, C. M.; Siegel, R.

    1961-01-01

    A pool boiling apparatus was mounted on a counterweighted platform which could be dropped a distance of nine feet. By varying the size of the counterweight, the effective gravity field on the equipment was adjusted between zero and unity. A study of boiling burnout in water indicated that a variation in the critical heat flux according to the one quarter power of gravity was reasonable. A consideration of the transient burnout process was necessary in order to properly interpret the data. A photographic study of nucleate boiling showed how the velocity of freely rising vapor bubbles decreased as gravity was reduced. The bubble diameters at the time of breakoff from the heated surface were found to vary inversely as gravity to the 1/3.5 power. Motion pictures were taken to illustrate both nucleate and film boiling in the low gravity range.

  12. Characterization of Lipooligosaccharide-Biosynthetic Loci of Campylobacter jejuni Reveals New Lipooligosaccharide Classes: Evidence of Mosaic Organizations▿ †

    PubMed Central

    Parker, Craig T.; Gilbert, Michel; Yuki, Nobuhiro; Endtz, Hubert P.; Mandrell, Robert E.

    2008-01-01

    The lipooligosaccharide (LOS) biosynthesis region is one of the more variable genomic regions between strains of Campylobacter jejuni. Indeed, eight classes of LOS biosynthesis loci have been established previously based on gene content and organization. In this study, we characterize additional classes of LOS biosynthesis loci and analyze various mechanisms that result in changes to LOS structures. To gain further insights into the genomic diversity of C. jejuni LOS biosynthesis region, we sequenced the LOS biosynthesis loci of 15 strains that possessed gene content that was distinct from the eight classes. This analysis identified 11 new classes of LOS loci that exhibited examples of deletions and insertions of genes and cassettes of genes found in other LOS classes or capsular biosynthesis loci leading to mosaic LOS loci. The sequence analysis also revealed both missense mutations leading to “allelic” glycosyltransferases and phase-variable and non-phase-variable gene inactivation by the deletion or insertion of bases. Specifically, we demonstrated that gene inactivation is an important mechanism for altering the LOS structures of strains possessing the same class of LOS biosynthesis locus. Together, these observations suggest that LOS biosynthesis region is a hotspot for genetic exchange and variability, often leading to changes in the LOS produced. PMID:18556784

  13. A Study of Nucleate Boiling with Forced Convection in Microgravity

    NASA Technical Reports Server (NTRS)

    Merte, Herman, Jr.

    1999-01-01

    The ultimate objective of basic studies of flow boiling in microgravity is to improve the understanding of the processes involved, as manifested by the ability to predict its behavior. This is not yet the case for boiling heat transfer even in earth gravity, despite the considerable research activity over the past 30 years. The elements that constitute the nucleate boiling process - nucleation, growth, motion, and collapse of the vapor bubbles (if the bulk liquid is subcooled) - are common to both pool and flow boiling. It is well known that the imposition of bulk liquid motion affects the vapor bubble behavior relative to pool boiling, but does not appear to significantly influence the heat transfer. Indeed, it has been recommended in the past that empirical correlations or experimental data of pool boiling be used for design purposes with forced convection nucleate boiling. It is anticipated that such will most certainly not be possible for boiling in microgravity, based on observations made with pool boiling in microgravity. In earth gravity buoyancy will act to remove the vapor bubbles from the vicinity of the heater surface regardless of how much the imposed bulk velocity is reduced, depending, of course, on the geometry of the system. Vapor bubbles have been observed to dramatically increase in size in pool boiling in microgravity, and the heat flux at which dryout took place was reduced considerably below what is generally termed the critical heat flux (CHF) in earth gravity, depending on the bulk liquid subcooling. However, at heat flux levels below dryout, the nucleate pool boiling process was enhanced considerably over that in earth gravity, in spite of the large vapor bubbles formed in microgravity and perhaps as a consequence. These large vapor bubbles tended to remain in the vicinity of the heater surface, and the enhanced heat transfer appeared to be associated with the presence of what variously has been referred to as a liquid microlayer between the

  14. Fundamental Boiling and RP-1 Freezing Experiments

    NASA Technical Reports Server (NTRS)

    Goode, Brian; Turner, Larry D. (Technical Monitor)

    2001-01-01

    This paper describes results from experiments performed to help understand certain aspects of the MC-1 engine prestart thermal conditioning procedure. The procedure was constrained by the fact that the engine must chill long enough to get quality LOX at the LOX pump inlet but must be short enough to prevent freezing of RP-1 in the fuel pump. A chill test of an MC-1 LOX impeller was performed in LN2 to obtain data on film boiling, transition boiling and impeller temperature histories. The transition boiling data was important to the chill time so a subsequent experiment was performed chilling simple steel plates in LOX to obtain similar data for LOX. To address the fuel freezing concern, two experiments were performed. First, fuel was frozen in a tray and its physical characteristics were observed and temperatures of the fuel were measured. The result was physical characteristics as a function of temperature. Second was an attempt to measure the frozen thickness of RP-1 on a cold wall submerged in warm RP-1 and to develop a method for calculating that thickness for other conditions.

  15. A Novel Role of Three Dimensional Graphene Foam to Prevent Heater Failure during Boiling

    PubMed Central

    Ahn, Ho Seon; Kim, Ji Min; Park, Chibeom; Jang, Ji-Wook; Lee, Jae Sung; Kim, Hyungdae; Kaviany, Massoud; Kim, Moo Hwan

    2013-01-01

    We report a novel boiling heat transfer (NBHT) in reduced graphene oxide (RGO) suspended in water (RGO colloid) near critical heat flux (CHF), which is traditionally the dangerous limitation of nucleate boiling heat transfer because of heater failure. When the heat flux reaches the maximum value (CHF) in RGO colloid pool boiling, the wall temperature increases gradually and slowly with an almost constant heat flux, contrary to the rapid wall temperature increase found during water pool boiling. The gained time by NBHT would provide the safer margin of the heat transfer and the amazing impact on the thermal system as the first report of graphene application. In addition, the CHF and boiling heat transfer performance also increase. This novel boiling phenomenon can effectively prevent heater failure because of the role played by the self-assembled three-dimensional foam-like graphene network (SFG). PMID:23743619

  16. Zero boil-off system testing

    NASA Astrophysics Data System (ADS)

    Plachta, D. W.; Johnson, W. L.; Feller, J. R.

    2016-03-01

    Cryogenic propellants such as liquid hydrogen (LH2) and liquid oxygen (LO2) are a part of NASA's future space exploration plans due to their high specific impulse for rocket motors of upper stages. However, the low storage temperatures of LH2 and LO2 cause substantial boil-off losses for long duration missions. These losses can be eliminated by incorporating high performance cryocooler technology to intercept heat load to the propellant tanks and modulating the cryocooler temperature to control tank pressure. The technology being developed by NASA is the reverse turbo-Brayton cycle cryocooler and its integration to the propellant tank through a distributed cooling tubing network coupled to the tank wall. This configuration was recently tested at NASA Glenn Research Center in a vacuum chamber and cryoshroud that simulated the essential thermal aspects of low Earth orbit, its vacuum and temperature. This test series established that the active cooling system integrated with the propellant tank eliminated boil-off and robustly controlled tank pressure.

  17. Zero Boil-Off System Testing

    NASA Technical Reports Server (NTRS)

    Plachta, D. W.; Johnson, W. L.; Feller, J. R.

    2015-01-01

    Cryogenic propellants such as liquid hydrogen (LH2) and liquid oxygen (LO2) are a part of NASA's future space exploration plans due to their high specific impulse for rocket motors of upper stages. However, the low storage temperatures of LH2 and LO2 cause substantial boil-off losses for long duration missions. These losses can be eliminated by incorporating high performance cryocooler technology to intercept heat load to the propellant tanks and modulating the cryocooler temperature to control tank pressure. The technology being developed by NASA is the reverse turbo-Brayton cycle cryocooler and its integration to the propellant tank through a distributed cooling tubing network coupled to the tank wall. This configuration was recently tested at NASA Glenn Research Center in a vacuum chamber and cryoshroud that simulated the essential thermal aspects of low Earth orbit, its vacuum and temperature. This test series established that the active cooling system integrated with the propellant tank eliminated boil-off and robustly controlled tank pressure.

  18. BOILING SLURRY REACTOR AND METHOD FO CONTROL

    DOEpatents

    Petrick, M.; Marchaterre, J.F.

    1963-05-01

    The control of a boiling slurry nuclear reactor is described. The reactor consists of a vertical tube having an enlarged portion, a steam drum at the top of the vertical tube, and at least one downcomer connecting the steam drum and the bottom of the vertical tube, the reactor being filled with a slurry of fissionabie material in water of such concentration that the enlarged portion of the vertical tube contains a critical mass. The slurry boils in the vertical tube and circulates upwardly therein and downwardly in the downcomer. To control the reactor by controlling the circulation of the slurry, a gas is introduced into the downcomer. (AEC)

  19. Nucleate pool boiling in the long duration low gravity environment of the space shuttle

    NASA Technical Reports Server (NTRS)

    Hasan, M. M.; Lin, C. S.; Knoll, R. H.; Bentz, M. D.; Meserole, J. S.

    1993-01-01

    The results are presented of an experimental study of nucleate pool boiling performed in the low gravity environment of the space shuttle. Photographic observations of pool boiling in Freon 113 were obtained during the 'Tank Pressure Control Experiment', flown on the Space Transportation System STS-43 in August 1991. Nucleate boiling data from large (relative to bubble size) flat heating surfaces (0.1046 by 0.0742 m) was obtained at very low heat fluxes (0.22 to 1.19 kw/so m). The system pressure and the bulk liquid subcooling varied in the range of 40 to 60 kPa and 3 to 5 C respectively. Thirty-eight boiling tests, each of 10 min duration for a given heat flux, were conducted. Measurements included the heater power, heater surface temperature, the liquid temperature and the system pressure as functions of heating time. Video data of the first 2 min of heating was recorded for each test. In some tests the video clearly shows the inception of boiling and the growth and departure of bubbles from the surface during the first 2 min of heating. In the absence of video data, the heater temperature variation during heating shows the inception of boiling and stable nucleate boiling. During the stable nucleate boiling, the wall superheat varied between 2.8 to 3.8 C for heat fluxes in the range of 0.95 to 1.19 kw/so m. The wall superheat at the inception of boiling varied between 2 to 13 C.

  20. Nucleate pool boiling in the long duration low gravity environment of the Space Shuttle

    NASA Technical Reports Server (NTRS)

    Hasan, M. M.; Lin, C. S.; Knoll, R. H.; Bentz, M. D.; Meserole, J. S.

    1993-01-01

    The results are presented of an experimental study of nucleate pool boiling performed in the low gravity environment of the space shuttle. Photographic observations of pool boiling in Freon 113 were obtained during the 'Tank Pressure Control Experiment,' flown on the Space Transportation System, STS-43 in August 1991. Nucleate boiling data from large (relative to bubble size) flat heating surfaces (0.1046 by 0.0742 m) was obtained at very low heat fluxes (0.22 to 1.19 kW/sq m). The system pressure and the bulk liquid subcooling varied in the range of 40 to 60 kPa and 3 to 5 C respectively. Thirty-eight boiling tests, each of 10-min duration for a given heat flux, were conducted. Measurements included the heater power, heater surface temperature, the liquid temperature and the system pressure as functions of heating time. Video data of the first 2 min of heating was recorded for each test. In some tests the video clearly shows the inception of boiling and the growth and departure of bubbles from the surface during the first 2 min of heating. In the absence of video data, the heater temperature variation during heating shows the inception of boiling and stable nucleate boiling. During the stable nucleate boiling, the wall superheat varied between 2.8 to 3.8 C for heat fluxes in the range of 0.95 to 1.19 kW/sq m. The wall superheat at the inception of boiling varied between 2 to 13 C.

  1. Structural studies and molecular dynamics simulations suggest a processive mechanism of exolytic lytic transglycosylase from Campylobacter jejuni.

    PubMed

    Vijayaraghavan, Jagamya; Kumar, Vijay; Krishnan, Nikhil P; Kaufhold, Ross T; Zeng, Ximin; Lin, Jun; van den Akker, Focco

    2018-01-01

    The bacterial soluble lytic transglycosylase (LT) breaks down the peptidoglycan (PG) layer during processes such as cell division. We present here crystal structures of the soluble LT Cj0843 from Campylobacter jejuni with and without bulgecin A inhibitor in the active site. Cj0843 has a doughnut shape similar but not identical to that of E. coli SLT70. The C-terminal catalytic domain is preceded by an L-domain, a large helical U-domain, a flexible linker, and a small N-terminal NU-domain. The flexible linker allows the NU-domain to reach over and complete the circular shape, using residues conserved in the Epsilonproteobacteria LT family. The inner surface of the Cj0843 doughnut is mostly positively charged including a pocket that has 8 Arg/Lys residues. Molecular dynamics simulations with PG strands revealed a potential functional role for this pocket in anchoring the negatively charged terminal tetrapeptide of the PG during several steps in the reaction including homing and aligning the PG strand for exolytic cleavage, and subsequent ratcheting of the PG strand to enhance processivity in degrading PG strands.

  2. Nucleate boiling performance on nano/microstructures with different wetting surfaces

    PubMed Central

    2012-01-01

    A study of nucleate boiling phenomena on nano/microstructures is a very basic and useful study with a view to the potential application of modified surfaces as heating surfaces in a number of fields. We present a detailed study of boiling experiments on fabricated nano/microstructured surfaces used as heating surfaces under atmospheric conditions, employing identical nanostructures with two different wettabilities (silicon-oxidized and Teflon-coated). Consequently, enhancements of both boiling heat transfer (BHT) and critical heat flux (CHF) are demonstrated in the nano/microstructures, independent of their wettability. However, the increment of BHT and CHF on each of the different wetting surfaces depended on the wetting characteristics of heating surfaces. The effect of water penetration in the surface structures by capillary phenomena is suggested as a plausible mechanism for the enhanced CHF on the nano/microstructures regardless of the wettability of the surfaces in atmospheric condition. This is supported by comparing bubble shapes generated in actual boiling experiments and dynamic contact angles under atmospheric conditions on Teflon-coated nano/microstructured surfaces. PMID:22559173

  3. 3. RW Meyer Sugar Mill: 18761889. Sorghum pan and boiling ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    3. RW Meyer Sugar Mill: 1876-1889. Sorghum pan and boiling range flue. Manufactured by John Nott & Co., Honolulu, Hawaii, 1878. View: South side of sorghum pan and boiling range flue. In the sorghum pan heat was applied to the cane juice to clarify it, evaporate its water content, and concentrate the sugar crystals. Hot gasses moved through the flue underneath the entire copper bottom of the sorghum pan from the furnace (east) end to the smokestack (west) end of the boiling range. The sorghum pan sides are of redwood. The flue is built of fire-brick, masonry, and portland cement. - R. W. Meyer Sugar Mill, State Route 47, Kualapuu, Maui County, HI

  4. Phase relations and adiabats in boiling seafloor geothermal systems

    USGS Publications Warehouse

    Bischoff, J.L.; Pitzer, Kenneth S.

    1985-01-01

    Observations of large salinity variations and vent temperatures in the range of 380-400??C suggest that boiling or two-phase separation may be occurring in some seafloor geothermal systems. Consideration of flow rates and the relatively small differences in density between vapors and liquids at the supercritical pressures at depth in these systems suggests that boiling is occurring under closed-system conditions. Salinity and temperature of boiling vents can be used to estimate the pressure-temperature point in the subsurface at which liquid seawater first reached the two-phase boundary. Data are reviewed to construct phase diagrams of coexisting brines and vapors in the two-phase region at pressures corresponding to those of the seafloor geothermal systems. A method is developed for calculating the enthalpy and entropy of the coexisting mixtures, and results are used to construct adiabats from the seafloor to the P-T two-phase boundary. Results for seafloor vents discharging at 2300 m below sea level indicate that a 385??C vent is composed of a brine (7% NaCl equivalent) in equilibrium with a vapor (0.1% NaCl). Brine constitutes 45% by weight of the mixture, and the fluid first boiled at approximately 1 km below the seafloor at 415??C, 330 bar. A 400??C vent is primarily vapor (88 wt.%, 0.044% NaCl) with a small amount of brine (26% NaCl) and first boiled at 2.9 km below the seafloor at 500??C, 520 bar. These results show that adiabatic decompression in the two-phase region results in dramatic cooling of the fluid mixture when there is a large fraction of vapor. ?? 1985.

  5. The sudden coalescene model of the boiling crisis

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Carrica, P.M.; Clausse, A.

    1995-09-01

    A local two-phase flow integral model of nucleate boiling and crisis is presented. The model is based on average balances on a control volume, yielding to a set of three nonlinear differential equations for the local void fraction, bubble number density and velocity. Boiling crisis as critical heat flux is interpreted as a dynamic transition caused by the coalescence of bubbles near the heater. The theoretical dynamic model is compared with experimental results obtained for linear power ramps in a horizontal plate heater in R-113, showing an excellent qualitative agreement.

  6. On Boiling of Crude Oil under Elevated Pressure

    NASA Astrophysics Data System (ADS)

    Pimenova, Anastasiya V.; Goldobin, Denis S.

    2016-02-01

    We construct a thermodynamic model for theoretical calculation of the boiling process of multicomponent mixtures of hydrocarbons (e.g., crude oil). The model governs kinetics of the mixture composition in the course of the distillation process along with the boiling temperature increase. The model heavily relies on the theory of dilute solutions of gases in liquids. Importantly, our results are applicable for modelling the process under elevated pressure (while the empiric models for oil cracking are not scalable to the case of extreme pressure), such as in an oil field heated by lava intrusions.

  7. Boiling points of halogenated ethanes: an explanatory model implicating weak intermolecular hydrogen-halogen bonding.

    PubMed

    Beauchamp, Guy

    2008-10-23

    This study explores via structural clues the influence of weak intermolecular hydrogen-halogen bonds on the boiling point of halogenated ethanes. The plot of boiling points of 86 halogenated ethanes versus the molar refraction (linked to polarizability) reveals a series of straight lines, each corresponding to one of nine possible arrangements of hydrogen and halogen atoms on the two-carbon skeleton. A multiple linear regression model of the boiling points could be designed based on molar refraction and subgroup structure as independent variables (R(2) = 0.995, standard error of boiling point 4.2 degrees C). The model is discussed in view of the fact that molar refraction can account for approximately 83.0% of the observed variation in boiling point, while 16.5% could be ascribed to weak C-X...H-C intermolecular interactions. The difference in the observed boiling point of molecules having similar molar refraction values but differing in hydrogen-halogen intermolecular bonds can reach as much as 90 degrees C.

  8. Nucleate pool boiling: High gravity to reduced gravity; liquid metals to cryogens

    NASA Technical Reports Server (NTRS)

    Merte, Herman, Jr.

    1988-01-01

    Requirements for the proper functioning of equipment and personnel in reduced gravity associated with space platforms and future space station modules introduce unique problems in temperature control; power generation; energy dissipation; the storage, transfer, control and conditioning of fluids; and liquid-vapor separation. The phase change of boiling is significant in all of these. Although both pool and flow boiling would be involved, research results to date include only pool boiling because buoyancy effects are maximized for this case. The effective application of forced convection boiling heat transfer in the microgravity of space will require a well grounded and cogent understanding of the mechanisms involved. Experimental results are presented for pool boiling from a single geometrical configuration, a flat surface, covering a wide range of body forces from a/g = 20 to 1 to a/g = 0 to -1 for a cryogenic liquid, and from a/g = 20 to 1 for water and a liquid metal. Similarities in behavior are noted for these three fluids at the higher gravity levels, and may reasonably be expected to continue at reduced gravity levels.

  9. Modeling acid-gas generation from boiling chloride brines

    PubMed Central

    2009-01-01

    Background This study investigates the generation of HCl and other acid gases from boiling calcium chloride dominated waters at atmospheric pressure, primarily using numerical modeling. The main focus of this investigation relates to the long-term geologic disposal of nuclear waste at Yucca Mountain, Nevada, where pore waters around waste-emplacement tunnels are expected to undergo boiling and evaporative concentration as a result of the heat released by spent nuclear fuel. Processes that are modeled include boiling of highly concentrated solutions, gas transport, and gas condensation accompanied by the dissociation of acid gases, causing low-pH condensate. Results Simple calculations are first carried out to evaluate condensate pH as a function of HCl gas fugacity and condensed water fraction for a vapor equilibrated with saturated calcium chloride brine at 50-150°C and 1 bar. The distillation of a calcium-chloride-dominated brine is then simulated with a reactive transport model using a brine composition representative of partially evaporated calcium-rich pore waters at Yucca Mountain. Results show a significant increase in boiling temperature from evaporative concentration, as well as low pH in condensates, particularly for dynamic systems where partial condensation takes place, which result in enrichment of HCl in condensates. These results are in qualitative agreement with experimental data from other studies. Conclusion The combination of reactive transport with multicomponent brine chemistry to study evaporation, boiling, and the potential for acid gas generation at the proposed Yucca Mountain repository is seen as an improvement relative to previously applied simpler batch evaporation models. This approach allows the evaluation of thermal, hydrological, and chemical (THC) processes in a coupled manner, and modeling of settings much more relevant to actual field conditions than the distillation experiment considered. The actual and modeled distillation

  10. Stability and potency of raw and boiled shrimp extracts for skin prick test.

    PubMed

    Pariyaprasert, Wipada; Piboonpocanun, Surapon; Jirapongsananuruk, Orathai; Visitsunthorn, Nualanong

    2015-06-01

    The difference of stability between raw and boiled shrimp extracts used in prick tests has never been investigated despite its potential consequences in tests development. The aim of this study was to compare the raw and boiled shrimp extracts of two species; Macrobrachium rosenbergii (freshwater shrimp) and Penaeus monodon (seawater shrimp) held at 4 ?C for different periods of time for their stability and potency in vivo by using the skin prick test (SPT) method. Raw and boiled M. rosenbergii and P. monodon extracts were prepared and stored at 4 ?C for 1, 7, 14 and 30 days. Thirty patients were pricked with raw and boiled shrimp extracts at all storage times, as well as prick to prick skin test (PTP) to fresh raw and boiled shrimps of both species. The mean wheal diameter (MWD) resulting from prick tests for all shrimp extracts was measured and compared. The shrimp extracts of all storage times yielded positive skin test results in the range of 90% - 100%. Raw P. monodon extracts induced larger wheals than boiled extracts at all storage times. There was no significant difference of MWD between raw and boiled M. rosenbergii extracts on day 1, 7, and 14. Significant correlations between MWD of PTP to fresh shrimps and SPT to all shrimp extracts were observed. All shrimp extracts were sterile at all storage times. Raw and boiled M. rosenbergii and P. monodon extracts were stable and sterile at 4 ?C for at most 30 days. SPT with these extracts induced more than 10 mm in shrimp allergy patients and the results were comparable with PTP to fresh shrimps.

  11. Identification of quantitative trait loci associated with boiled seed hardness in soybean

    PubMed Central

    Hirata, Kaori; Masuda, Ryoichi; Tsubokura, Yasutaka; Yasui, Takeshi; Yamada, Tetsuya; Takahashi, Koji; Nagaya, Taiko; Sayama, Takashi; Ishimoto, Masao; Hajika, Makita

    2014-01-01

    Boiled seed hardness is an important factor in the processing of soybean food products such as nimame and natto. Little information is available on the genetic basis for boiled seed hardness, despite the wide variation in this trait. DNA markers linked to the gene controlling this trait should be useful in soybean breeding programs because of the difficulty of its evaluation. In this report, quantitative trait locus (QTL) analysis was performed to reveal the genetic factors associated with boiled seed hardness using a recombinant inbred line population developed from a cross between two Japanese cultivars, ‘Natto-shoryu’ and ‘Hyoukei-kuro 3’, which differ largely in boiled seed hardness, which in ‘Natto-shoryu’ is about twice that of ‘Hyoukei-kuro 3’. Two significantly stable QTLs, qHbs3-1 and qHbs6-1, were identified on chromosomes 3 and 6, for which the ‘Hyoukei-kuro 3’ alleles contribute to decrease boiled seed hardness for both QTLs. qHbs3-1 also showed significant effects in progeny of a residual heterozygous line and in a different segregating population. Given its substantial effect on boiled seed hardness, SSR markers closely linked to qHbs3-1, such as BARCSOYSSR_03_0165 and BARCSOYSSR_03_0185, could be useful for marker-assisted selection in soybean breeding. PMID:25914591

  12. Flow Boiling and Condensation Experiment (FBCE) for the International Space Station

    NASA Technical Reports Server (NTRS)

    Mudawar, Issam; O'Neill, Lucas; Hasan, Mohammad; Nahra, Henry; Hall, Nancy; Balasubramaniam, R.; Mackey, Jeffrey

    2016-01-01

    An effective means to reducing the size and weight of future space vehicles is to replace present mostly single-phase thermal management systems with two-phase counterparts. By capitalizing upon both latent and sensible heat of the coolant rather than sensible heat alone, two-phase thermal management systems can yield orders of magnitude enhancement in flow boiling and condensation heat transfer coefficients. Because the understanding of the influence of microgravity on two-phase flow and heat transfer is quite limited, there is an urgent need for a new experimental microgravity facility to enable investigators to perform long-duration flow boiling and condensation experiments in pursuit of reliable databases, correlations and models. This presentation will discuss recent progress in the development of the Flow Boiling and Condensation Experiment (FBCE) for the International Space Station (ISS) in collaboration between Purdue University and NASA Glenn Research Center. Emphasis will be placed on the design of the flow boiling module and on new flow boiling data that were measured in parabolic flight, along with extensive flow visualization of interfacial features at heat fluxes up to critical heat flux (CHF). Also discussed a theoretical model that will be shown to predict CHF with high accuracy.

  13. Numerical Modeling of Propellant Boil-Off in a Cryogenic Storage Tank

    NASA Technical Reports Server (NTRS)

    Majumdar, A. K.; Steadman, T. E.; Maroney, J. L.; Sass, J. P.; Fesmire, J. E.

    2007-01-01

    A numerical model to predict boil-off of stored propellant in large spherical cryogenic tanks has been developed. Accurate prediction of tank boil-off rates for different thermal insulation systems was the goal of this collaboration effort. The Generalized Fluid System Simulation Program, integrating flow analysis and conjugate heat transfer for solving complex fluid system problems, was used to create the model. Calculation of tank boil-off rate requires simultaneous simulation of heat transfer processes among liquid propellant, vapor ullage space, and tank structure. The reference tank for the boil-off model was the 850,000 gallon liquid hydrogen tank at Launch Complex 39B (LC- 39B) at Kennedy Space Center, which is under study for future infrastructure improvements to support the Constellation program. The methodology employed in the numerical model was validated using a sub-scale model and tank. Experimental test data from a 1/15th scale version of the LC-39B tank using both liquid hydrogen and liquid nitrogen were used to anchor the analytical predictions of the sub-scale model. Favorable correlations between sub-scale model and experimental test data have provided confidence in full-scale tank boil-off predictions. These methods are now being used in the preliminary design for other cases including future launch vehicles

  14. Bubble dynamics, two-phase flow, and boiling heat transfer in a microgravity environment

    NASA Technical Reports Server (NTRS)

    Chung, Jacob N.

    1994-01-01

    The two-phase bubbly flow and boiling heat transfer in microgravity represents a substantial challenge to scientists and engineers and yet there is an urgent need to seek fundamental understanding in this area for future spacecraft design and space missions. At Washington State University, we have successfully designed, built and tested a 2.1 second drop tower with an innovation airbag deceleration system. Microgravity boiling experiments performed in our 0.6 second Drop Tower produced data flow visualizations that agree with published results and also provide some new understanding concerning flow boiling and microgravity bubble behavior. On the analytical and numerical work, the edge effects of finite divergent electrode plates on the forces experienced by bubbles were investigated. Boiling in a concentric cylinder microgravity and an electric field was numerically predicted. We also completed a feasibility study for microgravity boiling in an acoustic field.

  15. Boiling behavior of sodium-potassium alloy in a bench-scale solar receiver

    NASA Astrophysics Data System (ADS)

    Moreno, J. B.; Andraka, C. E.; Moss, T. A.

    During 1989-90, a 75-kW(sub t) sodium reflux pool-boiler solar receiver was successfully demonstrated at Sandia National Laboratories. Significant features of this receiver include the following: (1) boiling sodium as the heat transfer medium, and (2) electric-discharge-machined (EDM) cavities as artificial nucleation sites to stabilize boiling. Since this first demonstration, design of a second-generation pool-boiler receiver that will bring the concept closer to commercialization has begun. For long life, the new receiver uses Haynes Alloy 230. For increased safety factors against film boiling and flooding, it has a refined shape and somewhat larger dimensions. To eliminate the need for trace heating, the receiver will boil the sodium-potassium alloy NaK-78 instead of sodium. To reduce manufacturing costs, it will use one of a number of alternatives to EDM cavities for stabilization of boiling. To control incipient-boiling superheats, especially during hot restarts, it will contain a small amount of inert gas. Before the new receiver design could be finalized, bench-scale tests of some of the proposed changes were necessary. A series of bench-scale pool boilers were built from Haynes Alloy 230 and filled with NaK-78. Various boiling-stabilizer candidates were incorporated into them, including laser-drilled cavities and a number of different sintered-powder-metal coatings. These bench-scale pool boilers have been operated at temperatures up to 750 C, heated by quartz lamps with incident radiant fluxes up to 95 W/sq cm. The effects of various orientations and added gases have been studied. Results of these studies are presented.

  16. Characteristics of Pool Boiling on Graphite-Copper Composite Surfaces

    NASA Technical Reports Server (NTRS)

    Zhang, Nengli; Chao, David F.; Yang, Wen-Jei

    2002-01-01

    Nucleate pool boiling performance of different liquids on graphite-copper composite (Gr-Cu) surfaces has been experimentally studied and modeled. Both highly wetting fluids, such as freon-113 and pentane, and a moderately wetting fluid (water) were tested on the Gr-Cu surfaces with different graphite-fiber volume fractions to reveal the enhancement effects of the composite surfaces on the nucleate pool boiling. Results of the experiments show that the graphite-fiber volume fraction has an optimum value. The Gr-Cu composite surface with 25 percent graphite-fiber volume (f=0.25) has a maximum enhancement effect on the nucleate boiling heat transfer comparing to the pure copper surface. For the highly wetting fluid, the nucleate boiling heat transfer is generally enhanced on the Gr- Cu composite surfaces by 3 to 6 times shown. In the low heat flux region, the enhancement is over 6 times, but in the high heat flux region, the enhancement is reduced to about 40%. For the moderately wetting fluid (water), stronger enhancement of nucleate boiling heat transfer is achieved on the composite surface. It shown the experimental results in which one observes the nucleate boiling heat transfer enhancement of 5 to 10 times in the low heat flux region and an enhancement of 3 to 5 times in the high heat flux region. Photographs of bubble departure during the initial stage of nucleate boiling indicate that the bubbles detached from the composite surface are much smaller in diameter than those detached from the pure copper surface. Typical photographs are presented.It shows that the bubbles departed from the composite surface have diameters of only O(0.1) mm, while those departed from the pure copper surface have diameters of O(1) mm. It is also found that the bubbles depart from the composite surface at a much higher frequency, thus forming vapor columns. These two phenomena combined with high thermal conductivity of the graphite fiber are considered the mechanisms for such a

  17. Evaluation of Eukaryotic Cell Invasion on a Library of Genetically Diverse Campylobacter spp. Isolates.

    USDA-ARS?s Scientific Manuscript database

    Campylobacter spp. are the largest cause of sporadic bacterial gastrointestinal infection in the industrialized world. Epithelial cell invasion is thought to be necessary to bring about infection in humans. Invasion studies have shown that different Campylobacter jejuni isolates may differ in thei...

  18. Critical Heat Flux in Pool Boiling on Metal-Graphite Composite Surfaces

    NASA Technical Reports Server (NTRS)

    Zhang, Nengli; Yang, Wen-Jei; Chao, David F.; Chao, David F. (Technical Monitor)

    2000-01-01

    A study is conducted on high heat-flux pool boiling of pentane on micro-configured composite surfaces. The boiling surfaces are copper-graphite (Cu-Gr) and aluminum-graphite (Al-Gr) composites with a fiber volume concentration of 50%. The micro-graphite fibers embedded in the matrix contribute to a substantial enhancement in boiling heat-transfer performance. Correlation equations are obtained for both the isolated and coalesced bubble regimes, utilizing a mathematical model based on a metal-graphite, two-tier configuration with the aid of experimental data. A new model to predict the critical heat flux (CHF) on the composites is proposed to explain the fundamental aspects of the boiling phenomena. Three different factors affecting the CHF are considered in the model. Two of them are expected to become the main agents driving vapor volume detachment under microgravity conditions, using the metal-graphite composite surfaces as the heating surface and using liquids with an unusual Marangoni effect as the working fluid.

  19. Incorporating Water Boiling in the Numerical Modelling of Thermal Remediation by Electrical Resistance Heating

    NASA Astrophysics Data System (ADS)

    Molnar, I. L.; Krol, M.; Mumford, K. G.

    2017-12-01

    Developing numerical models for subsurface thermal remediation techniques - such as Electrical Resistive Heating (ERH) - that include multiphase processes such as in-situ water boiling, gas production and recovery has remained a significant challenge. These subsurface gas generation and recovery processes are driven by physical phenomena such as discrete and unstable gas (bubble) flow as well as water-gas phase mass transfer rates during bubble flow. Traditional approaches to multiphase flow modeling soil remain unable to accurately describe these phenomena. However, it has been demonstrated that Macroscopic Invasion Percolation (MIP) can successfully simulate discrete and unstable gas transport1. This has lead to the development of a coupled Electro Thermal-MIP Model2 (ET-MIP) capable of simulating multiple key processes in the thermal remediation and gas recovery process including: electrical heating of soil and groundwater, water flow, geological heterogeneity, heating-induced buoyant flow, water boiling, gas bubble generation and mobilization, contaminant mass transport and removal, and additional mechanisms such as bubble collapse in cooler regions. This study presents the first rigorous validation of a coupled ET-MIP model against two-dimensional water boiling and water/NAPL co-boiling experiments3. Once validated, the model was used to explore the impact of water and co-boiling events and subsequent gas generation and mobilization on ERH's ability to 1) generate, expand and mobilize gas at boiling and NAPL co-boiling temperatures, 2) efficiently strip contaminants from soil during both boiling and co-boiling. In addition, a quantification of the energy losses arising from steam generation during subsurface water boiling was examined with respect to its impact on the efficacy of thermal remediation. While this study specifically targets ERH, the study's focus on examining the fundamental mechanisms driving thermal remediation (e.g., water boiling) renders

  20. Radon depletion in xenon boil-off gas

    NASA Astrophysics Data System (ADS)

    Bruenner, S.; Cichon, D.; Lindemann, S.; Undagoitia, T. Marrodán; Simgen, H.

    2017-03-01

    An important background in detectors using liquid xenon for rare event searches arises from the decays of radon and its daughters. We report for the first time a reduction of ^{222}Rn in the gas phase above a liquid xenon reservoir. We show a reduction factor of ≳ 4 for the ^{222}Rn concentration in boil-off xenon gas compared to the radon enriched liquid phase. A semiconductor-based α -detector and miniaturized proportional counters are used to detect the radon. As the radon depletion in the boil-off gas is understood as a single-stage distillation process, this result establishes the suitability of cryogenic distillation to separate radon from xenon down to the 10^{-15} mol/mol level.

  1. 18. RW Meyer Sugar Mill: 18761889. Boiling House Interior, 1878. ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    18. RW Meyer Sugar Mill: 1876-1889. Boiling House Interior, 1878. View: Detail of floor with molasses pits below floor level. The remaining floor boards indicate the structure of the floor covering the entire inside of the boiling house. In the left background the base of the centrifugals are in view. - R. W. Meyer Sugar Mill, State Route 47, Kualapuu, Maui County, HI

  2. Forced convection flow boiling and two-phase flow phenomena in a microchannel

    NASA Astrophysics Data System (ADS)

    Na, Yun Whan

    2008-07-01

    The present study was performed to numerically analyze the evaporation phenomena through the liquid-vapor interface and to investigate bubble dynamics and heat transfer behavior during forced convective flow boiling in a microchannel. Flow instabilities of two-phase flow boiling in a microchannel were studied as well. The main objective of this research is to investigate the fundamental mechanisms of two-phase flow boiling in a microchannel and provide predictive tools to design thermal management systems, for example, microchannel heat sinks. The numerical results obtained from this study were qualitatively and quantitatively compared with experimental results in the open literature. Physical and mathematical models, accounting for evaporating phenomena through the liquid-vapor interface in a microchannel at constant heat flux and constant wall temperature, have been developed, respectively. The heat transfer mechanism is affected by the dominant heat conduction through the thin liquid film and vaporization at the liquid-vapor interface. The thickness of the liquid film and the pressure of the liquid and vapor phases were simultaneously solved by the governing differential equations. The developed semi-analytical evaporation model that takes into account of the interfacial phenomena and surface tension effects was used to obtain solutions numerically using the fourth-order Runge-Kutta method. The effects of heat flux 19 and wall temperature on the liquid film were evaluated. The obtained pressure drops in a microchannel were qualitatively consistent with the experimental results of Qu and Mudawar (2004). Forced convective flow boiling in a single microchannel with different channel heights was studied through a numerical simulation to investigate bubble dynamics, flow patterns, and heat transfer. The momentum and energy equations were solved using the finite volume method while the liquid-vapor interface of a bubble is captured using the VOF (Volume of Fluid

  3. Bubble Dynamics, Two-Phase Flow, and Boiling Heat Transfer in Microgravity

    NASA Technical Reports Server (NTRS)

    Chung, Jacob N.

    1998-01-01

    This report contains two independent sections. Part one is titled "Terrestrial and Microgravity Pool Boiling Heat Transfer and Critical heat flux phenomenon in an acoustic standing wave." Terrestrial and microgravity pool boiling heat transfer experiments were performed in the presence of a standing acoustic wave from a platinum wire resistance heater using degassed FC-72 Fluorinert liquid. The sound wave was created by driving a half wavelength resonator at a frequency of 10.15 kHz. Microgravity conditions were created using the 2.1 second drop tower on the campus of Washington State University. Burnout of the heater wire, often encountered with heat flux controlled systems, was avoided by using a constant temperature controller to regulate the heater wire temperature. The amplitude of the acoustic standing wave was increased from 28 kPa to over 70 kPa and these pressure measurements were made using a hydrophone fabricated with a small piezoelectric ceramic. Cavitation incurred during experiments at higher acoustic amplitudes contributed to the vapor bubble dynamics and heat transfer. The heater wire was positioned at three different locations within the acoustic field: the acoustic node, antinode, and halfway between these locations. Complete boiling curves are presented to show how the applied acoustic field enhanced boiling heat transfer and increased critical heat flux in microgravity and terrestrial environments. Video images provide information on the interaction between the vapor bubbles and the acoustic field. Part two is titled, "Design and qualification of a microscale heater array for use in boiling heat transfer." This part is summarized herein. Boiling heat transfer is an efficient means of heat transfer because a large amount of heat can be removed from a surface using a relatively small temperature difference between the surface and the bulk liquid. However, the mechanisms that govern boiling heat transfer are not well understood. Measurements of

  4. Steady State Vapor Bubble in Pool Boiling

    PubMed Central

    Zou, An; Chanana, Ashish; Agrawal, Amit; Wayner, Peter C.; Maroo, Shalabh C.

    2016-01-01

    Boiling, a dynamic and multiscale process, has been studied for several decades; however, a comprehensive understanding of the process is still lacking. The bubble ebullition cycle, which occurs over millisecond time-span, makes it extremely challenging to study near-surface interfacial characteristics of a single bubble. Here, we create a steady-state vapor bubble that can remain stable for hours in a pool of sub-cooled water using a femtosecond laser source. The stability of the bubble allows us to measure the contact-angle and perform in-situ imaging of the contact-line region and the microlayer, on hydrophilic and hydrophobic surfaces and in both degassed and regular (with dissolved air) water. The early growth stage of vapor bubble in degassed water shows a completely wetted bubble base with the microlayer, and the bubble does not depart from the surface due to reduced liquid pressure in the microlayer. Using experimental data and numerical simulations, we obtain permissible range of maximum heat transfer coefficient possible in nucleate boiling and the width of the evaporating layer in the contact-line region. This technique of creating and measuring fundamental characteristics of a stable vapor bubble will facilitate rational design of nanostructures for boiling enhancement and advance thermal management in electronics. PMID:26837464

  5. Detection, Isolation, and Molecular Subtyping of Escherichia coli O157:H7 and Campylobacter jejuni Associated with a Large Waterborne Outbreak

    PubMed Central

    Bopp, Dianna J.; Sauders, Brian D.; Waring, Alfred L.; Ackelsberg, Joel; Dumas, Nellie; Braun-Howland, Ellen; Dziewulski, David; Wallace, Barbara J.; Kelly, Molly; Halse, Tanya; Musser, Kimberlee Aruda; Smith, Perry F.; Morse, Dale L.; Limberger, Ronald J.

    2003-01-01

    The largest reported outbreak of waterborne Escherichia coli O157:H7 in the United States occurred in upstate New York following a county fair in August 1999. Culture methods were used to isolate E. coli O157:H7 from specimens from 128 of 775 patients with suspected infections. Campylobacter jejuni was also isolated from stools of 44 persons who developed diarrheal illness after attending this fair. There was one case of a confirmed coinfection with E. coli O157:H7 and C. jejuni. Molecular detection of stx1 and stx2 Shiga toxin genes, immunomagnetic separation (IMS), and selective culture enrichment were utilized to detect and isolate E. coli O157:H7 from an unchlorinated well and its distribution points, a dry well, and a nearby septic tank. PCR for stx1 and stx2 was shown to provide a useful screen for toxin-producing E. coli O157:H7, and IMS subculture improved recovery. Pulsed-field gel electrophoresis (PFGE) was used to compare patient and environmental E. coli O157:H7 isolates. Among patient isolates, 117 of 128 (91.5%) were type 1 or 1a (three or fewer bands different). Among the water distribution system isolates, 13 of 19 (68%) were type 1 or 1a. Additionally, PFGE of C. jejuni isolates revealed that 29 of 35 (83%) had indistinguishable PFGE patterns. The PFGE results implicated the water distribution system as the main source of the E. coli O157:H7 outbreak. This investigation demonstrates the potential for outbreaks involving more than one pathogen and the importance of analyzing isolates from multiple patients and environmental samples to develop a better understanding of bacterial transmission during an outbreak. PMID:12517844

  6. HISTOLOGICAL AND BIOCHEMICAL ANALYSIS OF MECHANICAL AND THERMAL BIOEFFECTS IN BOILING HISTOTRIPSY LESIONS INDUCED BY HIGH INTENSITY FOCUSED ULTRASOUND

    PubMed Central

    Wang, Yak-Nam; Khokhlova, Tatiana; Bailey, Michael; Hwang, Joo Ha; Khokhlova, Vera

    2013-01-01

    Recent studies have shown that shock wave heating and millisecond boiling in high intensity focused ultrasound (HIFU) fields can result in mechanical fractionation or emulsification of tissue - named boiling histotripsy. Visual observations of the change in color and contents indicated that the degree of thermal damage in the emulsified lesions can be controlled by varying the parameters of the exposure. The goal of this work was to examine thermal and mechanical effects in boiling histotripsy lesions using histological and biochemical analysis. The lesions were induced in ex vivo bovine heart and liver using a 2-MHz single-element transducer operating at duty factors of 0.005–0.01, pulse durations of 5–500 ms, and in situ shock amplitude of 73 MPa. Mechanical and thermal damage to tissue was evaluated histologically using conventional staining techniques (H&E and NADH-diphorase). Thermal effects were quantified by measuring denaturation of salt soluble proteins in the treated region. According to histology, the lesions that visually appeared as a liquid, contained no cellular structures larger than a cell nucleus and had a very sharp border of 1–2 cells. Both histology and protein analysis showed that lesions obtained with short pulses (< 10 ms) did not contain any thermal damage. Increasing the pulse duration resulted in an increase in thermal damage. However, both protein analysis and NADH-diaphorase staining showed less denaturation than visually observed as whitening of tissue. The number of HIFU pulses delivered per exposure did not change the lesion shape or the degree of thermal denaturation, whereas the size of the lesion showed a saturating behaviour thus suggesting optimal exposure duration. This study confirmed that boiling histotripsy offers an effective, predictable way to non-invasively fractionate tissue into subcellular fragments with or without inducing thermal damage. PMID:23312958

  7. Estimating surface temperature in forced convection nucleate boiling - A simplified method

    NASA Technical Reports Server (NTRS)

    Hendricks, R. C.; Papell, S. S.

    1977-01-01

    A simplified expression to estimate surface temperatures in forced convection boiling was developed using a liquid nitrogen data base. Using the principal of corresponding states and the Kutateladze relation for maximum pool boiling heat flux, the expression was normalized for use with other fluids. The expression was applied also to neon and water. For the neon data base, the agreement was acceptable with the exclusion of one set suspected to be in the transition boiling regime. For the water data base at reduced pressure greater than 0.05 the agreement is generally good. At lower reduced pressures, the water data scatter and the calculated temperature becomes a function of flow rate.

  8. Antimicrobial, Rheological, and Thermal Properties of Plasticized Polylactide Films Incorporated with Essential Oils to Inhibit Staphylococcus aureus and Campylobacter jejuni.

    PubMed

    Ahmed, Jasim; Hiremath, Nikhil; Jacob, Harsha

    2016-02-01

    Polylactide (PLA) is the most mature biobased and biodegradable polymer. Due to its inherent brittleness, the polymer cannot be used as a packaging material without plasticizer. An attempt was made to develop antimicrobial plasticized PLA film by incorporating polyethylene glycol (PEG) and 3 essential oils (EO), namely cinnamon, garlic, and clove by solvent casting method. Physical, thermal, and rheological properties of those films were evaluated for practical applications whereas the antimicrobial properties were tested against Staphylococcus aureus and Campylobacter jejuni-pathogens related to poultry industry. Both PEG and EOs led to the formation of flexible PLA/PEG/EO films with significant drop in the glass transition temperature (Tg ), and mechanical property. Time-temperature superposition (TTS) principle was employed to melt rheology of EO-based films at selected temperature, and rheological moduli superimposed well in an extended frequency range. Among EOs, cinnamon and clove oil-based films (PLA/PEG/CIN and PLA/PEG/CLO) exhibited a complete zone of inhibition against C. jejuni at the maximum concentration (1.6 mL per 2 g PLA/PEG blend) whereas the garlic oil-based film (PLA/PEG/GAR) had the lowest activity. © 2016 Institute of Food Technologists®

  9. Recent Work on Flow Boiling and Condensation in a Single Microchannel

    NASA Astrophysics Data System (ADS)

    Quan, Xiaojun; Wang, Guodong; Cheng, Ping; Wu, Huiying

    2007-06-01

    Recent visualization and measurements results on flow boiling of water and condensation of steam in a single microchannel, carried out at Shanghai Jiaotong University, is summarized in this paper. For flow boiling of water, experiments were conducted in a single microchannel with a trapezoidal cross-section having a hydraulic diameter of 186 μm and a length of 30 mm. A boiling flow pattern map in terms of heat flux versus mass flux, showing the unstable and stable boiling flow regimes in the microchannel, is obtained. For the investigation of condensation, experiments were carried out for steam condensing inside a single microchannel with a length of 60mm having a hydraulic diameter of 87 μm and 120μm respectively. The location of transition from annular flow to plug/slug flow in a microchannel is found to be dependent on both the dimensionless condensation heat transfer rate as well as the Reynolds number of the steam. The frequency for the occurrence of the injection flow is found to increase with the increasing mass flux.

  10. Critical heat flux for free convection boiling in thin rectangular channels

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Cheng, Lap Y.; Tichler, P.R.

    A review of the experimental data on free convection boiling critical heat flux (CHF) in vertical rectangular channels reveals three mechanisms of burnout. They are the pool boiling limit, the circulation limit, and the flooding limit associated with a transition in flow regime from churn to annular flow. The dominance of a particular mechanism depends on the dimensions of the channel. Analytical models were developed for each free convection boiling limit. Limited agreement with data is observed. A CHF correlation, which is valid for a wide range of gap sizes, was constructed from the CHFs calculated according to the threemore » mechanisms of burnout. 17 refs., 7 figs.« less

  11. Sensory quality and appropriateness of raw and boiled Jerusalem artichoke tubers (Helianthus tuberosus L.).

    PubMed

    Bach, Vibe; Kidmose, Ulla; Thybo, Anette K; Edelenbos, Merete

    2013-03-30

    The aim of the present study was to investigate the sensory attributes, dry matter and sugar content of five varieties of Jerusalem artichoke tubers and their relation to the appropriateness of the tubers for raw and boiled preparation. Sensory evaluation of raw and boiled Jerusalem artichoke tubers was performed by a trained sensory panel and a semi-trained consumer panel of 49 participants, who also evaluated the appropriateness of the tubers for raw and boiled preparation. The appropriateness of raw Jerusalem artichoke tubers was related to Jerusalem artichoke flavour, green nut flavour, sweetness and colour intensity, whereas the appropriateness of boiled tubers was related to celeriac aroma, sweet aroma, sweetness and colour intensity. In both preparations the variety Dwarf stood out from the others by being the least appropriate tuber. A few sensory attributes can be used as predictors of the appropriateness of Jerusalem artichoke tubers for raw and boiled consumption. Knowledge on the quality of raw and boiled Jerusalem artichoke tubers can be used to inform consumers on the right choice of raw material and thereby increase the consumption of the vegetable. © 2012 Society of Chemical Industry.

  12. Increasing Boiling Heat Transfer using Low Conductivity Materials

    PubMed Central

    Mahamudur Rahman, Md; Pollack, Jordan; McCarthy, Matthew

    2015-01-01

    We report the counterintuitive mechanism of increasing boiling heat transfer by incorporating low-conductivity materials at the interface between the surface and fluid. By embedding an array of non-conductive lines into a high-conductivity substrate, in-plane variations in the local surface temperature are created. During boiling the surface temperature varies spatially across the substrate, alternating between high and low values, and promotes the organization of distinct liquid and vapor flows. By systematically tuning the peak-to-peak wavelength of this spatial temperature variation, a resonance-like effect is seen at a value equal to the capillary length of the fluid. Replacing ~18% of the surface with a non-conductive epoxy results in a greater than 5x increase in heat transfer rate at a given superheat temperature. This drastic and counterintuitive increase is shown to be due to optimized bubble dynamics, where ordered pathways allow for efficient removal of vapor and the return of replenishing liquid. The use of engineered thermal gradients represents a potentially disruptive approach to create high-efficiency and high-heat-flux boiling surfaces which are naturally insensitive to fouling and degradation as compared to other approaches. PMID:26281890

  13. Evaluation of correlations of flow boiling heat transfer of R22 in horizontal channels.

    PubMed

    Zhou, Zhanru; Fang, Xiande; Li, Dingkun

    2013-01-01

    The calculation of two-phase flow boiling heat transfer of R22 in channels is required in a variety of applications, such as chemical process cooling systems, refrigeration, and air conditioning. A number of correlations for flow boiling heat transfer in channels have been proposed. This work evaluates the existing correlations for flow boiling heat transfer coefficient with 1669 experimental data points of flow boiling heat transfer of R22 collected from 18 published papers. The top two correlations for R22 are those of Liu and Winterton (1991) and Fang (2013), with the mean absolute deviation of 32.7% and 32.8%, respectively. More studies should be carried out to develop better ones. Effects of channel dimension and vapor quality on heat transfer are analyzed, and the results provide valuable information for further research in the correlation of two-phase flow boiling heat transfer of R22 in channels.

  14. Numerical Investigation of Microgravity Tank Pressure Rise Due to Boiling

    NASA Technical Reports Server (NTRS)

    Hylton, Sonya; Ibrahim, Mounir; Kartuzova, Olga; Kassemi, Mohammad

    2015-01-01

    The ability to control self-pressurization in cryogenic storage tanks is essential for NASAs long-term space exploration missions. Predictions of the tank pressure rise in Space are needed in order to inform the microgravity design and optimization process. Due to the fact that natural convection is very weak in microgravity, heat leaks into the tank can create superheated regions in the liquid. The superheated regions can instigate microgravity boiling, giving rise to pressure spikes during self-pressurization. In this work, a CFD model is developed to predict the magnitude and duration of the microgravity pressure spikes. The model uses the Schrage equation to calculate the mass transfer, with a different accommodation coefficient for evaporation at the interface, condensation at the interface, and boiling in the bulk liquid. The implicit VOF model was used to account for the moving interface, with bounded second order time discretization. Validation of the models predictions was carried out using microgravity data from the Tank Pressure Control Experiment, which flew aboard the Space Shuttle Mission STS-52. Although this experiment was meant to study pressurization and pressure control, it underwent boiling during several tests. The pressure rise predicted by the CFD model compared well with the experimental data. The ZBOT microgravity experiment is scheduled to fly on February 2016 aboard the ISS. The CFD model was also used to perform simulations for setting parametric limits for the Zero-Boil-Off Tank (ZBOT) Experiments Test Matrix in an attempt to avoid boiling in the majority of the test runs that are aimed to study pressure increase rates during self-pressurization. *Supported in part by NASA ISS Physical Sciences Research Program, NASA HQ, USA

  15. Flow-Boiling Critical Heat Flux Experiments Performed in Reduced Gravity

    NASA Technical Reports Server (NTRS)

    Hasan, Mohammad M.; Mudawar, Issam

    2005-01-01

    Poor understanding of flow boiling in microgravity has recently emerged as a key obstacle to the development of many types of power generation and advanced life support systems intended for space exploration. The critical heat flux (CHF) is perhaps the most important thermal design parameter for boiling systems involving both heatflux-controlled devices and intense heat removal. Exceeding the CHF limit can lead to permanent damage, including physical burnout of the heat-dissipating device. The importance of the CHF limit creates an urgent need to develop predictive design tools to ensure both the safe and reliable operation of a two-phase thermal management system under the reduced-gravity (like that on the Moon and Mars) and microgravity environments of space. At present, very limited information is available on flow-boiling heat transfer and the CHF under these conditions.

  16. Science 101: Why Does It Take Longer to Boil Potatoes at High Altitudes?

    ERIC Educational Resources Information Center

    Robertson, Bill

    2017-01-01

    Why Does It Take Longer to Boil Potatoes at High Altitudes? This column provides background science information for elementary teachers. This month's issue looks at why water boils at different temperatures at different altitudes.

  17. 16. RW Meyer Sugar Mill: 18761889. Boiling House Interior, 1878. ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    16. RW Meyer Sugar Mill: 1876-1889. Boiling House Interior, 1878. View: Looking from west to east through boiling house. The sorghum pan is on the right. The beams; joists, and trusses are of northwest pine; side boards are of redwood. A foundation line of a loading dock and smokestack are in the foreground. Both end walls have deteriorated completely. - R. W. Meyer Sugar Mill, State Route 47, Kualapuu, Maui County, HI

  18. Effect of soaking, boiling, and steaming on total phenolic contentand antioxidant activities of cool season food legumes.

    PubMed

    Xu, Baojun; Chang, Sam K C

    2008-09-01

    The effects of soaking, boiling and steaming processes on the total phenolic components and antioxidant activity in commonly consumed cool season food legumes (CSFL's), including green pea, yellow pea, chickpea and lentil were investigated. As compared to original unprocessed legumes, all processing steps caused significant (p<0.05) decreases in total phenolic content (TPC), DPPH free radical scavenging activity (DPPH) in all tested CSFL's. All soaking and atmospheric boiling treatments caused significant (p<0.05) decreases in oxygen radical absorbing capacity (ORAC). However, pressure boiling and pressure steaming caused significant (p<0.05) increases in ORAC values. Steaming treatments resulted in a greater retention of TPC, DPPH, and ORAC values in all tested CSFL's as compared to boiling treatments. To obtain cooked legumes with similar palatability and firmness, pressure boiling shortened processing time as compared to atmospheric boiling, resulted in insignificant differences in TPC, DPPH for green and yellow pea. However, TPC and DPPH in cooked lentils differed significantly between atmospheric and pressure boiling. As compared to atmospheric processes, pressure processes significantly increased ORAC values in both boiled and steamed CSFL's. Greater TPC, DPPH and ORAC values were detected in boiling water than that in soaking and steaming water. Boiling also caused more solid loss than steaming. Steam processing exhibited several advantages in retaining the integrity of the legume appearance and texture of the cooked product, shortening process time, and greater retention of antioxidant components and activities. Copyright © 2008 Elsevier Ltd. All rights reserved.

  19. Enhanced Droplet Control by Transition Boiling

    PubMed Central

    Grounds, Alex; Still, Richard; Takashina, Kei

    2012-01-01

    A droplet of water on a heated surface can levitate over a film of gas produced by its own evaporation in the Leidenfrost effect. When the surface is prepared with ratchet-like saw-teeth topography, these droplets can self-propel and can even climb uphill. However, the extent to which the droplets can be controlled is limited by the physics of the Leidenfrost effect. Here, we show that transition boiling can be induced even at very high surface temperatures and provide additional control over the droplets. Ratchets with acute protrusions enable droplets to climb steeper inclines while ratchets with sub-structures enable their direction of motion to be controlled by varying the temperature of the surface. The droplets' departure from the Leidenfrost regime is assessed by analysing the sound produced by their boiling. We anticipate these techniques will enable the development of more sophisticated methods for controlling small droplets and heat transfer. PMID:23056912

  20. Enhanced Droplet Control by Transition Boiling

    NASA Astrophysics Data System (ADS)

    Grounds, Alex; Still, Richard; Takashina, Kei

    2012-10-01

    A droplet of water on a heated surface can levitate over a film of gas produced by its own evaporation in the Leidenfrost effect. When the surface is prepared with ratchet-like saw-teeth topography, these droplets can self-propel and can even climb uphill. However, the extent to which the droplets can be controlled is limited by the physics of the Leidenfrost effect. Here, we show that transition boiling can be induced even at very high surface temperatures and provide additional control over the droplets. Ratchets with acute protrusions enable droplets to climb steeper inclines while ratchets with sub-structures enable their direction of motion to be controlled by varying the temperature of the surface. The droplets' departure from the Leidenfrost regime is assessed by analysing the sound produced by their boiling. We anticipate these techniques will enable the development of more sophisticated methods for controlling small droplets and heat transfer.

  1. Evaluation of Correlations of Flow Boiling Heat Transfer of R22 in Horizontal Channels

    PubMed Central

    Fang, Xiande; Li, Dingkun

    2013-01-01

    The calculation of two-phase flow boiling heat transfer of R22 in channels is required in a variety of applications, such as chemical process cooling systems, refrigeration, and air conditioning. A number of correlations for flow boiling heat transfer in channels have been proposed. This work evaluates the existing correlations for flow boiling heat transfer coefficient with 1669 experimental data points of flow boiling heat transfer of R22 collected from 18 published papers. The top two correlations for R22 are those of Liu and Winterton (1991) and Fang (2013), with the mean absolute deviation of 32.7% and 32.8%, respectively. More studies should be carried out to develop better ones. Effects of channel dimension and vapor quality on heat transfer are analyzed, and the results provide valuable information for further research in the correlation of two-phase flow boiling heat transfer of R22 in channels. PMID:23956695

  2. Bench-scale screening tests for a boiling sodium-potassium alloy solar receiver

    NASA Astrophysics Data System (ADS)

    Moreno, J. B.; Moss, T. A.

    1993-06-01

    Bench-scale tests were carried out in support of the design of a second-generation 75-kW(sub t) reflux pool-boiler solar receiver. The receiver will be made from Haynes Alloy 230 and will contain the sodium-potassium alloy NaK-78. The bench-scale tests used quartz lamp heated boilers to screen candidate boiling stabilization materials and methods at temperatures up to 750 degree C. Candidates that provided stable boiling were tested for hot-restart behavior. Poor stability was obtained with single 1/4-inch diameter patches of powdered metal hot press sintered onto the wetted side of the heat-input area. Laser-drilled and electric discharge machined cavities in the heated surface also performed poorly. Small additions of xenon, and heated-surface tilt out of the vertical, dramatically improved poor boiling stability; additions of helium or oxygen did not. The most stable boiling was obtained when the entire heat-input area was covered by a powdered-metal coating. The effect of heated-area size was assessed for one coating: at low incident fluxes, when even this coating performed poorly, increasing the heated-area size markedly improved boiling stability. Good hot-restart behavior was not observed with any candidate, although results were significantly better with added xenon in a boiler shortened from 3 to 2 feet. In addition to the screening tests, flash-radiography imaging of metal-vapor bubbles during boiling was attempted. Contrary to the Cole-Rohsenow correlation, these bubble-size estimates did not vary with pressure; instead they were constant, consistent with the only other alkali metal measurements, but about 1/2 their size.

  3. Controlled tissue emulsification produced by high intensity focused ultrasound shock waves and millisecond boiling

    PubMed Central

    Khokhlova, Tatiana D.; Canney, Michael S.; Khokhlova, Vera A.; Sapozhnikov, Oleg A.; Crum, Lawrence A.; Bailey, Michael R.

    2011-01-01

    In high intensity focused ultrasound (HIFU) applications, tissue may be thermally necrosed by heating, emulsified by cavitation, or, as was recently discovered, emulsified using repetitive millisecond boiling caused by shock wave heating. Here, this last approach was further investigated. Experiments were performed in transparent gels and ex vivo bovine heart tissue using 1, 2, and 3 MHz focused transducers and different pulsing schemes in which the pressure, duty factor, and pulse duration were varied. A previously developed derating procedure to determine in situ shock amplitudes and the time-to-boil was refined. Treatments were monitored using B-mode ultrasound. Both inertial cavitation and boiling were observed during exposures, but emulsification occurred only when shocks and boiling were present. Emulsified lesions without thermal denaturation were produced with shock amplitudes sufficient to induce boiling in less than 20 ms, duty factors of less than 0.02, and pulse lengths shorter than 30 ms. Higher duty factors or longer pulses produced varying degrees of thermal denaturation combined with mechanical emulsification. Larger lesions were obtained using lower ultrasound frequencies. The results show that shock wave heating and millisecond boiling is an effective and reliable way to emulsify tissue while monitoring the treatment with ultrasound. PMID:22088025

  4. Transient boiling in two-phase helium natural circulation loops

    NASA Astrophysics Data System (ADS)

    Furci, H.; Baudouy, B.; Four, A.; Meuris, C.

    2014-01-01

    Two-phase helium natural circulation loops are used for cooling large superconducting magnets, as CMS for LHC. During normal operation or in the case of incidents, transients are exerted on the cooling system. Here a cooling system of this type is studied experimentally. Sudden power changes are operated on a vertical-heated-section natural convection loop, simulating a fast increase of heat deposition on magnet cooling pipes. Mass flow rate, heated section wall temperature and pressure drop variations are measured as a function of time, to assess the time behavior concerning the boiling regime according to the values of power injected on the heated section. The boiling curves and critical heat flux (CHF) values have been obtained in steady state. Temperature evolution has been observed in order to explore the operating ranges where heat transfer is deteriorated. Premature film boiling has been observed during transients on the heated section in some power ranges, even at appreciably lower values than the CHF. A way of attenuating these undesired temperature excursions has been identified through the application of high enough initial heating power.

  5. Boiling of an emulsion in a yield stress fluid.

    PubMed

    Guéna, Geoffroy; Wang, Ji; d'Espinose, Jean-Baptiste; Lequeux, François; Talini, Laurence

    2010-11-01

    We report the boiling behavior of pentane emulsified in a yield stress fluid, a colloidal clay (Laponite) suspension. We have observed that a superheated state is easily reached: the emulsion, heated more than 50 °C above the alkane boiling point, does not boil. Superheating is made possible by the suppression of heterogeneous nucleation in pentane, resulting from the emulsification process, a phenomenon evidenced decades ago in studies of the superheating of two phase fluids. We have furthermore studied the growth of isolated bubbles nucleated in the emulsion. The rate of increase of the bubble radius with time depends on both the temperature and emulsion volume fraction but, rather unexpectedly, does not depend on the fluid rheology. We show that the bubbles grow by diffusion of the alkane through the aqueous phase between liquid droplets and bubbles, analogously to an Ostwald ripening process. The peculiarity of the process reported here is that a layer depleted in oil droplets forms around the bubble, layer to which the alkane concentration gradient is confined. We successfully describe our experimental results with a simple transfer model.

  6. Stability of film boiling on inclined plates and spheres

    NASA Astrophysics Data System (ADS)

    Aursand, Eskil; Hammer, Morten; Munkejord, Svend Tollak; Müller, Bernhard; Ytrehus, Tor

    2017-11-01

    In film boiling, a continuous sub-millimeter vapor film forms between a liquid and a heated surface, insulating the two from each other. While quite accurate steady state solutions are readily obtained, the intermediate Reynolds numbers can make transient analysis challenging. The present work is a theoretical study of film boiling instabilities. We study the formation of travelling waves that are a combination of Kelvin-Helmholtz and the Rayleigh-Taylor instabilities. In particular, we study how the nature of this process depends on the Reynolds number, the Bond number, and the inclination of the submerged heated plate. In addition we extend the analysis to the case of a submerged heated sphere. Modelling of the transient dynamics of such films is important for answering practical questions such as how instabilities affect the overall heat transfer, and whether they can lead to complete film boiling collapse (Leidenfrost point). This work has been financed under the MAROFF program. We acknowledge the Research Council of Norway (244076/O80) and The Gas Technology Centre NTNU-SINTEF (GTS) for support.

  7. Large-scale Generation of Patterned Bubble Arrays on Printed Bi-functional Boiling Surfaces

    PubMed Central

    Choi, Chang-Ho; David, Michele; Gao, Zhongwei; Chang, Alvin; Allen, Marshall; Wang, Hailei; Chang, Chih-hung

    2016-01-01

    Bubble nucleation control, growth and departure dynamics is important in understanding boiling phenomena and enhancing nucleate boiling heat transfer performance. We report a novel bi-functional heterogeneous surface structure that is capable of tuning bubble nucleation, growth and departure dynamics. For the fabrication of the surface, hydrophobic polymer dot arrays are first printed on a substrate, followed by hydrophilic ZnO nanostructure deposition via microreactor-assisted nanomaterial deposition (MAND) processing. Wettability contrast between the hydrophobic polymer dot arrays and aqueous ZnO solution allows for the fabrication of heterogeneous surfaces with distinct wettability regions. Heterogeneous surfaces with various configurations were fabricated and their bubble dynamics were examined at elevated heat flux, revealing various nucleate boiling phenomena. In particular, aligned and patterned bubbles with a tunable departure frequency and diameter were demonstrated in a boiling experiment for the first time. Taking advantage of our fabrication method, a 6 inch wafer size heterogeneous surface was prepared. Pool boiling experiments were also performed to demonstrate a heat flux enhancement up to 3X at the same surface superheat using bi-functional surfaces, compared to a bare stainless steel surface. PMID:27034255

  8. Large-scale Generation of Patterned Bubble Arrays on Printed Bi-functional Boiling Surfaces

    NASA Astrophysics Data System (ADS)

    Choi, Chang-Ho; David, Michele; Gao, Zhongwei; Chang, Alvin; Allen, Marshall; Wang, Hailei; Chang, Chih-Hung

    2016-04-01

    Bubble nucleation control, growth and departure dynamics is important in understanding boiling phenomena and enhancing nucleate boiling heat transfer performance. We report a novel bi-functional heterogeneous surface structure that is capable of tuning bubble nucleation, growth and departure dynamics. For the fabrication of the surface, hydrophobic polymer dot arrays are first printed on a substrate, followed by hydrophilic ZnO nanostructure deposition via microreactor-assisted nanomaterial deposition (MAND) processing. Wettability contrast between the hydrophobic polymer dot arrays and aqueous ZnO solution allows for the fabrication of heterogeneous surfaces with distinct wettability regions. Heterogeneous surfaces with various configurations were fabricated and their bubble dynamics were examined at elevated heat flux, revealing various nucleate boiling phenomena. In particular, aligned and patterned bubbles with a tunable departure frequency and diameter were demonstrated in a boiling experiment for the first time. Taking advantage of our fabrication method, a 6 inch wafer size heterogeneous surface was prepared. Pool boiling experiments were also performed to demonstrate a heat flux enhancement up to 3X at the same surface superheat using bi-functional surfaces, compared to a bare stainless steel surface.

  9. Transient boiling heat transfer in saturated liquid nitrogen and F113 at standard and zero gravity

    NASA Technical Reports Server (NTRS)

    Oker, E.; Merte, H., Jr.

    1973-01-01

    Transient and steady state nucleate boiling in saturated LN2 and F113 at standard and near zero gravity conditions were investigated for the horizontal up, vertical and horizontal down orientations of the heating surface. Two distinct regimes of heat transfer mechanisms were observed during the interval from the step increase of power input to the onset of nucleate boiling: the conduction and convection dominated regimes. The time duration in each regime was considerably shorter with LN2 than with F113, and decreased as heat flux increased, as gravity was reduced, and as the orientation was changed from horizontal up to horizontal down. In transient boiling, boiling initiates at a single point following the step increase in power, and then spreads over the surface. The delay time for the inception of boiling at the first site, and the velocity of spread of boiling varies depending upon the heat flux, orientation, body force, surface roughness and liquid properties, and are a consequence of changes in boundary layer temperature levels associated with changes in natural convection. Following the step increase in power input, surface temperature overshoot and undershoot occur before the steady state boiling temperature level is established.

  10. Assessing the microbiological performance and potential cost of boiling drinking water in urban Zambia.

    PubMed

    Psutka, Rebecca; Peletz, Rachel; Michelo, Sandford; Kelly, Paul; Clasen, Thomas

    2011-07-15

    Boiling is the most common method of disinfecting water in the home and the benchmark against which other point-of-use water treatment is measured. In a six-week study in peri-urban Zambia, we assessed the microbiological effectiveness and potential cost of boiling among 49 households without a water connection who reported "always" or "almost always" boiling their water before drinking it. Source and household drinking water samples were compared weekly for thermotolerant coliforms (TTC), an indicator of fecal contamination. Demographics, costs, and other information were collected through surveys and structured observations. Drinking water samples taken at the household (geometric mean 7.2 TTC/100 mL, 95% CI, 5.4-9.7) were actually worse in microbiological quality than source water (geometric mean 4.0 TTC/100 mL, 95% CI, 3.1-5.1) (p < 0.001), although both are relatively low levels of contamination. Only 60% of drinking water samples were reported to have actually been boiled at the time of collection from the home, suggesting over-reporting and inconsistent compliance. However, these samples were of no higher microbiological quality. Evidence suggests that water quality deteriorated after boiling due to lack of residual protection and unsafe storage and handling. The potential cost of fuel or electricity for boiling was estimated at 5% and 7% of income, respectively. In this setting where microbiological water quality was relatively good at the source, safe-storage practices that minimize recontamination may be more effective in managing the risk of disease from drinking water at a fraction of the cost of boiling.

  11. Use of the potential probiotic strain Lactobacillus salivarius SMXD51 to control Campylobacter jejuni in broilers.

    PubMed

    Saint-Cyr, Manuel Jimmy; Haddad, Nabila; Taminiau, Bernard; Poezevara, Typhaine; Quesne, Ségolène; Amelot, Michel; Daube, Georges; Chemaly, Marianne; Dousset, Xavier; Guyard-Nicodème, Muriel

    2017-04-17

    Campylobacteriosis is the most frequently reported zoonotic disease in humans in the EU since 2005. As chicken meat is the main source of contamination, reducing the level of Campylobacter in broiler chicken will lower the risk to consumers. The aim of this project was to evaluate the ability of Lactobacillus salivarius SMXD51 to control Campylobacter jejuni in broilers and to investigate the mechanisms that could be involved. Thirty broilers artificially contaminated with C. jejuni were treated by oral gavage with MRS broth or a bacterial suspension (10 7 CFU) of Lb. salivarius SMXD51 (SMXD51) in MRS broth. At 14 and 35days of age, Campylobacter and Lb. salivarius loads were assessed in cecal contents. The impact of the treatment on the avian gut microbiota at day 35 was also evaluated. At day 14, the comparison between the control and treated groups showed a significant reduction (P<0.05) of 0.82 log. After 35days, a significant reduction (P<0.001) of 2.81 log in Campylobacter loads was observed and 73% of chickens treated with the culture exhibited Campylobacter loads below 7log 10 CFU/g. Taxonomic analysis revealed that SMXD51 treatment induced significant changes (P<0.05) in a limited number of bacterial genera of the avian gut microbiota and partially limited the impact of Campylobacter on Anaerotruncus sp. decrease and Subdoligranulum sp. increase. Thus, SMXD51 exhibits an anti-Campylobacter activity in vivo and can partially prevent the impact of Campylobacter on the avian gut microbiota. Copyright © 2016 Elsevier B.V. All rights reserved.

  12. Multiphysics modeling of two-phase film boiling within porous corrosion deposits

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Jin, Miaomiao, E-mail: mmjin@mit.edu; Short, Michael, E-mail: hereiam@mit.edu

    2016-07-01

    Porous corrosion deposits on nuclear fuel cladding, known as CRUD, can cause multiple operational problems in light water reactors (LWRs). CRUD can cause accelerated corrosion of the fuel cladding, increase radiation fields and hence greater exposure risk to plant workers once activated, and induce a downward axial power shift causing an imbalance in core power distribution. In order to facilitate a better understanding of CRUD's effects, such as localized high cladding surface temperatures related to accelerated corrosion rates, we describe an improved, fully-coupled, multiphysics model to simulate heat transfer, chemical reactions and transport, and two-phase fluid flow within these deposits.more » Our new model features a reformed assumption of 2D, two-phase film boiling within the CRUD, correcting earlier models' assumptions of single-phase coolant flow with wick boiling under high heat fluxes. This model helps to better explain observed experimental values of the effective CRUD thermal conductivity. Finally, we propose a more complete set of boiling regimes, or a more detailed mechanism, to explain recent CRUD deposition experiments by suggesting the new concept of double dryout specifically in thick porous media with boiling chimneys. - Highlights: • A two-phase model of CRUD's effects on fuel cladding is developed and improved. • This model eliminates the formerly erroneous assumption of wick boiling. • Higher fuel cladding temperatures are predicted when accounting for two-phase flow. • Double-peaks in thermal conductivity vs. heat flux in experiments are explained. • A “double dryout” mechanism in CRUD is proposed based on the model and experiments.« less

  13. Single-bubble dynamics in pool boiling of one-component fluids.

    PubMed

    Xu, Xinpeng; Qian, Tiezheng

    2014-06-01

    We numerically investigate the pool boiling of one-component fluids with a focus on the effects of surface wettability on the single-bubble dynamics. We employed the dynamic van der Waals theory [Phys. Rev. E 75, 036304 (2007)], a diffuse-interface model for liquid-vapor flows involving liquid-vapor transition in nonuniform temperature fields. We first perform simulations for bubbles on homogeneous surfaces. We find that an increase in either the contact angle or the surface superheating can enhance the bubble spreading over the heating surface and increase the bubble departure diameter as well and therefore facilitate the transition into film boiling. We then examine the dynamics of bubbles on patterned surfaces, which incorporate the advantages of both hydrophobic and hydrophilic surfaces. The central hydrophobic region increases the thermodynamic probability of bubble nucleation while the surrounding hydrophilic region hinders the continuous bubble spreading by pinning the contact line at the hydrophobic-hydrophilic intersection. This leads to a small bubble departure diameter and therefore prevents the transition from nucleate boiling into film boiling. With the bubble nucleation probability increased and the bubble departure facilitated, the efficiency of heat transfer on such patterned surfaces is highly enhanced, as observed experimentally [Int. J. Heat Mass Transfer 57, 733 (2013)]. In addition, the stick-slip motion of contact line on patterned surfaces is demonstrated in one-component fluids, with the effect weakened by surface superheating.

  14. Insulation of Nitrocellulose Boiling Tubs at Radford Army Ammunition Plant

    DTIC Science & Technology

    1982-03-01

    control system. The amount of steam usea for the on-boil cycle with the single-sensor autocontrol averaged 647 kg/hr (1426 lb/hr) (test 1, table 2...This was a reduc- tion of 210 kg/hr (463 lb/hr) over the manually controlled uninsulated tub. Steam usage with the single sensor autocontrol and...uninsulated tub. At times durin)g the on- boil cycle of tests I and 2, the temperature of the manual sensor was different from the autocontrol sensor indicating

  15. Surfactant effects on interfacial flow and thermal transport processes during phase change in film boiling

    NASA Astrophysics Data System (ADS)

    Premnath, Kannan N.; Hajabdollahi, Farzaneh; Welch, Samuel W. J.

    2018-04-01

    The presence of surfactants in two-phase flows results in the transport and adsorption of surfactants to the interface, and the resulting local interfacial concentration significantly influences the surface tension between the liquid and vapor phases in a fluid undergoing phase change. This computational study is aimed at understanding and elucidating the mechanisms of enhanced flows and thermal transport processes in film boiling due to the addition of surfactants. A change in surface tension results in a change in the critical Rayleigh-Taylor wavelength leading to different bubble release patterns and a change in the overall heat transfer rates. Due to the presence of surfactants, an additional transport mechanism of the Marangoni convection arises from the resulting tangential gradients in the surfactant concentration along the phase interface. Our computational approach to study such phenomena consists of representing the interfacial motion by means of the coupled level set-volume-of-fluid method, the fluid motion via the classical marker-and-cell approach, as well as representations for the bulk transport of energy and surfactants, in conjunction with a phase change model and an interfacial surfactant model. Using such an approach, we perform numerical simulations of surfactant-laden single mode as well as multiple mode film boiling and study the effect of surfactants on the transport processes in film boiling, including bubble release patterns, vapor generation rates, and heat transfer rates at different surfactant concentrations. The details of the underlying mechanisms will be investigated and interpreted.

  16. Experimental Investigation of Pool Boiling Heat Transfer Enhancement in Microgravity in the Presence of Electric Fields

    NASA Technical Reports Server (NTRS)

    Herman, Cila

    1999-01-01

    test cell was developed. All four vertical walls of the test cell are transparent, and they allow transillumination with laser light for visualization experiments by HI. The bottom electrode is a copper cylinder, which is electrically grounded. The copper block is heated with a resistive heater and it is equipped with 6 thermocouples that provide reference temperatures for the measurements with HI. The top electrode is a mesh electrode. Bubbles are injected with a syringe into the test cell through the bottom electrode. The working fluids presently used in the interferometric visualization experiments, water and PF 5052, satisfy requirements regarding thermophysical, optical and electrical properties. A 30kV power supply equipped with a voltmeter allows to apply the electric field to the electrodes during the experiments. The magnitude of the applied voltage can be adjusted either manually or through the LabVIEW data acquisition and control system connected to a PC. Temperatures of the heated block are recorded using type-T thermocouples, whose output is read by a data acquisition system. Images of the bubbles are recorded with 35mm photographic and 16mm high-speed cameras, scanned and analyzed using various software packages. Visualized temperature fields HI allows the visualization of temperature fields in the vicinity of bubbles during boiling in the form of fringes. Typical visualized temperature distributions around the air bubbles injected into the thermal boundary layer in PF5052 are shown. The temperature of the heated surface is 35 C. The temperature difference for a pair of fringes is approximately 0.05 C. The heat flux applied to the bottom surface is moderate, and the fringe patterns are regular. In the image a bubble penetrating the thermal boundary layer is visible. Because of the axial symmetry of the problem, simplified reconstruction techniques can be applied to recover the temperature field. The thermal plume developing above the heated surface for

  17. Prevotella jejuni sp. nov., isolated from the small intestine of a child with coeliac disease.

    PubMed

    Hedberg, Maria E; Israelsson, Anne; Moore, Edward R B; Svensson-Stadler, Liselott; Wai, Sun Nyunt; Pietz, Grzegorz; Sandström, Olof; Hernell, Olle; Hammarström, Marie-Louise; Hammarström, Sten

    2013-11-01

    Five obligately anaerobic, Gram-stain-negative, saccharolytic and proteolytic, non-spore-forming bacilli (strains CD3 : 27, CD3 : 28(T), CD3 : 33, CD3 : 32 and CD3 : 34) are described. All five strains were isolated from the small intestine of a female child with coeliac disease. Cells of the five strains were short rods or coccoid cells with longer filamentous forms seen sporadically. The organisms produced acetic acid and succinic acid as major metabolic end products. Phylogenetic analysis based on comparative 16S rRNA gene sequence analysis revealed close relationships between CD3 : 27, CD3 : 28(T) and CD3 : 33, between CD3 : 32 and Prevotella histicola CCUG 55407(T), and between CD3 : 34 and Prevotella melaninogenica CCUG 4944B(T). Strains CD3 : 27, CD3 : 28(T) and CD3 : 33 were clearly different from all recognized species within the genus Prevotella and related most closely to but distinct from P. melaninogenica. Based on 16S rRNA, RNA polymerase β-subunit (rpoB) and 60 kDa chaperonin protein subunit (cpn60) gene sequencing, and phenotypic, chemical and biochemical properties, strains CD3 : 27, CD3 : 28(T) and CD3 : 33 are considered to represent a novel species within the genus Prevotella, for which the name Prevotella jejuni sp. nov. is proposed. Strain CD3 : 28(T) ( = CCUG 60371(T) = DSM 26989(T)) is the type strain of the proposed novel species. All five strains were able to form homologous aggregates, in which tube-like structures were connecting individual bacteria cells. The five strains were able to bind to human intestinal carcinoma cell lines at 37 °C.

  18. Prevotella jejuni sp. nov., isolated from the small intestine of a child with coeliac disease

    PubMed Central

    Israelsson, Anne; Moore, Edward R. B.; Svensson-Stadler, Liselott; Wai, Sun Nyunt; Pietz, Grzegorz; Sandström, Olof; Hernell, Olle; Hammarström, Marie-Louise

    2013-01-01

    Five obligately anaerobic, Gram-stain-negative, saccharolytic and proteolytic, non-spore-forming bacilli (strains CD3 : 27, CD3 : 28T, CD3 : 33, CD3 : 32 and CD3 : 34) are described. All five strains were isolated from the small intestine of a female child with coeliac disease. Cells of the five strains were short rods or coccoid cells with longer filamentous forms seen sporadically. The organisms produced acetic acid and succinic acid as major metabolic end products. Phylogenetic analysis based on comparative 16S rRNA gene sequence analysis revealed close relationships between CD3 : 27, CD3 : 28T and CD3 : 33, between CD3 : 32 and Prevotella histicola CCUG 55407T, and between CD3 : 34 and Prevotella melaninogenica CCUG 4944BT. Strains CD3 : 27, CD3 : 28T and CD3 : 33 were clearly different from all recognized species within the genus Prevotella and related most closely to but distinct from P. melaninogenica. Based on 16S rRNA, RNA polymerase β-subunit (rpoB) and 60 kDa chaperonin protein subunit (cpn60) gene sequencing, and phenotypic, chemical and biochemical properties, strains CD3 : 27, CD3 : 28T and CD3 : 33 are considered to represent a novel species within the genus Prevotella, for which the name Prevotella jejuni sp. nov. is proposed. Strain CD3 : 28T ( = CCUG 60371T = DSM 26989T) is the type strain of the proposed novel species. All five strains were able to form homologous aggregates, in which tube-like structures were connecting individual bacteria cells. The five strains were able to bind to human intestinal carcinoma cell lines at 37 °C. PMID:23793857

  19. Stable Concentrated Emulsions of the 1-Monoglyceride of Capric Acid (Monocaprin) with Microbicidal Activities against the Food-Borne Bacteria Campylobacter jejuni, Salmonella spp., and Escherichia coli

    PubMed Central

    Thormar, Halldor; Hilmarsson, Hilmar; Bergsson, Gudmundur

    2006-01-01

    Of 11 fatty acids and monoglycerides tested against Campylobacter jejuni, the 1-monoglyceride of capric acid (monocaprin) was the most active in killing the bacterium. Various monocaprin-in-water emulsions were prepared which were stable after storage at room temperature for many months and which retained their microbicidal activity. A procedure was developed to manufacture up to 500 ml of 200 mM preconcentrated emulsions of monocaprin in tap water. The concentrates were clear and remained stable for at least 12 months. They were active against C. jejuni upon 160- to 200-fold dilution in tap water and caused a >6- to 7-log10 reduction in viable bacterial count in 1 min at room temperature. The addition of 0.8% Tween 40 to the concentrates as an emulsifying agent did not change the microbicidal activity. Emulsions of monocaprin killed a variety of Campylobacter isolates from humans and poultry and also killed strains of Campylobacter coli and Campylobacter lari, indicating a broad anticampylobacter activity. Emulsions of 1.25 mM monocaprin in citrate-lactate buffer at pH 4 to 5 caused a >6- to 7-log10 reduction in viable bacterial counts of Salmonella spp. and Escherichia coli in 10 min. C. jejuni was also more susceptible to monocaprin emulsions at low pH. The addition of 5 and 10 mM monocaprin emulsions to Campylobacter-spiked chicken feed significantly reduced the bacterial contamination. These results are discussed in view of the possible utilization of monocaprin emulsions in controlling the spread of food-borne bacteria from poultry to humans. PMID:16391087

  20. Bubble Dynamics, Two-Phase Flow, and Boiling Heat Transfer in Microgravity

    NASA Technical Reports Server (NTRS)

    Chung, Jacob N.

    1996-01-01

    The objective of the research is to study the feasibility of employing an external force to replace the buoyancy force in order to maintain nucleate boiling in microgravity. We have found that a bulk velocity field, an electric field and an acoustic field could each play the role of the gravity field in microgravity. Nucleate boiling could be maintained by any one of the three external force fields in space.