Science.gov

Sample records for campylobacter jejuni adherence

  1. Campylobacter jejuni: components for adherence to and invasion of eukaryotic cells.

    PubMed

    Lugert, Raimond; Gross, Uwe; Zautner, Andreas E

    2015-01-01

    Campylobacter (C.) jejuni is the most important reported cause for bacterial diarrhoea. The infection can be accompanied by fever and abdominal cramps and in rare cases the Guillain-Barré syndrome or reactive arthritis can develop as a post-infection complication. Several biological properties of Cjejuni, e. g. motility and chemotaxis, contribute to the biological fitness of the pathogen. For this, deficiencies in the function of these features clearly reduce the pathogenicity of C. jejuni without being a virulence factor per se. Opposing to this, there are two essential requirements to determine the virulence of C. jejuni which represent the adherence to, and the invasion of host cells. Thereby, adherence, as a virulence factor, is mediated by many different bacterial-derived components like proteins but also by several oligo- and polysaccharide structures that are linked to surface proteins but also to the flagella. In addition, several invasion-relevant features of C. jejuni have been detected so far. Whereas some of them are described functionally to modulate cytoskeleton arrangement of the host cell, others are only described as invasion relevant. Indeed, investigations with respect to the pathogenic potential of some adherence- and invasion-relevant components did not give identical results indicating that their relevance might depend on the interplay of the respective C. jejuni strains used in these studies with the corresponding host cells. This review summarizes the C. jejuni components for adherence to and invasion of host cells together with their particular mode of action if known.

  2. Campylobacter jejuni organism (image)

    MedlinePlus

    Campylobacter jejuni infection causes cramping, diarrhea, abdominal pain and fever within 2 to 5 days after a person has been exposed to the organism. Campylobacter jejuni is one of the most common bacterial causes ...

  3. 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.

  4. 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. PMID:26219363

  5. Campylobacter jejuni, other campylobacters

    Technology Transfer Automated Retrieval System (TEKTRAN)

    For approximately three decades, the genus Campylobacter has had increased focus as a threat to food safety, due to the rise in enteritis in humans caused by consumption or handling of foods contaminated with the organism. For this reason, numerous research studies have been conducted and books wri...

  6. The hyperosmotic stress response of Campylobacter jejuni

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The hyperosmotic stress response of Campylobacter jejuni: The diarrheal pathogen Campylobacter jejuni and other gastrointestinal bacteria encounter changes in osmolarity in the environment, through exposure to food processing, or upon entering host organisms, where osmotic adaptation can be associa...

  7. Resistance mechanisms in Campylobacter jejuni.

    PubMed

    Iovine, Nicole M

    2013-04-01

    Campylobacter jejuni is a major cause of food-borne gastroenteritis worldwide. While mortality is low, morbidity imparted by post-infectious sequelae such as Guillain-Barré syndrome, Reiter syndrome/reactive arthritis and irritable bowel syndrome is significant. In addition, the economic cost is high due to lost productivity. Food animals, particularly poultry, are the main reservoirs of C. jejuni. The over-use of antibiotics in the human population and in animal husbandry has led to an increase in antibiotic-resistant infections, particularly with fluoroquinolones. This is problematic because C. jejuni gastroenteritis is clinically indistinguishable from that caused by other bacterial pathogens, and such illnesses are usually treated empirically with fluoroquinolones. Since C. jejuni is naturally transformable, acquisition of additional genes imparting antibiotic resistance is likely. Therefore, an understanding of the antibiotic resistance mechanisms in C. jejuni is needed to provide proper therapy both to the veterinary and human populations.

  8. Oxygen requirement and tolerance of Campylobacter jejuni.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The human pathogen Campylobacter jejuni is considered a microaerophile, yet it has been shown to grow in vitro under atmospheres with elevated oxygen tensions. Hence, a better understanding of the oxygen requirement and tolerance of C. jejuni is required. Bacterial growth was measured under various ...

  9. [Selected mechanisms of pathogenicity of Campylobacter jejuni].

    PubMed

    Rokosz-Chudziak, Natalia; Rastawicki, Waldemar

    2014-01-01

    Campylobacter jejuni is one of the most common causes of bacterial diarrhea worldwide. Various virulence factors of C. jejuni contribute to survival and participate in the induction of infection in the human body. The virulence mechanisms such as motility, toxin production or invasive properties allow the bacteria to survive in the human body. The article presents the current knowledge on selected virulence mechanisms of C. jejuni.

  10. Isolation of Campylobacter jejuni from groundwater.

    PubMed

    Stanley, K; Cunningham, R; Jones, K

    1998-07-01

    A pollution event which occurred at a spring in the Arnside area of Cumbria provided an opportunity to investigate whether Campylobacter jejuni could be detected in groundwater. Hydrological evidence suggested that the source of contamination was a dairy farm situated within the hydrological catchment of the polluted spring. The microbiological quality of the polluted spring was monitored during intervals over the following 12 months and compared with others in the area. Campylobacter jejuni was isolated by filter enrichment of 500 ml and 100 ml filtered volumes of groundwater. It was not isolated in the absence of faecal indicator species. Some strains of Camp. jejuni from water had identical biotypes to strains isolated from the dairy herd. This paper reports the first isolation of Camp. jejuni from groundwater using cultural methods and supports the theory that groundwater may be a vehicle for Campylobacter transmission.

  11. 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

  12. Campylobacter jejuni--an emerging foodborne pathogen.

    PubMed Central

    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. PMID:10081669

  13. 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

  14. Inhibition by pectic oligosaccharides of the invasion of undifferentiated and differentiated Caco-2 cells by Campylobacter jejuni

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The ability of pectic oligosaccharides (POS) to inhibit adherence to and invasion of undifferentiated (UC) and differentiated (DC) Caco-2 cells by Campylobacter jejuni (C. jejuni) was investigated. It was observed that both adherence and invasion were significantly higher in UC than in DC. POS (2.5 ...

  15. Campylobacter jejuni peritonitis complicating continuous ambulatory peritoneal dialysis.

    PubMed Central

    Pepersack, F; D'Haene, M; Toussaint, C; Schoutens, E

    1982-01-01

    We report the occurrence of Campylobacter jejuni peritonitis complicating C. jejuni enteritis in a patient treated with continuous ambulatory peritoneal dialysis. Cure followed oral administration of erythromycin and intraperitoneal therapy with gentamicin. PMID:7153322

  16. Campylobacter jejuni in broilers: the role of vertical transmission.

    PubMed Central

    Shanker, S.; Lee, A.; Sorrell, T. C.

    1986-01-01

    The role of broiler eggs in the transmission of Campylobacter jejuni to broiler grow-out flocks was investigated. Six breeder flocks supplying broiler eggs to hatcheries were examined for cloacal carriage of C. jejuni. Of 240 birds tested, 178 (74%) were C. jejuni-positive. Eggs from these birds examined for C. jejuni penetration of the egg shell indicated that 185 of 187 were campylobacter-free. Eggs from breeder flocks of unknown C. jejuni status were also examined for C. jejuni shell penetration. C. jejuni was not isolated from 142 eggs examined. A further 193 hatchery eggs incubated and hatched in the laboratory were campylobacter-free. Six farms containing the progeny of C. jejuni-positive breeder flocks were monitored. Eight hundred and forty birds from 14 flocks in these grow-out farms were campylobacter-free during their 6-week grow-out period. Experimental egg-penetration studies indicated that C. jejuni transmission via the egg is not easily effected. Of 257 eggs surface-challenged with C. jejuni, 162 hatched; all were campylobacter-free. Of 167 eggs injected with C. jejuni, 12 hatched; 2 of these were colonized with C. jejuni. Our data do not support a role for vertical transmission of C. jejuni in commercial broiler production. PMID:3701037

  17. Complete Genomic Sequence of Campylobacter jejuni subsp. jejuni HS:19 Strain RM1285 Isolated from Packaged Chicken

    PubMed Central

    Huynh, Steven; Heikema, Astrid P.

    2016-01-01

    Poultry products serve as the main source of Campylobacter jejuni subsp. jejuni infections in humans. C. jejuni subsp. jejuni infections are a leading cause of foodborne gastroenteritis and are a prevalent antecedent to Guillain-Barré syndrome. This study describes the genome of C. jejuni subsp. jejuni HS:19 strain RM1285, isolated from packaged chicken in California. PMID:27795263

  18. Post-genome Analysis of the Foodborne Pathogen Campylobacter jejuni

    NASA Astrophysics Data System (ADS)

    Kay, Emily J.; Gundogdu, Ozan; Wren, Brendan

    The human pathogen Campylobacter jejuni is part of the genus Campylobacter that lies within the epsilon proteobacteria subclass of bacteria. The nearest family in phylogenetic terms is the Helicobacteraceae which includes the Helicobacter and Wolinella genuses. Campylobacter species are Gram-negative, curved rod shaped or spiral and are motile (via polar flagella).

  19. Campylobacter jejuni non-culturable coccoid cells.

    PubMed

    Beumer, R R; de Vries, J; Rombouts, F M

    1992-01-01

    The behaviour of Campylobacter jejuni in the environment is poorly documented. Rapid loss of viability on culture media is reported. This phenomenon is associated with the development of so-called coccoid cells. It has been suggested that these cells can be infective to animals and man. Results obtained with ATP-measurements of coccoid cells and Direct Viable Count (DVC) support this hypothesis. Introduction of coccoid cells into simulated gastric, ileal and colon environments did not result in the presence of culturable cells. Oral administration to laboratory animals and volunteers caused no typical symptoms of campylobacteriosis. Until 30 days after uptake of the cells antibodies against C. jejuni could not be detected in the blood, and the presence of this microorganism in stool samples could not be demonstrated.

  20. Substrate utilization by Campylobacter jejuni and Campylobacter coli

    SciTech Connect

    Westfall, H.N.; Rollins, D.M.; Weiss, E.

    1986-10-01

    An attempt was made to elucidate in Campylobacter spp. some of the physiologic characteristics that are reflected in the kinetics of CO/sub 2/ formation from four /sup 14/C-labeled substrates. Campylobacter jejuni and C. coli were grown in a biphasic medium, and highly motile spiral cells were harvested at 12 h. The cells were incubated with 0.02 M glutamate, glutamine, ..cap alpha..-ketoglutarate, or formate, or with concentrations of these substrates ranging from 0.0032 to 0.125 M. All four substrates were metabolized very rapidly by both species. A feature of many of these reactions, particularly obvious with /sup 2/chemically bond-ketoglutarate, was an immediate burst of CO/sub 2/ production followed by CO/sub 2/ evolution at a more moderate rate. These diphasic kinetics of substrate utilization were not seen in comparable experiments with Escherichia coli grown and tested under identical conditions. With C. jejuni, CO/sub 2/ production from formate proceeded rapidly for the entire period of incubation. The rate of metabolism of glutamate, glutamine, and ..cap alpha..-ketoglutarate by both species was greatly enhanced by increased substrate concentration. The approach to the study of the metabolism of campylobacters here described may be useful in detecting subtle changes in the physiology of cells as they are maintained past their logarithmic growth phase.

  1. 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

  2. Presence of antibodies against campylobacter flagellar capping proteins versus campylobacter jejuni isolation in broilers

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Campylobacter jejuni is the leading foodborne pathogen that causes human acute bacterial gastroenteritis worldwide. Human cases have been linked to consumption and/or handling of contaminated poultry products. Although Campylobacter jejuni is commonly regarded as a commensal in broiler cecal micro...

  3. Effects of disinfectants on Campylobacter jejuni.

    PubMed Central

    Wang, W L; Powers, B W; Leuchtefeld, N W; Blaser, M J

    1983-01-01

    Because of the increasing recognition that Campylobacter jejuni is an important enteric pathogen of humans, we studied the effects of widely used disinfectants on the viability of this organism. At an inoculum size of 10(3) to 10(4) CFU/ml, 1.25 mg of hypochlorite per liter killed three strains within 1 min. At an inoculum size of 10(6) to 10(7) CFU/ml, 5 mg of hypochlorite per liter killed three strains within 15 min. Killing of similar concentrations of C. jejuni and Escherichia coli by hypochlorite was approximately the same. At the high inoculum, 0.15% phenolic compound, 10 mg of iodophor per liter, 1:50,000 quaternary ammonium compound, 70% ethyl alcohol, and 0.125% glutaraldehyde killed all three strains within 1 min. These studies demonstrate that, under the conditions we tested (pH 7.0; 24 to 26 degrees C), the recommended standard concentrations of disinfecting agents are adequate to destroy C. jejuni. PMID:6305266

  4. The heat-sensitivity of Campylobacter jejuni in milk.

    PubMed Central

    Waterman, S. C.

    1982-01-01

    It is now established that milk can be a vehicle for the spread of enteritis due to Campylobacter jejuni. By determining the lethal effect of heat on six isolates it has been shown that C. jejuni is unlikely to survive pasteurization. PMID:7086118

  5. Identification of possible virulence marker from Campylobacter jejuni isolates.

    PubMed

    Harrison, James W; Dung, Tran Thi Ngoc; Siddiqui, Fariha; Korbrisate, Sunee; Bukhari, Habib; Tra, My Phan Vu; Hoang, Nguyen Van Minh; Carrique-Mas, Juan; Bryant, Juliet; Campbell, James I; Studholme, David J; Wren, Brendan W; Baker, Stephen; Titball, Richard W; Champion, Olivia L

    2014-06-01

    A novel protein translocation system, the type-6 secretion system (T6SS), may play a role in virulence of Campylobacter jejuni. We investigated 181 C. jejuni isolates from humans, chickens, and environmental sources in Vietnam, Thailand, Pakistan, and the United Kingdom for T6SS. The marker was most prevalent in human and chicken isolates from Vietnam.

  6. Colonization of broilers by Campylobacter jejuni internalized within Acanthamoeba castellanii

    Technology Transfer Automated Retrieval System (TEKTRAN)

    We present the first report that Campylobacter jejuni, internalized within Acanthamoeba castellanii, colonized broilers. After 1, 3, 7 and 14 days post challenge none of the broilers challenged with negative controls were colonized, but were with internalized C. jejuni. The biology of protozoa-Cam...

  7. Detection and characterization of autoagglutination activity by Campylobacter jejuni.

    PubMed

    Misawa, N; Blaser, M J

    2000-11-01

    In several gram-negative bacterial pathogens, autoagglutination (AAG) activity is a marker for interaction with host cells and virulence. Campylobacter jejuni strains also show AAG, but this property varies considerably among strains. To examine the characteristics of C. jejuni AAG, we developed a quantitative in vitro assay. For strain 81-176, which shows high AAG, activity was optimal for cells grown for < or = 24 h, was independent of growth temperature, and was best measured for cells suspended in phosphate-buffered saline at 25 degrees C for 24 h. AAG activity was heat labile and was abolished by pronase or acid-glycine (pH 2.2) treatment but not by lipase, DNase, or sodium metaperiodate. Strain 4182 has low AAG activity, but extraction with water increased AAG, suggesting the loss of an inhibitor. Strain 6960 has weak AAG with no effect due to water extraction. Our study with clinical isolates suggests that C. jejuni strains may be grouped into three AAG phenotypes. A variant derived from strain 81116 that is flagellate but immotile showed the strong AAG exhibited by the parent strain, suggesting that motility per se is not necessary for the AAG activity. AAG correlated with both bacterial hydrophobicity and adherence to INT407 cells. Mutants which lack flagella (flaA, flaB, and flbA) or common cell surface antigen (peb1A) were constructed in strain 81-176 by natural transformation-mediated allelic exchange. Both AAG activity and bacterial hydrophobicity were abolished in the aflagellate mutants but not the peb1A mutant. In total, these findings indicate that C. jejuni AAG is highly associated with flagellar expression.

  8. Conformational analysis of the Campylobacter jejuni porin.

    PubMed Central

    Bolla, J M; Loret, E; Zalewski, M; Pagés, J M

    1995-01-01

    The major outer membrane protein (MOMP) of Campylobacter jejuni was purified to homogeneity by selective solubilization and fast protein liquid chromatography. The amino acid composition of the MOMP indicates the presence of cysteine residues. The amino-terminal sequence, determined over 31 residues, shows no significant homology with any other porin from gram-negative bacteria except in a discrete region. Immunocross-reactivity between Escherichia coli OmpC and the MOMP was analyzed, and a common antigenic site between these two porins was identified with an anti-peptide antibody. From circular dichroism and immunological investigations, the existence of a stable folded monomer, containing a high level of beta-sheet secondary structure, is evident. Conformational analyses show the presence of a native trimeric state generated by association of the three folded monomers; the stability of this trimer is reduced compared with that of E. coli porins. This study clearly reveals that the C. jejuni MOMP is related to the family of trimeric bacterial porins. PMID:7543469

  9. Specificity of monoclonal antibodies to Campylobacter jejuni lipopolysaccharide antigens.

    PubMed

    Brooks, B W; Mihowich, J G; Blais, B W; Yamazaki, H

    1998-01-01

    Monoclonal antibodies (Mabs) were produced to the lipopolysaccharide antigens of Campylobacter jejuni strain 1249 (Penner serotype O:2/63). A polymyxin-cloth based enzyme immunoassay (pCEIA) was used for initial screening and for evaluating the specificity of these antibodies. Seven Mabs reacted with at least 11 and as many as 14 of 15 C. jejuni strains (representing 8 different Penner serotypes). These seven Mabs did not cross-react with any of 16 non-Campylobacter bacteria commonly encountered in food, with only two exceptions. Several combinations of these Mabs in pairs reacted with all 15 C. jejuni strains. These results suggest that pCEIA employing two of these Mabs in combination is potentially useful for detection of Campylobacter jejuni in foods and other samples.

  10. Quantitative Proteomics of Intracellular Campylobacter jejuni Reveals Metabolic Reprogramming

    PubMed Central

    Liu, Xiaoyun; Gao, Beile; Novik, Veronica; Galán, Jorge E.

    2012-01-01

    Campylobacter jejuni is the major cause of bacterial food-borne illness in the USA and Europe. An important virulence attribute of this bacterial pathogen is its ability to enter and survive within host cells. Here we show through a quantitative proteomic analysis that upon entry into host cells, C. jejuni undergoes a significant metabolic downshift. Furthermore, our results indicate that intracellular C. jejuni reprograms its respiration, favoring the respiration of fumarate. These results explain the poor ability of C. jejuni obtained from infected cells to grow under standard laboratory conditions and provide the bases for the development of novel anti microbial strategies that would target relevant metabolic pathways. PMID:22412372

  11. 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.

  12. Effect of enteroviruses on adherence to and invasion of HEp-2 cells by Campylobacter isolates.

    PubMed Central

    Konkel, M E; Joens, L A

    1990-01-01

    Coinfection of HEp-2 epithelial cells with coxsackievirus B3, echovirus 7, poliovirus (LSc type 1), porcine enterovirus, and Campylobacter isolates was performed to determine if a synergistic effect could be obtained. The invasiveness of Campylobacter jejuni ATCC 33560 was significantly increased for HEp-2 cells preinfected with echovirus 7, coxsackievirus B3, and UV-inactivated (noninfectious) coxsackievirus B3 particles. Additionally, the invasiveness of C. jejuni M96, a clinical isolate, was significantly increased for HEp-2 cells preinfected with coxsackievirus B3. Poliovirus and porcine enterovirus had no effect on C. jejuni ATCC 33560 adherence and invasiveness. Furthermore, poliovirus had no effect on the ability of C. jejuni M96 to adhere to and invade HEp-2 cells. Campylobacter hyointestinalis and Campylobacter mucosalis, two noninvasive isolates, did not invade virus-infected HEp-2 cells. The increase in the invasiveness of C. jejuni appeared to be the result of specific interactions between the virus and the HEp-2 cell membrane. The data suggest that the invasiveness of Campylobacter spp. is dependent upon the inherent properties of the organism. Virus-induced cell alterations can potentiate the invasiveness of virulent Campylobacter spp. but are not sufficient to allow internalization of noninvasive bacteria. PMID:2156779

  13. Antibiotic susceptibility of clinical isolates of Campylobacter jejuni and Campylobacter coli in Ontario, Canada during 2011-2013.

    PubMed

    Riley, Andrew; Eshaghi, Alireza; Olsha, Romy; Allen, Vanessa G; Patel, Samir N

    2015-11-01

    A total of 219 clinical isolates of Campylobacter spp. including 180 Campylobacter jejuni and 39 Campylobacter coli were assessed for in vitro antimicrobial susceptibility. Resistance among C. coli was higher for ciprofloxacin (41% versus 30.80%), erythromycin (12.80% versus 3.90%) and lower for tetracycline (53.80% versus 64.60%) compared to C. jejuni.

  14. Host adaption to the bacteriophage carrier state of Campylobacter jejuni.

    PubMed

    Brathwaite, Kelly J; Siringan, Patcharin; Connerton, Phillippa L; Connerton, Ian F

    2015-01-01

    The carrier state of the foodborne pathogen Campylobacter jejuni represents an alternative life cycle whereby virulent bacteriophages can persist in association with host bacteria without commitment to lysogeny. Host bacteria exhibit significant phenotypic changes that improve their ability to survive extra-intestinal environments, but exhibit growth-phase-dependent impairment in motility. We demonstrate that early exponential phase cultures become synchronised with respect to the non-motile phenotype, which corresponds with a reduction in their ability to adhere to and invade intestinal epithelial cells. Comparative transcriptome analyses (RNA-seq) identify changes in gene expression that account for the observed phenotypes: downregulation of stress response genes hrcA, hspR and per and downregulation of the major flagellin flaA with the chemotactic response signalling genes cheV, cheA and cheW. These changes present mechanisms by which the host and bacteriophage can remain associated without lysis, and the cultures survive extra-intestinal transit. These data provide a basis for understanding a critical link in the ecology of the Campylobacter bacteriophage.

  15. Host adaption to the bacteriophage carrier state of Campylobacter jejuni

    PubMed Central

    Brathwaite, Kelly J.; Siringan, Patcharin; Connerton, Phillippa L.; Connerton, Ian F.

    2015-01-01

    The carrier state of the foodborne pathogen Campylobacter jejuni represents an alternative life cycle whereby virulent bacteriophages can persist in association with host bacteria without commitment to lysogeny. Host bacteria exhibit significant phenotypic changes that improve their ability to survive extra-intestinal environments, but exhibit growth-phase-dependent impairment in motility. We demonstrate that early exponential phase cultures become synchronised with respect to the non-motile phenotype, which corresponds with a reduction in their ability to adhere to and invade intestinal epithelial cells. Comparative transcriptome analyses (RNA-seq) identify changes in gene expression that account for the observed phenotypes: downregulation of stress response genes hrcA, hspR and per and downregulation of the major flagellin flaA with the chemotactic response signalling genes cheV, cheA and cheW. These changes present mechanisms by which the host and bacteriophage can remain associated without lysis, and the cultures survive extra-intestinal transit. These data provide a basis for understanding a critical link in the ecology of the Campylobacter bacteriophage. PMID:26004283

  16. Reducing Campylobacter jejuni colonization of poultry via vaccination.

    PubMed

    Neal-McKinney, Jason M; Samuelson, Derrick R; Eucker, Tyson P; Nissen, Mark S; Crespo, Rocio; Konkel, Michael E

    2014-01-01

    Campylobacter jejuni is a leading bacterial cause of human gastrointestinal disease worldwide. While C. jejuni is a commensal organism in chickens, case-studies have demonstrated a link between infection with C. jejuni and the consumption of foods that have been cross-contaminated with raw or undercooked poultry. We hypothesized that vaccination of chickens with C. jejuni surface-exposed colonization proteins (SECPs) would reduce the ability of C. jejuni to colonize chickens, thereby reducing the contamination of poultry products at the retail level and potentially providing a safer food product for consumers. To test our hypothesis, we injected chickens with recombinant C. jejuni peptides from CadF, FlaA, FlpA, CmeC, and a CadF-FlaA-FlpA fusion protein. Seven days following challenge, chickens were necropsied and cecal contents were serially diluted and plated to determine the number of C. jejuni per gram of material. The sera from the chickens were also analyzed to determine the concentration and specificity of antibodies reactive against the C. jejuni SECPs. Vaccination of chickens with the CadF, FlaA, and FlpA peptides resulted in a reduction in the number of C. jejuni in the ceca compared to the non-vaccinated C. jejuni-challenged group. The greatest reduction in C. jejuni colonization was observed in chickens injected with the FlaA, FlpA, or CadF-FlaA-FlpA fusion proteins. Vaccination of chickens with different SECPs resulted in the production of C. jejuni-specific IgY antibodies. In summary, we show that the vaccination of poultry with individual C. jejuni SECPs or a combination of SECPs provides protection of chickens from C. jejuni colonization.

  17. Adaptive mechanisms of Campylobacter jejuni to erythromycin treatment

    PubMed Central

    2013-01-01

    Background Macrolide is the drug of choice to treat human campylobacteriosis, but Campylobacter resistance to this antibiotic is rising. The mechanisms employed by Campylobacter jejuni to adapt to erythromycin treatment remain unknown and are examined in this study. The transcriptomic response of C. jejuni NCTC 11168 to erythromycin (Ery) treatment was determined by competitive microarray hybridizations. Representative genes identified to be differentially expressed were further characterized by constructing mutants and assessing their involvement in antimicrobial susceptibility, oxidative stress tolerance, and chicken colonization. Results Following the treatment with an inhibitory dose of Ery, 139 genes were up-regulated and 119 were down-regulated. Many genes associated with flagellar biosynthesis and motility was up-regulated, while many genes involved in tricarboxylic acid cycle, electron transport, and ribonucleotide biosynthesis were down-regulated. Exposure to a sub-inhibitory dose of Ery resulted in differential expression of much fewer genes. Interestingly, two putative drug efflux operons (cj0309c-cj0310c and cj1173-cj1174) were up-regulated. Although mutation of the two operons did not alter the susceptibility of C. jejuni to antimicrobials, it reduced Campylobacter growth under high-level oxygen. Another notable finding is the consistent up-regulation of cj1169c-cj1170c, of which cj1170c encodes a known phosphokinase, an important regulatory protein in C. jejuni. Mutation of the cj1169c-cj1170c rendered C. jejuni less tolerant to atmospheric oxygen and reduced Campylobacter colonization and transmission in chickens. Conclusions These findings indicate that Ery treatment elicits a range of changes in C. jejuni transcriptome and affects the expression of genes important for in vitro and in vivo adaptation. Up-regulation of motility and down-regulation of energy metabolism likely facilitate Campylobacter to survive during Ery treatment. These findings

  18. Genotypes and antibiotic resistance of canine Campylobacter jejuni isolates.

    PubMed

    Amar, Chantal; Kittl, Sonja; Spreng, David; Thomann, Andreas; Korczak, Bożena M; Burnens, André P; Kuhnert, Peter

    2014-01-10

    Campylobacter jejuni is the most important cause of bacterial gastroenteritis in humans. It is a commensal in many wild and domestic animals, including dogs. Whereas genotypes of human and chicken C. jejuni isolates have been described in some detail, only little information on canine C. jejuni genotypes is available. To gain more information on genotypes of canine C. jejuni and their zoonotic potential, isolates from routine diagnostics of diarrheic dogs as well as isolates of a prevalence study in non-diarrheic dogs were analyzed. Prevalence of thermophilic Campylobacter among non-diarrheic dogs was 6.3% for C. jejuni, 5.9% for Campylobacter upsaliensis and 0.7% for Campylobacter coli. The C. jejuni isolates were genotyped by multi locus sequence typing (MLST) and flaB typing. Resistance to macrolides and quinolones was genetically determined in parallel. Within the 134 genotyped C. jejuni isolates 57 different sequence types (ST) were found. Five STs were previously unrecognized. The most common STs were ST-48 (11.2%), ST-45 (10.5%) and ST-21 (6.0%). Whereas no macrolide resistance was found, 28 isolates (20.9%) were resistant to quinolones. ST-45 was significantly more prevalent in diarrheic than in non-diarrheic dogs. Within the common time frame of isolation 94% of the canine isolates had a ST that was also found in human clinical isolates. In conclusion, prevalence of C. jejuni in Swiss dogs is low but there is a large genetic overlap between dog and human isolates. Given the close contact between human and dogs, the latter should not be ignored as a potential source of human campylobacteriosis.

  19. Isolation of Campylobacter fetus subsp jejuni from zoo animals.

    PubMed

    Luechtefeld, N W; Cambre, R C; Wang, W L

    1981-12-01

    Over a 1-year period, 619 fecal specimens from animals at the Denver Zoo were cultured for Campylobacter fetus subsp jejuni. The organism was isolated from 35 animals, including 12 primates, 2 felids, a red panda, 13 hooved animals, 6 birds, and 1 reptile. Of 44 cultured fecal specimens from diarrheal animals, 31.8% were positive for Campylobacter, whereas only 5.6% of 575 specimens from animals without diarrhea were positive (P less than 0.001). Among 25 isolates tested, 12 serotypes were represented; several of these serotypes are commonly associated with Campylobacter enteritis in human beings. Campylobacter fetus subsp jejuni was isolated from 8% of 75 wild pigeons trapped on the zoo premises during winter months and from 26% of 75 trapped during March and April (P less than 0.01).

  20. Comparative characterization of the virulence gene clusters (lipooligosacharide [LOS] and capsular polysaccharide [CPS]) for Campylobacter coli, Campylobacter jejuni subsp. jejuni and related Campylobacter species

    PubMed Central

    Richards, Vincent P.; Lefébure, Tristan; Pavinski Bitar, Paulina D.; Stanhope, Michael J.

    2013-01-01

    Campylobacter jejuni subsp. jejuni and Campylobacter coli are leading causes of gastroenteritis, with virulence linked to cell surface carbohydrate diversity. Although the associated gene clusters are well studied for C. jejuni subsp. jejuni, C. coli has been largely neglected. Here we provide comparative analysis of the lipooligosacharide (LOS) and capsular polysaccharide (CPS) gene clusters, using genome and cluster sequence data for 36 C. coli strains, 67 C. jejuni subsp. jejuni strains and ten additional Campylobacter species. Similar to C. jejuni subsp. jejuni, C. coli showed high LOS/CPS gene diversity, with each cluster delineated into eight gene content classes. This diversity was predominantly due to extensive gene gain/loss, with the lateral transfer of genes likely occurring both within and between species and also between the LOS and CPS. Additional mechanisms responsible for LOS/CPS diversity included phase-variable homopolymeric repeats, gene duplication/inactivation, and possibly host environment selection pressure. Analyses also showed that (i) strains of C. coli and Campylobacter upsaliensis possessed genes homologous to the sialic acid genes implicated in the neurological disorder Guillain Barré syndrome (GBS), and (ii) C. coli LOS classes were differentiated between bovine and poultry hosts, potentially aiding post infection source tracking. PMID:23279811

  1. Comparative characterization of the virulence gene clusters (lipooligosaccharide [LOS] and capsular polysaccharide [CPS]) for Campylobacter coli, Campylobacter jejuni subsp. jejuni and related Campylobacter species.

    PubMed

    Richards, Vincent P; Lefébure, Tristan; Pavinski Bitar, Paulina D; Stanhope, Michael J

    2013-03-01

    Campylobacter jejuni subsp. jejuni and Campylobacter coli are leading causes of gastroenteritis, with virulence linked to cell surface carbohydrate diversity. Although the associated gene clusters are well studied for C. jejuni subsp. jejuni, C. coli has been largely neglected. Here we provide comparative analysis of the lipooligosaccharide (LOS) and capsular polysaccharide (CPS) gene clusters, using genome and cluster sequence data for 36 C. coli strains, 67 C. jejuni subsp. jejuni strains and ten additional Campylobacter species. Similar to C. jejuni subsp. jejuni, C. coli showed high LOS/CPS gene diversity, with each cluster delineated into eight gene content classes. This diversity was predominantly due to extensive gene gain/loss, with the lateral transfer of genes likely occurring both within and between species and also between the LOS and CPS. Additional mechanisms responsible for LOS/CPS diversity included phase-variable homopolymeric repeats, gene duplication/inactivation, and possibly host environment selection pressure. Analyses also showed that (i) strains of C. coli and Campylobacter upsaliensis possessed genes homologous to the sialic acid genes implicated in the neurological disorder Guillain-Barré syndrome (GBS), and (ii) C. coli LOS classes were differentiated between bovine and poultry hosts, potentially aiding post infection source tracking.

  2. Strain-specific probiotic (Lactobacillus helveticus) inhibition of Campylobacter jejuni invasion of human intestinal epithelial cells.

    PubMed

    Wine, Eytan; Gareau, Mélanie G; Johnson-Henry, Kathene; Sherman, Philip M

    2009-11-01

    Campylobacter jejuni is the most common bacterial cause of enterocolitis in humans, leading to diarrhoea and chronic extraintestinal diseases. Although probiotics are effective in preventing other enteric infections, beneficial microorganisms have not been extensively studied with C. jejuni. The aim of this study was to delineate the ability of selected probiotic Lactobacillus strains to reduce epithelial cell invasion by C. jejuni. Human colon T84 and embryonic intestine 407 epithelial cells were pretreated with Lactobacillus strains and then infected with two prototypic C. jejuni pathogens. Lactobacillus helveticus, strain R0052 reduced C. jejuni invasion into T84 cells by 35-41%, whereas Lactobacillus rhamnosus R0011 did not reduce pathogen invasion. Lactobacillus helveticus R0052 also decreased invasion of one C. jejuni isolate (strain 11168) into intestine 407 cells by 55%. Lactobacillus helveticus R0052 adhered to both epithelial cell types, which suggest that competitive exclusion could contribute to protection by probiotics. Taken together, these findings indicate that the ability of selected probiotics to prevent C. jejuni-mediated disease pathogenesis depends on the pathogen strain, probiotic strain and the epithelial cell type selected. The data support the concept of probiotic strain selectivity, which is dependent on the setting in which it is being evaluated and tested.

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

    PubMed Central

    Plickert, R.; Fischer, A.; Göbel, U. B.; Bereswill, S.

    2013-01-01

    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. PMID:24265916

  4. Inactivation of Campylobacter jejuni on poultry by ultraviolet light

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Campylobacter jejuni is a foodborne pathogen which is commonly associated with poultry, and is responsible for many foodborne illness outbreaks. Ultraviolet light (UV-C) is a U.S. Food and Drug Administration approved technology that can be used to treat foods and food contact surfaces. In this stud...

  5. Predominant Campylobacter jejuni Sequence Types Persist in Finnish Chicken Production

    PubMed Central

    Llarena, Ann-Katrin; Huneau, Adeline; Hakkinen, Marjaana; Hänninen, Marja-Liisa

    2015-01-01

    Consumption and handling of chicken meat are well-known risk factors for acquiring campylobacteriosis. This study aimed to describe the Campylobacter jejuni population in Finnish chickens and to investigate the distribution of C. jejuni genotypes on Finnish chicken farms over a period of several years. We included 89.8% of the total C. jejuni population recovered in Finnish poultry during 2004, 2006, 2007, 2008, and 2012 and used multilocus sequence typing (MLST) and pulsed-field gel electrophoresis (PFGE) to characterize the 380 isolates. The typing data was combined with isolate information on collection-time and farm of origin. The C. jejuni prevalence in chicken slaughter batches was low (mean 3.0%, CI95% [1.8%, 4.2%]), and approximately a quarter of Finnish chicken farms delivered at least one positive chicken batch yearly. In general, the C. jejuni population was diverse as represented by a total of 63 sequence types (ST), but certain predominant MLST lineages were identified. ST-45 clonal complex (CC) accounted for 53% of the isolates while ST-21 CC and ST-677 CC covered 11% and 9% of the isolates, respectively. Less than half of the Campylobacter positive farms (40.3%) delivered C. jejuni-contaminated batches in multiple years, but the genotypes (ST and PFGE types) generally varied from year to year. Therefore, no evidence for a persistent C. jejuni source for the colonization of Finnish chickens emerged. Finnish chicken farms are infrequently contaminated with C. jejuni compared to other European Union (EU) countries, making Finland a valuable model for further epidemiological studies of the C. jejuni in poultry flocks. PMID:25700264

  6. Screening of Australian plants for antimicrobial activity against Campylobacter jejuni.

    PubMed

    Kurekci, Cemil; Bishop-Hurley, Sharon L; Vercoe, Philip E; Durmic, Zoey; Al Jassim, Rafat A M; McSweeney, Christopher S

    2012-02-01

    Campylobacter jejuni is the most common cause of acute enteritis in humans, with symptoms such as diarrhoea, fever and abdominal cramps. In this study, 115 extracts from 109 Australian plant species were investigated for their antimicrobial activities against two C. jejuni strains using an in vitro broth microdilution assay. Among the plants tested, 107 (93%) extracts showed activity at a concentration between 32 and 1024 µg/mL against at least one C. jejuni strain. Seventeen plant extracts were selected for further testing against another six C. jejuni strains, as well as Campylobacter coli, Escherichia coli, Salmonella typhimurium, Bacillus cereus, Proteus mirabilis and Enterococcus faecalis. The extract from Eucalyptus occidentalis demonstrated the highest antimicrobial activity, with an inhibitory concentration of 32 µg/mL against C. jejuni and B. cereus. This study has shown that extracts of selected Australian plants possess antimicrobial activity against C. jejuni and thus may have application in the control of this organism in live poultry and retail poultry products.

  7. Ultrastructure of Campylobacter jejuni in gamma-irradiated mouse jejunum

    SciTech Connect

    Sosula, L.; Nicholls, E.M.; Skeen, M.

    1988-04-01

    This paper describes the ultrastructure of intracellular elongated, transitional and coccoid forms of Campylobacter jejuni, in irradiated mouse jejunum infected both in vitro and in vivo and in cultured human skin fibroblasts. Jejunum of irradiated mouse incubated for 1 hour under conditions favorable to the organisms showed minimal tissue degeneration. The intracellular organisms in this material were free cytoplasmic forms showing inner membrane degeneration, loss of cytoplasmic granules, and absence of flagella. The diameter of the coccoids was up to four times that of the elongated forms, as in plate cultures. Intracellular organisms were not found in challenged unirradiated controls, indicating that irradiation of mouse cells may be required for intracellular infection with human strains of C jejuni. In contrast, challenged human fibroblasts contained typical elongated organisms in cytoplasmic vacuoles. These findings are discussed with reference to Campylobacter strain, host resistance, and natural animal and human Campylobacter infections.

  8. Prevalence, virulence, and antimicrobial resistance of Campylobacter jejuni and Campylobacter coli in white stork Ciconia ciconia in Poland.

    PubMed

    Szczepańska, Bernadeta; Kamiński, Piotr; Andrzejewska, Małgorzata; Śpica, Dorota; Kartanas, Edmund; Ulrich, Werner; Jerzak, Leszek; Kasprzak, Mariusz; Bocheński, Marcin; Klawe, Jacek J

    2015-01-01

    The aim of this study was to investigate the role of white stork Ciconia ciconia as a potential reservoir of Campylobacter spp. Antimicrobial resistance and the presence of putative virulence genes of the isolates were also examined. A total of 398 white stork chicks sampled in Western Poland in habitats with high density of breeding were examined. Rectal swabs were collected during breeding season 2009-2012 from storks developing in a relatively pure environment (Odra meadows), in polluted areas (a copper mining-smelting complex), and in suburbs. Of the anal swabs collected, 7.6% were positive for Campylobacter among chicks (5.3% samples positive for C. jejuni and 2.3% samples positive for C. coli). Samples from polluted areas had the highest prevalence of Campylobacter (12.2%). The prevalence of resistance among C. jejuni and C. coli isolates from young storks was as follows: to ciprofloxacin (52.4%, 44.4%), and to tetracycline (19%, 77.8%). All of the analyzed isolates were susceptible to macrolides. The resistance to both classes of antibiotics was found in the 23.3% of Campylobacter spp. All Campylobacter spp. isolates had cadF gene and flaA gene responsible for adherence and motility. CdtB gene associated with toxin production was present in 88.9% of C. coli isolates and 57.1% of C. jejuni isolates. The iam marker was found more often in C. coli strains (55.6%) compared to C. jejuni isolates (42.9%). Our results confirm the prevalence of Campylobacter spp. in the white stork in natural conditions and, because it lives in open farmlands with access to marshy wetlands, the environmental sources such as water reservoirs and soil-water can be contaminated from white stork feces and the pathogens can be widely disseminated. We can thus conclude that Campylobacter spp. may easily be transmitted to waterfowl, other birds, and humans via its environmental sources and/or by immediate contact. PMID:25456607

  9. Prevalence, virulence, and antimicrobial resistance of Campylobacter jejuni and Campylobacter coli in white stork Ciconia ciconia in Poland.

    PubMed

    Szczepańska, Bernadeta; Kamiński, Piotr; Andrzejewska, Małgorzata; Śpica, Dorota; Kartanas, Edmund; Ulrich, Werner; Jerzak, Leszek; Kasprzak, Mariusz; Bocheński, Marcin; Klawe, Jacek J

    2015-01-01

    The aim of this study was to investigate the role of white stork Ciconia ciconia as a potential reservoir of Campylobacter spp. Antimicrobial resistance and the presence of putative virulence genes of the isolates were also examined. A total of 398 white stork chicks sampled in Western Poland in habitats with high density of breeding were examined. Rectal swabs were collected during breeding season 2009-2012 from storks developing in a relatively pure environment (Odra meadows), in polluted areas (a copper mining-smelting complex), and in suburbs. Of the anal swabs collected, 7.6% were positive for Campylobacter among chicks (5.3% samples positive for C. jejuni and 2.3% samples positive for C. coli). Samples from polluted areas had the highest prevalence of Campylobacter (12.2%). The prevalence of resistance among C. jejuni and C. coli isolates from young storks was as follows: to ciprofloxacin (52.4%, 44.4%), and to tetracycline (19%, 77.8%). All of the analyzed isolates were susceptible to macrolides. The resistance to both classes of antibiotics was found in the 23.3% of Campylobacter spp. All Campylobacter spp. isolates had cadF gene and flaA gene responsible for adherence and motility. CdtB gene associated with toxin production was present in 88.9% of C. coli isolates and 57.1% of C. jejuni isolates. The iam marker was found more often in C. coli strains (55.6%) compared to C. jejuni isolates (42.9%). Our results confirm the prevalence of Campylobacter spp. in the white stork in natural conditions and, because it lives in open farmlands with access to marshy wetlands, the environmental sources such as water reservoirs and soil-water can be contaminated from white stork feces and the pathogens can be widely disseminated. We can thus conclude that Campylobacter spp. may easily be transmitted to waterfowl, other birds, and humans via its environmental sources and/or by immediate contact.

  10. 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

  11. Strain characterization and grouping of Campylobacter jejuni and Campylobacter coli by interaction with lectins.

    PubMed Central

    Wong, K H; Skelton, S K; Feeley, J C

    1986-01-01

    Strains of Campylobacter jejuni and Campylobacter coli were characterized and grouped by their distinct reaction patterns with lectins. Heating of the Campylobacter cultures to 100 degrees C and holding for 30 to 60 min greatly enhanced their reactivity with lectins and permitted the grouping of all but 3 of 155 cultures tested in this study without interference of autoagglutination and other nonspecific activities. The lectin reaction patterns of the heated cultures were stable and reproducible. They were strain specific and independent of the heat-stable antigenic types. The lectin-reactive sites of C. jejuni and C. coli may be useful as additional markers for strain characterization. Based on these observations, a simple slide agglutination procedure is described for differentiating strains of C. jejuni and C. coli by their interaction with a selected group of commercially available lectins. PMID:3754264

  12. Human serum antibody response in Campylobacter jejuni enteritis as measured by enzyme-linked immunosorbent assay.

    PubMed

    Herbrink, P; van den Munckhof, H A; Bumkens, M; Lindeman, J; van Dijk, W C

    1988-06-01

    An ELISA for detection of IgG, IgA, and IgM antibody using an acid-glycine extract from Campylobacter jejuni as antigen was developed. To determine the value of this assay for the diagnosis of acute Campylobacter jejuni infections, the IgG, IgA, and IgM immune response against Campylobacter jejuni was investigated at various timepoints after infection in patients with culture-proven infection. A total of 112 sera from 46 patients and 78 sera from a control group were tested. All but one of the 46 patients with culture-proven Campylobacter jejuni enteritis developed IgG antibodies against Campylobacter jejuni. IgA and IgM ELISA both showed 97% specificity, and sensitivity of 63% and 30% respectively. IgG antibody titers generally remained at a constant level for more than 50 days, whereas IgA and IgM antibody titers declined more rapidly to normal values within 30 to 50 days after onset of clinical symptoms. Detection of Campylobacter jejuni specific IgA antibodies in a single serum sample provided the most useful assay for serological diagnosis of Campylobacter jejuni enteritis. The presence of Campylobacter jejuni specific IgM antibodies was the sole diagnostic criterion in three cases. Serological diagnosis of Campylobacter jejuni enteritis should therefore include both IgA and IgM antibody determination.

  13. Sensitive detection of Campylobacter jejuni using nanoparticles enhanced QCM sensor.

    PubMed

    Masdor, Noor Azlina; Altintas, Zeynep; Tothill, Ibtisam E

    2016-04-15

    A quartz crystal microbalance (QCM) sensor platform was used to develop an immunosensor for the detection of food pathogen Campylobacter jejuni. Rabbit polyclonal antibodies and commercially available mouse monoclonal antibodies against C. jejuni were investigated to construct direct, sandwich and gold-nanoparticles (AuNPs) amplified sandwich assays. The performance of the QCM immunosensor developed using sandwich assay by utilising the rabbit polyclonal antibody as the capture antibody and conjugated to AuNPs as the detection antibody gave the highest sensitivity. This sensor achieved a limit of detection (LOD) of 150 colony forming unit (CFU)mL(-1) of C. jejuni in solution. The QCM sensor showed excellent sensitivity and specificity for Campylobacter detection with low cross reactivity for other foodborne pathogens such as Salmonella Typhimurium, (7%) Listeria monocytogenes (3%) and Escherichia coli (0%). The development of this biosensor would help in the sensitive detection of Campylobacter which can result in reducing pre-enrichment steps; hence, reducing assay time. This work demonstrates the potential of this technology for the development of a rapid and sensitive detection method for C. jejuni.

  14. Sensitive detection of Campylobacter jejuni using nanoparticles enhanced QCM sensor.

    PubMed

    Masdor, Noor Azlina; Altintas, Zeynep; Tothill, Ibtisam E

    2016-04-15

    A quartz crystal microbalance (QCM) sensor platform was used to develop an immunosensor for the detection of food pathogen Campylobacter jejuni. Rabbit polyclonal antibodies and commercially available mouse monoclonal antibodies against C. jejuni were investigated to construct direct, sandwich and gold-nanoparticles (AuNPs) amplified sandwich assays. The performance of the QCM immunosensor developed using sandwich assay by utilising the rabbit polyclonal antibody as the capture antibody and conjugated to AuNPs as the detection antibody gave the highest sensitivity. This sensor achieved a limit of detection (LOD) of 150 colony forming unit (CFU)mL(-1) of C. jejuni in solution. The QCM sensor showed excellent sensitivity and specificity for Campylobacter detection with low cross reactivity for other foodborne pathogens such as Salmonella Typhimurium, (7%) Listeria monocytogenes (3%) and Escherichia coli (0%). The development of this biosensor would help in the sensitive detection of Campylobacter which can result in reducing pre-enrichment steps; hence, reducing assay time. This work demonstrates the potential of this technology for the development of a rapid and sensitive detection method for C. jejuni. PMID:26649490

  15. Higher resistance of Campylobacter coli compared to Campylobacter jejuni at chicken slaughterhouse.

    PubMed

    Torralbo, Alicia; Borge, Carmen; García-Bocanegra, Ignacio; Méric, Guillaume; Perea, Anselmo; Carbonero, Alfonso

    2015-04-01

    In order to compare the prevalence of Campylobacter coli and Campylobacter jejuni during the processing of broilers at slaughterhouse a total of 848 samples were analyzed during 2012 in southern Spain. Four hundred and seventy six samples were collected from cloaca, carcass surfaces and quartered carcasses. Moreover, 372 environmental swabs from equipment and scalding water were collected. Minimum inhibitory concentration (MIC) to ciprofloxacin, erythromycin, streptomycin, tetracycline and gentamicin was determined for isolates from chicken meat. The general prevalence of Campylobacter was 68.8% (40.2% of C. coli and 28.5% of C. jejuni). The relative prevalence of C. coli increased from loading dock area (41.5%) to packing area (64.6%). In contrast, the relative prevalence of C. jejuni decreased from 58.5% to 35.4%. These differences between species from initial to final area were significant (p=0.02). The highest antimicrobial resistance for C. jejuni and C. coli was detected to tetracycline (100%) and ciprofloxacin (100%), respectively. Campylobacter coli showed an antimicrobial resistance significantly higher than C. jejuni to streptomycin (p=0.002) and erythromycin (p<0.0001).

  16. Campylobacter jejuni Bacteremia in a Patient With Acute Lymphocytic Leukemia

    PubMed Central

    Anvarinejad, Mojtaba; Amin Shahidi, Maneli; Pouladfar, Gholam Reza; Dehyadegari, Mohammad Ali; Mardaneh, Jalal

    2016-01-01

    Introduction Campylobacter jejuni is a slender, motile, non-spore-forming, helical-shaped, gram-negative bacterium. It is one of the most common causes of human gastroenteritis in the world. The aim of this study was to present a patient with acute lymphocytic leukemia (ALL), who was infected with Campylobacter jejuni. Case Presentation We describe the medical records of a pediatric ALL patient with bacteremia caused by C. jejuni, who was diagnosed at Amir hospital, Shiraz, Iran. This 14-year-old male visited the emergency department of Amir hospital with night sweats, severe polar high-grade fever, reduced appetite, and nausea in August 2013. Given the suspected presence of an anaerobic or microaerophilic microorganism, aerobic and anaerobic blood cultures were performed using an automated blood cultivator, the BACTEC 9240 system. In order to characterize the isolate, diagnostic biochemical tests were used. Antibiotic susceptibility testing was done with the disk diffusion method. The primary culture was found to be positive for Campylobacter, and the subculture of the solid plate yielded a confluent growth of colonies typical for Campylobacter, which was identified as C. jejuni by morphological and biochemical tests. The isolate was resistant to ciprofloxacin, cefotaxime, cephalexin, piperacillin/tazobactam, nalidixic acid, aztreonam, cefuroxime, cefixime, ceftazidime, and tobramycin. Conclusions C. jejuni should be considered in the differential diagnosis as a potential cause of bacteremia in immunosuppressed patients. In cases where the BACTEC result is positive in aerobic conditions but the organism cannot be isolated, an anaerobic culture medium is suggested, especially in immunocompromised patients. PMID:27621914

  17. Stress response and virulence of heat-stressed Campylobacter jejuni.

    PubMed

    Klančnik, Anja; Vučković, Darinka; Jamnik, Polona; Abram, Maja; Možina, Sonja Smole

    2014-01-01

    Thermotolerant Campylobacter spp. frequently cause bacterial gastroenteritis in humans commonly infected through the consumption of undercooked poultry meat. We examined Campylobacter jejuni heat-stress responses in vitro after exposure to 48°C and 55°C. The in vivo modulation of its pathogenicity was also investigated using BALB/c mice intravenously infected with stressed C. jejuni. Regardless of the bacterial growth phase, the culturability and viability of C. jejuni in vitro was reduced after exposure to 55°C. This correlated with the altered protein profile and decreased virulence properties observed in vivo. Heat stress at 48°C elicited the transition to more resistant bacterial forms, independent of morphological changes or the appearance of shorter spiral and coccoid cells. This treatment did not cause marked changes in bacterial virulence properties in vivo. These results indicated that the characteristics and pathogenicity of C. jejuni in response to heat stress are temperature dependent. Further studies on the responses of C. jejuni to stresses used during food processing, as well as the modulation of its virulence, are important for a better understanding of its contamination and infective cycle, and will, thus, contribute to improved safety in the food production chain.

  18. Cellular Response of Campylobacter jejuni to Trisodium Phosphate

    PubMed Central

    Riedel, Charlotte Tandrup; Cohn, Marianne Thorup; Stabler, Richard A.; Wren, Brendan

    2012-01-01

    The highly alkaline compound trisodium phosphate (TSP) is used as an intervention to reduce the load of Campylobacter on poultry meat in U.S. poultry slaughter plants. The aim of the present study was to investigate the cellular responses of Campylobacter jejuni NCTC11168 when exposed to sublethal concentrations of TSP. Preexposure of C. jejuni to TSP resulted in a significant increase in heat sensitivity, suggesting that a combined heat and TSP treatment may increase reduction of C. jejuni. A microarray analysis identified a limited number of genes that were differently expressed after sublethal TSP exposure; however, the response was mainly associated with ion transport processes. C. jejuni NCTC11168 nhaA1 (Cj1655c) and nhaA2 (Cj1654c), which encode orthologues to the Escherichia coli NhaA cation/proton antiporter, were able to partially restore TSP, alkaline, and sodium resistance phenotypes to an E. coli cation/proton antiporter mutant. In addition, inhibition of resistance-nodulation-cell division (RND) multidrug efflux pumps by the inhibitor PaβN (Phe-Arg β-naphthylamide dihydrochloride) decreased tolerance to sublethal TSP. Therefore, we propose that NhaA1/NhaA2 cation/proton antiporters and RND multidrug efflux pumps function in tolerance to sublethal TSP exposure in C. jejuni. PMID:22194296

  19. Antimicrobial activity of two propolis samples against human Campylobacter jejuni.

    PubMed

    Campana, Raffaella; Patrone, Vania; Franzini, Ingrid Tarsilla Maria; Diamantini, Giuseppe; Vittoria, Emanuela; Baffone, Wally

    2009-10-01

    The aim of this study was to analyze the antimicrobial activity of two ethanolic extracts of propolis (EEPs) and selected flavonoids against 16 Campylobacter jejuni clinical isolates and several Gram-positive and Gram-negative human pathogens. The antimicrobial activity of EEPs and flavonoids was evaluated by the agar well diffusion method. The EEPs inhibited the growth of C. jejuni, Enterobacter faecalis, and Staphylococcus aureus. The most active flavonoid was galangin, with the highest percentage of sensitivity among C. jejuni strains (68.8%); lower percentages of sensitivity were observed for quercetin (50%). The minimal inhibitory concentrations (MICs) of EEPs and flavonoids for C. jejuni isolates were determined by the agar dilution method. EEPs showed MIC values of 0.3125-0.156 mg/mL for all C. jejuni strains; galangin and quercetin gave MICs ranging from 0.250 to 0.125 mg/mL. Thus propolis preparations could be used as support to traditional therapy for Campylobacter infection, especially when the antibiotic agents show no activity against this microorganism.

  20. Campylobacter jejuni in dairy cows and raw milk.

    PubMed Central

    Humphrey, T. J.; Beckett, P.

    1987-01-01

    Twelve herds of dairy cows were examined by rectal swabbing for the presence of Campylobacter jejuni. Ten herds were positive with the incidence of colonized animals ranging from 10 to 72% of those tested. With the exception of the two negative herds where mains water only was consumed, all animals drank from rivers or streams when grazing. There was no relationship between total and coliform counts and the presence of C. jejuni in raw milk. However, milk from one farm that consistently gave positive results had significantly higher Escherichia coli counts than other samples. PMID:3595744

  1. Whole-Genome Sequences of Agricultural, Host-Associated Campylobacter coli and Campylobacter jejuni Strains.

    PubMed

    Dutta, Vikrant; Altermann, Eric; Olson, Jonathan; Wray, Gregory Allan; Siletzky, Robin M; Kathariou, Sophia

    2016-01-01

    We report here the genome sequences of four agricultural, multidrug-resistant Campylobacter spp.: C. coli 11601 and C. jejuni 11601MD, isolated from turkey cecum and jejunum, respectively, and C. coli 6067 and C. coli 6461, isolated from turkey-house water and swine feces, respectively. The genomes provide insights on Campylobacter antimicrobial resistance and host adaptations. PMID:27540063

  2. Interaction of Campylobacter jejuni and Campylobacter coli with lectins and blood group antibodies.

    PubMed Central

    Wong, K H; Skelton, S K; Feeley, J C

    1985-01-01

    Lectins and blood group antibodies were used to probe the surface structures of Campylobacter jejuni and Campylobacter coli. Of the 29 strains tested, there were distinct reaction patterns. The lectin-reactive and blood group antibody-reactive sites on the bacterial surface were distinguishable from the heat-stable (lipopolysaccharide) antigenic determinants. The interactions were strain specific. The reactive sites were stable with respect to culture media and passage and may be useful as additional markers for strain characterization. PMID:2410445

  3. Whole-Genome Sequences of Agricultural, Host-Associated Campylobacter coli and Campylobacter jejuni Strains

    PubMed Central

    Altermann, Eric; Olson, Jonathan; Wray, Gregory Allan; Siletzky, Robin M.; Kathariou, Sophia

    2016-01-01

    We report here the genome sequences of four agricultural, multidrug-resistant Campylobacter spp.: C. coli 11601 and C. jejuni 11601MD, isolated from turkey cecum and jejunum, respectively, and C. coli 6067 and C. coli 6461, isolated from turkey-house water and swine feces, respectively. The genomes provide insights on Campylobacter antimicrobial resistance and host adaptations. PMID:27540063

  4. Isolation, characterization, and serotyping of Campylobacter jejuni and Campylobacter coli from slaughter cattle.

    PubMed Central

    Garcia, M M; Lior, H; Stewart, R B; Ruckerbauer, G M; Trudel, J R; Skljarevski, A

    1985-01-01

    A total of 525 specimens from 100 slaughter beef cattle were examined for the presence of Campylobacter jejuni and Campylobacter coli by direct plating and enrichment techniques. Isolates were identified by cultural, biochemical, antibiotic sensitivity, and immunofluorescence tests and further characterized with the aid of recently developed biotyping and serotyping methods. Fifty animals were positive for C. jejuni; only one was positive for C. coli. The distribution pattern of C. jejuni-positive animals, in decreasing order, was steers (55%), bulls (40%), heifers (40%), and cows (22%). Significantly higher isolation rates were obtained from the gall bladders (33%), large intestines (35%), and small intestines (31%) than from the livers (12%) or the lymph nodes (1.4%). C. jejuni isolation by the enrichment technique was 40.2% more frequent than by direct plating; 24-h enrichment resulted in 24% more isolations than 48-h enrichment. Eighty-four of 105 C. jejuni cultures were typable serologically and represented 13 serogroups. Biotype I accounted for 71% of biotyped cultures. Serogroup 7 biotype I was the most commonly encountered (24%) isolate. About one in three positive animals had C. jejuni strains representing more than one serogroup. C. jejuni serogroups encountered in slaughter cattle were similar to those commonly isolated from human sources. PMID:3888109

  5. Attenuation of Adhesion, Biofilm Formation and Quorum Sensing of Campylobacter jejuni by Euodia ruticarpa.

    PubMed

    Bezek, Katja; Kurinčič, Marija; Knauder, Elvira; Klančnik, Anja; Raspor, Peter; Bucar, Franz; Smole Možina, Sonja

    2016-09-01

    Thermophilic campylobacters are a major cause of bacterial food-borne diarrhoeal disease. Adherence and biofilm formation are key elements of Campylobacter jejuni persistence in unfavourable environmental conditions. The phytochemical analysis of Euodia ruticarpa fruit ethanol solution extract (EREE) indicated that the major compounds were evodiamine (1), rutaecarpine (2) and evocarpine (9). E. ruticarpa fruit ethanol solution extract, compounds 1 and 2 as well as a mixture of quinolinone alkaloids with 41.7% of 9 were tested for antibacterial, antibiofilm and antiquorum sensing activities against C. jejuni. Minimal inhibitory concentrations varied from 64 to 1024 µg/mL. A mutant strain that lacks the functional gene coding for the CmeB efflux pump protein was the most susceptible. Interestingly, in addition to the wild-type (NCTC 11168) and cmeB mutant, also a mutant that lacks autoinducer-2 production (luxS) was able to adhere (1 h) and to produce a biofilm (24, 48 and 72 h). The subinhibitory concentrations of all preparations at least partly inhibited C. jejuni adhesion and biofilm formation with the most visible effect of the quinolinone alkaloid fraction. Using a Vibrio harveyi luminescence assay, the inhibition of autoinducer-2 production was observed in the wild-type and cmeB mutant after 48 h with the most visible effect of EREE and its fraction Q. Copyright © 2016 John Wiley & Sons, Ltd. PMID:27230628

  6. The immunobiology of Campylobacter jejuni: Innate immunity and autoimmune diseases.

    PubMed

    Phongsisay, Vongsavanh

    2016-04-01

    The Gram-negative bacterium Campylobacter jejuni causes gastroenteritis and Guillain-Barré syndrome in humans. Recent advances in the immunobiology of C. jejuni have been made. This review summarizes C. jejuni-binding innate receptors and highlights the role of innate immunity in autoimmune diseases. This human pathogen produces a variety of glycoconjugates, including human ganglioside-like determinants and multiple activators of Toll-like receptors (TLRs). Furthermore, C. jejuni targets MyD88, NLRP3 inflammasome, TIR-domain-containing adapter-inducing interferon-β (TRIF), sialic acid-binding immunoglobulin-like lectins (Siglecs), macrophage galactose-type lectin (MGL), and immunoglobulin-like receptors (TREM2, LMIR5/CD300b). The roles of these innate receptors and signaling molecules have been extensively studied. MyD88-mediated TLR activation or inflammasome-dependent IL-1β secretion is essential for autoimmune induction. TRIF mediates the production of type I interferons that promote humoral immune responses and immunoglobulin class-switching. Siglec-1 and Siglec-7 interact directly with gangliosides. Siglec-1 activation enhances phagocytosis and inflammatory responses. MGL internalizes GalNAc-containing glycoconjugates. TREM2 is well-known for its role in phagocytosis. LMIR5 recognizes C. jejuni components and endogenous sulfoglycolipids. Several lines of evidence from animal models of autoimmune diseases suggest that simultaneous activation of innate immunity in the presence of autoreactive lymphocytes or antigen mimicry may link C. jejuni to immunopathology.

  7. [Two cases of enteritis with bacteremia due to Campylobacter jejuni].

    PubMed

    Borda, Noemí; Gambandé, Telma; Notario, Rodolfo

    2006-01-01

    Campylobacter is an important agent of illness in human beings. Bacteremia occurs principally in the immunocompromissed host and is frequently due to C. fetus. Nevertheless bacteremia also has been observed in patients with enteritis due to C. jejuni. We refer two cases of patients with severe enteritis and bacteremia, both of them with immunosupressive concomitant diseases such as nephrotic syndrome and chronic cirrotic hepatopathy. Both patients presented hemathemesis.

  8. Susceptibility of Campylobacter jejuni and Yersinia enterocolitica to UV radiation

    SciTech Connect

    Butler, R.C.; Lund, V.; Carlson, D.A.

    1987-02-01

    Two enteric pathogens, Campylobacter jejuni and Yersinia enterocolitica serogroup O:3, together with Escherichia coli, were investigated for susceptibility to UV radiation at 254 nm. The UV dose required for a 3-log reduction (99.9% inactivation) of C. jejuni, Y. enterocolitica, and E. coli was 1.8, 2.7, and 5.0 mWs/cm2, respectively. Using E. coli as the basis for comparison, it appears that C. jejuni and Y. enterocolitica serogroup O:3 are more sensitive to UV than many of the pathogens associated with waterborne disease outbreaks and can be easily inactivated in most commercially available UV reactors. No association was found between the sensitivity of Y. enterocolitica to UV and the presence of a 40- to 50-megadalton virulence plasmid.

  9. Susceptibility of Campylobacter jejuni and Yersinia enterocolitica to UV radiation.

    PubMed Central

    Butler, R C; Lund, V; Carlson, D A

    1987-01-01

    Two enteric pathogens, Campylobacter jejuni and Yersinia enterocolitica serogroup O:3, together with Escherichia coli, were investigated for susceptibility to UV radiation at 254 nm. The UV dose required for a 3-log reduction (99.9% inactivation) of C. jejuni, Y. enterocolitica, and E. coli was 1.8, 2.7, and 5.0 mWs/cm2, respectively. Using E. coli as the basis for comparison, it appears that C. jejuni and Y. enterocolitica serogroup O:3 are more sensitive to UV than many of the pathogens associated with waterborne disease outbreaks and can be easily inactivated in most commercially available UV reactors. No association was found between the sensitivity of Y. enterocolitica to UV and the presence of a 40- to 50-megadalton virulence plasmid. PMID:3551844

  10. Immunogens of interest for the diagnosis of Campylobacter jejuni infections.

    PubMed

    Janvier, B; Ayraud, S; Beby-Defaux, A; Louis Fauchère, J

    2000-03-01

    In order to identify the C. jejuni immunogens of interest for the diagnosis of Campylobacter infections, we analyzed the humoral response of 153 patients by using complement fixation (CF) and western blot assays. A first group of 79 sera was from C. jejuni infected patients suffering from enteritis (n=16), Guillain-Barré syndrome (GBS) (n=40) and arthritis (n=23). A second group of 49 sera was from healthy blood donors and a third group consisted of 25 sera from children under 4 years old. Using the CF test, 88.6% of the C. jejuni infected patients were seropositive versus 28.5% of the healthy blood donors and none of the children. The Western blot assay allowed detection of antibodies directed against seven selected antigens ranging from 14 to 67 kDa. Three of these antigens with a molecular size of 29, 37 and 43 kDa were detected by 86.0%, 84.8% and 91.1% of the C. jejuni infected patients, respectively. These three antigens seem to be good candidates for the development of assays suitable for direct and indirect diagnosis of Campylobacter infections.

  11. Biotypes, serogroups and antibiotic susceptibility of Campylobacter jejuni and Campylobacter coli in Chile.

    PubMed

    Figueroa, G; Troncoso, M; Galeno, H; Soto, V; Toledo, M S

    1990-03-01

    Phenotypic markers were studied in 105 strains of thermophilic campylobacters isolated from human beings, animals and drinking water in Santiago, Chile. The strains were identified as Campylobacter jejuni (n = 49) and Campylobacter coli (n = 56). Biotypes I and II (Lior schema) accounted for 96% C. jejuni isolates, the other 4% being biotype IV but the two biotypes of C. coli were about equally represented. A total of 28 serogroups (Lior's heat-labile antigens) were identified. Lior 13, 9, 79, 2 and 4 were prevalent among the C. jejuni, while Lior 8, 21 and 29/75 were prevalent among the C. coli isolates. These serogroups accounted for 73% all isolates. The distribution of biotypes and serogroups in patients and asymptomatic persons were similar. Human campylobacters were often resistant to ampicillin (31%) but sensitive to erythromycin and furazolidone. Swine C. coli isolates proved resistant to streptomycin (46%), tetracycline (38%) and erythromycin (15%). Determination of phenotypic and serological characters provides valuable epidemiological markers in the study of campylobacter infections.

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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, ...

  13. Prevalence of virulence genes in strains of Campylobacter jejuni isolated from human, bovine and broiler.

    PubMed

    González-Hein, Gisela; Huaracán, Bernardo; García, Patricia; Figueroa, Guillermo

    2013-12-01

    Campylobacter jejuni isolates of different origins (bovine, broiler meat, human) were screened by polymerase chain reaction for the presence of 4 genes cdtB, cst-II, ggt, and virB11, previously linked to virulence such as adherence, invasion, colonization, molecular mimicry, and cytotoxin production. In addition, the isolates were screened for the presence of the global gene regulator csrA linked to oxidative stress responses, biofilms formation, and cell adhesion. All the C. jejuni isolates were positive for cdtB gene. The csrA gene was detected in 100% and 92% of C. jejuni isolates from human and animal origin and the virB11 gene was detected in 7.3% and 3.6% isolates from chicken and human respectively. All isolates from bovine were negative for the virB11 gene. The isolates showed a wide variation for the presence of the remaining genes. Of the C. jejuni recovered from human 83.6%, and 32.7% were positive for cst-II, and ggt respectively. Out of the isolates from chicken 40% and 5.5% isolates revealed the presence of cst-II, and ggt, respectively. Finally of the C. jejuni isolates from bovine, 97.7% and 22.7% were positive for cst-II, and ggt respectively. We conclude that the genes of this study circulate among humans and animals. These results led us to hypothesize that the isolates associated with enteritis (cdtB positives) are not selected by environmental or host-specific factors. On the other hand, the high frequencies of csrA gene in C. jejuni show that this gene is important for the survival of C. jejuni in animals and humans.

  14. Experimental colonization of broiler chicks with Campylobacter jejuni.

    PubMed Central

    Shanker, S.; Lee, A.; Sorrell, T. C.

    1988-01-01

    Minimal colonization inocula for two broiler strains of Campylobacter jejuni were determined in broiler chicks aged 2-3 days and 2 weeks. Individually housed chicks were exposed to a single oral or cloacal challenge. Diarrhoeal symptoms were absent in all 380 chicks included in the study. Chick susceptibility to the two C. jejuni strains varied. Colonization was effected by less than 10(2)-10(4) colony forming units (c.f.u.) via cloacal challenge and 10(4)-10(6) c.f.u. via the oral route. Colonization inocula for 2- to 3-day and 2-week-old chicks were similar. Treatment of 1-day-old chicks with fresh adult caecal flora or an anaerobic broth culture of adult caecal flora did not inhibit colonization after challenge with low-dose C. jejuni. Susceptible chicks were colonized rapidly. C. jejuni was detected in 167 of 189 (88%) colonized chicks within 3 days of challenge and persisted during the 2-week monitoring period. Our data suggest that colonization of broiler chicks with C. jejuni is effected more easily by the cloacal than the oral route and is independent of age. PMID:3338504

  15. Antimicrobial effect of diallyl sulphide on Campylobacter jejuni biofilms

    PubMed Central

    Lu, Xiaonan; Samuelson, Derrick R.; Rasco, Barbara A.; Konkel, Michael E.

    2012-01-01

    Objectives Bacterial biofilms pose significant food safety risks because of their attachment to fomites and food surfaces, including fresh produce surfaces. The purpose of this study was to systematically investigate the activity of selected antimicrobials on Campylobacter jejuni biofilms. Methods C. jejuni biofilms and planktonic cells were treated with ciprofloxacin, erythromycin and diallyl sulphide and examined using infrared and Raman spectroscopies coupled with imaging analysis. Results Diallyl sulphide eliminated planktonic cells and sessile cells in biofilms at a concentration that was at least 100-fold less than used for either ciprofloxacin or erythromycin on the basis of molarity. Distinct cell lysis was observed in diallyl sulphide-treated planktonic cells using immunoblot analysis and was confirmed by a rapid decrease in cellular ATP. Two phases of C. jejuni biofilm recalcitrance modes against ciprofloxacin and erythromycin were validated using vibrational spectroscopies: (i) an initial hindered adsorption into biofilm extracellular polymeric substance (EPS) and delivery of antibiotics to sessile cells within biofilms; and (ii) a different interaction between sessile cells in a biofilm compared with their planktonic counterparts. Diallyl sulphide destroyed the EPS structure of the C. jejuni biofilm, after which the sessile cells were killed in a similar manner as planktonic cells. Spectroscopic models can predict the survival of sessile cells within biofilms. Conclusions Diallyl sulphide elicits strong antimicrobial activity against planktonic and sessile C. jejuni and may have applications for reducing the prevalence of this microbe in foods, biofilm reduction and, potentially, as an alternative chemotherapeutic agent for multidrug-resistant bacterial strains. PMID:22550133

  16. Bacteriocins control chicken colonization while probiotic bacteria are ineffective at reducing Campylobacter jejuni

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Broilers chickens are widely considered an important source for human exposure to Campylobacter jejuni. We sought to intervene in C. jejuni colonization by using a probiotic approach. Isolates from chicken intestine were screened for C. jejuni inhibition. These isolates were live-fed to treat chi...

  17. The Campylobacter jejuni RacRS system regulates fumarate utilization in a low oxygen environment

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The natural environment of the human pathogen Campylobacter jejuni is the gastrointestinal tract of warm blooded animals. In the gut, the availability of oxygen is limited; therefore, less efficient electron acceptors such as nitrate or fumarate are used by C. jejuni. C. jejuni has a highly branched...

  18. Structural Heterogeneity of Terminal Glycans in Campylobacter jejuni Lipooligosaccharides

    PubMed Central

    Semchenko, Evgeny A.; Day, Christopher J.; Moutin, Marc; Wilson, Jennifer C.; Tiralongo, Joe; Korolik, Victoria

    2012-01-01

    Lipooligosaccharides of the gastrointestinal pathogen Campylobacter jejuni are regarded as a major virulence factor and are implicated in the production of cross-reactive antibodies against host gangliosides, which leads to the development of autoimmune neuropathies such as Guillain-Barré and Fisher Syndromes. C. jejuni strains are known to produce diverse LOS structures encoded by more than 19 types of LOS biosynthesis clusters. This study demonstrates that the final C. jejuni LOS structure cannot always be predicted from the genetic composition of the LOS biosynthesis cluster, as determined by novel lectin array analysis of the terminal LOS glycans. The differences were shown to be partially facilitated by the differential on/off status of three genes wlaN, cst and cj1144-45. The on/off status of these genes was also analysed in C. jejuni strains grown in vitro and in vivo, isolated directly from the host animal without passaging, using immunoseparation. Importantly, C. jejuni strains 331, 421 and 520 encoding cluster type C were shown to produce different LOS, mimicking asialo GM1, asialo GM2 and a heterogeneous mix of gangliosides and other glycoconjugates respectively. In addition, individual C. jejuni colonies were shown to consistently produce heterogeneous LOS structures, irrespective of the cluster type and the status of phase variable genes. Furthermore we describe C. jejuni strains (351 and 375) with LOS clusters that do not match any of the previously described LOS clusters, yet are able to produce LOS with asialo GM2-like mimicries. The LOS biosynthesis clusters of these strains are likely to contain genes that code for LOS biosynthesis machinery previously not identified, yet capable of synthesising LOS mimicking gangliosides. PMID:22815868

  19. Structural heterogeneity of terminal glycans in Campylobacter jejuni lipooligosaccharides.

    PubMed

    Semchenko, Evgeny A; Day, Christopher J; Moutin, Marc; Wilson, Jennifer C; Tiralongo, Joe; Korolik, Victoria

    2012-01-01

    Lipooligosaccharides of the gastrointestinal pathogen Campylobacter jejuni are regarded as a major virulence factor and are implicated in the production of cross-reactive antibodies against host gangliosides, which leads to the development of autoimmune neuropathies such as Guillain-Barré and Fisher Syndromes. C. jejuni strains are known to produce diverse LOS structures encoded by more than 19 types of LOS biosynthesis clusters. This study demonstrates that the final C. jejuni LOS structure cannot always be predicted from the genetic composition of the LOS biosynthesis cluster, as determined by novel lectin array analysis of the terminal LOS glycans. The differences were shown to be partially facilitated by the differential on/off status of three genes wlaN, cst and cj1144-45. The on/off status of these genes was also analysed in C. jejuni strains grown in vitro and in vivo, isolated directly from the host animal without passaging, using immunoseparation. Importantly, C. jejuni strains 331, 421 and 520 encoding cluster type C were shown to produce different LOS, mimicking asialo GM(1), asialo GM(2) and a heterogeneous mix of gangliosides and other glycoconjugates respectively. In addition, individual C. jejuni colonies were shown to consistently produce heterogeneous LOS structures, irrespective of the cluster type and the status of phase variable genes. Furthermore we describe C. jejuni strains (351 and 375) with LOS clusters that do not match any of the previously described LOS clusters, yet are able to produce LOS with asialo GM(2)-like mimicries. The LOS biosynthesis clusters of these strains are likely to contain genes that code for LOS biosynthesis machinery previously not identified, yet capable of synthesising LOS mimicking gangliosides. PMID:22815868

  20. High Prevalence and Genetic Diversity of Campylobacter jejuni in Wild Crows and Pigeons.

    PubMed

    Ramonaitė, Sigita; Novoslavskij, Aleksandr; Zakarienė, Gintarė; Aksomaitienė, Jurgita; Malakauskas, Mindaugas

    2015-11-01

    The occurrence, seasonal variation and genetic diversity of Campylobacter spp. in pigeons and crows over a 1-year period were evaluated. Campylobacter spp. were isolated from 166 (34.6 %) out of 480 wild bird faecal samples. The occurrence of Campylobacter spp. in faecal samples was higher among crows (39.2 %) than pigeons (30.0 %), (P < 0.05). Campylobacter jejuni was the most common species detected among wild bird faecal samples (98.2 %). Meanwhile, Campylobacter coli prevalence in wild bird faecal samples was low-6 %. The Simpson's diversity index of C. jejuni flaA RFLP types was lower in pigeons (D = 0.88) compared with C. jejuni isolates detected in crows (D = 0.97). Obtained results revealed that C. jejuni are widely prevalent among crows and pigeons, indicating these wild birds as potential infection sources to humans. Further studies are required to determine crows and pigeons role in zoonotic transmission of Campylobacter.

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

    PubMed

    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.

  2. Adhesion of Campylobacter jejuni and Mycobacterium avium onto polyethylene terephtalate (PET) used for bottled waters.

    PubMed

    Tatchou-Nyamsi-König, Josiane-Aurore; Dague, Etienne; Mullet, Martine; Duval, Jérôme F L; Gaboriaud, Fabien; Block, Jean-Claude

    2008-12-01

    Adhesion of the bacteria Campylobacter jejuni and Mycobacterium avium onto polyethylene terephtalate (PET), a polymer widely used within the bottled water industry was measured in two different groundwater solutions. From this, it was found that whilst the percentage cell adhesion for a given strain did not change between groundwater types, substantial variation was obtained between the two bacterial species tested: M. avium (10-30% adhered cells) and C. jejuni (1-2%) and no major variations were measured as a function of groundwater composition for a given strain. To explain this, the interfacial electro-hydrodynamic properties of the bacteria were investigated by microelectrophoresis, with the resultant data analysed on the basis of electrokinetic theory for soft biocolloidal particles. The results obtained showed that M. avium carries a significant volume charge density and that its peripheral layer exhibits limited hydrodynamic flow permeation compared to that of C. jejuni. It was also demonstrated that steric hindrance to flow penetration and the degree of hydrophobicity within/of the outer bacterial interface are larger for M. avium cells. In line with this, the larger amount of M. avium cells deposited onto PET substrates as compared to that of C. jejuni can be explained by hydrophobic attraction and chemical binding between hydrophobic PET and outer soft surface layer of the bacteria. Hydrophobicity of PET was addressed by combining contact angle analyses and force spectroscopy using CH(3)-terminated AFM tip. PMID:18929388

  3. Bacteriophage receptor binding protein based assays for the simultaneous detection of Campylobacter jejuni and Campylobacter coli.

    PubMed

    Javed, Muhammad A; Poshtiban, Somayyeh; Arutyunov, Denis; Evoy, Stephane; Szymanski, Christine M

    2013-01-01

    Campylobacter jejuni and Campylobacter coli are the most common bacterial causes of foodborne gastroenteritis which is occasionally followed by a debilitating neuropathy known as Guillain-Barré syndrome. Rapid and specific detection of these pathogens is very important for effective control and quick treatment of infection. Most of the diagnostics available for these organisms are time consuming and require technical expertise with expensive instruments and reagents to perform. Bacteriophages bind to their host specifically through their receptor binding proteins (RBPs), which can be exploited for pathogen detection. We recently sequenced the genome of C. jejuni phage NCTC12673 and identified its putative host receptor binding protein, Gp047. In the current study, we localized the receptor binding domain to the C-terminal quarter of Gp047. CC-Gp047 could be produced recombinantly and was capable of agglutinating both C. jejuni and C. coli cells unlike the host range of the parent phage which is limited to a subset of C. jejuni isolates. The agglutination procedure could be performed within minutes on a glass slide at room temperature and was not hindered by the presence of buffers or nutrient media. This agglutination assay showed 100% specificity and the sensitivity was 95% for C. jejuni (n = 40) and 90% for C. coli (n = 19). CC-Gp047 was also expressed as a fusion with enhanced green fluorescent protein (EGFP). Chimeric EGFP_CC-Gp047 was able to specifically label C. jejuni and C. coli cells in mixed cultures allowing for the detection of these pathogens by fluorescent microscopy. This study describes a simple and rapid method for the detection of C. jejuni and C. coli using engineered phage RBPs and offers a promising new diagnostics platform for healthcare and surveillance laboratories.

  4. Antimicrobial susceptibilities of Campylobacter jejuni and Campylobacter coli recovered from organic turkey farms in Germany.

    PubMed

    El-Adawy, Hosny; Ahmed, Marwa F E; Hotzel, Helmut; Tomaso, Herbert; Tenhagen, Bernd-Alois; Hartung, Joerg; Neubauer, Heinrich; Hafez, Hafez M

    2015-11-01

    The popularity of food produced from animals kept under an organic regimen has increased in recent years. In Germany, turkey meat consumption has increased. Despite several studies assessing the susceptibility of campylobacters to various antibiotics in poultry, no sufficient data exists regarding the antimicrobial resistance of campylobacters in organic-reared turkeys. This study provides information about antibiotic resistance in Campylobacter isolated from turkeys reared on organic farms in Germany. Ninety-six Campylobacter strains (41 C. jejuni and 55 C. coli) were isolated from different free-range turkey flocks. In vitro antimicrobial sensitivity testing was done using a broth microdilution test, and the presence of resistance genes to antibiotics (ciprofloxacin, tetracycline) was investigated. All Campylobacter isolates from organic turkeys (n = 96) were phenotypically sensitive to gentamicin, erythromycin, streptomycin, and chloramphenicol. In this study, the antibiotic susceptibilities of C. jejuni to ciprofloxacin, tetracycline, and naladixic acid were 56.0%, 51.3%, and 56.0%, respectively. In contrast, 44.0%, 73.0%, and 74.6% of C. coli isolates were resistant to tetracycline, ciprofloxacin, and nalidixic acid, respectively. Replacement of the Thr-86→Ile in the gyrA gene, and the presence of the tet(O) gene were the mainly identified resistance mechanisms against fluoroquinolones and tetracycline, respectively.These results also reinforce the need to develop strategies and implement specific control procedures to reduce the development of antimicrobial resistance.

  5. Growth and survival characteristics of Campylobacter jejuni in liquid egg.

    PubMed Central

    Hänninen, M. L.; Korkeala, H.; Pakkala, P.

    1984-01-01

    Growth and survival of four Campylobacter jejuni strains in yolk, in liquid whole egg and in white during aerobic storage at 37, 20 and 4 degrees C was followed. In 48 h at 37 degrees C the cell counts of C. jejuni increased by about 3 log10 units in yolk and 1.60-3.35(10) log units in liquid whole egg. The growth of C. jejuni was slightly better in yolk than in liquid whole egg. At 20 degrees C during 48 h the cell counts decreased by about 0.5-1.5 log10 units in yolk and in liquid whole egg. At 4 degrees C the decrease in cell counts after 21 days ranged from 1 to 2 log10 units, except for one strain, KH3, which could not be detected after 14 days storage in yolk. In liquid whole egg the cell counts of this strain also decreased considerably during storage. In white the number of inoculated C. jejuni cells decreased rapidly. The killing effect of white was shown to be temperature-dependent; at 37 and 20 degrees C no positive samples were detected after 24 h and at 4 degrees C no positive samples were found after 48 h. PMID:6537959

  6. 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

  7. Cryptic ecology among host generalist Campylobacter jejuni in domestic animals.

    PubMed

    Sheppard, Samuel K; Cheng, Lu; Méric, Guillaume; de Haan, Caroline P A; Llarena, Ann-Katrin; Marttinen, Pekka; Vidal, Ana; Ridley, Anne; Clifton-Hadley, Felicity; Connor, Thomas R; Strachan, Norval J C; Forbes, Ken; Colles, Frances M; Jolley, Keith A; Bentley, Stephen D; Maiden, Martin C J; Hänninen, Marja-Liisa; Parkhill, Julian; Hanage, William P; Corander, Jukka

    2014-05-01

    Homologous recombination between bacterial strains is theoretically capable of preventing the separation of daughter clusters, and producing cohesive clouds of genotypes in sequence space. However, numerous barriers to recombination are known. Barriers may be essential such as adaptive incompatibility, or ecological, which is associated with the opportunities for recombination in the natural habitat. Campylobacter jejuni is a gut colonizer of numerous animal species and a major human enteric pathogen. We demonstrate that the two major generalist lineages of C. jejuni do not show evidence of recombination with each other in nature, despite having a high degree of host niche overlap and recombining extensively with specialist lineages. However, transformation experiments show that the generalist lineages readily recombine with one another in vitro. This suggests ecological rather than essential barriers to recombination, caused by a cryptic niche structure within the hosts.

  8. Reduction of Campylobacter jejuni and Campylobacter coli in poultry skin by fruit extracts.

    PubMed

    Valtierra-Rodríguez, Diana; Heredia, Norma L; García, Santos; Sánchez, Eduardo

    2010-03-01

    Campylobacter spp. are a major cause of foodborne bacterial gastroenteritis in humans, and current methods to control Campylobacter contamination in foods are not completely successful. Plants are a promising source of antimicrobial agents, particularly given the growing interest in "all natural" foods. In this study, the antimicrobial activity of extracts from 28 edible plants against Campylobacter jejuni and Campylobacter coli was evaluated in vitro and in a poultry skin model. Nine of 28 extracts exhibited antimicrobial activity in a diffusion assay, and MBCs were determined for the three most active extracts, i.e., lime, plum, and sour orange peel (MBCs of 2 to 3 mg/ml). Mixtures of the lime, plum, and sour orange peel extracts were applied to chicken skin inoculated with 10(5) CFU of Campylobacter to test for synergistic or antagonist effects. After incubation (48 h at 4 degrees C) with any extract mixture, no Campylobacter CFUs were detectable. A panel of tasters determined that the mixture of lime and plum gave the best flavor to chicken wings. These active extracts from edible fruits are simple to prepare and are alternatives to reduce or eliminate Campylobacter contamination of chicken products. PMID:20202332

  9. Investigations into the antiadhesive activity of herbal extracts against Campylobacter jejuni.

    PubMed

    Bensch, K; Tiralongo, J; Schmidt, K; Matthias, A; Bone, K M; Lehmann, R; Tiralongo, E

    2011-08-01

    Campylobacter jejuni is one of the most common bacterial causes of diarrhoea in the industrialized world, being associated with the occurrence of Guillain-Barré Syndrome, and inducing diseases partially through intestinal adherence. With increasing reports of C. jejuni drug resistance against standard antibiotics, investigations into antiadhesive agents for the prevention of bacterial infection are highly significant. Given the consumer-driven development towards holistic and integrative healthcare, research into additional anti-Campylobacter effects of herbal medicines that are already used for their beneficial effects on bowel and digestive functions is important. Twenty-one herbal extracts were screened for antiadhesive activity against C. jejuni using modifications of previously published antiadhesion assays. Antiadhesion effects with IC(50) values <3 mg/mL were obtained for seven ethanol plant extracts, with Zingiber officinale (ginger), Capsicum annum (cayenne) and Glycyrrhiza glabra (licorice) displaying the highest antiadhesion activity against C. jejuni (IC(50) : <0.1 mg/mL, 0.29 mg/mL and 0.65 mg/mL, respectively). Differences in antiadhesion activity were found for two different Echinacea species, with E. purpurea displaying significantly higher and dose dependent antiadhesion activity than E. angustifolia. No significant antiadhesion activity (IC(50) values >35 mg/mL) was found for Agrimonia eupatoria (agrimony), Andrographis paniculata (andrographis), Matricaria recutita (chamomile), Foeniculum vulgare (fennel), Filipendula ulmaria (meadowsweet) and Artemisia absinthium (wormwood) extracts. This study provides evidence for additional beneficial effects of marketed herbal medicines in gastrointestinal disorders. PMID:21280113

  10. Relationship between Presence of Anti-Campylobacter FliD Protein Antibodies and Campylobacter jejuni Isolation from Broiler Chickens

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Campylobacter jejuni, a Gram-negative rod, is a zoonotic pathogen associated with human acute bacterial gastroenteritis worldwide. Poultry products are regarded as a major source of this bacterium for human infection. Although this bacterium is a commensal in chicken cecal microbiome, Campylobacte...

  11. Prevalence of Campylobacter jejuni, Campylobacter lari, and Campylobacter coli in Different Ecological Guilds and Taxa of Migrating Birds†

    PubMed Central

    Waldenström, Jonas; Broman, Tina; Carlsson, Inger; Hasselquist, Dennis; Achterberg, René P.; Wagenaar, Jaap A.; Olsen, Björn

    2002-01-01

    A total of 1,794 migrating birds trapped at a coastal site in southern Sweden were sampled for detection of Campylobacter spp. All isolates phenotypically identified as Campylobacter jejuni and a subset of those identified as non-C. jejuni were identified to the species level by PCR-based techniques. C. jejuni was found in 5.0% of the birds, Campylobacter lari was found in 5.6%, and Campylobacter coli was found in 0.9%. An additional 10.7% of the tested birds were infected with hippurate hydrolysis-negative Campylobacter spp. that were not identified to the species level. The prevalence of Campylobacter spp. differed significantly between ecological guilds of birds. Shoreline-foraging birds feeding on invertebrates and opportunistic feeders were most commonly infected (76.8 and 50.0%, respectively). High prevalence was also shown in other ground-foraging guilds, i.e., ground-foraging invertebrate feeders (11.0%), ground-foraging insectivores (20.3%), and plant-eating species (18.8%). Almost no Campylobacter spp. were found in ground-foraging granivores (2.3%), arboreal insectivores (0.6%), aerial insectivores (0%), or reed- and herbaceous plant-foraging insectivores (3.5%). During the autumn migration, a high proportion of samples from juveniles were positive (7.1% in passerines, 55.0% in shorebirds), indicating transmission on the breeding grounds or during the early part of migration. Prevalence of Campylobacter spp. was associated with increasing body mass among passerine bird species. Furthermore, prevalence was higher in short-distance migrants wintering in Europe than in long-distance migrants wintering in Africa, the Middle East, or Asia. Among ground-foraging birds of the Muscicapidae, those of the subfamily Turdinae (i.e., Turdus spp.) showed a high prevalence of Campylobacter spp., while the organism was not isolated in any member of the subfamily Muscicapinae (i.e., Erithacus and Luscinia). The prevalence of Campylobacter infection in wild birds thus

  12. Quantifying potential sources of surface water contamination with Campylobacter jejuni and Campylobacter coli.

    PubMed

    Mughini-Gras, Lapo; Penny, Christian; Ragimbeau, Catherine; Schets, Franciska M; Blaak, Hetty; Duim, Birgitta; Wagenaar, Jaap A; de Boer, Albert; Cauchie, Henry-Michel; Mossong, Joel; van Pelt, Wilfrid

    2016-09-15

    Campylobacter is the most common causative agent of human bacterial gastroenteritis and is frequently found in surface water, where it indicates recent contamination with animal faeces, sewage effluent, and agricultural run-off. The contribution of different animal reservoirs to surface water contamination with Campylobacter is largely unknown. In the Netherlands, the massive poultry culling to control the 2003 avian influenza epidemic coincided with a 44-50% reduction in human campylobacteriosis cases in the culling areas, suggesting substantial environment-mediated spread of poultry-borne Campylobacter. We inferred the origin of surface water Campylobacter jejuni and Campylobacter coli strains in Luxembourg and the Netherlands, as defined by multilocus sequence typing, by comparison to strains from poultry, pigs, ruminants, and wild birds, using the asymmetric island model for source attribution. Most Luxembourgish water strains were attributed to wild birds (61.0%), followed by poultry (18.8%), ruminants (15.9%), and pigs (4.3%); whereas the Dutch water strains were mainly attributed to poultry (51.7%), wild birds (37.3%), ruminants (9.8%), and pigs (1.2%). Attributions varied over seasons and surface water types, and geographical variation in the relative contribution of poultry correlated with the magnitude of poultry production at either the national or provincial level, suggesting that environmental dissemination of Campylobacter from poultry farms and slaughterhouses can be substantial in poultry-rich regions. PMID:27244295

  13. Serotyping scheme for Campylobacter jejuni and Campylobacter coli based on direct agglutination of heat-stable antigens.

    PubMed

    Frost, J A; Oza, A N; Thwaites, R T; Rowe, B

    1998-02-01

    Campylobacter is now the most frequently reported cause of gastrointestinal disease in England and Wales, yet few isolates are characterized beyond the genus level. The majority of isolates are Campylobacter jejuni (90%), with most of the remainder being Campylobacter coli. We describe an adaptation of the Penner serotyping scheme in which passive hemagglutination has been replaced by detection of heat-stable antigens by direct bacterial agglutination; absorbed antisera are used where appropriate. This scheme has been used to type 2,407 C. jejuni samples and 182 C. coli samples isolated in Wales between April 1996 and March 1997. Forty-seven C. jejuni serotypes were identified, with the 10 most prevalent serotypes accounting for 53% of the isolates tested; 19% of the isolates were untypeable. Only fifteen C. coli serotypes were identified, with three serotypes accounting for 69% of the isolates. This scheme provides a baseline for epidemiological studies of C. jejuni and C. coli.

  14. DNA sequence heterogeneity of Campylobacter jejuni CJIE4 prophages and expression of prophage genes.

    PubMed

    Clark, Clifford G; Chong, Patrick M; McCorrister, Stuart J; Mabon, Philip; Walker, Matthew; Westmacott, Garrett R

    2014-01-01

    Campylobacter jejuni carry temperate bacteriophages that can affect the biology or virulence of the host bacterium. Known effects include genomic rearrangements and resistance to DNA transformation. C. jejuni prophage CJIE1 shows sequence variability and variability in the content of morons. Homologs of the CJIE1 prophage enhance both adherence and invasion to cells in culture and increase the expression of a specific subset of bacterial genes. Other C. jejuni temperate phages have so far not been well characterized. In this study we describe investigations into the DNA sequence variability and protein expression in a second prophage, CJIE4. CJIE4 sequences were obtained de novo from DNA sequencing of five C. jejuni isolates, as well as from whole genome sequences submitted to GenBank by other research groups. These CJIE4 DNA sequences were heterogenous, with several different insertions/deletions (indels) in different parts of the prophage genome. Two variants of a 3-4 kb region inserted within CJIE4 had different gene content that distinguished two major conserved CJIE4 prophage families. Additional indels were detected throughout the prophage. Detection of proteins in the five isolates characterized in our laboratory in isobaric Tags for Relative and Absolute Quantitation (iTRAQ) experiments indicated that prophage proteins within each of the two large indel variants were expressed during growth of the bacteria on Mueller Hinton agar plates. These proteins included the extracellular DNase associated with resistance to DNA transformation and prophage repressor proteins. Other proteins associated with known or suspected roles in prophage biology were also expressed from CJIE4, including capsid protein, the phage integrase, and MazF, a type II toxin-antitoxin system protein. Together with the results previously obtained for the CJIE1 prophage these results demonstrate that sequence variability and expression of moron genes are both general properties of temperate

  15. Evaluation of fecal calprotectin in Campylobacter concisus and Campylobacter jejuni/coli gastroenteritis.

    PubMed

    Nielsen, Hans Linde; Engberg, Jørgen; Ejlertsen, Tove; Nielsen, Henrik

    2013-05-01

    Calprotectin (CP) is a calcium-binding cytosolic neutrophil protein and the concentration in feces reflects the migration of neutrophils into the gut lumen. Testing for fecal CP (f-CP) in patients with negative cultures for enteric pathogens is widely accepted as a useful screening tool for identifying patients who are most likely to benefit from endoscopy for suspected inflammatory bowel disease (IBD) with the assumption that a negative f-CP is compatible with a functional disorder. Campylobacter concisus has recently been reported to have a high incidence in the Danish population almost equal to Campylobacter jejuni and Campylobacter coli and has been reported to cause prolonged watery diarrhea. However, isolation of C. concisus from feces requires the filter method in a hydrogen-enriched microaerobic atmosphere, which is not commonly used in the laboratory, and the diagnosis may consequently be missed. The aim of this study was to evaluate the f-CP levels, as a marker for the intestinal inflammation in C. jejuni/coli- and C. concisus-infected patients. The authors found a high concentration of f-CP (median 631: IQR 221-1274) among 140 patients with C. jejuni/coli infection, whereas the f-CP level among 99 C. concisus-infected patients was significantly lower (median 53: IQR 20-169). The data correlate to the severe inflammatory gastroenteritis seen in patients infected with C. jejuni/coli, whereas C. concisus-infected patients have a much lower intestinal inflammation which could be compared with viral gastroenteritis. Nevertheless, clinicians should be aware of C. concisus infection, especially in patients with prolonged mild diarrhea, in the differential diagnosis to IBD.

  16. Evaluation of fecal calprotectin in Campylobacter concisus and Campylobacter jejuni/coli gastroenteritis.

    PubMed

    Nielsen, Hans Linde; Engberg, Jørgen; Ejlertsen, Tove; Nielsen, Henrik

    2013-05-01

    Calprotectin (CP) is a calcium-binding cytosolic neutrophil protein and the concentration in feces reflects the migration of neutrophils into the gut lumen. Testing for fecal CP (f-CP) in patients with negative cultures for enteric pathogens is widely accepted as a useful screening tool for identifying patients who are most likely to benefit from endoscopy for suspected inflammatory bowel disease (IBD) with the assumption that a negative f-CP is compatible with a functional disorder. Campylobacter concisus has recently been reported to have a high incidence in the Danish population almost equal to Campylobacter jejuni and Campylobacter coli and has been reported to cause prolonged watery diarrhea. However, isolation of C. concisus from feces requires the filter method in a hydrogen-enriched microaerobic atmosphere, which is not commonly used in the laboratory, and the diagnosis may consequently be missed. The aim of this study was to evaluate the f-CP levels, as a marker for the intestinal inflammation in C. jejuni/coli- and C. concisus-infected patients. The authors found a high concentration of f-CP (median 631: IQR 221-1274) among 140 patients with C. jejuni/coli infection, whereas the f-CP level among 99 C. concisus-infected patients was significantly lower (median 53: IQR 20-169). The data correlate to the severe inflammatory gastroenteritis seen in patients infected with C. jejuni/coli, whereas C. concisus-infected patients have a much lower intestinal inflammation which could be compared with viral gastroenteritis. Nevertheless, clinicians should be aware of C. concisus infection, especially in patients with prolonged mild diarrhea, in the differential diagnosis to IBD. PMID:23448294

  17. High frequency genetic variation of purine biosynthesis genes is a mechanism of success in Campylobacter jejuni

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Phenotypic variation is prevalent among progeny of the zoonotic pathogen Campylobacter jejuni, the leading agent of enterocolitis in the developed world. Heterogeneity bestows increased survival to bacterial populations because variable phenotypes ensure some cells will be protected against future s...

  18. Alternative bacteriophage life cycles: the carrier state of Campylobacter jejuni.

    PubMed

    Siringan, Patcharin; Connerton, Phillippa L; Cummings, Nicola J; Connerton, Ian F

    2014-01-01

    Members of the genus Campylobacter are frequently responsible for human enteric disease, often through consumption of contaminated poultry products. Bacteriophages are viruses that have the potential to control pathogenic bacteria, but understanding their complex life cycles is key to their successful exploitation. Treatment of Campylobacter jejuni biofilms with bacteriophages led to the discovery that phages had established a relationship with their hosts typical of the carrier state life cycle (CSLC), where bacteria and bacteriophages remain associated in equilibrium. Significant phenotypic changes include improved aerotolerance under nutrient-limited conditions that would confer an advantage to survive in extra-intestinal environments, but a lack in motility eliminated their ability to colonize chickens. Under these circumstances, phages can remain associated with a compatible host and continue to produce free virions to prospect for new hosts. Moreover, we demonstrate that CSLC host bacteria can act as expendable vehicles for the delivery of bacteriophages to new host bacteria within pre-colonized chickens. The CSLC represents an important phase in the ecology of Campylobacter bacteriophage. PMID:24671947

  19. Alternative bacteriophage life cycles: the carrier state of Campylobacter jejuni.

    PubMed

    Siringan, Patcharin; Connerton, Phillippa L; Cummings, Nicola J; Connerton, Ian F

    2014-01-01

    Members of the genus Campylobacter are frequently responsible for human enteric disease, often through consumption of contaminated poultry products. Bacteriophages are viruses that have the potential to control pathogenic bacteria, but understanding their complex life cycles is key to their successful exploitation. Treatment of Campylobacter jejuni biofilms with bacteriophages led to the discovery that phages had established a relationship with their hosts typical of the carrier state life cycle (CSLC), where bacteria and bacteriophages remain associated in equilibrium. Significant phenotypic changes include improved aerotolerance under nutrient-limited conditions that would confer an advantage to survive in extra-intestinal environments, but a lack in motility eliminated their ability to colonize chickens. Under these circumstances, phages can remain associated with a compatible host and continue to produce free virions to prospect for new hosts. Moreover, we demonstrate that CSLC host bacteria can act as expendable vehicles for the delivery of bacteriophages to new host bacteria within pre-colonized chickens. The CSLC represents an important phase in the ecology of Campylobacter bacteriophage.

  20. [Enzymatic activities of Campylobacter jejuni, C. coli, and C. lari].

    PubMed

    Colomina, J; Villar, J; Buesa, J; Borras, R

    1997-01-01

    Susceptibility to nalidixic acid has been considered for a long time as an important test in the identification of the different species of the genus Campylobacter. However, due to the increasing development of resistance new laboratory tests are needed to carry out an accurate identification to the species level in combination with other typing schemes. We have evaluated the enzymatic activity of 180 Campylobacter spp. strains isolated from clinical specimens performing the Api ZYM system (bioMérieux) in order to assay whether different enzymatic patterns could help to characterize these microorganisms. Thirteen of the 19 enzymatic activities detected by the system (lipase-C14, valine arylamidase, cystine arylamidase, trypsin, chymiotrypsin, alpha-galactosidase, beta-galactosidase, beta-glucuronidase, alpha-glucosidase, beta-glucosidase, N-acetyl-beta-glucosaminidase, alpha-manosidase and alpha-fucosidase) were negative for all the strains tested, whereas 3 enzymes (alkaline phosphatase, acid phosphatase and naphtol-A-S-BI-phosphohydrolase) were detected in 96.6% of the strains. Esterase-C1, esterase lipase-C8 and leucin arylamidase showed a variable reactivity depending on isolates. Enzymatic activity patterns clearly differentiate all the C. lari strains from other Campylobacter species. No significant differences were detected among the enzymatic activities of C. jejuni and C. coli strains. Our study suggests that the Api ZYM system is easy to perform and a valuable method to be applied in the characterization of the campylobacteria as a complement to other biotyping and serotyping schemes.

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The antibacterial effect of ZnO nanoparticles on Campylobacter jejuni was investigated for cell growth inhibition and inactivation. The results showed that C. jejuni was extremely sensitive to the treatment of ZnO nanoparticles. The minimal inhibitory concentration (MIC) of ZnO nanoparticles to C. j...

  2. Bacteriocins reduce Campylobacter jejuni colonization while bacteria producing bacteriocins are ineffective

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Broiler chickens are widely considered an important source for human exposure to Campylobacter jejuni because of the high numbers found colonizing the chicken gut and the consequent contamination of processed carcasses. We hoped to intervene in gut colonization by C. jejuni using a defined probioti...

  3. Caught in the Act: In Vivo Development of Macrolide Resistance to Campylobacter jejuni Infection ▿

    PubMed Central

    Lindow, J. C.; Poly, F.; Tribble, D. R.; Guerry, P.; Carmolli, M. P.; Baqar, S.; Porter, C. K.; Pierce, K. K.; Darsley, M. J.; Sadigh, K. S.; Dill, E. A.; Kirkpatrick, B. D.

    2010-01-01

    We report the first instance of macrolide resistance developing in vivo following appropriate antibiotic use in a healthy volunteer as a part of a controlled human infection with Campylobacter jejuni. In vivo development of macrolide resistance may be an important contributor to antibiotic resistance in C. jejuni. PMID:20554818

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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...

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The species Campylobacter jejuni (C. jejuni) is considered naturally competent for DNA uptake and displays strong genetic diversity. Yet, non-transformable strains and several relatively stable clonal lineages exist. In the present study, the molecular mechanism responsible for the non-transformabil...

  6. Development of antimicrobial resistance in Campylobacter jejuni and Campylobacter coli adapted to biocides.

    PubMed

    Mavri, Ana; Smole Možina, Sonja

    2013-01-01

    The potential for adaptive resistance of Campylobacter jejuni and Campylobacter coli after step-wise exposure to increasing sub-inhibitory concentrations of five biocides as triclosan, benzalkonium chloride, cetylpyridinium chloride, chlorhexidine diacetate and trisodium phosphate, was investigated, to identify the mechanisms underlying resistance. The biocide resistance and cross-resistance to the antimicrobials erythromycin and ciprofloxacin, and to sodium dodecyl sulphate, were examined according to the broth microdilution method. The presence of active efflux was studied on the basis of restored sensitivity in the presence of the efflux pump inhibitors phenylalanine-arginine beta-naphthylamide, 1-(1-naphthylmethyl)-piperazine, cyanide 3-chlorophenylhydrazone, verapamil and reserpine. Changes in the outer membrane protein profiles and morphological changes in adapted strains were studied, as compared with the parent strains. Repeated exposure of C. jejuni and C. coli to biocides resulted in partial increases in tolerance to biocides itself, to other biocides and antimicrobial compounds. The developed resistance was stable for up to 10 passages in biocide-free medium. More than one type of active efflux was identified in adapted strains. These adapted strains showed different alterations to their outer membrane protein profiles, along with morphological changes. The data presented here suggest that different mechanisms are involved in adaptation to biocides and that this adaptation is unique to each strain of Campylobacter and does not result from a single species-specific mechanism.

  7. Evaluation of an enzyme immunoassay-based stool antigen test to detect Campylobacter jejuni and Campylobacter coli.

    PubMed

    Tissari, Päivi; Rautelin, Hilpi

    2007-06-01

    An enzyme immunoassay-based antigen test (Ridascreen Campylobacter; R-Biopharm, Darmstadt, Germany) was evaluated for the detection of Campylobacter jejuni and Campylobacter coli in 1050 clinical stool samples as compared with culture on selective medium. After routine inoculation for Salmonella, Shigella, Yersinia, Aeromonas, Plesiomonas, and Campylobacter, the same swab specimens were used for the antigen test. The positivity rate for Campylobacter was 9.3% in culture, and the antigen test gave a sensitivity of 69%. Forty-six stool samples culture-negative for Campylobacter grew other enteropathogens; one (positive for Salmonella sp.) was positive in the antigen test. Of all the 952 Campylobacter culture-negative samples, 830 were negative in the antigen test, giving a specificity of 87%. Almost 5% of the samples showed equivocal antigen test results. If the moderate sensitivity of the antigen test was due to a low sensitivity of culture or receiving the stool samples in transportation tubes remains to be studied.

  8. Rapid identification of thermotolerant Campylobacter jejuni, Campylobacter coli, Campylobacter lari, and Campylobacter upsaliensis from various geographic locations by a GTPase-based PCR-reverse hybridization assay.

    PubMed

    van Doorn, L J; Verschuuren-van Haperen, A; Burnens, A; Huysmans, M; Vandamme, P; Giesendorf, B A; Blaser, M J; Quint, W G

    1999-06-01

    Recently, a gene from Campylobacter jejuni encoding a putative GTPase was identified. Based on two semiconserved GTP-binding sites encoded within this gene, PCR primers were selected that allow amplification of a 153-bp fragment from C. jejuni, C. coli, C. lari, and C. upsaliensis. Sequence analysis of these PCR products revealed consistent interspecies variation, which allowed the definition of species-specific probes for each of the four thermotolerant Campylobacter species. Multiple probes were used to develop a line probe assay (LiPA) that permits analysis of PCR products by a single reverse hybridization step. A total of 320 reference strains and clinical isolates from various geographic origins were tested by the GTP-based PCR-LiPA. The PCR-LiPA is highly specific in comparison with conventional identification methods, including biochemical and whole-cell protein analyses. In conclusion, a simple method has been developed for rapid and highly specific identification of thermotolerant Campylobacter species.

  9. Novel plasmid conferring kanamycin and tetracycline resistance in turkey-derived Campylobacter jejuni strain 11601MD

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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...

  10. New World Monkey Aotus nancymae as a Model for Campylobacter jejuni Infection and Immunity

    PubMed Central

    Jones, Franca R.; Baqar, Shahida; Gozalo, Alfonso; Nunez, Gladys; Espinoza, Nereyda; Reyes, Sharina M.; Salazar, Milagros; Meza, Rina; Porter, Chad K.; Walz, Stephen E.

    2006-01-01

    Three groups of six monkeys (Aotus nancymae) each were inoculated intragastrically with increasing doses of Campylobacter jejuni. Infection resulted in fecal colonization (100% of monkeys), dose-related diarrhea, and robust immune responses. Colonization duration and diarrhea rate were reduced upon secondary challenge. A. nancymae may be useful for studying anti-Campylobacter vaccine efficacy. PMID:16369042

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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 ...

  12. Chemically defined media for auxotyping of Campylobacter jejuni.

    PubMed

    Dickgiesser, N; Czylwik, D

    1985-08-01

    A set of chemically defined media has been developed for the cultivation of Campylobacter jejuni strains of human origin. A minimal medium, a complete medium and 5 different nutrient-deficient media (NDM1-NDM5) are described. Some of the strains investigated required L-methionine(lacking in NDM1), L-cystine and L-cysteine (NDM2), K2HPO4 (NDM 3), KH2PO4 (NDM4) and NAD, thiamine and calcium pantothenate (NDM5). 57.7% of the strains investigated required L-methionine. The strains grew at pH 6.6-7.7. The media described are not suitable for C. intestinalis. PMID:4060921

  13. Enterotoxin production and serogroups of Campylobacter jejuni and Campylobacter coli from patients with diarrhea and from healthy laying hens.

    PubMed

    Lindblom, G B; Kaijser, B; Sjögren, E

    1989-06-01

    Enterotoxin production, a possible virulence factor, was determined in Campylobacter jejuni and Campylobacter coli by two different techniques, the CHO cell test and the GM1 enzyme-linked immunosorbent assay. The frequency of enterotoxigenic Campylobacter strains was 32% in strains from both humans with acute enteritis and healthy laying hens, as measured by the CHO cell test. The CHO cell test was significantly more sensitive than the GM1 enzyme-linked immunosorbent assay in the detection of enterotoxigenic strains. Enterotoxin production was compared with the presence of heat-stable and heat-labile antigens. There was no significant correlation between enterotoxin production and serogroups for C. jejuni or C. coli. The difference in enterotoxigenicity between C. jejuni (34.1%) and C. coli (21.9%) was not significant.

  14. An Improved Culture Method for Selective Isolation of Campylobacter jejuni from Wastewater.

    PubMed

    Kim, Jinyong; Oh, Euna; Banting, Graham S; Braithwaite, Shannon; Chui, Linda; Ashbolt, Nicholas J; Neumann, Norman F; Jeon, Byeonghwa

    2016-01-01

    Campylobacter jejuni is one of the leading foodborne pathogens worldwide. C. jejuni is isolated from a wide range of foods, domestic animals, wildlife, and environmental sources. The currently available culture-based isolation methods are not highly effective for wastewater samples due to the low number of C. jejuni in the midst of competing bacteria. To detect and isolate C. jejuni from wastewater samples, in this study, we evaluated a few different enrichment conditions using five different antibiotics (i.e., cefoperazone, vancomycin, trimethoprim, polymyxin B, and rifampicin), to which C. jejuni is intrinsically resistant. The selectivity of each enrichment condition was measured with C t value using quantitative real-time PCR, and multiplex PCR to determine Campylobacter species. In addition, the efficacy of Campylobacter isolation on different culture media after selective enrichment was examined by growing on Bolton and Preston agar plates. The addition of polymyxin B, rifampicin, or both to the Bolton selective supplements enhanced the selective isolation of C. jejuni. The results of 16S rDNA sequencing also revealed that Enterococcus spp. and Pseudomonas aeruginosa are major competing bacteria in the enrichment conditions. Although it is known to be difficult to isolate Campylobacter from samples with heavy contamination, this study well exhibited that the manipulation of antibiotic selective pressure improves the isolation efficiency of fastidious Campylobacter from wastewater. PMID:27617011

  15. An Improved Culture Method for Selective Isolation of Campylobacter jejuni from Wastewater

    PubMed Central

    Kim, Jinyong; Oh, Euna; Banting, Graham S.; Braithwaite, Shannon; Chui, Linda; Ashbolt, Nicholas J.; Neumann, Norman F.; Jeon, Byeonghwa

    2016-01-01

    Campylobacter jejuni is one of the leading foodborne pathogens worldwide. C. jejuni is isolated from a wide range of foods, domestic animals, wildlife, and environmental sources. The currently available culture-based isolation methods are not highly effective for wastewater samples due to the low number of C. jejuni in the midst of competing bacteria. To detect and isolate C. jejuni from wastewater samples, in this study, we evaluated a few different enrichment conditions using five different antibiotics (i.e., cefoperazone, vancomycin, trimethoprim, polymyxin B, and rifampicin), to which C. jejuni is intrinsically resistant. The selectivity of each enrichment condition was measured with Ct value using quantitative real-time PCR, and multiplex PCR to determine Campylobacter species. In addition, the efficacy of Campylobacter isolation on different culture media after selective enrichment was examined by growing on Bolton and Preston agar plates. The addition of polymyxin B, rifampicin, or both to the Bolton selective supplements enhanced the selective isolation of C. jejuni. The results of 16S rDNA sequencing also revealed that Enterococcus spp. and Pseudomonas aeruginosa are major competing bacteria in the enrichment conditions. Although it is known to be difficult to isolate Campylobacter from samples with heavy contamination, this study well exhibited that the manipulation of antibiotic selective pressure improves the isolation efficiency of fastidious Campylobacter from wastewater.

  16. An Improved Culture Method for Selective Isolation of Campylobacter jejuni from Wastewater

    PubMed Central

    Kim, Jinyong; Oh, Euna; Banting, Graham S.; Braithwaite, Shannon; Chui, Linda; Ashbolt, Nicholas J.; Neumann, Norman F.; Jeon, Byeonghwa

    2016-01-01

    Campylobacter jejuni is one of the leading foodborne pathogens worldwide. C. jejuni is isolated from a wide range of foods, domestic animals, wildlife, and environmental sources. The currently available culture-based isolation methods are not highly effective for wastewater samples due to the low number of C. jejuni in the midst of competing bacteria. To detect and isolate C. jejuni from wastewater samples, in this study, we evaluated a few different enrichment conditions using five different antibiotics (i.e., cefoperazone, vancomycin, trimethoprim, polymyxin B, and rifampicin), to which C. jejuni is intrinsically resistant. The selectivity of each enrichment condition was measured with Ct value using quantitative real-time PCR, and multiplex PCR to determine Campylobacter species. In addition, the efficacy of Campylobacter isolation on different culture media after selective enrichment was examined by growing on Bolton and Preston agar plates. The addition of polymyxin B, rifampicin, or both to the Bolton selective supplements enhanced the selective isolation of C. jejuni. The results of 16S rDNA sequencing also revealed that Enterococcus spp. and Pseudomonas aeruginosa are major competing bacteria in the enrichment conditions. Although it is known to be difficult to isolate Campylobacter from samples with heavy contamination, this study well exhibited that the manipulation of antibiotic selective pressure improves the isolation efficiency of fastidious Campylobacter from wastewater. PMID:27617011

  17. Bidirectional transcription of lipooligosaccharide synthesis genes from Campylobacter jejuni.

    PubMed

    Phongsisay, Vongsavanh; Fry, Benjamin N

    2007-10-01

    The lipooligosaccharide (LOS) molecules of Campylobacter jejuni are involved in virulence and induction of the Guillain-Barré syndrome (GBS). This study analysed the transcription of the LOS synthesis genes from the GBS-inducing C. jejuni strain HB 93-13 under microaerobic conditions. Fourteen consecutive genes Cj1132c, waaC, htrB, wlaNC, wlaND, cgtA, cgtB, cstII, neuB, neuC, neuA, wlaVA, wlaQA, and waaF were included. The results of rapid amplification of cDNA ends and single-stranded ligation of complementary ends showed initiation sites with potential promoter regions on both DNA strands in the Cj1132c/waaC, cgtB/cstII, and wlaQA/waaF strand-switch regions. Other termini without recognisable promoter region were also found throughout the LOS gene cluster, suggesting a low specificity of the polymerase during transcription. In addition, all gene junction regions were cloned into the shuttle vector pMW10 carrying the promoterless lacZ gene to identify functional promoter sites. Bidirectional active promoters were found in the strand-switch regions. The results of RT-PCR and cDNA blotting indicated that transcriptional linkage occurred between different operons, indicating a lack of transcription termination within the LOS gene cluster. Moreover, the results of semi-quantitative RT-PCR and real-time RT-PCR showed that both DNA strands were transcribed but transcription of the coding strand was at a higher rate. The results presented here provide an insight into transcription of the LOS synthesis gene cluster of C. jejuni.

  18. Campylobacter jejuni PflB is required for motility and colonisation of the chicken gastrointestinal tract.

    PubMed

    Kanji, Alpa; Jones, Michael A; Maskell, Duncan J; Grant, Andrew J

    2015-12-01

    Campylobacter jejuni is the leading cause of foodborne bacterial gastroenteritis worldwide. Although the mechanisms by which C. jejuni causes disease are not completely understood, the presence of functional flagella appears to be required for colonisation of the gastrointestinal tract of humans and animals. Therefore much attention has been given to understanding the synthesis and role of flagella in C. jejuni. In this study we report insights into the function of PflB that is essential for Campylobacter motility. We have explored the function of this gene by constructing deletion mutants in C. jejuni strains NCTC11168 and M1, in the genes cj0390 and CJM1_0368, respectively. The mutants were non-motile yet assembled flagella that appeared structurally identical to the wild type. Furthermore the protein is required for C. jejuni colonisation of caeca in a two-week old chicken colonisation model.

  19. Insights into Campylobacter jejuni colonization and enteritis using a novel infant rabbit model

    PubMed Central

    Shang, Yuwei; Ren, Fangzhe; Song, Zhaojun; Li, Qiuchun; Zhou, Xiaohui; Wang, Xiaobo; Xu, Zhonglan; Bao, Guangyu; Wan, Ting; Lei, Tianyao; Wang, Nan; Jiao, Xin-an; Huang, Jinlin

    2016-01-01

    A lack of relevant disease models for Campylobacter jejuni has long been an obstacle to research into this common enteric pathogen. Here we used an infant rabbit to study C. jejuni infection, which enables us to define several previously unknown but key features of the organism. C. jejuni is capable of systemic invasion in the rabbit, and developed a diarrhea symptom that mimicked that observed in many human campylobacteriosis. The large intestine was the most consistently colonized site and produced intestinal inflammation, where specific cytokines were induced. Genes preferentially expressed during C. jejuni infection were screened, and acs, cj1385, cj0259 seem to be responsible for C. jejuni invasion. Our results demonstrates that the infant rabbit can be used as an alternative experimental model for the study of diarrheagenic Campylobacter species and will be useful in exploring the pathogenesis of other related pathogens. PMID:27357336

  20. Diversity of pulsed-field gel electrophoresis profiles of Campylobacter jejuni and Campylobacter coli from broiler chickens in France.

    PubMed

    Denis, M; Rose, V; Huneau-Salaün, A; Balaine, L; Salvat, G

    2008-08-01

    In 2003 to 2004, 26 free-range broilers flocks excreting Campylobacter were studied for identification of Campylobacter species and genotype diversity. Seventeen flocks were sampled at the end of the indoor rearing period and 9 before departure to the slaughterhouse after access to an open area. Out of 513 isolates, 315 were identified as Campylobacter jejuni and 198 as Campylobacter coli. Pulsed-field gel electrophoresis analysis revealed 35 genotypes for C. jejuni and 43 genotypes for C. coli; 38.4% of the isolates were clustered into 16 genetic groups. This kind of poultry production system is characterized by a large number of Campylobacter coli isolates. Flocks sampled during the indoor phase were predominantly contaminated by C. jejuni, whereas those sampled during warm months were predominantly contaminated by C. coli. The Campylobacter population was genetically highly diverse: multiple genotypes were detected in a single flock. Six flocks were each found to harbor a mixture of genotypes; these isolates were genetically closely related and were grouped into clusters of at least 80% genetic similarity. Isolates with genotypes found in different flocks and strains, but sharing the same genetic clusters, were detected in different farms and at different times in the year. Flocks sampled during the indoor rearing period and when farmers use fresh farm-made litter were associated with a small number of clusters. In conclusion, Campylobacter species were genetically highly diverse. Our findings suggest the presence of genomic rearrangements in Campylobacter colonizing the chick intestine and that some Campylobacter strains are adapted to poultry. In addition, the species diversity in the flocks was affected by season and cloistering measures. Litter and exposure to an open area were likely sources of flock Campylobacter contamination.

  1. Antimicrobial susceptibility profiles and molecular typing of Campylobacter jejuni and Campylobacter coli isolates from ducks in South Korea.

    PubMed

    Wei, Bai; Cha, Se-Yeoun; Kang, Min; Roh, Jae-Hee; Seo, Hye-Suk; Yoon, Ran-Hee; Jang, Hyung-Kwan

    2014-12-01

    Campylobacter is a food-borne zoonotic pathogen that causes human gastroenteritis worldwide. Campylobacter bacteria are commensal in the intestines of many food production animals, including ducks and chickens. The objective of the study was to determine the prevalence of Campylobacter species in domestic ducks, and the agar dilution method was used to determine resistance of the isolates to eight antibiotics. In addition, multilocus sequence typing (MLST) was performed to determine the sequence types (STs) of selected Campylobacter isolates. Between May and September 2012, 58 duck farms were analyzed, and 56 (96.6%) were positive for Campylobacter. Among the isolates, 82.1% were Campylobacter jejuni, 16.1% were C. coli, and one was unidentified by PCR. Of the 46 C. jejuni isolates, 87.0%, 10.9%, and 21.7% were resistant to ciprofloxacin, erythromycin, and azithromycin, respectively. Among the C. coli isolates, all 9 strains were resistant to ampicillin, and 77.8% and 33.3% were resistant to ciprofloxacin and azithromycin, respectively. The majority of the Campylobacter isolates were classified as multidrug resistant. Twenty-eight STs were identified, including 20 STs for C. jejuni and 8 STs for C. coli. The most common clonal complexes in C. jejuni were the ST-21 complex and the ST-45 complex, while the ST-828 complex predominated in C. coli. The majority of isolates were of STs noted in ducks and humans from earlier studies, along with seven STs previously associated only with human disease. These STs overlapped between duck and human isolates, indicating that Campylobacter isolates from ducks should be considered potential sources of human infection.

  2. Effect of clavulanic acid on susceptibility of Campylobacter jejuni and Campylobacter coli to eight beta-lactam antibiotics.

    PubMed Central

    Gaudreau, C L; Lariviere, L A; Lauzer, J C; Turgeon, F F

    1987-01-01

    The effect of clavulanic acid on the susceptibility of 32 strains of Campylobacter jejuni and Campylobacter coli to eight beta-lactam agents was studied. Almost all strains tested became susceptible to amoxicillin and ticarcillin with 1 microgram of clavulanic acid per ml. This compound had little or no effect on susceptibility to penicillin G, cephalothin, cefamandole, and cefoxitin. Clavulanic acid had a marginal effect on cefotaxime and moxalactam susceptibility. PMID:3619428

  3. Complete genome sequence of UV-resistant Campylobacter jejuni RM3194, including an 81.08-kilobase plasmid

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Campylobacter jejuni strain RM3194 was originally isolated from a human with enteritis and contains a novel 81,079-bp plasmid. RM3194 has exhibited superior survival compared to other Campylobacter jejuni strains when challenged with UV light. The chromosome of RM3194 was determined to be 1,651,18...

  4. COMPARATIVE ASSESSMENT OF STANDARD CULTURE AND REAL-TIME PCR TO DETECT CAMPYLOBACTER JEJUNI IN RETAIL CHICKEN SAMPLES

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Contamination of poultry by Campylobacter is a significant source of human diarrheal illness. Conventional bacteriological methods to detect and speciate Campylobacter jejuni (C. jejuni) from chicken samples are labor-intensive and time-consuming. The purpose of this study was to compare standard c...

  5. A novel mouse model of Campylobacter jejuni gastroenteritis reveals key pro-inflammatory and tissue protective roles for Toll-like receptor signaling during infection.

    PubMed

    Stahl, Martin; Ries, Jenna; Vermeulen, Jenny; Yang, Hong; Sham, Ho Pan; Crowley, Shauna M; Badayeva, Yuliya; Turvey, Stuart E; Gaynor, Erin C; Li, Xiaoxia; Vallance, Bruce A

    2014-07-01

    Campylobacter jejuni is a major source of foodborne illness in the developed world, and a common cause of clinical gastroenteritis. Exactly how C. jejuni colonizes its host's intestines and causes disease is poorly understood. Although it causes severe diarrhea and gastroenteritis in humans, C. jejuni typically dwells as a commensal microbe within the intestines of most animals, including birds, where its colonization is asymptomatic. Pretreatment of C57BL/6 mice with the antibiotic vancomycin facilitated intestinal C. jejuni colonization, albeit with minimal pathology. In contrast, vancomycin pretreatment of mice deficient in SIGIRR (Sigirr(-/-)), a negative regulator of MyD88-dependent signaling led to heavy and widespread C. jejuni colonization, accompanied by severe gastroenteritis involving strongly elevated transcription of Th1/Th17 cytokines. C. jejuni heavily colonized the cecal and colonic crypts of Sigirr(-/-) mice, adhering to, as well as invading intestinal epithelial cells. This infectivity was dependent on established C. jejuni pathogenicity factors, capsular polysaccharides (kpsM) and motility/flagella (flaA). We also explored the basis for the inflammatory response elicited by C. jejuni in Sigirr(-/-) mice, focusing on the roles played by Toll-like receptors (TLR) 2 and 4, as these innate receptors were strongly stimulated by C. jejuni. Despite heavy colonization, Tlr4(-/-)/Sigirr(-/-) mice were largely unresponsive to infection by C. jejuni, whereas Tlr2(-/-)/Sigirr(-/-) mice developed exaggerated inflammation and pathology. This indicates that TLR4 signaling underlies the majority of the enteritis seen in this model, whereas TLR2 signaling had a protective role, acting to promote mucosal integrity. Furthermore, we found that loss of the C. jejuni capsule led to increased TLR4 activation and exaggerated inflammation and gastroenteritis. Together, these results validate the use of Sigirr(-/-) mice as an exciting and relevant animal model for

  6. Loop-Mediated Isothermal Amplification (LAMP) for Detection of Campylobacter jejuni and C. coli in Thai Children with Diarrhea.

    PubMed

    Pham, Ngan Thi Kim; Trinh, Quang Duy; Khamrin, Pattara; Ukarapol, Nuthapong; Kongsricharoern, Tipachan; Yamazaki, Wataru; Komine-Aizawa, Shihoko; Okitsu, Shoko; Maneekarn, Niwat; Hayakawa, Satoshi; Ushijima, Hiroshi

    2015-01-01

    Campylobacter species are common causes of bacterial diarrhea, and Campylobacter jejuni and C. coli are known as the predominant causative agents in humans. Recent studies suggested that loop-mediated isothermal amplification (LAMP) is an efficient and practical tool for rapid detection of C. jejuni and C. coli in clinical samples. We used LAMP to screen 151 stool samples for Campylobacter; these samples were collected in 2012 from Thai children with diarrhea. The PCR method discriminated C. jejuni and C. coli among the detected Campylobacter strains; these species were subjected to sequencing of the hipO gene (in C. jejuni) or the ask gene (in C. coli). The results suggest that the prevalence of Campylobacter infection among Thai children with diarrhea is 8.6%, and C. jejuni is the most prevalent species.

  7. Narrow-spectrum inhibitors of Campylobacter jejuni flagellar expression and growth.

    PubMed

    Johnson, Jeremiah G; Yuhas, Caroline; McQuade, Thomas J; Larsen, Martha J; DiRita, Victor J

    2015-07-01

    Campylobacter jejuni is a major cause of food-borne illness due to its ability to reside within the gastrointestinal tracts of chickens. Multiple studies have identified the flagella of C. jejuni as a major determinant of chicken colonization. An inhibitor screen of approximately 147,000 small molecules was performed to identify compounds that are able to inhibit flagellar expression in a reporter strain of C. jejuni. Several compounds that modestly inhibited motility of wild-type C. jejuni in standard assays were identified, as were a number of small molecules that robustly inhibited C. jejuni growth, in vitro. Examination of similar bacterial screens found that many of these small molecules inhibited only the growth of C. jejuni. Follow-up assays demonstrated inhibition of other strains of C. jejuni and Campylobacter coli but no inhibition of the closely related Helicobacter pylori. The compounds were determined to be bacteriostatic and nontoxic to eukaryotic cells. Preliminary results from a day-of-hatch chick model of colonization suggest that at least one of the compounds demonstrates promise for reducing Campylobacter colonization loads in vivo, although further medicinal chemistry may be required to enhance bioavailability.

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

    PubMed Central

    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

    2016-01-01

    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. PMID:27109214

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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...

  10. Systemic response to Campylobacter jejuni infection by profiling gene transcription in the spleens of two genetic lines of chickens

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Campylobacter jejuni (C. jejuni) is a leading cause of human bacterial enteritis worldwide. To understand the systemic molecular response mechanisms to C. jejuni infection in chickens, total splenic RNA was isolated and applied to a whole genome chicken microarray for comparison between infected (I...

  11. Prevalence of Type VI Secretion System in Spanish Campylobacter jejuni Isolates.

    PubMed

    Ugarte-Ruiz, M; Stabler, R A; Domínguez, L; Porrero, M C; Wren, B W; Dorrell, N; Gundogdu, O

    2015-11-01

    Infections from Campylobacter jejuni pose a serious public health problem and are now considered the leading cause of foodborne bacterial gastroenteritis throughout the world. Sequencing of C. jejuni genomes has previously allowed a number of loci to be identified, which encode virulence factors that aid survival and pathogenicity. Recently, a Type VI secretion system (T6SS) consisting of 13 conserved genes was described in C. jejuni strains and recognised to promote pathogenicity and adaptation to the environment. In this study, we determined the presence of this T6SS in 63 Spanish C. jejuni isolates from the food chain and urban effluents using whole-genome sequencing. Our findings demonstrated that nine (14%) strains harboured the 13 ORFs found in prototype strain C. jejuni 108. Further studies will be necessary to determine the prevalence and importance of T6SS-positive C. jejuni strains.

  12. 23S rRNA gene mutations contributing to macrolide resistance in Campylobacter jejuni and Campylobacter coli

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Operon specific 23S rRNA mutations affecting minimum inhibitory concentrations (MICs) of macrolides (erythromycin [ERY], azithromycin [AZM], tylosin [TYL]) and a lincosamide (clindamycin [CLI]) were examined in a collection of Campylobacter jejuni and C. coli isolates. The three copies of the Campy...

  13. Risk factors for indigenous Campylobacter jejuni and Campylobacter coli infections in The Netherlands: a case-control study.

    PubMed

    Doorduyn, Y; Van Den Brandhof, W E; Van Duynhoven, Y T H P; Breukink, B J; Wagenaar, J A; Van Pelt, W

    2010-10-01

    A case-control study comprising 1315 Campylobacter jejuni cases, 121 Campylobacter coli cases and 3409 frequency-matched controls was conducted in The Netherlands in 2002-2003. Risk factors for both C. jejuni and C. coli enteritis were consumption of undercooked meat and barbecued meat, ownership of cats and use of proton pump inhibitors. Consumption of chicken was a predominant risk factor for C. jejuni enteritis, but many additional risk factors were identified. Unique risk factors for C. coli infections were consumption of game and tripe, and swimming. Contact with farm animals and persons with gastroenteritis were predominant risk factors for C. jejuni enteritis in young children (0-4 years). Important risk factors for the elderly (>or=60 years) were eating in a restaurant, use of proton pump inhibitors and having a chronic intestinal illness. Consumption of chicken in spring, steak tartare in autumn and winter and barbecued meat in rural areas showed strong associations with C. jejuni infections. This study illustrates that important differences in risk factors exist for different Campylobacter spp. and these may differ dependent on age, season or degree of urbanization. PMID:20223048

  14. 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.

  15. 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

  16. Chicken juice enhances surface attachment and biofilm formation of Campylobacter jejuni.

    PubMed

    Brown, Helen L; Reuter, Mark; Salt, Louise J; Cross, Kathryn L; Betts, Roy P; van Vliet, Arnoud H M

    2014-11-01

    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. 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.

  18. Characterization of Campylobacter jejuni and Campylobacter coli genotypes in poultry flocks by restriction fragment length polymorphism (RFLP) analysis.

    PubMed

    Carreira, Ana Cláudia; Cunha, Mónica V

    2015-01-01

    We describe a simple, rapid, and discriminatory methodology that allows the routine molecular characterization of Campylobacter jejuni and Campylobacter coli isolates. The proposed approach is built on one of the earliest and simplest molecular typing methods ever, consisting on the analysis of the fragments of different lengths generated by digestion of homologous DNA sequences with specific restriction endonucleases, a process known as restriction fragment length polymorphism (RFLP) analysis. The strategy underneath the workflow reported here is meant to explore the polymorphisms of Campylobacter spp. flaA gene (flaA-RFLP) that allows the local investigation of the genetic diversity and distribution of C. coli and C. jejuni isolates from different sources, namely, chickens' caeca. Although not appropriate for global and long-term epidemiological studies as a single approach, flaA-RFLP analysis can be very useful in surveys limited in space and time and, for specific epidemiological settings, an alternative to more modern and resource-demanding techniques.

  19. Use of Caprylic Acid in Broiler Chickens: Effect on Campylobacter jejuni.

    PubMed

    Hovorková, Petra; Skřivanová, Eva

    2015-08-01

    The effect of caprylic acid (CA) on Campylobacter jejuni in chickens was evaluated using two approaches: dietary supplementation or surface treatment of chilled chicken carcasses. To analyze the dietary effect of CA, individually housed broiler chickens (n = 48) were artificially infected with C. jejuni VFU612 (10(6) colony-forming units [CFU]/bird) on the 21st and 35th days of life. Dietary CA (2.5 and 5 g/kg of feed, fed throughout the entire experiment) significantly decreased C. jejuni shedding (p<0.05). However, the effect only lasted for 3-7 days after infection. The numbers of Campylobacter shed by the positive control birds reached its maximum on the 37th day of life, while on that same day, both Treatment I and Treatment II groups shed significantly lower (p<0.05) numbers of Campylobacter (by 0.8 and 1.8 log10 CFU/g, respectively). Also, peak shedding was delayed by 1 day in both treated groups. After euthanasia of each chicken on the 42nd day of life, no differences in Campylobacter counts in the crop, gizzard, ileum, and cecum were found between the positive control and the treated groups (p>0.05). Surface contamination of the chilled chicken halves was performed with C. jejuni VFU612 (clinical isolate) and CCM6214 (collection strain). Surface treatment with CA at 1.25 and 2.5 mg/mL for 1 min significantly reduced C. jejuni VFU612 contamination of chicken skin (p<0.05) by 0.29-0.53 and 1.14-1.58 log10 CFU/g of skin, respectively. Counts of C. jejuni CCM6214 were reduced by 0.68-1.65 log10 CFU/g of skin). In conclusion, dietary CA affected numbers of C. jejuni in the gastrointestinal contents of chickens, whereas surface treatment reduced C. jejuni contamination in processed chicken carcasses.

  20. Enhanced survival of Campylobacter jejuni in association with wood.

    PubMed

    Boucher, S N; Chamberlain, A H; Adams, M R

    1998-01-01

    Campylobacter jejuni cells, when stressed by aeration of the liquid culture medium, were protected from death when a block of wood was present in the broth, especially at 30 degrees C. Sealing the wood inside a porous membrane showed that access to its physical structure was necessary for the protection of cells and that soluble free-radical scavengers from the wood were not responsible for the protection seen. Insoluble free-radical scavengers associated with the wood were not a significant factor since other porous materials offered protection when their pores were sufficiently small (around 16 micron); and the wood itself needed to be at least 4 mm thick for the effect to be evident. Deeply scored plastic blocks did not enhance the survival of cells in aerated broths. Scanning electron microscopy was used to determine the size of the openings within the wood in relation to the size of the bacterial cells. Thus it was established that the physical structure of the wood, rather than its chemistry, was responsible for its protective effect. The consequences of this findings for the use of wooden cutting boards in food handling are discussed. PMID:9708248

  1. The Intestinal Microbiota Influences Campylobacter jejuni Colonization and Extraintestinal Dissemination in Mice.

    PubMed

    O'Loughlin, Jason L; Samuelson, Derrick R; Braundmeier-Fleming, Andrea G; White, Bryan A; Haldorson, Gary J; Stone, Jennifer B; Lessmann, Jeremy J; Eucker, Tyson P; Konkel, Michael E

    2015-07-01

    Campylobacter jejuni is a leading cause of human foodborne gastroenteritis worldwide. The interactions between this pathogen and the intestinal microbiome within a host are of interest as endogenous intestinal microbiota mediates a form of resistance to the pathogen. This resistance, termed colonization resistance, is the ability of commensal microbiota to prevent colonization by exogenous pathogens or opportunistic commensals. Although mice normally demonstrate colonization resistance to C. jejuni, we found that mice treated with ampicillin are colonized by C. jejuni, with recovery of Campylobacter from the colon, mesenteric lymph nodes, and spleen. Furthermore, there was a significant reduction in recovery of C. jejuni from ampicillin-treated mice inoculated with a C. jejuni virulence mutant (ΔflgL strain) compared to recovery of mice inoculated with the C. jejuni wild-type strain or the C. jejuni complemented isolate (ΔflgL/flgL). Comparative analysis of the microbiota from nontreated and ampicillin-treated CBA/J mice led to the identification of a lactic acid-fermenting isolate of Enterococcus faecalis that prevented C. jejuni growth in vitro and limited C. jejuni colonization of mice. Next-generation sequencing of DNA from fecal pellets that were collected from ampicillin-treated CBA/J mice revealed a significant decrease in diversity of operational taxonomic units (OTUs) compared to that in control (nontreated) mice. Taken together, we have demonstrated that treatment of mice with ampicillin alters the intestinal microbiota and permits C. jejuni colonization. These findings provide valuable insights for researchers using mice to investigate C. jejuni colonization factors, virulence determinants, or the mechanistic basis of probiotics. PMID:25934624

  2. Campylobacter jejuni survival within human epithelial cells is enhanced by the secreted protein CiaI

    PubMed Central

    Buelow, Daelynn R.; Christensen, Jeffrey E.; Neal-McKinney, Jason M.; Konkel, Michael E.

    2011-01-01

    Summary Although it is known that Campylobacter jejuni invade the cells that line the human intestinal tract, the bacterial proteins that enable this pathogen to survive within Campylobacter-containing vacuoles (CCV) have not been identified. Here, we describe the identification and characterization of a protein that we termed CiaI for Campylobacter invasion antigen involved in Intracellular survival. We show that CiaI harbors an amino-terminal type III secretion (T3S) sequence and is secreted from C. jejuni through the flagellar T3S system. In addition, the ciaI mutant was impaired in intracellular survival when compared to a wild-type strain, as judged by the gentamicin-protection assay. Fluorescence microscopy examination of epithelial cells infected with the C. jejuni ciaI mutant revealed that the CCV were more frequently co-localized with Cathepsin D (a lysosomal marker) than the CCV in cells infected with a C. jejuni wild-type strain. Ectopic expression of CiaI-GFP in epithelial cells yielded a punctate phenotype not observed with the other C. jejuni genes, and this phenotype was abolished by mutation of a dileucine motif located in the carboxy-terminus of the protein. Based on the data, we conclude that CiaI contributes to the ability of C. jejuni to survive within epithelial cells. PMID:21435039

  3. The isolation and characterization of Campylobacter jejuni bacteriophages from free range and indoor poultry.

    PubMed

    Owens, Jane; Barton, Mary D; Heuzenroeder, Michael W

    2013-02-22

    Six hundred and sixty one samples - primarily fresh chicken faeces - were processed to isolate wild type Campylobacter jejuni bacteriophages, via overlay agar methods using C. jejuni NCTC 12662. The aims of this study were to isolate and purify bacteriophages and then test for their ability to lyse field strains of C. jejuni in vitro. Of all samples processed, 130 were positive for bacteriophages. A distinct difference was observed between samples from different poultry enterprises. No bacteriophages could be isolated from indoor broilers. The majority of bacteriophages were isolated from free range poultry - both broilers and egg layers. Bacteriophages were purified and then selected for characterization based on their ability to produce clear lysis on plaque assay, as opposed to turbid plaques. Two hundred and forty one C. jejuni field isolates were tested for sensitivity to the bacteriophages. Lysis was graded subjectively and any minimal lysis was excluded. Using this system, 59.0% of the C. jejuni isolates showed significant sensitivity to at least one bacteriophage. The sensitivity to individual bacteriophages ranged from 10.0% to 32.5% of the C. jejuni isolates. Five bacteriophages were examined by electron microscopy and determined to belong to the Myoviridae family. The physical size, predicted genetic composition and genome size of the bacteriophages correlated well with other reported Campylobacter bacteriophages. The reasons for the observed difference between indoor broilers and free range poultry is unknown, but are postulated to be due to differences in the Campylobacter population in birds under different rearing conditions.

  4. Is allicin able to reduce Campylobacter jejuni colonization in broilers when added to drinking water?

    PubMed

    Robyn, J; Rasschaert, G; Hermans, D; Pasmans, F; Heyndrickx, M

    2013-05-01

    Reducing Campylobacter shedding on the farm could result in a reduction of the number of human campylobacteriosis cases. In this study, we first investigated if allicin, allyl disulfide, and garlic oil extract were able to either prevent C. jejuni growth or kill C. jejuni in vitro. Allyl disulfide and garlic oil extract reduced C. jejuni numbers in vitro below a detectable level at a concentration of 50 mg/kg (no lower concentrations were tested), whereas allicin reduced C. jejuni numbers below a detectable level at a concentration as low as 7.5 mg/kg. In further experiments we screened for the anti-C. jejuni activity of allicin in a fermentation system closely mimicking the broiler cecal environment using cecal microbiota and mucus isolated from C. jejuni-free broilers. During these fermentation experiments, allicin reduced C. jejuni numbers below a detectable level after 24 h at a concentration of 50 mg/kg. In contrast, 25 mg/kg of allicin killed C. jejuni in the first 28 h of incubation, but anti-C. jejuni activity was lost after 48 h of incubation, probably due to the presence of mucin in the growth medium. This had been confirmed in fermentation experiments in the presence of broiler cecal mucus. Based on these results, we performed an in vivo experiment to assess the prevention or reduction of cecal C. jejuni colonization in broiler chickens when allicin was added to drinking water. We demonstrated that allicin in drinking water did not have a statistically significant effect on cecal C. jejuni colonization in broilers. It was assumed, based on in vitro experiments, that the activity of allicin was thwarted by the presence of mucin-containing mucus. Despite promising in vitro results, allicin was not capable of statistically influencing C. jejuni colonization in a broiler flock, although a trend toward lower cecal C. jejuni numbers in allicin-treated broilers was observed.

  5. 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.

  6. Re-sequencing of a virulent strain of Campylobacter jejuni NCTC11168 reveals potential virulence factors.

    PubMed

    Cooper, Kerry K; Cooper, Margarethe A; Zuccolo, Andrea; Joens, Lynn A

    2013-01-01

    In vitro passage of Campylobacter jejuni strains results in phenotypic changes and a general loss of virulence, as is the case with the genome-sequenced strain C. jejuni NCTC11168. Re-sequencing of a virulent strain of NCTC11168 identified 41 SNPs or indels involving 20 genes, four intergenic regions and three pseudogenes. The genes include six motility genes, two chemotaxis genes, three hypothetical genes and a capsule biosynthesis gene, which might have a critical role in C. jejuni virulence. Additionally, we found an insertion in both Cj0676 and Cj1470c, pseudogenes in avirulent NCTC11168, but functional proteins in virulent NCTC11168.

  7. [Research progress in biofilm formation and regulatory mechanism of Campylobacter jejuni].

    PubMed

    Wu, Qingping; Zhong, Xian; Zhang, Jumei

    2016-02-01

    Biofilm of Campylobacter jejuni was formed by cross-linking its extracellular secretion, polysaccharides, various extracellular proteins, nucleic acids etc to enhance its survival in hostile environments, especially for detergents, antibiotics and disinfectants. This paper elaborated C. jejuni biofilm formation and regulation mechanisms in the surface properties of the media, temperatures, gas environment, the regulation of gene etc, also analysed and discussed a variety of biofilm removal practical applications. We hope it can provide a reference for studies on biofilm control of C. jejuni. PMID:27373066

  8. Toll-like receptors recognize distinct proteinase-resistant glycoconjugates in Campylobacter jejuni and Escherichia coli.

    PubMed

    Phongsisay, Vongsavanh; Hara, Hiromitsu; Fujimoto, Shuji

    2015-03-01

    Campylobacter jejuni causes gastroenteritis and autoimmune neuropathy Guillain-Barré syndrome. The mechanism by which C. jejuni infection results in such the hyperimmunity is not completely understood. Host immunity plays an important role in the disease pathogenesis; however, little is known how immune system recognizes this human pathogen. In this study, we report that Toll-like receptors recognize distinct proteinase K-resistant glycoconjugates in C. jejuni and Escherichia coli. Lipopolysaccharide is solely proteinase-resistant glycoconjugate in E. coli. In contrast, C. jejuni possesses at least five different components that are resistant to proteinase digestion and are capable of inducing NF-κB activation through TLR2 and TLR4. Possession of multiple activators of Toll-like receptors may be the unique strategy of C. jejuni to trigger hyperimmunity.

  9. Regulation of oxidative stress resistance in Campylobacter jejuni, a microaerophilic foodborne pathogen

    PubMed Central

    Kim, Jong-Chul; Oh, Euna; Kim, Jinyong; Jeon, Byeonghwa

    2015-01-01

    Campylobacter jejuni is one of the leading bacterial causes of human gastroenteritis. Due to the increasing rates of human campylobacteriosis, C. jejuni is considered as a serious public health concern worldwide. C. jejuni is a microaerophilic, fastidious bacterium. C. jejuni must overcome a wide range of stress conditions during foodborne transmission to humans, such as food preservation and processing conditions, and even in infection of the gastrointestinal tracts of humans. Particularly, this microaerophilic foodborne pathogen must survive in the atmospheric conditions prior to the initiation of infection. C. jejuni possesses unique regulatory mechanisms for oxidative stress resistance. Lacking OxyR and SoxRS that are highly conserved in other Gram-negative foodborne pathogens, C. jejuni modulates the expression of genes involved in oxidative stress resistance mainly via the peroxide resistance regulator and Campylobacter oxidative stress regulator. Based on recent findings of ours and others, in this review, we described how C. jejuni regulates the expression of oxidative stress defense. PMID:26284041

  10. 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

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

    PubMed

    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.

  12. Long-Lasting Outbreak of Erythromycin- and Ciprofloxacin-Resistant Campylobacter jejuni Subspecies jejuni From 2003 to 2013 in Men Who Have Sex With Men, Quebec, Canada.

    PubMed

    Gaudreau, Christiane; Rodrigues-Coutlée, Sophie; Pilon, Pierre A; Coutlée, François; Bekal, Sadjia

    2015-11-15

    From January 2003 to December 2013, sexual transmission of 2 clades of Campylobacter jejuni subspecies jejuni isolates resulted in a prolonged outbreak among men who have sex with men living in Quebec, Canada. The outbreak isolates were acquired locally and were resistant to erythromycin and ciprofloxacin.

  13. Detection and typing of Campylobacter jejuni and Campylobacter coli and analysis of indicator organisms in three waterborne outbreaks in Finland.

    PubMed

    Hänninen, Marja-Liisa; Haajanen, H; Pummi, T; Wermundsen, K; Katila, M-L; Sarkkinen, H; Miettinen, I; Rautelin, H

    2003-03-01

    Waterborne outbreaks associated with contamination of drinking water by Campylobacter jejuni are rather common in the Nordic countries Sweden, Norway, and Finland, where in sparsely populated districts groundwater is commonly used without disinfection. Campylobacters, Escherichia coli, or other coliforms have rarely been detected in potential sources. We studied three waterborne outbreaks in Finland caused by C. jejuni and used sample volumes of 4,000 to 20,000 ml for analysis of campylobacters and sample volumes of 1 to 5,000 ml for analysis of coliforms and E. coli, depending on the sampling site. Multiple samples obtained from possible sources (water distribution systems and environmental water sources) and the use of large sample volumes (several liters) increased the chance of detecting the pathogen C. jejuni in water. Filtration of a large volume (1,000 to 2,000 ml) also increased the rate of detection of coliforms and E. coli. To confirm the association between drinking water contamination and illness, a combination of Penner serotyping and pulsed-field gel electrophoresis (digestion with SmaI and KpnI) was found to be useful. This combination reliably verified similarity or dissimilarity of C. jejuni isolates from patient samples, from drinking water, and from other environmental sources, thus confirming the likely reservoir of an outbreak.

  14. Antimicrobial Susceptibility Profiles of Human Campylobacter jejuni Isolates and Association with Phylogenetic Lineages

    PubMed Central

    Cha, Wonhee; Mosci, Rebekah; Wengert, Samantha L.; Singh, Pallavi; Newton, Duane W.; Salimnia, Hossein; Lephart, Paul; Khalife, Walid; Mansfield, Linda S.; Rudrik, James T.; Manning, Shannon D.

    2016-01-01

    Campylobacter jejuni is a zoonotic pathogen and the most common bacterial cause of human gastroenteritis worldwide. With the increase of antibiotic resistance to fluoroquinolones and macrolides, the drugs of choice for treatment, C. jejuni was recently classified as a serious antimicrobial resistant threat. Here, we characterized 94 C. jejuni isolates collected from patients at four Michigan hospitals in 2011 and 2012 to determine the frequency of resistance and association with phylogenetic lineages. The prevalence of resistance to fluoroquinolones (19.1%) and macrolides (2.1%) in this subset of C. jejuni isolates from Michigan was similar to national reports. High frequencies of fluoroquinolone-resistant C. jejuni isolates, however, were recovered from patients with a history of foreign travel. A high proportion of these resistant isolates were classified as multilocus sequence type (ST)-464, a fluoroquinolone-resistant lineage that recently emerged in Europe. A significantly higher prevalence of tetracycline-resistant C. jejuni was also found in Michigan and resistant isolates were more likely to represent ST-982, which has been previously recovered from ruminants and the environment in the U.S. Notably, patients with tetracycline-resistant C. jejuni infections were more likely to have contact with cattle. These outcomes prompt the need to monitor the dissemination and diversification of imported fluoroquinolone-resistant C. jejuni strains and to investigate the molecular epidemiology of C. jejuni recovered from cattle and farm environments to guide mitigation strategies. PMID:27199922

  15. Virulence and Genomic Feature of Multidrug Resistant Campylobacter jejuni Isolated from Broiler Chicken

    PubMed Central

    Hao, Haihong; Ren, Ni; Han, Jing; Foley, Steven L.; Iqbal, Zahid; Cheng, Guyue; Kuang, Xiuhua; Liu, Jie; Liu, Zhenli; Dai, Menghong; Wang, Yulian; Yuan, Zonghui

    2016-01-01

    The aim of this study was to reveal the molecular mechanism involved in multidrug resistance and virulence of Campylobacter jejuni isolated from broiler chickens. The virulence of six multidrug resistant C. jejuni was determined by in vitro and in vivo methods. The de novo whole genome sequencing technology and molecular biology methods were used to analyze the genomic features associated with the multidrug resistance and virulence of a selected isolate (C. jejuni 1655). The comparative genomic analyses revealed a large number of single nucleotide polymorphisms, deletions, rearrangements, and inversions in C. jejuni 1655 compared to reference C. jejuni genomes. The co-emergence of Thr-86-Ile mutation in gyrA gene, A2075G mutation in 23S rRNA gene, tetO, aphA and aadE genes and pTet plasmid in C. jejuni 1655 contributed its multidrug resistance to fluoroquinolones, macrolides, tetracycline, and aminoglycosides. The combination of multiple virulence genes may work together to confer the relative higher virulence in C. jejuni 1655. The co-existence of mobile gene elements (e.g., pTet) and CRISPR-Cas system in C. jejuni 1655 may play an important role in the gene transfer and immune defense. The present study provides basic information of phenotypic and genomic features of C. jejuni 1655, a strain recently isolated from a chicken displaying multidrug resistance and relatively high level of virulence. PMID:27790202

  16. Antimicrobial Susceptibility Profiles of Human Campylobacter jejuni Isolates and Association with Phylogenetic Lineages.

    PubMed

    Cha, Wonhee; Mosci, Rebekah; Wengert, Samantha L; Singh, Pallavi; Newton, Duane W; Salimnia, Hossein; Lephart, Paul; Khalife, Walid; Mansfield, Linda S; Rudrik, James T; Manning, Shannon D

    2016-01-01

    Campylobacter jejuni is a zoonotic pathogen and the most common bacterial cause of human gastroenteritis worldwide. With the increase of antibiotic resistance to fluoroquinolones and macrolides, the drugs of choice for treatment, C. jejuni was recently classified as a serious antimicrobial resistant threat. Here, we characterized 94 C. jejuni isolates collected from patients at four Michigan hospitals in 2011 and 2012 to determine the frequency of resistance and association with phylogenetic lineages. The prevalence of resistance to fluoroquinolones (19.1%) and macrolides (2.1%) in this subset of C. jejuni isolates from Michigan was similar to national reports. High frequencies of fluoroquinolone-resistant C. jejuni isolates, however, were recovered from patients with a history of foreign travel. A high proportion of these resistant isolates were classified as multilocus sequence type (ST)-464, a fluoroquinolone-resistant lineage that recently emerged in Europe. A significantly higher prevalence of tetracycline-resistant C. jejuni was also found in Michigan and resistant isolates were more likely to represent ST-982, which has been previously recovered from ruminants and the environment in the U.S. Notably, patients with tetracycline-resistant C. jejuni infections were more likely to have contact with cattle. These outcomes prompt the need to monitor the dissemination and diversification of imported fluoroquinolone-resistant C. jejuni strains and to investigate the molecular epidemiology of C. jejuni recovered from cattle and farm environments to guide mitigation strategies. PMID:27199922

  17. Prevalence in bulk tank milk and epidemiology of Campylobacter jejuni in dairy herds in Northern Italy.

    PubMed

    Bianchini, Valentina; Borella, Laura; Benedetti, Valentina; Parisi, Antonio; Miccolupo, Angela; Santoro, Eliana; Recordati, Camilla; Luini, Mario

    2014-03-01

    Thermotolerant Campylobacter spp. are frequently the cause of human gastroenteritis and have assumed more importance in Italy following the increased consumption of raw milk. Our objectives were to determine the prevalence and genotypes of Campylobacter spp. in dairy herds and to investigate the possible sources of bulk milk contamination. Bulk milk from dairy herds (n = 282) was cultured for Campylobacter spp. and Enterobacteriaceae. At three Campylobacter jejuni-positive farms, bovine feces, pigeon intestines, milk, and water points were also investigated. Isolates were identified by PCR and genotyped using multilocus sequence typing (MLST). C. jejuni was detected in 34 (12%) bulk milk samples. The strains belonged to 14 sequence types, and the most common clonal complexes were CC-21, CC-48, and CC-403. No association was demonstrated between the presence of C. jejuni and high levels of Enterobacteriaceae in bulk milk. At the three farms examined, C. jejuni was isolated from bovine feces (25/82 [30.5%]), pigeon intestines (13/60 [21.7%]), bulk milk (10/24 [41.7%]), and water points (4/16 [25%]). MLST revealed lineages that were common between milk and bovine feces but distinct between cattle and pigeons. In one herd, C. jejuni with the same genotype was isolated repeatedly from bulk milk and a cow with an udder infection. Our results showed a high prevalence of C. jejuni in bulk milk and suggested that udder excretion, in addition to fecal matter, may be a route of bulk milk contamination. MLST analysis indicated that pigeons are probably not relevant for the transmission of C. jejuni to cattle and for milk contamination.

  18. Prevalence in Bulk Tank Milk and Epidemiology of Campylobacter jejuni in Dairy Herds in Northern Italy

    PubMed Central

    Bianchini, Valentina; Borella, Laura; Benedetti, Valentina; Parisi, Antonio; Miccolupo, Angela; Santoro, Eliana; Recordati, Camilla

    2014-01-01

    Thermotolerant Campylobacter spp. are frequently the cause of human gastroenteritis and have assumed more importance in Italy following the increased consumption of raw milk. Our objectives were to determine the prevalence and genotypes of Campylobacter spp. in dairy herds and to investigate the possible sources of bulk milk contamination. Bulk milk from dairy herds (n = 282) was cultured for Campylobacter spp. and Enterobacteriaceae. At three Campylobacter jejuni-positive farms, bovine feces, pigeon intestines, milk, and water points were also investigated. Isolates were identified by PCR and genotyped using multilocus sequence typing (MLST). C. jejuni was detected in 34 (12%) bulk milk samples. The strains belonged to 14 sequence types, and the most common clonal complexes were CC-21, CC-48, and CC-403. No association was demonstrated between the presence of C. jejuni and high levels of Enterobacteriaceae in bulk milk. At the three farms examined, C. jejuni was isolated from bovine feces (25/82 [30.5%]), pigeon intestines (13/60 [21.7%]), bulk milk (10/24 [41.7%]), and water points (4/16 [25%]). MLST revealed lineages that were common between milk and bovine feces but distinct between cattle and pigeons. In one herd, C. jejuni with the same genotype was isolated repeatedly from bulk milk and a cow with an udder infection. Our results showed a high prevalence of C. jejuni in bulk milk and suggested that udder excretion, in addition to fecal matter, may be a route of bulk milk contamination. MLST analysis indicated that pigeons are probably not relevant for the transmission of C. jejuni to cattle and for milk contamination. PMID:24413598

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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...

  20. Complete genome sequence of Campylobacter jejuni RM1285 a rod-shaped morphological variant

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Campylobacter jejuni is a spiral-shaped Gram-negative food-borne human pathogen found on poultry products. Strain RM1285 is a rod-shaped variant of this species. The genome of RM1285 was determined to be 1,635,803 bp with a G+C content of 30.5%....

  1. Population Diversity of Campylobacter jejuni in Poultry and Its Dynamic of Contamination in Chicken Meat.

    PubMed

    Marotta, Francesca; Garofolo, Giuliano; Di Donato, Guido; Aprea, Giuseppe; Platone, Ilenia; Cianciavicchia, Silvia; Alessiani, Alessandra; Di Giannatale, Elisabetta

    2015-01-01

    This study aimed to analyse the diversity of the Campylobacter jejuni population in broilers and to evaluate the major source of contamination in poultry meat. Eight rearing cycles over one year provided samples from three different broiler farms processed at the same slaughterhouse. A total of 707 C. jejuni were isolated from cloacal swabs before slaughter and from the breast skin of carcasses after slaughter and after chilling. All suspected Campylobacter colonies were identified with PCR assays and C. jejuni was genotyped by sequence analysis of the flaA short variable region (SVR) and by pulsed-field gel electrophoresis (PFGE) using SmaI enzyme. Phenotypic antibiotic resistance profiles were also assayed using minimal inhibitory concentration (MIC). The flocks carried many major C. jejuni clones possibly carrying over the rearing cycles, but cross contamination between farms may happen. Many isolates were resistant to fluoroquinolones, raising an issue of high public concern. Specific Campylobacter populations could be harboured within each poultry farm, with the ability to contaminate chickens during each new cycle. Thus, although biosecurity measures are applied, with a persistent source of contamination, they cannot be efficient. The role of the environment needs further investigation to better address strategies to control Campylobacter.

  2. Improved isolation protocol to detect high molecular weight polysaccharide structures of Campylobacter jejuni.

    PubMed

    Kovács, Judit K; Felső, Péter; Emődy, Levente; Schneider, György; Kocsis, Béla

    2014-12-01

    Simple detection of high molecular weight, LPS-like structures of Campylobacter jejuni is still an unsolved problem. A phenol-free extraction method for the detection of HMW polysaccharide was developed without the need for Western blot. This method provides a reliable technique for large-scale screening and comparative characterization study of different isolates.

  3. EFFECTS OF UNIQUE COMMUNITIES OF INTESTINAL MICROBIOTA ON CAMPYLOBACTER JEJUNI COLONIZATION

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Campylobacter jejuni has been identified as a major source of food-borne illness worldwide, with an annual estimate of 2 million cases in the United States alone. Most common source of infection is through consumption of poultry, whose animal populations are nearly ubiquitously contaminated with C....

  4. Population Diversity of Campylobacter jejuni in Poultry and Its Dynamic of Contamination in Chicken Meat

    PubMed Central

    Marotta, Francesca; Garofolo, Giuliano; Di Donato, Guido; Aprea, Giuseppe; Platone, Ilenia; Cianciavicchia, Silvia; Alessiani, Alessandra; Di Giannatale, Elisabetta

    2015-01-01

    This study aimed to analyse the diversity of the Campylobacter jejuni population in broilers and to evaluate the major source of contamination in poultry meat. Eight rearing cycles over one year provided samples from three different broiler farms processed at the same slaughterhouse. A total of 707  C. jejuni were isolated from cloacal swabs before slaughter and from the breast skin of carcasses after slaughter and after chilling. All suspected Campylobacter colonies were identified with PCR assays and C. jejuni was genotyped by sequence analysis of the flaA short variable region (SVR) and by pulsed-field gel electrophoresis (PFGE) using SmaI enzyme. Phenotypic antibiotic resistance profiles were also assayed using minimal inhibitory concentration (MIC). The flocks carried many major C. jejuni clones possibly carrying over the rearing cycles, but cross contamination between farms may happen. Many isolates were resistant to fluoroquinolones, raising an issue of high public concern. Specific Campylobacter populations could be harboured within each poultry farm, with the ability to contaminate chickens during each new cycle. Thus, although biosecurity measures are applied, with a persistent source of contamination, they cannot be efficient. The role of the environment needs further investigation to better address strategies to control Campylobacter. PMID:26543870

  5. Beyond gangliosides: Multiple forms of glycan mimicry exhibited by Campylobacter jejuni in its lipooligosaccharide (LOS)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Campylobacter jejuni is well known for synthesizing ganglioside mimics within the glycan component of its lipooligosaccharide (LOS), which have been implicated in triggering Guillain-Barré syndrome (GBS). We now confirm that this pathogen is capable of synthesizing a much broader spectrum of host g...

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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 ...

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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...

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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...

  9. Draft genome sequences of two Campylobacter jejuni clinical isolates, NW and D2600.

    PubMed

    Jerome, John P; Klahn, Brian D; Bell, Julia A; Barrick, Jeffrey E; Brown, C Titus; Mansfield, Linda S

    2012-10-01

    The Campylobacter jejuni human clinical isolates NW and D2600 colonized C57BL/6 interleukin-10-deficient (IL-10(-/-)) mice without inducing a robust inflammatory response (J. A. Bell et al., BMC Microbiol. 9:57, 2009). We announce draft genome sequences of NW and D2600 to facilitate comparisons with strains that induce gastrointestinal inflammation in this mouse model.

  10. Efficacy of media and methods for detecting and enumerating Campylobacter jejuni in refrigerated chicken meat.

    PubMed Central

    Beuchat, L R

    1985-01-01

    A study was undertaken to compare several enrichment and direct isolation media for their suitability to detect and enumerate five strains of Campylobacter jejuni in refrigerated (5 degrees C) chicken meat. The influence of CO2 on survival at 5 degrees C was also investigated. Selective enrichment media evaluated included Preston broth (PB), selective semisolid brucella medium (SSBM), Campylobacter enrichment broth (CEB), VTP brucella-FBP broth (VTP), Rosef and Kapperud Campylobacter enrichment broth (RKCEB), and Doyle and Roman enrichment broth (DREB). Direct isolation agars included Campy brucella agar (CBAP), blood-free Campylobacter medium (BFCM) and modified Butzler agar (MBA). Comminuted chicken meat was inoculated with C. jejuni, sealed under atmospheric gas or CO2, and stored at 5 degrees C for up to 21 days. Viable population was determined by the most-probable-number technique (PB, SSBM, CEB, VTP, and RKCEB, followed by plating on CBAP, BFCM, and MBA), enrichment on DREB, followed by plating on CBAP, BFCM, and MBA, and direct isolation on CBAP, BFCM, and MBA. Without exception, direct plating of samples was superior to the most-probable-number technique for enumerating C. jejuni; MBA was inferior to CBAP and BFCM, and DREB performed at least as well as other enrichment media evaluated. Carbon dioxide afforded protection against death of three of the five strains of C. jejuni tested. PMID:4083888

  11. 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.

  12. L-fucose influences chemotaxis and biofilm formation in Campylobacter jejuni.

    PubMed

    Dwivedi, Ritika; Nothaft, Harald; Garber, Jolene; Xin Kin, Lin; Stahl, Martin; Flint, Annika; van Vliet, Arnoud H M; Stintzi, Alain; Szymanski, Christine M

    2016-08-01

    Campylobacter jejuni and Campylobacter coli are zoonotic pathogens once considered asaccharolytic, but are now known to encode pathways for glucose and fucose uptake/metabolism. For C. jejuni, strains with the fuc locus possess a competitive advantage in animal colonization models. We demonstrate that this locus is present in > 50% of genome-sequenced strains and is prevalent in livestock-associated isolates of both species. To better understand how these campylobacters sense nutrient availability, we examined biofilm formation and chemotaxis to fucose. C. jejuni NCTC11168 forms less biofilms in the presence of fucose, although its fucose permease mutant (fucP) shows no change. In a newly developed chemotaxis assay, both wild-type and the fucP mutant are chemotactic towards fucose. C. jejuni 81-176 naturally lacks the fuc locus and is unable to swim towards fucose. Transfer of the NCTC11168 locus into 81-176 activated fucose uptake and chemotaxis. Fucose chemotaxis also correlated with possession of the pathway for C. jejuni RM1221 (fuc+) and 81116 (fuc-). Systematic mutation of the NCTC11168 locus revealed that Cj0485 is necessary for fucose metabolism and chemotaxis. This study suggests that components for fucose chemotaxis are encoded within the fuc locus, but downstream signals only in fuc + strains, are involved in coordinating fucose availability with biofilm development. PMID:27145048

  13. 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

  14. L-fucose influences chemotaxis and biofilm formation in Campylobacter jejuni.

    PubMed

    Dwivedi, Ritika; Nothaft, Harald; Garber, Jolene; Xin Kin, Lin; Stahl, Martin; Flint, Annika; van Vliet, Arnoud H M; Stintzi, Alain; Szymanski, Christine M

    2016-08-01

    Campylobacter jejuni and Campylobacter coli are zoonotic pathogens once considered asaccharolytic, but are now known to encode pathways for glucose and fucose uptake/metabolism. For C. jejuni, strains with the fuc locus possess a competitive advantage in animal colonization models. We demonstrate that this locus is present in > 50% of genome-sequenced strains and is prevalent in livestock-associated isolates of both species. To better understand how these campylobacters sense nutrient availability, we examined biofilm formation and chemotaxis to fucose. C. jejuni NCTC11168 forms less biofilms in the presence of fucose, although its fucose permease mutant (fucP) shows no change. In a newly developed chemotaxis assay, both wild-type and the fucP mutant are chemotactic towards fucose. C. jejuni 81-176 naturally lacks the fuc locus and is unable to swim towards fucose. Transfer of the NCTC11168 locus into 81-176 activated fucose uptake and chemotaxis. Fucose chemotaxis also correlated with possession of the pathway for C. jejuni RM1221 (fuc+) and 81116 (fuc-). Systematic mutation of the NCTC11168 locus revealed that Cj0485 is necessary for fucose metabolism and chemotaxis. This study suggests that components for fucose chemotaxis are encoded within the fuc locus, but downstream signals only in fuc + strains, are involved in coordinating fucose availability with biofilm development.

  15. 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. PMID:26735032

  16. Rapid Detection of Campylobacter jejuni, Campylobacter coli, and Campylobacter lari in Fresh Chicken Meat and By-Products in Bangkok, Thailand, Using Modified Multiplex PCR.

    PubMed

    Saiyudthong, S; Phusri, K; Buates, S

    2015-07-01

    A multiplex PCR assay for simultaneous detection and differentiation of Campylobacter jejuni, Campylobacter coli, and Campylobacter lari was developed and validated to assess the occurrence of these bacteria in fresh chicken meat and by-products in Bangkok, Thailand, by using a new combination of four previously published PCR primers for C. jejuni, C. coli, C. lari, and a universal 16S rDNA gene as an internal control. The specificity was determined by using 13 strains of other bacteria. With pure culture DNA, the detection limit was 0.017 ng/PCR for C. jejuni and C. coli and was 0.016 ng/PCR for C. lari. It can detect 10 CFU of C. jejuni, C. coli, and C. lari in 2 g of chicken meat within a 16-h enrichment time. Our multiplex PCR assay was applied for identification of Campylobacter spp. in 122 supermarket samples and 108 fresh market samples. Of the 230 samples evaluated by multiplex PCR, 54.0, 3.3, and 10.7% of supermarket samples were positive for C. jejuni, C. coli, and mixed C. jejuni and C. coli, respectively, and 56.5 and 33.3% of fresh market samples were positive for C. jejuni and mixed C. jejuni and C. coli, respectively. No sample was positive for C. lari. Fresh market samples had significantly higher C. jejuni and C. coli contamination than those from supermarkets (relative risk: 1.3; P = 0.0001). Compared with the culture method (a gold standard), the sensitivity, specificity, positive predictive value, negative predictive value, and diagnostic accuracy of multiplex PCR were 97.7, 86.8, 96.1, 92.0, and 95.2%, respectively. No significant difference was observed between results from two methods (P = 0.55). Therefore, the established multiplex PCR was not only rapid and easy to perform but had a high sensitivity and specificity to distinguish between C. jejuni, C. coli, and C. lari, even in samples containing mixed contamination. Our study indicated that fresh chicken meat and by-products from fresh markets were significantly less hygienic than those

  17. 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

  18. Packaging of Campylobacter jejuni into Multilamellar Bodies by the Ciliate Tetrahymena pyriformis

    PubMed Central

    Trigui, Hana; Paquet, Valérie E.; Charette, Steve J.

    2016-01-01

    Campylobacter jejuni is the leading cause of bacterial gastroenteritis worldwide. Transmission to humans occurs through consumption of contaminated food or water. The conditions affecting the persistence of C. jejuni in the environment are poorly understood. Some protozoa package and excrete bacteria into multilamellar bodies (MLBs). Packaged bacteria are protected from deleterious conditions, which increases their survival. We hypothesized that C. jejuni could be packaged under aerobic conditions by the amoeba Acanthamoeba castellanii or the ciliate Tetrahymena pyriformis, both of which are able to package other pathogenic bacteria. A. castellanii did not produce MLBs containing C. jejuni. In contrast, when incubated with T. pyriformis, C. jejuni was ingested, packaged in MLBs, and then expelled into the milieu. The viability of the bacteria inside MLBs was confirmed by microscopic analyses. The kinetics of C. jejuni culturability showed that packaging increased the survival of C. jejuni up to 60 h, in contrast to the strong survival defect seen in ciliate-free culture. This study suggests that T. pyriformis may increase the risk of persistence of C. jejuni in the environment and its possible transmission between different reservoirs in food and potable water through packaging. PMID:26921427

  19. Polyphosphate and associated enzymes as global regulators of stress response and virulence in Campylobacter jejuni

    PubMed Central

    Kumar, Anand; Gangaiah, Dharanesh; Torrelles, Jordi B; Rajashekara, Gireesh

    2016-01-01

    Campylobacter jejuni (C. jejuni), a Gram-negative microaerophilic bacterium, is a predominant cause of bacterial foodborne gastroenteritis in humans worldwide. Despite its importance as a major foodborne pathogen, our understanding of the molecular mechanisms underlying C. jejuni stress survival and pathogenesis is limited. Inorganic polyphosphate (poly P) has been shown to play significant roles in bacterial resistance to stress and virulence in many pathogenic bacteria. C. jejuni contains the complete repertoire of enzymes required for poly P metabolism. Recent work in our laboratory and others have demonstrated that poly P controls a plethora of C. jejuni properties that impact its ability to survive in the environment as well as to colonize/infect mammalian hosts. This review article summarizes the current literature on the role of poly P in C. jejuni stress survival and virulence and discusses on how poly P-related enzymes can be exploited for therapeutic/prevention purposes. Additionally, the review article identifies potential areas for future investigation that would enhance our understanding of the role of poly P in C. jejuni and other bacteria, which ultimately would facilitate design of effective therapeutic/preventive strategies to reduce not only the burden of C. jejuni-caused foodborne infections but also of other bacterial infections in humans. PMID:27672264

  20. Polyphosphate and associated enzymes as global regulators of stress response and virulence in Campylobacter jejuni.

    PubMed

    Kumar, Anand; Gangaiah, Dharanesh; Torrelles, Jordi B; Rajashekara, Gireesh

    2016-09-01

    Campylobacter jejuni (C. jejuni), a Gram-negative microaerophilic bacterium, is a predominant cause of bacterial foodborne gastroenteritis in humans worldwide. Despite its importance as a major foodborne pathogen, our understanding of the molecular mechanisms underlying C. jejuni stress survival and pathogenesis is limited. Inorganic polyphosphate (poly P) has been shown to play significant roles in bacterial resistance to stress and virulence in many pathogenic bacteria. C. jejuni contains the complete repertoire of enzymes required for poly P metabolism. Recent work in our laboratory and others have demonstrated that poly P controls a plethora of C. jejuni properties that impact its ability to survive in the environment as well as to colonize/infect mammalian hosts. This review article summarizes the current literature on the role of poly P in C. jejuni stress survival and virulence and discusses on how poly P-related enzymes can be exploited for therapeutic/prevention purposes. Additionally, the review article identifies potential areas for future investigation that would enhance our understanding of the role of poly P in C. jejuni and other bacteria, which ultimately would facilitate design of effective therapeutic/preventive strategies to reduce not only the burden of C. jejuni-caused foodborne infections but also of other bacterial infections in humans. PMID:27672264

  1. Evidence of broiler meat contamination with post-disinfection strains of Campylobacter jejuni from slaughterhouse.

    PubMed

    Kudirkienė, Eglė; Bunevičienė, Jurgita; Brøndsted, Lone; Ingmer, Hanne; Olsen, John Elmerdahl; Malakauskas, Mindaugas

    2011-03-01

    While cross-contamination from equipment and scalding water containing Campylobacter jejuni is considered the main route of broiler carcass contamination during slaughtering, alternative sources of C. jejuni may have been overlooked because only a limited number of studies focus on sampling of one broiler flock along the entire food chain and not many include the slaughterhouse environment. In the present study we have traced the changes of C. jejuni genotypes within one broiler flock from the beginning of rearing to the final product at the slaughterhouse with the aim to evaluate the dynamics and possible sources of carcass contamination with C. jejuni. Genotyping of 345 isolates of C. jejuni by flaA-RFLP revealed ten different flaA genotypes of C. jejuni along the broiler meat production chain. Broiler fillets were mainly contaminated with flaA genotypes found on the surfaces of slaughterhouse equipment and in the scalding water after cleaning and disinfection. Finally, it was clearly demonstrated that C. jejuni isolates remaining in the slaughterhouse environment after disinfection is a potential source of broiler meat contamination. Thus, identification of the mechanisms that allow such strains to persist in the slaughterhouse and survive cleaning is important for the establishment of future practices that will ensure sufficient reduction of C. jejuni in the slaughterhouse environment.

  2. Analysis of the Campylobacter jejuni Genome by SMRT DNA Sequencing Identifies Restriction-Modification Motifs

    PubMed Central

    O’Loughlin, Jason L.; Eucker, Tyson P.; Chavez, Juan D.; Samuelson, Derrick R.; Neal-McKinney, Jason; Gourley, Christopher R.; Bruce, James E.; Konkel, Michael E.

    2015-01-01

    Campylobacter jejuni is a leading bacterial cause of human gastroenteritis. The goal of this study was to analyze the C. jejuni F38011 strain, recovered from an individual with severe enteritis, at a genomic and proteomic level to gain insight into microbial processes. The C. jejuni F38011 genome is comprised of 1,691,939 bp, with a mol.% (G+C) content of 30.5%. PacBio sequencing coupled with REBASE analysis was used to predict C. jejuni F38011 genomic sites and enzymes that may be involved in DNA restriction-modification. A total of five putative methylation motifs were identified as well as the C. jejuni enzymes that could be responsible for the modifications. Peptides corresponding to the deduced amino acid sequence of the C. jejuni enzymes were identified using proteomics. This work sets the stage for studies to dissect the precise functions of the C. jejuni putative restriction-modification enzymes. Taken together, the data generated in this study contributes to our knowledge of the genomic content, methylation profile, and encoding capacity of C. jejuni. PMID:25695747

  3. Polyphosphate and associated enzymes as global regulators of stress response and virulence in Campylobacter jejuni

    PubMed Central

    Kumar, Anand; Gangaiah, Dharanesh; Torrelles, Jordi B; Rajashekara, Gireesh

    2016-01-01

    Campylobacter jejuni (C. jejuni), a Gram-negative microaerophilic bacterium, is a predominant cause of bacterial foodborne gastroenteritis in humans worldwide. Despite its importance as a major foodborne pathogen, our understanding of the molecular mechanisms underlying C. jejuni stress survival and pathogenesis is limited. Inorganic polyphosphate (poly P) has been shown to play significant roles in bacterial resistance to stress and virulence in many pathogenic bacteria. C. jejuni contains the complete repertoire of enzymes required for poly P metabolism. Recent work in our laboratory and others have demonstrated that poly P controls a plethora of C. jejuni properties that impact its ability to survive in the environment as well as to colonize/infect mammalian hosts. This review article summarizes the current literature on the role of poly P in C. jejuni stress survival and virulence and discusses on how poly P-related enzymes can be exploited for therapeutic/prevention purposes. Additionally, the review article identifies potential areas for future investigation that would enhance our understanding of the role of poly P in C. jejuni and other bacteria, which ultimately would facilitate design of effective therapeutic/preventive strategies to reduce not only the burden of C. jejuni-caused foodborne infections but also of other bacterial infections in humans.

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

    PubMed

    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

  5. Passive immunization to reduce Campylobacter jejuni colonization and transmission in broiler chickens.

    PubMed

    Hermans, David; Van Steendam, Katleen; Verbrugghe, Elin; Verlinden, Marc; Martel, An; Seliwiorstow, Tomasz; Heyndrickx, Marc; Haesebrouck, Freddy; De Zutter, Lieven; Deforce, Dieter; Pasmans, Frank

    2014-03-04

    Campylobacter jejuni is the most common cause of bacterium-mediated diarrheal disease in humans worldwide. Poultry products are considered the most important source of C. jejuni infections in humans but to date no effective strategy exists to eradicate this zoonotic pathogen from poultry production. Here, the potential use of passive immunization to reduce Campylobacter colonization in broiler chicks was examined. For this purpose, laying hens were immunized with either a whole-cell lysate or the hydrophobic protein fraction of C. jejuni and their eggs were collected. In vitro tests validated the induction of specific ImmunoglobulinY (IgY) against C. jejuni in the immunized hens' egg yolks, in particular. In seeder experiments, preventive administration of hyperimmune egg yolk significantly (P < 0.01) reduced bacterial counts of seeder animals three days after oral inoculation with approximately 104 cfu C. jejuni, compared with control birds. Moreover, transmission to non-seeder birds was dramatically reduced (hydrophobic protein fraction) or even completely prevented (whole-cell lysate). Purified IgY promoted bacterial binding to chicken intestinal mucus, suggesting enhanced mucosal clearance in vivo. Western blot analysis in combination with mass spectrometry after two-dimensional gel-electrophoresis revealed immunodominant antigens of C. jejuni that are involved in a variety of cell functions, including chemotaxis and adhesion. Some of these (AtpA, EF-Tu, GroEL and CtpA) are highly conserved proteins and could be promising targets for the development of subunit vaccines.

  6. Engineering the Campylobacter jejuni N-glycan to create an effective chicken vaccine

    PubMed Central

    Nothaft, Harald; Davis, Brandi; Lock, Yee Ying; Perez-Munoz, Maria Elisa; Vinogradov, Evgeny; Walter, Jens; Coros, Colin; Szymanski, Christine M.

    2016-01-01

    Campylobacter jejuni is a predominant cause of human gastroenteritis worldwide. Source-attribution studies indicate that chickens are the main reservoir for infection, thus elimination of C. jejuni from poultry would significantly reduce the burden of human disease. We constructed glycoconjugate vaccines combining the conserved C. jejuni N-glycan with a protein carrier, GlycoTag, or fused to the Escherichia coli lipopolysaccharide-core. Vaccination of chickens with the protein-based or E. coli-displayed glycoconjugate showed up to 10-log reduction in C. jejuni colonization and induced N-glycan-specific IgY responses. Moreover, the live E. coli vaccine was cleared prior to C. jejuni challenge and no selection for resistant campylobacter variants was observed. Analyses of the chicken gut communities revealed that the live vaccine did not alter the composition or complexity of the microbiome, thus representing an effective and low-cost strategy to reduce C. jejuni in chickens and its subsequent entry into the food chain. PMID:27221144

  7. Detection of Campylobacter jejuni in food and poultry viscera using immunomagnetic separation and microtitre hybridization.

    PubMed

    Lamoureux, M; MacKay, A; Messier, S; Fliss, I; Blais, B W; Holley, R A; Simard, R E

    1997-11-01

    Thermophillic Campylobacter and Camp. jejuni were detected from samples of chicken liver, gall bladder, muscle and contaminated milk and chicken meat after an enrichment step by using immunomagnetic capture of cells with monoclonal antibody against a specific outer membrane protein of thermophilic Campylobacter. The detection of captured cells was achieved using two different hybridization methods. In one of the methods, the captured cells were lysed by guanidine isothiocyanate and the 23S rRNA was reacted with a microtitre plate-immobilized rDNA probe specific for thermophilic Campylobacter. In the other method, the captured cells were subjected to lysis by ultrasonication and the genomic DNA reacted with a microtitre plate-immobilized RNA probe specific for Camp.jejuni. Detection of the RNA-DNA hybrids formed in the wells was carried out using a monoclonal anti-RNA-DNA hybrid antibody.

  8. FdhTU-Modulated Formate Dehydrogenase Expression and Electron Donor Availability Enhance Recovery of Campylobacter jejuni following Host Cell Infection

    PubMed Central

    Pryjma, Mark; Apel, Dmitry; Huynh, Steven; Parker, Craig T.

    2012-01-01

    Campylobacter jejuni is a food-borne bacterial pathogen that colonizes the intestinal tract and causes severe gastroenteritis. Interaction with host epithelial cells is thought to enhance severity of disease, and the ability of C. jejuni to modulate its metabolism in different in vivo and environmental niches contributes to its success as a pathogen. A C. jejuni operon comprising two genes that we designated fdhT (CJJ81176_1492) and fdhU (CJJ81176_1493) is conserved in many bacterial species. Deletion of fdhT or fdhU in C. jejuni resulted in apparent defects in adherence and/or invasion of Caco-2 epithelial cells when assessed by CFU enumeration on standard Mueller-Hinton agar. However, fluorescence microscopy indicated that each mutant invaded cells at wild-type levels, instead suggesting roles for FdhTU in either intracellular survival or postinvasion recovery. The loss of fdhU caused reduced mRNA levels of formate dehydrogenase (FDH) genes and a severe defect in FDH activity. Cell infection phenotypes of a mutant deleted for the FdhA subunit of FDH and an ΔfdhU ΔfdhA double mutant were similar to those of a ΔfdhU mutant, which likewise suggested that FdhU and FdhA function in the same pathway. Cell infection assays followed by CFU enumeration on plates supplemented with sodium sulfite abolished the ΔfdhU and ΔfdhA mutant defects and resulted in significantly enhanced recovery of all strains, including wild type, at the invasion and intracellular survival time points. Collectively, our data indicate that FdhTU and FDH are required for optimal recovery following cell infection and suggest that C. jejuni alters its metabolic potential in the intracellular environment. PMID:22636777

  9. Adhesion, Biofilm Formation, and Genomic Features of Campylobacter jejuni Bf, an Atypical Strain Able to Grow under Aerobic Conditions.

    PubMed

    Bronnec, Vicky; Turoňová, Hana; Bouju, Agnès; Cruveiller, Stéphane; Rodrigues, Ramila; Demnerova, Katerina; Tresse, Odile; Haddad, Nabila; Zagorec, Monique

    2016-01-01

    Campylobacter jejuni is the leading cause of bacterial enteritis in Europe. Human campylobacteriosis cases are frequently associated to the consumption of contaminated poultry meat. To survive under environmental conditions encountered along the food chain, i.e., from poultry digestive tract its natural reservoir to the consumer's plate, this pathogen has developed adaptation mechanisms. Among those, biofilm lifestyle has been suggested as a strategy to survive in the food environment and under atmospheric conditions. Recently, the clinical isolate C. jejuni Bf has been shown to survive and grow under aerobic conditions, a property that may help this strain to better survive along the food chain. The aim of this study was to evaluate the adhesion capacity of C. jejuni Bf and its ability to develop a biofilm. C. jejuni Bf can adhere to abiotic surfaces and to human epithelial cells, and can develop biofilm under both microaerobiosis and aerobiosis. These two conditions have no influence on this strain, unlike results obtained with the reference strain C. jejuni 81-176, which harbors only planktonic cells under aerobic conditions. Compared to 81-176, the biofilm of C. jejuni Bf is more homogenous and cell motility at the bottom of biofilm was not modified whatever the atmosphere used. C. jejuni Bf whole genome sequence did not reveal any gene unique to this strain, suggesting that its unusual property does not result from acquisition of new genetic material. Nevertheless some genetic particularities seem to be shared only between Bf and few others strains. Among the main features of C. jejuni Bf genome we noticed (i) a complete type VI secretion system important in pathogenicity and environmental adaptation; (ii) a mutation in the oorD gene involved in oxygen metabolism; and (iii) the presence of an uncommon insertion of a 72 amino acid coding sequence upstream from dnaK, which is involved in stress resistance. Therefore, the atypical behavior of this strain under

  10. Adhesion, Biofilm Formation, and Genomic Features of Campylobacter jejuni Bf, an Atypical Strain Able to Grow under Aerobic Conditions

    PubMed Central

    Bronnec, Vicky; Turoňová, Hana; Bouju, Agnès; Cruveiller, Stéphane; Rodrigues, Ramila; Demnerova, Katerina; Tresse, Odile; Haddad, Nabila; Zagorec, Monique

    2016-01-01

    Campylobacter jejuni is the leading cause of bacterial enteritis in Europe. Human campylobacteriosis cases are frequently associated to the consumption of contaminated poultry meat. To survive under environmental conditions encountered along the food chain, i.e., from poultry digestive tract its natural reservoir to the consumer’s plate, this pathogen has developed adaptation mechanisms. Among those, biofilm lifestyle has been suggested as a strategy to survive in the food environment and under atmospheric conditions. Recently, the clinical isolate C. jejuni Bf has been shown to survive and grow under aerobic conditions, a property that may help this strain to better survive along the food chain. The aim of this study was to evaluate the adhesion capacity of C. jejuni Bf and its ability to develop a biofilm. C. jejuni Bf can adhere to abiotic surfaces and to human epithelial cells, and can develop biofilm under both microaerobiosis and aerobiosis. These two conditions have no influence on this strain, unlike results obtained with the reference strain C. jejuni 81-176, which harbors only planktonic cells under aerobic conditions. Compared to 81-176, the biofilm of C. jejuni Bf is more homogenous and cell motility at the bottom of biofilm was not modified whatever the atmosphere used. C. jejuni Bf whole genome sequence did not reveal any gene unique to this strain, suggesting that its unusual property does not result from acquisition of new genetic material. Nevertheless some genetic particularities seem to be shared only between Bf and few others strains. Among the main features of C. jejuni Bf genome we noticed (i) a complete type VI secretion system important in pathogenicity and environmental adaptation; (ii) a mutation in the oorD gene involved in oxygen metabolism; and (iii) the presence of an uncommon insertion of a 72 amino acid coding sequence upstream from dnaK, which is involved in stress resistance. Therefore, the atypical behavior of this strain under

  11. Caecal transcriptome analysis of colonized and non-colonized chickens within two genetic lines that differ in caecal colonization by Campylobacter jejuni

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Campylobacter jejuni (C. jejuni) is one of the most common causes of human bacterial enteritis worldwide. The molecular mechanisms of the host responses of chickens to C. jejuni colonization have not been well understood. We have previously shown differences in C. jejuni colonization at 7 days pos...

  12. Method-dependent variability in determination of prevalence of Campylobacter jejuni and Campylobacter coli in Canadian retail poultry.

    PubMed

    Carrillo, Catherine D; Plante, Daniel; Iugovaz, Irène; Kenwell, Robyn; Bélanger, Ghislaine; Boucher, Francine; Poulin, Nathalie; Trottier, Yvon-Louis

    2014-10-01

    Campylobacter is the most frequent cause of bacterial gastroenteritis in Canada, and the illness is commonly associated with poultry consumption. Whereas Canadian retail poultry is often contaminated with campylobacters, studies on the prevalence of this organism are inconsistent due to variability in sampling and microbiological methodology. To determine the current microbiological status of Canadian poultry, and to evaluate two commonly used microbiological methods, 348 raw poultry samples were collected at retail across Canada over a period of 3 years (2007 to 2010) and were analyzed for the presence of thermophilic Campylobacter species. The overall prevalence of Campylobacter spp. was found to be 42.8% by a combination of the two testing methods, with 33.9% of the samples positive for C. jejuni, 3.7% of the samples positive for C. coli, and 5.2% of the samples positive for both. Variability in Campylobacter spp. prevalence was observed in samples obtained from different regions across Canada and from poultry with or without skin, but this was not statistically significant. In co-contaminated samples, C. jejuni was preferentially recovered from Preston agar compared with mCCDA and Campy-Cefex agar, with an increase in recovery of C. coli on all selective media after 48 h of enrichment. A subset of 214 of the poultry rinses were analyzed by both Health Canada's standard method, MFLP-46 (enrichment in Park and Sanders broth), and a second method requiring enrichment in Bolton broth. Significantly more positive samples were obtained with the MFLP-46 method (40.6%) than with the alternate method (35.0%). This improved recovery with MFLP-46 may be due to the omission of cycloheximide from this method. These results demonstrate that determination of prevalence of Campylobacter spp. on poultry products may be significantly impacted by the choice of microbiological methods used. Canadian poultry continues to be a source of exposure to Campylobacter spp.

  13. Campylobacter jejuni is not merely a commensal in commercial broiler chickens and affects bird welfare.

    PubMed

    Humphrey, Suzanne; Chaloner, Gemma; Kemmett, Kirsty; Davidson, Nicola; Williams, Nicola; Kipar, Anja; Humphrey, Tom; Wigley, Paul

    2014-07-01

    Campylobacter jejuni is the leading cause of bacterial food-borne infection; chicken meat is its main source. C. jejuni is considered commensal in chickens based on experimental models unrepresentative of commercial production. Here we show that the paradigm of Campylobacter commensalism in the chicken is flawed. Through experimental infection of four commercial breeds of broiler chickens, we show that breed has a significant effect on C. jejuni infection and the immune response of the animals, although these factors have limited impact on the number of bacteria in chicken ceca. All breeds mounted an innate immune response. In some breeds, this response declined when interleukin-10 was expressed, consistent with regulation of the intestinal inflammatory response, and these birds remained healthy. In another breed, there was a prolonged inflammatory response, evidence of damage to gut mucosa, and diarrhea. We show that bird type has a major impact on infection biology of C. jejuni. In some breeds, infection leads to disease, and the bacterium cannot be considered a harmless commensal. These findings have implications for the welfare of chickens in commercial production where C. jejuni infection is a persistent problem. Importance: Campylobacter jejuni is the most common cause of food-borne bacterial diarrheal disease in the developed world. Chicken is the most common source of infection. C. jejuni infection of chickens had previously not been considered to cause disease, and it was thought that C. jejuni was part of the normal microbiota of birds. In this work, we show that modern rapidly growing chicken breeds used in intensive production systems have a strong inflammatory response to C. jejuni infection that can lead to diarrhea, which, in turn, leads to damage to the feet and legs on the birds due to standing on wet litter. The response and level of disease varied between breeds and is related to regulation of the inflammatory immune response. These findings

  14. Production of a monoclonal antibody specific for the major outer membrane protein of Campylobacter jejuni and characterization of the epitope.

    PubMed

    Qian, Hongliang; Pang, Ervinna; Du, Qingyun; Chang, Jason; Dong, Jin; Toh, Say Ling; Ng, Fook Kheong; Tan, Ai Ling; Kwang, Jimmy

    2008-02-01

    Campylobacter species are important enteric pathogens causing disease in humans and animals. There is a lack of a good immunological test that can be used routinely to separate Campylobacter jejuni from other Campylobacter species. We produced monoclonal antibodies (MAbs) directed against the major outer membrane protein (MOMP) of C. jejuni using recombinant MOMP as the antigen. One MAb, designated MAb5C4 and of the immunoglobulin G1 isotype, was found to be potentially specific for C. jejuni. Dot blots demonstrated that MAb5C4 reacted with all 29 isolates of C. jejuni tested but did not react with 2 C. jejuni isolates, 26 other Campylobacter spp. isolates, and 19 non-Campylobacter isolates. Western blotting showed that MAb5C4 bound to a single protein band approximately 43 kDa in size, corresponding to the expected size of C. jejuni MOMP. The detection limit of MAb5C4 in a dot blot assay was determined to be about 5 x 10(3) bacteria. The epitope on the MOMP was mapped to a region six amino acids in length with the sequence 216GGQFNP221, which is 97% conserved among C. jejuni strains but divergent in other Campylobacter spp.; a GenBank search indicated that 95% of C. jejuni isolates will be able to be detected from non-Campylobacter spp. based on the highly specific and conserved region of the GGQFNP polypeptide. The epitope is predicted to be located in a region that is exposed to the periplasm. MAb5C4 is a potentially specific and sensitive MAb that can be used for the specific detection and identification of C. jejuni.

  15. cj0371: A Novel Virulence-Associated Gene of Campylobacter jejuni

    PubMed Central

    Du, Xueqing; Wang, Nan; Ren, Fangzhe; Tang, Hong; Jiao, Xinan; Huang, Jinlin

    2016-01-01

    Campylobacter jejuni is the major cause of human bacterial diarrhea worldwide. Its pathogenic mechanism remains poorly understood. cj0371 is a novel gene that was uncovered using immunoscreening. There have been no previous reports regarding its function. In this study, we constructed an insertion mutant and complement of this gene in C. jejuni and examined changes in virulence. We observed that the cj0371 mutant showed significantly increased invasion and colonization ability. We also investigated the role of cj0371 in motility, chemotaxis, and growth kinetics to further study its function. We found that the cj0371 mutant displays hypermotility, enhanced chemotaxis, and enhanced growth kinetics. In addition, we localized the Cj0371 protein at the poles of C. jejuni by fluorescence microscopy. We present data that collectively significantly proves our hypothesis that cj0371 is a new virulence-associated gene and through the influence of chemotaxis plays a negative role in C. jejuni pathogenicity. PMID:27471500

  16. cj0371: A Novel Virulence-Associated Gene of Campylobacter jejuni.

    PubMed

    Du, Xueqing; Wang, Nan; Ren, Fangzhe; Tang, Hong; Jiao, Xinan; Huang, Jinlin

    2016-01-01

    Campylobacter jejuni is the major cause of human bacterial diarrhea worldwide. Its pathogenic mechanism remains poorly understood. cj0371 is a novel gene that was uncovered using immunoscreening. There have been no previous reports regarding its function. In this study, we constructed an insertion mutant and complement of this gene in C. jejuni and examined changes in virulence. We observed that the cj0371 mutant showed significantly increased invasion and colonization ability. We also investigated the role of cj0371 in motility, chemotaxis, and growth kinetics to further study its function. We found that the cj0371 mutant displays hypermotility, enhanced chemotaxis, and enhanced growth kinetics. In addition, we localized the Cj0371 protein at the poles of C. jejuni by fluorescence microscopy. We present data that collectively significantly proves our hypothesis that cj0371 is a new virulence-associated gene and through the influence of chemotaxis plays a negative role in C. jejuni pathogenicity.

  17. cj0371: A Novel Virulence-Associated Gene of Campylobacter jejuni.

    PubMed

    Du, Xueqing; Wang, Nan; Ren, Fangzhe; Tang, Hong; Jiao, Xinan; Huang, Jinlin

    2016-01-01

    Campylobacter jejuni is the major cause of human bacterial diarrhea worldwide. Its pathogenic mechanism remains poorly understood. cj0371 is a novel gene that was uncovered using immunoscreening. There have been no previous reports regarding its function. In this study, we constructed an insertion mutant and complement of this gene in C. jejuni and examined changes in virulence. We observed that the cj0371 mutant showed significantly increased invasion and colonization ability. We also investigated the role of cj0371 in motility, chemotaxis, and growth kinetics to further study its function. We found that the cj0371 mutant displays hypermotility, enhanced chemotaxis, and enhanced growth kinetics. In addition, we localized the Cj0371 protein at the poles of C. jejuni by fluorescence microscopy. We present data that collectively significantly proves our hypothesis that cj0371 is a new virulence-associated gene and through the influence of chemotaxis plays a negative role in C. jejuni pathogenicity. PMID:27471500

  18. Investigation on the inhibition of Campylobacter jejuni growth with the applications of some preservatives, medicines, herbs and herb preparations.

    PubMed

    Uradzinski, J; Szteyn, J; Kafel, S

    2002-01-01

    The aim of this study was to determine the influence of chosen preservatives, medicines, herbs and herb preparations on 39 Campylobacter jejuni strains, which were isolated from fresh poultry carcasses. De-Nol and ventrisol were used in the original concentration. Propolis, propolan, artecholin and aukalen were of pharmacies origin. The rest of substances examined were 10% water solutions. Out of 11 substances examined, potassium sorbate, sodium benzoate, De-Nol and ventrisol exerted the strongest inhibitory action on Campylobacter jejuni.

  19. 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.

  20. Seroprevalence in Chickens against Campylobacter jejuni Flagellar Capping Protein (FliD) in Selected Areas of the U.S.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Campylobacter jejuni, a Gram-negative rod, is a zoonotic pathogen associated with human acute bacterial gastroenteritis. Poultry products are regarded as a major source for human infection with this microorganism. We have demonstrated that the flagellar capping protein (FliD) of C. jejuni is highl...

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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...

  2. Campylobacter jejuni Gene Expression in the Chick Cecum: Evidence for Adaptation to a Low-Oxygen Environment

    PubMed Central

    Woodall, C. A.; Jones, M. A.; Barrow, P. A.; Hinds, J.; Marsden, G. L.; Kelly, D. J.; Dorrell, N.; Wren, B. W.; Maskell, D. J.

    2005-01-01

    Transcriptional profiling of Campylobacter jejuni during colonization of the chick cecum identified 59 genes that were differentially expressed in vivo compared with the genes in vitro. The data suggest that C. jejuni regulates electron transport and central metabolic pathways to alter its physiological state during establishment in the chick cecum. PMID:16041056

  3. The Campylobacter jejuni CprRS two-component regulatory system regulates aspects of the cell envelope

    Technology Transfer Automated Retrieval System (TEKTRAN)

    As a leading cause of foodborne bacterial gastroenteritis, Campylobacter jejuni is a significant human pathogen. C. jejuni lives commensally in the gastrointestinal tract of animals, but tolerates variable environments during transit to a susceptible host. A two-component regulatory system, CprRS, w...

  4. Increased incidence of Campylobacter jejuni-associated Guillain-Barré syndromes in the Greater Paris area.

    PubMed

    Sivadon-Tardy, V; Porcher, R; Orlikowski, D; Ronco, E; Gault, E; Roussi, J; Durand, M-C; Sharshar, T; Annane, D; Raphael, J-C; Megraud, F; Gaillard, J-L

    2014-08-01

    The role of Campylobacter jejuni as the triggering agent of Guillain-Barré syndrome (GBS) has not been reassessed since the end of the 1990s in France. We report that the number of C. jejuni-related GBS cases increased continuously between 1996 and 2007 in the Paris region (mean annual increment: 7%, P = 0·007).

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

    PubMed

    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.

  6. 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

  7. Effects of high hydrostatic pressure on Campylobacter jejuni in poultry meat.

    PubMed

    Jackowska-Tracz, A; Tracz, M

    2015-01-01

    Campylobacter jejuni inactivation by high pressure processing (HPP) in poultry meat (chicken breast) was investigated. The pressure was created by high hydrostatic pressure piston-cylinder food processor. Contaminated with C. jejuni (108 CFU g-1) samples of ground poultry meat were hermetically sealed in a polyamide-polyethylene bags and exposed to HPP for 9 different combinations of pressure (200 MPa, 300 MPa and 400 MPa) and time (5 min, 10 min and 15 min). Quantitative bacteriological analysis was carried out in order to determine the number of surviving C. jejuni cells. The obtained results showed that C. jejuni is relatively sensitive to high pressure treatment as compared to other food-borne pathogens. The loss of C. jejuni viability increased in a dose- and time-dependent manner. On the basis of the results, D-values were calculated. For reduction C. jejuni in poultry meat by 6 log units (6D-values), considered as sufficient for consumer protection, the application of 300 MPa for 8.73 min, or 400 MPa for 4.37 min is needed. The linear regression equations, which has been calculated on the basis of this study, allows to determine the degree of C. jejuni reduction in poultry meat for any selected duration of pressurization in a given pressure range.

  8. Salmonella Amager, Campylobacter jejuni, and urease-positive thermophilic Campylobacter found in free-flying peregrine falcons (Falco peregrinus) in Sweden.

    PubMed

    Palmgren, Helena; Broman, Tina; Waldenström, Jonas; Lindberg, Peter; Aspán, Anna; Olsen, Björn

    2004-07-01

    Rare species with small population sizes are vulnerable to perturbations such as disease, inbreeding, or random events. The threat arising from microbial pathogens could be large and other species could act as reservoirs for pathogens. We report finding three enteric bacterial species, Salmonella Amager, Campylobacter jejuni, and urease-positive thermophilic Campylobacter, in nestling free-flying peregrine falcons (Falco peregrinus) in Sweden in 2000. Campylobacter jejuni isolates exhibited marked genetic similarities to an isolate from a human, providing a possible association between a human-associated strain of this bacterium and peregrine falcons.

  9. Mechanisms underlying zoonotic success of Campylobacter jejuni: the CprRS two-component regulatory system influences essential processes, biofilm formation, and pathogenesis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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...

  10. Human antibody response to Campylobacter jejuni flagellin protein and a synthetic N-terminal flagellin peptide.

    PubMed

    Nachamkin, I; Yang, X H

    1989-10-01

    We measured isotype-specific human antibodies directed against Campylobacter jejuni native flagellin and a synthetic peptide derived from the N-terminal amino acid sequence of the protein by using a microdilution enzyme-linked immunosorbent assay (ELISA). Serum samples from patients with gastrointestinal infection caused by C. jejuni (n = 20) and control samples (number from normal subjects = 20; number from patients with diarrhea other than campylobacter = 20) were tested in this assay. Serum specimens from patients with campylobacter infection showed statistically significant higher isotype-specific antiflagellin antibody titers than control samples did. Detection of immunoglobulin G (IgG) antibodies was less specific (70%) than detection of either IgA or IgM antibodies in infected patients (95%). The sensitivity of testing for any of the isotypes ranged from 64 to 100% in acute-phase serum specimens and 85 to 95% in convalescent-phase serum specimens. An ELISA with an N-terminal synthetic peptide derived from the flagellin protein as antigen was not sensitive (60%) for detecting campylobacter infection but was very specific (97.5%). In conclusion, detection of serum IgA or IgM against C. jejuni flagellin may be a useful marker of infection. Although the N-terminal synthetic peptide was antigenic in a few patients with infection and showed good specificity in the ELISA, additional amino acid sequences with better sensitivity for detecting infection need to be identified.

  11. Development of a selective broth medium for the detection of injured Campylobacter jejuni by capacitance monitoring.

    PubMed

    Line, J Eric; Pearson, Kirsten G

    2003-10-01

    The purpose of these studies was to develop a conductimetric method for the rapid detection of Campylobacter jejuni. Numerous basal medium components were analyzed to develop a growth-enhancing broth medium for detection of freeze-injured Campylobacter cells using a conductimetric system. The final medium was composed of a modified Campy-Line agar from which the agar and triphenyltetrazolium chloride were removed and the amino acid, L-arginine was added. Pure isolates of C. jejuni. (frozen and thawed to produce stressed cells) were utilized to test the detection methodology. Monitoring of significant changes in the capacitance signal was found suitable for detection of Campylobacter proliferation. Using stressed pure cultures, Campylobacter growth was repeatedly detected at very low inoculum levels (about one cell per well). There was a direct linear relationship between detection times (DTs) and the initial inoculum level. For example, using a single strain, the mean DT (n = 20) at the 10 CFU/ml inoculum level was 28.6 h, with 100% of the inoculated wells detecting. The mean DTs at the 100, 1,000, and 10,000 CFU/ml inoculum levels were 24.9, 21.4, and 17.0 h, respectively. This study demonstrates that conductimetric methods can be utilized for the rapid detection of C. jejuni.

  12. Effect of propionic acid on Campylobacter jejuni attached to chicken skin during refrigerated storage.

    PubMed

    González-Fandos, Elena; Maya, Naiara; Pérez-Arnedo, Iratxe

    2015-09-01

    The ability of propionic acid to reduce Campylobacter jejuni on chicken legs was evaluated. Chicken legs were inoculated with Campylobacter jejuni. After dipping legs in either water (control), 1% or 2% propionic acid solution (vol/vol), they were stored at 4ºC for 8 days. Changes in C. jejuni, psychrotrophs and Pseudomonas counts were evaluated. Washing in 2% propionic acid significantly reduced C. jejuni counts compared to control legs, with a decrease of about 1.62 log units after treatment. Treatment of chicken legs with 1 or 2% propionic acid significantly reduced numbers of psychrotrophs 1.01 and 1.08 log units and Pseudomonas counts 0.75 and 0.96 log units, respectively, compared to control legs. The reduction in psychrotrophs and Pseudomonas increased throughout storage. The highest reductions obtained for psychrotrophs and Pseudomonas counts in treated legs were reached at the end of storage, day 8, being 3.3 and 2.93 log units, respectively, compared to control legs. Propionic acid treatment was effective in reducing psychrotrophs and Pseudomonas counts on chicken legs throughout storage. It is concluded that propionic acid is effective for reducing C. jejuni populations in chicken. PMID:27036744

  13. Prevalence and characterization of Campylobacter jejuni from chicken meat sold in French retail outlets.

    PubMed

    Guyard-Nicodème, Muriel; Rivoal, Katell; Houard, Emmanuelle; Rose, Valérie; Quesne, Ségolène; Mourand, Gwenaëlle; Rouxel, Sandra; Kempf, Isabelle; Guillier, Laurent; Gauchard, Françoise; Chemaly, Marianne

    2015-06-16

    Campylobacter was detected in 76% of broiler meat products collected in retail outlets during a monitoring plan carried out in France throughout 2009. Campylobacter jejuni was the most prevalent species (64.7% of products being contaminated). The 175 C. jejuni isolates collected were characterized. MLST typing results confirmed substantial genetic diversity as the 175 C. jejuni isolates generated 76 sequence types (STs). The ST-21, ST-45 and ST-464 complexes predominated accounting for 43% of all isolates. A class-specific PCR to screen the sialylated lipooligosaccharide (LOS) locus classes A, B and C showed that 50.3% of the C. jejuni isolates harbored sialylated LOS. The antimicrobial resistance profiles established using a subset of 97 isolates showed that resistance to tetracycline was the most common (53.6%), followed with ciprofloxacin and nalidixic acid (32.9%, and 32.0% respectively). All the tested isolates were susceptible to erythromycin, chloramphenicol and gentamicin. Clear associations were demonstrated between certain clonal complexes and LOS locus classes and between certain clonal complexes and antimicrobial resistance. This work paints a representative picture of C. jejuni isolated from poultry products circulating in France, providing data on STs, LOS locus classes and antibiotic resistance profiles in isolates recovered from products directly available to the consumer.

  14. Effect of propionic acid on Campylobacter jejuni attached to chicken skin during refrigerated storage.

    PubMed

    González-Fandos, Elena; Maya, Naiara; Pérez-Arnedo, Iratxe

    2015-09-01

    The ability of propionic acid to reduce Campylobacter jejuni on chicken legs was evaluated. Chicken legs were inoculated with Campylobacter jejuni. After dipping legs in either water (control), 1% or 2% propionic acid solution (vol/vol), they were stored at 4ºC for 8 days. Changes in C. jejuni, psychrotrophs and Pseudomonas counts were evaluated. Washing in 2% propionic acid significantly reduced C. jejuni counts compared to control legs, with a decrease of about 1.62 log units after treatment. Treatment of chicken legs with 1 or 2% propionic acid significantly reduced numbers of psychrotrophs 1.01 and 1.08 log units and Pseudomonas counts 0.75 and 0.96 log units, respectively, compared to control legs. The reduction in psychrotrophs and Pseudomonas increased throughout storage. The highest reductions obtained for psychrotrophs and Pseudomonas counts in treated legs were reached at the end of storage, day 8, being 3.3 and 2.93 log units, respectively, compared to control legs. Propionic acid treatment was effective in reducing psychrotrophs and Pseudomonas counts on chicken legs throughout storage. It is concluded that propionic acid is effective for reducing C. jejuni populations in chicken.

  15. 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

  16. Potential competitive exclusion bacteria from poultry inhibitory to Campylobacter jejuni and Salmonella.

    PubMed

    Zhang, Guodong; Ma, Li; Doyle, Michael P

    2007-04-01

    The objective of this study was to isolate from chickens potential competitive exclusion bacteria (CE) that are inhibitory to Campylobacter jejuni or Salmonella, or to both, for subsequent development of a defined CE product for use in poultry. Adult chickens from family farms, commercial farms, and broiler chicken research centers were sampled to identify and select C. jejuni-free donor chickens. A challenge treatment, which included administering perorally 106 CFU C. jejuni per chicken and determining undetectable cecal shedding of campylobacters at 4 weeks, was important for identifying the best CE donor chickens. Screening of bacterial colonies obtained from nine donor chickens by using selective and nonselective media yielded 636 isolates inhibitory to six C. jejuni strains in vitro, with 194 isolates being strongly inhibitory. Of the 194 isolates, 145 were from ceca, and 117 were facultative anaerobic bacteria. One hundred forty-three isolates were inhibitory to six strains of Salmonella (including five different serotypes) in vitro. Of these, 41 were strongly inhibitory to all C. jejuni and Salmonella strains evaluated, and most were Lactobacillus salivarius. A direct overlay method, which involved directly applying soft agar on plates with discrete colonies from mucus scrapings of gastrointestinal tracts, was more effective in isolating CE than was the frequently practiced isolation method of picking and transferring discrete colonies and then overlaying them with soft agar. The best approach for obtaining bacteria highly inhibitory to Salmonella and C. jejuni from chickens was to isolate bacteria from ceca under anaerobic conditions. Free-range chickens from family farms were better donors of potential CE strongly inhibitory to both Salmonella and Campylobacter than were chickens from commercial farms and broiler chicken research centers.

  17. Association of Campylobacter Jejuni ssp. Jejuni Chemotaxis Receptor Genes with Multilocus Sequence Types and Source of Isolation

    PubMed Central

    Mund, Norah Lynn-Anne; Masanta, Wycliffe Omurwa; Goldschmidt, Anne-Marie; Lugert, Raimond; Groß, Uwe; Zautner, Andreas E.

    2016-01-01

    Campylobacter jejuni’s flagellar locomotion is controlled by eleven chemoreceptors. Assessment of the distribution of the relevant chemoreceptor genes in the C. jejuni genomes deposited in the National Center for Biotechnology Information (NCBI) database led to the identification of two previously unknown tlp genes and a tlp5 pseudogene. These two chemoreceptor genes share the same locus in the C. jejuni genome with tlp4 and tlp11, but the gene region encoding the periplasmic ligand binding domain differs significantly from other chemoreceptor genes. Hence, they were named tlp12 and tlp13. Consequently, it was of interest to study their distribution in C. jejuni subpopulations of different clonality, and their cooccurrence with the eleven previously reported chemoreceptor genes. Therefore, the presence of all tlp genes was detected by polymerase chain reaction (PCR) in 292 multilocus sequence typing (MLST)-typed C. jejuni isolates from different hosts. The findings show interesting trends: Tlp4, tlp11, tlp12, and tlp13 appeared to be mutually exclusive and cooccur in a minor subset of isolates. Tlp4 was found to be present in only 33.56% of all tested isolates and was significantly less often detected in turkey isolates. Tlp11 was tested positive in only 17.8% of the isolates, while tlp12 was detected in 29.5% of all isolates, and tlp13 was found to be present in 38.7%. PMID:27766165

  18. Genetic Diversity of Campylobacter jejuni and Campylobacter coli Isolates from Conventional Broiler Flocks and the Impacts of Sampling Strategy and Laboratory Method

    PubMed Central

    Colles, F. M.; Rodgers, J. D.; McCarthy, N. D.; Davies, R. H.; Maiden, M. C. J.; Clifton-Hadley, F. A.

    2016-01-01

    The genetic diversity of Campylobacter jejuni and Campylobacter coli isolates from commercial broiler farms was examined by multilocus sequence typing (MLST), with an assessment of the impact of the sample type and laboratory method on the genotypes of Campylobacter isolated. A total of 645 C. jejuni and 106 C. coli isolates were obtained from 32 flocks and 17 farms, with 47 sequence types (STs) identified. The Campylobacter jejuni isolates obtained by different sampling approaches and laboratory methods were very similar, with the same STs identified at similar frequencies, and had no major effect on the genetic profile of Campylobacter population in broiler flocks at the farm level. For C. coli, the results were more equivocal. While some STs were widely distributed within and among farms and flocks, analysis of molecular variance (AMOVA) revealed a high degree of genetic diversity among farms for C. jejuni, where farm effects accounted for 70.5% of variance, and among flocks from the same farm (9.9% of variance for C. jejuni and 64.1% for C. coli). These results show the complexity of the population structure of Campylobacter in broiler production and that commercial broiler farms provide an ecological niche for a wide diversity of genotypes. The genetic diversity of C. jejuni isolates among broiler farms should be taken into account when designing studies to understand Campylobacter populations in broiler production and the impact of interventions. We provide evidence that supports synthesis of studies on C. jejuni populations even when laboratory and sampling methods are not identical. PMID:26873321

  19. Analysis of Campylobacter jejuni whole genome DNA microarrays: significance of prophage and hypervariable regions for discriminating isolates

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Introduction: Campylobacter jejuni is a major cause of gastroenteritis in humans and is carried in many common food animals. In order to reduce human infections a better understanding of Campylobacter epidemiology is needed. Identifying genes that enable discriminating between isolates is an importa...

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

    PubMed Central

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

    2016-01-01

    Introduction 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. Materials and methods 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. Results and discussion 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

  1. The Campylobacter jejuni Ferric Uptake Regulator Promotes Acid Survival and Cross-Protection against Oxidative Stress

    PubMed Central

    Askoura, Momen; Sarvan, Sabina; Couture, Jean-François

    2016-01-01

    Campylobacter jejuni is a prevalent cause of bacterial gastroenteritis in humans worldwide. The mechanisms by which C. jejuni survives stomach acidity remain undefined. In the present study, we demonstrated that the C. jejuni ferric uptake regulator (Fur) plays an important role in C. jejuni acid survival and acid-induced cross-protection against oxidative stress. A C. jejuni Δfur mutant was more sensitive to acid than the wild-type strain. Profiling of the acid stimulon of the C. jejuni Δfur mutant allowed us to uncover Fur-regulated genes under acidic conditions. In particular, Fur was found to upregulate genes involved in flagellar and cell envelope biogenesis upon acid stress, and mutants with deletions of these genes were found to be defective in surviving acid stress. Interestingly, prior acid exposure of C. jejuni cross-protected against oxidative stress in a catalase (KatA)- and Fur-dependent manner. Western blotting and reverse transcription-quantitative PCR revealed increased expression of KatA upon acid stress. Electrophoretic mobility shift assays (EMSAs) demonstrated that the binding affinity between Fur and the katA promoter is reduced in vitro under conditions of low pH, rationalizing the higher levels of expression of katA under acidic conditions. Strikingly, the Δfur mutant exhibited reduced virulence in both human epithelial cells and the Galleria mellonella infection model. Altogether, this is the first study showing that, in addition to its role in iron metabolism, Fur is an important regulator of C. jejuni acid responses and this function cross-protects against oxidative stress. Moreover, our results clearly demonstrate Fur's important role in C. jejuni pathogenesis. PMID:26883589

  2. Campylobacter jejuni cell lysates differently target mitochondria and lysosomes on HeLa cells.

    PubMed

    Canonico, B; Campana, R; Luchetti, F; Arcangeletti, M; Betti, M; Cesarini, E; Ciacci, C; Vittoria, E; Galli, L; Papa, S; Baffone, W

    2014-08-01

    Campylobacter jejuni is the most common cause of bacterial gastroenteritis in humans. The synthesis of cytolethal distending toxin appears essential in the infection process. In this work we evaluated the sequence of lethal events in HeLa cells exposed to cell lysates of two distinct strains, C. jejuni ATCC 33291 and C. jejuni ISS3. C. jejuni cell lysates (CCLys) were added to HeLa cell monolayers which were analysed to detect DNA content, death features, bcl-2 and p53 status, mitochondria/lysosomes network and finally, CD54 and CD59 alterations, compared to cell lysates of C. jejuni 11168H cdtA mutant. We found mitochondria and lysosomes differently targeted by these bacterial lysates. Death, consistent with apoptosis for C. jejuni ATCC 33291 lysate, occurred in a slow way (>48 h); concomitantly HeLa cells increase their endolysosomal compartment, as a consequence of toxin internalization besides a simultaneous and partial lysosomal destabilization. C. jejuni CCLys induces death in HeLa cells mainly via a caspase-dependent mechanism although a p53 lysosomal pathway (also caspase-independent) seems to appear in addition. In C. jejuni ISS3-treated cells, the p53-mediated oxidative degradation of mitochondrial components seems to be lost, inducing the deepest lysosomal alterations. Furthermore, CD59 considerably decreases, suggesting both a degradation or internalisation pathway. CCLys-treated HeLa cells increase CD54 expression on their surface, because of the action of lysate as its double feature of toxin and bacterial peptide. In conclusion, we revealed that C. jejuni CCLys-treated HeLa cells displayed different features, depending on the particular strain.

  3. 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

    2012-07-01

    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

  4. Identification of Campylobacter jejuni ATCC 43431-Specific Genes by Whole Microbial Genome Comparisons

    PubMed Central

    Poly, Frédéric; Threadgill, Deborah; Stintzi, Alain

    2004-01-01

    This study describes a novel approach to identify unique genomic DNA sequences from the unsequenced strain C. jejuni ATCC 43431 by comparison with the sequenced strain C. jejuni NCTC 11168. A shotgun DNA microarray was constructed by arraying 9,600 individual DNA fragments from a C. jejuni ATCC 43431 genomic library onto a glass slide. DNA fragments unique to C. jejuni ATCC 43431 were identified by competitive hybridization to the array with genomic DNA of C. jejuni NCTC 11168. The plasmids containing unique DNA fragments were sequenced, allowing the identification of up to 130 complete and incomplete genes. Potential biological roles were assigned to 66% of the unique open reading frames. The mean G+C content of these unique genes (26%) differs significantly from the G+C content of the entire C. jejuni genome (30.6%). This suggests that they may have been acquired through horizontal gene transfer from an organism with a G+C content lower than that of C. jejuni. Because the two C. jejuni strains differ by Penner serotype, a large proportion of the unique ATCC 43431 genes encode proteins involved in lipooligosaccharide and capsular biosynthesis, as expected. Several unique open reading frames encode enzymes which may contribute to genetic variability, i.e., restriction-modification systems and integrases. Interestingly, many of the unique C. jejuni ATCC 43431 genes show identity with a possible pathogenicity island from Helicobacter hepaticus and components of a potential type IV secretion system. In conclusion, this study provides a valuable resource to further investigate Campylobacter diversity and pathogenesis. PMID:15231810

  5. Prospective study of Campylobacter jejuni infection in Chilean infants evaluated by culture and serology.

    PubMed

    Figueroa, G; Galeno, H; Troncoso, M; Toledo, S; Soto, V

    1989-05-01

    A prospective study of Campylobacter jejuni infection was performed during a 6-month period in a cohort of 198 Chilean infants. Surveillance was based on biweekly home visits by a team of trained nurses. C. jejuni colonization was studied by culturing stool samples from diarrheal episodes and paired asymptomatic controls and by performing an enzyme-linked immunosorbent assay developed to measure C. jejuni outer membrane protein polyvalent antibodies in a representative group of infants. C. jejuni was isolated from 30 of 299 diarrheal episodes (10%) and from 17 of 304 samples from asymptomatic infants (6%). Significantly higher (P less than 0.05) C. jejuni illness/infection ratios were found in infants less than 9 months old. Polyvalent antibodies to C. jejuni were found in 27 of 89 serum samples (30%). The lowest prevalence (4%) was observed among infants less than 9 months old, and the highest was found in patients older than 15 months (63%; P less than 0.01). Data obtained by analysis of paired serum samples revealed a significant increase in the number of seropositive individuals, from 8% on admission to 50% at the end of the protocol (P less than 0.001). The change in the immune status of these infants was associated with symptomatic infection in 9 of 18 cases. The antibody rise found in the remaining nine infants suggested the presence of asymptomatic C. jejuni infections. This inference was documented by the isolation of C. jejuni from stools of two of these infants when the infants were randomly studied while asymptomatic. This study points out the impact of C. jejuni-associated diarrheal episodes and the high frequency with which asymptomatic infections elicit specific antibodies in infants living in areas such as Chile.

  6. Evaluation of passive immunotherapeutic efficacy of hyperimmunized egg yolk powder against intestinal colonization of Campylobacter jejuni in chickens.

    PubMed

    Paul, Narayan C; Al-Adwani, Salma; Crespo, Rocio; Shah, Devendra H

    2014-11-01

    Campylobacter jejuni is a leading cause of foodborne bacterial gastroenteritis in human. Chickens are the reservoir host of C. jejuni, and contaminated chicken meat is an important source of human infection. Therefore, control of C. jejuni in chickens can have direct effect on human health. In this study we tested the passive immunotherapeutic efficacy of the chicken egg-yolk-derived antibodies, in the form of hyperimmunized egg yolk powder (HEYP), against 7 colonization-associated proteins of C. jejuni, namely, CadF (Campylobacter adhesion to fibronectin), FlaA (flagellar proteins), MOMP (major outer membrane protein), FlpA (fibronectin binding protein A), CmeC (Campylobacter multidrug efflux C), Peb1A (Campylobacter putative adhesion), and JlpA (Jejuni lipoprotein A). Three chicken experiments were performed. In each experiment, chickens were treated orally via feed supplemented with 10% (wt/wt) egg yolk powder. In experiment 1, chicken groups were experimentally infected with C. jejuni (10(8) cfu) followed by treatment with 5 HEYP (CadF, FlaA, MOMP, FlpA, CmeC) for 4 d either individually or as a cocktail containing equal parts of each HEYP. In experiment 2, chickens were treated for 21 d with cocktail containing equal parts of 7 HEYP before and after experimental infection with C. jejuni (10(8) cfu). In experiment 3, chickens were treated with feed containing a cocktail of 7 HEYP before and after (prophylaxis), and after (treatment) experimental infection with C. jejuni (10(5) cfu). Intestinal colonization of C. jejuni was monitored by culturing cecal samples from chickens euthanized at the end of each experiment. The results showed that there were no differences in the cecal colonization of C. jejuni between HEYP treated and nontreated control chickens, suggesting that use of HEYP at the dose and the regimens used in the current study is not efficacious in reducing C. jejuni colonization in chickens.

  7. Campylobacter jejuni gene cj0511 encodes a serine peptidase essential for colonisation

    PubMed Central

    Karlyshev, A.V.; Thacker, G.; Jones, M.A.; Clements, M.O.; Wren, B.W.

    2014-01-01

    According to MEROPS peptidase database, Campylobacter species encode 64 predicted peptidases. However, proteolytic properties of only a few of these proteins have been confirmed experimentally. In this study we identified and characterised a Campylobacter jejuni gene cj0511 encoding a novel peptidase. The proteolytic activity associated with this enzyme was demonstrated in cell lysates. Moreover, enzymatic studies conducted with a purified protein confirmed a prediction of it being a serine peptidase. Furthermore, cj0511 mutant was found to be severely attenuated in chicken colonisation model, suggesting a role of the Cj0511 protein in infection. PMID:24918062

  8. Incidence and ecology of Campylobacter jejuni and coli in animals

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Since its initial emergence in the 1970’s, Campylobacter have been estimated to be one of the most common causative agents of foodborne illnesses, along with nontyphoidal Salmonella species. Campylobacter species naturally colonize the gastrointestinal tracts of domestic and feral animals and are a...

  9. Supplementation of Bolton broth with triclosan improves detection of Campylobacter jejuni and Campylobacter coli in chicken carcass rinse.

    PubMed

    Chon, Jung-Whan; Kim, Young-Ji; Kim, Hong-Seok; Kim, Dong-Hyeon; Kim, Hyunsook; Song, Kwang-Young; Seo, Kun-Ho

    2014-07-01

    We compared Bolton enrichment broth supplemented with antimicrobial triclosan (T-Bolton broth) and normal Bolton broth for the isolation of Campylobacter jejuni (C. jejuni) and Campylobacter coli (C. coli) from chicken carcass rinse. Whole chickens were rinsed with buffered peptone water prior to enrichment in normal Bolton broth or T-Bolton broth, followed by inoculation onto modified charcoal-cefoperazone-deoxycholate agar (mCCDA). Suspect colonies were confirmed by PCR. We observed a significantly higher number of C. jejuni or C. coli-positive samples in the T-Bolton broth (71.3%) than in the normal Bolton broth (27.5%) (p<0.05). Furthermore, the number of contaminated mCCDA plates was lower after enrichment in T-Bolton broth (3.8%) than in the normal Bolton broth (75%) (p<0.05), indicating that T-Bolton broth has higher selectivity. Finally, we identified extended-spectrum β-lactamase-producing Escherichia coli as the predominant competing flora in normal Bolton broth. In conclusion, the use of T-Bolton broth results in significant elimination of competing bacteria.

  10. Identification of the main quinolone resistance determinant in Campylobacter jejuni and Campylobacter coli by MAMA-DEG PCR.

    PubMed

    Hormeño, Lorena; Palomo, Gonzalo; Ugarte-Ruiz, María; Porrero, M Concepción; Borge, Carmen; Vadillo, Santiago; Píriz, Segundo; Domínguez, Lucas; Campos, Maria J; Quesada, Alberto

    2016-03-01

    Among zoonotic diseases, campylobacteriosis stands out as the major bacterial infection producing human gastroenteritis. Antimicrobial therapy, only recommended in critical cases, is challenged by resistance mechanisms that should be unambiguously detected for achievement of effective treatments. Quinolone (ciprofloxacin) resistance of Campylobacter jejuni and Campylobacter coli, the 2 main Campylobacter detected in humans, is conferred by the mutation gyrA C-257-T, which can be genotyped by several methods that require a previous identification of the pathogen species to circumvent the sequence polymorphism of the gene. A multiplex PCR, based on degenerated oligonucleotides, has been designed for unambiguous identification of the quinolone resistance determinant in Campylobacter spp. isolates. The method was verified with 249 Campylobacter strains isolated from humans (141 isolates) and from the 3 most important animal sources for this zoonosis: poultry (34 isolates), swine (38 isolates), and cattle (36 isolates). High resistance to ciprofloxacin, MIC above 4μg/mL, linked to the mutated genotype predicted by MAMA-DEG PCR (mismatch amplification mutation assay PCR with degenerated primers) was found frequently among isolates from the different hosts.

  11. Role of flgA for Flagellar Biosynthesis and Biofilm Formation of Campylobacter jejuni NCTC11168.

    PubMed

    Kim, Joo-Sung; Park, Changwon; Kim, Yun-Ji

    2015-11-01

    The complex roles of flagella in the pathogenesis of Campylobacter jejuni, a major cause of worldwide foodborne diarrheal disease, are important. Compared with the wild-type, an insertional mutation of the flgA gene (cj0769c) demonstrated significant decrease in the biofilm formation of C. jejuni NCTC11168 on major food contact surfaces, such as polystyrene, stainless steel, and borosilicate glass. The flgA mutant was completely devoid of flagella and non-motile whereas the wild-type displayed the full-length flagella and motility. In addition, the biofilm formation of the wild-type was inversely dependent on the viscosity of the media. These results support that flagellar-mediated motility plays a significant role in the biofilm formation of C. jejuni NCTC11168. Moreover, our adhesion assay suggests that it plays an important role during biofilm maturation after initial attachment. Furthermore, C. jejuni NCTC11168 wild-type formed biofilm with a net-like structure of extracellular fiber-like material, but such a structure was significantly reduced in the biofilm of the flgA mutant. It supports that the extracellular fiber-like material may play a significant role in the biofilm formation of C. jejuni. This study demonstrated that flgA is essential for flagellar biosynthesis and motility, and plays a significant role in the biofilm formation of C. jejuni NCTC11168.

  12. Chicken Immune Response after In Ovo Immunization with Chimeric TLR5 Activating Flagellin of Campylobacter jejuni

    PubMed Central

    Radomska, Katarzyna A.; Vaezirad, Mahdi M.; Verstappen, Koen M.; Wösten, Marc M. S. M.; Wagenaar, Jaap A.; van Putten, Jos P. M.

    2016-01-01

    Campylobacter jejuni is the main cause of bacterial food-borne diseases in developed countries. Chickens are the most important source of human infection. Vaccination of poultry is an attractive strategy to reduce the number of C. jejuni in the intestinal tract of chickens. We investigated the immunogenicity and protective efficacy of a recombinant C. jejuni flagellin-based subunit vaccine with intrinsic adjuvant activity. Toll-like receptor activation assays demonstrated the purity and TLR5 stimulating (adjuvant) activity of the vaccine. The antigen (20–40 μg) was administered in ovo to 18 day-old chicken embryos. Serum samples and intestinal content were assessed for antigen-specific systemic and mucosal humoral immune responses. In ovo vaccination resulted in the successful generation of IgY and IgM serum antibodies against the flagellin-based subunit vaccine as determined by ELISA and Western blotting. Vaccination did not induce significant amounts of flagellin-specific secretory IgA in the chicken intestine. Challenge of chickens with C. jejuni yielded similar intestinal colonization levels for vaccinated and control animals. Our results indicate that in ovo delivery of recombinant C. jejuni flagellin subunit vaccine is a feasible approach to yield a systemic humoral immune response in chickens but that a mucosal immune response may be needed to reduce C. jejuni colonization. PMID:27760175

  13. Campylobacter jejuni Motility Is Required for Infection of the Flagellotropic Bacteriophage F341

    PubMed Central

    Baldvinsson, Signe Berg; Sørensen, Martine C. Holst; Vegge, Christina S.; Clokie, Martha R. J.

    2014-01-01

    Previous studies have identified a specific modification of the capsular polysaccharide as receptor for phages that infect Campylobacter jejuni. Using acapsular kpsM mutants of C. jejuni strains NCTC11168 and NCTC12658, we found that bacteriophage F341 infects C. jejuni independently of the capsule. In contrast, phage F341 does not infect C. jejuni NCTC11168 mutants that either lack the flagellar filaments (ΔflaAB) or that have paralyzed, i.e., nonrotating, flagella (ΔmotA and ΔflgP). Complementing flgP confirmed that phage F341 requires rotating flagella for successful infection. Furthermore, adsorption assays demonstrated that phage F341 does not adsorb to these nonmotile C. jejuni NCTC11168 mutants. Taken together, we propose that phage F341 uses the flagellum as a receptor. Phage-host interactions were investigated using fluorescence confocal and transmission electron microscopy. These data demonstrate that F341 binds to the flagellum by perpendicular attachment with visible phage tail fibers interacting directly with the flagellum. Our data are consistent with the movement of the C. jejuni flagellum being required for F341 to travel along the filament to reach the basal body of the bacterium. The initial binding to the flagellum may cause a conformational change of the phage tail that enables DNA injection after binding to a secondary receptor. PMID:25261508

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

    PubMed Central

    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

    2016-01-01

    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. PMID:27762269

  15. Antimicrobial susceptibilities of Campylobacter jejuni and Campylobacter coli strains isolated from two early stages of poultry production.

    PubMed

    Pérez-Boto, David; García-Peña, Francisco Javier; Abad-Moreno, Juan Carlos; Echeita, M Aurora

    2013-08-01

    Our aim was to monitor the resistance of Campylobacter isolates from two initial stages of broiler production in 5 grandparent breeder broiler farms (GPBFs) and 12 parent breeder broiler farms (PBFs) in which no antimicrobials were used during the study. Susceptibility tests were carried out for 805 strains (697 Campylobacter jejuni and 108 Campylobacter coli) against nalidixic acid, ciprofloxacin, erythromycin, amoxicillin, amoxicillin plus clavulanic acid, tetracycline, gentamicin, and chloramphenicol using the disk-diffusion method. Quinolone resistance was the most abundant overall (74.9%) and at each stage of production. The second largest resistance was for tetracycline with 48.2%. The resistance against amoxicillin plus clavulanic acid, gentamicin, and chloramphenicol was not found. The percentages of resistance and multidrug-resistant (MDR) isolates were always higher in the PBFs than in the GPBFs. However, pan-susceptible populations (total 10.3%) were isolated in our survey. C. coli isolates were more resistant to tetracycline and erythromycin (96.3% and 23.1%, respectively) than for C. jejuni (40.7% and 0%, respectively) and were more MDR (33.3% vs. 11.9%). In conclusion, as other authors have shown, even in the absence of antibiotic pressure, relatively high rates of quinolone resistance are found in Campylobacter. However, a decrease in quinolone resistance has been observed compared to other studies in Spain [i.e., 99%; Saenz et al. Antimicrob. Agents Chemother. 2000;44(2):267-271]. MDR, fluoroquinolone-, macrolide-, and tetracycline-resistant Campylobacter populations are issues of concern in public health.

  16. Identification and characterization of an immunogenic outer membrane protein of Campylobacter jejuni.

    PubMed Central

    Burnens, A; Stucki, U; Nicolet, J; Frey, J

    1995-01-01

    We cloned and expressed in Escherichia coli a gene encoding an 18-kDa outer membrane protein (Omp18) from Campylobacter jejuni ATCC 29428. The nucleotide sequence of the gene encoding Omp18 was determined, and an open reading frame of 165 amino acids was revealed. The amino acid sequence had the typical features of a leader sequence and a signal peptidase II cleavage site at the N-terminal part of Omp18. Moreover, the sequence had a high degree of similarity to the peptidoglycan-associated outer membrane lipoprotein P6 of Haemophilus influenzae and the peptidoglycan-associated lipoprotein PAL of E. coli. Southern blot analysis in which the cloned gene was used as a probe revealed genes similar to that encoding Omp18 in all species of the thermophilic group of campylobacters as well as Campylobacter sputorum. All campylobacters tested expressed a protein with a molecular mass identical to that of Omp18. The protein reacted immunologically with polyclonal antibodies directed against Omp18 from C. jejuni. PCR amplification of the gene encoding Omp18 with specific primers and subsequent restriction enzyme analysis of the amplified DNA fragments showed that the gene for Omp18 is highly conserved in C. jejuni strains isolated from humans, dogs, cats, calves, and chickens but is different in other Campylobacter species. In order to obtain pure recombinant Omp18 protein for serological assays, the cloned gene for Omp18 was genetically modified by replacing the signal sequence with a DNA segment encoding six adjacent histidine residues. Expression of this construct in E. coli allowed purification of the modified protein (Omp18-6xHis) by metal chelation chromatography. Sera from patients with past C. jejuni infection reacted positively with Omp18-6xHis, while sera from healthy blood donors showed no reaction with this antigen. Omp18, which is an outer membrane protein belonging to the family of PALs is well conserved in C. jejuni and is highly immunogenic. It is therefore a

  17. Cloning, sequencing, and characterization of the lipopolysaccharide biosynthetic enzyme heptosyltransferase I gene (waaC) from Campylobacter jejuni and Campylobacter coli.

    PubMed

    Klena, J D; Gray, S A; Konkel, M E

    1998-11-19

    Campylobacter jejuni and Campylobacter coli are common causes of gastrointestinal disease and a proportion of C. jejuni infections have been shown to be associated with the Guillain-Barré syndrome. The waaC gene from Campylobacter coli, involved in lipopolysaccharide core biosynthesis, was cloned by complementation of a heptose-deficient strain of Salmonella typhimurium, as judged by novobiocin sensitivity, lipopolysaccharide (LPS)-specific phage sensitivity, and polyacrylamide-resolved lipopolysaccharide profiles. The C. jejuni waaC gene was subsequently cloned using the waaC gene isolated from C. coli as a probe. The C. jejuni and C. coli waaC genes are capable of encoding proteins of 342 amino acids with calculated molecular masses of 39381Da and 39317Da, respectively. Sequence and in-vitro analyses suggested that the C. coli waaC gene may be transcribed from its own promoter. Translation of the C. coli waaC gene in a cell-free system yielded a protein with a Mr of 39000. The waaC gene was detected in every C. jejuni and C. coli isolate tested as judged by dot-blot hybridization analysis. Southern hybridization analysis indicated that both Campylobacter species contain a single copy of the waaC gene. Unlike Escherichia coli and S. typhimurium isolates, the waaC gene in C. jejuni and C. coli isolates does not appear to be linked to the waaF (rfaF) gene.

  18. The abundant free-living amoeba, Acanthamoeba polyphaga, increases the survival of Campylobacter jejuni in milk and orange juice

    PubMed Central

    Olofsson, Jenny; Berglund, Petra Griekspoor; Olsen, Björn; Ellström, Patrik; Axelsson-Olsson, Diana

    2015-01-01

    Background Campylobacter jejuni is a common cause of human bacterial diarrhea in most parts of the world. Most C. jejuni infections are acquired from contaminated poultry, milk, and water. Due to health care costs and human suffering, it is important to identify all possible sources of infection. Unpasteurized milk has been associated with several outbreaks of C. jejuni infection. Campylobacter has been identified on fresh fruit, and other gastrointestinal pathogens such as Salmonella, E. coli O157:H7 and Cryptosporidium have been involved in fruit juice outbreaks. C. jejuni is sensitive to the acidic environment of fruit juice, but co-cultures with the amoeba, Acanthamoeba polyphaga, have previously been shown to protect C. jejuni at low pH. Methods To study the influence of A. polyphaga on the survival of C. jejuni in milk and juice, the bacteria were incubated in the two products at room temperature and at 4°C with the following treatments: A) C. jejuni preincubated with A. polyphaga before the addition of product, B) C. jejuni mixed with A. polyphaga after the addition of product, and C) C. jejuni in product without A. polyphaga. Bacterial survival was assessed by colony counts on blood agar plates. Results Co-culture with A. polyphaga prolonged the C. jejuni survival both in milk and juice. The effect of co-culture was most pronounced in juice stored at room temperature. On the other hand, A. polyphaga did not have any effect on C. jejuni survival during pasteurization of milk or orange juice, indicating that this is a good method for eliminating C. jejuni in these products. Conclusion Amoebae-associated C. jejuni in milk and juice might cause C. jejuni infections. PMID:26387556

  19. Campylobacter jejuni Colonization Is Associated with a Dysbiosis in the Cecal Microbiota of Mice in the Absence of Prominent Inflammation

    PubMed Central

    Lone, Abdul G.; Selinger, L. Brent; Uwiera, Richard R. E.; Xu, Yong; Inglis, G. Douglas

    2013-01-01

    Background Campylobacter jejuni causes enterocolitis in humans, but does not incite disease in asymptomatic carrier animals. To survive in the intestine, C. jejuni must successfully compete with the microbiota and overcome the host immune defense. Campylobacter jejuni colonization success varies considerably amongst individual mice, and we examined the degree to which the intestinal microbiota was affected in mice (i.e. a model carrier animal) colonized by C. jejuni at high relative to low densities. Methods Mice were inoculated with C. jejuni or buffer, and pathogen shedding and intestinal colonization were measured. Histopathologic scoring and quantification of mRNA expression for α-defensins, toll-like receptors, and cytokine genes were conducted. Mucosa-associated bacterial communities were characterized by two approaches: multiplexed barcoded pyrosequencing and terminal restriction fragment length polymorphism analysis. Results Two C. jejuni treatments were established based on the degree of cecal and colonic colonization; C. jejuni Group A animals were colonized at high cell densities, and C. jejuni Group B animals were colonized at lower cell densities. Histological examination of cecal and colonic tissues indicated that C. jejuni did not incite visible pathologic changes. Although there was no significant difference among treatments in expression of mRNA for α-defensins, toll-like receptors, or cytokine genes, a trend for increased expression of toll-like receptors and cytokine genes was observed for C. jejuni Group A. The results of the two methods to characterize bacterial communities indicated that the composition of the cecal microbiota of C. jejuni Group A mice differed significantly from C. jejuni Group B and Control mice. This difference was due to a reduction in load, diversity and richness of bacteria associated with the cecal mucosa of C. jejuni Group A mice. Conclusions High density colonization by C. jejuni is associated with a dysbiosis in

  20. Superoxide dismutase SodB is a protective antigen against Campylobacter jejuni colonisation in chickens.

    PubMed

    Chintoan-Uta, Cosmin; Cassady-Cain, Robin L; Al-Haideri, Halah; Watson, Eleanor; Kelly, David J; Smith, David G E; Sparks, Nick H C; Kaiser, Pete; Stevens, Mark P

    2015-11-17

    Campylobacter is the leading cause of foodborne diarrhoeal illness in the developed world and consumption or handling of contaminated poultry meat is the principal source of infection. Strategies to control Campylobacter in broilers prior to slaughter are urgently required and are predicted to limit the incidence of human campylobacteriosis. Towards this aim, a purified recombinant subunit vaccine based on the superoxide dismutase (SodB) protein of C. jejuni M1 was developed and tested in White Leghorn birds. Birds were vaccinated on the day of hatch and 14 days later with SodB fused to glutathione S-transferase (GST) or purified GST alone. Birds were challenged with C. jejuni M1 at 28 days of age and caecal Campylobacter counts determined at weekly intervals. Across three independent trials, the vaccine induced a statistically significant 1 log10 reduction in caecal Campylobacter numbers in vaccinated birds compared to age-matched GST-vaccinated controls. Significant induction of antigen-specific serum IgY was detected in all vaccinated birds, however the magnitude and timing of SodB-specific IgY did not correlate with lower numbers of C. jejuni. Antibodies from SodB-vaccinated chickens detected the protein in the periplasm and not membrane fractions or on the bacterial surface, suggesting that the protection observed may not be strictly antibody-mediated. SodB may be useful as a constituent of vaccines for control of C. jejuni infection in broiler birds, however modest protection was observed late relative to the life of broiler birds and further studies are required to potentiate the magnitude and timing of protection.

  1. Superoxide dismutase SodB is a protective antigen against Campylobacter jejuni colonisation in chickens.

    PubMed

    Chintoan-Uta, Cosmin; Cassady-Cain, Robin L; Al-Haideri, Halah; Watson, Eleanor; Kelly, David J; Smith, David G E; Sparks, Nick H C; Kaiser, Pete; Stevens, Mark P

    2015-11-17

    Campylobacter is the leading cause of foodborne diarrhoeal illness in the developed world and consumption or handling of contaminated poultry meat is the principal source of infection. Strategies to control Campylobacter in broilers prior to slaughter are urgently required and are predicted to limit the incidence of human campylobacteriosis. Towards this aim, a purified recombinant subunit vaccine based on the superoxide dismutase (SodB) protein of C. jejuni M1 was developed and tested in White Leghorn birds. Birds were vaccinated on the day of hatch and 14 days later with SodB fused to glutathione S-transferase (GST) or purified GST alone. Birds were challenged with C. jejuni M1 at 28 days of age and caecal Campylobacter counts determined at weekly intervals. Across three independent trials, the vaccine induced a statistically significant 1 log10 reduction in caecal Campylobacter numbers in vaccinated birds compared to age-matched GST-vaccinated controls. Significant induction of antigen-specific serum IgY was detected in all vaccinated birds, however the magnitude and timing of SodB-specific IgY did not correlate with lower numbers of C. jejuni. Antibodies from SodB-vaccinated chickens detected the protein in the periplasm and not membrane fractions or on the bacterial surface, suggesting that the protection observed may not be strictly antibody-mediated. SodB may be useful as a constituent of vaccines for control of C. jejuni infection in broiler birds, however modest protection was observed late relative to the life of broiler birds and further studies are required to potentiate the magnitude and timing of protection. PMID:26458797

  2. Influence of relative humidity on transmission of Campylobacter jejuni in broiler chickens.

    PubMed

    Line, J E

    2006-07-01

    Horizontal transmission of Campylobacter jejuni among broiler chickens has been documented; however, the influence of RH on transmission rates is an important factor that has not been extensively studied. The purpose of our experiments was to determine the rate of C. jejuni colonization among groups of broilers raised in microbiological isolation under high (approximately 80%) and low (approximately 30%) RH conditions. Day-of-hatch chicks (n = 100 per group) were placed on wood shavings in high and low humidity-controlled pens and challenged with C. jejuni by introducing 2 seeder birds orally inoculated with C. jejuni into each group. The rate of colonization was monitored by analyzing ceca from 10 chicks from each group at d 1, 2, 3, 4, and 7. After 3 wk, the remaining chickens were removed, and 100 newly hatched chicks were placed on the contaminated litter. A second trial was conducted with the litter as the only inoculum source. Trials were repeated in this manner with the time between removing birds and placing newly hatched chicks on the litter extended to 6 h, 24 h, and 1 wk. Significant differences in Campylobacter colonization rates were observed between chickens raised under the high and low RH conditions. A delay in colonization was observed in birds raised under the low RH conditions, which increased with the increased time between removal of birds and placement of newly hatched chicks. These experiments demonstrate the importance of humidity in the transmission of Campylobacter from litter, and they could lead to practical applications to help reduce Campylobacter colonization in broilers. PMID:16830853

  3. Genome Sequence of a Clinical Isolate of Campylobacter jejuni from Thailand▿ †

    PubMed Central

    Poly, Frédéric; Read, Timothy; Tribble, David R.; Baqar, Shahida; Lorenzo, Maria; Guerry, Patricia

    2007-01-01

    Campylobacter jejuni CG8486, which belongs to the HS4 complex, was isolated from a patient with inflammatory diarrhea in Thailand. This strain caused a diarrheal disease in ferrets comparable to that caused by C. jejuni strain 81-176, but it was much less invasive for epithelial cells in vitro than 81-176. Complete genome sequencing of CG8486 revealed a 1.65-Mb genome that was very similar to the other two published genomes of clinical isolates of C. jejuni, the genomes of 81-176 and NCTC 11168, with a limited number of CG8486-specific genes mapping outside the hypervariable carbohydrate biosynthesis loci. These data suggest that the genes required for induction of inflammatory diarrhea are among the genes shared by CG8486 and 81-176 but that either major changes in the carbohydrate loci and/or more subtle changes in other genes may modulate virulence. PMID:17438034

  4. In vitro phagocytosis and intracellular survival of Campylobacter jejuni with phagocytes

    SciTech Connect

    Kiehlbauch, J.A.

    1986-01-01

    In vitro phagocytosis and intracellular survival of Campylobacter jejuni was studied using three types of mononuclear phagocytes: a J774G8 peritoneal macrophage line, resident BABL/c peritoneal macrophages and human peripheral blood monocytes. In phagocytosis assays using CFU determinations, phagocytosis increased steadily over an 8 hr time period. Results obtained using a /sup 51/Cr assay indicated no consistent significant difference between phagocytosis of C. jejuni between the three mononuclear phagocytes or PMN's and that maximum infection occurred prior to 0.5 hr and maintained throughout the 4 hr assay. Further investigation of the mechanism of attachment and entry of C. jejuni revealed this process required the expenditure of energy by the phagocyte, but was not inhibited by inhibitors of microfilament functions. In addition, phagocytosis was enhanced by the presence of 20% FCS,

  5. Risk Factors Associated with Campylobacter jejuni Infections in Curaçao, Netherlands Antilles

    PubMed Central

    Endtz, Hubert P.; van West, Hanneke; Godschalk, Peggy C. R.; de Haan, Lidewij; Halabi, Yaskara; van den Braak, Nicole; Kesztyüs, Barbara I.; Leyde, Ewald; Ott, Alewijn; Verkooyen, Roel; Price, Lawrence J.; Woodward, David L.; Rodgers, Frank G.; Ang, C. Wim; van Koningsveld, Rinske; van Belkum, Alex; Gerstenbluth, Izzy

    2003-01-01

    A steady increase in the incidence of Guillain-Barré syndrome (GBS) with a seasonal preponderance, almost exclusively related to Campylobacter jejuni, and a rise in the incidence of laboratory-confirmed Campylobacter enteritis have been reported from Curaçao, Netherlands Antilles. We therefore investigated possible risk factors associated with diarrhea due to epidemic C. jejuni. Typing by pulsed-field gel electrophoresis identified four epidemic clones which accounted for almost 60% of the infections. One hundred six cases were included in a case-control study. Infections with epidemic clones were more frequently observed in specific districts in Willemstad, the capital of Curaçao. One of these clones caused infections during the rainy season only and was associated with the presence of a deep well around the house. Two out of three GBS-related C. jejuni isolates belonged to an epidemic clone. The observations presented point toward water as a possible source of Campylobacter infections. PMID:14662945

  6. Dynamics of populations of Campylobacter jejuni in two grandparent broiler breeder farms: persistent vs. transient strains.

    PubMed

    Perez-Boto, D; Garcia-Peña, F J; Abad-Moreno, J C; Echeita, M A

    2012-09-14

    The objectives of the study were to characterize and investigate the populations of Campylobacter jejuni in two grandparent broiler breeder farms over four years. Caecal as well as farm environmental samples were obtained. Campylobacter isolates were characterized by macrorestriction profile (SmaI and KpnI-PFGE) and PCR-RFLP of the flaA gene. Susceptibility tests against seven antimicrobials were also performed. Birds were negative for Campylobacter spp. when they came to these two production farms (20 weeks), and most of the flocks remained uncolonized until they were 23 weeks old. Eighteen genotypes were characterized, with one of them (genotype 2) appearing and persisting over the study period in the two farms. In general, the strains exhibited high genetic stability, and most of them could be seen as transient in the farms, being substituted by other strains when their flock was substituted. Only one environmental sampling was positive for C. jejuni. Two different genotypes were characterized; one of them was isolated from the birds of that farm two years before. The susceptibility data point to the idea of an environmental source or reservoir of this genotype. Regarding the susceptibility of the populations, as other studies have shown, quinolone resistance (alone or combined with other resistances) was the most frequent: 68.5%. Quinolone- and multidrug-resistant strains are a matter of concern in public health. In conclusion, this survey shows the complexity of the study of the colonization of farms by C. jejuni.

  7. Development of an indirect competitive ELISA for detection of Campylobacter jejuni subsp.jejuni O:23 in foods.

    PubMed

    Hochel, I; Viochna, D; Skvor, J; Musil, M

    2004-01-01

    An indirect enzyme immunoassay for rapid detection of Campylobacter jejuni subsp. jejuni 0:23 has been developed. Optimum concentrations of immobilized cells, polyclonal chicken IgY, and rabbit anti-IgY antibody-horseradish peroxidase conjugate were 3.1 CFU/nL, 10 microg/mL, and 8 microg/mL, respectively. Under such conditions, the detection limit reached 50 CFU/microL, limit of quantification being 480 CFU/microL. By testing 5 chromogens, viz. 1,2-benzenediamine, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid), 3,3',5,5'-tetramethylbenzidine, bi(4,4'-anisidine) and 3-methyl-2-benzothiazolinone hydrazone, in horseradish peroxidase substrate, 1,2-benzenediamine or 3,3',5,5'-tetramethylbenzidine as H-donors in the enzyme substrate provided the highest ELISA sensitivity. The applied polyclonal antibody was specific for homogeneous antigen. The cross-reactions were observed only with one strain of C. sputorum subsp. sputorum (21.5 %) and with G+ bacterium Micrococcus luteus (6.1 %). Preliminary tests have been performed with a limited number of artificially contaminated food samples. No matrix effects on the ELISA sensitivity were observed. The results (by means of ELISA) were comparable with those given by both a standard cultivation method performed according to CSN ISO 10272 and commercially available Singlepath Campylobacter GLISA-Rapid Test.

  8. Transmigration route of Campylobacter jejuni across polarized intestinal epithelial cells: paracellular, transcellular or both?

    PubMed Central

    2013-01-01

    Intact intercellular junctions and cellular matrix contacts are crucial structural components for the formation and maintenance of epithelial barrier functions in humans to control the commensal flora and protect against intruding microbes. Campylobacter jejuni is one of the most important zoonotic pathogens causing food-borne gastroenteritis and potentially more severe diseases such as reactive arthritis or Guillain–Barré syndrome. Crossing the intestinal epithelial barrier and host cell invasion by C. jejuni are considered to represent the primary reasons of gut tissue damage in humans and various animal model systems including monkeys, piglets, rabbits, hamsters and ferrets. C. jejuni is also able to invade underlying tissues such as the lamina propria, can enter the bloodstream, and possibly reach distinct organs such as spleen, liver or mesenteric lymph nodes. However, the molecular mechanisms as well as major bacterial and host cell factors involved in these activities are poorly understood. Various models exist by which the pathogen can trigger its own transmigration across polarized intestinal epithelial cells in vitro, the paracellular and/or transcellular mechanism. Recent studies suggest that bacterial factors such as flagellum, serine protease HtrA and lipooligosaccharide LOS may play an active role in bacterial transmigration. Here we review our knowledge on transmigration of C. jejuni as well as some other Campylobacter species, and discuss the pros and cons for the route(s) taken to travel across polarized epithelial cell monolayers. These studies provide fresh insights into the infection strategies employed by this important pathogen. PMID:24079544

  9. Genetic Analysis of Lipooligosaccharide Core Biosynthesis in Campylobacter jejuni 81-176▿

    PubMed Central

    Kanipes, Margaret I.; Tan, Xuan; Akelaitis, Aurel; Li, Jiajun; Rockabrand, David; Guerry, Patricia; Monteiro, Mario A.

    2008-01-01

    We report isolation and characterization of Campylobacter jejuni 81-176 lgtF and galT lipooligosaccharide (LOS) core mutants. It has been suggested that the lgtF gene of C. jejuni encodes a two-domain glucosyltransferase that is responsible for the transfer of a β-1,4-glucose residue on heptosyltransferase I (Hep I) and for the transfer of a β-1,2-glucose residue on Hep II. A site-specific mutation in the lgtF gene of C. jejuni 81-176 resulted in expression of a truncated LOS, and complementation of the mutant in trans restored the core mobility to that of the wild type. Mass spectrometry and nuclear magnetic resonance of the truncated LOS confirmed the loss of two glucose residues, a β-1,4-glucose on Hep I and a β-1,2-glucose on Hep II. Mutation of another gene, galT, encoding a glycosyltransferase, which maps outside the region defined as the LOS biosynthetic locus in C. jejuni 81-176, resulted in loss of the β-(1,4)-galactose residue and all distal residues in the core. Both mutants invaded intestinal epithelial cells in vitro at levels comparable to the wild-type levels, in marked contrast to a deeper inner core waaC mutant. These studies have important implications for the role of LOS in the pathogenesis of Campylobacter-mediated infection. PMID:18156268

  10. Effects of feeding plant-derived agents on the colonization of Campylobacter jejuni in broiler chickens.

    PubMed

    Kurekci, Cemil; Al Jassim, Rafat; Hassan, Errol; Bishop-Hurley, Sharon L; Padmanabha, Jagadish; McSweeney, Christopher S

    2014-09-01

    The aim of this work was to test the potential use of plant-derived extracts and compounds to control Campylobacter jejuni in broiler chickens. Over a 7-wk feeding period, birds were fed a commercial diet with or without plant extracts (Acacia decurrens, Eremophila glabra), essential oil [lemon myrtle oil (LMO)], plant secondary compounds [terpinene-4-ol and α-tops (including α-terpineol, cineole, and terpinene-4-ol)], and the antibiotic virginiamycin. Traditional culture and real-time quantitative PCR techniques were used to enumerate the numbers of C. jejuni in chicken fecal and cecal samples. In addition, BW and feed intake were recorded weekly for the calculation of BW gain and feed conversion ratio. The mean log10 counts of C. jejuni were similar (P > 0.05) across treatments. However, significantly lower levels of fecal Campylobacter counts (P < 0.05) were recorded at d 41 for the α-tops treatment by culture methods. No differences (P > 0.05) in BW gain were obtained for dietary supplementation, except for the E. glabra extract, which had a negative impact (P < 0.001) on BW, resulting in sporadic death. Results from this study suggest that supplemental natural compounds used in the current study did not reduce the shedding of C. jejuni to desired levels.

  11. 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

    La peritonitis es un problema grave en los niños que reciben diálisis peritoneal. La bacteria Campylobacter jejuni es una causa infrecuente de peritonitis. Un niño de 10 años de edad con insuficiencia renal terminal causada por síndrome urémico hemolítico atípico ingresó a nuestro hospital con dolor abdominal y fiebre. El líquido de la diálisis peritoneal era turbio; en el examen microscópico se observaron leucocitos abundantes. Se inició tratamiento con cefepime intraperitoneal. En el cultivo del líquido peritoneal se aisló Campylobacter jejuni, por lo que se agregó claritromicina oral al tratamiento. Al finalizar el tratamiento, el resultado del cultivo del líquido peritoneal era negativo. Hasta donde sabemos, no se había informado previamente peritonitis por C. jejuni en niños. Conclusión. Si bien la peritonitis por C. jejuni es rara en los niños, debe considerarse como factor etiológico de la peritonitis.

  12. High-Voltage Electroporation of Bacteria: Genetic Transformation of Campylobacter jejuni with Plasmid DNA

    NASA Astrophysics Data System (ADS)

    Miller, Jeff F.; Dower, William J.; Tompkins, Lucy S.

    1988-02-01

    Electroporation permits the uptake of DNA by mammalian cells and plant protoplasts because it induces transient permeability of the cell membrane. We investigated the utility of high-voltage electroporation as a method for genetic transformation of intact bacterial cells by using the enteric pathogen Campylobacter jejuni as a model system. This report demonstrates that the application of high-voltage discharges to bacterial cells permits genetic transformation. Our method involves exposure of a Campylobacter cell suspension to a high-voltage exponential decay discharge (5-13 kV/cm) for a brief period of time (resistance-capacitance time constant = 2.4-26 msec) in the presence of plasmid DNA. Electrical transformation of C. jejuni results in frequencies as high as 1.2 × 106 transformants per μ g of DNA. We have investigated the effects of pulse amplitude and duration, cell growth conditions, divalent cations, and DNA concentration on the efficiency of transformation. Transformants of C. jejuni obtained by electroporation contained structurally intact plasmid molecules. In addition, evidence is presented that indicates that C. jejuni possesses DNA restriction and modification systems. The use of electroporation as a method for transforming other bacterial species and guidelines for its implementation are also discussed.

  13. Virulence spectra of typed strains of Campylobacter jejuni from different sources: a blinded in vivo study.

    PubMed

    Stewart-Tull, D E S; Coote, J G; Thompson, D H; Candlish, Denise; Wardlaw, A C; Candlish, A

    2009-05-01

    Campylobacter jejuni is a major cause of human diarrhoeal disease, but specific virulence mechanisms have not been well defined. The aims of the present blinded study were to measure and compare the in vivo properties of 40 serotyped, biotyped and genotyped C. jejuni isolates from different sources and genetic makeup. An 11-day-old chick embryo lethality assay, which measured embryo deaths and total viable bacteria over 72 h following inoculation of bacteria into the chorioallantoic membrane, revealed a spectrum of activity within the C. jejuni strains. Human and chicken isolates showed similar high virulence values for embryo deaths while the virulence of the bovine isolates was less pronounced. A one-way ANOVA comparison between the capacity of the strains to kill the chick embryos after 24 h with cytotoxicity towards cultured CaCo-2 cells was significant (P=0.025). After inoculation with a Campylobacter strain, mouse ligated ileal loops were examined histologically and revealed degrees of villous atrophy, abnormal mucosa, dilation of the lumen, congestion and blood in lumen, depending on the isolate examined. A 'total pathology score', derived for each C. jejuni strain after grading the pathology features for degree of severity, showed no apparent relationship with the source of isolation. Some relationship was found between amplified fragment length polymorphism groups and total ileal loop pathology scores, and a one-way ANOVA comparison of the mouse pathology scores against total chick embryo deaths after 72 h was significant (P=0.049). PMID:19369514

  14. Primary Isolation Strain Determines Both Phage Type and Receptors Recognised by Campylobacter jejuni Bacteriophages

    PubMed Central

    Sørensen, Martine C. Holst; Gencay, Yilmaz Emre; Birk, Tina; Baldvinsson, Signe Berg; Jäckel, Claudia; Hammerl, Jens A.; Vegge, Christina S.; Neve, Horst; Brøndsted, Lone

    2015-01-01

    In this study we isolated novel bacteriophages, infecting the zoonotic bacterium Campylobacter jejuni. These phages may be used in phage therapy of C. jejuni colonized poultry to prevent spreading of the bacteria to meat products causing disease in humans. Many C. jejuni phages have been isolated using NCTC12662 as the indicator strain, which may have biased the selection of phages. A large group of C. jejuni phages rely on the highly diverse capsular polysaccharide (CPS) for infection and recent work identified the O-methyl phosphoramidate modification (MeOPN) of CPS as a phage receptor. We therefore chose seven C. jejuni strains each expressing different CPS structures as indicator strains in a large screening for phages in samples collected from free-range poultry farms. Forty-three phages were isolated using C. jejuni NCTC12658, NCTC12662 and RM1221 as host strains and 20 distinct phages were identified based on host range analysis and genome restriction profiles. Most phages were isolated using C. jejuni strains NCTC12662 and RM1221 and interestingly phage genome size (140 kb vs. 190 kb), host range and morphological appearance correlated with the isolation strain. Thus, according to C. jejuni phage grouping, NCTC12662 and NCTC12658 selected for CP81-type phages, while RM1221 selected for CP220-type phages. Furthermore, using acapsular ∆kpsM mutants we demonstrated that phages isolated on NCTC12658 and NCTC12662 were dependent on the capsule for infection. In contrast, CP220-type phages isolated on RM1221 were unable to infect non-motile ∆motA mutants, hence requiring motility for successful infection. Hence, the primary phage isolation strain determines both phage type (CP81 or CP220) as well as receptors (CPS or flagella) recognised by the isolated phages. PMID:25585385

  15. Biological Roles of the O-Methyl Phosphoramidate Capsule Modification in Campylobacter jejuni

    PubMed Central

    Richards, Michele R.; Fodor, Christopher; Ashmus, Roger A.; Stahl, Martin; Karlyshev, Andrey V.; Wren, Brendan W.; Stintzi, Alain; Miller, William G.; Lowary, Todd L.; Szymanski, Christine M.

    2014-01-01

    Campylobacter jejuni is a major cause of bacterial gastroenteritis worldwide, and the capsular polysaccharide (CPS) of this organism is required for persistence and disease. C. jejuni produces over 47 different capsular structures, including a unique O-methyl phosphoramidate (MeOPN) modification present on most C. jejuni isolates. Although the MeOPN structure is rare in nature it has structural similarity to some synthetic pesticides. In this study, we have demonstrated, by whole genome comparisons and high resolution magic angle spinning NMR, that MeOPN modifications are common to several Campylobacter species. Using MeOPN biosynthesis and transferase mutants generated in C. jejuni strain 81–176, we observed that loss of MeOPN from the cell surface correlated with increased invasion of Caco-2 epithelial cells and reduced resistance to killing by human serum. In C. jejuni, the observed serum mediated killing was determined to result primarily from activation of the classical complement pathway. The C. jejuni MeOPN transferase mutant showed similar levels of colonization relative to the wild-type in chickens, but showed a five-fold drop in colonization when co-infected with the wild-type in piglets. In Galleria mellonella waxmoth larvae, the MeOPN transferase mutant was able to kill the insects at wild-type levels. Furthermore, injection of the larvae with MeOPN-linked monosaccharides or CPS purified from the wild-type strain did not result in larval killing, indicating that MeOPN does not have inherent insecticidal activity. PMID:24498018

  16. Passive immunization to reduce Campylobacter jejuni colonization and transmission in broiler chickens.

    PubMed

    Hermans, David; Van Steendam, Katleen; Verbrugghe, Elin; Verlinden, Marc; Martel, An; Seliwiorstow, Tomasz; Heyndrickx, Marc; Haesebrouck, Freddy; De Zutter, Lieven; Deforce, Dieter; Pasmans, Frank

    2014-01-01

    Campylobacter jejuni is the most common cause of bacterium-mediated diarrheal disease in humans worldwide. Poultry products are considered the most important source of C. jejuni infections in humans but to date no effective strategy exists to eradicate this zoonotic pathogen from poultry production. Here, the potential use of passive immunization to reduce Campylobacter colonization in broiler chicks was examined. For this purpose, laying hens were immunized with either a whole-cell lysate or the hydrophobic protein fraction of C. jejuni and their eggs were collected. In vitro tests validated the induction of specific ImmunoglobulinY (IgY) against C. jejuni in the immunized hens' egg yolks, in particular. In seeder experiments, preventive administration of hyperimmune egg yolk significantly (P < 0.01) reduced bacterial counts of seeder animals three days after oral inoculation with approximately 104 cfu C. jejuni, compared with control birds. Moreover, transmission to non-seeder birds was dramatically reduced (hydrophobic protein fraction) or even completely prevented (whole-cell lysate). Purified IgY promoted bacterial binding to chicken intestinal mucus, suggesting enhanced mucosal clearance in vivo. Western blot analysis in combination with mass spectrometry after two-dimensional gel-electrophoresis revealed immunodominant antigens of C. jejuni that are involved in a variety of cell functions, including chemotaxis and adhesion. Some of these (AtpA, EF-Tu, GroEL and CtpA) are highly conserved proteins and could be promising targets for the development of subunit vaccines. PMID:24589217

  17. Passive immunization to reduce Campylobacter jejuni colonization and transmission in broiler chickens

    PubMed Central

    2014-01-01

    Campylobacter jejuni is the most common cause of bacterium-mediated diarrheal disease in humans worldwide. Poultry products are considered the most important source of C. jejuni infections in humans but to date no effective strategy exists to eradicate this zoonotic pathogen from poultry production. Here, the potential use of passive immunization to reduce Campylobacter colonization in broiler chicks was examined. For this purpose, laying hens were immunized with either a whole-cell lysate or the hydrophobic protein fraction of C. jejuni and their eggs were collected. In vitro tests validated the induction of specific ImmunoglobulinY (IgY) against C. jejuni in the immunized hens’ egg yolks, in particular. In seeder experiments, preventive administration of hyperimmune egg yolk significantly (P < 0.01) reduced bacterial counts of seeder animals three days after oral inoculation with approximately 104 cfu C. jejuni, compared with control birds. Moreover, transmission to non-seeder birds was dramatically reduced (hydrophobic protein fraction) or even completely prevented (whole-cell lysate). Purified IgY promoted bacterial binding to chicken intestinal mucus, suggesting enhanced mucosal clearance in vivo. Western blot analysis in combination with mass spectrometry after two-dimensional gel-electrophoresis revealed immunodominant antigens of C. jejuni that are involved in a variety of cell functions, including chemotaxis and adhesion. Some of these (AtpA, EF-Tu, GroEL and CtpA) are highly conserved proteins and could be promising targets for the development of subunit vaccines. PMID:24589217

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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,...

  19. Mechanisms underlying zoonotic success of Campylobacter jejuni: the CprRS two-component regulatory system influences essential processes, biofilm formation, and pathogenesis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Mechanisms underlying zoonotic success of Campylobacter jejuni: the CprRS two-component regulatory system influences essential processes, biofilm formation, and pathogenesis Campylobacter jejuni is a leading cause of food- and waterbourne bacterial gastroenteritis in the developed world. Although il...

  20. Direct quantification of Campylobacter jejuni and Campylobacter lanienae in feces of cattle by real-time quantitative PCR.

    PubMed

    Inglis, G Douglas; Kalischuk, Lisa D

    2004-04-01

    Campylobacter species are fastidious to culture, and the ability to directly quantify biomass in microbiologically complex substrates using real-time quantitative (RTQ) PCR may enhance our understanding of their biology and facilitate the development of efficacious mitigation strategies. This study reports the use of nested RTQ-PCR to directly quantify Campylobacter jejuni and Campylobacter lanienae in cattle feces. For C. jejuni, the single-copy mapA gene was selected. For C. lanienae, the three-copy 16S rRNA gene was targeted. RTQ-PCR primers were tested alone or they were nested with species-specific primers, and amplification products were detected using the intercalating dye SYBR Green. Nesting did not increase the specificity or sensitivity of C. jejuni quantification, and the limit of quantification was 19 to 25 genome copies ( approximately 3 x 10(3) CFU/g of feces). In contrast, nested RTQ-PCR was necessary to confer specificity on C. lanienae by targeting the 16S rRNA gene. The limit of quantification was 1.8 genome copies ( approximately 250 CFU/g of feces), and there was no discernible difference between the two C. lanienae secondary primer sets evaluated. Detection and quantification of C. jejuni in naturally infested cattle feces by RTQ-PCR were comparable to the results of culture-based methods. In contrast, culturing did not detect C. lanienae in 6 of 10 fecal samples positive for the bacterium and substantially underestimated cell densities relative to nested RTQ-PCR. The results of this study illustrate that RTQ-PCR can be used to directly quantify campylobacters, including very fastidious species, in a microbiologically and chemically complex substrate. Furthermore, targeting of a multicopy universal gene provided highly sensitive quantification of C. lanienae, but nested RTQ-PCR was necessary to confer specificity. This method will facilitate subsequent studies to elucidate the impact of this group of bacteria within the gastrointestinal tracts

  1. Is the Campylobacter jejuni secretory protein Cj0069 a suitable antigen for serodiagnostics?

    PubMed

    Corso, J; Lugert, R; Groß, U; Zautner, A E

    2011-03-01

    Campylobacter spp. is the most common bacterial pathogen of gastroenteritis worldwide. Poultry is the main reservoir and consequently the main origin of infections for humans. As a consequence of a primary Campylobacter infection which typically manifests as diarrhea, there is an increased risk to suffer from post-infectious complications such as reactive arthritis, neuropathia, myositis or a Guillain-Barré Syndrome. Usually the verification of acute campylobacteriosis is made by stool culture. In contrast, post-infectious complications can be diagnosed by serological assays. Since most of them are based on whole cell lysates, an insufficient specificity results from cross-reactions between related species. Therefore, the use of recombinant antigens becomes more and more favorable. Campylobacter is able to secrete a number of proteins, which are amongst others necessary for cell invasion and therefore play a crucial role for virulence. One of these, Cj0069, has a similar specificity and sensitivity in the detection of anti-Campylobacter jejuni IgG compared to the well-established antigens OMP18 and P39. This makes it a suitable antigen for diagnosing C. jejuni post-infectious complications.

  2. Identification of nine sequence types of the 16S rRNA genes of Campylobacter jejuni subsp. jejuni isolated from broilers

    PubMed Central

    Hansson, Ingrid; Persson, Marianne; Svensson, Linda; Engvall, Eva Olsson; Johansson, Karl-Erik

    2008-01-01

    Background Campylobacter is the most commonly reported bacterial cause of enteritis in humans in the EU Member States and other industrialized countries. One significant source of infection is broilers and consumption of undercooked broiler meat. Campylobacter jejuni is the Campylobacter sp. predominantly found in infected humans and colonized broilers. Sequence analysis of the 16S rRNA gene is very useful for identification of bacteria to genus and species level. The objectives in this study were to determine the degree of intraspecific variation in the 16S rRNA genes of C. jejuni and C. coli and to determine whether the 16S rRNA sequence types correlated with genotypes generated by PFGE analysis of SmaI restricted genomic DNA of the strains. Methods The 16S rRNA genes of 45 strains of C. jejuni and two C. coli strains isolated from broilers were sequenced and compared with 16S rRNA sequences retrieved from the Ribosomal Database Project or GenBank. The strains were also genotyped by PFGE after digestion with SmaI. Results Sequence analyses of the 16S rRNA genes revealed nine sequence types of the Campylobacter strains and the similarities between the different sequence types were in the range 99.6–99.9%. The number of nucleotide substitutions varied between one and six among the nine 16S rRNA sequence types. One of the nine 16S rRNA sequence profiles was common to 12 of the strains from our study and two of these were identified as Campylobacter coli by PCR/REA. The other 10 strains were identified as Campylobacter jejuni. Five of the nine sequence types were also found among the Campylobacter sequences deposited in GenBank. The three 16S rRNA genes in the analysed strains were identical within each individual strain for all 47 strains. Conclusion C. jejuni and C. coli seem to lack polymorphisms in their 16S rRNA gene, but phylogenetic analysis based on 16S rRNA sequences was not always sufficient for differentiation between C. jejuni and C. coli. The strains

  3. Detection of flagellar antigen of Campylobacter jejuni and Campylobacter coli in canine faeces with an enzyme-linked immunosorbent assay (ELISA)--new prospects for diagnosis.

    PubMed

    Monfort, J D; Bech-Nielsen, S; Stills, H F

    1994-01-01

    A new diagnostic procedure was developed to detect the flagellar antigen of Campylobacter jejuni and Campylobacter coli in canine faecal specimens and was tested on faecal samples from random-source dogs obtained from the local dog pound. Extraction of acid-soluble proteins was performed on faecal specimens and the extracted material was evaluated using species-specific monoclonal antibodies in an enzyme-linked immunosorbent assay. The assay detected all C. jejuni or C. coli infected specimens compared with direct selective faecal culture. One of 18 faecal specimens culture-negative for C. jejuni was identified as positive by the assay, i.e. a false positive rate of 1 of 18 (5.6%) and a corresponding specificity of 94.4%. These results suggest that the screening procedure developed to detect flagellar antigens of C. jejuni and C. coli in canine faecal samples should be further investigated as a diagnostic alternative to culture.

  4. Screening for lactic acid bacteria capable of inhibiting Campylobacter jejuni in in vitro simulations of the broiler chicken caecal environment.

    PubMed

    Robyn, J; Rasschaert, G; Messens, W; Pasmans, F; Heyndrickx, M

    2012-12-01

    Thermotolerant Campylobacter spp., specifically Campylobacter jejuni and Campylobacter coli, are the most common bacterial causes of human gastroenteritis in developed countries. Consumption of improperly prepared poultry products and cross contamination are among the main causes of human campylobacteriosis. The aim of this study was to identify lactic acid bacterial (LAB) strains capable of inhibiting C. jejuni growth in initial in vitro trials ('spot-on-lawn' method), as well as in batch fermentation studies mimicking the broiler caecal environment. These experiments served as an indication for using these strains to decrease the capability of Campylobacter to colonise and grow in the chicken caeca during primary production, with the aim of reducing the number of human campylobacteriosis cases. A total of 1,150 LAB strains were screened for anti-Campylobacter activity. Six strains were selected: members of the species Lactobacillus reuteri, Lactobacillus agilis, Lactobacillus helveticus, Lactobacillus salivarius, Enterococcus faecalis and Enterococcus faecium. After treatment with catalase, proteinase K and a-chymotrypsin, anti-Campylobacter activity of cell-free culture supernatant fluid (CSF) for all six strains was retained, which indicated that activity was probably not exerted by bacteriocin production. Based on the activity found in CSF, the compounds produced by the selected strains are secreted and do not require presence of live bacterial producer cells for activity. During initial in vitro fermentation experiments, the E. faecalis strain exhibited the highest inhibitory activity for C. jejuni and was selected for further fermentation experiments. In these experiments we tested for therapeutic or protective effects of the E. faecalis strain against C. jejuni MB 4185 infection under simulated broiler caecal growth conditions. The best inhibition results were obtained when E. faecalis was inoculated before the C. jejuni strain, lowering C. jejuni counts at

  5. [The study of influence of stresses on virulence genes expression in foodborne pathogens Campylobacter jejuni].

    PubMed

    Efimochkina, N R; Bykova, I B; Markova, Yu M; Korotkevich, Yu V; Sheveleva, S A

    2016-01-01

    The study of the responses to cold exposure in Campylobacterjejuni (C. jejuni)--one of the most common foodborne pathogens is important for elucidating the mechanisms of acquisition of products contaminated with campylobacter, hazardous properties. These data are also necessary to create effective systems of microbiological controls at all stages of production and storage of food. 5 pairs of oligonucleotide primers were selected for detecting of genes cadF, cdtB, ciaB, flaA, iamA, encoding the main factors of pathogenicity of foodborne pathogens Campylobacter jejuni--adhesion and invasion of epithelial cells, production of CDT-toxin and mobility. To quantify the expression levels of target genes of C. jejuni a comparative method of determining the amount of amplification products of genes encoding pathogenicity factors of Campylobacter spp. has been developed using real-time PCR with intercalating dyes. To calculate and quantify gene expression the mathematical models have been obtained that allow extrapolation of threshold cycles of amplification to the initial number of copies of RNA/DNA in the tested samples. It has been established that exposure of C. jejuni at low temperatures +4 degrees C did not lead to increased levels of expression of genes cdtB and ciaB. However, in the populations of C. jejuni subjected to freezing, followed by incubation at optimum for the pathogen temperature of +42 degrees C, the increase in expression of mRNA encoding protein subunit B of CDT-toxin and antigenic marker of invasion took place. The number of copies of RNA in C. jejuni after stress exposure increased by 1.14-2.6 lg in comparison with intact cultures. CdtB and ciaB gene expression in C. jejuni can serve as an indicator of cell response to stress and helps to restore the functions of the bacterial cells after the termination of cold exposure and return of the pathogen in conditions favourable to the realization of its pathogenic potential. PMID:27228703

  6. [The study of influence of stresses on virulence genes expression in foodborne pathogens Campylobacter jejuni].

    PubMed

    Efimochkina, N R; Bykova, I B; Markova, Yu M; Korotkevich, Yu V; Sheveleva, S A

    2016-01-01

    The study of the responses to cold exposure in Campylobacterjejuni (C. jejuni)--one of the most common foodborne pathogens is important for elucidating the mechanisms of acquisition of products contaminated with campylobacter, hazardous properties. These data are also necessary to create effective systems of microbiological controls at all stages of production and storage of food. 5 pairs of oligonucleotide primers were selected for detecting of genes cadF, cdtB, ciaB, flaA, iamA, encoding the main factors of pathogenicity of foodborne pathogens Campylobacter jejuni--adhesion and invasion of epithelial cells, production of CDT-toxin and mobility. To quantify the expression levels of target genes of C. jejuni a comparative method of determining the amount of amplification products of genes encoding pathogenicity factors of Campylobacter spp. has been developed using real-time PCR with intercalating dyes. To calculate and quantify gene expression the mathematical models have been obtained that allow extrapolation of threshold cycles of amplification to the initial number of copies of RNA/DNA in the tested samples. It has been established that exposure of C. jejuni at low temperatures +4 degrees C did not lead to increased levels of expression of genes cdtB and ciaB. However, in the populations of C. jejuni subjected to freezing, followed by incubation at optimum for the pathogen temperature of +42 degrees C, the increase in expression of mRNA encoding protein subunit B of CDT-toxin and antigenic marker of invasion took place. The number of copies of RNA in C. jejuni after stress exposure increased by 1.14-2.6 lg in comparison with intact cultures. CdtB and ciaB gene expression in C. jejuni can serve as an indicator of cell response to stress and helps to restore the functions of the bacterial cells after the termination of cold exposure and return of the pathogen in conditions favourable to the realization of its pathogenic potential.

  7. 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

  8. HtrA chaperone activity contributes to host cell binding in Campylobacter jejuni

    PubMed Central

    2011-01-01

    Background Acute gastroenteritis caused by the food-borne pathogen Campylobacter jejuni is associated with attachment of bacteria to the intestinal epithelium and subsequent invasion of epithelial cells. In C. jejuni, the periplasmic protein HtrA is required for efficient binding to epithelial cells. HtrA has both protease and chaperone activity, and is important for virulence of several bacterial pathogens. Results The aim of this study was to determine the role of the dual activities of HtrA in host cell interaction of C. jejuni by comparing an htrA mutant lacking protease activity, but retaining chaperone activity, with a ΔhtrA mutant and the wild type strain. Binding of C. jejuni to both epithelial cells and macrophages was facilitated mainly by HtrA chaperone activity that may be involved in folding of outer membrane adhesins. In contrast, HtrA protease activity played only a minor role in interaction with host cells. Conclusion We show that HtrA protease and chaperone activities contribute differently to C. jejuni's interaction with mammalian host cells, with the chaperone activity playing the major role in host cell binding. PMID:21939552

  9. Messenger RNA expression of chicken CLOCK gene in the response to Campylobacter jejuni inoculation.

    PubMed

    Liu, Xiaoyi; Liu, Liying; Zhang, Maozhi; Yang, Ning; Qi, Yukai; Sun, Yu; Li, Xianyao

    2015-09-01

    Campylobacter jejuni (C. jejuni) is a leading cause of human bacterial gastroenteritis worldwide. Previous research has shown that circadian rhythm plays a critical role in host response to C. jejuni colonization. The CLOCK gene is one of the core genes regulating circadian rhythms and shows significant expression on 7 d post-C. jejuni inoculation. The objective of this study was to investigate temporal and spatial expression of chicken CLOCK gene post-C. jejuni inoculation. Cecal and splenic RNA were isolated from 2 distinct chicken breeds and used to compare the mRNA expression of CLOCK gene between inoculated and noninoculated chickens within each breed and between breeds within each of inoculated and noninoculated groups. Our results showed that the CLOCK gene was significantly down-regulated at 20 h postinoculation (hpi) in cecum and spleen in Jiningbairi chicken. CLOCK gene was significantly down-regulated at 4 and 16 hpi and up-regulated at 8 hpi in cecum and spleen in specific pathogen free white leghorn noninoculated chicken. The findings suggested that expression of CLOCK gene was significantly changed post C. jejuin inoculation. This change was affected by genetic background, tissue, and time points postinoculation.

  10. Campylobacter jejuni as a cause of canine abortions in the United States.

    PubMed

    Sahin, Orhan; Burrough, Eric R; Pavlovic, Nada; Frana, Tim S; Madson, Darin M; Zhang, Qijing

    2014-09-01

    Although Campylobacter jejuni is a common cause of ruminant abortion with great economic impact, the organism has rarely been implicated in canine pregnancy loss, with only 2 documented cases to date. In the current report, 2 cases of perinatal death in adult female Bulldogs associated with C. jejuni infection of fetoplacental organs are described. Fetuses and placentas were received at the Iowa State University Veterinary Diagnostic Laboratory (Ames, Iowa) from 3 puppies that died soon after the birth (case 1) and from an aborted fetus (case 2). Microscopic examination of tissues was generally unremarkable; however, multifocal hemorrhage and infiltrates of macrophages and neutrophils were observed in placental sections from the first case (case 1), and low to moderate numbers of degenerate neutrophils were apparent within multifocal alveoli in the fetal lung in the second case (case 2). Ancillary diagnostics for common infectious causes of reproductive failure in dogs were negative. However, C. jejuni was isolated from the submitted placentas in high numbers in both cases as well as from the fetal lungs and livers. Genotyping of the abortion isolates indicated that the isolates were distinct from each other as well as from selected canine enteric C. jejuni isolates included herein for comparison. Both abortion strains were sensitive to all 9 antimicrobials tested, except the isolate from case 2, which displayed resistance to tetracycline. These findings provide convincing evidence for the inclusion of C. jejuni culture in routine diagnostic testing for causes of canine pregnancy loss.

  11. Survival of Campylobacter jejuni and Salmonella enterica Typhimurium in vacuum-packed, moisture-enhanced pork.

    PubMed

    Wen, Xuesong; Dickson, James S

    2012-03-01

    The abilities of Campylobacter jejuni and Salmonella enterica Typhimurium to survive in vacuum-packaged, moisture-enhanced pork stored at 4 or 10°C were examined. Pork loins were surface inoculated with either C. jejuni or Salmonella Typhimurium and then moisture enhanced to a target of 10 or 20%. The enhanced pork loins were sliced 1 cm thick and vacuum packaged. A pork loin without moisture enhancement was sliced and vacuum packaged as a control. Samples were collected, plated, and the numbers of surviving organisms were determined periodically during storage at 4 and 10°C. The numbers of C. jejuni or Salmonella Typhimurium in samples with different moisture enhancement levels were similar (P > 0.05). No significant differences (P > 0.05) in C. jejuni counts were observed between samples at 10°C and those at 4°C. In contrast, the numbers of Salmonella Typhimurium in samples at 10°C had significantly (P < 0.05) increased (0.41 log CFU/g) from those at the refrigerated temperature of 4°C. Vacuum storage at 4 and 10°C for 28 days did not result in dramatic reductions in the mean numbers of C. jejuni and Salmonella Typhimurium. Our findings indicate that vacuum packaging under chilled conditions will not add substantially to safety for moisture-enhanced pork. Strict hygienic practices or the implementation of decontamination technologies is recommended.

  12. Exposure of Campylobacter jejuni to 6 degrees C: effects on heat resistance and electron transport activity.

    PubMed

    Hughes, Rebecca-Ayme; Cogan, Tristan; Humphrey, Tom

    2010-04-01

    Human infection with Campylobacter jejuni is frequently associated with the consumption of foods, especially chicken meat, which have been exposed to a range of temperatures during processing, storage, and cooking. Despite the public health importance of C. jejuni, little is known about the effects of cold exposure (refrigeration) on the subsequent ability of this pathogen to survive heat challenge. This work examined the effect of rapid exposure to 6 degrees C for 24 h on the heat resistance at 52 degrees C of 19 C. jejuni strains originally isolated from various sources. The resulting death curves were analyzed with the Weibull model. Unlike cold-exposed cells of Escherichia coli and Salmonella, which have been reported to show significant increased sensitivity to heat, such exposure had only a marginal effect on heat resistance of the C. jejuni strains in this study. A possible explanation for this effect is that rapid chilling renders C. jejuni cells unable to adapt to reduced temperatures in an active manner. This hypothesis is supported by the observation that exposure to 6 degrees C for 24 h resulted in a significant and marked reduction in electron transport system activity when compared with controls at 37 degrees C.

  13. Campylobacter jejuni actively invades the amoeba Acanthamoeba polyphaga and survives within non digestive vacuoles.

    PubMed

    Olofsson, Jenny; Axelsson-Olsson, Diana; Brudin, Lars; Olsen, Björn; Ellström, Patrik

    2013-01-01

    The Gram-negative bacterium Campylobacter jejuni is able to enter, survive and multiply within the free living amoeba Acanthamoeba polyphaga, but the molecular mechanisms behind these events are still unclear. We have studied the uptake and intracellular trafficking of viable and heat killed bacterial cells of the C. jejuni strain 81-176 in A. polyphaga. We found that viable bacteria associated with a substantially higher proportion of Acanthamoeba trophozoites than heat killed bacteria. Furthermore, the kinetics of internalization, the total number of internalized bacteria as well as the intracellular localization of internalized C. jejuni were dramatically influenced by bacterial viability. Viable bacteria were internalized at a high rate already after 1 h of co-incubation and were observed in small vacuoles tightly surrounding the bacteria. In contrast, internalization of heat killed C. jejuni was low at early time points and did not peak until 96 h. These cells were gathered in large spacious vacuoles that were part of the degradative pathway as determined by the uptake of fluorescently labeled dextran. The amount of heat killed bacteria internalized by A. polyphaga did never reach the maximal amount of internalized viable bacteria. These results suggest that the uptake and intracellular survival of C. jejuni in A. polyphaga is bacterially induced. PMID:24223169

  14. Campylobacter jejuni sequence types show remarkable spatial and temporal stability in Blackbirds

    PubMed Central

    Griekspoor, Petra; Hansbro, Philip M.; Waldenström, Jonas; Olsen, Björn

    2015-01-01

    Background The zoonotic bacterium Campylobacter jejuni has a broad host range but is especially associated with birds, both domestic and wild. Earlier studies have indicated thrushes of the genus Turdus in Europe to be frequently colonized with C. jejuni, and predominately with host-associated specific genotypes. The European Blackbird Turdus merula has a large distribution in Europe, including some oceanic islands, and was also introduced to Australia by European immigrants in the 1850s. Methods The host specificity and temporal stability of European Blackbird C. jejuni was investigated with multilocus sequence typing in a set of isolates collected from Sweden, Australia, and The Azores. Results Remarkably, we found that the Swedish, Australian, and Azorean isolates were genetically highly similar, despite extensive spatial and temporal isolation. This indicates adaptation, exquisite specificity, and stability in time for European Blackbirds, which is in sharp contrast with the high levels of recombination and mutation found in poultry-related C. jejuni genotypes. Conclusion The maintenance of host-specific signals in spatially and temporally separated C. jejuni populations suggests the existence of strong purifying selection for this bacterium in European Blackbirds. PMID:26634844

  15. Matrix Metalloproteinase-2 Mediates Intestinal Immunopathogenesis in Campylobacter Jejuni-Infected Infant Mice

    PubMed Central

    Alutis, Marie E.; Grundmann, Ursula; Hagen, Ulrike; Fischer, André; Kühl, Anja A.; Göbel, Ulf B.; Bereswill, Stefan; Heimesaat, Markus M.

    2015-01-01

    Increased levels of the matrix metalloproteinases (MMPs)-2 and -9 (also referred to gelatinase-A and -B, respectively) can be detected in the inflamed gut. We have recently shown that synthetic gelatinase blockage reduces colonic apoptosis and pro-inflammatory immune responses following murine Campylobacter (C.) jejuni infection. In order to dissect whether MMP-2 and/or MMP-9 is involved in mediating C. jejuni-induced immune responses, infant MMP-2–/–, MMP-9–/–, and wildtype (WT) mice were perorally infected with the C. jejuni strain B2 immediately after weaning. Whereas, at day 2 postinfection (p.i.), fecal C. jejuni B2 loads were comparable in mice of either genotype, mice expelled the pathogen from the intestinal tract until day 4 p.i. Six days p.i., colonic MMP-2 but not MMP-9 mRNA was upregulated in WT mice. Remarkably, infected MMP-2–/– mice exhibited less frequent abundance of blood in feces, less distinct colonic histopathology and apoptosis, lower numbers of effector as well as innate and adaptive immune cells within the colonic mucosa, and higher colonic IL-22 mRNA levels as compared to infected WT mice. In conclusion, these results point towards an important role of MMP-2 in mediating C. jejuni-induced intestinal immunopathogenesis. PMID:26495129

  16. Genotypes and Antibiotic Resistances of Campylobacter jejuni Isolates from Cattle and Pigeons in Dairy Farms

    PubMed Central

    Bianchini, Valentina; Luini, Mario; Borella, Laura; Parisi, Antonio; Jonas, Romie; Kittl, Sonja; Kuhnert, Peter

    2014-01-01

    Campylobacter jejuni is the most common food-borne zoonotic pathogen causing human gastroenteritis worldwide and has assumed more importance in Italy following the increased consumption of raw milk. Our objectives were to get an overview of genotypes and antibiotic resistances in C. jejuni isolated from milk, cattle feces, and pigeons in dairy herds of Northern Italy. flaB-typing was applied to 78 C. jejuni isolates, previously characterized by Multi-Locus Sequence Typing, and genotypic resistances towards macrolides and quinolones based on point mutations in the 23S rRNA and gyrA genes, respectively, were determined. flaB-typing revealed 22 different types with one of them being novel and was useful to further differentiate strains with an identical Sequence Type (ST) and to identify a pigeon-specific clone. Macrolide resistance was not found, while quinolone resistance was detected in 23.3% of isolates. A relationship between specific genotypes and antibiotic resistance was observed, but was only significant for the Clonal Complex 206. Our data confirm that pigeons do not play a role in the spread of C. jejuni among cattle and they are not responsible for milk contamination. A relevant number of bulk milk samples were contaminated by C. jejuni resistant to quinolones, representing a possible source of human resistant strains. PMID:25026083

  17. Reconstitution of a Functional Toll-like Receptor 5 Binding Site in Campylobacter jejuni Flagellin*

    PubMed Central

    de Zoete, Marcel R.; Keestra, A. Marijke; Wagenaar, Jaap A.; van Putten, Jos P. M.

    2010-01-01

    Bacterial flagellin is important for intestinal immune homeostasis. Flagellins from most species activate Toll-like receptor 5 (TLR5). The principal bacterial food-borne pathogen Campylobacter jejuni escapes TLR5 recognition, probably due to an alternate flagellin subunit structure. We investigated the molecular basis of TLR5 evasion by aiming to reconstitute TLR5 stimulating activity in live C. jejuni. Both native glycosylated C. jejuni flagellins (FlaA and FlaB) and recombinant proteins purified from Escherichia coli failed to activate NF-κB in HEK293 cells expressing TLR5. Introduction of multiple defined regions from Salmonella flagellin into C. jejuni FlaA via a recombinatorial approach revealed three regions critical for the activation of human and mouse TLR5, including a β-hairpin structure not previously implicated in TLR5 recognition. Surprisingly, this domain was not required for the activation of chicken TLR5, indicating a selective requirement for the β-hairpin in the recognition of mammalian TLR5. Expression of the active chimeric protein in C. jejuni resulted in secreted glycosylated flagellin that induced a potent TLR5 response. Overall, our results reveal a novel structural requirement for TLR5 recognition of bacterial flagellin and exclude flagellin glycosylation as an additional mechanism of bacterial evasion of the TLR5 response. PMID:20164175

  18. Reconstitution of a functional Toll-like receptor 5 binding site in Campylobacter jejuni flagellin.

    PubMed

    de Zoete, Marcel R; Keestra, A Marijke; Wagenaar, Jaap A; van Putten, Jos P M

    2010-04-16

    Bacterial flagellin is important for intestinal immune homeostasis. Flagellins from most species activate Toll-like receptor 5 (TLR5). The principal bacterial food-borne pathogen Campylobacter jejuni escapes TLR5 recognition, probably due to an alternate flagellin subunit structure. We investigated the molecular basis of TLR5 evasion by aiming to reconstitute TLR5 stimulating activity in live C. jejuni. Both native glycosylated C. jejuni flagellins (FlaA and FlaB) and recombinant proteins purified from Escherichia coli failed to activate NF-kappaB in HEK293 cells expressing TLR5. Introduction of multiple defined regions from Salmonella flagellin into C. jejuni FlaA via a recombinatorial approach revealed three regions critical for the activation of human and mouse TLR5, including a beta-hairpin structure not previously implicated in TLR5 recognition. Surprisingly, this domain was not required for the activation of chicken TLR5, indicating a selective requirement for the beta-hairpin in the recognition of mammalian TLR5. Expression of the active chimeric protein in C. jejuni resulted in secreted glycosylated flagellin that induced a potent TLR5 response. Overall, our results reveal a novel structural requirement for TLR5 recognition of bacterial flagellin and exclude flagellin glycosylation as an additional mechanism of bacterial evasion of the TLR5 response. PMID:20164175

  19. Occurrence and population density of Campylobacter jejuni in irrigation ponds on produce farms in the Suwannee River Watershed.

    PubMed

    Gu, Ganyu; Luo, Zhiyao; Cevallos-Cevallos, Juan M; Adams, Paige; Vellidis, George; Wright, Anita; van Bruggen, Ariena H C

    2013-05-01

    Campylobacter spp., especially Campylobacter jejuni, are common causal agents of gastroenteritis globally. Poultry, contaminated water, and fresh produce are considered to be the main sources for infection by this pathogen. In this study, occurrence and population density of C. jejuni from vegetable irrigation ponds in the Suwannee River watershed were investigated and the relationship to environmental factors was analyzed. Two water samples were collected from each of 10 ponds every month from January 2011 to February 2012. Campylobacter jejuni was detected by quantitative real-time PCR. Nine of the 10 ponds were positive for C. jejuni some of the time with an overall prevalence of 19.3%. The highest counts were obtained in spring 2011. Oxidation-reduction potential and total nitrogen concentration were positively correlated (P < 0.05) with mean population and occurrence of C. jejuni, while temperature and dissolved oxygen percent saturation (DO%) were negatively correlated with mean population (P < 0.05). Presence of this pathogen was related to bacterial community composition. No correlations were found between C. jejuni and fecal indicators. Increasing DO% of irrigation water and limiting nitrogen pollution in the ponds are suggested to reduce the contamination risk of C. jejuni in a major fruit and vegetable growing area.

  20. Gene expression profiling of the local cecal response of genetic chicken lines that differ in their susceptibility to Campylobacter jejuni colonization

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Campylobacter jejuni (C. jejuni) is one of the most common causes of human bacterial enteritis worldwide, with poultry products being one of the main sources of contamination. The molecular mechanisms of the local host response to C. jejuni infection in chickens have not been well characterized. P...

  1. Synthesis and immunodetection of 6-O-methyl-phosphoramidyl-α-D-galactose: a Campylobacter jejuni antigenic determinant.

    PubMed

    Jiao, Yuening; Ma, Zuchao; Ewing, Cheryl P; Guerry, Patricia; Monteiro, Mario A

    2015-12-11

    Campylobacter jejuni is a leading cause of traveler's diarrhea. Previously, we have shown that a C. jejuni capsule polysaccharide (CPS) conjugate vaccine can fully prevent C.jejuni diarrhea in non-human primates. C.jejuni CPSs are decorated with non-stoichiometric amounts of O-methyl phosphoramidate (MeOPN) units that are key serospecific markers. In the case of C.jejuni serotype complex HS23/36, the MeOPN are at positions 2 and 6 of the CPS galactose (Gal). We describe here the synthesis of the p-methoxyphenyl glycoside of MeOPN→6-α-D-Galp, and its immunodetection by antisera raised by C.jejuni CPS conjugates with MeOPN at primary positions. The synthetic approach in this work served as the foundation for a similar MeOPN→6-Gal construct used in a conjugate vaccine, whose synthesis, immunogenicity and efficacy will be described elsewhere.

  2. Examination of nanoparticle inactivation of Campylobacter jejuni biofilms using infrared and Raman spectroscopies

    PubMed Central

    Lu, Xiaonan; Weakley, Andrew T.; Aston, D. Eric; Rasco, Barbara A.; Wang, Shuo; Konkel, Michael E.

    2012-01-01

    Aims To investigate inactivation effect and mechanism of zinc oxide nanoparticles (ZnO NPs) activity against Campylobacter jejuni biofilms. Methods and Results ZnO NPs with concentrations of 0, 0.6, 1.2 and 6 mmol l−1 were employed in antimicrobial tests against C. jejuni planktonic cells and biofilms. C. jejuni sessile cells in biofilms were more resistant to a low concentration of ZnO NPs when compared to planktonic cells. The ZnO NPs penetrated the extracellular polymeric substance (EPS) without damage to the EPS and directly interacted with the sessile bacterial cells, as determined using infrared spectroscopy and scanning electron microscopy. Raman spectroscopy shows alterations in quinone structures and damage to nucleic acids following C. jejuni treatment with ZnO NPs. The mechanism of DNA damage is most likely due to the generation of reactive oxygen species (ROS). Spectroscopic based partial least squares regression (PLSR) models could predict the number of surviving sessile cell numbers within a bacterial biofilm (≥log 4 CFU, RMSEE <0.36) from Fourier transform infrared (FT-IR) spectral measurements. Conclusions ZnO NPs were found to have antimicrobial activity against C. jejuni biofilms. ZnO NPs penetrated the biofilm EPS within 1 hr without damaging it and interacted directly with sessile cells in biofilms. Alterations in the DNA/RNA bases, which are due to the generation of ROS, appear to result in C. jejuni cell death. Significance and Impact of the Study ZnO NPs may offer a realistic strategy to eliminate C. jejuni biofilms in the environment. PMID:22734855

  3. Antimicrobial effects of Pseudomonas aeruginosa on survivability and recovery of Campylobacter jejuni on poultry products.

    PubMed

    Davis, M A; Conner, D E

    2007-04-01

    Three types of poultry products representing differences in skin coverage were tested to determine the ability of Pseudomonas aeruginosa to inhibit growth of Campylobacter jejuni. Processed ready-to-cook poultry carcasses were obtained from the Poultry Research Unit at Auburn University and were not subjected to any treatment to reduce or eliminate the native microflora on the carcasses. Carcasses were cut into wing sections (drumette, flat, tip), split breast pieces (with and without bone), and boneless, skinless breast pieces. Equal numbers of the 3 product types were subjected to 1 of 6 treatments: 1) uninoculated, 2) C. jejuni only, 3) P. aeruginosa type 1 only, 4) P. aeruginosa type 2 only, 5) C. jejuni + P. aeruginosa type 1, or 6) C. jejuni + P. aeruginosa type 2. Products were inoculated at 10(4) to 10(5) cfu. Postinoculation, equal numbers of product type were also subjected to the following: 1) aerobic or vacuum packaging, 2) storage temperature of 4 or 10 degrees C, and 3) storage of 0, 1, 2, 3, or 4 d. Products were sampled after storage duration to determine the population of C. jejuni and P. aeruginosa. Individual pieces were rinsed with 50 mL of buffered peptone water. The recovered rinse was used to make appropriate dilutions and spiral plated onto Campy-Cefex and Pseudomonas P agars. Campy-Cefex plates were incubated microaerophilically at 42 degrees C for 48 h, whereas Pseudomonas P plates were incubated aerobically at 37 degrees C for 24 to 48 h. Random suspect colonies on Campy-Cefex plates were confirmed by cell morphology when viewed under microscopic examination. Suspect colonies on Pseudomonas P plates produced a blue color in the medium indicative of glycerol reduction. At both 4 and 10 degrees C, neither type of P. aeruginosa inhibited the growth or survival of C. jejuni compared to plates that were inoculated with C. jejuni only. PMID:17369550

  4. Quinolone and macrolide resistance in Campylobacter jejuni and C. coli: resistance mechanisms and trends in human isolates.

    PubMed Central

    Engberg, J.; Aarestrup, F. M.; Taylor, D. E.; Gerner-Smidt, P.; Nachamkin, I.

    2001-01-01

    The incidence of human Campylobacter jejuni and C. coli infections has increased markedly in many parts of the world in the last decade as has the number of quinolone-resistant and, to a lesser extent, macrolide-resistant Campylobacter strains causing infections. We review macrolide and quinolone resistance in Campylobacter and track resistance trends in human clinical isolates in relation to use of these agents in food animals. Susceptibility data suggest that erythromycin and other macrolides should remain the drugs of choice in most regions, with systematic surveillance and control measures maintained, but fluoroquinolones may now be of limited use in the empiric treatment of Campylobacter infections in many regions. PMID:11266291

  5. 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.

  6. Phenotypic and genotypic evidence for L-fucose utilization by Campylobacter jejuni.

    PubMed

    Muraoka, Wayne T; Zhang, Qijing

    2011-03-01

    Campylobacter jejuni remains among the leading causes of bacterial food-borne illness. The current understanding of Campylobacter physiology suggests that it is asaccharolytic and is unable to catabolize exogenous carbohydrates. Contrary to this paradigm, we provide evidence for l-fucose utilization by C. jejuni. The fucose phenotype, shown in chemically defined medium, is strain specific and linked to an 11-open reading frame (ORF) plasticity region of the bacterial chromosome. By constructing a mutation in fucP (encoding a putative fucose permease), one of the genes in the plasticity region, we found that this locus is required for fucose utilization. Consistent with their function in fucose utilization, transcription of the genes in the locus is highly inducible by fucose. PCR screening revealed a broad distribution of this genetic locus in strains derived from various host species, and the presence of this locus was consistently associated with fucose utilization. Birds inoculated with the fucP mutant strain alone were colonized at a level comparable to that by the wild-type strain; however, in cocolonization experiments, the mutant was significantly outcompeted by the wild-type strain when birds were inoculated with a low dose (10⁵ CFU per bird). This advantage was not observed when birds were inoculated at a higher inoculum dose (10⁸ CFU per bird). These results demonstrated a previously undescribed substrate that supports growth of C. jejuni and identified the genetic locus associated with the utilization of this substrate. These findings substantially enhance our understanding of the metabolic repertoire of C. jejuni and the role of metabolic diversity in Campylobacter pathobiology.

  7. Commonality and biosynthesis of the O-methyl phosphoramidate capsule modification in Campylobacter jejuni.

    PubMed

    McNally, David J; Lamoureux, Marc P; Karlyshev, Andrey V; Fiori, Laura M; Li, Jianjun; Thacker, Gillian; Coleman, Russell A; Khieu, Nam H; Wren, Brendan W; Brisson, Jean-Robert; Jarrell, Harold C; Szymanski, Christine M

    2007-09-28

    In this study we investigated the commonality and biosynthesis of the O-methyl phosphoramidate (MeOPN) group found on the capsular polysaccharide (CPS) of Campylobacter jejuni. High resolution magic angle spinning NMR spectroscopy was used as a rapid, high throughput means to examine multiple isolates, analyze the cecal contents of colonized chickens, and screen a library of CPS mutants for the presence of MeOPN. Sixty eight percent of C. jejuni strains were found to express the MeOPN with a high prevalence among isolates from enteritis, Guillain Barré, and Miller-Fisher syndrome patients. In contrast, MeOPN was not observed for any of the Campylobacter coli strains examined. The MeOPN was detected on C. jejuni retrieved from cecal contents of colonized chickens demonstrating that the modification is expressed by bacteria inhabiting the avian gastrointestinal tract. In C. jejuni 11168H, the cj1415-cj1418 cluster was shown to be involved in the biosynthesis of MeOPN. Genetic complementation studies and NMR/mass spectrometric analyses of CPS from this strain also revealed that cj1421 and cj1422 encode MeOPN transferases. Cj1421 adds the MeOPN to C-3 of the beta-d-GalfNAc residue, whereas Cj1422 transfers the MeOPN to C-4 of D-glycero-alpha-L-gluco-heptopyranose. CPS produced by the 11168H strain was found to be extensively modified with variable MeOPN, methyl, ethanolamine, and N-glycerol groups. These findings establish the importance of the MeOPN as a diagnostic marker and therapeutic target for C. jejuni and set the groundwork for future studies aimed at the detailed elucidation of the MeOPN biosynthetic pathway.

  8. Characterization of Campylobacter jejuni DNA gyrase as the target of quinolones.

    PubMed

    Changkwanyeun, Ruchirada; Usui, Masaru; Kongsoi, Siriporn; Yokoyama, Kazumasa; Kim, Hyun; Suthienkul, Orasa; Changkaew, Kanjana; Nakajima, Chie; Tamura, Yutaka; Suzuki, Yasuhiko

    2015-08-01

    Quinolones have long been used as the first-line treatment for Campylobacter infections. However, an increased resistance to quinolones has raised public health concerns. The development of new quinolone-based antibiotics with high activity is critical for effective, as DNA gyrase, the target of quinolones, is an essential enzyme for bacterial growth in several mechanisms. The evaluation of antibiotic activity against Campylobacter jejuni largely relies on drug susceptibility tests, which require at least 2 days to produce results. Thus, an in vitro method for studying the activity of quinolones against the C. jejuni DNA gyrase is preferred. To identify potent quinolones, we investigated the interaction of C. jejuni DNA gyrase with a number of quinolones using recombinant subunits. The combination of purified subunits exhibited DNA supercoiling activity in an ATP dependent manner. Drug concentrations that inhibit DNA supercoiling by 50% (IC50s) of 10 different quinolones were estimated to range from 0.4 (sitafloxacin) to >100 μg/mL (nalidixic acid). Sitafloxacin showed the highest inhibitory activity, and the analysis of the quinolone structure-activity relationship demonstrated that a fluorine atom at R-6 might play the important role in the inhibitory activity against C. jejuni gyrase. Measured quinolone IC50s correlated well with minimum inhibitory concentrations (R = 0.9943). These suggest that the in vitro supercoiling inhibition assay on purified recombinant C. jejuni DNA gyrase is a useful and predictive technique to monitor the antibacterial potency of quinolones. And furthermore, these data suggested that sitafloxacin might be a good candidate for clinical trials on campylobacteriosis.

  9. Comparative population structure analysis of Campylobacter jejuni from human and poultry origin in Bangladesh.

    PubMed

    Islam, Z; van Belkum, A; Wagenaar, J A; Cody, A J; de Boer, A G; Sarker, S K; Jacobs, B C; Talukder, K A; Endtz, H P

    2014-12-01

    Campylobacter jejuni is the most important cause of antecedent infections leading to Guillain-Barré syndrome (GBS) and Miller Fisher syndrome (MFS). The objective of the present study was to define the genetic diversity, population structure, and potential role of poultry in the transmission of Campylobacter to humans in Bangladesh. We determined the population structure of C. jejuni isolated from poultry (n = 66) and patients with enteritis (n = 39) or GBS (n = 10). Lipooligosaccharide (LOS) typing showed that 50/66 (76 %) C. jejuni strains isolated from poultry could be assigned to one of five LOS locus classes (A-E). The distribution of neuropathy-associated LOS locus classes A, B, and C were 30/50 (60 %) among the typable strains isolated from poultry. The LOS locus classes A, B, and C were significantly associated with GBS and enteritis-related C. jejuni strains more than for the poultry strains [(31/38 (82 %) vs. 30/50 (60 %), p < 0.05]. Multilocus sequence typing (MLST) defined 15 sequence types (STs) and six clonal complexes (CCs) among poultry isolates, including one ST-3740 not previously documented. The most commonly identified type, ST-5 (13/66), in chicken was seen only once among human isolates (1/49) (p < 0.001). Amplified fragment length polymorphism (AFLP) revealed three major clusters (A, B, and C) among C. jejuni isolated from humans and poultry. There seems to be a lack of overlap between the major human and chicken clones, which suggests that there may be additional sources for campylobacteriosis other than poultry in Bangladesh.

  10. Comparative population structure analysis of Campylobacter jejuni from human and poultry origin in Bangladesh.

    PubMed

    Islam, Z; van Belkum, A; Wagenaar, J A; Cody, A J; de Boer, A G; Sarker, S K; Jacobs, B C; Talukder, K A; Endtz, H P

    2014-12-01

    Campylobacter jejuni is the most important cause of antecedent infections leading to Guillain-Barré syndrome (GBS) and Miller Fisher syndrome (MFS). The objective of the present study was to define the genetic diversity, population structure, and potential role of poultry in the transmission of Campylobacter to humans in Bangladesh. We determined the population structure of C. jejuni isolated from poultry (n = 66) and patients with enteritis (n = 39) or GBS (n = 10). Lipooligosaccharide (LOS) typing showed that 50/66 (76 %) C. jejuni strains isolated from poultry could be assigned to one of five LOS locus classes (A-E). The distribution of neuropathy-associated LOS locus classes A, B, and C were 30/50 (60 %) among the typable strains isolated from poultry. The LOS locus classes A, B, and C were significantly associated with GBS and enteritis-related C. jejuni strains more than for the poultry strains [(31/38 (82 %) vs. 30/50 (60 %), p < 0.05]. Multilocus sequence typing (MLST) defined 15 sequence types (STs) and six clonal complexes (CCs) among poultry isolates, including one ST-3740 not previously documented. The most commonly identified type, ST-5 (13/66), in chicken was seen only once among human isolates (1/49) (p < 0.001). Amplified fragment length polymorphism (AFLP) revealed three major clusters (A, B, and C) among C. jejuni isolated from humans and poultry. There seems to be a lack of overlap between the major human and chicken clones, which suggests that there may be additional sources for campylobacteriosis other than poultry in Bangladesh. PMID:24962195

  11. 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.

  12. 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. PMID:25627752

  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. PMID:27231016

  14. Identification of Campylobacter jejuni on the basis of a species-specific gene that encodes a membrane protein.

    PubMed Central

    Stucki, U; Frey, J; Nicolet, J; Burnens, A P

    1995-01-01

    To facilitate discrimination between the closely related enteropathogens Campylobacter jejuni and C. coli, unique differences in antigenic surface structure were examined. A genomic library of C. jejuni 81116 was constructed in plasmid pBluescriptIISK- and expressed in Escherichia coli K-12. Rabbit hyperimmune serum raised against C. jejuni ATCC 29428 recognized a clone expressing a C. jejuni 24-kDa membrane-associated protein. Antiserum raised against sonicated recombinant E. coli expressing the 24-kDa protein reacted with C. jejuni, whereas C. coli did not react specifically. Determination of the nucleotide sequence of the DNA insert of this recombinant plasmid revealed an open reading frame encoding 214 amino acids; the gene was designated mapA; and its gene product was designated MAPA. The 18 N-terminal amino acid residues constitute a signal sequence characteristic of prokaryotic membrane lipoproteins. In a dot blot hybridization assay with a mapA probe, 120 clinical isolates of C. jejuni were unequivocally discriminated from 126 other campylobacters, including 34 C. coli isolates. A PCR test based on the mapA sequence was developed for identification of C. jejuni. A PCR product was obtained with all of the clinical isolates of C. jejuni tested from human, dog, cat, bovine calf, and chicken sources. Recombinant MAPA with an added C-terminal six-histidine tail was affinity purified and used to immunize rabbits. The rabbit anti-MAPA serum specifically recognized the protein in whole cells of C. jejuni on Western blots (immunoblots). The MAPA protein was present in all of the C. jejuni strains tested and was absent in C. coli and related campylobacters. PMID:7790451

  15. Antibiotic Resistance of Campylobacter jejuni and C. coli Isolated from Children with Diarrhea in Thailand and Japan.

    PubMed

    Pham, Ngan Thi Kim; Thongprachum, Aksara; Tran, Dinh Nguyen; Nishimura, Shuichi; Shimizu-Onda, Yuko; Trinh, Quang Duy; Khamrin, Pattara; Ukarapol, Nuthapong; Kongsricharoern, Tipachan; Komine-Aizawa, Shihoko; Okitsu, Shoko; Maneekarn, Niwat; Hayakawa, Satoshi; Ushijima, Hiroshi

    2016-01-01

    A total of 29 Campylobacter jejuni and C. coli strains were isolated from Thai and Japanese children with diarrhea using the Loop-mediated Isothermal Amplification method. The samples were evaluated for mutations in gyrA and 23S rRNA in order to assess resistance against fluoroquinolones and macrolides, respectively. Among the isolated strains, 9 (8 C. jejuni and 1 C. coli) were from Thai children, and the other 20 (C. jejuni) were isolated from Japanese children. High fluoroquinolone resistance rates were observed in Thai (66.7%) and Japanese (90%) children. Macrolide resistance was not observed in Japanese children but was observed at a considerable rate of 12.5% of C. jejuni isolated in the Thai cohort. The results indicate that continuous monitoring of resistance of Campylobacter strains to fluoroquinolones and macrolides is definitely necessary.

  16. Plant derived compounds inactivate antibiotic resistant Campylobacter jejuni strains

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Sixty-three Campylobacter isolates were screened for their resistance to the antibiotics ampicillin, cefaclor, ciprofloxacin, erythromycin, gentamycin, tetracycline, and trimethroprim/sulfamethoxazole. Based on this screen, the resistant strains D28a and H2a and the nonresistant strain A24a were se...

  17. Exchange of Lipooligosaccharide Synthesis Genes Creates Potential Guillain-Barré Syndrome-Inducible Strains of Campylobacter jejuni

    PubMed Central

    Phongsisay, Vongsavanh; Perera, Viraj N.; Fry, Benjamin N.

    2006-01-01

    Human ganglioside-like structures, such as GM1, found on some Campylobacter jejuni strains have been linked to inducing the Guillain-Barré Syndrome (GBS). This study shows that a C. jejuni strain without GM1-like molecules acquired large DNA fragments, including lipooligosaccharide synthesis genes, from a strain expressing GM1-like molecules and consequently transformed into a number of potential GBS-inducible transformants, which exhibited a high degree of genetic and phenotypic diversity. PMID:16428786

  18. High-resolution DNA melt curve analysis of the clustered, regularly interspaced short-palindromic-repeat locus of Campylobacter jejuni.

    PubMed

    Price, Erin P; Smith, Helen; Huygens, Flavia; Giffard, Philip M

    2007-05-01

    A novel method for genotyping the clustered, regularly interspaced short-palindromic-repeat (CRISPR) locus of Campylobacter jejuni is described. Following real-time PCR, CRISPR products were subjected to high-resolution melt (HRM) analysis, a new technology that allows precise melt profile determination of amplicons. This investigation shows that the CRISPR HRM assay provides a powerful addition to existing C. jejuni genotyping methods and emphasizes the potential of HRM for genotyping short sequence repeats in other species.

  19. A receptor-binding protein of Campylobacter jejuni bacteriophage NCTC 12673 recognizes flagellin glycosylated with acetamidino-modified pseudaminic acid.

    PubMed

    Javed, Muhammad Afzal; van Alphen, Lieke B; Sacher, Jessica; Ding, Wen; Kelly, John; Nargang, Cheryl; Smith, David F; Cummings, Richard D; Szymanski, Christine M

    2015-01-01

    Bacteriophage receptor-binding proteins (RBPs) confer host specificity. We previously identified a putative RBP (Gp047) from the campylobacter lytic phage NCTC 12673 and demonstrated that Gp047 has a broader host range than its parent phage. While NCTC 12673 recognizes the capsular polysaccharide (CPS) of a limited number of Campylobacter jejuni isolates, Gp047 binds to a majority of C. jejuni and related Campylobacter coli strains. In this study, we demonstrate that Gp047 also binds to acapsular mutants, suggesting that unlike the parent phage, CPS is not the receptor for Gp047. Affinity chromatography and far-western analyses of C. jejuni lysates using Gp047 followed by mass spectrometry indicated that Gp047 binds to the major flagellin protein, FlaA. Because C. jejuni flagellin is extensively glycosylated, we investigated this binding specificity further and demonstrate that Gp047 only recognizes flagellin decorated with acetamidino-modified pseudaminic acid. This binding activity is localized to the C-terminal quarter of the protein and both wild-type and coccoid forms of C. jejuni are recognized. In addition, Gp047 treatment agglutinates vegetative cells and reduces their motility. Because Gp047 is highly conserved among all campylobacter phages sequenced to date, it is likely that this protein plays an important role in the phage life cycle.

  20. A receptor-binding protein of Campylobacter jejuni bacteriophage NCTC 12673 recognizes flagellin glycosylated with acetamidino-modified pseudaminic acid.

    PubMed

    Javed, Muhammad Afzal; van Alphen, Lieke B; Sacher, Jessica; Ding, Wen; Kelly, John; Nargang, Cheryl; Smith, David F; Cummings, Richard D; Szymanski, Christine M

    2015-01-01

    Bacteriophage receptor-binding proteins (RBPs) confer host specificity. We previously identified a putative RBP (Gp047) from the campylobacter lytic phage NCTC 12673 and demonstrated that Gp047 has a broader host range than its parent phage. While NCTC 12673 recognizes the capsular polysaccharide (CPS) of a limited number of Campylobacter jejuni isolates, Gp047 binds to a majority of C. jejuni and related Campylobacter coli strains. In this study, we demonstrate that Gp047 also binds to acapsular mutants, suggesting that unlike the parent phage, CPS is not the receptor for Gp047. Affinity chromatography and far-western analyses of C. jejuni lysates using Gp047 followed by mass spectrometry indicated that Gp047 binds to the major flagellin protein, FlaA. Because C. jejuni flagellin is extensively glycosylated, we investigated this binding specificity further and demonstrate that Gp047 only recognizes flagellin decorated with acetamidino-modified pseudaminic acid. This binding activity is localized to the C-terminal quarter of the protein and both wild-type and coccoid forms of C. jejuni are recognized. In addition, Gp047 treatment agglutinates vegetative cells and reduces their motility. Because Gp047 is highly conserved among all campylobacter phages sequenced to date, it is likely that this protein plays an important role in the phage life cycle. PMID:25354466

  1. Campylobacter jejuni capsular genotypes are related to Guillain-Barré syndrome.

    PubMed

    Heikema, A P; Islam, Z; Horst-Kreft, D; Huizinga, R; Jacobs, B C; Wagenaar, J A; Poly, F; Guerry, P; van Belkum, A; Parker, C T; Endtz, H P

    2015-09-01

    In about one in a thousand cases, a Campylobacter jejuni infection results in the severe polyneuropathy Guillain-Barré syndrome (GBS). It is established that sialylated lipo-oligosaccharides (LOS) of C. jejuni are a crucial virulence factor in GBS development. Frequent detection of C. jejuni with sialylated LOS in stools derived from patients with uncomplicated enteritis implies that additional bacterial factors should be involved. To assess whether the polysaccharide capsule is a marker for GBS, the capsular genotypes of two geographically distinct GBS-associated C. jejuni strain collections and an uncomplicated enteritis control collection were determined. Capsular genotyping of C. jejuni strains from the Netherlands revealed that three capsular genotypes, HS1/44c, HS2 and HS4c, were dominant in GBS-associated strains and capsular types HS1/44c and HS4c were significantly associated with GBS (p 0.05 and p 0.01, respectively) when compared with uncomplicated enteritis. In a GBS-associated strain collection from Bangladesh, capsular types HS23/36c, HS19 and HS41 were most prevalent and the capsular types HS19 and HS41 were associated with GBS (p 0.008 and p 0.02, respectively). Next, specific combinations of the LOS class and capsular genotypes were identified that were related to the occurrence of GBS. Multilocus sequence typing revealed restricted genetic diversity for strain populations with the capsular types HS2, HS19 and HS41. We conclude that capsular types HS1/44c, HS2, HS4c, HS19, HS23/36c and HS41 are markers for GBS. Besides a crucial role for sialylated LOS of C. jejuni in GBS pathogenesis, the identified capsules may contribute to GBS susceptibility. PMID:26070960

  2. Phenotypic Characters and Molecular Epidemiology of Campylobacter Jejuni in East China.

    PubMed

    Zeng, Dexin; Zhang, Xiaoping; Xue, Feng; Wang, Yanhong; Jiang, Luyan; Jiang, Yuan

    2016-01-01

    In this study, we investigated the distribution, phenotypic and molecular typing characters of Campylobacter jejuni in domestic fowl, and livestock populations in East China, to provide some reference for researches on its molecular epidemiology. A total of 1250 samples were collected from different animal sources, and C. jejuni strains were then isolated and tested for antibiotic sensitivity. Antibiotics-resistance gene and pathogenic genes were detected by polymerase chain reaction. Phylogenic analysis on the C. jejuni strains was performed by multilocus sequence typing (MLST) method. The results showed that 108 out of the 1250 samples (mean 8.64%) were C. jejuni positive. These 108 C. jejuni strains were highly sensitive to antibiotics such as chloramphenicol, amoxicillin, amikacin, cefotaxime, and azithromycin, whereas they were highly resistant to antibiotics such as cefoperazone, cotrimoxazole, cefamandole, sulfamethoxazole, and cefradine. Pathogenicity related gene identification indicated that the mean carrying rate of adhesion related gene cadF and racR, flagellin gene flaA, toxin regulating gene cdtA, cdtB, cdtC, wlaN and virB11, heat shock proteins and transferring proteins related genes dnaJ and ceuE, CiaB and pldA were 92.45%, 38.69%, 73.58%, 71.70%, 52.83%, 96.23%, 12.26%, 1.89%, 0.94%, 65.09%, 39.62% and 9.43%, respectively. A total of 58.82% of these strains contained more than 6 pathogenicity-related genes. MLST typed 58 ST types from the 108 isolated C. jejuni strains, including 24 new types, and ST-21 was the major type, accounting for 39.3% of the total strains.

  3. Campylobacter jejuni capsular genotypes are related to Guillain-Barré syndrome.

    PubMed

    Heikema, A P; Islam, Z; Horst-Kreft, D; Huizinga, R; Jacobs, B C; Wagenaar, J A; Poly, F; Guerry, P; van Belkum, A; Parker, C T; Endtz, H P

    2015-09-01

    In about one in a thousand cases, a Campylobacter jejuni infection results in the severe polyneuropathy Guillain-Barré syndrome (GBS). It is established that sialylated lipo-oligosaccharides (LOS) of C. jejuni are a crucial virulence factor in GBS development. Frequent detection of C. jejuni with sialylated LOS in stools derived from patients with uncomplicated enteritis implies that additional bacterial factors should be involved. To assess whether the polysaccharide capsule is a marker for GBS, the capsular genotypes of two geographically distinct GBS-associated C. jejuni strain collections and an uncomplicated enteritis control collection were determined. Capsular genotyping of C. jejuni strains from the Netherlands revealed that three capsular genotypes, HS1/44c, HS2 and HS4c, were dominant in GBS-associated strains and capsular types HS1/44c and HS4c were significantly associated with GBS (p 0.05 and p 0.01, respectively) when compared with uncomplicated enteritis. In a GBS-associated strain collection from Bangladesh, capsular types HS23/36c, HS19 and HS41 were most prevalent and the capsular types HS19 and HS41 were associated with GBS (p 0.008 and p 0.02, respectively). Next, specific combinations of the LOS class and capsular genotypes were identified that were related to the occurrence of GBS. Multilocus sequence typing revealed restricted genetic diversity for strain populations with the capsular types HS2, HS19 and HS41. We conclude that capsular types HS1/44c, HS2, HS4c, HS19, HS23/36c and HS41 are markers for GBS. Besides a crucial role for sialylated LOS of C. jejuni in GBS pathogenesis, the identified capsules may contribute to GBS susceptibility.

  4. 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.

  5. Packaging of Campylobacter jejuni into Multilamellar Bodies by the Ciliate Tetrahymena pyriformis.

    PubMed

    Trigui, Hana; Paquet, Valérie E; Charette, Steve J; Faucher, Sébastien P

    2016-05-01

    Campylobacter jejuniis the leading cause of bacterial gastroenteritis worldwide. Transmission to humans occurs through consumption of contaminated food or water. The conditions affecting the persistence of C. jejuniin the environment are poorly understood. Some protozoa package and excrete bacteria into multilamellar bodies (MLBs). Packaged bacteria are protected from deleterious conditions, which increases their survival. We hypothesized that C. jejuni could be packaged under aerobic conditions by the amoeba Acanthamoeba castellanii or the ciliate Tetrahymena pyriformis, both of which are able to package other pathogenic bacteria.A. castellanii did not produce MLBs containing C. jejuni In contrast, when incubated with T. pyriformis,C. jejuni was ingested, packaged in MLBs, and then expelled into the milieu. The viability of the bacteria inside MLBs was confirmed by microscopic analyses. The kinetics of C. jejuni culturability showed that packaging increased the survival of C. jejuniup to 60 h, in contrast to the strong survival defect seen in ciliate-free culture. This study suggests that T. pyriformis may increase the risk of persistence of C. jejuniin the environment and its possible transmission between different reservoirs in food and potable water through packaging. PMID:26921427

  6. Comparison of epidemiologically linked Campylobacter jejuni isolated from human and poultry sources.

    PubMed

    Lajhar, S A; Jennison, A V; Patel, B; Duffy, L L

    2015-12-01

    Campylobacter jejuni is responsible for most foodborne bacterial infections worldwide including Australia. The aim of this study was to investigate a combination of typing methods in the characterization of C. jejuni isolated from clinical diarrhoeal samples (n = 20) and chicken meat (n = 26) in order to identify the source of infection and rank isolates based on their relative risk to humans. Sequencing of the flaA short variable region demonstrated that 86% of clinical isolates had genotypes that were also found in chicken meat. A polymerase chain reaction binary typing system identified 27 different codes based on the presence or absence of genes that have been reported to be associated with various aspects of C. jejuni pathogenicity, indicating that not all isolates may be of equal risk to human health. The lipooligosaccharide (LOS) of the C. jejuni isolates was classified into six classes (A, B, C, E, F, H) with 10·4% remaining unclassified. The majority (72·7%) of clinical isolates possessed sialylated LOS classes. Sialylated LOS classes were also detected in chicken isolates (80·7%). Antimicrobial tests indicated a low level of resistance, with no phenotypic resistance found to most antibiotics tested. A combination of typing approaches was useful to assign isolates to a source of infection and assess their risk to humans.

  7. Development of an immunoassay for rapid detection of ganglioside GM(1) mimicry in Campylobacter jejuni strains.

    PubMed

    Prendergast, M M; Kosunen, T U; Moran, A P

    2001-04-01

    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 GM(1) 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 (GM(1) ligand), peanut agglutinin (Gal beta1-->3GalNAc ligand), and anti-GM(1) 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-GM(1) ganglioside reagents. Subsequently, LPS extracts from 5 of 7 (71%) C. jejuni isolates and 2 of 3 (67%) C. jejuni culture collection strains bore GM(1) structures. Overall, the assay system was reliable, efficient, and reproducible and may be adapted for large-scale epidemiological studies.

  8. Participation of CheR and CheB in the chemosensory response of Campylobacter jejuni.

    PubMed

    Kanungpean, Doungjit; Kakuda, Tsutomu; Takai, Shinji

    2011-05-01

    Campylobacter jejuni is a leading cause of bacterial gastroenteritis in humans and a commensal bacterium of the intestinal tracts of animals, especially poultry. Chemotaxis is an important determinant for chicken colonization of C. jejuni. Adaptation has a crucial role in the gradient-sensing mechanism that underlies chemotaxis. The genome sequence of C. jejuni reveals the presence of genes encoding putative adaptation proteins, CheB and CheR. In-frame deletions of cheB, cheR and cheBR were constructed and the chemosensory behaviour of the resultant mutants was examined on swarm plates. CheB and CheR proteins significantly influence chemotaxis but are not essential for this behaviour to occur. Increased mobility of two methyl-accepting chemotaxis proteins (MCPs), DocC and Tlp1, during SDS-PAGE was detected in the mutants lacking functional CheB in the presence of CheR, presumably resulting from stable methylation of receptors. In vitro studies using tissue culture revealed that deletion of cheR resulted in hyperadherent and hyperinvasive phenotypes, while deletion of cheB resulted in nonadherent, noninvasive phenotypes. Furthermore, the ΔcheBR mutant showed significantly reduced ability to colonize chick caeca. Our data suggest that modification of chemoreceptors by the CheBR system is involved in regulation of chemotaxis in C. jejuni although CheB is apparently not controlled by phosphorylation. PMID:21292743

  9. The Role of Gelatinases in Campylobacter Jejuni Infection of Gnotobiotic Mice

    PubMed Central

    Alutis, Marie E.; Grundmann, Ursula; Fischer, André; Hagen, Ulrike; Kühl, Anja A.; Göbel, Ulf B.; Bereswill, Stefan; Heimesaat, Markus M.

    2015-01-01

    Matrix metalloproteinases (MMP)-2 and -9 (also referred to gelatinases-A and -B, respectively) are upregulated in the inflamed gut of mice and men. We recently demonstrated that synthetic gelatinase blockage reduced large intestinal pro-inflammatory immune responses and apoptosis following murine Campylobacter (C.) jejuni infection. In order to address which gelatinase mediates C. jejuni-induced immune responses, gnotobiotic MMP-2–/–, MMP-9–/–, and wildtype (WT) mice were generated by broadspectrum antibiotic treatment and perorally infected with C. jejuni strain 81-176. The pathogen stably colonized the murine intestinal tract irrespective of the genotype but did not translocate to extra-intestinal compartments. At days 8 and 14 postinfection (p.i.), less pronounced colonic histopathological changes were observed in infected MMP-2–/– mice, less distinct epithelial apoptosis, but more epithelial proliferation in both MMP-2–/– and MMP-9–/– mice, as compared to WT controls. Reduced immune responses in gelatinase-deficient mice were characterized by lower numbers of effector as well as innate and adaptive immune cells within the colonic mucosa and lamina propria. The expression of IL-22, IL-18, IL-17A, and IL-1β mRNA was higher in the colon of MMP-2–/– as compared to WT mice. In conclusion, both MMP-2 and MMP-9 are differentially involved in mediating C. jejuni-induced intestinal immunopathology. PMID:26716014

  10. Campylobacter jejuni Infections Associated with Raw Milk Consumption--Utah, 2014.

    PubMed

    Davis, Kenneth R; Dunn, Angela C; Burnett, Cindy; McCullough, Laine; Dimond, Melissa; Wagner, Jenni; Smith, Lori; Carter, Amy; Willardson, Sarah; Nakashima, Allyn K

    2016-04-01

    In May 2014, the Utah Public Health Laboratory (UPHL) notified the Utah Department of Health (UDOH) of specimens from three patients infected with Campylobacter jejuni yielding indistinguishable pulsed-field gel electrophoresis (PFGE) patterns. All three patients had consumed raw (unpasteurized and nonhomogenized) milk from dairy A. In Utah, raw milk sales are legal from farm to consumer with a sales permit from the Utah Department of Agriculture and Food (UDAF). Raw milk dairies are required to submit monthly milk samples to UDAF for somatic cell and coliform counts, both of which are indicators of raw milk contamination. Before this cluster's identification, dairy A's routine test results were within acceptable levels (<400,000 somatic cells/mL and <10 coliform colony forming units/mL). Subsequent enhanced testing procedures recovered C. jejuni, a fastidious organism, in dairy A raw milk; the isolate matched the cluster pattern. UDAF suspended dairy A's raw milk permit during August 4-October 1, and reinstated the permit when follow-up cultures were negative. Additional cases of C. jejuni infection were identified in October, and UDAF permanently revoked dairy A's permit to sell raw milk on December 1. During May 9-November 6, 2014, a total of 99 cases of C. jejuni infection were identified. Routine somatic cell and coliform counts of raw milk do not ensure its safety. Consumers should be educated that raw milk might be unsafe even if it meets routine testing standards.

  11. 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.

  12. Bile-induced 'pili' in Campylobacter jejuni are bacteria-independent artifacts of the culture medium.

    PubMed

    Gaynor, E C; Ghori, N; Falkow, S

    2001-03-01

    In 1996, it was reported that the enteric pathogen Campylobacter jejuni produces pilus-like appendages in response to bile salts such as deoxycholate (DOC), and that the formation of these appendages requires the putative peptidase PspA. Pili were known to be important virulence determinants in other pathogenic bacteria but had never before been observed for C. jejuni. We report here that these appendages are not pili, but are instead a bacteria-independent morphological artifact of the growth medium. Furthermore, the pspA gene is not required for their formation. Broth cultures containing a threshold concentration of DOC inoculated with no bacteria produced identical abundant, fibrous, pilus-like structures as those cultures that had been inoculated with C. jejuni. These fibres were also found in growth media from DOC-containing pspA:CmR mutant cultures. These results are consistent with the absence of candidate pilin monomers in protein gel analyses as well as the dearth of pilin-like genes and pilus formation gene clusters in the C. jejuni genome.

  13. Epidemiology of Campylobacter jejuni Outbreak in a Middle School in Incheon, Korea

    PubMed Central

    Kim, Na-Yeon; Cho, Nam-Gue; Kim, Jung-Hee; Kang, Young-Ah; Lee, Ha-Gyung

    2010-01-01

    On July 6, 2009, an outbreak of gastroenteritis occurred among middle school students in Incheon. An investigation to identify the source and describe the extent of the outbreak was conducted. A retrospective cohort study among students, teachers, and food handlers exposed to canteen food in the middle school was performed. Using self-administered questionnaires, information was collected concerning on symptoms, days that canteen food was consumed, and food items consumed. Stool samples were collected from 66 patients and 11 food handlers. The catering kitchen was inspected and food samples were taken. Of the 791 people who ate canteen food, 92 cases became ill, representing an attack rate of 11.6%. Thirty-one (40.3%) of the 77 stool specimens were positive for Campylobacter jejuni. Interviews with kitchen staff indicated the likelihood that undercooked chicken was provided. This is the first recognized major C. jejuni outbreak associated with contaminated chicken documented in Korea. PMID:21060748

  14. Campylobacter jejuni bacteremia and Guillain-Barré syndrome in a renal transplant recipient.

    PubMed

    Maccario, M; Tarantino, A; Nobile-Orazio, E; Ponticelli, C

    1998-01-01

    In patients who have not undergone transplantation, Guillain-Barré syndrome (GBS) is typically preceded by an acute infection often sustained by Campylobacter jejuni. Thus far, in renal transplant recipients, only eight cases of GBS have been reported. In seven patients GBS was attributed to cytomegalovirus infection and in the eighth patient to cyclosporin A neurotoxicity. We report here the case of a GBS in a renal transplant recipient following C. jejuni bacteremia. The infection quickly disappeared after erythromycin and methronidazole therapy. GBS progressively evolved into a paraparesis within 1 week. After reaching a plateau phase, the clinical status improved and the patient was able to walk unassisted after 3 weeks. At this last check-up, 54 months later, the patient was doing well with a functioning graft and only minimal weakness of the lower limbs.

  15. Development, stability, and molecular mechanisms of macrolide resistance in Campylobacter jejuni.

    PubMed

    Caldwell, Dave Bryson; Wang, Ying; Lin, Jun

    2008-11-01

    Previous studies of macrolide resistance in Campylobacter were primarily focused on strains from various origins or used in vitro systems. In this study, we conducted both in vitro and in vivo experiments to examine the development, stability, and genetic basis of macrolide resistance in Campylobacter jejuni using erythromycin-resistant (Ery(r)) mutants derived from the same parent strain. Chickens inoculated with low-level Ery(r) mutants (MIC = 32 or 64 microg/ml) at 15 days old did not shed highly Ery(r) mutants (MIC > 512 microg/ml) after prolonged exposure to a low dose of tylosin. The low-level Ery resistance was not stable in vitro or in vivo in the absence of macrolide selection pressure. However, high-level Ery resistance displayed remarkable stability in vitro and in vivo. Ribosomal sequence analysis of 69 selected Ery(r) mutants showed that specific point mutations (A2074G or A2074C) occurred in all highly Ery(r) mutants. No mutations in ribosomal protein L4 were observed in any of the in vitro-selected Ery(r) mutants. However, three specific mutations in L4, G74D, G57D, and G57V, were widely found among in vivo-selected Ery(r) mutants. Insertion of three amino acids, TSH, at position 98 in ribosomal protein L22 was observed only in mutants selected in vitro. Inactivation of the CmeABC efflux pump dramatically reduced Ery MICs in Ery(r) mutants. Together, these findings suggest that multiple factors contribute to the emergence of highly Ery(r) Campylobacter in chicken, reveal resistance level-dependent stability of macrolide resistance in C. jejuni, and indicate that C. jejuni utilizes complex and different mechanisms to develop Ery resistance in vitro and in vivo. PMID:18779354

  16. Avian wildlife reservoir of Campylobacter fetus subsp. jejuni, Yersinia spp., and Salmonella spp. in Norway.

    PubMed

    Kapperud, G; Rosef, O

    1983-02-01

    Cloacal swabs from 540 wild-living birds were cultured for Campylobacter fetus subsp. jejuni, Yersinia spp., and Salmonella spp. The carrier rates detected were as follows: C. fetus subsp. jejuni, 28.4%; Yersinia spp., 1.2%; and Salmonella spp., 0.8%. All birds were apparently healthy when captured. C. fetus subsp. jejuni was isolated from 11 of the 40 bird species examined. Among birds inhabiting the city of Oslo, the highest isolation rate was found in crows (Corvus corone cornix) (89.8%), followed by gulls (Larus spp.) (50.0%) and domestic pigeons (Columba livia domesticus) (4.2%). The gulls and crows scavenge on refuse dumps. High carrier rates were also detected among the following birds from nonurban, coastal areas: puffin (Fratercula arctica) (51.3%), common tern (Sterna hirundo) (5.6%), common gull (Larus canus) (18.9%), black-headed gull (Larus ridibundus) (13.2%), and herring gull (Larus argentatus) (4.2%). The list of species harboring C. fetus subsp. jejuni also includes the Ural owl (Strix uralensis), goldeneye (Bucephala clangula), and reed bunting (Emberiza schoeniclus). The following five Yersinia strains were isolated: Y. kristensenii (two strains), Y. intermedia (two strains), and "Yersinia X2" (one strain). Four strains belonging to the genus Salmonella were isolated from three different species of gulls. These isolates were identified as S. typhimurium, S. indiana, and S. djugu. The results indicate that campylobacters are a normal component of the intestinal flora in several bird species, whereas Salmonella and Yersinia carriers are more sporadic. PMID:6338824

  17. Characterization of the oxidative stress stimulon and PerR regulon of Campylobacter jejuni

    PubMed Central

    Palyada, Kiran; Sun, Yi-Qian; Flint, Annika; Butcher, James; Naikare, Hemant; Stintzi, Alain

    2009-01-01

    Background During gut colonization, the enteric pathogen Campylobacter jejuni must surmount the toxic effects of reactive oxygen species produced by its own metabolism, the host immune system, and intestinal microflora. Elucidation of C. jejuni oxidative stress defense mechanisms is critical for understanding Campylobacter pathophysiology. Results The mechanisms of oxidative stress defense in C. jejuni were characterized by transcriptional profiling and phenotypic analysis of wild-type and mutant strains. To define the regulon of the peroxide-sensing regulator, PerR, we constructed an isogenic ΔperR mutant and compared its transcriptome profile with that of the wild-type strain. Transcriptome profiling identified 104 genes that belonged to the PerR regulon. PerR appears to regulate gene expression in a manner that both depends on and is independent of the presence of iron and/or H2O2. Mutation of perR significantly reduced motility. A phenotypic analysis using the chick colonization model showed that the ΔperR mutant exhibited attenuated colonization behavior. An analysis of changes in the transcriptome induced by exposure to H2O2, cumene hydroperoxide, or menadione revealed differential expression of genes belonging to a variety of biological pathways, including classical oxidative stress defense systems, heat shock response, DNA repair and metabolism, fatty acid biosynthesis, and multidrug efflux pumps. Mutagenic and phenotypic studies of the superoxide dismutase SodB, the alkyl-hydroxyperoxidase AhpC, and the catalase KatA, revealed a role for these proteins in oxidative stress defense and chick gut colonization. Conclusion This study reveals an interplay between PerR, Fur, iron metabolism and oxidative stress defense, and highlights the role of these elements in C. jejuni colonization of the chick cecum and/or subsequent survival. PMID:19835633

  18. A PAS domain-containing regulator controls flagella-flagella interactions in Campylobacter jejuni

    PubMed Central

    Reuter, Mark; Periago, Paula M.; Mulholland, Francis; Brown, Helen L.; van Vliet, Arnoud H. M.

    2015-01-01

    The bipolar flagella of the foodborne bacterial pathogen Campylobacter jejuni confer motility, which is essential for virulence. The flagella of C. jejuni are post-translationally modified, but how this process is controlled is not well understood. In this work, we have identified a novel PAS-domain containing regulatory system, which modulates flagella-flagella interactions in C. jejuni. Inactivation of the cj1387c gene, encoding a YheO-like PAS6 domain linked to a helix-turn-helix domain, resulted in the generation of a tightly associated “cell-train” morphotype, where up to four cells were connected by their flagella. The morphotype was fully motile, resistant to vortexing, accompanied by increased autoagglutination, and was not observed in aflagellated cells. The Δcj1387c mutant displayed increased expression of the adjacent Cj1388 protein, which comprises of a single endoribonuclease L-PSP domain. Comparative genomics showed that cj1387c (yheO) orthologs in bacterial genomes are commonly linked to an adjacent cj1388 ortholog, with some bacteria, including C. jejuni, containing another cj1388-like gene (cj0327). Inactivation of the cj1388 and cj0327 genes resulted in decreased autoagglutination in Tween-20-supplemented media. The Δcj1388 and Δcj0327 mutants were also attenuated in a Galleria larvae-based infection model. Finally, substituting the sole cysteine in Cj1388 for serine prevented Cj1388 dimerization in non-reducing conditions, and resulted in decreased autoagglutination in the presence of Tween-20. We hypothesize that Cj1388 and Cj0327 modulate post-translational modification of the flagella through yet unidentified mechanisms, and propose naming Cj1387 the Campylobacter Flagella Interaction Regulator CfiR, and the Cj1388 and Cj0327 protein as CfiP and CfiQ, respectively. PMID:26284050

  19. Hygromycin B and Apramycin Antibiotic Resistance Cassettes for Use in Campylobacter jejuni

    PubMed Central

    Cameron, Andrew; Gaynor, Erin C.

    2014-01-01

    Campylobacter jejuni genetic manipulation is restricted by the limited number of antibiotic resistance cassettes available for use in this diarrheal pathogen. In this study, two antibiotic resistance cassettes were developed, encoding for hygromycin B and apramycin resistance, for use in mutagenesis or for selection of gene expression and complementation constructs in C. jejuni. First, the marker genes were successfully modified to allow for insertional mutagenesis or deletion of a gene-of-interest, and were bracketed with restriction sites for the facilitation of site-specific cloning. These hygromycin B and apramycin markers are encoded by plasmids pAC1H and pAC1A, respectively. We also modified an insertional gene-delivery vector to create pRRH and pRRA, containing the hygromycin B and apramycin resistance genes, and 3 unique restriction sites for the directional introduction of genes into the conserved multi-copy rRNA gene clusters of the C. jejuni chromosome. We determined the effective antibiotic concentrations required for selection, and established that no harmful effects or fitness costs were associated with carrying hygromycin B or apramycin resistance under standard C. jejuni laboratory conditions. Using these markers, the arylsulfatase reporter gene astA was deleted, and the ability to genetically complement the astA deletion using pRRH and pRRA for astA gene insertion was demonstrated. Furthermore, the relative levels of expression from the endogenous astA promoter were compared to that of polycistronic mRNA expression from the constitutive promoter upstream of the resistance gene. The development of additional antibiotic resistance cassettes for use in Campylobacter will enable multiple gene deletion and expression combinations as well as more in-depth study of multi-gene systems important for the survival and pathogenesis of this important bacterium. PMID:24751825

  20. Avian wildlife reservoir of Campylobacter fetus subsp. jejuni, Yersinia spp., and Salmonella spp. in Norway.

    PubMed

    Kapperud, G; Rosef, O

    1983-02-01

    Cloacal swabs from 540 wild-living birds were cultured for Campylobacter fetus subsp. jejuni, Yersinia spp., and Salmonella spp. The carrier rates detected were as follows: C. fetus subsp. jejuni, 28.4%; Yersinia spp., 1.2%; and Salmonella spp., 0.8%. All birds were apparently healthy when captured. C. fetus subsp. jejuni was isolated from 11 of the 40 bird species examined. Among birds inhabiting the city of Oslo, the highest isolation rate was found in crows (Corvus corone cornix) (89.8%), followed by gulls (Larus spp.) (50.0%) and domestic pigeons (Columba livia domesticus) (4.2%). The gulls and crows scavenge on refuse dumps. High carrier rates were also detected among the following birds from nonurban, coastal areas: puffin (Fratercula arctica) (51.3%), common tern (Sterna hirundo) (5.6%), common gull (Larus canus) (18.9%), black-headed gull (Larus ridibundus) (13.2%), and herring gull (Larus argentatus) (4.2%). The list of species harboring C. fetus subsp. jejuni also includes the Ural owl (Strix uralensis), goldeneye (Bucephala clangula), and reed bunting (Emberiza schoeniclus). The following five Yersinia strains were isolated: Y. kristensenii (two strains), Y. intermedia (two strains), and "Yersinia X2" (one strain). Four strains belonging to the genus Salmonella were isolated from three different species of gulls. These isolates were identified as S. typhimurium, S. indiana, and S. djugu. The results indicate that campylobacters are a normal component of the intestinal flora in several bird species, whereas Salmonella and Yersinia carriers are more sporadic.

  1. Molecular and Epidemiological Analysis of a Campylobacter jejuni Outbreak in Northern Italy in November 2013.

    PubMed

    Lucarelli, Claudia; Dionisi, Anna Maria; Trezzi, Livia; Farina, Claudio; Passera, Marco; Kärki, Tommi; D'Ancona, Fortunato; Luzzi, Ida

    2016-09-01

    Campylobacter spp. is the most common gastrointestinal pathogen worldwide with a very low reported incidence in Italy. In November of 2013, local and national public health authorities investigated an outbreak caused by Campylobacter jejuni among children in a kindergarten in Northern Italy. A case was defined as a child who had diarrhea with a laboratory-confirmed diagnosis of C. jejuni between 11 and 30 November. Stool samples from the kindergarten kitchen staff and environmental samples from the kitchen were examined for enteric pathogens. As food leftovers were not available, the menu logbook of the refectory was reviewed to identify a possible source of the outbreak. C. jejuni strains were tested for antimicrobial susceptibility and subtyped by pulsed field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). We identified 20 cases among 247 schoolchildren (attack rate = 8%), all who reported having lunch in the kindergarten. The stools from the kitchen staff as well as the environmental samples were negative for enteric pathogens. The identified outbreak strains (n = 5) were sensitive to all of the antimicrobials tested; the first four strains showed an identical PFGE profile, whereas the fifth strain showed a PFGE pattern similarity of 89%. Using MLST, all five strains were assigned to a single sequence type (ST), ST451 (clonal complex, CC21); this was the first identification of ST and the third reported outbreak of C. jejuni in Italy. Molecular typing confirmed that most of the cases belonged to a clonal cluster supporting the hypothesis of a common source; however, the source was not identified. Due to a delayed start of the investigation, it was not possible to perform any microbiological evaluation of the food consumed. PMID:27455195

  2. Molecular evidence for the thriving of Campylobacter jejuni ST-4526 in Japan.

    PubMed

    Asakura, Hiroshi; Brüggemann, Holger; Sheppard, Samuel K; Ekawa, Tomoya; Meyer, Thomas F; Yamamoto, Shigeki; Igimi, Shizunobu

    2012-01-01

    Campylobacter jejuni is a leading cause of human gastroenteritis worldwide. This study aimed at a better understanding of the genetic diversity of this pathogen disseminated in Japan. We performed multilocus sequence typing (MLST) of Campylobacter jejuni isolated from different sources (100 human, 61 poultry, and 51 cattle isolates) in Japan between 2005 and 2006. This approach identified 62 sequence types (STs) and 19 clonal complexes (CCs), including 11 novel STs. These 62 STs were phylogenetically divided into 6 clusters, partially exhibiting host association. We identified a novel ST (ST-4526) that has never been reported in other countries; a phylogenetic analysis showed that ST-4526 and related STs showed distant lineage from the founder ST, ST-21 within CC-21. Comparative genome analysis was performed to investigate which properties could be responsible for the successful dissemination of ST-4526 in Japan. Results revealed that three representative ST-4526 isolates contained a putative island comprising the region from Cj0737 to Cj0744, which differed between the ST-4526 isolates and the reference strain NCTC11168 (ST-43/CC-21). Amino acid sequence alignment analyses showed that two of three ST-4526 isolates expressed 693aa- filamentous hemagglutination domain protein (FHA), while most of other C. jejuni strains whose genome were sequenced exhibited its truncation. Correspondingly, host cell binding of FHA-positive C. jejuni was greater than that of FHA-truncated strains, and exogenous administration of rFHA protein reduced cell adhesion of FHA-positive bacteria. Biochemical assays showed that this putative protein exhibited a dose-dependent binding affinity to heparan sulfate, indicating its adhesin activity. Moreover, ST-4526 showed increased antibiotic-resistance (nalidixic acid and fluoroquinolones) and a reduced ability for DNA uptake. Taken together, our data suggested that these combined features contributed to the clonal thriving of ST-4526 in Japan.

  3. Effects of decontamination at varying contamination levels of Campylobacter jejuni on broiler meat.

    PubMed

    Boysen, L; Wechter, N S; Rosenquist, H

    2013-05-01

    When assessing effects of decontamination techniques on counts of Campylobacter spp. on broiler meat, it is essential that the results reflect the variations that may exist. Decontamination studies often use high inoculation levels (10(7) to 10(8) cfu) and one or few strains of Campylobacter jejuni, thereby restricting the results to reflect only a limited part of the true situation. This study presents results from physical and chemical decontamination of broiler meat medallions using different strains and different initial concentrations of C. jejuni. For 3 strains of C. jejuni, mean log reductions obtained by freezing at -20°C for 7 d was significantly higher for an initial concentration of 10(7) cfu/sample on the meat compared with an initial concentration of 10(3) cfu/sample. For freezing at -20°C for 24 h or application of 6% tartaric acid and subsequent storage for 24 h, no statistically significant difference in reductions was found for initial concentrations ranging from 10(3) to 10(7) cfu per sample. The mean log reductions obtained by all techniques were strongly dependent on the strain tested. The results reveal that reductions obtained with high inoculation levels of C. jejuni (10(7) cfu/sample) or single or few strains of the species (or both) should not be interpreted as a generic result for the species. If inoculation studies cannot be replaced by investigations of naturally contaminated meat, we advise using a mixture of strains found in the production environment at levels as close as possible to the natural contamination level.

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

    PubMed Central

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

    2016-01-01

    Summary 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. PMID:27043446

  5. Molecular identification of Campylobacter jejuni and coli from chicken, calves and dogs to determine its potential threat on human being

    PubMed Central

    Begum, Sonuwara; Sekar, M.; Gunaseelan, L.; Gawande, Monica; Suganya, G.; Malar, P. Annal Selva; Karthikeyan, A.

    2015-01-01

    Aim: Campylobacter is an emerging zoonotic pathogen and one of the leading cause of foodborne infection worldwide and it has been isolated from a variety of animal species. The aim of this study was to identify Campylobacter jejuni and Campylobacter coli from dogs, calves, and poultry using polymerase chain reaction (PCR). Methodology: A total of 104 number of samples comprising cloacal swab from poultry (38), a rectal swab from dogs (40), and calves (26) were collected for the isolation of thermophilic Campylobacters using conventional culture method. PCR was used for identification of mapA gene for C.jejuni and ceuE for C.coli. Results: The overall presence of Campylobacter was found to be 67(64.42%) from the samples, out of which 6 isolates belongs to C. jejuni species, were 5(18.51%) from chicken and 1(4.17%) from dog was recorded and about 17 isolates belongs to C. coli species were 9(33.33%), 6 (25%), and 1(9.09%) from chicken, dog and calves was recorded. Conclusion: Results suggested that Campylobacter reservoirs chicken, calves and pet dogs can play a role as the source of infection to human beings and PCR can be an ideal tool for molecular confirmation at the species level. PMID:27047055

  6. Campylobacter jejuni contamination of broiler carcasses: Population dynamics and genetic profiles at slaughterhouse level.

    PubMed

    Gruntar, Igor; Biasizzo, Majda; Kušar, Darja; Pate, Mateja; Ocepek, Matjaž

    2015-09-01

    Six slaughter batches deriving from six typical industrial broiler flocks were examined for the presence, quantity and genetic characteristics of contaminating Campylobacter jejuni (C. jejuni) during various stages of slaughtering and carcass processing. To assess the contamination dynamics of the carcasses, the analyses were always conducted on neck-skin samples from the same pre-selected and carefully marked carcasses in each batch. The skin samples were taken sequentially at three successive slaughter-line locations in the evisceration room, after three-day refrigeration and after three-day freezing procedure. Caecal samples from the same animals were also tested, as well as samples from the slaughterhouse environment before and after slaughtering. The samples were analysed by the ISO10272 isolation method; campylobacters from neck-skin samples were also quantified. Isolates were species-identified and genotyped by pulsed-field gel electrophoresis (PFGE). On average, the highest C. jejuni skin contamination was detected at the first sampling point (post-plucking), suggesting that the majority of Campylobacter contamination actually occurs before the entrance to the eviscerating room, probably during the preceding plucking stage. In two out of five positive batches, an additional increase in contamination was recorded after the evisceration step. An evident trend of increasing contamination level was detected when successive batches were compared at each of two initial sampling sites in the evisceration room, indicating an accumulation of contaminating C. jejuni at some point before the evisceration room. Three-day refrigeration and three-day freezing caused a 4.5- and 142-fold drop in mean C. jejuni CFU counts, respectively. All pre-slaughtering samples from the slaughterhouse environment were negative and all post-slaughtering samples, except water from the scalding tank, were positive. Pulsotypes were limited: altogether five different types were detected

  7. Campylobacter jejuni contamination of broiler carcasses: Population dynamics and genetic profiles at slaughterhouse level.

    PubMed

    Gruntar, Igor; Biasizzo, Majda; Kušar, Darja; Pate, Mateja; Ocepek, Matjaž

    2015-09-01

    Six slaughter batches deriving from six typical industrial broiler flocks were examined for the presence, quantity and genetic characteristics of contaminating Campylobacter jejuni (C. jejuni) during various stages of slaughtering and carcass processing. To assess the contamination dynamics of the carcasses, the analyses were always conducted on neck-skin samples from the same pre-selected and carefully marked carcasses in each batch. The skin samples were taken sequentially at three successive slaughter-line locations in the evisceration room, after three-day refrigeration and after three-day freezing procedure. Caecal samples from the same animals were also tested, as well as samples from the slaughterhouse environment before and after slaughtering. The samples were analysed by the ISO10272 isolation method; campylobacters from neck-skin samples were also quantified. Isolates were species-identified and genotyped by pulsed-field gel electrophoresis (PFGE). On average, the highest C. jejuni skin contamination was detected at the first sampling point (post-plucking), suggesting that the majority of Campylobacter contamination actually occurs before the entrance to the eviscerating room, probably during the preceding plucking stage. In two out of five positive batches, an additional increase in contamination was recorded after the evisceration step. An evident trend of increasing contamination level was detected when successive batches were compared at each of two initial sampling sites in the evisceration room, indicating an accumulation of contaminating C. jejuni at some point before the evisceration room. Three-day refrigeration and three-day freezing caused a 4.5- and 142-fold drop in mean C. jejuni CFU counts, respectively. All pre-slaughtering samples from the slaughterhouse environment were negative and all post-slaughtering samples, except water from the scalding tank, were positive. Pulsotypes were limited: altogether five different types were detected

  8. Rapid identification of Campylobacter jejuni from poultry carcasses and slaughtering environment samples by real-time PCR.

    PubMed

    Ivanova, Mirena; Singh, Randhir; Dharmasena, Muthu; Gong, Chao; Krastanov, Albert; Jiang, Xiuping

    2014-06-01

    The objective of this study was to develop a real-time PCR assay for rapid identification of Campylobacter jejuni and to apply the method in analyzing samples from poultry processing. A C. jejuni-specific primer set targeting a portion of the C. jejuni hippuricase gene was developed. The specificity of the newly designed primer pair was verified using 5 C. jejuni strains and 20 other bacterial strains. Sensitivity was determined to be as low as 1 genome copy per reaction. A total of 73 samples were collected at different sites along the processing line during 2 visits to a poultry slaughterhouse and were examined by direct plating onto modified charcoal cefoperazone deoxycholate agar or after enrichment in Bolton broth followed by plating on modified charcoal cefoperazone deoxycholate agar. The newly developed real-time PCR assay was used to identify the presumptive colonies as belonging to C. jejuni. A real-time PCR assay targeting 16S ribosomal RNA was also applied to determine Campylobacter spp. prevalence. Results from the real-time PCR analysis indicated considerable variability in Campylobacter contamination, with incidence rates of 72.7 and 27.6% for sampling days A and B, respectively. Campylobacter was isolated from 100% of prescalded and preeviscerated carcasses on sampling day A. In contrast, on sampling day B, the highest number of Campylobacter-positive carcasses was recovered after evisceration (60%). The chilling process significantly reduced (P < 0.05) Campylobacter population, but the percentage of positive samples on sampling day A increased to 80%. All samples collected from the processing environment, except scalding tank 3 and the prechiller and chiller tanks, were 100% positive on day A, whereas no campylobacters were isolated from machinery on sampling day B. Our results revealed the widespread of C. jejuni in poultry processing and proved that the newly developed real-time PCR assay is a simple, specific, and inexpensive method for rapid C

  9. Prevalence and genotypes of Campylobacter jejuni from urban environmental sources in comparison with clinical isolates from children.

    PubMed

    Ramonaite, Sigita; Kudirkiene, Egle; Tamuleviciene, Egle; Leviniene, Giedra; Malakauskas, Alvydas; Gölz, Greta; Alter, Thomas; Malakauskas, Mindaugas

    2014-09-01

    This study aimed to investigate the prevalence of Campylobacter jejuni in potential contamination sources that are not regularly monitored such as free-living urban pigeons and crows, dogs, cats and urban environmental water and to assess the possible impact on the epidemiology of campylobacteriosis in children using multilocus sequence typing (MLST). Campylobacter spp. were detected in 36.2 % of faecal samples of free-living urban birds and in 40.4 % of environmental water samples. A low prevalence of Campylobacter spp. was detected in dogs and cats, with 7.9 and 9.1 %, respectively. Further identification of isolates revealed that environmental water and pet samples were mostly contaminated by other Campylobacter spp. than C. jejuni, whereas C. jejuni was the most prevalent species in faecal samples of free-living birds (35.4 %). This species was the dominant cause of campylobacteriosis in children (91.5 %). In addition, the diversity of C. jejuni MLST types in free-living birds and children was investigated. Clonal complex (CC) 179 was predominant among free-living urban birds; however, only two isolates from children were assigned to this CC. One dog and one child isolate were assigned to the same clonal complex (CC48) and sequence type (ST) 918. The dominant two clonal complexes among the child clinical isolates (CC353 and CC21) were not detected among C. jejuni strains isolated from environmental sources examined in this study. As only two CCs were shared by environmental and child C. jejuni isolates and a high number of novel alleles and STs were found in C. jejuni isolated from free-living urban birds and environmental water, there is probably only a limited link between urban environmental sources and campylobacteriosis in children, particularly in rather cold climatic conditions.

  10. Development and evaluation of immunochromatographic assay for simple and rapid detection of Campylobacter jejuni and Campylobacter coli in human stool specimens.

    PubMed

    Kawatsu, Kentaro; Kumeda, Yuko; Taguchi, Masumi; Yamazaki-Matsune, Wataru; Kanki, Masashi; Inoue, Kiyoshi

    2008-04-01

    An immunochromatographic assay (Campy-ICA) using a newly generated single monoclonal antibody against a 15-kDa cell surface protein of Campylobacter jejuni was developed. When cell suspensions of 86 C. jejuni strains and 27 Campylobacter coli strains were treated with a commercially available bacterial protein extraction reagent and the resulting extracts were tested with the Campy-ICA, they all yielded positive results. The minimum detectable limits for the C. jejuni strains ranged from 1.8 x 10(4) to 8.2 x 10(5) CFU/ml of cell suspension, and those for the C. coli strains ranged from 1.4 x 10(5) to 4.6 x 10(6) CFU/ml of cell suspension. All 26 non-Campylobacter species tested yielded negative results with the Campy-ICA. To evaluate the ability of the Campy-ICA to detect C. jejuni and C. coli in human stool specimens, suspensions of 222 stool specimens from patients with acute gastroenteritis were treated with the bacterial protein extraction reagent, and the resulting extracts were tested with the Campy-ICA. The Campy-ICA results showed a sensitivity of 84.8% (28 of 33 specimens) and a specificity of 100% (189 of 189 specimens) compared to the results of isolation of C. jejuni and C. coli from the stool specimens by a bacterial culture test. The Campy-ICA was simple to perform and was able to detect Campylobacter antigen in a fecal extract within 15 min. These results suggest that Campy-ICA testing of fecal extracts may be useful as a simple and rapid adjunct to stool culture for detecting C. jejuni and C. coli in human stool specimens.

  11. 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.

  12. Influence of bird strain on competitive exclusion of Campylobacter jejuni in young chicks.

    PubMed

    Laisney, M J; Gillard, M O; Salvat, G

    2004-02-01

    1. Newly hatched chicks of either layer or broiler strain were treated orally at regular intervals with either homologous or heterologous gut-flora preparations from young donor birds, in an attempt to prevent subsequent colonisation with Campylobacter jejuni by 'competitive exclusion' (CE). 2. Donors of 3 to 10 d of age were chosen to correspond with the period in which intensively reared poultry are least likely to become colonised with Campylobacter. 3. In two separate trials, material from donor layer hens (ISA Brown) protected male chicks of the same strain against a low (195 to 360 cfu/bird) Campylobacter challenge, but the same kind of material was ineffective when administered to chicks of a broiler strain (JA957). 4. Two further trials involved treatment preparations from young broilers, which failed to prevent Campylobacter colonisation of broiler chicks, even when colonisation occurred relatively slowly from a challenge of 90 to 94 cfu/bird. 5. It was concluded that any CE effect observed was strongly dependent on bird strain.

  13. 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

  14. 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. PMID:26161743

  15. Cj1136 Is Required for Lipooligosaccharide Biosynthesis, Hyperinvasion, and Chick Colonization by Campylobacter jejuni

    PubMed Central

    Javed, Muhammad Afzal; Cawthraw, Shaun A.; Baig, Abiyad; Li, Jianjun; McNally, Alan; Oldfield, Neil J.; Newell, Diane G.

    2012-01-01

    Campylobacter jejuni is a major cause of bacterial food-borne enteritis worldwide, and invasion into intestinal epithelial cells is an important virulence mechanism. Recently we reported the identification of hyperinvasive C. jejuni strains and created a number of transposon mutants of one of these strains, some of which exhibited reduced invasion into INT-407 and Caco-2 cells. In one such mutant the transposon had inserted into a homologue of cj1136, which encodes a putative galactosyltransferase according to the annotation of the C. jejuni NCTC11168 genome. In the current study, we investigated the role of cj1136 in C. jejuni virulence, lipooligosaccharide (LOS) biosynthesis, and host colonization by targeted mutagenesis and complementation of the mutation. The cj1136 mutant showed a significant reduction in invasion into human intestinal epithelial cells compared to the wild-type strain 01/51. Invasion levels were partially restored on complementing the mutation. The inactivation of cj1136 resulted in the production of truncated LOS, while biosynthesis of a full-length LOS molecule was restored in the complemented strain. The cj1136 mutant showed an increase in sensitivity to the bile salts sodium taurocholate and sodium deoxycholate and significantly increased sensitivity to polymyxin B compared to the parental strain. Importantly, the ability of the mutant to colonize 1-day-old chicks was also significantly impaired. This study confirms that a putative galactosyltransferase encoded by cj1136 is involved in LOS biosynthesis and is important for C. jejuni virulence, as disruption of this gene and the resultant truncation of LOS affect both colonization in vivo and invasiveness in vitro. PMID:22508861

  16. Polyphosphate Kinase 2: A Novel Determinant of Stress Responses and Pathogenesis in Campylobacter jejuni

    PubMed Central

    Gangaiah, Dharanesh; Liu, Zhe; Arcos, Jesús; Kassem, Issmat I.; Sanad, Yasser; Torrelles, Jordi B.; Rajashekara, Gireesh

    2010-01-01

    Background Inorganic polyphosphate (poly P) plays an important role in stress tolerance and virulence in many bacteria. PPK1 is the principal enzyme involved in poly P synthesis, while PPK2 uses poly P to generate GTP, a signaling molecule that serves as an alternative energy source and a precursor for various physiological processes. Campylobacter jejuni, an important cause of foodborne gastroenteritis in humans, possesses homologs of both ppk1 and ppk2. ppk1 has been previously shown to impact the pathobiology of C. jejuni. Methodology/Principal Findings Here, we demonstrate for the first time that the deletion of ppk2 in C. jejuni resulted in a significant decrease in poly P-dependent GTP synthesis, while displaying an increased intracellular ATP:GTP ratio. The Δppk2 mutant exhibited a significant survival defect under osmotic, nutrient, aerobic, and antimicrobial stresses and displayed an enhanced ability to form static biofilms. However, the Δppk2 mutant was not defective in poly P and ppGpp synthesis suggesting that PPK2-mediated stress tolerance is not ppGpp-mediated. Importantly, the Δppk2 mutant was significantly attenuated in invasion and intracellular survival within human intestinal epithelial cells as well as in chicken colonization. Conclusions/Significance Taken together, we have highlighted the role of PPK2 as a novel pathogenicity determinant that is critical for C. jejuni survival, adaptation, and persistence in the host environments. PPK2 is absent in humans and animals; therefore, can serve as a novel target for therapeutic intervention of C. jejuni infections. PMID:20808906

  17. Survival of Campylobacter jejuni in mineral bottled water according to difference in mineral content: application of the Weibull model.

    PubMed

    Guillou, S; Leguerinel, I; Garrec, N; Renard, M A; Cappelier, J M; Federighi, M

    2008-04-01

    The aim of the study was to examine the hypothesis proposed by Evans et al. [2003. Hazards of healthy living: bottled water and salad vegetables as risk factors for Campylobacter infection. Emerg. Infect. Dis. 9(10), 1219-1225] that mineral bottled water accidentally contaminated by Campylobacter jejuni would represent a risk factor for Campylobacter infection. Culturability of C. jejuni cells inoculated in low- and high-mineral bottled water during storage at 4 degrees C in the dark was performed by surface plating and modelled using the Weibull model. The loss of C. jejuni culturability observed in all conditions tested was shown to be dependent on strain, preculture condition and water composition. Following inoculation of C. jejuni, the rapid loss of culturability was not correlated to complete cell death as the passage into embryonated eggs enabled recovery of cells from the viable but non-culturable state. In conclusion, the sanitary risk associated with contaminated bottled water cannot be excluded although it is presumably low. Culture conditions, strain and water type must be taken into account in the evaluation of the risk factors as they influence significantly Campylobacter survival in water.

  18. Creation of a Large Deletion Mutant of Campylobacter jejuni Reveals that the Lipooligosaccharide Gene Cluster Is Not Required for Viability▿

    PubMed Central

    Marsden, Gemma L.; Li, Jianjun; Everest, Paul H.; Lawson, Andrew J.; Ketley, Julian M.

    2009-01-01

    Deletion of the lipooligosaccharide biosynthesis region (Cj1132c to Cj1152c) from the genome of Campylobacter jejuni NCTC11168 shows that the core is not required for viability. The mutant was attenuated for growth and has increased sensitivity to antibiotics and detergents. Natural transformation and invasion of cultured host cells was abolished. PMID:19181814

  19. Untargeted metabolomic profiling of amphenicol-resistant Campylobacter jejuni by ultra-high-performance liquid chromatography-mass spectrometry.

    PubMed

    Li, Hui; Xia, Xi; Li, Xiaowei; Naren, Gaowa; Fu, Qin; Wang, Yang; Wu, Congming; Ding, Shuangyang; Zhang, Suxia; Jiang, Haiyang; Li, Jiancheng; Shen, Jianzhong

    2015-02-01

    Campylobacter jejuni, an important foodborne microorganism, poses severe and emergent threats to human health as antibiotic resistance becomes increasingly prevalent. The mechanisms of drug resistance are hard to decipher, and little is known at the metabolic level. Here we apply metabolomic profiling to discover metabolic changes associated with amphenicol (chloramphenicol and florfenicol) resistance mutations of Campylobacter jejuni. An optimized sample preparation method was combined with ultra-high-performance liquid chromatography-time-of-flight mass spectrometry (UHPLC-TOF/MS) and pattern recognition for the analysis of small-molecule biomarkers of drug resistance. UHPLC-triple quadrupole MS operated in multiple reaction monitoring mode was used for quantitative analysis of metabolic features from UHPLC-TOF/MS profiling. Up to 41 differential metabolites involved in glycerophospholipid metabolism, sphingolipid metabolism, and fatty acid metabolism were observed in a chloramphenicol-resistant mutant strain of Campylobacter jejuni. A panel of 40 features was identified in florfenicol-resistant mutants, demonstrating changes in glycerophospholipid metabolism, sphingolipid metabolism, and tryptophan metabolism. This study shows that the UHPLC-MS-based metabolomics platform is a promising and valuable tool to generate new insights into the drug-resistant mechanism of Campylobacter jejuni.

  20. Influence of temperature, oxygen and bacterial strain identity on the association of Campylobacter jejuni with Acanthamoeba castellanii.

    PubMed

    Baré, Julie; Sabbe, Koen; Huws, Sharon; Vercauteren, Dries; Braeckmans, Kevin; van Gremberghe, Ineke; Favoreel, Herman; Houf, Kurt

    2010-11-01

    Campylobacteriosis is the most frequently reported foodborne disease in the industrialized world, mainly through consumption of contaminated chicken meat. To date, no information is available on the primary infection sources of poultry. In this study, the ability of five Campylobacter jejuni strains with different invasion potential towards Caco-2 cells to survive and replicate in the protozoan Acanthamoeba castellanii was tested under simulated in situ conditions (i.e. chicken broiler houses). Results indicate that environmental conditions play a crucial role in C. jejuni-A. castellanii interactions. Co-culture in general did not result in an increase of either bacteria or amoebae. However, co-culture with Acanthamoeba did result in a delayed decline and an increased long-term survival of Campylobacter. Bacterial strain-specific effects were observed, with higher survival rates for low-invasive strains. The presence of C. jejuni in general did not affect A. castellanii viability, except at 37 °C under microaerobic conditions, where the presence of the reference and low-invasive Campylobacter strains resulted in a significant decline in amoebal viability. Confocal laser scanning microscopy revealed that intra-amoebal campylobacters were not always colocated with acidic organelles, suggesting potential bacterial interference with digestive processes. As Acanthamoeba enhances the persistence of C. jejuni, the presence of the amoeba in broiler house environments may have important implications for the ecology and epidemiology of this food pathogen. PMID:20722733

  1. Characterization of the Campylobacter jejuni cryptic plasmid pTIW94 recovered from wild birds in the southeastern United States

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The complete nucleotide sequence of a cryptic plasmid, pTIW94, recovered from several Campylobacter jejuni isolates from wild birds in the southeastern United States, was determined. All plasmids were circular molecules of 3865 nucleotides, with a G+C content of 31.0%, similar to that of Campylobac...

  2. Complete chemoenzymatic synthesis of the Forssman antigen using novel glycosyltransferases identified in Campylobacter jejuni and Pasteurella multocida

    Technology Transfer Automated Retrieval System (TEKTRAN)

    We have identified an alpha1,4-galactosyltransferase (CgtD) and a beta1,3-N-acetylgalactosaminyltransferase (CgtE) in the lipooligosaccharide (LOS) locus of Campylobacter jejuni LIO87. Strains that carry these genes may have the capability of synthesizing mimics of the P blood group antigens of the ...

  3. Campylobacter jejuni infection associated with unpasteurized milk and cheese--Kansas, 2007.

    PubMed

    2009-01-01

    On October 26, 2007, a family health clinic nurse informed the Kansas Department of Health and Environment (KDHE) that Campylobacter jejuni had been isolated from two ill persons from different families who were members of a closed community in a rural Kansas county. By October 29, 17 additional members of the community had reported gastrointestinal illness and visited the clinic within a week. All 19 persons reported consuming fresh cheese* on October 20 that was made the same day at a community fair from unpasteurized milk obtained from a local dairy. This report summarizes the findings of an investigation by KDHE and the local health department to determine the source and extent of the outbreak. Eating fresh cheese at the fair was the only exposure associated with illness (relative risk [RR] = 13.9). Of 101 persons who ate the cheese, 67 (66%) became ill. C. jejuni isolates from two ill persons had indistinguishable pulsed-field gel electrophoresis (PFGE) patterns, and the isolate from a third ill person was nearly identical to the other two. Although all samples of cheese tested negative for Campylobacter, results of the epidemiologic investigation found an association between illness and consumption of fresh cheese made from unpasteurized milk. To minimize the risk for illness associated with milkborne pathogens, unpasteurized milk and milk products should not be consumed.

  4. The mode of biofilm formation on smooth surfaces by Campylobacter jejuni.

    PubMed

    Moe, Kyaw Kyaw; Mimura, Junichiro; Ohnishi, Takahiro; Wake, Tomoya; Yamazaki, Wataru; Nakai, Masaaki; Misawa, Naoaki

    2010-04-01

    Many microorganisms produce extracellular polymers referred to collectively as "slime" or glycocalyx, and form biofilms on solid surfaces in natural ecosystems. Campylobacter jejuni, one of the most important foodborne pathogens, also has the ability to form biofilm on stainless steel, glass, or polyvinyl chloride in vitro. However, the issue of biofilm formation by Campylobacter species has not been extensively examined. The present study was performed to examine the mode of adhesion of C. jejuni to a smooth surface. When bacterial suspensions in Brucella broth were incubated in microplate wells with a glass coverslip, microcolonies 0.5~2 mm in diameter were formed on the coverslip within 2 hr from the start of incubation. These microcolonies gradually grew and formed a biofilm of net-like connections within 6 hr. Transmission electron microscopy indicated that massive amounts of extracellular material masked the cell surface, and this material bound ruthenium red, suggesting the presence of a polysaccharide moiety. Scanning electron microscopy indicated that the flagella acted as bridges, forming net-like connections between the organisms. To determine the genes associated with biofilm formation, aflagellate (flaA(-)) and flagellate but non-motile (motA(-)) mutants were constructed from strain 81-176 by natural transformation-mediated allelic exchange. The flaA(-) and motA(-) mutants did not form the biofilm exhibited by the wild-type strain. These findings suggest that flagella-mediated motility as well as flagella is required for biofilm formation in vitro.

  5. Survival of Campylobacter jejuni in co-culture with Acanthamoeba castellanii: role of amoeba-mediated depletion of dissolved oxygen.

    PubMed

    Bui, Xuan Thanh; Winding, Anne; Qvortrup, Klaus; Wolff, Anders; Bang, Dang Duong; Creuzenet, Carole

    2012-08-01

    Campylobacter jejuni is a major cause of infectious diarrhoea worldwide but relatively little is known about its ecology. In this study, we examined its interactions with Acanthamoeba castellanii, a protozoan suspected to serve as a reservoir for bacterial pathogens. We observed rapid degradation of intracellular C.jejuni in A.castellanii 5 h post gentamicin treatment at 25°C. Conversely, we found that A.castellanii promoted the extracellular growth of C.jejuni in co-cultures at 37°C in aerobic conditions. This growth-promoting effect did not require amoebae - bacteria contact. The growth rates observed with or without contact with amoeba were similar to those observed when C.jejuni was grown in microaerophilic conditions. Preconditioned media prepared with live or dead amoebae cultivated with or without C.jejuni did not promote the growth of C.jejuni in aerobic conditions. Interestingly, the dissolved oxygen levels of co-cultures with or without amoebae - bacteria contact were much lower than those observed with culture media or with C.jejuni alone incubated in aerobic conditions, and were comparable with levels obtained after 24 h of growth of C.jejuni under microaerophilic conditions. Our studies identified the depletion of dissolved oxygen by A.castellanii as the major contributor for the observed amoeba-mediated growth enhancement. PMID:22176643

  6. Application of real-time PCR for quantitative detection of Campylobacter jejuni in poultry, milk and environmental water.

    PubMed

    Yang, Chengbo; Jiang, Yuan; Huang, Kehe; Zhu, Changqing; Yin, Yulong

    2003-10-15

    Campylobacter jejuni is a leading human food-borne pathogen. The rapid and sensitive detection of C. jejuni is necessary for the maintenance of a safe food/water supply. In this article, we present a real-time polymerase chain reaction (PCR) assay for quantitative detection of C. jejuni in naturally contaminated poultry, milk and environmental samples without an enrichment step. The whole assay can be completed in 60 min with a detection limit of approximately 1 CFU. The standard curve correlation coefficient for the threshold cycle versus the copy number of initial C. jejuni cells was 0.988. To test the PCR system, a set of 300 frozen chicken meat samples, 300 milk samples and 300 water samples were screened for the presence of C. jejuni. 30.6% (92/300) of chicken meat samples, 27.3% (82/300) of milk samples, and 13.6% (41/300) of water samples tested positive for C. jejuni. This result indicated that the real-time PCR assay provides a specific, sensitive and rapid method for quantitative detection of C. jejuni. Moreover, it is concluded that retail chicken meat, raw milk and environmental water are commonly contaminated with C. jejuni and could serve as a potential risk for consumers in eastern China, especially if proper hygienic and cooking conditions are not maintained.

  7. Antimicrobial efficacy of lauric arginate against Campylobacter jejuni and spoilage organisms on chicken breast fillets.

    PubMed

    Nair, Divek V T; Nannapaneni, Rama; Kiess, Aaron; Mahmoud, Barakat; Sharma, Chander Shekhar

    2014-10-01

    The objectives of this study were to determine the antimicrobial efficacy of lauric arginate (LAE) against Campylobacter jejuni (in broth and on chicken breast fillets) and spoilage microorganisms (on chicken breast fillets). In vitro antimicrobial activity of LAE was determined by treating C. jejuni (in pure culture) with 0 (control), 50, 100, and 200 mg/L of LAE solutions at 4°C for 2 h. Inoculated chicken samples with C. jejuni were treated with 0, 200, and 400 mg/kg of LAE, packaged, and stored at 4°C for 7 d for determining the efficacy of LAE against C. jejuni on meat. Noninoculated skinless chicken breast fillet samples were treated with 0, 200, and 400 mg/kg of LAE and were used for analysis of LAE treatments on growth of mesophilic and psychrotrophic organisms on d 0, 3, 9, and 14 during storage at 4°C. Lauric arginate was highly effective against C. jejuniin vitro with no detectable survivors. Lauric arginate significantly (P ≤ 0.05) reduced C. jejuni counts on chicken breast fillets with 200 and 400 mg/kg treatments. Lauric arginate at 400 mg/L gave a maximum reduction of ~1.5 log cfu/g of C. jejuni during 7 d of storage at 4°C without any change in pH of meat. Treating chicken breast fillets with 400 mg/kg of LAE caused 2.3 log cfu/g reduction of psychrotrophs (P ≤ 0.05) compared with the control on d 0 of storage. However, no difference existed (P ≥ 0.05) in the growth of psychrotrophs on chicken breast fillets after treatment with 200 and 400 mg/kg of LAE compared with the control after 3 d. The LAE treatments had no effect (P ≥ 0.05) on growth of mesophilic organisms. The results of the study indicated that LAE is effective in reducing C. jejuni on chicken breast fillets.

  8. Novel Immunomodulatory Flagellin-Like Protein FlaC in Campylobacter jejuni and Other Campylobacterales

    PubMed Central

    Faber, Eugenia; Gripp, Eugenia; Maurischat, Sven; Kaspers, Bernd; Tedin, Karsten; Menz, Sarah; Zuraw, Aleksandra; Kershaw, Olivia; Yang, Ines; Rautenschlein, Silke

    2015-01-01

    ABSTRACT The human diarrheal pathogens Campylobacter jejuni and Campylobacter coli interfere with host innate immune signaling by different means, and their flagellins, FlaA and FlaB, have a low intrinsic property to activate the innate immune receptor Toll-like receptor 5 (TLR5). We have investigated here the hypothesis that the unusual secreted, flagellin-like molecule FlaC present in C. jejuni, C. coli, and other Campylobacterales might activate cells via TLR5 and interact with TLR5. FlaC shows striking sequence identity in its D1 domains to TLR5-activating flagellins of other bacteria, such as Salmonella, but not to nonstimulating Campylobacter flagellins. We overexpressed and purified FlaC and tested its immunostimulatory properties on cells of human and chicken origin. Treatment of cells with highly purified FlaC resulted in p38 activation. FlaC directly interacted with TLR5. Preincubation with FlaC decreased the responsiveness of chicken and human macrophage-like cells toward the bacterial TLR4 agonist lipopolysaccharide (LPS), suggesting that FlaC mediates cross-tolerance. C. jejuni flaC mutants induced an increase of cell responses in comparison to those of the wild type, which was suppressed by genetic complementation. Supplementing excess purified FlaC likewise reduced the cellular response to C. jejuni. In vivo, the administration of ultrapure FlaC led to a decrease in cecal interleukin 1β (IL-1β) expression and a significant change of the cecal microbiota in chickens. We propose that Campylobacter spp. have evolved a novel type of secreted immunostimulatory flagellin-like effector in order to specifically modulate host responses, for example toward other pattern recognition receptor (PRR) ligands, such as LPS. IMPORTANCE Flagellins not only are important for bacterial motility but are major bacterial proteins that can modulate host responses via Toll-like receptor 5 (TLR5) or other pattern recognition receptors. Campylobacterales colonizing the

  9. Novel Immunomodulatory Flagellin-Like Protein FlaC in Campylobacter jejuni and Other Campylobacterales.

    PubMed

    Faber, Eugenia; Gripp, Eugenia; Maurischat, Sven; Kaspers, Bernd; Tedin, Karsten; Menz, Sarah; Zuraw, Aleksandra; Kershaw, Olivia; Yang, Ines; Rautenschlein, Silke; Josenhans, Christine

    2016-01-01

    The human diarrheal pathogens Campylobacter jejuni and Campylobacter coli interfere with host innate immune signaling by different means, and their flagellins, FlaA and FlaB, have a low intrinsic property to activate the innate immune receptor Toll-like receptor 5 (TLR5). We have investigated here the hypothesis that the unusual secreted, flagellin-like molecule FlaC present in C. jejuni, C. coli, and other Campylobacterales might activate cells via TLR5 and interact with TLR5. FlaC shows striking sequence identity in its D1 domains to TLR5-activating flagellins of other bacteria, such as Salmonella, but not to nonstimulating Campylobacter flagellins. We overexpressed and purified FlaC and tested its immunostimulatory properties on cells of human and chicken origin. Treatment of cells with highly purified FlaC resulted in p38 activation. FlaC directly interacted with TLR5. Preincubation with FlaC decreased the responsiveness of chicken and human macrophage-like cells toward the bacterial TLR4 agonist lipopolysaccharide (LPS), suggesting that FlaC mediates cross-tolerance. C. jejuni flaC mutants induced an increase of cell responses in comparison to those of the wild type, which was suppressed by genetic complementation. Supplementing excess purified FlaC likewise reduced the cellular response to C. jejuni. In vivo, the administration of ultrapure FlaC led to a decrease in cecal interleukin 1β (IL-1β) expression and a significant change of the cecal microbiota in chickens. We propose that Campylobacter spp. have evolved a novel type of secreted immunostimulatory flagellin-like effector in order to specifically modulate host responses, for example toward other pattern recognition receptor (PRR) ligands, such as LPS. IMPORTANCE Flagellins not only are important for bacterial motility but are major bacterial proteins that can modulate host responses via Toll-like receptor 5 (TLR5) or other pattern recognition receptors. Campylobacterales colonizing the intestinal

  10. The Fecal Microbial Communities of Dairy Cattle Shedding Shiga Toxin-Producing Escherichia coli or Campylobacter jejuni.

    PubMed

    Dong, Hee-Jin; Kim, Woohyun; An, Jae-Uk; Kim, Junhyung; Cho, Seongbeom

    2016-09-01

    Cattle are a natural reservoir of Shiga toxin-producing Escherichia coli (STEC) and have recently been recognized as a major source of Campylobacter jejuni contamination. While several factors are known to be associated with bacterial colonization, the underlying microbial factors have not been clarified. In this study, we characterized the fecal microbiota of dairy cattle (n = 24) using next-generation sequencing to elucidate the intestinal bacterial communities and the microbial diversity in relation to the presence of the foodborne pathogens STEC and C. jejuni (STEC-positive samples, n = 9; STEC-negative samples, n = 15; C. jejuni-positive samples, n = 9; and C. jejuni-negative samples, n = 15). While no significant differences were observed in alpha diversity between STEC-positive and STEC-negative samples, a high diversity index was observed in C. jejuni-positive samples compared to C. jejuni-negative samples. Nine phyla, 13 classes, 18 orders, 47 families, 148 genera, and 261 species were found to be the core microbiota in dairy cattle, covering 80.0-100.0% of the fecal microbial community. Diverse microbial communities were observed between cattle shedding foodborne pathogens and nonshedding cattle. C. jejuni-positive cattle had a higher relative abundance of Bacteroidetes (p = 0.035) and a lower relative abundance of Firmicutes (p = 0.035) compared to C. jejuni-negative cattle. In addition, while the relative abundance of 2 and 6 genera was significantly higher in cattle-shedding STEC and C. jejuni, respectively, the relative abundance of 3 genera was lower in both STEC- and C. jejuni-negative cattle. Our findings provide fundamental information on the bacterial ecology in cattle feces and might be useful in developing strategies to reduce STEC or C. jejuni shedding in dairy cattle, thereby reducing the incidence of STEC infection and campylobacteriosis in humans. PMID:27385033

  11. Antimicrobial activity of copper surfaces against suspensions of Salmonella enterica and Campylobacter jejuni

    PubMed Central

    Faúndez, Gustavo; Troncoso, Miriam; Navarrete, Paola; Figueroa, Guillermo

    2004-01-01

    Background Salmonella enterica and Campylobacter jejuni are amongst the more prevalent bacterial pathogens that cause foodborne diseases. These microorganisms are common contaminants of poultry and poultry products. This study was aimed to evaluate the antibacterial activity of metallic copper surfaces on these important enteropathogens, and to determine the potential acquisition of copper by food exposed to this metal. Results The antibacterial activity of copper surfaces was evaluated overlying them with suspensions of 106 CFU/ml of S. enterica and C. jejuni. Bacterial counts obtained after 0, 2, 4 and 8 hours at 10°C and 25°C were compared with those obtained in stainless steel and a synthetic polymer as control surfaces. The results showed that when these enteropathogens were kept in contact with copper a significant antibacterial activity was noted, on the contrary when the same load of pathogen suspensions were tested over the control surfaces it was found that the bacterial counts remained unchanged or even increased with time. The potential acquisition of copper by food exposed to this surface was also evaluated. Meat exposed for one hour to a copper surface adsorbed residual copper in a time dependant manner. Conclusions These results shows that metallic copper surfaces have an antibacterial activity against S. enterica and C. jejuni and suggest its potential application as an inhibitory agent in the various stages of the food processing operations. PMID:15119960

  12. 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

  13. Active migration into the subcellular space precedes Campylobacter jejuni invasion of epithelial cells.

    PubMed

    van Alphen, Lieke B; Bleumink-Pluym, Nancy M C; Rochat, Klazina D; van Balkom, Bas W M; Wösten, Marc M S M; van Putten, Jos P M

    2008-01-01

    The bacterial pathogen Campylobacter jejuni invades mucosal cells via largely undefined and rather inefficient (0.01-2 bacteria per cell) mechanisms. Here we report a novel, highly efficient C. jejuni infection pathway resulting in 10-15 intracellular bacteria per cell within 3 h of infection. Electron microscopy, pulse-chase infection assays and time-lapse multiphoton laser confocal microscopy demonstrated that the mechanism involved active and rapid migration of the pathogen into the subcellular space (termed 'subvasion'), followed by bacterial entry ('invasion') at the cell basis. Efficient subvasion was maximal after repeated rounds of selection for the subvasive phenotype. Targeted mutagenesis indicated that the CadF, JlpA or PEB1 adhesins were not required. Dissection of the selected and parental phenotypes by SDS-PAGE yielded comparable capsule polysaccharide and lipooligosaccharide profiles. Proteomics revealed reduced amounts of the chemotaxis protein CheW for the subvasive phenotype. Swarming assays confirmed that the selected phenotype exhibited altered migration behaviour. Introduction of a plasmid carrying chemotaxis genes into the subvasive strain yielded wild-type subvasion levels and migration behaviour. These results indicate that alterations in the bacterial migration machinery enable C. jejuni to actively penetrate the subcellular space and gain access to the cell interior with unprecedented efficiency. PMID:18052944

  14. Molecular Mechanisms and Potential Clinical Applications of Campylobacter jejuni Cytolethal Distending Toxin

    PubMed Central

    Lai, Cheng-Kuo; Chen, Yu-An; Lin, Chun-Jung; Lin, Hwai-Jeng; Kao, Min-Chuan; Huang, Mei-Zi; Lin, Yu-Hsin; Chiang-Ni, Chuan; Chen, Chih-Jung; Lo, U-Ging; Lin, Li-Chiung; Lin, Ho; Hsieh, Jer-Tsong; Lai, Chih-Ho

    2016-01-01

    Cytolethal distending toxin (CDT), a genotoxin produced by Campylobacter jejuni, is composed of three subunits: CdtA, CdtB, and CdtC. CdtB is a DNase that causes DNA double-strand breaks (DSB) in the nucleus resulting in cell cycle arrest at the G2/M stage and apoptosis. CdtA and CdtC bind to cholesterol-rich microdomains on the cytoplasmic membrane, a process required for the delivery of CdtB to cells. Although a unique motif associated with cholesterol-binding activity has been identified in other pathogens, the mechanism underlying the interaction between the CdtA and CdtC subunits and membrane cholesterol remains unclear. Also, the processes of cell uptake and delivery of CdtB in host cells and the translocation of CdtB into the nucleus are only partially understood. In this review, we focus on the underlying relationship among CDT, membrane cholesterol, and the intracellular trafficking pathway as a unique mechanism for C. jejuni-induced pathogenesis. Moreover, we discuss the clinical aspects of a possible therapeutic application of CDT in cancer therapy. Understanding the molecular mechanism of CDT-host interactions may provide insights into novel strategies to control C. jejuni infection and the development of potential clinical applications of CDT. PMID:26904508

  15. Colonization of chicken flocks by Campylobacter jejuni in multiple farms in Japan.

    PubMed

    Yano, S; Kira, T; Morishita, Y; Ishihara, K; Asai, T; Iwata, T; Akiba, M; Murase, T

    2013-02-01

    Campylobacter jejuni was monitored in 4 chicken farms during the period 2003 to 2006 to elucidate the mechanisms of transmission. Three farms (1 to 3), located at least 14 km from each other, belonged to an integrated poultry company, which also provided the farms with day-old chicks from several hatcheries as well as chicken feed. Another farm (4), which belonged to a different company, was located 270 m from farm 1. A total of 206 C. jejuni isolates obtained from the 4 farms were classified into 10 flaA-based RFLP types. Identical RFLP types were found in isolates obtained from chickens originating from multiple hatcheries and reared in different chicken houses on individual farms. Flocks were colonized by strains with 1 or 2 RFLP types in each production cycle, sometimes differing between cycles. Identical RFLP types were found in isolates obtained from the environment around the chicken houses. Using multilocus sequence typing, strains with different RFLP types could be distinguished from each other. Identical RFLP and multilocus sequence typing profiles were found in isolates obtained from farms 1 and 4, and from farms 1 and 2. These results suggest that C. jejuni in these farms comes from common sources external to the farms, even if the farms belong to different companies and obtain chicks from different suppliers.

  16. Antibacterial activity of three medicinal Thai plants against Campylobacter jejuni and other foodborne pathogens.

    PubMed

    Dholvitayakhun, Achara; Cushnie, T P Tim; Trachoo, Nathanon

    2012-01-01

    Leaves of Adenanthera pavonina, Moringa oleifera and Annona squamosa are used in traditional Thai medicine to treat dysentery and other diseases. This study investigated the antibacterial activity of these plants against six species of foodborne pathogen. Methods and solvents employed to extract active constituents were optimised using the disc diffusion assay. Phytochemical analysis of the optimised extracts was performed by thin layer chromatography (TLC). Minimum inhibitory concentrations (MICs) and minimum bactericidal concentrations (MBCs) were determined by broth microdilution. A. pavonina contained flavonoids, terpines and tannins, and was the most active extract against Campylobacter jejuni, inhibiting growth at 62.5-125 µg mL(-1). The A. squamosa extract contained flavonoids, terpines, tannins and alkaloids, and had the broadest spectrum of antibacterial activity, inhibiting Bacillus cereus, Listeria monocytogenes, Staphylococcus aureus and C. jejuni between 62.5 and 500 µg mL(-1). MBCs were 2- to 4-fold higher than MICs against C. jejuni and B. cereus, suggesting the extracts are bactericidal against these species. Negligible activity was detected from M. oleifera. The data presented here show that A. pavonina and A. squamosa could potentially be used in modern applications aimed at the treatment or prevention of foodborne diseases. PMID:21878033

  17. Antibacterial activity of three medicinal Thai plants against Campylobacter jejuni and other foodborne pathogens.

    PubMed

    Dholvitayakhun, Achara; Cushnie, T P Tim; Trachoo, Nathanon

    2012-01-01

    Leaves of Adenanthera pavonina, Moringa oleifera and Annona squamosa are used in traditional Thai medicine to treat dysentery and other diseases. This study investigated the antibacterial activity of these plants against six species of foodborne pathogen. Methods and solvents employed to extract active constituents were optimised using the disc diffusion assay. Phytochemical analysis of the optimised extracts was performed by thin layer chromatography (TLC). Minimum inhibitory concentrations (MICs) and minimum bactericidal concentrations (MBCs) were determined by broth microdilution. A. pavonina contained flavonoids, terpines and tannins, and was the most active extract against Campylobacter jejuni, inhibiting growth at 62.5-125 µg mL(-1). The A. squamosa extract contained flavonoids, terpines, tannins and alkaloids, and had the broadest spectrum of antibacterial activity, inhibiting Bacillus cereus, Listeria monocytogenes, Staphylococcus aureus and C. jejuni between 62.5 and 500 µg mL(-1). MBCs were 2- to 4-fold higher than MICs against C. jejuni and B. cereus, suggesting the extracts are bactericidal against these species. Negligible activity was detected from M. oleifera. The data presented here show that A. pavonina and A. squamosa could potentially be used in modern applications aimed at the treatment or prevention of foodborne diseases.

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

    PubMed

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

    2012-07-19

    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.

  19. Effect of human isolated probiotic bacteria on preventing Campylobacter jejuni colonization of poultry.

    PubMed

    Cean, Ada; Stef, Lavinia; Simiz, Eliza; Julean, Calin; Dumitrescu, Gabi; Vasile, Aida; Pet, Elena; Drinceanu, Dan; Corcionivoschi, Nicolae

    2015-02-01

    This study was performed in order to determine whether human isolated probiotic bacteria can be effective in reducing Campylobacter jejuni infection of chicken intestinal cells, in vitro, and in decreasing its colonization abilities within the chicken gut. Our results show that the probiotic strains Lactobacillus paracasei J. R, L. rhamnosus 15b, L. lactis Y, and L. lactis FOa had a significant effect on C. jejuni invasion of chicken primary cells, with the strongest inhibitory effect detected when a combination of four was administered. In regard to the in vivo effect, using all four strains in one combination prevented mucus colonization in the duodenum and cecum. Moreover, the pathogen load in the lumen of these two compartments was significantly reduced. When probiotics were introduced during the early growth period, the presence of the pathogen in feces was increased (p>0.05), but when they were given during the last week of growth, there was no significant effect. In conclusion, our data indicate that these four new probiotic strains are able to cause modifications in the chicken intestinal mucosa and can reduce the ability of C. jejuni to invade, in vitro, and to colonize, in vivo. These probiotics are now proven to be effective even when introduced in broiler's feed 7 days before slaughter, which makes them cost-effective for the producers.

  20. Evolution and comparative genomics of Campylobacter jejuni ST-677 clonal complex.

    PubMed

    Kivistö, Rauni I; Kovanen, Sara; Skarp-de Haan, Astrid; Schott, Thomas; Rahkio, Marjatta; Rossi, Mirko; Hänninen, Marja-Liisa

    2014-09-04

    Campylobacter is the most common bacterial cause of gastroenteritis in the European Union with over 200,000 laboratory-confirmed cases reported annually. This is the first study to describe findings related to comparative genomics analyses of the sequence type (ST)-677 clonal complex (CC), a Campylobacter jejuni lineage associated with bacteremia cases in humans. We performed whole-genome sequencing, using Illumina HiSeq sequencing technology, on five related ST-677 CC isolates from two chicken farms to identify microevolution taking place at the farms. Our further aim was to identify novel putative virulence determinants from the ST-677 CC genomes. For this purpose, clinical isolates of the same CC were included in comparative genomic analyses against well-known reference strains of C. jejuni. Overall, the ST-677 CC was recognized as a highly clonal lineage with relatively small differences between the genomes. Among the farm isolates differences were identified mainly in the lengths of the homopolymeric tracts in genes related to the capsule, lipo-oligosaccharide, and flagella. We identified genomic features shared with C. jejuni subsp. doylei, which has also been shown to be associated with bacteremia in humans. These included the degradation of the cytolethal distending toxin operon and similarities between the capsular polysaccharide biosynthesis loci. The phase-variable GDP-mannose 4,6-dehydratase (EC 4.2.1.47) (wcbK, CAMP1649), associated with the capsular polysaccharide biosynthesis locus, may play a central role in ST-677 CC conferring acid and serum resistance during different stages of infection. Homology-based searches revealed several additional novel features and characteristics, including two putative type Vb secretion systems and a novel restriction modification/methyltransferase gene cluster, putatively associated with pathogenesis and niche adaptation.

  1. Antibacterial effect of trans-cinnamaldehyde, eugenol, carvacrol, and thymol on Salmonella Enteritidis and Campylobacter jejuni in chicken cecal contents in vitro

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Salmonella Enteritidis and Campylobacter jejuni are two major food-borne pathogens that are transmitted through poultry products. These pathogens colonize the chicken cecum leading, to contamination of carcasses during slaughter and subsequent processing operations. We investigated the antimicrobial...

  2. Production of a bacteriocin by a poultry derived Campylobacter jejuni isolate with antimicrobial activity against Clostridium perfringens and other Gram positive bacteria.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    We have purified a bacteriocin peptide (termed CUV-3), produced by a poultry cecal isolate of Campylobacter jejuni (strain CUV-3) with inhibitory activity against Gram positive bacteria including Clostridium perfringens (38 strains), Staphylococcus aureus, Staphylococcus epidermidis and Listeria mon...

  3. Pancreatic Amylase Is an Environmental Signal for Regulation of Biofilm Formation and Host Interaction in Campylobacter jejuni

    PubMed Central

    Jowiya, Waheed; Brunner, Katja; Abouelhadid, Sherif; Hussain, Haitham A.; Nair, Sean P.; Sadiq, Sohaib; Williams, Lisa K.; Trantham, Emma K.; Stephenson, Holly; Wren, Brendan W.; Bajaj-Elliott, Mona; Cogan, Tristan A.; Laws, Andrew P.; Wade, Jim; Dorrell, Nick

    2015-01-01

    Campylobacter jejuni is a commensal bacterium in the intestines of animals and birds and a major cause of food-borne gastroenteritis in humans worldwide. Here we show that exposure to pancreatic amylase leads to secretion of an α-dextran by C. jejuni and that a secreted protease, Cj0511, is required. Exposure of C. jejuni to pancreatic amylase promotes biofilm formation in vitro, increases interaction with human epithelial cell lines, increases virulence in the Galleria mellonella infection model, and promotes colonization of the chicken ileum. We also show that exposure to pancreatic amylase protects C. jejuni from stress conditions in vitro, suggesting that the induced α-dextran may be important during transmission between hosts. This is the first evidence that pancreatic amylase functions as an interkingdom signal in an enteric microorganism. PMID:26438798

  4. Pancreatic amylase is an environmental signal for regulation of biofilm formation and host interaction in Campylobacter jejuni.

    PubMed

    Jowiya, Waheed; Brunner, Katja; Abouelhadid, Sherif; Hussain, Haitham A; Nair, Sean P; Sadiq, Sohaib; Williams, Lisa K; Trantham, Emma K; Stephenson, Holly; Wren, Brendan W; Bajaj-Elliott, Mona; Cogan, Tristan A; Laws, Andrew P; Wade, Jim; Dorrell, Nick; Allan, Elaine

    2015-12-01

    Campylobacter jejuni is a commensal bacterium in the intestines of animals and birds and a major cause of food-borne gastroenteritis in humans worldwide. Here we show that exposure to pancreatic amylase leads to secretion of an α-dextran by C. jejuni and that a secreted protease, Cj0511, is required. Exposure of C. jejuni to pancreatic amylase promotes biofilm formation in vitro, increases interaction with human epithelial cell lines, increases virulence in the Galleria mellonella infection model, and promotes colonization of the chicken ileum. We also show that exposure to pancreatic amylase protects C. jejuni from stress conditions in vitro, suggesting that the induced α-dextran may be important during transmission between hosts. This is the first evidence that pancreatic amylase functions as an interkingdom signal in an enteric microorganism.

  5. Comparison of Campylobacter jejuni pulsotypes isolated from humans and poultry in Split and Dalmatia County, Croatia.

    PubMed

    Kovačić, Ana; Carev, Merica; Tripković, Ingrid; Srečec, Siniša; Siško-Kraljević, Katarina

    2015-01-01

    Consumption of poultry is considered to be an important source of human infection with Campylobacter. In the period from 2008 to 2010, 50 isolates of Campylobacter jejuni from human faeces were analysed and compared with 61 isolates from poultry by pulsed field gel electrophoresis using SmaI and KpnI. Based on the analysis of SmaI macrorestriction profiles, 86 isolates (77.5 %) were assigned to 15 S clusters: 31 (62 %) from humans and 55 from poultry (90.2 %). Altogether 21 isolates (19 %) exhibited macrorestriction profiles common to both humans and poultry after restriction with SmaI and KpnI. A total of five identical pulsotypes were isolated from both poultry and patients and one of them appeared in eight different locations in the time interval of one year. These results indicate that poultry could be an important source of Campylobacter infection in Split and Dalmatia County which is the biggest County in Croatia and the most important tourist destination.

  6. Effects of Lipooligosaccharide Inner Core Truncation on Bile Resistance and Chick Colonization by Campylobacter jejuni

    PubMed Central

    Iwata, Taketoshi; Chiku, Kazuhiro; Amano, Ken-ichi; Kusumoto, Masahiro; Ohnishi-Kameyama, Mayumi; Ono, Hiroshi; Akiba, Masato

    2013-01-01

    Campylobacter jejuni is the most common bacterium that causes diarrhea worldwide, and chickens are considered the main reservoir of this pathogen. This study investigated the effects of serial truncation of lipooligosaccharide (LOS), a major component of the outer membrane of C. jejuni, on its bile resistance and intestinal colonization ability in chickens. Genes encoding manno-heptose synthetases or glycosyltransferases were inactivated to generate isogenic mutants. Serial truncation of the LOS core oligosaccharide caused a stepwise increase in susceptibilities of two C. jejuni strains, NCTC 11168 and 81-176, to bile acids. Inactivation of hldE, hldD, or waaC caused severe truncation of the core oligosaccharide, which greatly increased the susceptibility to bile acids. Both wild-type strains grew normally in chicken intestinal extracts, whereas the mutants with severe oligosaccharide truncation were not detected 12 h after inoculation. These mutants attained viable bacterial counts in the bile acid-free extracts 24 h after inoculation. The wild-type strain 11-164 was present in the cecal contents at >107 CFU/g on 5 days after challenge infection and after this time period, whereas its hldD mutant was present at <103 CFU/g throughout the experimental period. Trans-complementation of the hldD mutant with the wild-type hldD allele completely restored the in vivo colonization level to that of the wild-type strain. Mutants with a shorter LOS had higher hydrophobicities. Thus, the length of the LOS core oligosaccharide affected the surface hydrophobicity and bile resistance of C. jejuni as well as its ability to colonize chicken intestines. PMID:23437265

  7. EptC of Campylobacter jejuni mediates phenotypes involved in host interactions and virulence.

    PubMed

    Cullen, Thomas W; O'Brien, John P; Hendrixson, David R; Giles, David K; Hobb, Rhonda I; Thompson, Stuart A; Brodbelt, Jennifer S; Trent, M Stephen

    2013-02-01

    Campylobacter jejuni is a natural commensal of the avian intestinal tract. However, the bacterium is also the leading cause of acute bacterial diarrhea worldwide and is implicated in development of Guillain-Barré syndrome. Like many bacterial pathogens, C. jejuni assembles complex surface structures that interface with the surrounding environment and are involved in pathogenesis. Recent work in C. jejuni identified a gene encoding a novel phosphoethanolamine (pEtN) transferase, EptC (Cj0256), that plays a promiscuous role in modifying the flagellar rod protein, FlgG; the lipid A domain of lipooligosaccharide (LOS); and several N-linked glycans. In this work, we report that EptC catalyzes the addition of pEtN to the first heptose sugar of the inner core oligosaccharide of LOS, a fourth enzymatic target. We also examine the role pEtN modification plays in circumventing detection and/or killing by host defenses. Specifically, we show that modification of C. jejuni lipid A with pEtN results in increased recognition by the human Toll-like receptor 4-myeloid differentiation factor 2 (hTLR4-MD2) complex, along with providing resistance to relevant mammalian and avian antimicrobial peptides (i.e., defensins). We also confirm the inability of aberrant forms of LOS to activate Toll-like receptor 2 (TLR2). Most exciting, we demonstrate that strains lacking eptC show decreased commensal colonization of chick ceca and reduced colonization of BALB/cByJ mice compared to wild-type strains. Our results indicate that modification of surface structures with pEtN by EptC is key to its ability to promote commensalism in an avian host and to survive in the mammalian gastrointestinal environment.

  8. Isolation and enumeration of Campylobacter jejuni from poultry products by a selective enrichment method.

    PubMed

    Wesley, R D; Swaminathan, B; Stadelman, W J

    1983-11-01

    A direct selective enrichment procedure was developed for the isolation of Campylobacter jejuni from poultry products. The selective enrichment medium (ATB) consisted of (per liter) tryptose (20 g), yeast extract (2.5 g), sodium chloride (5 g), FBP supplement (ferrous sulfate [0.25 g], sodium metabisulfite [0.25 g], sodium pyruvate [0.25 g]), bicine (10 g), and agar (1 g). Hematin solution (6.25 ml; prepared by dissolving 0.032 g of bovine hemin in 10 ml of 0.15 N sodium hydroxide solution and autoclaving at 0.35 kg/cm2 for 30 min), rifampin (25 mg), cefsulodin (6.25 mg), and polymyxin B sulfate (20,000 IU) were added after the medium was sterilized. The pH was adjusted to 8.0. Samples were enriched in the above medium at 42 degrees C for 48 h under an atmosphere of 5% O2, 10% CO2, and 85% N2. Enrichment cultures were streaked on a plating medium composed of Brucella agar, hematin solution, FBP supplement, and the above antibiotics. Plates were incubated under the same conditions as above. Suspect colonies from the plates were confirmed to be C. jejuni by morphological examination, growth characteristics, and biochemical tests. The above method yielded 25 isolates of C. jejuni from 50 samples of retail cut-up chicken and chicken parts, whereas a more complex method involving filtration, centrifugation, selective enrichment under a flowing atmosphere, and membrane filtration yielded only 6 positives from the same samples. The new isolation procedure was particularly effective in isolating C. jejuni in the presence of large numbers of Pseudomonas aeruginosa.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:6651294

  9. 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

    Caballero, Moisés; Rivera, Isabel; Jara, Luis M; Ulloa-Stanojlovic, Francisco M; Shiva, Carlos

    2015-01-01

    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. coli were 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.

  10. Genetic diversity and antibiotic resistance profiles of Campylobacter jejuni isolates from poultry and humans in Turkey.

    PubMed

    Abay, Secil; Kayman, Tuba; Otlu, Baris; Hizlisoy, Harun; Aydin, Fuat; Ertas, Nurhan

    2014-05-16

    In this study, the investigation of clonal relations between human and poultry Campylobacter jejuni isolates and the determination of susceptibilities of isolates to various antibiotics were aimed. A total of 200 C. jejuni isolates concurrently obtained from 100 chicken carcasses and 100 humans were genotyped by the Pulsed-Field Gel Electrophoresis (PFGE) and automated Repetitive Extragenic Palindromic PCR (Rep-PCR, DiversiLab system) methods and were tested for their susceptibility to six antibiotics with disk diffusion method. The minimum inhibitory concentration (MIC) values of ciprofloxacin (CI), enrofloxacin (EF) and erythromycin (EM) were evaluated by E-test. By using PFGE 174 of (87.0%) the isolates were able to be typed. The clonally related strains were placed in 35 different clusters and 115 different genotypes were obtained. All of the two hundred isolates could be typed by using Rep-PCR and were divided into 133 different genotypes. One hundred and fourteen clonally related isolates (57.0%) were included in 47 clusters. In disk diffusion test, while the susceptibility rates of AMC and S to human and chicken derived C. jejuni isolates were 84.0%-96.0% and 96.0%-98.0%, respectively, all isolates were susceptible to gentamicin. The resistance rates of human isolates to AMP, NA and TE were detected as 44.0%, 84.0% and 38.0% of the resistances of chicken isolates to these antibiotics were 34.0%, 95.0% and 56.0%, respectively. The MIC values of human and chicken isolates to CI, EF and EM were detected as 81.0-93.0%, 85.0-88.0% and 6.0-7.0%, respectively. The clonal proximity rates were detected between human and poultry origin C. jejuni isolates. The discriminatory power of PFGE and Rep-PCR was similar, with Simpson's diversity indexes of 0.993 and 0.995, respectively. Concordance of the two methods as determined by Adjusted Rand coefficient was 0.198 which showed the low congruence between Rep-PCR and PFGE. High rates of quinolone resistance were detected in

  11. 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

  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

    2016-01-01

    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

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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...

  14. Detection of Campylobacter jejuni and Campylobacter coli in chicken meat samples by real-time nucleic acid sequence-based amplification with molecular beacons.

    PubMed

    Churruca, E; Girbau, C; Martínez, I; Mateo, E; Alonso, R; Fernández-Astorga, A

    2007-06-10

    A nucleic acid sequence-based amplification (NASBA) assay based on molecular beacons was used for real-time detection of Campylobacter jejuni and Campylobacter coli in samples of chicken meat. A set of specific primers and beacon probe were designed to target the 16S rRNA of both species. The real-time NASBA protocol including the RNA isolation was valid for both of the cell suspensions in buffered saline and the artificially contaminated chicken meat samples. The presence of rRNA could be correlated with cellular viability, following inactivation of the bacteria by heating, in inoculated chicken meat samples but not in RNase-free cell suspensions.

  15. 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

  16. Direct microscopic observation of viability of Campylobacter jejuni on chicken skin treated with selected chemical sanitizing agents.

    PubMed

    Chantarapanont, Walairut; Berrang, Mark E; Frank, Joseph F

    2004-06-01

    The objective of this research was to determine the effect of chlorine, acidified sodium chlorite, and peracetic acid treatments on viable Campylobacter jejuni located at various depths within follicles or folds of chicken skin. Chicken skin was inoculated with C. jejuni transformed with P(c)gfp plasmid (GFP-Campylobacter), which also codes for kanamycin resistance. Effectiveness of sanitizer treatments was determined by plate count. C. jejuni were also observed on chicken skin by confocal scanning laser microscopy, whereby viable and nonviable cells were differentiated by their ability to take up staining with 5-cyano-2,3-ditolyl tetrazolium chloride. Sodium hypochlorite, peracetic acid, and acidified sodium chlorite were each applied at 40 or 100 ppm for 2 or 15 min. Each sanitizer resulted in approximately a 1-log decrease (CFU) when used at 100 ppm for 15 min and no significant decrease when used at 40 ppm for 2 min. Numbers of viable cells observed on the skin by direct microscopic count were similar to numbers obtained by plate count. Although viable counts decreased with sanitizer treatments, the total number of Campylobacter cells (live plus dead) attached to the skin remained unchanged. After each chemical treatment, viable C. jejuni were observed at depths of 0 to 10, 11 to 20, and 21 to 30 microm in folds or follicles of chicken skin. Most of the C. jejuni that survived treatment were located at 0 to 10 microm depth, which is where most of the viable cells were located before treatment. The inability of chemical sanitizers to effectively eliminate C. jejuni on chicken skin does not appear to be a result of protection by location in feather follicles or other depressions in the skin. PMID:15222541

  17. Antimicrobial activity of essential oils and five terpenoid compounds against Campylobacter jejuni in pure and mixed culture experiments.

    PubMed

    Kurekci, Cemil; Padmanabha, Jagadish; Bishop-Hurley, Sharon L; Hassan, Errol; Al Jassim, Rafat A M; McSweeney, Christopher S

    2013-09-16

    The aim of this study was to examine the antimicrobial potential of three essential oils (EOs: tea tree oil, lemon myrtle oil and Leptospermum oil), five terpenoid compounds (α-bisabolol, α-terpinene, cineole, nerolidol and terpinen-4-ol) and polyphenol against two strains of Campylobacter jejuni (ACM 3393 and the poultry isolate C338), Campylobacter coli and other Gram negative and Gram positive bacteria. Different formulations of neem oil (Azadirachta indica) with these compounds were also tested for synergistic interaction against all organisms. Antimicrobial activity was determined by the use of disc diffusion and broth dilution assays. All EOs tested were found to have strong antimicrobial activity against Campylobacter spp. with inhibitory concentrations in the range 0.001-1% (v/v). Among the single compounds, terpinen-4-ol showed the highest activity against Campylobacter spp. and other reference strains. Based on the antimicrobial activity and potential commerciality of these agents, lemon myrtle oil, α-tops (α-terpineol+cineole+terpinen-4-ol) and terpinen-4-ol were also evaluated using an in vitro fermentation technique to test antimicrobial activity towards C. jejuni in the microbiota from the chicken-caecum. EO compounds (terpinen-4-ol and α-tops) were antimicrobial towards C. jejuni at high doses (0.05%) without altering the fermentation profile. EOs and terpenoid compounds can have strong anti-Campylobacter activity without adversely affecting the fermentation potential of the chicken-caeca microbiota. EOs and their active compounds may have the potential to control C. jejuni colonisation and abundance in poultry.

  18. Antimicrobial activity of essential oils and five terpenoid compounds against Campylobacter jejuni in pure and mixed culture experiments.

    PubMed

    Kurekci, Cemil; Padmanabha, Jagadish; Bishop-Hurley, Sharon L; Hassan, Errol; Al Jassim, Rafat A M; McSweeney, Christopher S

    2013-09-16

    The aim of this study was to examine the antimicrobial potential of three essential oils (EOs: tea tree oil, lemon myrtle oil and Leptospermum oil), five terpenoid compounds (α-bisabolol, α-terpinene, cineole, nerolidol and terpinen-4-ol) and polyphenol against two strains of Campylobacter jejuni (ACM 3393 and the poultry isolate C338), Campylobacter coli and other Gram negative and Gram positive bacteria. Different formulations of neem oil (Azadirachta indica) with these compounds were also tested for synergistic interaction against all organisms. Antimicrobial activity was determined by the use of disc diffusion and broth dilution assays. All EOs tested were found to have strong antimicrobial activity against Campylobacter spp. with inhibitory concentrations in the range 0.001-1% (v/v). Among the single compounds, terpinen-4-ol showed the highest activity against Campylobacter spp. and other reference strains. Based on the antimicrobial activity and potential commerciality of these agents, lemon myrtle oil, α-tops (α-terpineol+cineole+terpinen-4-ol) and terpinen-4-ol were also evaluated using an in vitro fermentation technique to test antimicrobial activity towards C. jejuni in the microbiota from the chicken-caecum. EO compounds (terpinen-4-ol and α-tops) were antimicrobial towards C. jejuni at high doses (0.05%) without altering the fermentation profile. EOs and terpenoid compounds can have strong anti-Campylobacter activity without adversely affecting the fermentation potential of the chicken-caeca microbiota. EOs and their active compounds may have the potential to control C. jejuni colonisation and abundance in poultry. PMID:24041998

  19. Targeting motility properties of bacteria in the development of probiotic cultures against Campylobacter jejuni in broiler chickens.

    PubMed

    Aguiar, Vivian F; Donoghue, Ann M; Arsi, Komala; Reyes-Herrera, Ixchel; Metcalf, Joel H; de los Santos, Fausto S; Blore, Pamela J; Donoghue, Dan J

    2013-05-01

    Campylobacter is the leading cause of gastroenteritis worldwide. Campylobacter is commonly present in the intestinal tract of poultry, and one strategy to reduce enteric colonization is the use of probiotic cultures. This strategy has successfully reduced enteric colonization of Salmonella, but has had limited success against Campylobacter. In an effort to improve the efficacy of probiotic cultures, we developed a novel in vitro screening technique for selecting bacterial isolates with enhanced motility. It is proposed that motility-selected bacteria have the marked ability to reach the same gastrointestinal niche in poultry and competitively reduce C. jejuni. Bacterial isolates were collected from ceca of healthy chickens, and motile isolates were identified and tested for anti-Campylobacter activity. Isolates with these properties were selected for increased motility by passing each isolate 10 times and at each passage selecting bacteria that migrated the farthest during each passage. Three bacterial isolates with the greatest motility (all Bacillus subtilis) were used alone or in combination in two chicken trials. At day of hatch, chicks were administered these isolates alone or in combination (n=10/treatment, two trials), and chicks were orally challenged with a mixture of four different wild-type strains of C. jejuni (∼10(5) CFU/mL) on day 7. Isolate 1 reduced C. jejuni colonization in both of the trials (p<0.05). A follow-up study was conducted to compare isolate 1 subjected to enhanced motility selection with its nonselected form. A reduction (p<0.05) in Campylobacter colonization was observed in all three trials in the chickens dosed using isolate with enhanced motility compared to the control and unselected isolate. These findings support the theory that the motility enhancement of potential probiotic bacteria may provide a strategy for reduction of C. jejuni in preharvest chickens.

  20. Short-chain fatty acids and L-lactate as feed additives to control Campylobacter jejuni infections in broilers.

    PubMed

    Van Deun, Kim; Haesebrouck, Freddy; Van Immerseel, Filip; Ducatelle, Richard; Pasmans, Frank

    2008-08-01

    The usefulness of butyrate, acetate, propionate and l-lactate for the control of Campylobacter jejuni infections in broilers was assessed. For this purpose, the effect of these acids on the growth of C. jejuni in broth and intestinal mucous was determined, as well as their influence on the invasiveness of C. jejuni in intestinal epithelial cells. From these in vitro obtained results, one acid was retained for use as a feed additive in an in vivo trial. Butyrate was the most successful of the short-chain fatty acids, with 12.5 mM being bactericidal for C. jejuni at pH 6.0. Propionate and acetate had a bacteriostatic effect at 50 mM. None of the short-chain fatty acids had a bactericidal effect at pH 7.5 at a maximum concentration of 50 mM. Mucous increased the minimum bactericidal concentration of butyrate, but not the bacteriostatic concentrations of propionate or acetate. When C. jejuni was incubated in growth subinhibitory concentrations of butyrate, acetate or propionate or 25 mM L-lactate, no alteration in the invasive capabilities of C. jejuni in Caco-2 cells was observed. The addition of butyrate-coated micro-beads to the feed was unsuccessful to reduce C. jejuni caecal colonization in a seeder model using 2-week-old broilers. In conclusion, despite the marked bactericidal effect of butyrate towards C. jejuni in vitro, butyrate-coated micro-beads do not protect broilers from caecal colonization with C. jejuni in the applied test conditions. This might be partially ascribed to the protective effect of mucous and the rapid absorption of butyrate by the enterocytes. PMID:18622853

  1. Functional characterization of exopolyphosphatase/guanosine pentaphosphate phosphohydrolase (PPX/GPPA) of Campylobacter jejuni.

    PubMed

    Malde, Anandkumar; Gangaiah, Dharanesh; Chandrashekhar, Kshipra; Pina-Mimbela, Ruby; Torrelles, Jordi B; Rajashekara, Gireesh

    2014-05-15

    The inorganic polyphosphate (poly-P) is a key regulator of stress responses and virulence in many bacterial pathogens including Campylobacter jejuni. The role of exopolyphosphatases/guanosine pentaphosphate (pppGpp) phosphohydrolases (PPX/GPPA) in poly-P homeostasis and C. jejuni pathobiology remains unexplored. Here, we analyzed deletion mutants (∆ppx1, ∆ppx2) and the double knockout mutant (dkppx), all ∆ppx mutants exhibited increased capacity to accumulate poly-P; however only ∆ppx1 and dkppx mutants showed decreased accumulation of ppGpp, an alarmone molecule that regulates stringent response in bacteria, suggesting potential dual role for PPX1/GPPA. Nutrient survival defect of ∆ppx mutants was rescued by the supplementation of specific amino acids implying that survival defect may be associated with decreased ppGpp and/ or increased poly-P in ∆ppx mutants. The ppk1 and spoT were upregulated in both ∆ppx1 and ∆ppx2 suggesting a compensatory role for SpoT and Ppk1 in poly-P and ppGpp homeostasis. The lack of ppx genes resulted in defects in motility, biofilm formation, nutrient stress survival, invasion and intracellular survival indicating that maintaining a certain level of poly-P is critical for ppx genes in C. jejuni pathophysiology. Both ppx1 and ppx2 mutants were resistant to human complement-mediated killing; however, the dkppx mutant was sensitive. The serum susceptibility did not occur in the presence of MgCl 2 and EGTA suggesting an involvement of the classical or lectin pathway of complement mediated killing. Interestingly, the chicken serum did not have any effect on the ∆ppx mutants' survival. The observed serum susceptibility was not related to C. jejuni surface capsule and lipooligosaccharide structures. Our study underscores the importance of PPX/GPPA proteins in poly-P and ppGpp homeostasis, two critical molecules that modulate environmental stress responses and virulence in C. jejuni.

  2. Complete genome sequences of multidrug-resistant Campylobacter jejuni 14980A (turkey feces) and Campylobacter coli 14983A (housefly from turkey farm), harboring a novel gentamicin resistance mobile element.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Multidrug resistance (MDR) in foodborne pathogens is a major food safety and public health issue. Here we describe whole-genome sequences of two MDR strains of Campylobacter jejuni and Campylobacter coli from turkey feces and a housefly in a turkey farm. Both strains harbor a novel chromosomal genta...

  3. Complete Genome Sequences of Multidrug-Resistant Campylobacter jejuni Strain 14980A (Turkey Feces) and Campylobacter coli Strain 14983A (Housefly from a Turkey Farm), Harboring a Novel Gentamicin Resistance Mobile Element

    PubMed Central

    Huynh, Steven; Parker, Craig T.; Niedermeyer, Jeffrey A.; Kathariou, Sophia

    2016-01-01

    Multidrug resistance (MDR) in foodborne pathogens is a major food safety and public health issue. Here we describe whole-genome sequences of two MDR strains of Campylobacter jejuni and Campylobacter coli from turkey feces and a housefly from a turkey farm. Both strains harbor a novel chromosomal gentamicin resistance mobile element. PMID:27795285

  4. In vivo broiler experiments to assess anti-Campylobacter jejuni activity of a live Enterococcus faecalis strain.

    PubMed

    Robyn, J; Rasschaert, G; Hermans, D; Pasmans, F; Heyndrickx, M

    2013-01-01

    Bacterial gastroenteritis caused by thermotolerant Campylobacter species, mainly Campylobacter jejuni, has been the most reported zoonotic disease in many developed countries in recent years. Reducing Campylobacter shedding on the farm could result in a reduction of the number of campylobacteriosis cases. In 2 independent broiler seeder experiments, in which broiler chickens were orally inoculated with 2 amounts of Enterococcus faecalis MB 5259, we established whether a live E. faecalis strain was capable of reducing cecal Campylobacter colonization in broiler chickens. In previous in vitro experiments it has been demonstrated that this E. faecalis MB 5259 displays anti-Campylobacter activity. The effect of pH and bile salts on E. faecalis MB 5259 showed that growth and survival of E. faecalis MB 5259 can be impaired during passage through the gastrointestinal tract of broiler chickens. Despite these results E. faecalis MB 5259 was capable of colonizing the broiler ceca. Contrary to the in vitro experiments, in which E. faecalis MB 5259 inhibited C. jejuni MB 4185 growth, no inhibition was observed in the in vivo experiments independent of the inoculum size. PMID:23243257

  5. Detection of non-jejuni and -coli Campylobacter species from stool specimens with an immunochromatographic antigen detection assay.

    PubMed

    Couturier, Brianne A; Couturier, Marc Roger; Kalp, Kim J; Fisher, Mark A

    2013-06-01

    The STAT! Campy immunochromatographic assay for Campylobacter antigen was compared to culture for 500 clinical stool specimens. Antigen was detected in six culture-negative, PCR-positive specimens. C. upsaliensis, a pathogenic species that is traditionally difficult to recover in routine stool cultures, was detected in two of these culture-negative specimens. This study provides evidence that antigen testing may cross-react with at least one additional non-jejuni and -coli Campylobacter species that may be missed by routine culture for campylobacteriosis.

  6. Preliminary structural studies of the transcriptional regulator CmeR from Campylobacter jejuni

    SciTech Connect

    Su, Chih-Chia; Shi, Feng; Gu, Ruoyu; Li, Ming; McDermott, Gerry; Yu, Edward W.; Zhang, Qijing

    2007-01-01

    The transcriptional regulator CmeR from C. jejuni has been purified and crystallized and X-ray diffraction data have been collected to a resolution of 2.2 Å. In Campylobacter jejuni, a Gram-negative bacterial pathogen causing gastroenteritis in humans, the CmeR regulatory protein controls transcription of the multidrug transporter gene operon cmeABC. CmeR belongs to the TetR family of transcriptional regulators. The 210-residue CmeR consists of two functional motifs: an N-terminal DNA-binding domain and a C-terminal ligand-binding domain. It is predicted that the DNA-binding domain interacts directly with target promoters, while the C-terminal motif interacts with inducing ligands (such as bile salts). As an initial step towards confirming this structural model, recombinant CmeR protein containing a 6×His tag at the N-terminus was crystallized. Crystals of ligand-free CmeR belonged to space group P2{sub 1}2{sub 1}2, with unit-cell parameters a = 37.4, b = 57.6, c = 93.3 Å. Diffraction was observed to at least 2.2 Å at 100 K. Analysis of the detailed CmeR structure is currently in progress.

  7. High genetic similarity of ciprofloxacin-resistant Campylobacter jejuni in central Europe

    PubMed Central

    Kovač, Jasna; Čadež, Neža; Stessl, Beatrix; Stingl, Kerstin; Gruntar, Igor; Ocepek, Matjaž; Trkov, Marija; Wagner, Martin; Smole Možina, Sonja

    2015-01-01

    Campylobacteriosis is the leading zoonosis in the European Union with the majority of cases attributed to Campylobacter jejuni. Although the disease is usually self-limiting, some severe cases need to be treated with antibiotics, primarily macrolides and quinolones. However, the resistance to the latter is reaching alarming levels in most of the EU countries. To shed light on the expansion of antibiotic resistance in central Europe, we have investigated genetic similarity across 178 ciprofloxacin-resistant C. jejuni mostly isolated in Slovenia, Austria and Germany. We performed comparative genetic similarity analyses using allelic types of seven multilocus sequence typing housekeeping genes, and single nucleotide polymorphisms of a quinolone resistance determining region located within the DNA gyrase subunit A gene. This analysis revealed high genetic similarity of isolates from clonal complex ST-21 that carry gyrA allelic type 1 in all three of these central-European countries, suggesting these ciprofloxacin resistant isolates arose from a recent common ancestor and are spread clonally. PMID:26557112

  8. Emergence of variants with altered survival properties in stationary phase cultures of Campylobacter jejuni.

    PubMed

    Martínez-Rodriguez, A; Kelly, A F; Park, S F; Mackey, B M

    2004-02-01

    During the stationary phase of Campylobacter jejuni NCTC 11351 viable numbers fluctuate in a characteristic fashion. After reaching the maximum cell count (ca. 2 x 10(9) CFU/ml) in early stationary phase (denoted phase 1), viable numbers subsequently decrease to about 10(6) CFU/ml after 48 h and then increase again to about 10(8) CFU/ml (denoted phase 2) before decreasing once more to a value intermediate between the previous maximum and minimum values. To investigate whether the increase in viable numbers following the initial decline was due to the emergence of a new strain with a growth advantage in stationary phase analogous to the 'GASP' phenotype described in Escherichia coli [Science 259 (1993) 1757], we conducted mixed culture experiments with cells from the original culture and antibiotic-resistant marked organisms isolated from the re-growth phase. In many experiments of this type, strains isolated from phase 2 failed to out-compete the original strain and we have thus been unable to demonstrate a convincing GASP phenotype. However, strains isolated from phase 2 showed a much lower rate of viability loss in early stationary phase and a small increase in resistance to aeration, peroxide challenge and heat, indicating that the emergent strain was different from the parent. These results support the view that dynamic population changes occur during the stationary phase of C. jejuni that may play a role in the survival of this organism. PMID:14751687

  9. Multilocus Sequence Typing and Antimicrobial Resistance of Campylobacter jejuni Isolated from Dairy Calves in Austria

    PubMed Central

    Klein-Jöbstl, Daniela; Sofka, Dmitri; Iwersen, Michael; Drillich, Marc; Hilbert, Friederike

    2016-01-01

    Human campylobacteriosis is primarily associated with poultry but also cattle. In this study, 55 Campylobacter jejuni strains isolated from 382 dairy calves’ feces were differentiated by multilocus sequence typing and tested for antimicrobial resistance. The most prevalent sequence type (ST) was ST883 (20.0%), followed by ST48 (14.5%), and ST50 (9.1%). In contrast to ST48 and ST50, ST883 has rarely been described in cattle previously. Furthermore, risk factor analysis was performed for the presence of the most prevalent STs in these calves. Multiple regression analysis revealed that the type of farm (organic vs. conventional) and calf housing (place, and individual vs. group) were identified as significantly (p < 0.05) associated with the presence of ST883 in calves, and ST50 was associated with calf diarrhea. Antimicrobial resistance was detected in 58.2% of the isolates. Most of the resistant isolates (81.3%) were resistant to more than one antimicrobial. Most frequently, resistance to ciprofloxacin (49.1%), followed by nalidixic acid (42.8%), and tetracycline (14.5%) was observed. The results of the present study support the hypothesis that dairy calves may serve as a potential reservoir for C. jejuni and pose a risk for transmission, including antimicrobial resistant isolates to the environment and to humans. PMID:26870027

  10. Economical, simple method for production of the gaseous environment required for cultivation of Campylobacter jejuni.

    PubMed Central

    Pennie, R A; Zunino, J N; Rose, C E; Guerrant, R L

    1984-01-01

    Campylobacter jejuni is an enteric pathogen recognized worldwide as a cause of diarrhea. Its isolation from stool samples requires a microaerophilic environment that heretofore has been expensive and cumbersome to create. An economical, portable, and simple method is described which involves the production of appropriate concentrations of oxygen and carbon dioxide. Inside a plastic bag are placed two cups, one containing fine steel wool (grade 0) previously soaked in a 2.5% aqueous solution of copper sulfate and the other containing an Alka-Seltzer tablet in tap water. As suggested by Jurgensen et al. (Rev. Bras. Pat. Clin. 18:58-63, 1982), we used the effervescent antacid to generate CO2. By plate counts, we found this method to be as reliable in the cultivation of 20 isolates of C. jejuni in pure and mixed fecal culture as the reference gas method (85% N2, 10% CO2, and 5% O2). Analyses of the gas mixture inside the bag after up to 24 h of incubation confirmed the creation of an atmosphere of reduced O2 and increased CO2 concentrations. This method is eminently suitable for field situations in which more costly supplies are not available. PMID:6436301

  11. Economical, simple method for production of the gaseous environment required for cultivation of Campylobacter jejuni.

    PubMed

    Pennie, R A; Zunino, J N; Rose, C E; Guerrant, R L

    1984-09-01

    Campylobacter jejuni is an enteric pathogen recognized worldwide as a cause of diarrhea. Its isolation from stool samples requires a microaerophilic environment that heretofore has been expensive and cumbersome to create. An economical, portable, and simple method is described which involves the production of appropriate concentrations of oxygen and carbon dioxide. Inside a plastic bag are placed two cups, one containing fine steel wool (grade 0) previously soaked in a 2.5% aqueous solution of copper sulfate and the other containing an Alka-Seltzer tablet in tap water. As suggested by Jurgensen et al. (Rev. Bras. Pat. Clin. 18:58-63, 1982), we used the effervescent antacid to generate CO2. By plate counts, we found this method to be as reliable in the cultivation of 20 isolates of C. jejuni in pure and mixed fecal culture as the reference gas method (85% N2, 10% CO2, and 5% O2). Analyses of the gas mixture inside the bag after up to 24 h of incubation confirmed the creation of an atmosphere of reduced O2 and increased CO2 concentrations. This method is eminently suitable for field situations in which more costly supplies are not available. PMID:6436301

  12. Investigations into the antibacterial activities of phytotherapeutics against Helicobacter pylori and Campylobacter jejuni.

    PubMed

    Cwikla, C; Schmidt, K; Matthias, A; Bone, K M; Lehmann, R; Tiralongo, E

    2010-05-01

    The prevalence of gastric diseases is increasing with H. pylori, the causative agent of acute and chronic gastritis, being a major predisposing factor for peptic ulcer disease and gastric carcinoma. C. jejuni is the most common cause of enteric infections, particularly among children, resulting in severe diarrhoea. Increasing drug resistance of these bacteria against standard antibiotics, and the more widespread use of herbal medicines, favours investigations into additional anti-Helicobacter and anti-Campylobacter effects of phytotherapeutics that are already used for their beneficial effects on bowel and digestive functions. Twenty-one hydroethanol herbal extracts and four essential oils were screened for antibacterial activity using a modification of a previously described micro-dilution assay and compared with the inhibitory effects of antibiotics. The herbal extracts showing the highest growth inhibition of C. jejuni were Calendula officinalis, Matricaria recutita, Zingiber officinale, Salvia officinalis, Foeniculum vulgare and Silybum marianum. Agrimonia eupatoria, Hydrastis canadensis, Filipendula ulmaria and Salvia officinalis were the most active herbal extracts in inhibiting the growth of H. pylori. This study provides evidence for additional beneficial effects of phytotherapeutics marketed for their gastrointestinal effects and identifies new beneficial antibacterial effects for some herbal medicines not currently recommended for gastrointestinal problems. PMID:19653313

  13. The impairment of methylmenaquinol:fumarate reductase affects hydrogen peroxide susceptibility and accumulation in Campylobacter jejuni.

    PubMed

    Kassem, Issmat I; Khatri, Mahesh; Sanad, Yasser M; Wolboldt, Melinda; Saif, Yehia M; Olson, Jonathan W; Rajashekara, Gireesh

    2014-04-01

    The methylmenaquinol:fumarate reductase (Mfr) of Campylobacter jejuni is a periplasmic respiratory (redox) protein that contributes to the metabolism of fumarate and displays homology to succinate dehydrogenase (Sdh). Since chemically oxidized redox-enzymes, including fumarate reductase and Sdh, contribute to the generation of oxidative stress in Escherichia coli, we assessed the role of Mfr in C. jejuni after exposure to hydrogen peroxide (H2 O2 ). Our results show that a Mfr mutant (∆mfrA) strain was less susceptible to H2 O2 as compared to the wildtype (WT). Furthermore, the H2 O2 concentration in the ∆mfrA cultures was significantly higher than that of WT after exposure to the oxidant. In the presence of H2 O2 , catalase (KatA) activity and katA expression were significantly lower in the ∆mfrA strain as compared to the WT. Exposure to H2 O2 resulted in a significant decrease in total intracellular iron in the ∆mfrA strain as compared to WT, while the addition of iron to the growth medium mitigated H2 O2 susceptibility and accumulation in the mutant. The ∆mfrA strain was significantly more persistent in RAW macrophages as compared to the WT. Scanning electron microscopy showed that infection with the ∆mfrA strain caused prolonged changes to the macrophages' morphology, mainly resulting in spherical-shaped cells replete with budding structures and craters. Collectively, our results suggest a role for Mfr in maintaining iron homeostasis in H2 O2 stressed C. jejuni, probably via affecting the concentrations of intracellular iron. PMID:24515965

  14. Lipooligosaccharide locus class of Campylobacter jejuni: sialylation is not needed for invasive infection.

    PubMed

    Ellström, P; Feodoroff, B; Hänninen, M-L; Rautelin, H

    2014-06-01

    Campylobacter jejuni is a highly diverse enteropathogen that is commonly detected worldwide. It can sometimes cause bacteraemia, but the bacterial characteristics facilitating bloodstream infection are not known. A total of 73 C. jejuni isolates, consecutively collected from blood-borne infections during a 10-year period all over Finland and for which detailed clinical information of the patients were available, were included. We screened the isolates by PCR for the lipooligosaccharide (LOS) locus class and for the presence of the putative virulence genes ceuE, ciaB, fucP, and virB11. The isolates were also tested for γ-glutamyl transpeptidase production. The results were analysed with respect to the clinical characteristics of the patients, and the multilocus sequence types (MLSTs) and serum resistance of the isolates. LOS locus classes A, B, and C, which carry genes for sialylation of LOS, were detected in only 23% of the isolates. These isolates were not more resistant to human serum than those with the genes of non-sialylated LOS locus classes, but were significantly more prevalent among patients with underlying diseases (p 0.02). The fucose permease gene fucP was quite uncommon, but was associated with the isolates with the potential to sialylate LOS (p <0.0001). LOS locus classes and some of the putative virulence factors were associated with MLST clonal complexes. Although some of the bacterial characteristics studied here have been suggested to be important for the invasiveness of C. jejuni, they did not explain why the clinical isolates in the present study were able to cause bacteraemia.

  15. Specificity of Campylobacter jejuni Adhesin PEB3 for Phosphates and Structural Differences among Its Ligand Complexes

    SciTech Connect

    Min, Tongpil; Vedadi, Masoud; Watson, David C.; Wasney, Gregory A.; Munger, Christine; Cygler, Miroslaw; Matte, Allan; Young, N. Martin

    2009-04-22

    PEB3 is a glycoprotein adhesin from Campylobacter jejuni whose structure suggested a role in transport. We have investigated potential ligands for PEB3 and characterized their binding properties using biophysical methods in solution and by X-ray crystallography. A thermal aggregation assay of PEB3 with a library of physiological compounds identified three possible ligands [3-phosphoglycerate (3-PG), phosphoenolpyruvate (PEP), and aconitate], which stabilized wild-type PEB3 but did not stabilize either a PEB3 form containing two mutations at the ligand-binding site, T138A/S139A, or a second PEB3 mutant, K135E, at a site {approx}14 {angstrom} away. Fluorescence titration experiments and cocrystal structures with various ligands were used to characterize the binding of 3-PG, PEP, and phosphate to PEB3. Further, a C. jejuni growth experiment in minimal medium supplemented with 3-PG showed that this molecule enhances the growth of wild-type C. jejuni, but not of the PEB3 mutants. Crystallographic analysis of PEB3 complexes revealed that the Ser171-Gln180 region in the presence of 3-PG or other phosphates is helical and similar to those of other transport proteins, but it is nonhelical when citrate is bound. The K135E mutation resulted in expression of a more highly glycosylated form of PEB3 in vivo, and its crystal structure showed the conformation of the first two residues of the glycan. On the basis of our findings, we suggest that PEB3 is a transport protein that may function in utilization of 3-PG or other phosphate-containing molecules from the host.

  16. Molecular epidemiology of Campylobacter jejuni in a geographically isolated country with a uniquely structured poultry industry.

    PubMed

    Müllner, Petra; Collins-Emerson, Julie M; Midwinter, Anne C; Carter, Philip; Spencer, Simon E F; van der Logt, Peter; Hathaway, Steve; French, Nigel P

    2010-04-01

    In New Zealand the number of campylobacteriosis notifications increased markedly between 2000 and 2007. Notably, this country's poultry supply is different than that of many developed countries as the fresh and frozen poultry available at retail are exclusively of domestic origin. To examine the possible link between human cases and poultry, a sentinel surveillance site was established to study the molecular epidemiology of Campylobacter jejuni over a 3-year period from 2005 to 2008 using multilocus sequence typing. Studies showed that 60.1 to 81.4% of retail poultry carcasses from the major suppliers were contaminated with C. jejuni. Differences were detected in the probability and level of contamination and the relative frequency of genotypes for individual poultry suppliers and humans. Some carcasses were contaminated with isolates belonging to more than one sequence type (ST), and there was evidence of both ubiquitous and supplier-associated strains, an epidemiological pattern not recognized yet in other countries. The common poultry STs were also common in human clinical cases, providing evidence that poultry is a major contributor to human infection. Both internationally rare genotypes, such as ST-3069 and ST-474, and common genotypes, such as ST-45 and ST-48, were identified in this study. The dominant human sequence type in New Zealand, ST-474, was found almost exclusively in isolates from one poultry supplier, which provided evidence that C. jejuni has a distinctive molecular epidemiology in this country. These results may be due in part to New Zealand's geographical isolation and its uniquely structured poultry industry. PMID:20154115

  17. High-Frequency Variation of Purine Biosynthesis Genes Is a Mechanism of Success in Campylobacter jejuni

    PubMed Central

    Cameron, Andrew; Huynh, Steven; Scott, Nichollas E.; Frirdich, Emilisa; Apel, Dmitry; Foster, Leonard J.; Parker, Craig T.

    2015-01-01

    ABSTRACT Phenotypic variation is prevalent in the zoonotic pathogen Campylobacter jejuni, the leading agent of enterocolitis in the developed world. Heterogeneity enhances the survival and adaptive malleability of bacterial populations because variable phenotypes may allow some cells to be protected against future stress. Exposure to hyperosmotic stress previously revealed prevalent differences in growth between C. jejuni strain 81-176 colonies due to resistant or sensitive phenotypes, and these isolated colonies continued to produce progeny with differential phenotypes. In this study, whole-genome sequencing of isolated colonies identified allelic variants of two purine biosynthesis genes, purF and apt, encoding phosphoribosyltransferases that utilize a shared substrate. Genetic analyses determined that purF was essential for fitness, while apt was critical. Traditional and high-depth amplicon-sequencing analyses confirmed extensive intrapopulation genetic variation of purF and apt that resulted in viable strains bearing alleles with in-frame insertion duplications, deletions, or missense polymorphisms. Different purF and apt alleles were associated with various stress survival capabilities under several niche-relevant conditions and contributed to differential intracellular survival in an epithelial cell infection model. Amplicon sequencing revealed that intracellular survival selected for stress-fit purF and apt alleles, as did exposure to oxygen and hyperosmotic stress. Putative protein recognition direct repeat sequences were identified in purF and apt, and a DNA-protein affinity screen captured a predicted exonuclease that promoted the global spontaneous mutation rate. This work illustrates the adaptive properties of high-frequency genetic variation in two housekeeping genes, which influences C. jejuni survival under stress and promotes its success as a pathogen. PMID:26419875

  18. Campylobacter jejuni pdxA Affects Flagellum-Mediated Motility to Alter Host Colonization

    PubMed Central

    Asakura, Hiroshi; Hashii, Noritaka; Uema, Masashi; Kawasaki, Nana; Sugita-Konishi, Yoshiko; Igimi, Shizunobu; Yamamoto, Shigeki

    2013-01-01

    Vitamin B6 (pyridoxal-5'-phosphate, PLP) is linked to a variety of biological functions in prokaryotes. Here, we report that the pdxA (putative 4-hydroxy-L-threonine phosphate dehydrogenase) gene plays a pivotal role in the PLP-dependent regulation of flagellar motility, thereby altering host colonization in a leading foodborne pathogen, Campylobacter jejuni. A C. jejuni pdxA mutant failed to produce PLP and exhibited a coincident loss of flagellar motility. Mass spectrometric analyses showed a 3-fold reduction in the main flagellar glycan pseudaminic acid (Pse) associated with the disruption of pdxA. The pdxA mutant also exhibited reduced growth rates compared with the WT strain. Comparative metabolomic analyses revealed differences in respiratory/energy metabolism between WT C. jejuni and the pdxA mutant, providing a possible explanation for the differential growth fitness between the two strains. Consistent with the lack of flagellar motility, the pdxA mutant showed impaired motility-mediated responses (bacterial adhesion, ERK1/2 activation, and IL-8 production) in INT407 cells and reduced colonization of chickens compared with the WT strain. Overall, this study demonstrated that the pdxA gene affects the PLP-mediated flagellar motility function, mainly through alteration of Pse modification, and the disruption of this gene also alters the respiratory/energy metabolisms to potentially affect host colonization. Our data therefore present novel implications regarding the utility of PLP and its dependent enzymes as potent target(s) for the control of this pathogen in the poultry host. PMID:23936426

  19. A PCR-RFLP assay for the detection and differentiation of Campylobacter jejuni, C. coli, C. fetus, C. hyointestinalis, C. lari, C. helveticus and C. upsaliensis.

    PubMed

    Kamei, Kazumasa; Asakura, Masahiro; Somroop, Srinuan; Hatanaka, Noritoshi; Hinenoya, Atsushi; Nagita, Akira; Misawa, Naoaki; Matsuda, Motoo; Nakagawa, Shinsaku; Yamasaki, Shinji

    2014-05-01

    Although Campylobacter jejuni and Campylobacter coli are the most common bacterial causes of human gastrointestinal diseases, other Campylobacter species are also involved in human and animal infections. In this study, we developed a cytolethal distending toxin (cdt) gene-based PCR-RFLP assay for the detection and differentiation of C. jejuni, C. coli, C. fetus, C. hyointestinalis, C. lari, C. helveticus and C. upsaliensis. Previously designed common primers, which can amplify the cdtB gene of C. jejuni, C. coli and C. fetus, were used for detecting seven Campylobacter species and differentiating between them by restriction digestion. The PCR-RFLP assay was validated with 277 strains, including 35 C. jejuni, 19 C. coli, 20 C. fetus, 24 C. hyointestinalis, 13 C. lari, 2 C. helveticus, 22 C. upsaliensis, 3 other Campylobacter spp. and 17 other species associated with human diseases. Sensitivity and specificity of the PCR-RFLP assay were 100 % except for C. hyointestinalis (88 % sensitivity). Furthermore, the PCR-RFLP assay successfully detected and differentiated C. jejuni, C. coli and C. fetus in clinical and animal samples. The results indicate that the PCR-RFLP assay is useful for the detection and differentiation of seven Campylobacter species important for human and animal diseases.

  20. A SpoT polymorphism correlates with chill stress survival and is prevalent in clinical isolates of Campylobacter jejuni.

    PubMed

    Nierop Groot, M N; de Boer, A G; van Pelt, W; van der Hulst-van Arkel, M C; de Leeuw, P; Widjaja, H C A; Smits, M A; van der Wal, F J

    2014-11-01

    Resistance of Campylobacter jejuni to environmental stress is regarded as a risk factor for the transmission of C. jejuni from poultry or poultry products to humans. So far, the mechanisms underlying the capacity of C. jejuni to survive environmental stress conditions are not fully understood. In this study, we searched for polymorphisms in C. jejuni genes, potentially involved in resistance to chill stress. To this end, we assessed 3 groups of C. jejuni isolates (clinical, retail chicken meat, and feces) for survival of experimentally induced chill stress. For each isolate we sequenced 3 genes encoding the C. jejuni sigma factors FliA, RpoD, and RpoN as well as the genes for the transcriptional regulator SpoT and the periplasmic protein HtrA. Data suggest a higher prevalence of a specific polymorphism in spoT in clinical isolates compared with poultry meat or farm isolates. Moreover, this genotype correlated with enhanced survival of chill stress. The observation that the prevalence of this SNP is relatively high in clinical isolates, which most likely have been exposed to multiple forms of stress, suggest that this SNP may be a biomarker for enhanced survival of stress.

  1. [Use of AFLP analysis for the molecular biological fine typing of Campylobacter jejuni strains from turkey flocks].

    PubMed

    Alter, Thomas; Fehlhaber, Karsten

    2003-01-01

    One of the most promising genotyping methods for microorganisms is the AFLP (amplified fragment length polymorphism)-analysis, originally developed to genotype plants. The AFLP method has a high discriminatory power and an excellent reproducibility. Thus it is suitable for solving taxonomic and epidemiological questions. AFLP analysis was used to reveal the genetic diversity of Campylobacter (C.) jejuni strains from turkeys. The aim was to detect and genotype C. jejuni strains on individual turkeys throughout the complete slaughter process. A broad spectrum of genotypes was detectable on the turkey skin at the beginning of the slaughter line. At the end of the slaughter process-after chilling, only a diminished spectrum of genotypes was detectable by AFLP-analysis, suggesting, that slaughter specific environmental stressors may exert a selective pressure on the strain diversity. Our results proved, that AFLP analysis is an excellent tool for subtyping large numbers of Campylobacter strains. PMID:14655622

  2. Multiplex strategy for multilocus sequence typing, fla typing, and genetic determination of antimicrobial resistance of Campylobacter jejuni and Campylobacter coli isolates collected in Switzerland.

    PubMed

    Korczak, Bozena M; Zurfluh, Monika; Emler, Stefan; Kuhn-Oertli, Jacqueline; Kuhnert, Peter

    2009-07-01

    We present an optimized multilocus sequence typing (MLST) scheme with universal primer sets for amplifying and sequencing the seven target genes of Campylobacter jejuni and Campylobacter coli. Typing was expanded by sequence determination of the genes flaA and flaB using optimized primer sets. This approach is compatible with the MLST and flaA schemes used in the PubMLST database and results in an additional typing method using the flaB gene sequence. An identification module based on the 16S rRNA and rpoB genes was included, as well as the genetic determination of macrolide and quinolone resistances based on mutations in the 23S rRNA and gyrA genes. Experimental procedures were simplified by multiplex PCR of the 13 target genes. This comprehensive approach was evaluated with C. jejuni and C. coli isolates collected in Switzerland. MLST of 329 strains resulted in 72 sequence types (STs) among the 186 C. jejuni strains and 39 STs for the 143 C. coli isolates. Fourteen (19%) of the C. jejuni and 20 (51%) of the C. coli STs had not been found previously. In total, 35% of the C. coli strains collected in Switzerland contained mutations conferring antibiotic resistance only to quinolone, 15% contained mutations conferring resistance only to macrolides, and 6% contained mutations conferring resistance to both classes of antibiotics. In C. jejuni, these values were 31% and 0% for quinolone and macrolide resistance, respectively. The rpoB sequence allowed phylogenetic differentiation between C. coli and C. jejuni, which was not possible by 16S rRNA gene analysis. An online Integrated Database Network System (SmartGene, Zug, Switzerland)-based platform for MLST data analysis specific to Campylobacter was implemented. This Web-based platform allowed automated allele and ST designation, as well as epidemiological analysis of data, thus streamlining and facilitating the analysis workflow. Data networking facilitates the exchange of information between collaborating centers

  3. Epidemiological investigations on Campylobacter jejuni in households with a primary infection.

    PubMed Central

    Oosterom, J.; den Uyl, C. H.; Bänffer, J. R.; Huisman, J.

    1984-01-01

    Fifty-four Rotterdam patients in which a primary infection with Campylobacter jejuni had been detected (index patients) were compared with 54 control subjects with regard to the consumption and preparation of foods 7 days before onset of illness and the keeping of pet animals. Significantly more index patients than controls had eaten chicken meat (47 v. 29; P = 0.0002), particularly at barbecues (14 v. 2; P = 0.0015). Marginally more index patients had eaten pork (47 v. 39; P = 0.048) or inadequately heated meat (13 v. 8), though in the last case numbers were too small to be statistically significant. The consumption of beef or mutton and outdoor eating (other than at barbecues) were essentially the same in both groups. There was no significant association with the keeping of pet animals, although a few more index patients had cage birds than controls (18 v. 12). Twenty-one (15%) of 136 household contacts of index patients also suffered from diarrhoea during the same period. Circumstantial evidence pointed to a common source of infection with the index patient in 13 instances (nine households) and probable intrafamilial spread of infection in six instances. Campylobacters were isolated from one of 110 swabs of kitchen work surfaces and eight of 107 swabs taken from lavatory bowls in index households. PMID:6501878

  4. The evidence for clonal spreading of quinolone resistance with a particular clonal complex of Campylobacter jejuni.

    PubMed

    Kovač, J; Cadež, N; Lušicky, M; Nielsen, E Møller; Ocepek, M; Raspor, P; Možina, S Smole

    2014-12-01

    Campylobacter is the most prevalent cause of bacterial gastroenteritis worldwide and it represents a significant public health risk of increasing severity due to its escalating resistance to clinically important quinolone and macrolide antibiotics. As a zoonotic pathogen Campylobacter is transmitted along the food chain and naturally cycles from environmental waters, feedstuff, animals and food to humans. We determined antibiotic resistance profiles, as well as multilocus sequence types and flaA-SVR types for 52 C. jejuni isolated in Slovenia from human, animal, raw and cured chicken meat and water samples. Twenty-eight different sequence types, arranged in ten clonal complexes, three new allele types and five new sequence types were identified, indicating the relatively high diversity in a small group of strains. The assignment of strains from different sources to the same clonal complexes indicates their transmission along the food supply chain. The most prevalent clonal complex was CC21, which was also the genetic group with 95% of quinolone-resistant strains. Based on the genetic relatedness of these quinolone-resistant strains identified by polymerase chain reaction with a mismatch amplification mutation assay and sequencing of the quinolone resistance-determining region of the gyrA gene, we conclude that the high resistance prevalence observed indicates the local clonal spread of quinolone resistance with CC21. PMID:24534165

  5. Enhanced biofilm formation and multi-host transmission evolve from divergent genetic backgrounds in Campylobacter jejuni.

    PubMed

    Pascoe, Ben; Méric, Guillaume; Murray, Susan; Yahara, Koji; Mageiros, Leonardos; Bowen, Ryan; Jones, Nathan H; Jeeves, Rose E; Lappin-Scott, Hilary M; Asakura, Hiroshi; Sheppard, Samuel K

    2015-11-01

    Multicellular biofilms are an ancient bacterial adaptation that offers a protective environment for survival in hostile habitats. In microaerophilic organisms such as Campylobacter, biofilms play a key role in transmission to humans as the bacteria are exposed to atmospheric oxygen concentrations when leaving the reservoir host gut. Genetic determinants of biofilm formation differ between species, but little is known about how strains of the same species achieve the biofilm phenotype with different genetic backgrounds. Our approach combines genome-wide association studies with traditional microbiology techniques to investigate the genetic basis of biofilm formation in 102 Campylobacter jejuni isolates. We quantified biofilm formation among the isolates and identified hotspots of genetic variation in homologous sequences that correspond to variation in biofilm phenotypes. Thirteen genes demonstrated a statistically robust association including those involved in adhesion, motility, glycosylation, capsule production and oxidative stress. The genes associated with biofilm formation were different in the host generalist ST-21 and ST-45 clonal complexes, which are frequently isolated from multiple host species and clinical samples. This suggests the evolution of enhanced biofilm from different genetic backgrounds and a possible role in colonization of multiple hosts and transmission to humans.

  6. Membrane integrity of Campylobacter jejuni subjected to high pressure is pH-dependent

    NASA Astrophysics Data System (ADS)

    Lerasle, M.; Guillou, S.; Simonin, H.; Laroche, M.; de Lamballerie, M.; Federighi, M.

    2012-03-01

    Our study focuses on a foodborne pathogen, Campylobacter, which is responsible for the most frequent bacterial enteritis worldwide. Membrane integrity of Campylobacter jejuni NCTC 11168 cells treated at high pressure (300 MPa, 20°C, 10 min) at pH 7.0 and pH 5.6 was measured by fluorescence spectroscopy of propidium iodide (PI) uptake. The percentage of membrane-damaged cells by high pressure, in which PI is allowed to penetrate, was determined using two calibration methods based on the PI fluorescence signal obtained with cells killed either by a heat treatment (80°C for 15 min) or by a pressure treatment (400 MPa, 20°C, 10 min). Both calibrations were shown to be statistically different (P<0.05), particularly at acidic pH, suggesting that a difference in the penetration of PI into bacterial cells might depend on the mode of cell inactivation. These results corroborate the fact that the mechanism of microbial inactivation by high pressure is pH-dependent.

  7. Campylobacter jejuni colonization promotes the translocation of Escherichia coli to extra-intestinal organs and disturbs the short-chain fatty acids profiles in the chicken gut.

    PubMed

    Awad, W A; Dublecz, F; Hess, C; Dublecz, K; Khayal, B; Aschenbach, J R; Hess, M

    2016-10-01

    For a long time Campylobacter was only considered as a commensal microorganism in avian hosts restricted to the ceca, without any pathogenic features. The precise reasons for the symptomless chicken carriers are still unknown, but investigations of the gastrointestinal ecology of broiler chickens may improve our understanding of the microbial interactions with the host. Therefore, the current studies were conducted to investigate the effects of Campylobacter jejuni colonization on Escherichia coli translocation and on the metabolic end products (short-chain fatty acids, SCFAs). Following oral infection of 14 day old broiler chickens with 1 × 10(8) CFU of Campylobacter jejuni NCTC 12744 in two independent animal trials, it was found that C. jejuni heavily colonized the intestine and disseminate to extra-intestinal organs. Moreover, in both animal trials, the findings revealed that C. jejuni promoted the translocation of E. coli with a higher number encountered in the spleen and liver at 14 days post infection (dpi). In addition, Campylobacter affected the microbial fermentation in the gastrointestinal tract of broilers by reducing the amount of propionate, isovalerate, and isobutyrate in the cecal digesta of the infected birds at 2 dpi and, at 7 and 14 dpi, butyrate, isobutyrate, and isovalerate were also decreased. However, in the jejunum, the C. jejuni infection lowered only butyrate concentrations at 14 dpi. These data indicated that C. jejuni may utilize SCFAs as carbon sources to promote its colonization in the chicken gut, suggesting that Campylobacter cannot only alter gut colonization dynamics but might also influence physiological processes due to altered microbial metabolite profiles.Finally, the results demonstrated that C. jejuni can cross the intestinal epithelial barrier and facilitates the translocation of Campylobacter itself as well as of other enteric microorganisms such as E. coli to extra-intestinal organs of infected birds. Altogether, our

  8. Genetic Diversity and Antimicrobial Susceptibility of Campylobacter jejuni Isolates Associated with Sheep Abortion in the United States and Great Britain

    PubMed Central

    Wu, Zuowei; Sippy, Rachel; Plummer, Paul; Vidal, Ana; Newell, Diane; Zhang, Qijing

    2014-01-01

    Campylobacter infection is a leading cause of ovine abortion worldwide. Historically, genetically diverse Campylobacter fetus and Campylobacter jejuni strains have been implicated in such infections, but since 2003 a highly pathogenic, tetracycline-resistant C. jejuni clone (named SA) has become the predominant cause of sheep abortions in the United States. Whether clone SA was present in earlier U.S. abortion isolates (before 2000) and is associated with sheep abortions outside the United States are unknown. Here, we analyzed 54 C. jejuni isolates collected from U.S. sheep abortions at different time periods and compared them with 42 C. jejuni isolates associated with sheep abortion during 2002 to 2008 in Great Britain, using multilocus sequence typing (MLST), pulsed-field gel electrophoresis (PFGE), and array-based comparative genomic hybridization (CGH). Although clone SA (ST-8) was present in the early U.S. isolates, it was not as tetracycline resistant (19% versus 100%) or predominant (66% versus 91%) as it was in the late U.S isolates. In contrast, C. jejuni isolates from Great Britain were genetically diverse, comprising 19 STs and lacking ST-8. PFGE and CGH analyses of representative strains further confirmed the population structure of the abortion isolates. Notably, the Great Britain isolates were essentially susceptible to most tested antibiotics, including tetracycline, while the late U.S. isolates were universally resistant to this antibiotic, which could be explained by the common use of tetracyclines for control of sheep abortions in the United States but not in Great Britain. These results suggest that the dominance of clone SA in sheep abortions is unique to the United States, and the use of tetracyclines may have facilitated selection of this highly pathogenic clone. PMID:24648552

  9. Genetic diversity and antimicrobial susceptibility of Campylobacter jejuni isolates associated with sheep abortion in the United States and Great Britain.

    PubMed

    Wu, Zuowei; Sippy, Rachel; Sahin, Orhan; Plummer, Paul; Vidal, Ana; Newell, Diane; Zhang, Qijing

    2014-06-01

    Campylobacter infection is a leading cause of ovine abortion worldwide. Historically, genetically diverse Campylobacter fetus and Campylobacter jejuni strains have been implicated in such infections, but since 2003 a highly pathogenic, tetracycline-resistant C. jejuni clone (named SA) has become the predominant cause of sheep abortions in the United States. Whether clone SA was present in earlier U.S. abortion isolates (before 2000) and is associated with sheep abortions outside the United States are unknown. Here, we analyzed 54 C. jejuni isolates collected from U.S. sheep abortions at different time periods and compared them with 42 C. jejuni isolates associated with sheep abortion during 2002 to 2008 in Great Britain, using multilocus sequence typing (MLST), pulsed-field gel electrophoresis (PFGE), and array-based comparative genomic hybridization (CGH). Although clone SA (ST-8) was present in the early U.S. isolates, it was not as tetracycline resistant (19% versus 100%) or predominant (66% versus 91%) as it was in the late U.S isolates. In contrast, C. jejuni isolates from Great Britain were genetically diverse, comprising 19 STs and lacking ST-8. PFGE and CGH analyses of representative strains further confirmed the population structure of the abortion isolates. Notably, the Great Britain isolates were essentially susceptible to most tested antibiotics, including tetracycline, while the late U.S. isolates were universally resistant to this antibiotic, which could be explained by the common use of tetracyclines for control of sheep abortions in the United States but not in Great Britain. These results suggest that the dominance of clone SA in sheep abortions is unique to the United States, and the use of tetracyclines may have facilitated selection of this highly pathogenic clone.

  10. Designing multiplex PCR system of Campylobacter jejuni for efficient typing by improving monoplex PCR binary typing method.

    PubMed

    Yamada, Kazuhiro; Ibata, Ami; Suzuki, Masahiro; Matsumoto, Masakado; Yamashita, Teruo; Minagawa, Hiroko; Kurane, Ryuichiro

    2015-01-01

    Campylobacter jejuni is responsible for the majority of Campylobacter infections. As the molecular epidemiological study of outbreaks, pulsed-field gel electrophoresis (PFGE) is performed in general. But PFGE has several problems. PCR binary typing (P-BIT) method is a typing method for Campylobacter spp. that was recently developed, and was reported to have a similar discriminatory power and stability to those of PFGE. We modified the P-BIT method from 18 monoplex PCRs to two multiplex PCR systems (mP-BIT). The same results were obtained from monoplex PCRs using original primers and multiplex PCR in the representative isolates. The mP-BIT can analyze 48 strains at a time by using 96-well PCR systems and can identify C. jejuni because mP-BIT includes C. jejuni marker. The typing of the isolates by the mP-BIT and PFGE demonstrated generally concordant results and the mP-BIT method (D = 0.980) has a similar discriminatory power to that of PFGE with SmaI digest (D = 0.975) or KpnI digest (D = 0.987) as with original article. The mP-BIT method is quick, simple and easy, and comes to be able to perform it at low cost by having become a multiplex PCR system. Therefore, the mP-BIT method with two multiplex PCR systems has high potential for a rapid first-line surveillance typing assay of C. jejuni and can be used for routine surveillance and outbreak investigations of C. jejuni in the future.

  11. 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.

  12. Designing multiplex PCR system of Campylobacter jejuni for efficient typing by improving monoplex PCR binary typing method.

    PubMed

    Yamada, Kazuhiro; Ibata, Ami; Suzuki, Masahiro; Matsumoto, Masakado; Yamashita, Teruo; Minagawa, Hiroko; Kurane, Ryuichiro

    2015-01-01

    Campylobacter jejuni is responsible for the majority of Campylobacter infections. As the molecular epidemiological study of outbreaks, pulsed-field gel electrophoresis (PFGE) is performed in general. But PFGE has several problems. PCR binary typing (P-BIT) method is a typing method for Campylobacter spp. that was recently developed, and was reported to have a similar discriminatory power and stability to those of PFGE. We modified the P-BIT method from 18 monoplex PCRs to two multiplex PCR systems (mP-BIT). The same results were obtained from monoplex PCRs using original primers and multiplex PCR in the representative isolates. The mP-BIT can analyze 48 strains at a time by using 96-well PCR systems and can identify C. jejuni because mP-BIT includes C. jejuni marker. The typing of the isolates by the mP-BIT and PFGE demonstrated generally concordant results and the mP-BIT method (D = 0.980) has a similar discriminatory power to that of PFGE with SmaI digest (D = 0.975) or KpnI digest (D = 0.987) as with original article. The mP-BIT method is quick, simple and easy, and comes to be able to perform it at low cost by having become a multiplex PCR system. Therefore, the mP-BIT method with two multiplex PCR systems has high potential for a rapid first-line surveillance typing assay of C. jejuni and can be used for routine surveillance and outbreak investigations of C. jejuni in the future. PMID:25455748

  13. Experimental Campylobacter jejuni infection in the chicken: an animal model of axonal Guillain-Barré syndrome.

    PubMed Central

    Li, C Y; Xue, P; Tian, W Q; Liu, R C; Yang, C

    1996-01-01

    OBJECTIVE: To develop and characterise an animal model of paralytic neuropathy after Campylobacter jejuni infection. Campylobacter infection precedes development of many cases of Guillain-Barré syndrome and is particularly associated with cases having prominent axonal degeneration. Understanding the pathogenesis of Guillain-Barré syndrome after C jejuni infection has been slowed by the lack of animal models. METHODS: A spontaneous paralytic neuropathy is described that developed in chickens from the farms of four patients with Guillain-Barré syndrome. The production of paralytic neuropathy in chickens experimentally fed Campylobacter jejuni isolated from one of these patients is reported. The sciatic nerves of the spontaneously paralysed chickens were examined pathologically in teased fibres, in plastic embedded sections, and by electron microscopy. Two large groups of chickens were then fed cultures of a C jejuni (Penner type O:19) isolated from one of these patients. RESULTS: The chickens with spontaneous paralysis had pathologically noninflammatory neuropathy. Pathology in the sciatic nerves ranged from no detectable changes to severe Wallerian-like degeneration. In the experimentally inoculated groups, an average of 33% of the chickens became paralysed. The median time after inoculation to paralysis was 12 days. The lesions found in the first few days of paralysis included nodal lengthening and paranodal demyelination. In those animals that survived for several days after onset of weakness, the pathology was dominated by extensive Wallerian-like degeneration. Animals that survived for weeks with no clinically apparent neuropathy had paranodal remyelination in some teased nerve fibres, reflecting earlier paranodal demyelination. CONCLUSION: Experimental inoculation with C jejuni may provide a new model for understanding some forms of Guillain-Barré syndrome. Images PMID:8795599

  14. Effects of efflux-pump inducers and genetic variation of the multidrug transporter cmeB in biocide resistance of Campylobacter jejuni and Campylobacter coli.

    PubMed

    Mavri, Ana; Smole Možina, Sonja

    2013-03-01

    Multidrug efflux pumps, such as CmeABC and CmeDEF, are involved in the resistance of Campylobacter to a broad spectrum of antimicrobials. The aim of this study was to analyse the effects of two putative efflux-pump inducers, bile salts and sodium deoxycholate, on the resistance of Campylobacter to biocides (triclosan, benzalkonium chloride, chlorhexidine diacetate, cetylpyridinium chloride and trisodium phosphate), SDS and erythromycin. The involvement of the CmeABC and CmeDEF efflux pumps in this resistance was studied on the basis of the effects of bile salts and sodium deoxycholate in Campylobacter cmeB, cmeF and cmeR mutants. The genetic variation in the cmeB gene was also examined, to see whether this polymorphism is related to the function of the efflux pump. In 15 Campylobacter jejuni and 23 Campylobacter coli strains, bile salts and sodium deoxycholate increased the MICs of benzalkonium chloride, chlorhexidine diacetate, cetylpyridinium chloride and SDS, and decreased the MICs of triclosan, trisodium phosphate and erythromycin. Bile salts and sodium deoxycholate further decreased or increased the MICs of biocides and erythromycin in the cmeF and cmeR mutants. For cmeB polymorphisms, 17 different cmeB-specific PCR-RFLP patterns were identified: six within C. jejuni only, nine within C. coli only and two in both species. In conclusion, bile salts and sodium deoxycholate can increase or decrease bacterial resistance to structurally unrelated antimicrobials. The MIC increases in the cmeF and cmeR mutants indicated that at least one non-CmeABC efflux system is involved in resistance to biocides. These results indicate that the cmeB gene polymorphism identified is not associated with biocide and erythromycin resistance in Campylobacter.

  15. 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.

  16. Detection and initial characterization of novel capsular polysaccharide among diverse Campylobacter jejuni strains using alcian blue dye.

    PubMed

    Karlyshev, A V; Wren, B W

    2001-01-01

    We have recently demonstrated that most strains of Campylobacter jejuni produce capsular polysaccharide (CPS), which can be detected by immunoblotting with homologous Penner antisera on polyvinylidene difluoride membranes (A. V. Karlyshev, D. Linton, N. A. Gregson, A. J. Lastovica, and B. W. Wren, Mol. Microbiol. 35:529-541, 2000). In this report, we describe a universal and rapid staining procedure using Alcian blue for C. jejuni CPS, which does not rely on the availability of antisera and identifies CPS in untypeable strains. Furthermore, Alcian blue staining identified CPS in its lipid-free form directly on Tricine gels, and we demonstrate that CPS is thermostable and is accumulated in the culture supernatant in a lipid-free form. The identification of a newly described CPS and its lipid-free form in C. jejuni should prove invaluable in studying the pathogenesis and epidemiology of this important pathogen.

  17. Potential of coriander (Coriandrum sativum) oil as a natural antimicrobial compound in controlling Campylobacter jejuni in raw meat.

    PubMed

    Rattanachaikunsopon, Pongsak; Phumkhachorn, Parichat

    2010-01-01

    Twelve essential oils were tested in vitro for antimicrobial activities against several strains of Campylobacter jejuni, a pathogen causing food-borne diseases worldwide. Using disk diffusion and minimal inhibitory concentration determination assays, we noted that coriander oil exhibited the strongest antimicrobial activity against all tested strains. The oil had a bactericidal effect on the target bacteria. In evaluating the antimicrobial potency of coriander oil against C. jejuni on beef and chicken meat at 4 degrees C and 32 degrees C, it was found that the oil reduced the bacterial cell load in a dose-dependent manner. The type of meat and temperature did not influence the antimicrobial activity of the oil. This study indicates the potential of coriander oil to serve as a natural antimicrobial compound against C. jejuni in food.

  18. Updated Campylobacter jejuni Capsule PCR Multiplex Typing System and Its Application to Clinical Isolates from South and Southeast Asia

    PubMed Central

    Poly, Frédéric; Serichantalergs, Oralak; Kuroiwa, Janelle; Pootong, Piyarat; Mason, Carl; Guerry, Patricia; Parker, Craig T.

    2015-01-01

    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 the gold standard for C. jejuni typing. A preliminary multiplex PCR technique covering 17 serotypes was previously developed in order to replace this classic serotyping scheme. Here we report the completion of the multiplex PCR technology that is able to identify all the 47 Penner serotypes types known for C. jejuni. The number of capsule types represented within the 47 serotypes is 35. We have applied this method to a collection of 996 clinical isolates from Thailand, Cambodia and Nepal and were able to successfully determine capsule types of 98% of these. PMID:26630669

  19. Occupational Exposure to Swine, Poultry, and Cattle and Antibody Biomarkers of Campylobacter jejuni Exposure and Autoimmune Peripheral Neuropathy

    PubMed Central

    Vegosen, Leora; Breysse, Patrick N.; Agnew, Jacqueline; Gray, Gregory C.; Nachamkin, Irving; Sheikh, Kazim; Kamel, Freya; Silbergeld, Ellen

    2015-01-01

    Introduction Foodborne Campylobacter jejuni infection has been associated with an increased risk of autoimmune peripheral neuropathy, but risks of occupational exposure to C. jejuni have received less attention. This study compared anti-C. jejuni IgA, IgG, and IgM antibody levels, as well as the likelihood of testing positive for any of five anti-ganglioside autoantibodies, between animal farmers and non-farmers. Anti-C. jejuni antibody levels were also compared between farmers with different animal herd or flock sizes. The relationship between anti-C. jejuni antibody levels and detection of anti-ganglioside autoantibodies was also assessed. Methods Serum samples from 129 Agricultural Health Study swine farmers (some of whom also worked with other animals) and 46 non-farmers, all from Iowa, were analyzed for anti-C. jejuni antibodies and anti-ganglioside autoantibodies using ELISA. Information on animal exposures was assessed using questionnaire data. Anti-C. jejuni antibody levels were compared using Mann-Whitney tests and linear regression on log-transformed outcomes. Fisher’s Exact Tests and logistic regression were used to compare likelihood of positivity for anti-ganglioside autoantibodies. Results Farmers had significantly higher levels of anti-C. jejuni IgA (p < 0.0001) and IgG (p = 0.02) antibodies compared to non-farmers. There was no consistent pattern of anti-C. jejuni antibody levels based on animal herd or flock size. A higher percentage of farmers (21%) tested positive for anti-ganglioside autoantibodies compared to non-farmers (9%), but this difference was not statistically significant (p = 0.11). There was no significant association between anti-C. jejuni antibody levels and anti-ganglioside autoantibodies. Conclusions The findings provide evidence that farmers who work with animals may be at increased risk of exposure to C. jejuni. Future research should include longitudinal studies of exposures and outcomes, as well as studies of interventions

  20. Detection of heat-stable antigens of Campylobacter jejuni and C. coli by direct agglutination and passive hemagglutination.

    PubMed

    Oza, A N; Thwaites, R T; Wareing, D R A; Bolton, F J; Frost, J A

    2002-03-01

    The two serotyping schemes for the detection of heat-stable antigens of Campylobacter jejuni and Campylobacter coli use the same strains for antiserum production but differ in the detection systems used for identifying agglutination. The Penner method uses passive hemagglutination (PHA) while the Laboratory of Enteric Pathogens method uses the same antisera but in a whole-bacterial-cell direct agglutination (DA) protocol. C. jejuni produces a polysaccharide capsule, which is antigenic, and is the main component detected by the PHA method. The DA method will detect both capsule antigens and lipopolysaccharide (LPS) or lipooligosaccharide (LOS) surface antigens. Comparison of both methods by using a selection of isolates from human infection has shown a range of variation in agglutination specificity, reflecting the differences in antigens detected by the two methods. While 27.4% of the 416 C. jejuni isolates reacted with the antisera raised against the same type strains by either method, the majority showed a range of more complex relationships. None of the 37 C. coli isolates reacted with the same antiserum by both methods. Together the two schemes gave a total of 102 distinct combined serogroups for C. jejuni and 16 for C. coli. Thus, while some clonally related isolates share the same capsule and LOS or LPS antigens, other strains appear to have a common capsule antigen but differ in their LPS or LOS structures or vice versa.

  1. 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. PMID:25914692

  2. Important Role of a Putative Lytic Transglycosylase Cj0843c in β-Lactam Resistance in Campylobacter jejuni

    PubMed Central

    Zeng, Ximin; Gillespie, Barbara; Lin, Jun

    2015-01-01

    Beta-lactam antibiotics are an important class of antibiotics for treating bacterial infections. Despite prevalent β-lactam resistance in Campylobacter jejuni, the leading bacterial cause of human diarrhea in developed countries, molecular mechanism of β-lactam resistance in C. jejuni is still largely unknown. In this study, C. jejuni 81–176 was used for random transposon mutagenesis. Screening of a 2,800-mutant library identified 22 mutants with increased susceptibility to ampicillin. Of these mutants, two mutants contains mutations in Cj0843c (a putative lytic transglycosylase gene) and in its upstream gene Cj0844c, respectively. Complementation experiment demonstrated that the Cj0843 contributes to β-lactam resistance. The Cj0843c insertional mutation was subsequently introduced to diverse C. jejuni clinical strains for MIC test, showing that Cj0843c contributes to both intrinsic and acquired β-lactam resistance of C. jejuni. Consistent with this finding, inactivation of Cj0843c also dramatically reduced β-lactamase activity. Genomic examination and PCR analysis showed Cj0843c is widely distributed in C. jejuni. High purity recombinant Cj0843c was produced for generation of specific antiserum. The Cj0843 was localized in the periplasm, as demonstrated by immunoblotting using specific antibodies. Turbidimetric assay further demonstrated the capability of the purified Cj0843c to hydrolyze cell walls. Inactivation of Cj0843c also significantly reduced C. jejuni colonization in the intestine. Together, this study identifies a mechanism of β-lactam resistance in C. jejuni and provides insights into the role of cell wall metabolism in regulating β-lactamase activity. PMID:26635760

  3. Divergent distribution of the sensor kinase CosS in non-thermotolerant campylobacter species and its functional incompatibility with the response regulator CosR of Campylobacter jejuni.

    PubMed

    Hwang, Sunyoung; Miller, William G; Ryu, Sangryeol; Jeon, Byeonghwa

    2014-01-01

    Two-component signal transduction systems are commonly composed of a sensor histidine kinase and a cognate response regulator, modulating gene expression in response to environmental changes through a phosphorylation-dependent process. CosR is an OmpR-type response regulator essential for the viability of Campylobacter jejuni, a major foodborne pathogenic species causing human gastroenteritis. Although CosR is a response regulator, its cognate sensor kinase has not been identified in C. jejuni. In this study, DNA sequence analysis of the cosR flanking regions revealed that a gene encoding a putative sensor kinase, which we named cosS, is prevalent in non-thermotolerant Campylobacter spp., but not in thermotolerant campylobacters. Phosphorylation assays indicated that C. fetus CosS rapidly autophosphorylates and then phosphorylates C. fetus CosR, suggesting that the CosRS system constitutes a paired two-component signal transduction system in C. fetus. However, C. fetus CosS does not phosphorylate C. jejuni CosR, suggesting that CosR may have different regulatory cascades between thermotolerant and non-thermotolerant Campylobacter species. Comparison of CosR homolog amino acid sequences showed that the conserved phosphorylation residue (D51), which is present in all non-thermotolerant Campylobacter spp., is absent from the CosR homologs of thermotolerant Campylobacter species. However, C. jejuni CosR was not phosphorylated by C. fetus CosS even after site-directed mutagenesis of N51D, implying that C. jejuni CosR may possibly function phosphorylation-independently. In addition, the results of cosS mutational analysis indicated that CosS is not associated with the temperature dependence of the Campylobacter spp. despite its unique divergent distribution only in non-thermotolerant campylobacters. The findings in this study strongly suggest that thermotolerant and non-thermotolerant Campylobacter spp. have different signal sensing mechanisms associated with the Cos

  4. Serological analysis of the heat-stable antigens involved in serotyping Campylobacter jejuni and Campylobacter coli.

    PubMed

    Moran, A P; Kosunen, T U

    1989-03-01

    Analysis with serotyping antisera showed that carbohydrate determinants were the dominant heat-stable antigens of Campylobacter jenuni/coli involved, whereas proteins did not contribute to the serological reactions. Lipopolysaccharide (LPS) along with a polysaccharide extract from whole bacteria (PS(WB] conferred strain serospecificity. In general, analysis with monoclonal antibodies in passive haemagglutination and co-agglutination tests showed the existence of similar antigenic determinants in LPS and PS(WB) of the same strain. However, in some strains determinants were detectable in LPS but not in PS(WB) using monoclonal antibodies, in other strains the situation was reversed. All of these monoclonal antibodies reacted with LPS in the more sensitive immunoblotting technique. The presence of 2-keto-3-deoxyoctonic acid in PS(WB) preparations, in the absence of endotoxin, supported the conclusion that PS(WB) was derived from LPS during extraction. The lack of detection of a reaction by monoclonal antibodies with LPS in passive haemagglutination, in contrast to immunoblotting, was suggested due to the presence of low concentrations of the relevant epitopes because of the procedure used to prepare the LPS tested.

  5. Diversity and relatedness of Shiga toxin-producing Escherichia coli and Campylobacter jejuni between farms in a dairy catchment.

    PubMed

    Irshad, H; Cookson, A L; Ross, C M; Jaros, P; Prattley, D J; Donnison, A; McBRIDE, G; Marshall, J; French, N P

    2016-05-01

    The aim of this study was to examine the population structure, transmission and spatial relationship between genotypes of Shiga toxin-producing Escherichia coli (STEC) and Campylobacter jejuni, on 20 dairy farms in a defined catchment. Pooled faecal samples (n = 72) obtained from 288 calves were analysed by real-time polymerase chain reaction (rtPCR) for E. coli serotypes O26, O103, O111, O145 and O157. The number of samples positive for E. coli O26 (30/72) was high compared to E. coli O103 (7/72), O145 (3/72), O157 (2/72) and O111 (0/72). Eighteen E. coli O26 and 53 C. jejuni isolates were recovered from samples by bacterial culture. E. coli O26 and C. jejuni isolates were genotyped using pulsed-field gel electrophoresis and multilocus sequence typing, respectively. All E. coli O26 isolates could be divided into four clusters and the results indicated that E. coli O26 isolates recovered from calves on the same farm were more similar than isolates recovered from different farms in the catchment. There were 11 different sequence types of C. jejuni isolated from the cattle and 22 from water. An analysis of the population structure of C. jejuni isolated from cattle provided evidence of clustering of genotypes within farms, and among groups of farms separated by road boundaries.

  6. Double-locus sequence typing using porA and peb1A for epidemiological studies of Campylobacter jejuni.

    PubMed

    Ahmed, Monir U; Dunn, Louise; Valcanis, Mary; Hogg, Geoff; Ivanova, Elena P

    2014-03-01

    Campylobacter jejuni is the leading cause of foodborne bacterial gastroenteritis worldwide. Bacterial typing schemes play an important role in epidemiological investigations to trace the source and route of transmission of the infectious agent by identifying outbreak and differentiating among sporadic infections. In this study, a double-locus sequence typing (DLST) scheme for C. jejuni based on concatenated partial sequences of porA and peb1A genes is proposed. The DLST scheme was validated using 50 clinical and environmental C. jejuni strains isolated from human (C5, H, H15-H19), chicken (CH1-CH15), water (W2-W17), and ovine samples (OV1-OV6). The scheme was found to be highly discriminatory (discrimination index [DI]=0.964) and epidemiologically concordant based on C. jejuni strains studied. The DLST showed discriminatory power above 0.95 and excellent congruence to multilocus sequence typing and can be recommended as a rapid and low-cost typing scheme for epidemiological investigation of C. jejuni. It is suggested that the DLST scheme is suitable for identification of outbreak strains and differentiation of the sporadic infection strains.

  7. 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. PMID:26058384

  8. 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.

  9. Effect of high-temperature inside-outside spray on survival of campylobacter jejuni attached to prechill chicken carcasses.

    PubMed

    Li, Y; Yang, H; Swem, B L

    2002-09-01

    Prechill chicken carcasses, inoculated with Campylobacter jejuni, were sprayed in an inside-outside birdwasher at 20, 55, or 60 C, with or without 50 ppm chlorine, in a poultry processing pilot plant. Carcasses were sprayed for 12 s at 80 pounds per square inch (psi). Next, carcasses were placed in a chiller filled with 50 ppm chlorinated ice water at 4 C for 50 min. Most probable numbers of C. jejuni were determined based on chicken carcass wash water before and after the spray treatment. The skin color of chicken carcasses was measured. The results of this study showed that the 55 and 60 C water spray treatments significantly reduced C. jejuni by more than 0.78 log cfu/carcass compared with the 20 C water spray treatment. However, all of the 50 ppm chlorine spray treatments at three different temperatures were not significantly different. The skin color of chicken carcasses did not change significantly after the spray treatments at temperatures less than 60 C. The chilling process with 50 ppm chlorinated ice water at 4 C further reduced more C. jejuni (approximately 1 log cfu/carcass) among the water spray treatments but did not result in greater reduction of C. jejuni among the chlorine spray treatments.

  10. 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.

  11. Campylobacter infection

    MedlinePlus

    ... infection occurs in the small intestine from a bacteria called Campylobacter jejuni . It is a type of food poisoning. ... Campylobacter enteritis is a common cause of intestinal infection . ... of traveler's diarrhea or food poisoning . People most often ...

  12. Use of Direct LAMP Screening of Broiler Fecal Samples for Campylobacter jejuni and Campylobacter coli in the Positive Flock Identification Strategy

    PubMed Central

    Sabike, Islam I.; Uemura, Ryoko; Kirino, Yumi; Mekata, Hirohisa; Sekiguchi, Satoshi; Okabayashi, Tamaki; Goto, Yoshitaka; Yamazaki, Wataru

    2016-01-01

    Rapid identification of Campylobacter-positive flocks before slaughter, following freezing and heat treatment for the Campylobacter-positive carcasses at the slaughterhouses is an effective control strategy against foodborne campylobacteriosis. We evaluated a loop-mediated isothermal amplification (LAMP) assay for the direct screening of naturally contaminated chicken cloacal swabs for C. jejuni/C. coli to compare this assay with conventional quantitative culture methods. In a comparison study of 165 broilers, the LAMP assay showed 82.8% (48/58 by conventional culture) sensitivity, 100% (107/107) specificity, 100% (48/48) positive predictive value (PPV), and 91.5% (107/117) negative predictive value (NPV). In a comparison of 55 flocks, LAMP showed 90.5% (19/21) sensitivity, 100% (34/34) specificity, 100% (19/19) PPV, and 94.4% (34/36) NPV. In the cumulative total of 28 farm-level comparisons, LAMP showed 100% (12/12) sensitivity, 100% (16/16) specificity, 100% (12/12) PPV, and 100% (16/16) NPV. The LAMP assay required less than 90 min from the arrival of the fecal samples to final results in the laboratory. This suggests that the LAMP assay will facilitate the identification of C. jejuni/C. coli-positive broiler flocks at the farm level or in slaughterhouses before slaughtering, which would make it an effective tool in preventing the spread of Campylobacter contamination. PMID:27746777

  13. Escherichia coli O157:H7, Campylobacter jejuni, and Salmonella Prevalence in cull dairy cows marketed in northeastern Ohio.

    PubMed

    Dodson, Kathryn; LeJeune, Jeffrey

    2005-05-01

    Preharvest management factors are predicted to impact the prevalence of foodborne pathogens in cattle sent to slaughter. We simultaneously examined the prevalence and antibiotic resistance patterns of Campylobacter jejuni, Salmonella, and Escherichia coli O157:H7 isolated from cull dairy cattle at two livestock auctions in northeastern Ohio. Between April and September 2002, a total of 1,026 fecal samples were collected. C. jejuni was isolated from 48 of 686 (7%) fecal samples, Salmonella was isolated from 39 of 585 (6.7%) samples, and E. coli O157:H7 was isolated from 21 of 1,026 (2.1%) samples. Of the 585 samples tested for all three pathogens, at least one pathogen was identified in 86 of 585 (15%) samples. One sample was positive for both E. coli O157:H7 and C. jejuni, and five samples yielded both C. jejuni and Salmonella. Size of herd of origin could be traced for 75 to 85% of samples collected. Salmonella was isolated at higher frequencies from herds larger than 60 cattle than from smaller herds (9.0 versus 3.5%, P = 0.02). In contrast, size of herd of origin did not significantly affect the E. coli O157:H7 and C. jejuni prevalence. Approximately 90% of Salmonella and E. coli O157:H7 isolates were pansensitive to a panel of 16 antibiotics. Thirty-six percent of C. jejuni isolates were resistant to tetracycline. In this study, antibiotic resistance among the foodborne pathogens isolated from cull diary cattle was rare. Although size of dairy herd of origin was positively associated with Salmonella prevalence, herd size was not strongly associated with E. coli O157:H7 and C. jejuni prevalence in market dairy cattle. These results can be used to assess the food safety risks associated with the slaughter of cull dairy cattle.

  14. Structural characterization of Campylobacter jejuni lipooligosaccharide outer cores associated with Guillain-Barre and Miller Fisher syndromes.

    PubMed

    Godschalk, Peggy C R; Kuijf, Mark L; Li, Jianjun; St Michael, Frank; Ang, C Wim; Jacobs, Bart C; Karwaski, Marie-France; Brochu, Denis; Moterassed, Ali; Endtz, Hubert P; van Belkum, Alex; Gilbert, Michel

    2007-03-01

    Molecular mimicry between lipooligosaccharides (LOS) of Campylobacter jejuni and gangliosides in peripheral nerves plays a crucial role in the pathogenesis of C. jejuni-related Guillain-Barré syndrome (GBS). We have analyzed the LOS outer core structures of 26 C. jejuni strains associated with GBS and its variant, Miller Fisher syndrome (MFS), by capillary electrophoresis coupled with electrospray ionization mass spectrometry. Sixteen out of 22 (73%) GBS-associated and all 4 (100%) MFS-associated strains expressed LOS with ganglioside mimics. GM1a was the most prevalent ganglioside mimic in GBS-associated strains (10/22, 45%), and in eight of these strains, GM1a was found in combination with GD1a mimics. All seven strains isolated from patients with ophthalmoplegia (GBS or MFS) expressed disialylated (GD3 or GD1c) mimics. Three out of 22 GBS-associated strains (14%) did not express sialylated ganglioside mimics because their LOS locus lacked the genes necessary for sialylation. Three other strains (14%) did not express ganglioside mimics because of frameshift mutations in either the cstII sialyltransferase gene or the cgtB galactosyltransferase gene. It is not possible to determine if these mutations were already present during C. jejuni infection. This is the first report in which mass spectrometry combined with DNA sequence data were used to infer the LOS outer core structures of a large number of neuropathy-associated C. jejuni strains. We conclude that molecular mimicry between gangliosides and C. jejuni LOS is the presumable pathogenic mechanism in most cases of C. jejuni-related GBS. However, our findings suggest that in some cases, other mechanisms may play a role. Further examination of the disease etiology in these patients is mandatory.

  15. Effects of subtherapeutic administration of antimicrobial agents to beef cattle on the prevalence of antimicrobial resistance in Campylobacter jejuni and Campylobacter hyointestinalis.

    PubMed

    Inglis, G D; McAllister, T A; Busz, H W; Yanke, L J; Morck, D W; Olson, M E; Read, R R

    2005-07-01

    The influence of antimicrobial agents on the development of antimicrobial resistance (AMR) in Campylobacter isolates recovered from 300 beef cattle maintained in an experimental feedlot was monitored over a 315-day period (11 sample times). Groups of calves were assigned to one of the following antimicrobial treatments: chlortetracycline and sulfamethazine (CS), chlortetracycline alone (Ct), virginiamycin, monensin, tylosin phosphate, and no antimicrobial agent (i.e., control treatment). In total, 3,283 fecal samples were processed for campylobacters over the course of the experiment. Of the 2,052 bacterial isolates recovered, 92% were Campylobacter (1,518 were Campylobacter hyointestinalis and 380 were C. jejuni). None of the antimicrobial treatments decreased the isolation frequency of C. jejuni relative to the control treatment. In contrast, C. hyointestinalis was isolated less frequently from animals treated with CS and to a lesser extent from animals treated with Ct. The majority (> or =94%) of C. jejuni isolates were sensitive to ampicillin, erythromycin, and ciprofloxacin, but more isolates with resistance to tetracycline were recovered from animals fed Ct. All of the 1,500 isolates of C. hyointestinalis examined were sensitive to ciprofloxacin. In contrast, 11%, 10%, and 1% of these isolates were resistant to tetracycline, erythromycin, and ampicillin, respectively. The number of animals from which C. hyointestinalis isolates with resistance to erythromycin and tetracycline were recovered differed among the antimicrobial treatments. Only Ct administration increased the carriage rates of erythromycin-resistant isolates of C. hyointestinalis, and the inclusion of CS in the diet increased the number of animals from which tetracycline-resistant isolates were recovered. The majority of C. hyointestinalis isolates with resistance to tetracycline were obtained from cohorts within a single pen, and most of these isolates were recovered from cattle during feeding of

  16. Rapid Identification of Novel Immunodominant Proteins and Characterization of a Specific Linear Epitope of Campylobacter jejuni

    PubMed Central

    Hoppe, Sebastian; Bier, Frank F.; Nickisch-Rosenegk, Markus v.

    2013-01-01

    Campylobacter jejuni remains one of the major gut pathogens of our time. Its zoonotic nature and wide-spread distribution in industrialized countries calls for a quick and reliable diagnostic tool. Antibody-based detection presents a suitable means to identify pathogenic bacteria. However, the knowledge about immunodominant targets is limited. Thus, an approach is presented, which allows for the rapid screening of numerous cDNA derived expression clones to identify novel antigens. The deeper understanding of immunodominant proteins assists in the design of diagnostic tools and furthers the insight into the bacterium’s pathogenicity as well as revealing potential candidates for vaccination. We have successfully screened 1536 clones of an expression library to identify 22 proteins that have not been described as immunodominant before. After subcloning the corresponding 22 genes and expression of full-length proteins, we investigated the immunodominant character by microarrays and ELISA. Subsequently, seven proteins were selected for epitope mapping. For cj0669 and cj0920c linear epitopes were identified. For cj0669, specificity assays revealed a specific linear epitope site. Consequently, an eleven amino acid residue sequence TLIKELKRLGI was analyzed via alanine scan, which revealed the glycine residue to be significant for binding of the antibody. The innovative approach presented herein of generating cDNAs of prokaryotes in combination with a microarray platform rendering time-consuming purification steps obsolete has helped to illuminate novel immunodominant proteins of C.jejuni. The findings of a specific linear epitope pave the way for a plethora of future research and the potential use in diagnostic applications such as serological screenings. Moreover, the current approach is easily adaptable to other highly relevant bacteria making it a formidable tool for the future discovery of antigens and potential biomarkers. Consequently, it is desirable to simplify

  17. Rapid identification of novel immunodominant proteins and characterization of a specific linear epitope of Campylobacter jejuni.

    PubMed

    Hoppe, Sebastian; Bier, Frank F; von Nickisch-Rosenegk, Markus; Nickisch-Rosenegk, Markus V

    2013-01-01

    Campylobacter jejuni remains one of the major gut pathogens of our time. Its zoonotic nature and wide-spread distribution in industrialized countries calls for a quick and reliable diagnostic tool. Antibody-based detection presents a suitable means to identify pathogenic bacteria. However, the knowledge about immunodominant targets is limited. Thus, an approach is presented, which allows for the rapid screening of numerous cDNA derived expression clones to identify novel antigens. The deeper understanding of immunodominant proteins assists in the design of diagnostic tools and furthers the insight into the bacterium's pathogenicity as well as revealing potential candidates for vaccination. We have successfully screened 1536 clones of an expression library to identify 22 proteins that have not been described as immunodominant before. After subcloning the corresponding 22 genes and expression of full-length proteins, we investigated the immunodominant character by microarrays and ELISA. Subsequently, seven proteins were selected for epitope mapping. For cj0669 and cj0920c linear epitopes were identified. For cj0669, specificity assays revealed a specific linear epitope site. Consequently, an eleven amino acid residue sequence TLIKELKRLGI was analyzed via alanine scan, which revealed the glycine residue to be significant for binding of the antibody. The innovative approach presented herein of generating cDNAs of prokaryotes in combination with a microarray platform rendering time-consuming purification steps obsolete has helped to illuminate novel immunodominant proteins of C.jejuni. The findings of a specific linear epitope pave the way for a plethora of future research and the potential use in diagnostic applications such as serological screenings. Moreover, the current approach is easily adaptable to other highly relevant bacteria making it a formidable tool for the future discovery of antigens and potential biomarkers. Consequently, it is desirable to simplify the

  18. Complete 6-Deoxy-d-altro-heptose Biosynthesis Pathway from Campylobacter jejuni

    PubMed Central

    McCallum, Matthew; Shaw, Steven D.; Shaw, Gary S.; Creuzenet, Carole

    2012-01-01

    The Campylobacter jejuni capsule is important for colonization and virulence in various infection models. In most strains, the capsule includes a modified heptose whose biological role and biosynthetic pathway are unknown. To decipher the biosynthesis pathway for the 6-deoxy-d-altro-heptose of strain 81-176, we previously showed that the 4,6-dehydratase WcbK and the reductase WcaG generated GDP-6-deoxy-d-manno-heptose, but the C3 epimerase necessary to form GDP-6-deoxy-d-altro-heptose was not identified. Herein, we characterized the putative C3/C5 epimerase Cjj1430 and C3/C5 epimerase/C4 reductase Cjj1427 from the capsular cluster. We demonstrate that GDP-6-deoxy-d-altro-heptose biosynthesis is more complex than anticipated and requires the sequential action of WcbK, Cjj1430, and Cjj1427. We show that Cjj1430 serves as C3 epimerase devoid of C5 epimerization activity and that Cjj1427 has no epimerization activity and only serves as a reductase to produce GDP-6-deoxy-d-altro-heptose. Cjj1430 and Cjj1427 are the only members of the C3/C5 epimerases and C3/C5 epimerase/C4 reductase families shown to have activity on a heptose substrate and to exhibit only one of their two to three potential activities, respectively. Furthermore, we show that although the reductase WcaG is not part of the main pathway, its presence and its product affect the outcome of the pathway in a complex regulatory loop involving Cjj1427. This work provides the grounds for the elucidation of similar pathways found in other C. jejuni strains and other pathogens. It provides new molecular tools for the synthesis of carbohydrate antigens useful for vaccination and for the screening of enzymatic inhibitors that may have antibacterial effects. PMID:22787156

  19. Serological diversity and chemical structures of Campylobacter jejuni low-molecular-weight lipopolysaccharides.

    PubMed Central

    Aspinall, G O; McDonald, A G; Raju, T S; Pang, H; Mills, S D; Kurjanczyk, L A; Penner, J L

    1992-01-01

    Low-Mr lipopolysaccharides (LPS) of Campylobacter jejuni reference strains for serotypes O:1, O:4, O:23, and O:36 were examined through the liberation of core oligosaccharides by mild acid cleavage of the ketosidic linkage of 3-deoxy-D-manno-2-octulosonic acid residues to the lipid A moiety. The liberated oligosaccharides were examined for chemical structure by compositional analysis and methylated linkage analysis in conjunction with fast atom bombardment-mass spectrometry of permethylated oligosaccharide derivatives. The results showed (i) that the LPS contained short oligosaccharide chains of branched nonrepetitive structure, to many of which N-acetylneuraminic acid residues remained attached by 2----3 linkages to 4-linked D-galactose residues in the core structure; (ii) that serotypical differences, which are not readily defined through qualitatively similar compositions, are clearly reflected in variations in linkage types and sequences of sugar residues in the outer core attached to an inner region of invariable structure; but (iii) that the presence or absence of NeuAc residues does not appear to be a basis for serotypical differences. The results also showed that oligosaccharide chains from LPS of serotypes O:1 and O:4 are distinctly different and are distinct again from those of the cross-reacting serotypes O:23 and O:36, between whose core oligosaccharide chains no differences were found. It is concluded that the structurally variable low-Mr LPS from C. jejuni show greater similarities to the lipooligosaccharides from Neisseria spp. than to the highly conserved core regions of Salmonella species. Those strains (serotypes O:23 and O:36) which also furnish high-Mr LPS are unique among gram-negative bacteria in possessing both low-Mr molecules of the Neisseria lipooligosaccharide type and high-Mr LPS of the Salmonella smooth type. Images PMID:1370951

  20. Tracing Back Clinical Campylobacter jejuni in the Northwest of Italy and Assessing Their Potential Source.

    PubMed

    Di Giannatale, Elisabetta; Garofolo, Giuliano; Alessiani, Alessandra; Di Donato, Guido; Candeloro, Luca; Vencia, Walter; Decastelli, Lucia; Marotta, Francesca

    2016-01-01

    Food-borne campylobacteriosis is caused mainly by the handling or consumption of undercooked chicken meat or by the ingestion of contaminated raw milk. Knowledge about the contributions of different food sources to gastrointestinal disease is fundamental to prioritize food safety interventions and to establish proper control strategies. Assessing the genetic diversity among Campylobacter species is essential to our understanding of their epidemiology and population structure. We molecularly characterized 56 Campylobacter jejuni isolates (31 from patients hospitalized with gastroenteritis, 17 from raw milk samples, and 8 from chicken samples) using multilocus sequence typing (MLST) and pulsed-field gel electrophoresis (PFGE) in order to trace the source of the disease. We also used a population genetic approach to investigate the source of the human cases from six different reservoirs of infection. MLST identified 25 different sequence types and 11 clonal complexes (CCs) (21, 658, 206, 353, 443, 48, 61, 257, 1332, 354, 574) and these included several alleles not cited previously in the PubMLST international database. The most prevalent CCs were 21, 206, and 354. PFGE showed 34 pulsotypes divided between 28 different clusters. At the fine scale, by means of PFGE and MLST, only two human cases were linked to raw milk, while one case was linked to chicken meat. The investigation revealed the presence of several genotypes among the human isolates, which probably suggests multiple foci for the infections. Finally, the source attribution model we used revealed that most cases were attributed to chicken (69.75%) as the main reservoir in Italy, followed to a lesser extent by the following sources: cattle (8.25%); environment (6.28%); wild bird (7.37%); small ruminant (5.35%), and pork (2.98%). This study confirms the importance of correlating epidemiological investigations with molecular epidemiological data to better understand the dynamics of infection. PMID:27379033

  1. Tracing Back Clinical Campylobacter jejuni in the Northwest of Italy and Assessing Their Potential Source

    PubMed Central

    Di Giannatale, Elisabetta; Garofolo, Giuliano; Alessiani, Alessandra; Di Donato, Guido; Candeloro, Luca; Vencia, Walter; Decastelli, Lucia; Marotta, Francesca

    2016-01-01

    Food-borne campylobacteriosis is caused mainly by the handling or consumption of undercooked chicken meat or by the ingestion of contaminated raw milk. Knowledge about the contributions of different food sources to gastrointestinal disease is fundamental to prioritize food safety interventions and to establish proper control strategies. Assessing the genetic diversity among Campylobacter species is essential to our understanding of their epidemiology and population structure. We molecularly characterized 56 Campylobacter jejuni isolates (31 from patients hospitalized with gastroenteritis, 17 from raw milk samples, and 8 from chicken samples) using multilocus sequence typing (MLST) and pulsed-field gel electrophoresis (PFGE) in order to trace the source of the disease. We also used a population genetic approach to investigate the source of the human cases from six different reservoirs of infection. MLST identified 25 different sequence types and 11 clonal complexes (CCs) (21, 658, 206, 353, 443, 48, 61, 257, 1332, 354, 574) and these included several alleles not cited previously in the PubMLST international database. The most prevalent CCs were 21, 206, and 354. PFGE showed 34 pulsotypes divided between 28 different clusters. At the fine scale, by means of PFGE and MLST, only two human cases were linked to raw milk, while one case was linked to chicken meat. The investigation revealed the presence of several genotypes among the human isolates, which probably suggests multiple foci for the infections. Finally, the source attribution model we used revealed that most cases were attributed to chicken (69.75%) as the main reservoir in Italy, followed to a lesser extent by the following sources: cattle (8.25%); environment (6.28%); wild bird (7.37%); small ruminant (5.35%), and pork (2.98%). This study confirms the importance of correlating epidemiological investigations with molecular epidemiological data to better understand the dynamics of infection. PMID:27379033

  2. 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.

  3. Point mutations in the major outer membrane protein drive hypervirulence of a rapidly expanding clone of Campylobacter jejuni.

    PubMed

    Wu, Zuowei; Periaswamy, Balamurugan; Sahin, Orhan; Yaeger, Michael; Plummer, Paul; Zhai, Weiwei; Shen, Zhangqi; Dai, Lei; Chen, Swaine L; Zhang, Qijing

    2016-09-20

    Infections due to clonal expansion of highly virulent bacterial strains are clear and present threats to human and animal health. Association of genetic changes with disease is now a routine, but identification of causative mutations that enable disease remains difficult. Campylobacter jejuni is an important zoonotic pathogen transmitted to humans mainly via the foodborne route. C. jejuni typically colonizes the gut, but a hypervirulent and rapidly expanding clone of C. jejuni recently emerged, which is able to translocate across the intestinal tract, causing systemic infection and abortion in pregnant animals. The genetic basis responsible for this hypervirulence is unknown. Here, we developed a strategy, termed "directed genome evolution," by using hybridization between abortifacient and nonabortifacient strains followed by selection in an animal disease model and whole-genome sequence analysis. This strategy successfully identified SNPs in porA, encoding the major outer membrane protein, are responsible for the hypervirulence. Defined mutagenesis verified that these mutations were both necessary and sufficient for causing abortion. Furthermore, sequence analysis identified porA as the gene with the top genome-wide signal of adaptive evolution using Fu's Fs, a population genetic metric for recent population size changes, which is consistent with the recent expansion of clone "sheep abortion." These results identify a key virulence factor in Campylobacter and a potential target for the control of this zoonotic pathogen. Furthermore, this study provides general, unbiased experimental and computational approaches that are broadly applicable for efficient elucidation of disease-causing mutations in bacterial pathogens. PMID:27601641

  4. PCR-Restriction Fragment Length Polymorphism Analysis of Campylobacter jejuni Genes Involved in Lipooligosaccharide Biosynthesis Identifies Putative Molecular Markers for Guillain-Barré Syndrome▿

    PubMed Central

    Godschalk, Peggy C. R.; van Belkum, Alex; van den Braak, Nicole; van Netten, Diana; Ang, C. Wim; Jacobs, Bart C.; Gilbert, Michel; Endtz, Hubert P.

    2007-01-01

    Molecular mimicry of Campylobacter jejuni lipooligosaccharides (LOS) by gangliosides in peripheral nerve tissue probably triggers the Guillain-Barré syndrome due to the induction of cross-reactive antibodies. PCR-restriction fragment length polymorphism analysis of C. jejuni genes involved in the biosynthesis of LOS demonstrated that specific genes were associated with the expression of ganglioside mimics and the development of neuropathy. PMID:17507514

  5. In vitro susceptibility of Campylobacter jejuni to 27 antimicrobial agents and various combinations of beta-lactams with clavulanic acid or sulbactam.

    PubMed Central

    Van der Auwera, P; Scorneaux, B

    1985-01-01

    The in vitro susceptibility of human isolates of Campylobacter jejuni was investigated with 27 antibiotics and 8 combinations of beta-lactams with clavulanic acid or sulbactam. Ansamycin, the new quinolines, erythromycin, and cefpirome were the most active drugs against C. jejuni; amoxicillin, ampicillin, cefotaxime, and ceftazidime 90% of the isolates, greater than or equal to 50 mg/liter). The activity of various beta-lactams was unchanged by the addition of clavulanic acid or sulbactam. PMID:2994557

  6. Genetic Characteristics and Multiple-PCR Development for Capsular Identification of Specific Serotypes of Campylobacter jejuni

    PubMed Central

    Liang, Hao; Zhang, Aiyu; Gu, Yixin; You, Yuanhai; Zhang, Jianzhong; Zhang, Maojun

    2016-01-01

    The polysaccharide capsule (CPS) of Campylobacter jejuni is a virulence factor linked to cell surface carbohydrate diversity which mainly determines the serotypes. Thirty-four CPS gene cluster structures have been published and some of them can be distinguished by multiple-PCR. Penner serotypes HS1/44c, HS2, HS4c, HS19, HS23/36c and HS41 are markers for Guillain—Barré syndrome (GBS). The capsules may contribute to GBS susceptibility. Analysis of 18 CPS loci revealed high gene content diversity and a mosaic nature of the capsule loci, which are possibly due to gene gain/loss events, and demonstrated a high degree of conservation of genes within serotypes/serotype complexes. A method of multiple-PCR was developed to distinguish five specific serotypes and three GBS-related serotypes. Primers specific for each capsule type were designed on the basis of paralogs or a unique DNA region of the CPS locus. The multiple-PCR can distinguish the eight serotypes in two PCRs with sensitivity and specificity of 100% using 227 strains of known Penner type. The multiple-PCR method will help to distinguish serotypes simply and rapidly. PMID:27788180

  7. Structure of anabolic ornithine carbamoyltransferase from Campylobacter jejuni at 2.7 Å resolution

    PubMed Central

    Shabalin, I. G.; Porebski, P. J.; Cooper, D. R.; Grabowski, M.; Onopriyenko, O.; Grimshaw, S.; Savchenko, A.; Chruszcz, M.; Minor, W.

    2012-01-01

    Anabolic ornithine transcarbamoylase (aOTC) catalyzes the reaction between carbamoyl phosphate (CP) and l-ornithine (ORN) to form l-citrulline and phosphate in the urea cycle and l-arginine biosynthesis. The crystal structure of unliganded aOTC from Campylobacter jejuni (Cje aOTC) was determined at 2.7 Å resolution and refined to an R work of 20.3% and an R free of 24.0%. Cje aOTC is a trimer that forms a head-to-head pseudohexamer in the asymmetric unit. Each monomer is composed of an N-terminal CP-binding domain and a C-­terminal ORN-binding domain joined by two interdomain helices. The Cje aOTC structure presents an open conformation of the enzyme with a relatively flexible orientation of the ORN-binding domain respective to the CP-binding domain. The conformation of the B2–H3 loop (residues 68–78), which is involved in binding CP in an adjacent subunit of the trimer, differs from that seen in homologous proteins with CP bound. The loop containing the ORN-binding motif (DxxxSMG, residues 223–230) has a conformation that is different from those observed in unliganded OTC structures from other species, but is similar to those in structures with bound ORN analogs. The major differences in tertiary structure between Cje aOTC and human aOTC are described. PMID:22949186

  8. Crystal Structure and Catalytic Mechanism of PglD from Campylobacter jejuni

    SciTech Connect

    Olivier, N.; Imperiali, B

    2008-01-01

    The carbohydrate 2, 4-diacetamido-2, 4, 6-trideoxy-{alpha}-d-glucopyranose (BacAc2) is found in a variety of eubacterial pathogens. In Campylobacter jejuni, PglD acetylates the C4 amino group on UDP-2-acetamido-4-amino-2, 4, 6-trideoxy-a-d-glucopyranose (UDP-4-amino-sugar) to form UDP-BacAc2. Sequence analysis predicts PglD to be a member of the left-handed {Beta} helix family of enzymes. However, poor sequence homology between PglD and left-handed {Beta} helix enzymes with existing structural data precludes unambiguous identification of the active site. The co-crystal structures of PglD in the presence of citrate, acetyl coenzyme A, or the UDP-4-amino-sugar were solved. The biological assembly is a trimer with one active site formed between two protomers. Residues lining the active site were identified, and results from functional assays on alanine mutants suggest His-125 is critical for catalysis, whereas His-15 and His-134 are involved in substrate binding. These results are discussed in the context of implications for proteins homologous to PglD in other pathogens.

  9. A novel DNA-binding protein from Campylobacter jejuni bacteriophage NCTC12673.

    PubMed

    Arutyunov, Denis; Szymanski, Christine M

    2015-11-01

    We previously suggested that the double-stranded genomic DNA of Campylobacter jejuni bacteriophage NCTC12673 was complexed with proteins. Mass spectrometry of peptides obtained from tryptic digests of purified phage DNA indicated that phage protein Gp001 co-purified with the DNA. Gp001 is an acidic protein that lacks any obvious homology or conserved domains found in known DNA-binding proteins. The DNA-binding ability of recombinant Gp001 was examined using an electrophoretic mobility shift assay. Slow DNA-Gp001 complex formation was observed at pH 5.5, but not at neutral or basic pH. This nucleoprotein complex had difficulty entering agarose gels used in the assay while proteinase K pretreatment released the DNA from the complex. No mobility shift was observed when the DNA was immediately subjected to electrophoresis after mixing with Gp001, even if both components were separately pre-incubated at pH 5.5. The complexed DNA was unable to transform chemically competent Escherichia coli cells and was less susceptible to degradation by nucleases. The formation of Gp001-DNA complexes at low pH may provide a mechanism for maintaining DNA integrity while the phage pursues its host through the gastrointestinal tract. Also, this feature can potentially be used to improve DNA delivery protocols applied in gene therapy.

  10. Gene function hypotheses for the Campylobacter jejuni glycome generated by a logic-based approach.

    PubMed

    Sternberg, Michael J E; Tamaddoni-Nezhad, Alireza; Lesk, Victor I; Kay, Emily; Hitchen, Paul G; Cootes, Adrian; van Alphen, Lieke B; Lamoureux, Marc P; Jarrell, Harold C; Rawlings, Christopher J; Soo, Evelyn C; Szymanski, Christine M; Dell, Anne; Wren, Brendan W; Muggleton, Stephen H

    2013-01-01

    Increasingly, experimental data on biological systems are obtained from several sources and computational approaches are required to integrate this information and derive models for the function of the system. Here, we demonstrate the power of a logic-based machine learning approach to propose hypotheses for gene function integrating information from two diverse experimental approaches. Specifically, we use inductive logic programming that automatically proposes hypotheses explaining the empirical data with respect to logically encoded background knowledge. We study the capsular polysaccharide biosynthetic pathway of the major human gastrointestinal pathogen Campylobacter jejuni. We consider several key steps in the formation of capsular polysaccharide consisting of 15 genes of which 8 have assigned function, and we explore the extent to which functions can be hypothesised for the remaining 7. Two sources of experimental data provide the information for learning-the results of knockout experiments on the genes involved in capsule formation and the absence/presence of capsule genes in a multitude of strains of different serotypes. The machine learning uses the pathway structure as background knowledge. We propose assignments of specific genes to five previously unassigned reaction steps. For four of these steps, there was an unambiguous optimal assignment of gene to reaction, and to the fifth, there were three candidate genes. Several of these assignments were consistent with additional experimental results. We therefore show that the logic-based methodology provides a robust strategy to integrate results from different experimental approaches and propose hypotheses for the behaviour of a biological system.

  11. Identification of Campylobacter jejuni Genes Involved in the Response to Acidic pH and Stomach Transit▿

    PubMed Central

    Reid, Anne N.; Pandey, Reenu; Palyada, Kiran; Naikare, Hemant; Stintzi, Alain

    2008-01-01

    Campylobacter jejuni causes food- and waterborne gastroenteritis, and as such it must survive passage through the stomach in order to reach the gastrointestinal tract. While little is known about how C. jejuni survives transit through the stomach, its low infectious dose suggests it is well equipped to sense and respond to acid shock. In this study, the transcriptional profile of C. jejuni NCTC 11168 was obtained after the organism was exposed to in vitro and in vivo (piglet stomach) acid shock. The observed down-regulation of genes encoding ribosomal proteins likely reflects the need to reshuffle energy toward the expression of components required for survival. Acid shock also caused C. jejuni to up-regulate genes involved in stress responses. These included heat shock genes as well as genes involved in the response to oxidative and nitrosative stress. A role for the chaperone clpB in acid resistance was confirmed in vitro. Some genes showed expression patterns that were markedly different in vivo and in vitro, which likely reflects the complexity of the in vivo environment. For instance, transit through the stomach was characterized by up-regulation of genes that encode products that are involved in the use of nitrite as a terminal electron acceptor and down-regulation of genes that are involved in capsular polysaccharide expression. In conclusion, this study has enabled us to understand how C. jejuni modulates gene expression in response to acid shock in vitro and to correlate this with gene expression profiles of C. jejuni as it transits through the host stomach. PMID:18192414

  12. High Throughput Method for Analysis of Repeat Number for 28 Phase Variable Loci of Campylobacter jejuni Strain NCTC11168

    PubMed Central

    Lango-Scholey, Lea; Aidley, Jack; Woodacre, Alexandra; Jones, Michael A.

    2016-01-01

    Mutations in simple sequence repeat tracts are a major mechanism of phase variation in several bacterial species including Campylobacter jejuni. Changes in repeat number of tracts located within the reading frame can produce a high frequency of reversible switches in gene expression between ON and OFF states. The genome of C. jejuni strain NCTC11168 contains 29 loci with polyG/polyC tracts of seven or more repeats. This protocol outlines a method—the 28-locus-CJ11168 PV-analysis assay—for rapidly determining ON/OFF states of 28 of these phase-variable loci in a large number of individual colonies from C. jejuni strain NCTC11168. The method combines a series of multiplex PCR assays with a fragment analysis assay and automated extraction of fragment length, repeat number and expression state. This high throughput, multiplex assay has utility for detecting shifts in phase variation states within and between populations over time and for exploring the effects of phase variation on adaptation to differing selective pressures. Application of this method to analysis of the 28 polyG/polyC tracts in 90 C. jejuni colonies detected a 2.5-fold increase in slippage products as tracts lengthened from G8 to G11 but no difference between tracts of similar length indicating that flanking sequence does not influence slippage rates. Comparison of this observed slippage to previously measured mutation rates for G8 and G11 tracts in C. jejuni indicates that PCR amplification of a DNA sample will over-estimate phase variation frequencies by 20-35-fold. An important output of the 28-locus-CJ11168 PV-analysis assay is combinatorial expression states that cannot be determined by other methods. This method can be adapted to analysis of phase variation in other C. jejuni strains and in a diverse range of bacterial species. PMID:27466808

  13. High Throughput Method for Analysis of Repeat Number for 28 Phase Variable Loci of Campylobacter jejuni Strain NCTC11168.

    PubMed

    Lango-Scholey, Lea; Aidley, Jack; Woodacre, Alexandra; Jones, Michael A; Bayliss, Christopher D

    2016-01-01

    Mutations in simple sequence repeat tracts are a major mechanism of phase variation in several bacterial species including Campylobacter jejuni. Changes in repeat number of tracts located within the reading frame can produce a high frequency of reversible switches in gene expression between ON and OFF states. The genome of C. jejuni strain NCTC11168 contains 29 loci with polyG/polyC tracts of seven or more repeats. This protocol outlines a method-the 28-locus-CJ11168 PV-analysis assay-for rapidly determining ON/OFF states of 28 of these phase-variable loci in a large number of individual colonies from C. jejuni strain NCTC11168. The method combines a series of multiplex PCR assays with a fragment analysis assay and automated extraction of fragment length, repeat number and expression state. This high throughput, multiplex assay has utility for detecting shifts in phase variation states within and between populations over time and for exploring the effects of phase variation on adaptation to differing selective pressures. Application of this method to analysis of the 28 polyG/polyC tracts in 90 C. jejuni colonies detected a 2.5-fold increase in slippage products as tracts lengthened from G8 to G11 but no difference between tracts of similar length indicating that flanking sequence does not influence slippage rates. Comparison of this observed slippage to previously measured mutation rates for G8 and G11 tracts in C. jejuni indicates that PCR amplification of a DNA sample will over-estimate phase variation frequencies by 20-35-fold. An important output of the 28-locus-CJ11168 PV-analysis assay is combinatorial expression states that cannot be determined by other methods. This method can be adapted to analysis of phase variation in other C. jejuni strains and in a diverse range of bacterial species. PMID:27466808

  14. Whole Genome Comparison of Campylobacter jejuni Human Isolates Using a Low-Cost Microarray Reveals Extensive Genetic Diversity

    PubMed Central

    Dorrell, Nick; Mangan, Joseph A.; Laing, Kenneth G.; Hinds, Jason; Linton, Dennis; Al-Ghusein, Hasan; Barrell, Bart G.; Parkhill, Julian; Stoker, Neil G.; Karlyshev, Andrey V.; Butcher, Philip D.; Wren, Brendan W.

    2001-01-01

    Campylobacter jejuni is the leading cause of bacterial food-borne diarrhoeal disease throughout the world, and yet is still a poorly understood pathogen. Whole genome microarray comparisons of 11 C. jejuni strains of diverse origin identified genes in up to 30 NCTC 11168 loci ranging from 0.7 to 18.7 kb that are either absent or highly divergent in these isolates. Many of these regions are associated with the biosynthesis of surface structures including flagella, lipo-oligosaccharide, and the newly identified capsule. Other strain-variable genes of known function include those responsible for iron acquisition, DNA restriction/modification, and sialylation. In fact, at least 21% of genes in the sequenced strain appear dispensable as they are absent or highly divergent in one or more of the isolates tested, thus defining 1300 C. jejuni core genes. Such core genes contribute mainly to metabolic, biosynthetic, cellular, and regulatory processes, but many virulence determinants are also conserved. Comparison of the capsule biosynthesis locus revealed conservation of all the genes in this region in strains with the same Penner serotype as strain NCTC 11168. By contrast, between 5 and 17 NCTC 11168 genes in this region are either absent or highly divergent in strains of a different serotype from the sequenced strain, providing further evidence that the capsule accounts for Penner serotype specificity. These studies reveal extensive genetic diversity among C. jejuni strains and pave the way toward identifying correlates of pathogenicity and developing improved epidemiological tools for this problematic pathogen. PMID:11591647

  15. Campylobacter jejuni fatty acid synthase II: Structural and functional analysis of [beta]-hydroxyacyl-ACP dehydratase (FabZ)

    SciTech Connect

    Kirkpatrick, Andrew S.; Yokoyama, Takeshi; Choi, Kyoung-Jae; Yeo, Hye-Jeong

    2009-08-14

    Fatty acid biosynthesis is crucial for all living cells. In contrast to higher organisms, bacteria use a type II fatty acid synthase (FAS II) composed of a series of individual proteins, making FAS II enzymes excellent targets for antibiotics discovery. The {beta}-hydroxyacyl-ACP dehydratase (FabZ) catalyzes an essential step in the FAS II pathway. Here, we report the structure of Campylobacter jejuni FabZ (CjFabZ), showing a hexamer both in crystals and solution, with each protomer adopting the characteristic hot dog fold. Together with biochemical analysis of CjFabZ, we define the first functional FAS II enzyme from this pathogen, and provide a framework for investigation on roles of FAS II in C. jejuni virulence

  16. 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

  17. Campylobacter jejuni Increases Flagellar Expression and Adhesion of Noninvasive Escherichia coli: Effects on Enterocytic Toll-Like Receptor 4 and CXCL-8 Expression

    PubMed Central

    Reti, Kristen L.; Tymensen, Lisa D.; Davis, Shevaun P.; Amrein, Matthias W.

    2015-01-01

    Campylobacter jejuni is the most common cause of bacterium-induced gastroenteritis, and while typically self-limiting, C. jejuni infections are associated with postinfectious intestinal disorders, including flares in patients with inflammatory bowel disease and postinfectious irritable bowel syndrome (PI-IBS), via mechanisms that remain obscure. Based on the hypothesis that acute campylobacteriosis may cause pathogenic microbiota dysbiosis, we investigated whether C. jejuni may activate dormant virulence genes in noninvasive Escherichia coli and examined the epithelial pathophysiological consequences of these alterations. Microarray and quantitative real-time PCR analyses revealed that E. coli adhesin, flagellum, and hemolysin gene expression were increased when E. coli was exposed to C. jejuni-conditioned medium. Increased development of bacterial flagella upon exposure to live C. jejuni or C. jejuni-conditioned medium was observed under transmission electron microscopy. Atomic force microscopy demonstrated that the forces of bacterial adhesion to colonic T84 enterocytes, and the work required to rupture this adhesion, were significantly increased in E. coli exposed to C. jejuni-conditioned media. Finally, C. jejuni-modified E. coli disrupted TLR4 gene expression and induced proinflammatory CXCL-8 gene expression in colonic enterocytes. Together, these data suggest that exposure to live C. jejuni, and/or to its secretory-excretory products, may activate latent virulence genes in noninvasive E. coli and that these alterations may directly trigger proinflammatory signaling in intestinal epithelia. These observations shed new light on mechanisms that may contribute, at least in part, to postcampylobacteriosis inflammatory disorders. PMID:26371123

  18. Outbreak of Campylobacter jejuni infections associated with drinking unpasteurized milk procured through a cow-leasing program--Wisconsin, 2001.

    PubMed

    2002-06-28

    On December 7, 2001, the Sawyer County Department of Health and Human Services in northwestern Wisconsin notified the Wisconsin Division of Public Health about five cases of Campylobacter jejuni enteritis. All of the ill persons drank unpasteurized milk obtained at a local dairy farm. This report summarizes the investigation of these and other cases and of a cow-leasing program used to circumvent regulations prohibiting the sale of unpasteurized milk in Wisconsin. The outbreak highlights the hazards of consuming unpasteurized milk and milk products.

  19. [Antimicrobial Susceptibility and Resistance Mutations in Campylobacter jejuni and C. coli Isolates from Human and Meat Sources].

    PubMed

    Oishi, Akira; Murakami, Koichi; Etoh, Yoshiki; Sera, Nobuyuki; Horikawa, Kazumi

    2015-03-01

    Recently, there has been a marked increase in the number of reports of fluoroquinolone-resistant Campylobacter jejuni and Campylobacter coli. The aim of this study was to evaluate the prevalence of antimicrobial resistance and its genetic determinants in Campylobacter species isolated from meat and human subjects in Fukuoka Prefecture, Japan. Between 2011 and 2013, 55 and 64 isolates were collected from meat (chicken meat and beef liver) and humans, respectively, in this prefecture. Antimicrobial susceptibility tests were conducted using the agar dilution method in accordance with the Clinical and Laboratory Standards Institute guidelines, using the following 11 antimicrobial agents : cephalexin, cefoxitin, nalidixic acid, ciprofloxacin, levofloxacin, tetracycline, minocycline, ampicillin, streptomycin, kanamycin and erythromycin. The susceptibility rates of the isolates to three quinolones (nalidixic acid, ciprofloxacin, levofloxacin) were 43.7%, 41.2%, 40.3%, respectively. All the isolates were multidrug resistant. Whereas 46.9%-51.6% of the human isolates were resistant to one or more of the quinolones, only 32.7%-34.5% of the meat isolates were resistant to one or more of the drugs. DNA sequencing showed that of the 50 quinolone resistant isolates 44 had position 86 isoleucine (Ile) substituted for threonine (Thr) in the GyrA protein (Thr86Ile). This amino acid substitution resulted from ACA to ATA and ACT to ATT mutations of codon 86 in C. jejuni and C. coli, respectively. Furthermore, two of the four C. jejuni isolates lacking the Thr86Ile mutation had combined Ser22Gly-Asn203Ser substitutions, while the remaining two isolates had combined Ser22Gly-Asn203Ser-Ala 206Val substitutions. These four isolates also had cmeABC sequences that differed from the quinolone sensitive C. jejuni ATCC33560(T) strain. In conclusion, C. jejuni and C. coli have relatively high quinolone resistance, and are resistant to other antibiotics. The new combination of amino acid

  20. [Antimicrobial Susceptibility and Resistance Mutations in Campylobacter jejuni and C. coli Isolates from Human and Meat Sources].

    PubMed

    Oishi, Akira; Murakami, Koichi; Etoh, Yoshiki; Sera, Nobuyuki; Horikawa, Kazumi

    2015-03-01

    Recently, there has been a marked increase in the number of reports of fluoroquinolone-resistant Campylobacter jejuni and Campylobacter coli. The aim of this study was to evaluate the prevalence of antimicrobial resistance and its genetic determinants in Campylobacter species isolated from meat and human subjects in Fukuoka Prefecture, Japan. Between 2011 and 2013, 55 and 64 isolates were collected from meat (chicken meat and beef liver) and humans, respectively, in this prefecture. Antimicrobial susceptibility tests were conducted using the agar dilution method in accordance with the Clinical and Laboratory Standards Institute guidelines, using the following 11 antimicrobial agents : cephalexin, cefoxitin, nalidixic acid, ciprofloxacin, levofloxacin, tetracycline, minocycline, ampicillin, streptomycin, kanamycin and erythromycin. The susceptibility rates of the isolates to three quinolones (nalidixic acid, ciprofloxacin, levofloxacin) were 43.7%, 41.2%, 40.3%, respectively. All the isolates were multidrug resistant. Whereas 46.9%-51.6% of the human isolates were resistant to one or more of the quinolones, only 32.7%-34.5% of the meat isolates were resistant to one or more of the drugs. DNA sequencing showed that of the 50 quinolone resistant isolates 44 had position 86 isoleucine (Ile) substituted for threonine (Thr) in the GyrA protein (Thr86Ile). This amino acid substitution resulted from ACA to ATA and ACT to ATT mutations of codon 86 in C. jejuni and C. coli, respectively. Furthermore, two of the four C. jejuni isolates lacking the Thr86Ile mutation had combined Ser22Gly-Asn203Ser substitutions, while the remaining two isolates had combined Ser22Gly-Asn203Ser-Ala 206Val substitutions. These four isolates also had cmeABC sequences that differed from the quinolone sensitive C. jejuni ATCC33560(T) strain. In conclusion, C. jejuni and C. coli have relatively high quinolone resistance, and are resistant to other antibiotics. The new combination of amino acid

  1. The cinnamon-oil ingredient trans-cinnamaldehyde fails to target Campylobacter jejuni strain KC 40 in the broiler chicken cecum despite marked in vitro activity.

    PubMed

    Hermans, David; Martel, An; van Deun, Kim; van Immerseel, Filip; Heyndrickx, Marc; Haesebrouck, Freddy; Pasmans, Frank

    2011-10-01

    Campylobacter jejuni is the most common bacterial cause of diarrheal disease in humans worldwide, with poultry products being a major source. Therefore, strategies to decrease Campylobacter colonization during primary production might aid in reducing the number of human campylobacteriosis cases. Several plant-derived compounds have been reported to possess anti-Campylobacter properties in vitro, so they could be promising candidates to reduce Campylobacter colonization in broiler chickens. To test this hypothesis, selected plant-derived antimicrobials (caffeic, gallic, protocatechuic, and vanillic acids, epigallocatechin gallate, trans-cinnamaldehyde, and thymol) were screened for anti-Campylobacter activity by determining MICs and setting up time-kill curves for C. jejuni strain KC 40. These experiments revealed marked antibacterial activity, especially for the cinnamon oil ingredient trans-cinnamaldehyde (CIN). This compound was tested in a broiler chick seeder model; it was added to the feed in coated form at an effective concentration of 0.3 % from day-of-hatch for the entire 22-day duration of the experiment. At 14 days of age, one-third of the birds were inoculated with C. jejuni strain KC 40 and served as seeders. CIN was not able to reduce cecal Campylobacter colonization in this model, which was confirmed in a cecal loop experiment. Despite CIN concentrations much higher than the MIC, C. jejuni numbers were not reduced compared with those in nontreated ceca at 2 and 24 h after injection. In conclusion, this study shows a marked discrepancy between in vitro and in vivo activity of CIN against C. jejuni strain KC 40. PMID:22004822

  2. Effects of Sequential Campylobacter jejuni 81-176 Lipooligosaccharide Core Truncations on Biofilm Formation, Stress Survival, and Pathogenesis▿ †

    PubMed Central

    Naito, Mizue; Frirdich, Emilisa; Fields, Joshua A.; Pryjma, Mark; Li, Jianjun; Cameron, Andrew; Gilbert, Michel; Thompson, Stuart A.; Gaynor, Erin C.

    2010-01-01

    Campylobacter jejuni is a highly prevalent human pathogen for which pathogenic and stress survival strategies remain relatively poorly understood. We previously found that a C. jejuni strain 81-176 mutant defective for key virulence and stress survival attributes was also hyper-biofilm and hyperreactive to the UV fluorescent dye calcofluor white (CFW). We hypothesized that screening for CFW hyperreactive mutants would identify additional genes required for C. jejuni pathogenesis properties. Surprisingly, two such mutants harbored lesions in lipooligosaccharide (LOS) genes (waaF and lgtF), indicating a complete loss of the LOS outer core region. We utilized this as an opportunity to explore the role of each LOS core-specific moiety in the pathogenesis and stress survival of this strain and thus also constructed ΔgalT and ΔcstII mutants with more minor LOS truncations. Interestingly, we found that mutants lacking the LOS outer core (ΔwaaF and ΔlgtF but not ΔgalT or ΔcstII mutants) exhibited enhanced biofilm formation. The presence of the complete outer core was also necessary for resistance to complement-mediated killing. In contrast, any LOS truncation, even that of the terminal sialic acid (ΔcstII), resulted in diminished resistance to polymyxin B. The cathelicidin LL-37 was found to be active against C. jejuni, with the LOS mutants exhibiting modest but tiled alterations in LL-37 sensitivity. The ΔwaaF mutant but not the other LOS mutant strains also exhibited a defect in intraepithelial cell survival, an aspect of C. jejuni pathogenesis that has only recently begun to be clarified. Finally, using a mouse competition model, we now provide the first direct evidence for the importance of the C. jejuni LOS in host colonization. Collectively, this study has uncovered novel roles for the C. jejuni LOS, highlights the dynamic nature of the C. jejuni cell envelope, and provides insight into the contribution of specific LOS core moieties to stress survival and

  3. Comparative proteomics and glycoproteomics reveal increased N-linked glycosylation and relaxed sequon specificity in Campylobacter jejuni NCTC11168 O.

    PubMed

    Scott, Nichollas E; Marzook, N Bishara; Cain, Joel A; Solis, Nestor; Thaysen-Andersen, Morten; Djordjevic, Steven P; Packer, Nicolle H; Larsen, Martin R; Cordwell, Stuart J

    2014-11-01

    Campylobacter jejuni is a major cause of bacterial gastroenteritis. C. jejuni encodes a protein glycosylation (Pgl) locus responsible for the N-glycosylation of membrane-associated proteins. We examined two variants of the genome sequenced strain NCTC11168: O, a representative of the original clinical isolate, and GS, a laboratory-adapted relative of O. Comparative proteomics by iTRAQ and two-dimensional liquid chromatography coupled to tandem mass spectrometry (2D-LC-MS/MS) allowed the confident identification of 1214 proteins (73.9% of the predicted C. jejuni proteome), of which 187 were present at statistically significant altered levels of abundance between variants. Proteins associated with the O variant included adhesins (CadF and FlpA), proteases, capsule biosynthesis, and cell shape determinants as well as six proteins encoded by the Pgl system, including the PglK flippase and PglB oligosaccharyltransferase. Lectin blotting highlighted specific glycoproteins more abundant in NCTC11168 O, whereas others remained unaltered. Hydrophilic interaction liquid chromatography (HILIC) and LC-MS/MS identified 30 completely novel glycosites from 15 proteins. A novel glycopeptide from a 14 kDa membrane protein (Cj0455c) was identified that did not contain the C. jejuni N-linked sequon D/E-X-N-X-S/T (X ≠ Pro) but that instead contained a sequon with leucine at the -2 position. Occupied atypical sequons were also observed in Cj0958c (OxaA; Gln at the -2 position) and Cj0152c (Ala at the +2 position). The relative O and GS abundances of 30 glycopeptides were determined by label-free quantitation, which revealed a >100-fold increase in the atypical glycopeptide from Cj0455c in isolate O. Our data provide further evidence for the importance of the Pgl system in C. jejuni.

  4. Prediction of CTL epitope, in silico modeling and functional analysis of cytolethal distending toxin (CDT) protein of Campylobacter jejuni

    PubMed Central

    2014-01-01

    Background Campylobacter jejuni is a potent bacterial pathogen culpable for diarrheal disease called campylobacteriosis. It is realized as a major health issue attributable to unavailability of appropriate vaccines and clinical treatment options. As other pathogens, C. jejuni entails host cellular components of an infected individual to disseminate this disease. These host–pathogen interfaces during C. jejuni infection are complex, vibrant and involved in the nicking of host cell environment, enzymes and pathways. Existing therapies are trusted only on a much smaller number of drugs, most of them are insufficient because of their severe host toxicity or drug-resistance phenomena. To find out remedial alternatives, the identification of new biotargets is highly anticipated. Understanding the molecules involved in pathogenesis has the potential to yield new and exciting strategies for therapeutic intervention. In this direction, advances in bioinformatics have opened up new possibilities for the rapid measurement of global changes during infection and this could be exploited to understand the molecular interactions involved in campylobacteriosis. Methods In this study, homology modeling, epitope prediction and identification of ligand binding sites has been explored. Further attempt to generate strapping 3D model of cytolethal distending toxin protein from C. jejuni have been described for the first time. Results CDT protein isolated from C. jejuni was analyzed using various bioinformatics and immuno-informatics tools including sequence and structure tools. A total of fifty five antigenic determinants were predicted and prediction results of CTL epitopes revealed that five MHC ligand are found in CDT. The three potential pocket binding site are found in the sequence that can be useful for drug designing. Conclusions This model, we hope, will be of help in designing and predicting novel CDT inhibitors and vaccine candidates. PMID:24552167

  5. Campylobacter jejuni Biofilms Up-Regulated in the Absence of the Stringent Response Utilize a Calcofluor White-Reactive Polysaccharide▿

    PubMed Central

    McLennan, Meghan K.; Ringoir, Danielle D.; Frirdich, Emilisa; Svensson, Sarah L.; Wells, Derek H.; Jarrell, Harold; Szymanski, Christine M.; Gaynor, Erin C.

    2008-01-01

    The enteric pathogen Campylobacter jejuni is a highly prevalent yet fastidious bacterium. Biofilms and surface polysaccharides participate in stress survival, transmission, and virulence in C. jejuni; thus, the identification and characterization of novel genes involved in each process have important implications for pathogenesis. We found that C. jejuni reacts with calcofluor white (CFW), indicating the presence of surface polysaccharides harboring β1-3 and/or β1-4 linkages. CFW reactivity increased with extended growth, under 42°C anaerobic conditions, and in a ΔspoT mutant defective for the stringent response (SR). Conversely, two newly isolated dim mutants exhibited diminished CFW reactivity as well as growth and serum sensitivity differences from the wild type. Genetic, biochemical, and nuclear magnetic resonance analyses suggested that differences in CFW reactivity between wild-type and ΔspoT and dim mutant strains were independent of well-characterized lipooligosaccharides, capsular polysaccharides, and N-linked polysaccharides. Targeted deletion of carB downstream of the dim13 mutation also resulted in CFW hyporeactivity, implicating a possible role for carbamoylphosphate synthase in the biosynthesis of this polysaccharide. Correlations between biofilm formation and production of the CFW-reactive polymer were demonstrated by crystal violet staining, scanning electron microscopy, and confocal microscopy, with the C. jejuni ΔspoT mutant being the first SR mutant in any bacterial species identified as up-regulating biofilms. Together, these results provide new insight into genes and processes important for biofilm formation and polysaccharide production in C. jejuni. PMID:17993532

  6. Dynamics of Dual Infection with Campylobacter jejuni Strains in Chickens Reveals Distinct Strain-to-Strain Variation in Infection Ecology

    PubMed Central

    Wigley, Paul; Humphrey, Suzanne; Kemmett, Kirsty; Lacharme-Lora, Lizeth; Humphrey, Tom; Williams, Nicola

    2014-01-01

    Although multiple genotypes of Campylobacter jejuni may be isolated from the same commercial broiler flock, little is known about the infection dynamics of different genotypes within individuals or their colonization sites within the gut. Single experimental infections with C. jejuni M1 (sequence type 137, clonal complex 45) and C. jejuni 13126 (sequence type 21, clonal complex 21) revealed that 13126 colonized the ceca at significantly higher levels. The dissemination and colonization sites of the two C. jejuni strains then were examined in an experimental broiler flock. Two 33-day-old broiler chickens were infected with M1 and two with 13126, and 15 birds were left unchallenged. Cloacal swabs were taken postinfection to determine the colonization and shedding of each strain. By 2 days postinfection (dpi), 8/19 birds were shedding M1 whereas none were shedding 13126. At 8 dpi, all birds were shedding both strains. At 18 dpi, liver and cecal levels of each isolate were quantified, while in 10 birds they also were quantified at nine sites throughout the gastrointestinal (GI) tract. 13126 was found throughout the GI tract, while M1 was largely restricted to the ceca and colon. The livers of 7/19 birds were culture positive for 13126 only. These data show that 13126 has a distinctly different infection biology than strain M1. It showed slower colonization of the lower GI tract but was more invasive and able to colonize at a high level throughout the GI tract. The finding that C. jejuni strains have markedly different infection ecologies within the chicken has implications for control in the poultry industry and suggests that the contamination risk of edible tissues is dependent on the isolate involved. PMID:25107966

  7. Multilocus sequence typing and FlaA sequencing reveal the genetic stability of Campylobacter jejuni enrichment during coculture with Acanthamoeba polyphaga.

    PubMed

    Griekspoor, Petra; Olofsson, Jenny; Axelsson-Olsson, Diana; Waldenström, Jonas; Olsen, Björn

    2013-04-01

    Low concentrations of Campylobacter jejuni cells in environmental samples make them difficult to study with conventional culture methods. Here, we show that enrichment by amoeba cocultures works well with low-concentration samples and that this method can be combined with molecular techniques without loss of genetic specificity. PMID:23377942

  8. A poultry-intestinal isolate of Campylobacter jejuni produces a bacteriocin (CUV-3) active against a range of Gram positive bacterial pathogens including Clostridium perfringens

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A newly isolated bacteriocin, CUV-3, produced by a poultry cecal isolate of Campylobacter jejuni strain CUV-3 had inhibitory activity against several Gram positive bacteria including Clostridium perfringens (38 strains), Staphylococcus aureus, Staph.epidermidis and Listeria monocytogenes. The pept...

  9. Analysis of the Pan Genome of Campylobacter jejuni Isolates Recovered from Poultry by Pulse-Field Gel Electrophoreis, MLST and rep-PCR Reveals Different Discriminatory Capabilities

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Campylobacter jejuni is one of the leading bacterial causes of food-borne illness in the United States. Molecular genotyping methods are often used in food safety for identifying sources of infection and pathways of transmission. Moreover, the identification of genetically related strains (i.e., cla...

  10. Crystal Structure of the Transcriptional Regulator CmeR From Campylobacter Jejuni

    SciTech Connect

    Gu, R.; Su, C.-C.; Shi, F.; McDermott, G.; Zhang, Q.; Yu, E.W.

    2009-06-01

    The CmeABC multidrug efflux pump, which belongs to the resistance-nodulation-division (RND) family, recognizes and extrudes a broad range of antimicrobial agents and is essential for Campylobacter jejuni colonization of the animal intestinal tract by mediating the efflux of bile acids. The expression of CmeABC is controlled by the transcriptional regulator CmeR, whose open reading frame is located immediately upstream of the cmeABC operon. To understand the structural basis of CmeR regulation, we have determined the crystal structure of CmeR to 2.2 {angstrom} resolution, revealing a dimeric two-domain molecule with an entirely helical architecture similar to members of the TetR family of transcriptional regulators. Unlike the rest of the TetR regulators, CmeR has a large center-to-center distance (54 {angstrom}) between two N termini of the dimer, and a large flexible ligand-binding pocket in the C-terminal domain. Each monomer forms a 20 {angstrom} long tunnel-like cavity in the ligand-binding domain of CmeR and is occupied by a fortuitous ligand that is identified as glycerol. The binding of glycerol to CmeR induces a conformational state that is incompatible with target DNA. As glycerol has a chemical structure similar to that of potential ligands of CmeR, the structure obtained mimics the induced form of CmeR. These findings reveal novel structural features of a TetR family regulator, and provide new insight into the mechanisms of ligand binding and CmeR regulation.

  11. Campylobacter jejuni colonization promotes the translocation of Escherichia coli to extra-intestinal organs and disturbs the short-chain fatty acids profiles in the chicken gut.

    PubMed

    Awad, W A; Dublecz, F; Hess, C; Dublecz, K; Khayal, B; Aschenbach, J R; Hess, M

    2016-10-01

    For a long time Campylobacter was only considered as a commensal microorganism in avian hosts restricted to the ceca, without any pathogenic features. The precise reasons for the symptomless chicken carriers are still unknown, but investigations of the gastrointestinal ecology of broiler chickens may improve our understanding of the microbial interactions with the host. Therefore, the current studies were conducted to investigate the effects of Campylobacter jejuni colonization on Escherichia coli translocation and on the metabolic end products (short-chain fatty acids, SCFAs). Following oral infection of 14 day old broiler chickens with 1 × 10(8) CFU of Campylobacter jejuni NCTC 12744 in two independent animal trials, it was found that C. jejuni heavily colonized the intestine and disseminate to extra-intestinal organs. Moreover, in both animal trials, the findings revealed that C. jejuni promoted the translocation of E. coli with a higher number encountered in the spleen and liver at 14 days post infection (dpi). In addition, Campylobacter affected the microbial fermentation in the gastrointestinal tract of broilers by reducing the amount of propionate, isovalerate, and isobutyrate in the cecal digesta of the infected birds at 2 dpi and, at 7 and 14 dpi, butyrate, isobutyrate, and isovalerate were also decreased. However, in the jejunum, the C. jejuni infection lowered only butyrate concentrations at 14 dpi. These data indicated that C. jejuni may utilize SCFAs as carbon sources to promote its colonization in the chicken gut, suggesting that Campylobacter cannot only alter gut colonization dynamics but might also influence physiological processes due to altered microbial metabolite profiles.Finally, the results demonstrated that C. jejuni can cross the intestinal epithelial barrier and facilitates the translocation of Campylobacter itself as well as of other enteric microorganisms such as E. coli to extra-intestinal organs of infected birds. Altogether, our

  12. The role of Campylobacter jejuni cytolethal distending toxin in gastroenteritis: toxin detection, antibody production, and clinical outcome

    SciTech Connect

    Boisen, Nadia

    2011-01-01

    The role of Campylobacter jejuni cytolethal distending toxin (CDT) on clinical outcome after gastroenteritis was investigated. Clinical data, blood serum samples, and Campylobacter spp. isolated, from each of 30 patients were collected over a period of 6 months. The CDT encoding genes, cdtABC, characterized by PCR, revealed that all but one of the C. jejuni strains had the wild-type sequence. Sequencing of cdtABC from this strain showed two major deletions. From all of the strains, CDT titers were determined, and toxin neutralizing antibodies were documented using an in vitro assay. Three of the thirty clinical isolates, including the one with the mutant cdtABC coding genes, did not have a detectable CDT activity. Analyzing the relationship between CDT titer, serum neutralization of CDT, and the clinical outcome showed that campylobacteriosis caused by CDT-negative strains was clinically indistinguishable from that of patients infected with an isolate that produced high levels of CDT. These results suggest that CDT does not solely determine severity of infection and clinical outcome.

  13. A foodborne outbreak of Campylobacter jejuni (O:33) infection associated with tuna salad: a rare strain in an unusual vehicle.

    PubMed Central

    Roels, T. H.; Wickus, B.; Bostrom, H. H.; Kazmierczak, J. J.; Nicholson, M. A.; Kurzynski, T. A.; Davis, J. P.

    1998-01-01

    We report a foodborne outbreak of Campylobacter jejuni infection in a summer camp. Outbreak-related cases occurred in 79 persons including 3 secondary cases in campers. Campylobacter jejuni was isolated from stool specimens from 16 of 21 patients who submitted a sample; 13 viable isolates were serotyped and all were serotype O:33 (somatic O scheme) or HL:18 (heat-labile scheme), and biotype III (Lior scheme). This serotype is widely distributed geographically but rarely isolated from humans. Samples of water from the wells supplying the camp were negative for faecal coliforms, and raw milk had not been served in the camp. A matched (1:1) case-control study identified tuna salad served for lunch on 19 July as the likely food item associated with illness (matched odds ratio=22; 95% confidence intervals (CI)=3.6-908). Swimming in the camp pool and other recreational water use in area lakes by the campers were not statistically associated with illness. The precise mechanism of introduction of the organism into the tuna salad remains unknown; contamination most likely occurred through cross-contamination with another food product, the hands of a food handler, or a work surface. Several deficiencies in the operation of the camp kitchen were identified. In Wisconsin, kitchens of such camps are subject to different inspection rules than restaurants. Camp staff, administrators, counselors, food managers, and infirmary staff, should fulfil important roles in their respective areas to prevent future outbreaks. PMID:9825778

  14. Polyphosphate kinases modulate Campylobacter jejuni outer membrane constituents and alter its capacity to invade and survive in intestinal epithelial cells in vitro

    PubMed Central

    Pina-Mimbela, Ruby; Madrid, Jesús Arcos; Kumar, Anand; Torrelles, Jordi B; Rajashekara, Gireesh

    2015-01-01

    Campylobacter jejuni is the most prevalent cause of bacterial gastroenteritis worldwide. Polyphosphate kinases 1 and 2 (PPK1 and PPK2) regulate several cellular processes, including the biosynthesis of the bacterial cell wall. Despite their importance, whether PPK1 and PPK2 modulate the composition of C. jejuni outer membrane constituents (OMCs) and consequently impact its interaction with host cells remains unknown. Our comparative analysis between C. jejuni wild type, Δppk1, and Δppk2 strains showed qualitative and quantitative differences in the total OMC composition among these strains. Importantly, these OMC variations observed on the C. jejuni polyphosphate kinase mutants are directly related to their capacity to invade, survive, and alter the immune response of intestinal epithelial cells in vitro. Specifically, sub-fractionation of the C. jejuni OMC indicated that OMC proteins are uniquely associated with bacterial invasion, whereas C. jejuni OMC proteins, lipids, and li