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Sample records for campylobacter jejuni isolated

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

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    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. PMID:24210812

  2. Antimicrobial activities of isothiocyanates against Campylobacter jejuni isolates

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    Virginie eDufour

    2012-04-01

    Full Text Available Food-borne human infection with Campylobacter jejuni is a medical concern in both industrialized and developing countries. Efficient eradication of C. jejuni reservoirs within live animals and processed foods is limited by the development of antimicrobial resistances and by practical problems related to the use of conventional antibiotics in food processes.We have investigated the bacteriostatic and bactericidal activities of two phytochemicals, allyl-isothiocyanate (AITC and benzyl-isothiocyanate (BITC, against 24 C. jejuni isolates from chicken feces, human infections and contaminated foods, as well as two reference strains NCTC11168 and 81-176.Both AITC and BITC displayed a potent antibacterial activity against C. jejuni. BITC showed a higher overall antibacterial effect (MIC of 2.5 to 5 g mL-1 compared to AITC (MIC of 50 to 200 g mL-1. Interestingly, the 24 C. jejuni isolates could be classified in 3 groups according to their sensitivity levels to both compounds, suggesting that AITC and BITC shared identical activity mechanisms and consequently faced similar resistance processes in bacterial cells.The sensitivity levels of C. jejuni strains against isothiocyanates were neither correlated with the presence of a GGT (-Glutamyl Transpeptidase encoding gene in the genome nor with the origin of the biological sample. However the ggt mutant of C. jejuni 81-176 displayed a decreased survival rate compared to WT when exposed to ITC.

  3. Isolation and characterization of Campylobacter jejuni subsp jejuni from macaroni penguins (Eudyptes chrysolophus) in the subantarctic region

    DEFF Research Database (Denmark)

    Broman, T.; Bergstrom, S.; On, Stephen L.W.;

    2000-01-01

    On Bird Island, South Georgia, albatrosses (n = 140), penguins (n = 100), and fur seals (n = 206) were sampled for Campylobacter jejuni. C. jejuni subsp. jejuni was recovered from three macaroni penguins (Eudyptes chrysolophus). These isolates, the first reported for the subantarctic region, showed...

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

    DEFF Research Database (Denmark)

    Sørensen, Martine C. Holst; Gencay, Yilmaz Emre; Birk, Tina;

    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...

  5. Campylobacter jejuni organism (image)

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    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 ...

  6. Genotypes and antibiotic resistances of Campylobacter jejuni and Campylobacter coli isolates from domestic and travel-associated human cases

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    Niederer, Lilian; Kuhnert, Peter; Egger, Ralph; Büttner, Sabina; Hächler, Herbert; Korczak, Bozena M.

    2012-01-01

    Multilocus sequence typing (MLST) extended with flaB typing of 425 Campylobacter jejuni isolates and 42 Campylobacter coli isolates revealed quite a low overlap between human isolates from travel-associated and domestic cases in Switzerland. Men were more frequently affected by Campylobacter than women, but strains from women and, overall, from travel-associated cases showed mutations conferring quinolone resistance more frequently than strains from men and domestic cases, respectively.

  7. Isolation and detection of Campylobacter jejuni from chicken fecal samples by immunomagnetic separation–PCR

    DEFF Research Database (Denmark)

    Le Ly, Tram Thuy; Cao, Cuong; Høgberg, Jonas;

    2012-01-01

    Campylobacter jejuni (C. jejuni) is one of the leading causes of bacterial food-borne disease worldwide. The presence of Campylobacter in chicken feces poses a high risk for contamination of chicken meat and for Campylobacter infections in human. Detection of this bacterium in chicken fecal...... of the assay was assured by two selection steps: 1) Dynabeads®M-270 Amine microbeads (2.8 μm in diameter) coated with C. jejuni monoclonal antibodies were used as the primary selection to isolate bacteria from fecal samples. 2) A PCR assay amplifying the Hippuricase gene was performed as the specific selection...

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

    OpenAIRE

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

    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 resis...

  9. Molecular typing of Campylobacter jejuni isolates involved in a neonatal outbreak indicates nosocomial transmission

    DEFF Research Database (Denmark)

    Llovo, J.; Mateo, E.; Munoz, A.;

    2003-01-01

    Genotypic typing by restriction fragment length polymorphism and pulsed-field gel electrophoresis showed that two neonates in a neonatal ward were infected with the same Campylobacter jejuni strain. Isolates from the mother and brother of the index patient were identical to each other but distinc...... from the neonatal type. Genotyping results therefore suggested that the neonatal C. jejuni infection was nosocomial in origin.......Genotypic typing by restriction fragment length polymorphism and pulsed-field gel electrophoresis showed that two neonates in a neonatal ward were infected with the same Campylobacter jejuni strain. Isolates from the mother and brother of the index patient were identical to each other but distinct...

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

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    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.

  11. Arsenic Resistance and Prevalence of Arsenic Resistance Genes in Campylobacter jejuni and Campylobacter coli Isolated from Retail Meats

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    Mohamed K. Fakhr

    2013-08-01

    Full Text Available Studies that investigate arsenic resistance in the foodborne bacterium Campylobacter are limited. A total of 552 Campylobacter isolates (281 Campylobacter jejuni and 271 Campylobacter coli isolated from retail meat samples were subjected to arsenic resistance profiling using the following arsenic compounds: arsanilic acid (4–2,048 μg/mL, roxarsone (4–2048 μg/mL, arsenate (16–8,192 μg/mL and arsenite (4–2,048 μg/mL. A total of 223 of these isolates (114 Campylobacter jejuni and 109 Campylobacter coli were further analyzed for the presence of five arsenic resistance genes (arsP, arsR, arsC, acr3, and arsB by PCR. Most of the 552 Campylobacter isolates were able to survive at higher concentrations of arsanilic acid (512–2,048 μg/mL, roxarsone (512–2,048 μg/mL, and arsenate (128–1,024 μg/mL, but at lower concentrations for arsenite (4–16 μg/mL. Ninety seven percent of the isolates tested by PCR showed the presence of arsP and arsR genes. While 95% of the Campylobacter coli isolates contained a larger arsenic resistance operon that has all of the four genes (arsP, arsR, arsC and acr3, 85% of the Campylobacter jejuni isolates carried the short operon (arsP, and arsR. The presence of arsC and acr3 did not significantly increase arsenic resistance with the exception of conferring resistance to higher concentrations of arsenate to some Campylobacter isolates. arsB was prevalent in 98% of the tested Campylobacter jejuni isolates, regardless of the presence or absence of arsC and acr3, but was completely absent in Campylobacter coli. To our knowledge, this is the first study to determine arsenic resistance and the prevalence of arsenic resistance genes in such a large number of Campylobacter isolates.

  12. An improved culture method for selective isolation of Campylobacter jejuni from wastewater

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    Jinyong Kim

    2016-08-01

    Full Text Available 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 (qRT-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. In particular, rifampicin supplementation and an increased culture temperature (i.e., 42°C had a decisive effect on the selective enrichment of C. jejuni from wastewater. 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.

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

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    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.

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

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    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

  15. Antibiotic resistance profile and RAPD analysis of Campylobacter jejuni isolated from vegetables farms and retail markets

    Institute of Scientific and Technical Information of China (English)

    John Yew Huat Tang; Mohd Ikhsan Khalid; Syazana Aimi; Che Abdullah Abu-Bakar; Son Radu

    2016-01-01

    Objective: To investigate antibiotic resistance profile and characterize Campylobacter jejuni (C. jejuni) isolates using random amplified polymorphic DNA (RAPD) analysis. Methods: Ninety eight C. jejuni isolates from farms and retail outlets were screened against 10 antibiotics commonly used clinically and agriculturally by using disk diffusion method. RAPD analysis was done to characterize 98 C. jejuni isolates. Results: Fifty-one percent of the isolates had multiple antibiotic resistance index 0.2 and below. This indicated that the isolates in the vegetables were not from the high risk environment or extensive farming practices. C. jejuni isolates found resistant towards penicillin G (93%), vancomycin (86%), ampicillin (35%), erythromycin (28%), genta-mycin (4%), amikacin (3%), enrofloxacin (1%), norfloxacin (1%) and no resistance to-wards ciprofloxacin. RAPD clustering analysis showed that the contamination of C. jejuni in vegetables was likely due to cross contamination at retail markets. Conclusions: C. jejuni contamination in vegetables at retail markets was due to cross contamination. Current finding proved that C. jejuni in small scale vegetables production was less expose towards antibiotic abuse.

  16. Prevalence of virulence genes and cytolethal distending toxin production in Campylobacter jejuni isolates from diarrheal patients in Bangladesh

    NARCIS (Netherlands)

    Talukder, Kaisar A.; Aslam, Mohammad; Islam, Zhahirul; Azmi, Ishrat J.; Dutta, Dilip K.; Hossain, Sabir; Nur-E-Kamal, Alam; Nair, Gopinath B.; Cravioto, Alejandro; Sack, David A.; Endtz, Hubert P.

    2008-01-01

    From 300 stool samples, 58 Campylobacter strains were isolated by standard microbiological and biochemical methods. Of these, 40 strains were identified as Campylobacter jejuni and 5 as Campylobacter coli. The presence of flaA (100%), cadF (100%), racR (100%), dnaJ (100%), pldA (100%), ciaB (95%), v

  17. Evaluation of phenotypic and genotypic methods for subtyping Campylobacter jejuni isolates from humans, poultry, and cattle

    DEFF Research Database (Denmark)

    Nielsen, Eva Møller; Engberg, J.; Fussing, V.;

    2000-01-01

    Six methods for subtyping of Campylobacter jejuni were compared and evaluated with a collection of 90 isolates from poultry, cattle, and sporadic human clinical cases as well as from a waterborne outbreak. The applied methods were Penner heat-stable serotyping; automated ribotyping (Ribo...

  18. Susceptibilities of Campylobacter jejuni Isolates from Germany to Ciprofloxacin, Moxifloxacin, Erythromycin, Clindamycin, and Tetracycline

    OpenAIRE

    Wagner, Jutta; Jabbusch, Miriam; Eisenblätter, Martin; Hahn, Helmut; Wendt, Constanze; Ignatius, Ralf

    2003-01-01

    To elucidate Campylobacter jejuni resistance to antibiotics in Germany, MICs of ciprofloxacin, moxifloxacin, erythromycin, clindamycin, and tetracycline were determined (using agar dilution) for 144 clinical isolates. The data indicate a considerable ciprofloxacin resistance (45.1%) without a clonal relationship of the strains and a greater in vitro activity of moxifloxacin, erythromycin, and clindamycin.

  19. Antimicrobial resistance among Campylobacter jejuni isolated from raw poultry meat at retail level in Denmark

    DEFF Research Database (Denmark)

    Andersen, S. R.; Saadbye, P.; Shukri, Naseer Mahmoud;

    2006-01-01

    Campylobacter jejuni isolated from raw poultry meat collected at retail shops in Denmark in the period 1996-2003 were tested for susceptibility to seven antimicrobial agents. The food samples consisted of raw chicken meat and other raw poultry meat of domestic or imported origin. The highest levels...... of resistance among C. jejuni were observed for tetracycline, nalidixic acid and ciprofloxacin, whereas macrolide resistance was rarely detected. C. jejuni originating from other poultry meat (mainly duck and turkey meat) exhibited the highest occurrences of antimicrobial resistance monitored; approximately one...... third of the isolates were tetracycline resistant (N=100). Among chicken meat isolates, the occurrence of tetracycline resistance was significantly higher (P chicken meat (N=88) than in C. jejuni from Danish chicken meat (N=367). The same tendency was observed...

  20. Multilocus sequence types of Finnish bovine Campylobacter jejuni isolates and their attribution to human infections

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    Corander Jukka

    2010-07-01

    Full Text Available Abstract Background Campylobacter jejuni is the most common bacterial cause of human gastroenteritis worldwide. Due to the sporadic nature of infection, sources often remain unknown. Multilocus sequence typing (MLST has been successfully applied to population genetics of Campylobacter jejuni and mathematical modelling can be applied to the sequence data. Here, we analysed the population structure of a total of 250 Finnish C. jejuni isolates from bovines, poultry meat and humans collected in 2003 using a combination of Bayesian clustering (BAPS software and phylogenetic analysis. Results In the first phase we analysed sequence types (STs of 102 Finnish bovine C. jejuni isolates by MLST and found a high diversity totalling 50 STs of which nearly half were novel. In the second phase we included MLST data from domestic human isolates as well as poultry C. jejuni isolates from the same time period. Between the human and bovine isolates we found an overlap of 72.2%, while 69% of the human isolates were overlapping with the chicken isolates. In the BAPS analysis 44.3% of the human isolates were found in bovine-associated BAPS clusters and 45.4% of the human isolates were found in the poultry-associated BAPS cluster. BAPS reflected the phylogeny of our data very well. Conclusions These findings suggest that bovines and poultry were equally important as reservoirs for human C. jejuni infections in Finland in 2003. Our results differ from those obtained in other countries where poultry has been identified as the most important source for human infections. The low prevalence of C. jejuni in poultry flocks in Finland could explain the lower attribution of human infection to poultry. Of the human isolates 10.3% were found in clusters not associated with any host which warrants further investigation, with particular focus on waterborne transmission routes and companion animals.

  1. Characterization of Campylobacter jejuni and Campylobacter coli strains isolated in the region of Niš, Serbia

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    Miljković-Selimović Biljana

    2010-01-01

    Full Text Available Introduction. Campylobacter jejuni and Campylobacter coli represent one of the main causes of bacterial diarrhoea in humans. Although the disease is usually mild and self-limiting, severe chronic sequelae may occur, such as reactive arthritis, Guillain-Barré and Miller Fisher syndromes. Serotyping is used as an epidemiological marker, while post-infective polyneuropathies are associated with several O serotypes. Objective. Strains of C. jejuni and C. coli were serotyped based on heat stable (HS and heat labile (HL antigens, as well as biotypes to determine strain diversity. Methods. Campylobacter spp. was isolated using selective blood media with antibiotics. Differentiation to the species level was done by a combination of biotyping tests and by a PCR-based RFLP test. The isolates were characterised by Penner and Lior serotyping methods. Results. The serotypes showed diversity without predominant serotypes. 24 HS serotypes were detected among 29 C. jejuni strains, and seven serotypes among nine C. coli strains. HL serotyping method successfully typed 62.5% of strains. Among 16 C. jejuni strains 14 serotypes were detected, and three among four C. coli strains. A C. jejuni strain associated with a patient with Guillain-Barré syndrome was typed as biotype II, O:19. Conclusion. The biotyping and serotyping results have indicated that C. jejuni and C. coli strains in the region of Niš, Serbia are diverse and could be probably of unrelated sources of origin or reservoirs. The strain associated with the Guillain-Barré syndrome patient was serotype O:19, one of the most common in this post-infective complication.

  2. Primary isolation strain determines both phage type and receptors recognised by Campylobacter jejuni bacteriophages.

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    Martine C Holst Sørensen

    Full Text Available 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.

  3. Antibiotic susceptibility profiling and virulence potential ofCampylobacter jejuni isolates from different sources in Pakistan

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

    2015-01-01

    Objective:To determine antibiotic resistance patterns and virulence potential ofCampylobacter jejuni (C. jejuni) isolates from clinical human diarrheal infections, cattle and healthy broilers. Methods:Antibiotic sensitivity patterns ofC. jejuni isolates were determined by Kirby Bauer Disc Diffusion assay. These isolates were then subjected to virulence profiling for the detection ofmapA (membrane-associated protein),cadF (fibronectin binding protein),wlaN (beta-1,3-galactosyltransferase) andneuAB (sialic acid biosynthesis gene). FurtherC. jejuni isolates were grouped by random amplification of polymorphic DNA (RAPD) profiling.Results: A total of 436 samples from poultry (n=88), cattle (n=216) and humans (n=132) from different locations were collected. Results revealed percentage ofC. jejuni isolates were 35.2% (31/88), 25.0% (54/216) and 11.3% (15/132) among poultry, cattle and clinical human samples respectively. Antibiotic susceptibility results showed that similar resistance patterns to cephalothin was ie. 87.0%, 87.1% and 89%among humans, poultry and cattle respectively, followed by sulfamethoxazole+trimethoprim 40.0%, 38.7% and 31.0% in humans, poultry and cattle and Ampicillin 40%, 32% and 20% in humans, poultry and cattle respectively. Beta-lactamase activity was detected in 40.00% humans, 20.37% cattle and 32.25% in poultryC. jejuni isolates. CadF andmapA were present in all poultry, cattle and humanC. jejuni isolates,wlaN was not detected in any isolate andneuAB was found in 9/31 (36%) poultry isolates. RAPD profiling results suggested high diversity ofC. jejuni isolates.Conclusions:Detection of multidrug resistantC. jejuni strains from poultry and cattle is alarming as they can be potential hazard to humans. Moreover, predominant association of virulence factors,cadF andmapA (100 % each) inC. jejuni isolates from all sources andneuAB (36%) with poultry isolates suggest the potential source of transmission of diverse types ofC. jejuni to humans.

  4. Genomic characterization of the Guillain-Barre syndrome-associated Campylobacter jejuni ICDCCJ07001 Isolate.

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    Maojun Zhang

    Full Text Available Campylobacter jejuni ICDCCJ07001 (HS:41, ST2993 was isolated from a Guillain-Barré syndrome (GBS patient during a 36-case GBS outbreak triggered by C. jejuni infections in north China in 2007. Sequence analysis revealed that the ICDCCJ07001 genome consisted of 1,664,840 base pairs (bp and one tetracycline resistance plasmid of 44,084 bp. The GC content was 59.29% and 1,579 and 37 CDSs were identified on the chromosome and plasmid, respectively. The ICDCCJ07001 genome was compared to C. jejuni subsp. jejuni strains 81-176, 81116, NCTC11168, RM1221 and C. jejuni subsp. doylei 269.97. The length and organization of ICDCCJ07001 was similar to that of NCTC11168, 81-176 and 81-116 except that CMLP1 had a reverse orientation in strain ICDCCJ07001. Comparative genomic analyses were also carried out between GBS-associated C. jejuni strains. Thirteen common genes were present in four GBS-associated strains and 9 genes mapped to the LOS cluster and the ICDCCJ07001_pTet (44 kb plasmid was mosaic in structure. Thirty-seven predicted CDS in ICDCCJ07001_pTet were homologous to genes present in three virulence-associated plasmids in Campylobacter: 81-176_pTet, pCC31 and 81-176_pVir. Comparative analysis of virulence loci and virulence-associated genes indicated that the LOS biosynthesis loci of ICDCCJ07001 belonged to type A, previously reported to be associated with cases of GBS. The polysaccharide capsular biosynthesis (CPS loci and the flagella modification (FM loci of ICDCCJ07001 were similar to corresponding sequences of strain 260.94 of similar serotype as strain ICDCCJ07001. Other virulence-associated genes including cadF, peb1, jlpA, cdt and ciaB were conserved between the C. jejuni strains examined.

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

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    Valentina Bianchini

    2014-07-01

    Full Text Available 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.

  6. Genotypes and antibiotic resistances of Campylobacter jejuni isolates from cattle and pigeons in dairy farms.

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

    2014-07-01

    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

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

    Science.gov (United States)

    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

  8. PCR detection of seven virulence and toxin genes of Campylobacter jejuni and Campylobacter coli isolates from Danish pigs and cattle and cytolethal distending toxin production of the isolates

    DEFF Research Database (Denmark)

    Bang, Dang Duong; Nielsen, E.M.; Scheutz, F.;

    2003-01-01

    Aims: To study the prevalence of seven virulence and toxin genes, and cytolethal distending toxin (CDT) production of Campylobacter jejuni and C. coli isolates from Danish pigs and cattle. Methods and Results: The presence of the cadF, ceuE, virB11, flaA, cdtA, cdtB, cdtC and the cdt gene cluster...

  9. Comparison of Characteristics of Patients Infected by Campylobacter jejuni, Campylobacter coli, and Campylobacter fetus

    OpenAIRE

    Bessède, Emilie; Lehours, Philippe; Labadi, Leila; BAKIRI, Sarah; Mégraud, Francis

    2014-01-01

    A large database of Campylobacter isolates precisely identified at the species level was used to compare patients' characteristics. In a multivariate analysis, Campylobacter coli was found more often in older patients and in patients having traveled abroad and less often in summertime than Campylobacter jejuni. Campylobacter fetus infection occurred in much older patients and in hospitalized patients with a systemic disease.

  10. Isolation and enumeration of Campylobacter jejuni from poultry products by a selective enrichment method.

    OpenAIRE

    Wesley, R D; Swaminathan, B; Stadelman, W. J.

    1983-01-01

    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 an...

  11. Multilocus Sequence Typing and Antimicrobial Resistance of Campylobacter jejuni Isolated from Dairy Calves in Austria

    OpenAIRE

    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 pr...

  12. Comparative Analysis of Cultural Isolation and Pcr Based Assay for Detection of Campylobacter Jejuni In Food and Faecal Samples

    OpenAIRE

    Singh, Harkanwaldeep; Rathore, R. S.; Singh, Satparkash; Cheema, Pawanjit Singh

    2011-01-01

    In the present study, the efficacy of polymerase chain reaction (PCR) based on mapA gene of C. jejuni was tested for detection of Campylobacter jejuni in naturally infected as well as spiked faecal and food samples of human and animal origin. Simultaneously, all the samples were subjected to the cultural isolation of organism and biochemical characterization. The positive samples resulted in the amplification of a DNA fragment of size ~589 bp in PCR assay whereas the absence of such amplicon ...

  13. Comparative analysis of cultural isolation and PCR based assay for detection of Campylobacter jejuni in food and faecal samples

    OpenAIRE

    Harkanwaldeep Singh; Rathore, R. S.; Satparkash Singh; Pawanjit Singh Cheema

    2011-01-01

    In the present study, the efficacy of polymerase chain reaction (PCR) based on mapA gene of C. jejuni was tested for detection of Campylobacter jejuni in naturally infected as well as spiked faecal and food samples of human and animal origin. Simultaneously, all the samples were subjected to the cultural isolation of organism and biochemical characterization. The positive samples resulted in the amplification of a DNA fragment of size ~589 bp in PCR assay whereas the absence of such amplicon ...

  14. Multilocus sequence typing and antimicrobial resistance of Campylobacter jejuni isolated from dairy calves in Austria

    Directory of Open Access Journals (Sweden)

    Daniela eKlein-Jöbstl

    2016-02-01

    Full Text Available 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 versus conventional and calf housing (place, and individual versus 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 with (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 Campylobacter jejuni and pose a risk for transmission, including antimicrobial resistant isolates to the environment and to humans.

  15. Campylobacter jejuni in commercial eggs

    Directory of Open Access Journals (Sweden)

    Belchiolina Beatriz Fonseca

    2014-01-01

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

  16. Isolation and enumeration of Campylobacter jejuni from poultry products by a selective enrichment method.

    Science.gov (United States)

    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

  17. Occurrence of Campylobacter jejuni and Campylobacter coli and their biotypes in beef and dairy cattle from the south of Chile

    OpenAIRE

    Heriberto Fernández; Marianne Hitschfeld

    2009-01-01

    The prevalence of Campylobacter jejuni and Campylobacter coli and their biotypes in beef and dairy cattle from the South of Chile was established. Campylobacter were statistically more prevalent among beef cattle (35.9%) than among dairy cattle (21.3%), being C. jejuni the species most frequently isolated.Foi estabelecida a prevalência de Campylobacter jejuni e Campylobacter coli e seus biotipos, em bovinos de corte e de leite do sul do Chile. Campylobacter foi estatisticamente mais prevalent...

  18. Effect of human isolated probiotic bacteria on preventing Campylobacter jejuni colonization of poultry.

    Science.gov (United States)

    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.

  19. Septic abortion caused by Campylobacter jejuni bacteraemia.

    Science.gov (United States)

    Skuhala, Tomislava; Škerk, Višnja; Markotić, Alemka; Bukovski, Suzana; Desnica, Boško

    2016-08-01

    A 20-year-old female patient, 14 weeks pregnant, was admitted to hospital with anamnestic and clinical features of acute pyelonephritis. Clinical signs of septic abortion developed and after obstetric examination the therapy was changed to ampicillin, gentamicin and clindamycin. Campylobacter jejuni was isolated from blood cultures. Pathohistological findings confirmed diagnosis of purulent chorioamnionitis. After 2 weeks of ciprofloxacin administration the patient fully recovered. Campylobacter jejuni was not isolated from stool culture and no signs of acute enteritis were registered during the illness. Invasive forms of Campylobacter disease without enteritis are not unusual in immunocompromised hosts but they are restricted to C. fetus rather than C. jejuni isolates. PMID:25872616

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

    Science.gov (United States)

    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.

  1. Inflammasome activation by Campylobacter jejuni

    NARCIS (Netherlands)

    Bouwman, Lieneke I; de Zoete, Marcel R; Bleumink-Pluym, Nancy M C; Flavell, Richard A; van Putten, Jos P M

    2014-01-01

    The Gram-negative pathogen Campylobacter jejuni is the most common cause of bacterial foodborne disease worldwide. The mechanisms that lead to bacterial invasion of eukaryotic cells and massive intestinal inflammation are still unknown. In this study, we report that C. jejuni infection of mouse macr

  2. Compuesto bactericida contra Campylobacter jejuni

    OpenAIRE

    Gañan, M.; Carrascosa, Alfonso V.; Martínez-Rodríguez, Adolfo J.

    2008-01-01

    Compuesto bactericida contra Campylobacter jejuni. Uso de algunos compuestos fenólicos como agentes antibacterianos contra C jejuni. Además de sus usos para conservación de alimentos, suplemento alimenticio para animales y para la elaboración de una composición farmacéutica para el tratamiento de enfermedades causadas por C. jejuni, debido a su actividad frente a este microorganismos.

  3. Multilocus Sequence Typing and Antimicrobial Resistance of Campylobacter jejuni Isolated from Dairy Calves in Austria.

    Science.gov (United States)

    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 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

  4. Detection of seven virulence and toxin genes of Campylobacter jejuni isolates from Danish turkeys by PCR and cytolethal distending toxin production of the isolates

    DEFF Research Database (Denmark)

    Bang, Dang Duong; Borck, Birgitte; Nielsen, Eva Møller;

    2004-01-01

    A total of 117 Campylobacter jejuni isolates from Danish turkeys were tested for the presence of seven virulence and toxin genes by PCR. One hundred seventeen (100%) isolates were positive for flaA, cadF, and ceuE gene primers. One hundred three (88%) isolates were positive for cdt gene cluster P...

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

    OpenAIRE

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

    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.

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

    Science.gov (United States)

    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

  7. Media for Campylobacter jejuni and other campylobacters

    NARCIS (Netherlands)

    Mossel, D.A.A.

    1985-01-01

    Despite their recent elaboration and the many variations in antibiotic combinations designed to attain selectivity, highly selective liquid and solid culture media for Campylobacter jejuni have proved satisfactory provided they are incubated at about 42°C and in a microaerophilic atmosphere such as

  8. Cepas de Campylobacter jejuni resistentes a quinolonas aisladas de humanos, gallinas y pollos Quinolone resistant Campylobacter jejuni strains isolated from humans and from poultry

    Directory of Open Access Journals (Sweden)

    Rodolfo Notario

    2011-08-01

    Full Text Available Se compararon 8 aislamientos de Campylobacter jejuni provenientes de humanos con enfermedad diarreica aguda, con 23 aislamientos de cloaca de gallinas y pollos obtenidos de zonas próximas a la ciudad de Rosario, todos resistentes a la ciprofloxacina. Las muestras se sembraron en agar selectivo y se incubaron en microaerofilia a 42 °C. Las colonias se identificaron con el método tradicional. Los aislamientos se conservaron a -70 °C en caldo cerebro corazón con 17% v/v de glicerina. La clonalidad se determinó por RAPD-PCR, utilizando el primer 1254 (Stern NJ. Se interpretaron los aislamientos como clones distintos cuando diferían en una banda de amplificación. Se obtuvieron 5 clones diferentes. Los patrones I, II y V fueron aislados en criaderos industriales de pollos y en humanos (el II también en un establecimiento de gallinas ponedoras de huevos. En un gallinero familiar se obtuvo el patrón I. El patrón III sólo se obtuvo de humanos. El patrón IV se halló en uno de los criaderos pero no en humanos. Se pudo determinar que 93.5% de las cepas se aislaron tanto de animales como de humanos, por lo que se considera posible que la colonización de criaderos con cepas resistentes a los antimicrobianos pudiera ser el origen de la infección de humanos.Eight quinolone resistant Campylobacter jejuni strains isolated from humans with diarrheal disease were compared with 23 isolates from chicken and from laying hens. Samples were cultured on selective agar in microaerophilia, identified by conventional tests, and conserved in 17% glycerol at -70 °C. Clones were determined by RAPD-PCR employing the 1254 primer (Stern NJ. Five patterns were obtained. Patterns I, II, and V were found in both poultry and human isolates. Pattern I was obtained from poultry in a domestic henhouse. Pattern III was only obtained from humans whereas pattern IV was only obtained from poultry. A 95.3% of clones were found in both, humans and poultry. According to these

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

    Directory of Open Access Journals (Sweden)

    Serichantalergs Oralak

    2010-11-01

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

  10. New, extended biotyping scheme for Campylobacter jejuni, Campylobacter coli, and "Campylobacter laridis".

    OpenAIRE

    Lior, H

    1984-01-01

    A biotyping scheme using improved media and methods for the detection of hippurate hydrolysis, rapid H2S production, and DNA hydrolysis was applied to 1,826 cultures of Campylobacter jejuni, Campylobacter coli and "Campylobacter laridis" isolates from human and nonhuman sources. Four biotypes were identified among C. jejuni: 57.3% of the isolates belonged to biotype I; 36.0%, to biotype II; 4.0%, to biotype III; and 2.7%, to biotype IV. C. coli organisms were differentiated into biotype I (67...

  11. A SpoT polymorphism correlates with chill stress survival and is prevalent in clinical isolates of Campylobacter jejuni

    NARCIS (Netherlands)

    Nierop Groot, M.N.; Boer, de A.G.; Pelt, van W.; Hulst-van Arkel, van der M.C.; Leeuw, P.; Widjaja, H.C.A.; Smits, M.A.; Wal, van der F.J.

    2014-01-01

    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 th

  12. [Antimicrobial Susceptibility and Resistance Mutations in Campylobacter jejuni and C. coli Isolates from Human and Meat Sources].

    Science.gov (United States)

    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

  13. Detecção dos genes da toxina citoletal distensiva em estirpes de Campylobacter jejuni isoladas de carcaças de frangos Detection of cytolethal distending toxin genes in strains of Campylobacter jejuni isolated from broiler carcasses

    Directory of Open Access Journals (Sweden)

    A.F. Carvalho

    2010-10-01

    Full Text Available Foram analisadas 80 amostras de sobrecoxas de frangos de corte resfriados provenientes de feiras livres e hipermercados do município de São Paulo, SP. Treze estirpes de Campylobacter spp. foram isoladas em 10 (12,5% sobrecoxas, sendo cinco amostras originárias de feiras livres e cinco de hipermercados. Onze estirpes foram identificadas como Campylobacter jejuni e duas como Campylobacter coli. As 11 estirpes foram confirmadas como C. jejuni pela PCR do gene da hipuricase (hip, e destas, quatro (36,4% apresentaram os três genes (cdtA, cdtB e cdtC codificantes da toxina citoletal distensiva pela multiplex-PCR, sendo três estirpes provenientes de hipermercados e uma de feira livre. Observou-se a presença de estirpes virulentas de C. jejuni, portadoras do complexo de genes cdt, nas amostras de frango resfriado, não só na linha de abate, mas até o ponto final da cadeia de distribuição, nos dois principais centros de venda a varejo.Eighty samples of refrigerated broiler thighs purchased in street markets and supermarkets in the city of São Paulo, SP, were analyzed. Thirteen Campylobacter spp. strains were isolated in 10 (12.5% thighs, five of them from street market samples and other five from supermarkets. Eleven strains were identified as Campylobacter jejuni and two of them as Campylobacter coli. The 11 strains were confirmed to be C. jejuni using PCR for hippuricase (hip gene. From these, multiplex-PCR showed that four (36.4% strains presented the three genes (cdtA, cdtB, and cdtC encoding cytolethal distending toxin: three strains from supermarket and one from street market samples. These results are important, because they demonstrate the presence of virulent C. jejuni strains in refrigerated broiler thigh samples, not only in the slaughterhouse but in the final point of the distribution chain, at the two most important food retail commercer.

  14. Serotypes and typability of Campylobacter jejuni and Campylobacter coli isolated from poultry products

    DEFF Research Database (Denmark)

    Nielsen, Eva Møller; Nielsen, Niels Ladefoged

    1999-01-01

    nontypable. This rate of nontypable isolates is significantly higher than experienced for isolates from other sources than food products, i.e faecal samples from animals and humans. Subculturing and re-typing of the nontypable isolates improved the typability. After two, five and 10 subcultures 16, six...... and one isolate became typable, respectively. Only three isolates (2%) remained nontypable after 10 subcultures....

  15. Cepas de Campylobacter jejuni resistentes a quinolonas aisladas de humanos, gallinas y pollos Quinolone resistant Campylobacter jejuni strains isolated from humans and from poultry

    OpenAIRE

    Rodolfo Notario; Noemí Borda; Telma Gambandé; Joaquín Bermejo; Adriana Ponessa; Virginia Toledo

    2011-01-01

    Se compararon 8 aislamientos de Campylobacter jejuni provenientes de humanos con enfermedad diarreica aguda, con 23 aislamientos de cloaca de gallinas y pollos obtenidos de zonas próximas a la ciudad de Rosario, todos resistentes a la ciprofloxacina. Las muestras se sembraron en agar selectivo y se incubaron en microaerofilia a 42 °C. Las colonias se identificaron con el método tradicional. Los aislamientos se conservaron a -70 °C en caldo cerebro corazón con 17% v/v de glicerina. La clonalid...

  16. Improved Biotyping Schemes for Campylobacter jejuni and Campylobacter coli

    OpenAIRE

    1985-01-01

    Campylobacter jejuni (20 strains) and Campylobacter coli (12 strains) were assigned to four biovars for each species based on phenotypic tests that were easy to perform and interpret. The resulting biotyping schemes offer a greater degree of distinction among C. jejuni and C. coli strains than any of the other biotyping schemes previously described for these organisms.

  17. Comparative analysis of cultural isolation and PCR based assay for detection of Campylobacter jejuni in food and faecal samples

    Directory of Open Access Journals (Sweden)

    Harkanwaldeep Singh

    2011-03-01

    Full Text Available In the present study, the efficacy of polymerase chain reaction (PCR based on mapA gene of C. jejuni was tested for detection of Campylobacter jejuni in naturally infected as well as spiked faecal and food samples of human and animal origin. Simultaneously, all the samples were subjected to the cultural isolation of organism and biochemical characterization. The positive samples resulted in the amplification of a DNA fragment of size ~589 bp in PCR assay whereas the absence of such amplicon in DNA extracted from E. coli, Listeria, Salmonella and Staphylococcus confirmed the specificity of the primers. Of randomly collected 143 faecal samples comprising human diarrheic stools (43, cattle diarrheic faeces (48 and poultry faecal swabs (52 only 4, 3 and 8, respectively, could be detected by isolation whereas 6, 3 and 10, respectively, were found positive by PCR. However, among food samples viz. beef (30, milk (35, cheese (30, only one beef sample was detected both by culture as well as PCR. Additionally, PCR was found to be more sensitive for C. jejuni detection in spiked faecal and food samples (96.1% each as relative to culture isolation which could detect the organism in 86.7% and 80% samples, respectively. The results depicted the superior efficacy of PCR for rapid screening of samples owing to its high sensitivity, specificity and automation potential.

  18. 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

    Science.gov (United States)

    CABALLERO, Moisés; RIVERA, Isabel; JARA, Luis M.; ULLOA-STANOJLOVIC, Francisco M.; SHIVA, Carlos

    2015-01-01

    SUMMARY Feral pigeons (Columbia livia) live in close contact with humans and other animals. They can transmit potentially pathogenic and zoonotic agents. The objective of this study was to isolate and detect strains of diarrheagenic Escherichia coli and Campylobacter jejuni of urban feral pigeons from an area of Lima, Peru. Fresh dropping samples from urban parks were collected for microbiological isolation of E. coli strains in selective agar, and Campylobacter by filtration method. Molecular identification of diarrheagenic pathotypes of E.coli and Campylobacter jejuni was performed by PCR. Twenty-two parks were sampled and 16 colonies of Campylobacter spp. were isolated. The 100% of isolates were identified as Campylobacter jejuni. Furthermore, 102 colonies of E. coliwere isolated and the 5.88% resulted as Enteropathogenic (EPEC) type and 0.98% as Shiga toxin-producing E. coli (STEC). The urban feral pigeons of Lima in Peru can act as a reservoir or carriers of zoonotic potentially pathogenic enteric agents. PMID:26603225

  19. Quinolone and macrolide resistance in Campylobacter jejuni and C-coli: Resistance mechanisms and trends in human isolates

    DEFF Research Database (Denmark)

    Engberg, J.; Aarestrup, Frank Møller; Taylor, D. E.;

    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...... maintained, but fluoroquinolones may now be of limited use in the empiric treatment of Campylobacter infections in many regions....

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

    DEFF Research Database (Denmark)

    Ramonaite, Sigita; Kudirkiene, Egle; Tamuleviciene, Egle;

    2014-01-01

    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 campylobacterio......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...

  1. Intestinal carriage of Campylobacter jejuni and Campylobacter coli among cattle from South-western Norway and comparative genotyping of bovine and human isolates by amplified-fragment length polymorphism

    Directory of Open Access Journals (Sweden)

    Vardund T

    2006-06-01

    Full Text Available Abstract In a survey conducted in 1999–2001, the carriage of thermotolerant Campylobacters in cattle was investigated, and the genetic diversity of C. jejuni within one herd was examined and compared with human isolates. C. jejuni, C. coli and other thermotolerant Campylobacter spp. were isolated from intestinal contents from 26%, 3% and 2% of 804 cattle, respectively. The carriage rate was higher in calves (46% than in adults (29%. Twenty-nine C. jejuni isolates from one herd and 31 human isolates from the study area were genotyped with amplified-fragment length polymorphism (AFLP. Eighty-three % of the bovine isolates fell into three distinct clusters with 95–100% similarity, persistent in the herd for 5–10 months. Among human isolates, 58% showed >90% similarity with bovine isolates. The results show that cattle are a significant and stable reservoir for C. jejuni in the study area. Transmission between individuals within the herd may be sufficient to maintain a steady C. jejuni population independent of environmental influx. The results of this study have provided new information on C. jejuni and C. coli transmission, and also on the carriage in cattle, genotypes stability and similarity between bovine and human isolates.

  2. Monoclonal Antibodies Specific for Hippurate Hydrolase of Campylobacter jejuni

    OpenAIRE

    Steele, Marina; Gyles, Carlton; Chan, Voon Loong; Odumeru, Joseph

    2002-01-01

    Eleven monoclonal antibodies raised against recombinant Campylobacter jejuni hippurate hydrolase were tested for binding to lysates from 19 C. jejuni strains, 12 other Campylobacter strains, and 21 non-Campylobacter strains. Several monoclonal antibodies bound to C. jejuni but not to other Campylobacter species and may be useful in a species-specific immunoassay.

  3. Oxygen requirement and tolerance of Campylobacter jejuni.

    Science.gov (United States)

    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 ...

  4. Campylobacter jejuni--an emerging foodborne pathogen.

    OpenAIRE

    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....

  5. Multi drug resistance of campylobacter jejuni and campylobacter coli to tested antibiotics in strains originating from humans, poultry and swine

    Directory of Open Access Journals (Sweden)

    Tambur Zoran Ž.

    2010-01-01

    Full Text Available Thermophilic Campylobacter are among the most common cause of bacterial enteritis in humans. Food animals are considered one of the most important sources of Campylobacter causing infections in man. Campylobacter infection is clinically mild and resolves spontaneously. In severe or long-lasting cases, treatment with antibiotics is necessary. Resistance of Campylobacter spp. to drugs used in treatment of infection is a matter of concern. The aim of this paper is to determine presence of multi drug resistant strains of Campylobacter jejuni and Campylobacter coli isolated from animals and man. Material for testing was obtained by scraping the cecum surface from boilers, pig cecum and colon, and human feces. For isolation Campylobacter jejuni and Campylobacter coli microaerophilic conditions, temperature of 42°C and antibiotic supplement were required to inhibit the growth of other intestinal bacteria. In this research, for sensitivity testing of Campylobacter jejuni and Campylobacter coli three different methods were used: disc diffusion test, E-test, and dilution agar method. A total of 55 strains of Campylobacter jejuni and Campylobacter coli. Out of the total, 24 strains originated from man, 16 from broilers were isolated, and 15 from pigs. Multidrug resistance was determined in cases when the strains were resistant to two or more antibiotics. Applying E-test, we detected that the largest number of Campylobacter jejuni were multi drug resistant to two antibiotics (41.2%, and three antibiotics (11.8%. Applying disc diffusion method it was detected that 5.9% of Campylobacter jejuni from man was resistant to four tested antibiotics. Applying all three methods, it was detected that the largest number of Campylobacter strains was resistant to two antibiotics and three antibiotics. Applying disc diffusion method it was detected that 50% of Campylobacter coli strains from pigs were resistant to three tested antibiotics.

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

    Science.gov (United States)

    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, ...

  7. Epidemiological aspects of Campylobacter jejuni enteritis.

    OpenAIRE

    Norkrans, G.; Svedhem, A

    1982-01-01

    An epidemiological study on Campylobacter jejuni enterocolitis was performed in an urban Swedish community. The study included 55 patients gathered during a six-month period. Forty-one of the 55 patients (75%) were infected outside Sweden. Campylobacter enterocolitis was rare among children within the country. Patients infected in Sweden had eaten chicken significantly more often than a corresponding control group. Seven out of nine chicken consuming campylobacter patients also had prepared t...

  8. Prevalence of Campylobacter jejuni in poultry breeder flocks

    Directory of Open Access Journals (Sweden)

    Ludovico Dipineto

    2010-01-01

    Full Text Available The aim of this work is to present the preliminary results of a study about the prevalence of Campylobacter jejuni in poultry breeder flocks. It was examined three different breeder flocks of Bojano in Molise region. A total of 360 cloacal swabs and 80 enviromental swabs was collected. Of the 3 flocks studied, 6.9% tested were positive for Campylobacter spp. The most-prevalent isolated species is C. jejuni (8.2%. Only 3 of the 360 cloacal swabs samples examined were associated with C. coli. The environmental swabs resulted negative. This results confirms again that poultry is a reservoir of this germ.

  9. The role of Campylobacter jejuni cytolethal distending toxin in gastroenteritis

    DEFF Research Database (Denmark)

    Mortensen, Ninell P; Schiellerup, Peter; 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, cdt......ABC, 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...

  10. Antibiotic resistance profile and RAPD analysis of Campylobacter jejuni isolated from vegetables farms and retail markets

    Directory of Open Access Journals (Sweden)

    John Yew Huat Tang

    2016-01-01

    Conclusions: C. jejuni contamination in vegetables at retail markets was due to cross contamination. Current finding proved that C. jejuni in small scale vegetables production was less expose towards antibiotic abuse.

  11. Complete Genome Sequences of Campylobacter jejuni Strains OD267 and WP2202 Isolated from Retail Chicken Livers and Gizzards Reveal the Presence of Novel 116-Kilobase and 119-Kilobase Megaplasmids with Type VI Secretion Systems

    Science.gov (United States)

    Marasini, Daya

    2016-01-01

    Genome sequences of Campylobacter jejuni strains OD267 and WP2202, isolated from chicken livers and gizzards, showed the presence of novel 116-kb and 119-kb megaplasmids, respectively. The two megaplasmids carry a type VI secretion system and tetracycline resistance genes. These are the largest sequenced Campylobacter plasmids to date. PMID:27688318

  12. Complete Genome Sequences of Campylobacter jejuni Strains OD267 and WP2202 Isolated from Retail Chicken Livers and Gizzards Reveal the Presence of Novel 116-Kilobase and 119-Kilobase Megaplasmids with Type VI Secretion Systems.

    Science.gov (United States)

    Marasini, Daya; Fakhr, Mohamed K

    2016-01-01

    Genome sequences of Campylobacter jejuni strains OD267 and WP2202, isolated from chicken livers and gizzards, showed the presence of novel 116-kb and 119-kb megaplasmids, respectively. The two megaplasmids carry a type VI secretion system and tetracycline resistance genes. These are the largest sequenced Campylobacter plasmids to date. PMID:27688318

  13. Tracing isolates from domestic human Campylobacter jejuni infections to chicken slaughter batches and swimming water using whole-genome multilocus sequence typing.

    Science.gov (United States)

    Kovanen, Sara; Kivistö, Rauni; Llarena, Ann-Katrin; Zhang, Ji; Kärkkäinen, Ulla-Maija; Tuuminen, Tamara; Uksila, Jaakko; Hakkinen, Marjaana; Rossi, Mirko; Hänninen, Marja-Liisa

    2016-06-01

    Campylobacter jejuni is the leading cause of bacterial gastroenteritis and chicken is considered a major reservoir and source of human campylobacteriosis. In this study, we investigated temporally related Finnish human (n=95), chicken (n=83) and swimming water (n=20) C. jejuni isolates collected during the seasonal peak in 2012 using multilocus sequence typing (MLST) and whole-genome MLST (wgMLST). Our objective was to trace domestic human C. jejuni infections to C. jejuni isolates from chicken slaughter batches and swimming water. At MLST level, 79% of the sequence types (STs) of the human isolates overlapped with chicken STs suggesting chicken as an important reservoir. Four STs, the ST-45, ST-230, ST-267 and ST-677, covered 75% of the human and 64% of the chicken isolates. In addition, 50% of the swimming water isolates comprised ST-45, ST-230 and ST-677. Further wgMLST analysis of the isolates within STs, accounting their temporal relationship, revealed that 22 of the human isolates (24%) were traceable back to C. jejuni positive chicken slaughter batches. None of the human isolates were traced back to swimming water, which was rather sporadically sampled. The highly discriminatory wgMLST, together with the patient background information and temporal relationship data with possible sources, offers a new, accurate approach to trace back the origin of domestic campylobacteriosis. Our results suggest that potentially a substantial proportion of campylobacteriosis cases during the seasonal peak most probably are due to other sources than chicken meat consumption. These findings warrant further wgMLST-based studies to reassess the role of other reservoirs in the Campylobacter epidemiology both in Finland and elsewhere.

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

    Science.gov (United States)

    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

  15. Metronidazole resistance in Campylobacter jejuni from poultry meat

    DEFF Research Database (Denmark)

    Andersen, Sigrid Rita; Shukri, Naseer Mahmoud; Boel, Jeppe;

    2006-01-01

    The occurrence of metronidazole resistance was investigated among Campylobacter jejuni in raw poultry meat collected from supermarkets. MICs were determined by the agar dilution procedure in the testing range of 3 to 60 mu g/ml metronidazole. The MICs showed a bimodal distribution with a signific......The occurrence of metronidazole resistance was investigated among Campylobacter jejuni in raw poultry meat collected from supermarkets. MICs were determined by the agar dilution procedure in the testing range of 3 to 60 mu g/ml metronidazole. The MICs showed a bimodal distribution...... with a significant proportion of metronidazole-resistant isolates among C. jejuni from raw broiler and turkey meat. Metronidazole resistance occurred most frequently among turkey meat isolates (P bacteria carrying metronidazole resistance....

  16. A Charcoal- and Blood-Free Enrichment Broth for Isolation and PCR Detection of Campylobacter jejuni and Campylobacter coli

    Science.gov (United States)

    Campylobacter spp. is a Gram negative bacterium and is the major cause of foodborne gastroenteritis worldwide. The microaerophilic nature of Campylobacter and its requirement of ~5% O2 for growth have complicated its recovery from foods. This is achieved with the addition to the enrichment media of ...

  17. Nutrient Acquisition and Metabolism by Campylobacter jejuni

    Directory of Open Access Journals (Sweden)

    Martin eStahl

    2012-02-01

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

  18. Comparison of genotypes and antibiotic resistances of Campylobacter jejuni and Campylobacter coli on chicken retail meat and at slaughter

    OpenAIRE

    Kittl, Sonja; Korczak, Bozena M.; Niederer, Lilian; Baumgartner, Andreas; Buettner, Sabina; Overesch, Gudrun; Kuhnert, Peter

    2013-01-01

    Multilocus sequence typing (MLST) and antibiotic resistance patterns of Campylobacter jejuni and Campylobacter coli from retail chicken meat showed high overlap with isolates collected at slaughterhouses, indicating little selection along the production chain. They also showed significant common sequence types with human clinical isolates, revealing chicken meat as a likely source for human infection.

  19. Comparison of genotypes and antibiotic resistances of Campylobacter jejuni and Campylobacter coli on chicken retail meat and at slaughter.

    Science.gov (United States)

    Kittl, Sonja; Korczak, Bożena M; Niederer, Lilian; Baumgartner, Andreas; Buettner, Sabina; Overesch, Gudrun; Kuhnert, Peter

    2013-06-01

    Multilocus sequence typing (MLST) and antibiotic resistance patterns of Campylobacter jejuni and Campylobacter coli from retail chicken meat showed high overlap with isolates collected at slaughterhouses, indicating little selection along the production chain. They also showed significant common sequence types with human clinical isolates, revealing chicken meat as a likely source for human infection. PMID:23584778

  20. Resistance to quinolones in Campylobacter jejuni and Campylobacter coli from Danish broilers at farm level

    DEFF Research Database (Denmark)

    Pedersen, Karl; Wedderkopp, A.

    2003-01-01

    Aims : To investigate the prevalence of quinolone resistance among Campylobacter jejuni and Camp. coli isolates from Danish poultry at the farm level, as well as for the whole country. Methods and Results : Data and isolates were collected from a national surveillance of Campylobacter in poultry....... Quinolone resistance was investigated by determination of minimum inhibitory concentration (MIC) to nalidixic acid and enrofloxacin. Among Camp. jejuni and Camp. coli combined, 7.5% were resistant to nalidixic acid. Quinolone resistance varied considerably from farm to farm, with 0% on some farms and almost...... 100% on others, but the resistance was evenly distributed geographically. With respect to isolates from farms where resistance was detected, quinolone resistance was higher among Camp. coli (28.7%) than among Camp. jejuni (11.3%). PFGE typing of quinolone-resistant and quinolone-susceptible isolates...

  1. A comparative analysis of methylome profiles of Campylobacter jejuni sheep abortion isolate and gastroenteric strains using PacBio data

    Directory of Open Access Journals (Sweden)

    Kathy T Mou

    2015-01-01

    Full Text Available Campylobacter jejuni is a leading cause of human gastrointestinal disease and small ruminant abortions in the United States. The recent emergence of a highly virulent, tetracycline-resistant C. jejuni subsp. jejuni sheep abortion clone (clone SA in the United States, and that strain’s association with human disease, has resulted in a heightened awareness of the zoonotic potential of this organism. Pacific Biosciences’ Single Molecule, Real-Time sequencing technology was used to explore the variation in the genome-wide methylation patterns of the abortifacient clone SA (IA3902 and phenotypically distinct gastrointestinal-specific C. jejuni strains (NCTC 11168 and 81-176. Several notable differences were discovered that distinguished the methylome of IA3902 from that of 11168 and 81-176: identification of motifs novel to IA3902, genome-specific hypo- and hypermethylated regions, strain level variability in genes methylated, and differences in the types of methylation motifs present in each strain. These observations suggest a possible role of methylation in the contrasting disease presentations of these three C. jejuni strains. In addition, the methylation profiles between IA3902 and a luxS mutant were explored to determine if variations in methylation patterns could be identified that might explain the role of LuxS-dependent methyl recycling in IA3902 abortifacient potential.

  2. An outbreak of infectious hepatitis in commercially reared ostriches associated with Campylobacter coli and Campylobacter jejuni

    DEFF Research Database (Denmark)

    Stephens, C.P.; On, S.L.W.; Gibson, J.A.

    1998-01-01

    A disease causing high morbidity and mortality was observed in young ostriches from six properties in southeast Queensland, Australia. The disease affected birds from 2-8 weeks of age and was characterised clinically by bright-green urates and pathologically by severe necrotic hepatitis. The liver...... lesions resembled those of vibrionic hepatitis in other avian species. Campylobacter coli was isolated from the livers of affected ostriches from five of the six properties. Campylobacter jejuni subsp. jejuni was isolated from birds from the remaining property. Pulsed-field gel electrophoresis-based (PFGE...... biochemical test results and PFGE clearly distinguished the C. jejuni strain isolated from the geographically remote farm from the outbreak of C. coli type. We believe this to be the first definitive report of avian hepatitis associated with C. coli....

  3. Genomic Characterization of Campylobacter jejuni strain M1

    DEFF Research Database (Denmark)

    Friis, Carsten; Wassenaar, Gertrude Maria; Javed, Muhammad A.;

    2010-01-01

    Campylobacter jejuni strain M1 (laboratory designation 99/308) is a rarely documented case of direct transmission of C. jejuni from chicken to a person, resulting in enteritis. We have sequenced the genome of C. jejuni strain M1, and compared this to 12 other C. jejuni sequenced genomes currently...

  4. Comparative genomic assessment of Multi-Locus Sequence Typing: rapid accumulation of genomic heterogeneity among clonal isolates of Campylobacter jejuni

    Directory of Open Access Journals (Sweden)

    Nash John HE

    2008-08-01

    Full Text Available Abstract Background Multi-Locus Sequence Typing (MLST has emerged as a leading molecular typing method owing to its high ability to discriminate among bacterial isolates, the relative ease with which data acquisition and analysis can be standardized, and the high portability of the resulting sequence data. While MLST has been successfully applied to the study of the population structure for a number of different bacterial species, it has also provided compelling evidence for high rates of recombination in some species. We have analyzed a set of Campylobacter jejuni strains using MLST and Comparative Genomic Hybridization (CGH on a full-genome microarray in order to determine whether recombination and high levels of genomic mosaicism adversely affect the inference of strain relationships based on the analysis of a restricted number of genetic loci. Results Our results indicate that, in general, there is significant concordance between strain relationships established by MLST and those based on shared gene content as established by CGH. While MLST has significant predictive power with respect to overall genome similarity of isolates, we also found evidence for significant differences in genomic content among strains that would otherwise appear to be highly related based on their MLST profiles. Conclusion The extensive genomic mosaicism between closely related strains has important implications in the context of establishing strain to strain relationships because it suggests that the exact gene content of strains, and by extension their phenotype, is less likely to be "predicted" based on a small number of typing loci. This in turn suggests that a greater emphasis should be placed on analyzing genes of clinical interest as we forge ahead with the next generation of molecular typing methods.

  5. Characterization of the virulence, growth temperature and antibiotic resistance of the Campylobacter jejuni IAL 2383 strain isolated from humans

    Directory of Open Access Journals (Sweden)

    B.B. Fonseca

    2014-01-01

    Full Text Available The objective of this study was to characterize the C. jejuni IAL2383 strain isolated from humans in Brazil. Transcripts for the racR, dnaJ and ciaB genes were found and flaA, plda and cadF genes were present in the genome and bacteria was sensitive to most of the important antimicrobials used to treat humans. C. jejuni IAL2383 is a good experimental model to analyze the interactions with cells.

  6. Genetic relatedness among Campylobacter jejuni serotyped isolates of diverse origin as determined by numerical analysis of amplified fragment length polymorphism (AFLP) profiles

    DEFF Research Database (Denmark)

    Siemer, B.L.; Harrington, C.S.; Nielsen, E.M.;

    2004-01-01

    Aims: To use amplified fragment length polymorphism (AFLP) analysis to evaluate the genetic relatedness among 254 Campylobacter jejuni reference and field strains of diverse origin representing all defined 'Penner' serotypes for this species. Methods and Results: Field strains (n = 207) from human...... health. The remaining 30 groups contained isolates from humans, chickens and associated food products, cattle, sheep, turkeys, ostriches and/or dogs. Strains assigned to serotypes 2, 6/7, 11 and 12 formed major clusters at the 77.6% S-level. Most other serotypes did not form homogeneous clusters...

  7. Isolation of Campylobacter jejuni from cloaca and cecum content of chicken broilers bred in intensive systems in the Western part of Romani

    Directory of Open Access Journals (Sweden)

    Ada Cean

    2013-10-01

    Full Text Available Campylobacter spp., belongs to the group of thermo-tolerant bacteria, and is the most frequent cause of gastrointestinal diseases in humans following consumption of poorly cooked chicken meat. The aim of our study was to test the common methodology for isolation of Campylobacter jejuni species from cloaca and cecum content of chicken broilers breed in intensive systems in Western part of Romania. The experiments were conducted during July –September 2013. As biological material we used chicken broilers from 6 intensive breeding facilities from the West part of Romania, from which cloaca swabs and cecum content were recovered as samples. Bacteria isolation was performed by inseminating Petri dish with Muller Hinton Agar media, after bacterial growth, they were subculture on Muller-Hinton Agar with Skirrow. The bacteria were tested by Gram staining and Oxidase test. Bacterial growth was detected from all samples when grown on Mueller-Hinton Agar, but when the bacteria was passed on Muller Hinton Agar with selective supplement (Skirrow 27 out of 36 samples remained positive (75,0%. With respect to the sample origin 13 (72.2% samples from cloaca swab and 14 (77.7% from cecum content grown on campylobacter selective media. All samples from Muller-Hinton supplemented with Skirrow tested negative for Gram staining and positive for oxidase test. We have successfully isolated Campylobacter spp., strains from farms and private producers in the western part of Romania.

  8. Comparison of electrophoretic protein profiles of Campylobacter jejuni subsp. jejuni isolated from different animal species Comparação dos perfis eletroforéticos de proteínas de Campylobacter jejuni subsp. jejuni isoladas de diferentes espécies animais

    Directory of Open Access Journals (Sweden)

    Eliana Scarcelli

    2001-12-01

    Full Text Available Electrophoretic protein profiles of Campylobacter jejuni subsp. jejuni strains isolated from feces of seven animal species, including man, were compared. Fourteen strains (two from each species plus two human strains and the reference one, were ruptured by ultrasound and their total soluble proteins were analyzed by SDS-PAGE technique in a 12% polyacrylamide gel with computerized densitometric reading by the molecular analyst software. All the strains had bands in common that correspond to 45 and 66 Kda molecular weight. The disagreement corresponded to a 97 to 200 Kda molecular weight region. From the 17 strains, 13 (76.5%, were classified as biotype I, three (17.6% as biotype II and one (5.8% as biotype III. Since protein extracts were obtained from cells grown under identical conditions, and thus, able to express the same phenotype, this disagreement region could be related to different genotypes or serotypes.Perfis eletroforéticos de proteínas de cepas Campylobacter jejuni subsp. jejuni isoladas de fezes de diferentes espécies animais, inclusive o homem, foram comparados. Quatorze cepas (duas de cada espécie mais duas cepas de origem humana e a cepa de referência foram rompidas por ultra-som e suas proteínas solúveis totais analisadas através das técnicas de SDS-PAGE em gel de poliacrilamida a 12% e análise densitométrica. Todas as cepas tinham em comum bandas que migraram em regiões que correspondiam ao peso molecular de 45 e 66 Kda. As regiões discordantes correspondiam principalmente às regiões entre 97 e 200 Kda. Das 17 cepas, 13 (76.5%, foram classificadas como biotipo I, três (17.6% como biotipo II e uma (5.8% como biotipo III. Uma vez que os extratos de proteínas foram obtidos de células que se desenvolveram sob condições idênticas, possibilitando a expressão do mesmo fenótipo, estas regiões protéicas discordantes poderiam estar relacionadas a diferentes sorotipos ou genótipos.

  9. Epidemiological aspects of Campylobacter jejuni enteritis.

    Science.gov (United States)

    Norkrans, G; Svedhem, A

    1982-08-01

    An epidemiological study on Campylobacter jejuni enterocolitis was performed in an urban Swedish community. The study included 55 patients gathered during a six-month period. Forty-one of the 55 patients (75%) were infected outside Sweden. Campylobacter enterocolitis was rare among children within the country. Patients infected in Sweden had eaten chicken significantly more often than a corresponding control group. Seven out of nine chicken consuming campylobacter patients also had prepared the fresh chicken alone, and none of their family members became ill. Thus the preparation of food contaminated with Campylobacter seems to elevate the risk for contracting the disease. Sick household pets transmitted the campylobacter infection to two patients. Forty-six of the patients had a total of 85 close household members. Three definite secondary cases were found. There was no evidence of transmission of Campylobacter by food prepared by two cooks who were working while still being asymptomatic excreters. Clinical reinfection with Campylobacter was observed in one patient. No patients became long-term carriers of Campylobacter. PMID:7097000

  10. Using Major Outer Membrane Protein Typing as an Epidemiological Tool To Investigate Outbreaks Caused by Milk-Borne Campylobacter jejuni Isolates in California

    Science.gov (United States)

    Mandrell, Robert E.; Yuan, Jean; Bates, Anna; Manalac, Rosa; Mohle-Boetani, Janet; Kimura, Akiko; Lidgard, Janice; Miller, William G.

    2013-01-01

    We describe using major outer membrane protein (MOMP) typing as a screen to compare the Campylobacter jejuni porA gene sequences of clinical outbreak strains from human stool with the porA sequences of dairy farm strains isolated during two milk-borne campylobacteriosis outbreak investigations in California. The genetic relatedness of clinical and environmental strains with identical or closely related porA sequences was confirmed by multilocus sequence typing and pulsed-field gel electrophoresis analysis. The first outbreak involved 1,644 C. jejuni infections at 11 state correctional facilities and was associated with consumption of pasteurized milk supplied by an on-site dairy (dairy A) at a prison in the central valley. The second outbreak involved eight confirmed and three suspect C. jejuni cases linked to consumption of commercial raw milk and raw chocolate colostrum at another central valley dairy (dairy B). Both dairies bottled fluid milk on the farm and distributed the finished product to off-site locations. Altogether, C. jejuni was isolated from 7 of 15 (46.7%) bovine fecal, 12 of 20 (60%) flush alley water, and 1 of 20 (5%) lagoon samples collected on dairy A. At dairy B, C. jejuni was cultured from 9 of 26 (34.6%) bovine fecal samples. Environmental strains indistinguishable from the clinical outbreak strains were found in five flush alley water samples (dairy A) and four bovine fecal samples (dairy B). The findings demonstrate that MOMP typing is a useful tool to triage environmental isolates prior to conducting more labor-intensive molecular typing methods. PMID:23115263

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

    Science.gov (United States)

    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...

  12. Trends in occcurrence of antimicrobial resistance in Campylobacter jejuni isolates from broiler chickens, broiler chicken meat, and human domestically acquired cases and travel associated cases ind Denmark

    DEFF Research Database (Denmark)

    Skjøt-Rasmussen, Line; Ethelberg, Steen; Emborg, Hanne-Dorthe;

    2009-01-01

    Campylobacter jejuni is a frequent cause of bacterial gastroenteritis. Often it causes self-limiting disease but severe or prolonged cases may require antimicrobial treatment. The agricultural use of antimicrobial agents selects for resistance among C. jejuni which is transmitted to humans via food...

  13. Campylobacter jejuni y Campylobacter coli en tres grupos de gallinas de diferente origen geográfico del sur de Chile Occurrence of Campylobacter jejuni and Campylobacter coli in three groups of hens of different geographic origin in Southern Chile

    Directory of Open Access Journals (Sweden)

    H. FERNANDEZ

    2000-01-01

    Full Text Available Se estudiaron 300 muestras fecales de gallinas obtenidas en tres sitios geográficos del sur de Chile (comunas de Loncoche, Valdivia y Puerto Montt, para conocer la prevalencia de Campylobacter jejuni y de C. coli en estas aves consideradas como reservorio. La prevalencia de especies termotolerantes del género Campylobacter fue del 25,7%, siendo C. jejuni aislado con una frecuencia del 76,6% y la de C. coli del 23,4%. Fueron encontrados sólo 2 de los 4 biotipos de C. jejuni, siendo aislado el biotipo II con mayor frecuencia (68.8%. En C. coli fueron encontrados los 2 biotipos descritos para esta especie, siendo, también, el biotipo II el más frecuenteIn order to establish the prevalence of Campylobacter jejuni and C. coli, 300 fecal samples obtained from hens in three different geographical places (Loncoche, Valdivia and Puerto Montt cities from Southern Chile were studied. The prevalence of the thermotolerant species of Campylobacter was 25.7%, being Campylobacter jejuni isolated with a frequency of 76.6% and C. coli with 23.4%. Only two of the four biotypes of C. jejuni were found, being biotype II the most frequent one (68.8%. The two biotypes described for C. coli were isolated, and biotype II was also the most frequent (68.8%.

  14. Invasion Assays and Genomotyping to Investigate Differences in Virulence of Campylobacter spp. Isolates from Iceland

    Science.gov (United States)

    Campylobacter spp. are the leading cause of human gastroenteritis worldwide. Epithelial cell invasion is thought to be essential for Campylobacter spp. infection. Previous invasion studies with intestinal epithelial cells revealed that the ability of different Campylobacter jejuni isolates to inva...

  15. Characterisation of Campylobacter jejuni genes potentially involved in phosphonate degradation

    Directory of Open Access Journals (Sweden)

    Hartley Lauren E

    2009-06-01

    Full Text Available Abstract Potential biological roles of the Campylobacter jejuni genes cj0641, cj0774c and cj1663 were investigated. The proteins encoded by these genes showed sequence similarities to the phosphonate utilisation PhnH, K and L gene products of Escherichia coli. The genes cj0641, cj0774c and cj1663 were amplified from the pathogenic C. jejuni strain 81116, sequenced, and cloned into pGEM-T Easy vectors. Recombinant plasmids were used to disrupt each one of the genes by inserting a kanamycin resistance (KmR cassette employing site-directed mutagenesis or inverse PCR. Campylobacter jejuni 81116 isogenic mutants were generated by integration of the mutated genes into the genome of the wild-type strain. The C. jejuni mutants grew on primary isolation plates, but they could not be purified by subsequent passages owing to cell death. The mutant C. jejuni strains survived and proliferated in co-cultures with wild-type bacteria or in media in which wild-type C. jejuni had been previously grown. PCR analyses of mixed wild-type/mutant cultures served to verify the presence of the mutated gene in the genome of a fraction of the total bacterial population. The data suggested that each mutation inactivated a gene essential for survival. Rates of phosphonate catabolism in lysates of E. coli strain DH5α were determined using proton nuclear magnetic resonance spectroscopy. Whole-cell lysates of the wild-type degraded phosphonoacetate, phenylphosphonate and aminomethylphosphonate. Significant differences in the rates of phosphonate degradation were observed between lysates of wild-type E. coli, and of bacteria transformed with each one of the vectors carrying one of the C. jejuni genes, suggesting that these genes were involved in phosphonate catabolism.

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

    Science.gov (United States)

    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 ...

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

    Science.gov (United States)

    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. PMID:26371330

  18. Campylobacter jejuni: exposure assessment and hazard characterization : growth, survival and infectivity of Campylobacter jejuni

    NARCIS (Netherlands)

    Verhoeff-Bakkenes, L.

    2012-01-01

    Campylobacter jejuni, a small, curved or spirally shaped highly motile microorganism, is identified as a major cause of bacterial gastroenteritis throughout the world. Serious complications such as the Guillain-Barré syndrome and reactive arthritis might occasionally follow infection. In this

  19. PCR-based restriction fragment length polymorphism (RFLP) analysis of Campylobacter jejuni isolates from humans, chickens and dogs in northern Taiwan.

    Science.gov (United States)

    Tsai, Hsiang-Jung; Huang, Huang-Chi; Tsai, Huei-Lin; Chang, Cha-Chin

    2006-08-01

    Two hundred and twenty strains of Campylobacter jejuni (70 human, 51 canine and 99 chicken strains) were isolated from September 2003 to September 2004 in northern Taiwan. These strains were subtyped by PCR-RFLP analysis of the flagellin (FlaA) gene. On the basis of restrictive digest, six types were identified with AfaI, seven types with MboI and five types with HaeIII. With the combination of these three enzymes, 47 distinct PCR-RFLP patterns were observed-25 each from human and chicken isolates, and 9 from canine isolates. In human strains, the most frequently occurring types were Cj-28 (14.3%), Cj-17 (10%), Cj-16 (8.6%), Cj-37 (7.1%) and Cj-46 (7.1%). In canine strains, the most prevalent types were Cj-1 (33.3%), Cj-26 (19.6%), Cj-3 (15.7%), Cj-2 (9.8%) and Cj-10 (9.8%). In chicken strains, the most frequently occurring types were Cj-46 (40.4%), Cj-29 (9.1%), Cj-45 (7.1%) and Cj-41 (5.1%). The results suggest that poultry is a source, but not the sole source, of C. jejuni infection in humans. Two RFLP types, Cj-17 and Cj-37, frequently occurring in human isolates in this study have also been found to be prevalent in human isolates in Japan, China and the Czech Republic, indicating a possible international clonal spread.

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

    Science.gov (United States)

    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).

  1. Survival and resuscitation of ten strains of Campylobacter jejuni and Campylobacter coli under acid conditions

    NARCIS (Netherlands)

    Chaveerach, P.; Huurne, ter A.A.H.M.; Lipman, L.J.A.; Knapen, van F.

    2003-01-01

    The culturability of 10 strains of Campylobacter jejuni and Campylobacter coli was studied after the bacteria were exposed to acid conditions for various periods of time. Campylobacter cells could not survive 2 h under acid conditions (formic acid at pH 4). The 10 Campylobacter strains could not be

  2. Effect of gamma radiation on Campylobacter jejuni

    Energy Technology Data Exchange (ETDEWEB)

    Lambert, J.D.; Maxcy, R.B.

    Radiation resistance of Campylobacter jejuni in broth, ground beef, and ground turkey meat was determined using dose levels from 0-200 Krad at -30 +/- 10/sup 0/C, at 0-5/sup 0/C, and at 30 +/- 10/sup 0/C. Irradiation at -30/sup 0/C increased radiation resistance of cultures in ground meats; broth cultures were not greatly influenced by temperature. The effect of culture age on radiation resistance was also evaluated using cells in various physiological phases. Age did not have a pronounced effect on radiation resistance. The largest D/sub 10/ value for C. jejuni was 32 Krad, which was less than D/sub 10/ values commonly reported for salmonellae. 20 references, 4 figures.

  3. Substrate utilization by Campylobacter jejuni and Campylobacter coli

    Energy Technology Data Exchange (ETDEWEB)

    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.

  4. Genome Sequence of Campylobacter jejuni strain 327, a strain isolated from a turkey slaughterhouse

    DEFF Research Database (Denmark)

    Takamiya, Monica; Özen, Asli Ismihan; Rasmussen, Morten;

    2011-01-01

    , catalase positive bacterium obtains energy from the metabolism of amino acids and Krebs cycle intermediates. Strain 327 was isolated from a turkey slaughter production line and is considered environmentally sensitive to food processing (cold, heat, drying) and storage conditions. The 327 whole genome...

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

    Science.gov (United States)

    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.

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

    Directory of Open Access Journals (Sweden)

    Muhammad A Javed

    Full Text Available 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.

  7. Use of culture, PCR analysis, and DNA microarrays for detection of Campylobacter jejuni and Campylobacter coli from chicken feces

    DEFF Research Database (Denmark)

    Keramas, Georgios; Bang, Dang Duong; Lund, Marianne;

    2004-01-01

    . detection obtained with DNA microarrays were compared to those obtained by conventional culture and gel electrophoresis. By conventional culture, 60% of the samples were positive for either Campylobacter jejuni or Campylobacter coli. By PCR and capillary electrophoresis, 95% of the samples were positive....... coli by conventional culture but were positive for C. jejuni by both PCR-capillary electrophoresis and DNA microarray analysis. The discrepancy between the methods is discussed....... for Campylobacter spp., whereas with DNA microarrays all samples were positive for Campylobacter spp. By application of DNA microarray analysis, the isolates in 4 samples (6%) could not be identified to the species level, whereas by PCR-capillary electrophoresis, the isolates in 12 samples (19%) remained...

  8. Evaluation of 11 PCR assays for species-level identification of Campylobacter jejuni and Campylobacter coli

    DEFF Research Database (Denmark)

    On, Stephen L.W.; Jordan, Penelope J.

    2003-01-01

    We examined the sensitivity and specificity of 11 PCR assays described for the species identification of Campylobacter jejuni and Campylobacter coli by using 111 type, reference, and field strains of C. jejuni, C. coli, and Campylobacter lari. For six assays, an additional 21 type strains...... representing related Campylobacter, Arcobacter, and Helicobacter species were also included. PCR tests were initially established in the laboratory by optimizing conditions with respect to five type and reference strains of C. jejuni, C. coli, and C. lari. One PCR test for C. coli failed to give appropriate...

  9. Microbiological diagnosis of infections caused by Campylobacter jejuni and Campylobacter coli in humans

    Directory of Open Access Journals (Sweden)

    Natalia Rokosz

    2014-01-01

    Full Text Available Campylobacter jejuni and Campylobacter coli are Gram-negative, microaerophilic bacteria which are worldwide in distribution, causing a zoonotic disease in humans called campylobacteriosis. These infections are mainly caused by eating contaminated food products, most often improperly prepared poultry meat. Campylobacteriosis usually takes the form of gastroenteritis, or inflammation of the intestines, and the characteristic symptoms are watery-mucous diarrhea often with the presence of blood in stool, nausea, vomiting, abdominal pain and fever. The epidemiological data suggest that in Europe, as well as in North America, bacteria of the genus Campylobacter, especially C. jejuni and C. coli, are the most commonly isolated pathogens in infections of the gastrointestinal tract in humans. Epidemiological data indicate that these organisms are a much more common cause of acute diarrhea, mostly in young children, than Salmonella and Yersinia. The lack of specific symptoms makes the diagnosis of campylobacteriosis necessary to carry out specialized microbiological diagnostics. Because so far these studies are performed in our country only in a few laboratories, the overwhelming number of cases of campylobacteriosis are not recorded in Polish epidemiological statistics. The purpose of this paper is to discuss issues related to the microbiological diagnosis of infections caused by C. jejuni and C. coli. It also describes the basic epidemiological and clinical data, as well as current treatment of campylobacteriosis.

  10. DNA Micorarrays for Genotyping and Population Studies of Campylobacter jejuni

    Science.gov (United States)

    Introduction: Campylobacter jejuni is a major cause worldwide of foodborne bacterial gastroenteritis. The continued development of more effective and informative typing methods is necessary to improve our understanding of the epidemiology and population dynamics of this important pathogen. Comparat...

  11. Prevalence of Campylobacter Jejuni and Coli in Sheep Carcasses by Using

    Directory of Open Access Journals (Sweden)

    Reza Shahrokhabadi

    2013-11-01

    Full Text Available Background: Campylobacter species are common bacterial pathogens causing gastroenteritis in humans worldwide. Materials and Methods: A total of 148 randomly sheep carcasses were sampled by surface section of neck meat taken immediately after slaughter analyzed using microbiological examinations. Results: Campylobacter spp. was isolated from 10.13% meat cultures samples examined. Among these 80% sample were C. jejuni and 20% sample were C. coli. Using PCR assays, the number of positive campylobacters increased to 11.48%. Of these positive samples, 82.35% were C. jejuni and 17.65% were C. coli. Significantly higher prevalence rates of Campylobacter spp. (p<0.05 were found in the meat samples taken in summer (47.05%. Conclusion: The PCR is a reliable and sensitive method which can be used as a diagnostic technique for the detection of campylobacter in lamb samples.

  12. Identification of genomic differences between Campylobacter jejuni subsp. jejuni and C. jejuni subsp. doylei at the nap locus leads to the development of a C. jejuni subspeciation multiplex PCR method

    Directory of Open Access Journals (Sweden)

    Heath Sekou

    2007-02-01

    Full Text Available Abstract Background The human bacterial pathogen Campylobacter jejuni contains two subspecies: C. jejuni subsp. jejuni (Cjj and C. jejuni subsp. doylei (Cjd. Although Cjd strains are isolated infrequently in many parts of the world, they are obtained primarily from human clinical samples and result in an unusual clinical symptomatology in that, in addition to gastroenteritis, they are associated often with bacteremia. In this study, we describe a novel multiplex PCR method, based on the nitrate reductase (nap locus, that can be used to unambiguously subspeciate C. jejuni isolates. Results Internal and flanking napA and napB primer sets were designed, based on existing C. jejuni and Campylobacter coli genome sequences to create two multiplex PCR primer sets, nap mpx1 and nap mpx2. Genomic DNA from 161 C. jejuni subsp. jejuni (Cjj and 27 C. jejuni subsp. doylei (Cjd strains were amplified with these multiplex primer sets. The Cjd strains could be distinguished clearly from the Cjj strains using either nap mpx1 or mpx2. In addition, combination of either nap multiplex method with an existing lpxA speciation multiplex method resulted in the unambiguous and simultaneous speciation and subspeciation of the thermophilic Campylobacters. The Cjd nap amplicons were also sequenced: all Cjd strains tested contained identical 2761 bp deletions in napA and several Cjd strains contained deletions in napB. Conclusion The nap multiplex PCR primer sets are robust and give a 100% discrimination of C. jejuni subspecies. The ability to rapidly subspeciate C. jejuni as well as speciate thermophilic Campylobacter species, most of which are pathogenic in humans, in a single amplification will be of value to clinical laboratories in strain identification and the determination of the environmental source of campylobacterioses caused by Cjd. Finally, the sequences of the Cjd napA and napB loci suggest that Cjd strains arose from a common ancestor, providing clues as to

  13. Phage therapy reduces Campylobacter jejuni colonization in broilers

    NARCIS (Netherlands)

    Wagenaar, J.A.; Bergen, van M.A.P.; Mueller, M.A.; Wassenaar, T.M.; Carlton, R.M.

    2005-01-01

    The effect of phage therapy in the control of Campylobacter jejuni colonization in young broilers, either as a preventive or a therapeutic measure, was tested. A prevention group was infected with C. jejuni at day 4 of a 10-day phage treatment. A therapeutic group was phage treated for 6 days, start

  14. Host adaptation mechanisms and transcriptional regulation in Campylobacter jejuni

    NARCIS (Netherlands)

    van Mourik, A.

    2011-01-01

    Campylobacter jejuni is the most common cause of bacterial foodborne illness causing >100 million human cases each year worldwide. In contrast to other important enteropathogens such as Salmonella and Shigella spp., C. jejuni appears to lack a set of classical virulence traits, like adhesins, type I

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

    Science.gov (United States)

    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...

  16. In vitro susceptibilities of Campylobacter jejuni and Campylobacter coli to azithromycin and erythromycin.

    OpenAIRE

    Taylor, D E; Chang, N

    1991-01-01

    MICs of azithromycin and erythromycin for 20 Campylobacter coli and 20 Campylobacter jejuni strains were determined. The results demonstrated that, for Campylobacter species, all high-level erythromycin-resistant strains were also resistant to azithromycin and that azithromycin did not exhibit increased potency in comparison with that of erythromycin.

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

    Science.gov (United States)

    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.

  18. Cellular response of Campylobacter jejuni to trisodium phosphate

    DEFF Research Database (Denmark)

    Riedel, Charlotte Tandrup; Cohn, M. T.; Stabler, R. A.;

    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...

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

    Science.gov (United States)

    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.

  20. Pentavalent single-domain antibodies reduce Campylobacter jejuni motility and colonization in chickens.

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    Ali Riazi

    Full Text Available Campylobacter jejuni is the leading cause of bacterial foodborne illness in the world, with symptoms ranging from acute diarrhea to severe neurological disorders. Contaminated poultry meat is a major source of C. jejuni infection, and therefore, strategies to reduce this organism in poultry, are expected to reduce the incidence of Campylobacter-associated diseases. We have investigated whether oral administration of C. jejuni-specific single-domain antibodies would reduce bacterial colonization levels in chickens. Llama single-domain antibodies specific for C. jejuni were isolated from a phage display library generated from the heavy chain IgG variable domain repertoire of a llama immunized with C. jejuni flagella. Two flagella-specific single-domain antibodies were pentamerized to yield high avidity antibodies capable of multivalent binding to the target antigen. When administered orally to C. jejuni-infected two-day old chicks, the pentabodies significantly reduced C. jejuni colonization in the ceca. In vitro, the motility of the bacteria was also reduced in the presence of the flagella-specific pentabodies, suggesting the mechanism of action is through either direct interference with flagellar motility or antibody-mediated aggregation. Fluorescent microscopy and Western blot analyses revealed specific binding of the anti-flagella pentabodies to the C. jejuni flagellin.

  1. Quantitative Proteomics of Intracellular Campylobacter jejuni Reveals Metabolic Reprogramming.

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    Xiaoyun Liu

    Full Text Available 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.

  2. Exploring the chemotactic attraction of Campylobacter jejuni in chicken colonization

    DEFF Research Database (Denmark)

    Vegge, Christina Skovgaard; Brøndsted, Lone; Ingmer, Hanne

    Campylobacter jejuni is the primary food borne bacterial pathogen in the developed world. The most important reservoir for C. jejuni is the gut of chickens, which are colonized commensally and efficiently by this organism. Predominantly the mucus filled crypts of the lower gastrointestinal tract...... are found to be colonized by C. jejuni, and the bacteria are expected to be attracted to this particular environment by chemotaxis. In order to explore the role of chemotaxis in C. jejuni colonization we are construction deletion mutants in the putative chemoreceptors of the sequenced strain NCTC11168...

  3. Exploring the chemotatic attraction of Campylobacter jejuni in chicken colonization

    DEFF Research Database (Denmark)

    Vegge, Christina Skovgaard; Brøndsted, Lone; Ingmer, Hanne

    Campylobacter jejuni is the primary food borne bacterial pathogen in the developed world and the bacteria causes millions of gastroenteritis cases each year. The most important reservoir for C. jejuni is the gut of chickens, which are colonized commensally and efficiently by this organism....... Predominantly the mucus filled crypts of the lower gastrointestinal tract of chickens are found to be colonized by C. jejuni, and the bacteria are expected to be attracted to this particular environment by chemotaxis. From the full genome sequence of C. jejuni NCTC11168 several chemotactic proteins...

  4. Analysis of putative chemoreceptor proteins of Campylobacter jejuni

    DEFF Research Database (Denmark)

    Vegge, Christina Skovgaard; Brøndsted, Lone; Bang, Dang D.;

    Campylobacter jejuni is the primary food borne bacterial pathogen in the developed world. A very important reservoir for C. jejuni is the gut of chickens, which are colonized efficiently and commensally by this organism. Predominantly the mucus filled crypts of the lower gastrointestinal tract...... are found to be colonized by C. jejuni and the bacteria are expected to be attracted to this environment by chemotaxis. In order to explore the role of chemotaxis in C. jejuni colonization and to identify chemoreceptors with matching attractants and/or repellants we have constructed deletion mutants of five...

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

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    Kishan Kumar Nyati

    2013-01-01

    Full Text Available 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.

  6. Occurrence of Campylobacter jejuni and C. coli on broiler carcasses after chilling in southern Brazil

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    Gustavo Perdoncini

    2015-04-01

    Full Text Available Campylobacter jejuni and C. coli have been associated with gastrointestinal disorders in human beings, due mainly to the consumption of chicken meat. Despite control measures for reducing contamination by these bacteria, the detection of Campylobacter in carcasses after chilling remains high. A total of 105 carcasses were assessed by the horizontal detection method in five federally inspected slaughterhouses in southern Brazil in 2012 and in the first three months of 2013. Campylobacterwas isolated in 37.1% of the carcasses, of which 97.5% contained C. jejuni and 2.5% were infected by C. coli. The rate of positive carcasses across the slaughterhouses ranged from 0 to 71.4%. Determining the occurrence of Campylobacter among flocks is crucial for estimating the microbial load at specific points along the slaughtering process and for minimizing the risk of contamination of end products by Campylobacter.

  7. Evidence that certain clones of Campylobacter jejuni persist during successive broiler flock rotations

    DEFF Research Database (Denmark)

    Petersen, L.; Wedderkopp, A.

    2001-01-01

    Through the national surveillance program for Campylobacter spp,, nine broiler chicken farms that were infected with Campylobacter jejuni in at least five rotations in 1998 were identified. One additional farm, located at the island of Bornholm where divided slaughter is used extensively, was also...... selected. Twelve broiler houses located on 10 farms were included in the study. The C,jejuni isolates collected from the selected houses during the surveillance were typed using fla typing and macrorestriction profiling (MRP), and a subset of the isolates, representing each of the identified clones...... with recurrent Campylobacter problems. The MRPs of clones belonging to fla type 1/1 serotype O:2 isolated from persistently infected flocks shared a high percentage of bands compared to the remaining isolates, indicating that some clones that have the ability to cause persistent infections in broiler farms...

  8. The ability of Fla-typing schemes to discriminate between strains of Campylobacter jejuni

    DEFF Research Database (Denmark)

    Petersen, Line Hedegård; Newell, D.G.

    2001-01-01

    Aims: The aim of this investigation was to compare the usefulness of two previously published flagellin PCR-RFLP typing (Fla-typing) techniques for the subtyping of Campylobacter jejuni strains, in terms of ease of use and discriminatory power. Methods and Results: Six groups of isolates, which w...

  9. Molecular Evidence for Dissemination of Unique Campylobacter jejuni Clones in Curaçao, Netherlands Antilles

    NARCIS (Netherlands)

    Duim, B.; Godschalk, P.C.R.; Braak, N. van den; Dingle, K.E.; Dijkstra, J.R.; Leyde, E.; Plas, J. van der; Colles, F.M.; Endtz, H.P.; Wagenaar, J.A.; Maiden, M.C.J.; Belkum, A. van

    2003-01-01

    Campylobacter jejuni isolates (n = 234) associated with gastroenteritis and the Guillain-Barré syndrome (GBS) in the island of Curaçao, Netherlands Antilles, and collected from March 1999 to March 2000 were investigated by a range of molecular typing techniques. Data obtained by pulsed-field gel ele

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

    DEFF Research Database (Denmark)

    Birk, Tina; Grønlund, Anne Christine Jørgensen; Christensen, Bjarke Bak;

    2010-01-01

    The aim of this study was to determine whether marination of chicken meat in different food ingredients can be used to reduce populations of Campylobacter jejuni. C. jejuni strains were exposed to different organic acids (tartaric, acetic, lactic, malic, and citric acids) and food marinating...... ingredients at 4°C in broth and on chicken meat. The organic acids (0.5%) reduced populations of C. jejuni in broth (chicken juice and brain heart infusion broth) by 4 to 6 log units (after 24 h); tartaric acid was the most efficient treatment. Large strain variation was observed among 14 C. jejuni isolates...... inoculated in brain heart infusion broth containing 0.3% tartaric acid. On chicken meat medallions, reductions of C. jejuni were 0.5 to 2 log units when tartaric acid solutions (2, 4, 6, and 10%) were spread onto the meat. Analysis of acidic food ingredient (e.g., vinegar, lemon juice, pomegranate syrup...

  11. Importance of Campylobacter jejuni for Food Safety and Public Health

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    Omer Cakmak

    2010-04-01

    Full Text Available Campylobacter spp. are microorganisms that can be found in nature in the entire domestic and wild animal’s intestinal flora including the poultry and the sea animals. Campylobacter can better colonize in the poultry than the other animals. Campylobacter jejuni is an important pathogen among the thermophilic Campylobacter spp. whose growth temperature’s are different than the other Campylobacter spp. and can cause serious gastroenteritis in human beings which in some cases ended up with death. Human beings are generally infected with C. jejuni mainly because of the poultry meat and products and rarely because of the red meat which are contaminated during preparation and serving stages. Inadequate cooking, consumption of poorly chlorinated drinking water or unpasteurized milk are other infection sources of C. jejuni. Campylobacteriosis especially affect children under 5 years of age and reported to be a zoonotic illness that cause acute gastroenteritis in human. In many countries, food sourced C. jejuni infections were reported to occur more frequently than Salmonella spp. infections. In order to avoid Campylobacter infections, it is very important to enforce food security programmes and HACCP like systems during growth, slaughterhouses and point of sales stages. Also adequate cooking of the products, hygiene of the kitchen and personnel are important. [TAF Prev Med Bull 2010; 9(2.000: 157-166

  12. 鸡肉空肠弯曲杆菌的分离鉴定及耐药性分析%Isolation, identification and antimicrobial resistance analysis of Campylobacter jejuni isolates originated from chicken

    Institute of Scientific and Technical Information of China (English)

    韩新锋; 刘书亮; 张晓利; 陈荀; 侯小刚

    2012-01-01

    目的 分离和收集食源性空肠弯曲杆菌的优势流行株,了解市场上鸡肉中空肠弯曲杆菌的污染和药物敏感性情况.方法 根据GB/T4789.9-2003、SN0175-2010、ISO10272-1∶2006、FDA/BAM∶2001、USDA/FSIS∶1998等方法优化组合出一套对动物性食品源空肠弯曲杆菌有效的分离鉴定方法,从市场上鸡肉中分离疑似空肠弯曲杆菌,并进一步用生化试验及PCR方法进行准确鉴定.采用微量肉汤稀释法测定了所分离的空肠弯曲杆菌对22种常见抗菌药物(组合)的最低抑菌浓度值(MIC).结果与结论 根据培养特征、形态特征、生化试验和PCR结果准确鉴定出21株空肠弯曲杆菌,其污染率为11.5%.86%生化试验阳性的菌株经PCR鉴定为阳性.将空肠弯曲杆菌标准菌株和随机选出的分离株的16S rDNA扩增序列进行比对,二者同源率为100%.药敏试验表明,14.3%的分离菌株对氨苄西林具有耐药性;大多数菌株对头孢类抗生素MIC值相对较高;42.9%的菌株对萘啶酸具有耐药性;所有分离株对环丙沙星、四环素、红霉素、氯霉素、庆大霉素、链霉素不耐药.%According to the standards of GB/T4789. 9-2003, SN0175-2010, ISO10272-1:2006, FDA/BAM : 2001 and USDA/FSIS : 1998, a series of effective isolation and identification methods was developed for Campylobacter jejuni of chicken origin. By using these methods, suspected Campylobacter jejuni was firstly isolated from chicken in the market of Ya'an City, Sichuan Province, and then biochemical test and PCR method were used to further confirm the isolation strains. Combining the results of cultural characteristics, morphologic characteristics, biochemical test and PCR results, 21 isolates were accurately identified as Campylobacter jejuni, with which the contamination incidence was 11. 5% (21/183). The 86% of the strains identified by biochemical test were further confirmed by PCR. The 16S rDNA sequence comparison of the

  13. Characteristics of Lipo-Oligosaccharide Loci of Campylobacter jejuni Isolates Associated with Guillain-Barré Syndrome from Hebei, China

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    Jian-Zhong Zhang

    2010-03-01

    Full Text Available Ganglioside mimicry by C. jejuni lipo-oligosaccharides (LOS could induce the production of autoantibodies against gangliosides and the development of Guillain-Barré syndrome (GBS. The LOS biosynthesis region exhibits significant variation with different strains. Using PCR amplifications of genes from published LOS loci and sequencing the LOS biosynthesis loci, the eight GBS-associated C. jejuni strains from HeBei could be classified into four classes. The expression of sialylated LOS structures (class A or non-sialylated LOS structures(class F, H and P in the C. jejuni LOS is considered to be two different factors for the induction of GBS.

  14. Contamination analysis and PFGE typing of Campylobacter jejuni isolated from chicken production chain%鸡肉生产链中空肠弯曲杆菌的污染分析及PFGE分型研究

    Institute of Scientific and Technical Information of China (English)

    陈荀; 刘书亮; 吴聪明; 韩新锋; 陈栎娜; 陆思琴

    2012-01-01

    目的 了解鸡肉生产链中空肠弯曲杆菌的分布和传播情况.方法 参考GB/T4789.9-2008、SN0175-2010、ISO10272-1:2006、FDA/BAM:2001、USDA/FSIS:1998等方法组合出分离鸡源疑似空肠弯曲杆菌(Campylobacter jejuni,C.jejuni)的方法,采用16S rDNA和hipO基因的二重PCR方法对其准确鉴定;采用SmaⅠ对83株空肠弯曲杆菌进行PFGE分型.结果 从鸡肉生产链中采集样品519份,分离鉴定出空肠弯曲杆菌 180株,分离率为34.87%.其中养鸡场和屠宰场鸡粪样空肠弯曲杆菌分离率分别为74.29%和66.38%,屠宰场和市场的鸡肉样中空肠弯曲杆菌的分离率分别为17.33%和13.66%.83株空肠弯曲杆菌通过PFGE分型结果产生了45种不同的谱型.结论 四川省部分地区鸡肉生产链中空肠弯曲杆菌的污染较严重,空肠弯曲杆菌在鸡肉生产链中可能存在水平传播现象.%This study aims to understand the distribution and spread of Camp ylobacter jejuni in chicken production chain. According to the standards of GB/T4789. 9 2008, SN0175 2010, ISO10272 1:2006, FDA/BMA: 2001 and USDA/ FSIS; 1998, an effective isolation method was developed for detecting suspect Camp ylobacter jejuni contained chicken. The i solates were accurately identified by the duplex PCR based on the sequence of 16S rDNA and hipO gene. A total of 519 samples of the retail raw chicken and feces were examined for the presence of Campylobacter jejuni. From the chicken production chain, 180 Campylobacter jejuni strains were isolated, with an isolation rate of 34. 87%. The higher contamination rates were observed in feces from farms and slaughterhouse (74. 29% and 66. 38%, respectively). Whereas, the contamination rates in chicken from slaughterhouse and market were low (17. 33% and 13. 66% , respectively). The PFGE genotyping of 83 Campy lobacter jejuni isolates yielded 45 PFGE types. This demonstrated that the Campylobacter jejuni were widespread in chicken production chain, and the

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

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    Helen L Brown

    Full Text Available 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.

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

    Science.gov (United States)

    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.

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

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    Euna eOh

    2015-10-01

    Full Text Available 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.

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

    Science.gov (United States)

    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.

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

    Science.gov (United States)

    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

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

    Science.gov (United States)

    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.

  1. Campylobacter jejuni and Campylobacter coli in Children With Acute Diarrhea in Health Centers of Hamadan, Iran

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    Rastyani

    2015-11-01

    Full Text Available Background Enteritis caused by Campylobacter is considered as the most common acute bacterial diarrhea around the world. In most cases, infection occurs as a result of consuming contaminated water or food, especially raw meat of fowls. Objectives The purpose of the present study was to determine the prevalence and antibiotic resistance of campylobacter species among pediatrics of Hamadan city, Iran. Patients and Methods A total of 120 stool samples from children less than 10 years old were examined from January 2013 to December 2014 in Hamadan, Iran. The samples were incubated in Campy-Thio enrichment medium for 1 - 2 hours and then cultured on a specific medium; after that, the suspected colonies were analyzed for Campylobacter spp. identification by conventional tests. The identified species by biochemical methods were confirmed by polymerase chain reaction (PCR. Antimicrobial susceptibility testing was performed by disk agar diffusion (DAD method. Results Twelve (10% Campylobacter spp. from 120 stool samples were isolated including C. coli and C. jejuni. In the antibiotic susceptibility test, the most frequent resistance was observed to ciprofloxacin 8 (88.8%, followed by 7 (77.7% resistant strains to tetracycline, 7 (77.7% to erythromycin, 6 (66.6% to clindamycin, 5 (55.5% to meropenem, 4 (44.4% to gentamicin, 3 (33.3% to nalidixicacid and only 1 (11.1% to chloramphenicol. Conclusions Campylobacter is responsible for some important clinical problems such as enteritis and is also associated with meningitis and hemolytic-uremic syndrome. It is imperative to monitor the prevalence and antibiotic resistance of Campylobacter spp. as well as other the zoonotic bacteria.

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

    OpenAIRE

    Siringan, Patcharin; Connerton, Phillippa L.; Cummmings, 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 cy...

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

    Science.gov (United States)

    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.

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

    Science.gov (United States)

    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

  5. The occurrence and characterization of Campylobacter jejuni and C. coli in organic pigs and their outdoor environment

    DEFF Research Database (Denmark)

    Jensen, Annette Nygaard; Dalsgaard, Anders; Baggesen, Dorte Lau;

    2006-01-01

    to species by real-time PCR and sub-typed by serotyping (Penner) and pulse-field gel electrophorsis (PFGE) genotyping. All pigs excreted Campylobacter (10(3)-10(7) CFU g(-1) faeces) from the age of 8-13-weeks old. C jejuni was found in 29% of pigs in three consecutive trials and always in minority to C. coli...... (0.3-46%). C. jejuni and C coli were isolated from 10% and 29% of the environmental samples, respectively, while crow-birds and rats harboured C. jejuni. Individual pigs hosted several strains (up to nine serotypes). The paddock environment was contaminated with C. coli serotypes similar to pig...... isolates, while most of the C. jejuni serotypes differed. C. jejuni isolates of different origin comprised few similar serotypes, just one identical genotype was common between pigs, environment and birds. In conclusion, the occurrence of C jejuni varied considerably between the three groups of outdoor...

  6. Effect of Organic Acids and Marination Ingredients on the Survival of Campylobacter jejuni on Meat

    DEFF Research Database (Denmark)

    Birk, Tina; Grønlund, Anne Christine Jørgensen; Christensen, Bjarke Bak;

    2010-01-01

    The aim of this study was to determine whether marination of chicken meat in different food ingredients call be used to reduce populations of Campylobacter jejuni strains, were exposed to different organic acids (tartaric, acetic. lactic, malic, and citric acids) and food marinating ingredients...... at 4 degrees C in broth and on chicken meat. The organic acids (0.5%) reduced populations of C. jejuni broth (chicken juice and brain heart infusion broth) by 4 to 6 1011 units (after 24 h): tartaric acid was the most efficient treatment. Large strain variation was observed among 14 C. jejuni isolates...... inoculated in brain heart infusion broth containing 0.3% tartaric acid. On chicken meat medallions, reductions of C. jejuni were 0.5 to 2 log units when tartaric acid solutions (2, 4, 6, and 10%) were spread onto the meal. Analysis of acidic food ingredient (e.g., vinegar. lemon juice, pomegranate syrup...

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

    Science.gov (United States)

    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.

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

    Energy Technology Data Exchange (ETDEWEB)

    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.

  9. Colonization properties of Campylobacter jejuni in chickens

    OpenAIRE

    Pielsticker, C.; Glünder, G.; Rautenschlein, S.

    2012-01-01

    Campylobacter is the most common bacterial food-borne pathogen worldwide. Poultry and specifically chicken and raw chicken meat is the main source for human Campylobacter infection. Whilst being colonized by Campylobacter spp. chicken in contrast to human, do scarcely develop pathological lesions. The immune mechanisms controlling Campylobacter colonization and infection in chickens are still not clear. Previous studies and our investigations indicate that the ability to ...

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

    Science.gov (United States)

    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. PMID:26658311

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

    Science.gov (United States)

    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.

  12. Temperature-dependent phenotypic variation of Campylobacter jejuni lipooligosaccharides

    LENUS (Irish Health Repository)

    Semchenko, Evgeny A

    2010-11-30

    Abstract Background Campylobacter jejuni is a major bacterial cause of food-borne enteritis, and its lipooligosaccharide (LOS) plays an initiating role in the development of the autoimmune neuropathy, Guillain-Barré syndrome, by induction of anti-neural cross-reactive antibodies through ganglioside molecular mimicry. Results Herein we describe the existence and heterogeneity of multiple LOS forms in C. jejuni strains of human and chicken origin grown at 37°C and 42°C, respectively, as determined on sodium dodecyl sulphate-polyacrylamide electrophoresis gels with carbohydrate-specific silver staining and blotting with anti-ganglioside ligands, and confirmed by nuclear magnetic resonance (NMR) spectroscopy. The C. jejuni NCTC 11168 original isolate (11168-O) was compared to its genome-sequenced variant (11168-GS), and both were found to have a lower-Mr LOS form, which was different in size and structure to the previously characterized higher-Mr form bearing GM1 mimicry. The lower-Mr form production was found to be dependent on the growth temperature as the production of this form increased from ~5%, observed at 37°C to ~35% at 42°C. The structure of the lower-Mr form contained a β-D-Gal-(1→3)-β-D-GalNAc disaccharide moiety which is consistent with the termini of the GM1, asialo-GM1, GD1, GT1 and GQ1 gangliosides, however, it did not display GM1 mimicry as assessed in blotting studies but was shown in NMR to resemble asialo-GM1. The production of multiple LOS forms and lack of GM1 mimicry was not a result of phase variation in the genes tested of NCTC 11168 and was also observed in most of the human and chicken isolates of C. jejuni tested. Conclusion The presence of differing amounts of LOS forms at 37 and 42°C, and the variety of forms observed in different strains, indicate that LOS form variation may play a role in an adaptive mechanism or a stress response of the bacterium during the colonization of different hosts.

  13. Survival of Campylobacter jejuni in different gas mixtures

    DEFF Research Database (Denmark)

    Boysen, Louise; Knochel, Susanne; Rosenquist, Hanne

    2007-01-01

    Campylobacter jejuni in fresh chilled chicken meat is known to be a major risk factor for human gastrointestinal disease. In the present study, the survival under chilled conditions of different C. jejuni strains exposed to different gas mixtures usually used for gas packaging of food was examined....... jejuni were reduced by a minimum of 4.6 log(10) CFU mL(-1) over 21 days. When inoculated onto chicken fillets, the C. jejuni strains also died significantly faster in the oxygen-containing gas mixture, 70/30% O-2/CO2 (P <0.0001), reaching reductions of 2.0-2.6 log(10) CFU g(-1) after 8 days. In the gas...

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

    Science.gov (United States)

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

  15. Differential carbohydrate recognition by Campylobacter jejuni strain 11168: influences of temperature and growth conditions.

    Directory of Open Access Journals (Sweden)

    Christopher J Day

    Full Text Available The pathogenic clinical strain NCTC11168 was the first Campylobacter jejuni strain to be sequenced and has been a widely used laboratory model for studying C. jejuni pathogenesis. However, continuous passaging of C. jejuni NCTC11168 has been shown to dramatically affect its colonisation potential. Glycan array analysis was performed on C. jejuni NCTC11168 using the frequently passaged, non-colonising, genome sequenced (11168-GS and the infrequently passaged, original, virulent (11168-O isolates grown or maintained under various conditions. Glycan structures recognised and bound by C. jejuni included terminal mannose, N-acetylneuraminic acid, galactose and fucose. Significantly, it was found that only when challenged with normal oxygen at room temperature did 11168-O consistently bind to sialic acid or terminal mannose structures, while 11168-GS bound these structures regardless of growth/maintenance conditions. Further, binding of un-capped galactose and fucosylated structures was significantly reduced when C. jejuni was maintained at 25 degrees C under atmospheric oxygen conditions. These binding differences identified through glycan array analysis were confirmed by the ability of specific lectins to competitively inhibit the adherence of C. jejuni to a Caco-2 intestinal cell line. Our data suggests that the binding of mannose and/or N-acetylneuraminic acid may provide the initial interactions important for colonisation following environmental exposure.

  16. The influence of age on Campylobacter jejuni infection in chicken.

    Science.gov (United States)

    Han, Zifeng; Pielsticker, Colin; Gerzova, Lenka; Rychlik, Ivan; Rautenschlein, Silke

    2016-09-01

    Campylobacter jejuni (C. jejuni)-host-interaction may be affected by the maturation stage of the chicken's immune system and the developing gut microbiota composition. We compared these parameters between birds C. jejuni-inoculated at day one, 10, 22 and 31 post hatch. The highest C. jejuni-colonization rate and numbers of colony forming units (CFU) were detected in caecal content of day-one-inoculated birds while the lowest was detected in 22-days-old birds. The low bacterial colonization of 22-days-old chickens correlated with the most prominent immune reactions in this age group in comparison to other age groups. Age and C. jejuni-inoculation had a significant effect on lymphocyte numbers and cytokine expression levels in caecum as well as on gut flora composition. Overall, the immune response to C. jejuni is significantly influenced by the age of the infected chickens leading to differences in C. jejuni-colonization pattern between age goups. PMID:27131855

  17. Genomic characterization of Campylobacter jejuni strain M1.

    Directory of Open Access Journals (Sweden)

    Carsten Friis

    Full Text Available Campylobacter jejuni strain M1 (laboratory designation 99/308 is a rarely documented case of direct transmission of C. jejuni from chicken to a person, resulting in enteritis. We have sequenced the genome of C. jejuni strain M1, and compared this to 12 other C. jejuni sequenced genomes currently publicly available. Compared to these, M1 is closest to strain 81116. Based on the 13 genome sequences, we have identified the C. jejuni pan-genome, as well as the core genome, the auxiliary genes, and genes unique between strains M1 and 81116. The pan-genome contains 2,427 gene families, whilst the core genome comprised 1,295 gene families, or about two-thirds of the gene content of the average of the sequenced C. jejuni genomes. Various comparison and visualization tools were applied to the 13 C. jejuni genome sequences, including a species pan- and core genome plot, a BLAST Matrix and a BLAST Atlas. Trees based on 16S rRNA sequences and on the total gene families in each genome are presented. The findings are discussed in the background of the proven virulence potential of M1.

  18. Campylobacter jejuni, Campylobacter coli, and cytolethal distending toxin (CDT) genes in common teals (Anas crecca)

    OpenAIRE

    Gargiulo, Antonio; Sensale, Mariangela; Marzocco, Laura; Fioretti, Alessandro; Menna, Lucia F.; Dipineto, Ludovico

    2011-01-01

    Abstarct To evaluate the presence of Campylobacter spp. and related cdt genes, cloacal swabs were collected from 70 common teals (Anas crecca) and analyzed by culture methods and polymerase chain reaction. In addition, C. jejuni were examined also for the presence of wlaN gene. This is believed to be the first report of Campylobacter spp. in common teal and our results confirm the very common occurrence of C. jejuni (n=40) and C. coli (n=13) in waterfowls. Furthermore, the cdt gene...

  19. Campylobacter jejuni and Campylobacter coli in wild birds on Danish livestock farms

    DEFF Research Database (Denmark)

    Hald, Birthe; Skov, Marianne Nielsine; Nielsen, Eva Møller;

    2016-01-01

    , fat score, gender, and migration range were not found to be associated with Campylobacter spp. carriage. A correlation was found between the prevalence (%) of C. jejuni in wild birds and the proportions (%) of C. jejuni in both manure on cattle farms (R-2 = 0.92) and poultry farms (R-2 = 0....... The farm environment provides attractive foraging and breeding habitats for some bird species reported to carry thermophilic Campylobacter spp. We investigated the Campylobacter spp. carriage rates in 52 wild bird species present on 12 Danish farms, sampled during a winter and a summer season, in order...... to study the factors influencing the prevalence in wild birds according to their ecological guild. In total, 1607 individual wild bird cloacal swab samples and 386 livestock manure samples were cultured for Campylobacter spp. according to the Nordic Committee on Food Analysis method NMKL 119.Results...

  20. Comparison of Disk Diffusion and Agar Dilution Methods for Erythromycin and Ciprofloxacin Susceptibility Testing of Campylobacter jejuni subsp. jejuni▿

    OpenAIRE

    Gaudreau, Christiane; Girouard, Yves; Ringuette, Louise; Tsimiklis, Catherine

    2007-01-01

    Disk diffusion was a reliable, easy, and inexpensive method for testing the susceptibility of Campylobacter jejuni to erythromycin (215 susceptible and 45 resistant isolates) and to ciprofloxacin (154 susceptible, two intermediate, and 124 resistant isolates) using, respectively, an erythromycin disk and ciprofloxacin and nalidixic acid disks.

  1. Evaluation of modified CCD agar for isolation of Campylobacter jejuni%空肠弯曲菌分离培养基改良CCD琼脂的性能测试研究

    Institute of Scientific and Technical Information of China (English)

    何天文; 陈佐威; 苏丽春; 蔡芷荷; 吴清平

    2012-01-01

    目的:比较空肠弯曲菌(Campylobacter jejuni)在国内外四个厂家的改良CCD琼脂上生长的灵敏度及回收率.方法:用哥伦比亚血平板作为参照培养基,根据ISO/TS11133.2-2003标准检验方法对空肠弯曲菌分离培养基进行性能测试.结果:样品本底测试显示,在四个厂家培养基中测试的样品均没有检出可疑的空肠弯曲菌,但厂家②对杂菌的抑制能力较差;各厂家的的改良CCD琼脂灵敏度均可达到10 CFU以内;环凯、厂家①、厂家②、OXOID四个厂家的改良CCD琼脂的回收率分别为83.4%、46.4%、32.4%、89.4%,均能达到ISO/TS11133.2-2003标准检验方法的要求.结论:环凯和OXOID的改良CCD琼脂分离空肠弯曲菌的效果无明显差异,但均优于国内其他两个厂家(P<0.05).%Objective:To compare the sensitivity and recovery rates of four media ( modified CCD agar) for isolation of Campylobacter jejuni. Methods: Using the Columbia blood agar as the reference medium, the performance of four different media for isolation of C. jejuni was evaluated according to ISO/TS11133. 2 -2003 in this study. Results; There were no C. jejuni in the samples;The sensitivity of each modified CCD agar was all within 10 CFU. The recovery rates of modified CCD agar from Huankai, brand No. 1, brand No. 2 and Oxiod were 83. 4% ,46. 4% , 32.4% and 89. 4% respectively, all could meet the requirement of ISO/TS11133. 2 - 2003. Conclusion; There were no obvious differences between Huankai and Oxiod modified CCD agar for isolating C. jejuni, and they were better than other two.

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

    Science.gov (United States)

    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

  3. Computer-assisted analysis and epidemiological value of genotyping methods for Campylobacter jejuni and Campylobacter coli

    NARCIS (Netherlands)

    Boer, P. de; Duim, B.; Rigter, A.; Plas, J. van der; Jacobs-Reitsma, W.F.; Wagenaar, J.A.

    2000-01-01

    For epidemiological tracing of the thermotolerant Campylobacter species C. jejuni and C. coli, reliable and highly discriminatory typing techniques are necessary. In this study the genotyping techniques of flagellin typing (flaA typing), pulsed-field gel electrophoresis (PFGE), automated ribotyping,

  4. Chemical Decontamination of Campylobacter jejuni on Chicken Skin and Meat

    DEFF Research Database (Denmark)

    Riedel, Charlotte Tandrup; Brøndsted, Lone; Rosenquist, Hanne;

    2009-01-01

    This study evaluated the effectiveness of 11 chemical compounds to reduce Campylobacter jejuni on chicken skin and meat samples dipped in chemical solutions. Treatment of skin samples for 1 min using tartaric acid (2%) and caprylic acid sodium salt (5%) caused reductions of C. jejuni NCTC11168......, which were not significantly different from the reduction obtained by sterile water (0.95 log). Statistically larger reductions (1.57 to 3.81 log) were caused by formic acid (2%), lactic acid (2.5%), trisodium phosphate (10%), capric acid sodium salt (5%), grapefruit seed extract (1......, sterile water and lactic acid caused considerably larger reductions on skin than on meat, whereas the opposite was seen for caprylic acid sodium salt. In conclusion, this study has identified chemicals with substantial reduction effects on C. jejuni. The analysis has further emphasized that treatment time...

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

    Energy Technology Data Exchange (ETDEWEB)

    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.

  6. Chemical Decontamination of Campylobacter jejuni on Chicken Skin and Meat

    DEFF Research Database (Denmark)

    Riedel, Charlotte Tandrup; Brøndsted, Lone; Rosenquist, Hanne;

    2009-01-01

    This study evaluated the effectiveness of 11 chemical compounds to reduce Campylobacter jejuni on chicken skin and meat samples dipped in chemical solutions. Treatment of skin samples for 1 min using tartaric acid (2%) and caprylic acid sodium salt (5%) caused reductions of C. jejuni NCTC11168...... chlorhexidine diacetate salt hydrate (1%). The most effective compounds were cetylpyridinium chloride (0.5%) and benzalkonium chloride (1%) (>4.2 log). However, when these treated samples were stored for 24 h at 5°C, cetylpyridinium chloride, benzalkonium chloride, and grapefruit seed extract were less...... effective, indicating that some cells may recover after a 1-min treatment with these chemicals. An increase in treatment time to 15 min resulted in higher effectiveness of trisodium phosphate and formic acid. Interestingly, when reduction of the C. jejuni population was compared on chicken skin and meat...

  7. Differentiation of the virulence potential of Campylobacter jejuni strains by use of gene transcription analysis and a caco-2 assay

    DEFF Research Database (Denmark)

    Poli, Vanessa Fadanelli Schoenardie; Thorsen, Line; Olesen, Inger;

    2012-01-01

    Campylobacter jejuni is the leading cause of bacterial diarrheal disease in humans, and contaminated poultry and poultry products are recognized as the main vehicle of infection. Despite the significance of C. jejuni as a foodborne pathogen, little is known about its response to stress, and...... expression of C. jejuni. The clinical strains appeared to be more virulent than the chicken isolate as measured by the Caco-2 invasion assay which could be due to differences in CipA functionality. The RT-qPCR analysis and Caco-2 assay showed to be useful tools for differentiating virulence potentials...

  8. Influxed insects as Vectors for Campylobacter jejuni and Campylobacter coll in Danish Broiler Houses

    DEFF Research Database (Denmark)

    Hald, Birthe; Skovgård, Henrik; Pedersen, Karl;

    2008-01-01

    houses was estimated by trapping of insects (n = 5,936) in ventilation vents. In total, 31 flies (28 of which were of the Muscidae family) caught in farm surroundings were Campylobacter spp.-positive (C. jejuni, n = 7; C. coli, n = 23; other Campylobacter spp., n = 1). Musca domestica (L) (house fly...... caesar (L) (green bottle fly) of the Calliphoridae family and 2 flies of unidentified species were also positive. The prevalence of Campylobacter spp.-positive flies varied from 0.0 in April to a peak of 16.3% in July and decreasing to 2.0% in October on a farm with pig production. On 4 broiler farms.......0078) and with the outdoor temperature (P houses constitutes a considerable risk for infection of broilers with C. jejuni and C. coli....

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

    Directory of Open Access Journals (Sweden)

    Petra Griekspoor

    2015-12-01

    Full Text Available 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.

  10. Status of vaccine research and development for Campylobacter jejuni.

    Science.gov (United States)

    Riddle, Mark S; Guerry, Patricia

    2016-06-01

    Campylobacter jejuni is one of the leading causes of bacterial diarrhea worldwide and is associated with a number of sequelae, including Guillain-Barre Syndrome, reactive arthritis, irritable bowel syndrome and growth stunting/malnutrition. Vaccine development against C. jejuni is complicated by its antigenic diversity, a lack of small animal models, and a poor understanding of the bacterium's pathogenesis. Vaccine approaches have been limited to recombinant proteins, none of which have advanced beyond Phase I testing. Genomic analyses have revealed the presence of a polysaccharide capsule on C. jejuni. Given the success of capsule-conjugate vaccines for other mucosal pathogens of global importance, efforts to evaluate this established approach for C. jejuni are also being pursued. A prototypical capsule-conjugate vaccine has demonstrated efficacy against diarrheal disease in non-human primates and is currently in Phase I testing. In addition to proof of concept studies, more data on the global prevalence of capsular types, and a better understanding of the acute and chronic consequences of C. jejuni are needed to inform investments for a globally relevant vaccine. PMID:26973064

  11. Campylobacter jejuni motility is required for infection of the flagellotropic bacteriophage F341

    DEFF Research Database (Denmark)

    Baldvinsson, Signe Berg; Sørensen, Martine Camilla Holst; Vegge, Christina Skovgaard;

    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...

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

    Science.gov (United States)

    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...

  13. Comparison of genotypes and serotypes of Campylobacter jejuni isolated from Danish wild mammals and birds and from broiler flocks and humans

    DEFF Research Database (Denmark)

    Petersen, L.; Nielsen, E.M.; Engberg, J.;

    2001-01-01

    (MRPs) of C. jejuni isolates from different sources. The serotype distribution in wildlife was significantly different from the known distributions in broilers and humans. Considerable sero- and genotype diversity was found within the wildlife collection, although two major groups of isolates within...... serotype O:12 and the O:4 complex were found. Common clonal lines among wildlife, chicken, and/or human isolates were identified within serotype O:12 and the O:4 complex. However, MRPs of O:12 and O:38 strains isolated from wildlife and other sources indicated that some clonal lines propagated in a wide...

  14. Biological roles of the O-methyl phosphoramidate capsule modification in Campylobacter jejuni.

    Directory of Open Access Journals (Sweden)

    Lieke B van Alphen

    Full Text Available 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.

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

    Science.gov (United States)

    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

  16. Targeting motility properties of bacteria in the development of probiotic cultures against Campylobacter jejuni in broiler chickens.

    Science.gov (United States)

    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 (pCampylobacter 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.

  17. Campylobacter jejuni colonization in wild birds: results from an infection experiment.

    Directory of Open Access Journals (Sweden)

    Jonas Waldenström

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

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

    Science.gov (United States)

    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.

  19. Biotyping of thermophilic Campylobacter spp. isolated from poultry in and around Anand city, Gujarat, India

    Directory of Open Access Journals (Sweden)

    R. S. Tayde

    2014-05-01

    Full Text Available Aim: To study the prevalence of different biotypes of thermophilic Campylobacter spp. in the study area. Materials and Methods: A total of 150 samples comprising 90 chicken and 60 caecal content were collected from retail meat market and processed for isolation of Campylobacter spp. 52 Campylobacter isolates obtained from raw poultry meat (6 and caecal content (46 were subjected to biotyping using Lior's biotyping scheme. Results: Among the 52 Campylobacter isolates studied, 60.46 % isolates were identified as Campylobacter jejuni Biotype I and 39.53% were C. jejuni Biotype II, whereas 83.33 % were C. coli Biotype I and 16.66 % C. coli Biotype II. No other biotypes were identified. Conclusions: The present study revealed that C. jejuni Biotype I was more prevalent than Biotype II whereas in case of C. coli, Biotype I was more prevalent than Biotype II providing basis for further epidemiological study.

  20. Survival of Campylobacter jejuni in biofilms after chlorine treatment

    Directory of Open Access Journals (Sweden)

    Kunyaboon, S

    2006-09-01

    Full Text Available Survival of C. jejuni in biofilms isolated from two chicken houses in Thailand (FBRL-C04, FBRLB05 and FBRL-B06 after chlorine treatment was studied. Biofilm cultures were grown on stainless steel surface in 50% trypticase soy broth for 3 days, subsequently C. jejuni cells were allowed to attach to these biofilms for 4 h at 25ºC. Sodium hypochlorite was used to prepare sanitizing solution with active chlorine of 15 ppm and 25 ppm. Stainless steel coupons containing C. jejuni with and without biofilms were treated with chlorine for 30 sec and neutralized with 0.05% sodium thiosulfate. At both concentrations, C. jejuni were inactivated to lower than 1 log10CFU/cm2 when initial attachment load was approximately 4 log10CFU/cm2. However, C. jejuni in all samples treated with 15 ppm active chlorine were recovered in enrichment media. When treated with the higher concentration of chlorine, 25 ppm, C. jejuni in biofilm of FBRL-C04 (5/9, FBRL-B06 (1/9 and biofilm-free surface (1/9 could also be recovered. This indicates that chlorine treatment at 15 and 25 ppm could not completely inactivate C. jejuni attached to biofilms and biofilm-free surfaces. Biofilm of FBRL-C04 enhanced the survival of C. jejuni after chlorine treatment at 25 ppm although biofilm initial attachment as determined by plate count method was similar to that of other biofilms. Attachment load of viable biofilm cells may not contribute to enhanced survival of C. jejuni in chlorine treatment.

  1. Cj1411c GENE OF CAMPYLOBACTER JEJUNI 11168 ENCODES FOR A CYTOCHROME P450 INVOLVED IN BACTERIAL CAPSULE SUGAR METABOLISM

    Directory of Open Access Journals (Sweden)

    N. CORCIONIVOSCHI

    2013-12-01

    Full Text Available After isolation in 1970s, Campylobacter jejuni become the most commonlyrecognized cause of bacterial gastroenteritis in man. In animals is frequently foundin bovines on ovines. Publishing of the genome sequence of Campylobacter jejuni11168 (Parkhill, 2000 revealed the presence of only one cytochrome P450 in anoperon involved in sugar and cell surface biosynthesis. The gene name is Cj1411c, is1359 bp long and encodes 453 aa. The sequence is strictly conserved inCampylobacter jejuni RM221. Similarities with two cytochrome P450s, one formSilicobacter sp. and one form Poloromonas sp., were identified. These two enzymesare known to be involved in ascorbate and aldarate metabolism. The recombinantconstruct allowed the expression of active P450 enzyme with a 450 nm peak whenbinds CO. The protein was purified in proportion of ~ 70 %. By deleting the P450gene from the Campylobacter jejuni 11168 genome clear changes in cellmorphology were identified cells becoming wider and shorter. The capsular sugarprofile of the NCI strain reveals the presence of arabinose which was not found inthe wild type strain. The arabinose was identified by both High Performance LiquidChromatography (HPLC and Nuclear Magnetic Resonance (NMR.

  2. Cj1411c GENE OF CAMPYLOBACTER JEJUNI 11168 ENCODES FOR A CYTOCHROME P450 INVOLVED IN BACTERIAL CAPSULE SUGAR METABOLISM

    Directory of Open Access Journals (Sweden)

    CORCIONIVOSCHI N.

    2007-01-01

    Full Text Available After isolation in 1970s, Campylobacter jejuni become the most commonlyrecognized cause of bacterial gastroenteritis in man. In animals is frequently foundin bovines on ovines. Publishing of the genome sequence of Campylobacter jejuni11168 (Parkhill, 2000 revealed the presence of only one cytochrome P450 in anoperon involved in sugar and cell surface biosynthesis. The gene name is Cj1411c, is1359 bp long and encodes 453 aa. The sequence is strictly conserved inCampylobacter jejuni RM221. Similarities with two cytochrome P450s, one formSilicobacter sp. and one form Poloromonas sp., were identified. These two enzymesare known to be involved in ascorbate and aldarate metabolism. The recombinantconstruct allowed the expression of active P450 enzyme with a 450 nm peak whenbinds CO. The protein was purified in proportion of ~ 70 %. By deleting the P450gene from the Campylobacter jejuni 11168 genome clear changes in cellmorphology were identified cells becoming wider and shorter. The capsular sugarprofile of the NCI strain reveals the presence of arabinose which was not found inthe wild type strain. The arabinose was identified by both High Performance LiquidChromatography (HPLC and Nuclear Magnetic Resonance (NMR.

  3. Comparative genomic analysis of Campylobacter jejuni associated with Guillain-Barré and Miller Fisher syndromes: neuropathogenic and enteritis-associated isolates can share high levels of genomic similarity

    Directory of Open Access Journals (Sweden)

    Endtz Hubert P

    2007-10-01

    Full Text Available Abstract Background Campylobacter jejuni infection represents the most frequent antecedent infection triggering the onset of the neuropathic disorders Guillain-Barré syndrome (GBS and Miller Fisher syndrome (MFS. Although sialylated ganglioside-mimicking lipo-oligosaccharide (LOS structures are the strongest neuropathogenic determinants in C. jejuni, they do not appear to be the only requirement for a neuropathic outcome since strains capable of their production have been isolated from patients with uncomplicated cases of enteritis. Consequently, other pathogen and/or host-related factors contribute to the onset of neurological complications. We have used comparative genomic hybridization to perform a detailed genomic comparison of strains isolated from GBS/MFS and enteritis-only patients. Our dataset, in which the gene conservation profile for 1712 genes was assayed in 102 strains, including 56 neuropathogenic isolates, represents the largest systematic search for C. jejuni factors associated with GBS/MFS to date and has allowed us to analyze the genetic background of neuropathogenic C. jejuni strains with an unprecedented level of resolution. Results The majority of GBS/MFS strains can be assigned to one of six major lineages, suggesting that several genetic backgrounds can result in a neuropathogenic phenotype. A statistical analysis of gene conservation rates revealed that although genes involved in the sialylation of LOS structures were significantly associated with neuropathogenic strains, still many enteritis-control strains both bear these genes and share remarkable levels of genomic similarity with their neuropathogenic counterparts. Two capsule biosynthesis genes (Cj1421c and Cj1428c showed higher conservation rates among neuropathogenic strains compared to enteritis-control strains. Any potential involvement of these genes in neuropathogenesis must be assessed. A single gene (HS:3 Cj1135 had a higher conservation rate among enteritis

  4. Identification of immunogenic and virulence-associated Campylobacter jejuni proteins

    DEFF Research Database (Denmark)

    Nielsen, Lene Nørby; Luijkx, Thomas A.; Vegge, Christina Skovgaard;

    2012-01-01

    With the aim of identifying proteins important for host interaction and virulence, we have screened an expression library of NCTC 11168 Campylobacter jejuni genes for highly immunogenic proteins. A commercial C. jejuni open reading frame (ORF) library consisting of more than 1,600 genes was trans......With the aim of identifying proteins important for host interaction and virulence, we have screened an expression library of NCTC 11168 Campylobacter jejuni genes for highly immunogenic proteins. A commercial C. jejuni open reading frame (ORF) library consisting of more than 1,600 genes...

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

    Science.gov (United States)

    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

  6. Isolation and Characterization of Campylobacter Strains from Diarrheal Patients in Central and Suburban Bangkok, Thailand.

    Science.gov (United States)

    Samosornsuk, Worada; Asakura, Masahiro; Yoshida, Emi; Taguchi, Takashi; Eampokalap, Bunchuay; Chaicumpa, Wanpen; Yamasaki, Shinji

    2015-01-01

    Campylobacter-induced diarrhea is increasingly recognized worldwide. However, little information is available regarding the Campylobacter strains associated with diarrheal patients in Thailand. In this study, we attempted to isolate Campylobacter strains from diarrheal patients in Thailand and to characterize the species using a cytolethal distending toxin (cdt) gene-based C. jejuni, C. coli, and C. fetus-specific multiplex PCR assay. Campylobacter species were also confirmed using 16S rRNA gene sequencing and hipO gene detection. From 2,500 diarrheal stool specimens, 76 Campylobacter-like organisms were isolated and identified via conventional culture methods. Among these 76 organisms, 73 were identified as Campylobacter species (43 C. jejuni, 29 C. coli, and 1 C. fetus) via multiplex PCR, whereas 3 remained unidentified. Two Campylobacter-like organisms yielded 2 amplicons corresponding to cdt genes from C. jejuni and C. coli. Subsequently, C. jejuni and C. coli were reisolated from each sample. The third isolate was identified as C. hyointestinalis via 16S rRNA gene sequencing. To our knowledge, this is the first report on the isolation of C. hyointestinalis from a diarrheal patient in Thailand. These data indicate that C. jejuni (58%) and C. coli (40%) are prevalent among diarrheal patients in Thailand.

  7. Prevalence of Campylobacter, Arcobacter, Helicobacter, and Sutterella spp. in human fecal samples as estimated by a reevaluation of isolation methods for Campylobacters

    DEFF Research Database (Denmark)

    Engberg, J.; On, Stephen L.W.; Harrington, C.S.;

    2000-01-01

    CCDA recovered significantly more thermophilic Campylobacter spp. than Skirrow's medium (P = 0.0034). No significant difference between Skirrow's medium and CAT agar was observed in this study. Another six taxa were identified, namely, Campylobacter concisus, Campylobacter curvus-like bacteria, Arcobacter......The aims of this study were to investigate the prevalence of campylobacteria including Campylobacter jejuni subsp. jejuni (C. jejuni) and Campylobacter coli in human clinical samples and in samples from healthy individuals and to reevaluate the efficacies of conventional selective methods...... for isolation of Campylobacter spp. Two charcoal-based selective media, modified charcoal cefoperazone deoxycholate agar (mCCDA) and cefoperazone-amphotericin-teicoplanin (CAT) agar, were compared with Skirrow's blood-based medium and with a filter method (filter) applied to a yeast-enriched blood agar. A total...

  8. Cloning and alignment of WaaF gene of Campylobacter jejuni Lulei

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    XING Cong-cong

    2012-04-01

    Full Text Available Objective To clone the WaaF gene of Campylobacter jejuni, and analyse its relationship with WaaF genetic evolution. Methods Amplified WaaF gene of Campylobacter jejuni Lulei by PCR, and constructed pGEM-T-WaaF cloning plasmid. Downloaded five WaaF associated with Guillain-Barré syndrome (GBS and one WaaF not associated with GBS, and then constructed phylogenetic tree. Results pGEM-T-WaaF cloning plasmid was constructed successfully. WaaF presented cluster phenomenon in Campylobacter jejuni associated with GBS. Conclusion WaaF gene of Campylobacter jejuni Lulei is the fragment of 807 bp, and has the nearest relationship with the genetic evolution of Lichang.

  9. Development of a Chemically Defined Medium suitable for [35S]-methionine labeling of Campylobacter jejuni

    DEFF Research Database (Denmark)

    Birk, Tina; Rosenquist, Hanne; Knøchel, Susanne

    Campylobacter jejuni is a micro-aerobic foodborne pahtogenic bacteria generally regarded as one of the major causes of gastroenteritis in humans worlwoide. Consumption and handling of fresh poultry meat are considered the primary sources of campylobacteriosis in Denmark.......Campylobacter jejuni is a micro-aerobic foodborne pahtogenic bacteria generally regarded as one of the major causes of gastroenteritis in humans worlwoide. Consumption and handling of fresh poultry meat are considered the primary sources of campylobacteriosis in Denmark....

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

    Science.gov (United States)

    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

  11. In vitro susceptibility of Campylobacter jejuni from Kuwait to tigecycline & other antimicrobial agents

    Directory of Open Access Journals (Sweden)

    M John Albert

    2013-01-01

    Full Text Available Background & objectives: There is an increasing frequency of resistance of Campylobacter jejuni to antimicrobial agents making treatment difficult. In this study, the in vitro susceptibility of C. jejuni isolates collected over an eight year period was tested against tigecycline, a glycylcycline, the previously tested antimicrobial agents in Kuwait, ciprofloxacin, erythromycin and tetracycline, and other antimicrobial agents not previously tested in Kuwait, amoxicillin-clavulanic acid, gentamicin, imipenem and meropenem. Methods: A total of 97 C. jejuni isolates from diarrhoeal stools of Kuwaiti patients during 2002-2010 were studied for susceptibility to the above antimicrobial agents by E test. Results: Erythromycin resistance increased from 5.0 per cent in 2002-2003 to 13.8 per cent in 2007-2010. The figures for ciprofloxacin resistance for the same periods were 53 and 65.5 per cent, respectively. Tetracycline resistance increased from 40.0 per cent in 2003-2006 to 62.1 per cent in 2007-2010 (P=0.05. However, all isolates were uniformly susceptible to tigecycline and other antimicrobial agents. Interpretation & conclusions: There was a progressive increase in the prevalence of resistance to ciprofloxacin, erythromycin and tetracycline. As all isolates were uniformly susceptible to tigecycline, this antimicrobial agent can be considered as a potential candidate for treatment in clinical studies.

  12. Antigenic protein synthesis of Campylobacter jejuni in contact with chicken cells

    DEFF Research Database (Denmark)

    Vegge, Christina Skovgaard; Bang, Dang D.; Li, Yiping;

    Campylobacter jejuni is the primary food borne bacterial pathogen in the developed world causing millions of gastroenteritis cases each year. C. jejuni is a Gram negative, spiral-shaped, highly motile bacterium with very restricted growth requirements, and it appears to be adapted to the environm......Campylobacter jejuni is the primary food borne bacterial pathogen in the developed world causing millions of gastroenteritis cases each year. C. jejuni is a Gram negative, spiral-shaped, highly motile bacterium with very restricted growth requirements, and it appears to be adapted to the...... environment of the avian gastrointestinal tract. Consequently, the most important reservoir for C. jejuni is the gut of chickens, which are colonized efficiently without causing disease in the birds. Upon co-cultivation with mammalian cells, C. jejuni secrete specific Cia proteins, which are required for...

  13. Antigenic protein synthesis of Campylobacter jejuni in contact with chicken cells

    DEFF Research Database (Denmark)

    Vegge, Christina Skovgaard; Bang, Dang D.; Li, Yiping;

    Campylobacter jejuni is the primary food borne bacterial pathogen in the developed world causing millions of gastroenteritis cases each year. C. jejuni is a Gram negative, spiral-shaped, highly motile bacterium with very restricted growth requirements, and it appears to be adapted to the environm......Campylobacter jejuni is the primary food borne bacterial pathogen in the developed world causing millions of gastroenteritis cases each year. C. jejuni is a Gram negative, spiral-shaped, highly motile bacterium with very restricted growth requirements, and it appears to be adapted to the...... environment of the avian gastrointestinal tract. Consequently, the most important reservoir for C. jejuni is the gut of chickens, which are colonized efficiently without causing disease in the birds. Upon co-cultivation with mammalian cells, C. jejuni secrete specific Cia proteins, which are required for...

  14. Detection of a putative virulence cadF gene of Campylobacter jejuni obtained from different sources using a microfabricated PCR chip

    DEFF Research Database (Denmark)

    Poulsen, Claus Riber; El-Ali, Jamil; Perch-Nielsen, Ivan R.;

    2005-01-01

    A microfabricated polymerase chain reaction (PCR) chip made of epoxy-based photoresist (SU-8) was recently designed and developed. In this study, we tested whether the PCR chip could be used for rapid detection of a potential virulence determinant, the cadF gene of Campylobacter jejuni. PCR...... was performed using published PCR conditions and primers for the C. jejuni cadF gene. DNA isolated from a C. jejuni reference strain CCUG 11284, C. jejuni isolates obtained from different sources (chicken and human), and Campylobacter whole cells were used as templates in the PCR tests. Conventional PCR in tube...... was used as the control. After optimization of the PCR chip, PCR positives on the chip were obtained from 91.0% (10/11) of the tested chips. A fast transition time was achieved with the PCR chip, and therefore a faster cycling time and a shorter PCR program were obtained. Using the PCR chip, the cadF gene...

  15. Comparative genotyping of Campylobacter jejuni strains from patients with Guillain-Barré syndrome

    NARCIS (Netherlands)

    Z. Islam (Zhahirul); A.F. van Belkum (Alex); J.A. Wagenaar (Jaap); A.J. Cody (Alison); A.G. de Boer (Albert); H. Tabor (Helen); B.C. Jacobs (Bart); K.A. Talukder (Kaisar); H.P. Endtz (Hubert)

    2009-01-01

    textabstractBackground: Campylobacter jejuni is a common cause of acute gastroenteritis and is associated with post-infectious neuropathies such as the Guillain-Barré syndrome (GBS) and the Miller Fisher syndrome (MFS). We here present comparative genotyping of 49 C. jejuni strains from Bangladesh t

  16. QUANTIFICATIVE ANALYSIS OF VIABLE, STRESSED AND DEAD CELLS OF CAMPYLOBACTER JEJUNI STRAIN 81-176

    Science.gov (United States)

    Campylobacter jejuni is an important foodborne gastrointestinal pathogen. Research has shown that changes in culturability, cell morphology, and viability occur when C. jejuni cells are subjected to stresses such as low nutrient availability, entry into stationary phase, or low CO2/high O2 condition...

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

    Science.gov (United States)

    Thompson, Stuart A; Gaynor, Erin C

    2008-11-13

    Mechanisms underlying virulence properties of Campylobacter jejuni have historically been difficult to identify. In this issue of Cell Host & Microbe, Hofreuter et al. (2008) show that C. jejuni's ability to metabolize glutamine, glutathione, and asparagine affects its ability to colonize specific host tissues. These findings reflect the emerging theme of bacterial physiology directly impacting pathogenesis. PMID:18996338

  18. Campylobacter jejuni Host Tissue Tropism: A Consequence of Its Low-Carb Lifestyle?

    OpenAIRE

    Thompson, Stuart A.; Gaynor, Erin C.

    2008-01-01

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

  19. Nucleases Encoded by Integraded Elements CJIE2 and CJIE4 Inhibit Natural Transformation of Campylobacter Jejuni

    NARCIS (Netherlands)

    Gaasbeek, E.J.; Wagenaar, J.A.; Guilhabert, M.R.; Putten, van J.P.; Parker, C.T.; Wal, van der F.J.

    2010-01-01

    The species Campylobacter jejuni is naturally competent for DNA uptake; nevertheless, nonnaturally transformable strains do exist. For a subset of strains we previously showed that a periplasmic DNase, encoded by dns, inhibits natural transformation in C. jejuni. In the present study, genetic factor

  20. Genome sequences of two stress-tolerant Campylobacter jejuni poultry strains, 305 and DFVF1099

    DEFF Research Database (Denmark)

    Takamiya, Monica; Özen, Asli Ismihan; Rasmussen, Morten;

    2011-01-01

    Campylobacter jejuni is a food-borne pathogen with a high prevalence in poultry meat, which in fresh unfrozen condition is the major source of campylobacteriosis. C. jejuni strains DFVF1099 and 305 are considered tolerant to several environmental stresses (T. Birk et al., J. Food Prot. 73...

  1. Energy Taxis Drives Campylobacter jejuni toward the Most Favorable Conditions for Growth

    DEFF Research Database (Denmark)

    Vegge, C.S.; Brondsted, L.; Li, Yiping;

    2009-01-01

    Campylobacter jejuni is a serious food-borne bacterial pathogen in the developed world. Poultry is a major reservoir, and C. jejuni appears highly adapted to the gastrointestinal tract of birds. Several factors are important for chicken colonization and virulence, including a taxis mechanism...

  2. Composting poultry manure by fly larvae (Musca domestica) eliminates Campylobacter jejuni from the manure

    OpenAIRE

    Nordentoft, Steen; Hald, Birthe

    2013-01-01

    Introduction The common house fly, Musca domestica (Md) is an important carrier of zoonotic agents, and Campylobacter jejuni is one that may be transmitted between animals and humans by flies. Colonized animals shed the bacteria in feces where larval stages of Md flies develops. Aim of the present study To monitor fly larvae composting of poultry manure artificially contaminated with C. jejuni, and to investigate a possible transmission route of C. jejuni from the manure through the fly larva...

  3. Comparison of Campylobacter jejuni Lipooligosaccharide Biosynthesis Loci from a Variety of Sources

    OpenAIRE

    Craig T Parker; Horn, Sharon T.; Gilbert, Michel; Miller, William G.; Woodward, David L.; Mandrell, Robert E.

    2005-01-01

    Campylobacter jejuni strains exhibit significant variation in the genetic content of the lipooligosaccharide (LOS) biosynthesis loci with concomitant differences in LOS structure. The C. jejuni LOS loci have been grouped into six classes based on gene content and organization. Utilizing PCR amplifications of genes from these loci, we were able to classify a majority (80%) of the LOS biosynthesis loci from 123 strains of C. jejuni that included 39 of the Penner serotype reference strains. We f...

  4. Neonatal sepsis by campylobacter jejuni: a genetically proven transmission from a household puppy

    NARCIS (Netherlands)

    Wolfs, T.F.W.; Duim, B.; Geelen, S.P.M.; Rigter, A.; Thomson Carter, F.; Fleer, A.; Wagenaar, J.A.

    2001-01-01

    We report a case of neonatal Campylobacter jejuni sepsis in a 3-week-old infant who acquired the infection through transmission from a recently acquired household puppy. Genotyping of Campylobacter strains obtained from puppy and child resulted in highly homogeneous findings. This represents the fir

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

    Science.gov (United States)

    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...

  6. Neonatal sepsis by Campylobacter jejuni : Genetically proven transmission from a household puppy

    NARCIS (Netherlands)

    Wolfs, TFW; Duim, B; Geelen, SPM; Rigter, A; Thomson-Carter, F; Fleer, A; Wagenaar, JA

    2001-01-01

    We report a case of neonatal Campylobacter jejuni sepsis in a 3-week-old infant who acquired the infection through transmission from a recently acquired household puppy. Genotyping of Campylobacter strains obtained from puppy and child resulted in highly homogenous findings. This represents the firs

  7. Antimicrobial activity of essential oils and five terpenoid compounds against Campylobacter jejuni in pure and mixed culture experiments.

    Science.gov (United States)

    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

  8. Antimicrobial activity of essential oils and five terpenoid compounds against Campylobacter jejuni in pure and mixed culture experiments.

    Science.gov (United States)

    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.

  9. Isolation of Campylobacter from human stool samples

    Directory of Open Access Journals (Sweden)

    S M Salim

    2014-01-01

    Full Text Available Context: Campylobacter is an undetected cause of diarrhoea especially under 5 years of age in most of the countries. Isolation of this organism is difficult, expensive and cumbersome. Aims: Our objective of this study was to isolate this pathogen from the stool specimens on routinely available blood containing laboratory media using the candle jar for creating the microaerophilic atmosphere in our setup. Settings and Designs: A descriptive study. Materials and Methods: A total of 50 stool samples were inoculated onto selective and non-selective media with and without filtration using a 0.45 μm membrane. The inoculated media were simultaneously incubated in microaerophilic conditions using the Anoxomat as well as in candle jars at temperatures 37°C and 42°C. The culture isolates were confirmed by standard phenotypic tests. A simplex polymerase chain reaction (PCR targeting the 16S ribosomal deoxyribonucleic acid of Campylobacter was performed on the deoxyribonucleic acid (DNA of the culture isolates as well as on the DNA extracted from the stool filtrates. Statistical Analysis: Data was expressed as a proportion. Results: Campylobacter could be isolated in 5 out of 50 stool samples using both the Anoxomat as well as the candle jar. Furthermore, we did not find any difference between the isolation using the selective and blood containing media as well as the different incubation temperatures. All the five were confirmed phenotypically and genotypically to be Campylobacter jejuni. The PCR results corroborated with that of the culture. Conclusions: Isolation by culture was as sensitive as that of the PCR.

  10. Phase variable expression of capsular polysaccharide modifications allows Campylobacter jejuni to avoid bacteriophage infection in chickens

    Directory of Open Access Journals (Sweden)

    Martine Camilla Holst Sørensen

    2012-02-01

    Full Text Available Bacteriophages are estimated to be the most abundant entities on earth and can be found in every niche where their bacterial hosts reside. The initial interaction between phages and Campylobacter jejuni, a common coloniser of poultry intestines and a major source of foodborne bacterial gastroenteritis in humans, is not well understood. Recently, we isolated and characterised a phage F336 resistant variant of C. jejuni NCTC11168 called 11168R. Comparisons of 11168R with the wildtype lead to the identification of a novel phage receptor, the phase variable O-methyl phosphoramidate (MeOPN moiety of the C. jejuni capsular polysaccharide (CPS. In this study we demonstrate that the 11168R strain has gained cross-resistance to four other phages in our collection (F198, F287, F303 and F326. The reduced plaquing efficiencies suggested that MeOPN is recognized as a receptor by several phages infecting C. jejuni. To further explore the role of CPS modifications in C. jejuni phage recognition and infectivity, we tested the ability of F198, F287, F303, F326 and F336 to infect different CPS variants of NCTC11168, including defined CPS mutants. These strains were characterised by high-resolution magic angle spinning NMR spectroscopy. We found that in addition to MeOPN, the phase variable 3-O-Me and 6-O-Me groups of the NCTC11168 CPS structure may influence the plaquing efficiencies of the phages. Furthermore, co-infection of chickens with both C. jejuni NCTC11168 and phage F336 resulted in selection of resistant C. jejuni bacteria, which either lack MeOPN or gain 6-O-Me groups on their surface, demonstrating that resistance can be acquired in vivo. In summary, we have shown that phase variable CPS structures modulate phage infectivity in C. jejuni and suggest that the constant phage predation in the avian gut selects for changes in these structures leading to a continuing phage-host co-evolution.

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

    Science.gov (United States)

    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.

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

    Directory of Open Access Journals (Sweden)

    Maike T. Maziero

    2010-06-01

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

  13. Source attribution of human Campylobacter isolates by MLST and fla-typing and association of genotypes with quinolone resistance

    OpenAIRE

    Kittl, Sonja; Heckel, Gerald; Korczak, Bozena; Kuhnert, Peter

    2013-01-01

    Campylobacteriosis is the most frequent zoonosis in developed countries and various domestic animals can function as reservoir for the main pathogens Campylobacter jejuni and Campylobacter coli. In the present study we compared population structures of 730 C. jejuni and C. coli from human cases, 610 chicken, 159 dog, 360 pig and 23 cattle isolates collected between 2001 and 2012 in Switzerland. All isolates had been typed with multi locus sequence typing (MLST) and flaB-typing and their genot...

  14. Source Attribution of Human Campylobacter Isolates by MLST and Fla-Typing and Association of Genotypes with Quinolone Resistance

    OpenAIRE

    Sonja Kittl; Gerald Heckel; Korczak, Bożena M.; Peter Kuhnert

    2013-01-01

    Campylobacteriosis is the most frequent zoonosis in developed countries and various domestic animals can function as reservoir for the main pathogens Campylobacter jejuni and Campylobacter coli. In the present study we compared population structures of 730 C. jejuni and C. coli from human cases, 610 chicken, 159 dog, 360 pig and 23 cattle isolates collected between 2001 and 2012 in Switzerland. All isolates had been typed with multi locus sequence typing (MLST) and flaB-typing and their genot...

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

    Science.gov (United States)

    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

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

    OpenAIRE

    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.

  17. Campylobacter jejuni occurrence in chicken fecal samples from small properties in Pelotas, southern of Brazil Ocorrência de Campylobacter jejuni em amostras fecais de galinhas em pequenas propriedades de Pelotas, sul do Brasil

    Directory of Open Access Journals (Sweden)

    Fabiane R. Gomes

    2006-09-01

    Full Text Available The aim of this study was to evaluate the occurrence of thermophilic Campylobacter species in broiler flocks from 26 small properties located near the campus of Federal University of Pelotas, RS, Brazil. A total of 404 chicken fecal samples were analyzed and after isolation, identification and biotyping was performed according to Lior´s scheme. Twenty one strains (5.2% of Campylobacter jejuni biotype II were isolated from animals from seven properties (26.9%, with a variation from 3% to 34.6% among the positive properties. We also analyzed twelve C. jejuni isolates by polymerase chain reaction (PCR, using specific primers that generate a fragment of 402 pb. Our study allowed us to observe that chickens raised in non-industrial establishments without sanitary attention can harbour C. jejuni in their intestinal tract, and by doing so, they could be a serious health risk to humans.O objetivo deste estudo foi avaliar a ocorrência de espécies termofílicas de Campylobacter em galinhas de 26 pequenas propriedades localizadas próximo ao campus da Universidade Federal de Pelotas, RS, Brasil. Um total de 404 amostras fecais de galinhas foram analisadas e após o isolamento, a identificação e a biotipificação foram realizadas de acordo com o esquema de Lior. Vinte e uma amostras (5,2% de Campylobacter jejuni biotipo II foram isoladas de animais de sete propriedades (26,9%, com uma variação de 3,3% a 34,6% entre as propriedades positivas. Primers específicos, que geram um fragmento de 402pb, foram usados em PCR para analisar 12 isolados de C. jejuni. Concluímos que galinhas criadas em estabelecimentos não industriais e sem atenção sanitária, são portadoras de C. jejuni e, desta forma, podem ser consideradas um fator de risco para infecção humana.

  18. Host-Pathogen Interactions in Guillain-Barré Syndrome : the role of Campylobacter jejuni lipooligosaccharide sialylation

    NARCIS (Netherlands)

    A.P. Heikema (Astrid)

    2013-01-01

    markdownabstract*Campylobacter jejuni* (*C. jejuni*) is a spiral, comma-shaped Gram-negative bacterium which is motile due to bipolar flagella. *C. jejuni* is frequently present in the intestines of poultry and birds, where it is considered to be part of the normal intestinal flora (1). During slaug

  19. Intranasal Immunization with Chitosan/pCAGGS-flaA Nanoparticles Inhibits Campylobacter jejuni in a White Leghorn Model

    Directory of Open Access Journals (Sweden)

    Jin-lin Huang

    2010-01-01

    Full Text Available Campylobacter jejuni is the most common zoonotic bacterium associated with human diarrhea, and chickens are considered to be one of the most important sources for human infection, with no effective prophylactic treatment available. We describe here a prophylactic strategy using chitosan-DNA intranasal immunization to induce specific immune responses. The chitosan used for intranasal administration is a natural mucus absorption enhancer, which results in transgenic DNA expression in chicken nasopharynx. Chickens immunized with chitosan-DNA nanoparticles, which carried a gene for the major structural protein FlaA, produced significantly increased levels of serum anti-Campylobacter jejuni IgG and intestinal mucosal antibody (IgA, compared to those treated with chitosan-DNA (pCAGGS. Chitosan-pCAGGS-flaA intranasal immunization induced reductions of bacterial expellation by 2-3 log10 and 2 log10 in large intestine and cecum of chickens, respectively, when administered with the isolated C. jejuni strain. This study demonstrated that intranasal delivery of chitosan-DNA vaccine successfully induced effective immune response and might be a promising vaccine candidate against C. jejuni infection.

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

    OpenAIRE

    Olofsson, Jenny; Griekspoor Berglund, Petra; 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. co...

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

    Science.gov (United States)

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

    2015-01-01

    The present study was conducted to investigate the prevalence and antibiotic resistance among Campylobacter jejuni in ulam at farms and retail outlets located in Kuala Terengganu, Malaysia. A total of 526 samples (ulam, soil, and fertilizer) were investigated for the presence of C. jejuni and the gene for cytolethal distending toxin (cdt) by using a multiplex PCR method. Antibiotic susceptibility to 10 types of antibiotics was determined using the disk diffusion method for 33 C. jejuni isolates. The average prevalence of contaminated samples from farms, wet markets, and supermarkets was 35.29, 52.66, and 69.88%, respectively. The cdt gene was not detected in 24 of the 33 C. jejuni isolates, but 9 isolates harbored cdtC. Antibiotic resistance in C. jejuni isolates was highest to penicillin G (96.97% of isolates) followed by vancomycin (87.88%), ampicillin (75.76%), erythromycin (60.61%), tetracycline (9.09%), amikacin (6.06%), and norfloxacin (3.03%); none of the isolates were resistant to ciprofloxacin, enrofloxacin, and gentamicin. In this study, C. jejuni was present in ulam, and some isolates were highly resistant to some antibiotics but not to quinolones. Thus, appropriate attention and measures are required to prevent C. jejuni contamination on farms and at retail outlets. PMID:25581179

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

    OpenAIRE

    Guillou, Sandrine; Leguérinel, Ivan; Garrec, N.; Renard, M. A.; Cappelier, Jean-Michel; Federighi, Michel

    2008-01-01

    International audience 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 1C i...

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

    Science.gov (United States)

    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.

  4. Genome and Proteome of Campylobacter jejuni Bacteriophage NCTC 12673▿†

    OpenAIRE

    Kropinski, Andrew M.; Arutyunov, Denis; Foss, Mary; Cunningham, Anna; Ding, Wen; Singh, Amit; Pavlov, Andrey R.; Henry, Matthew; Evoy, Stephane; Kelly, John; Szymanski, Christine M.

    2011-01-01

    Campylobacter jejuni continues to be the leading cause of bacterial food-borne illness worldwide, so improvements to current methods used for bacterial detection and disease prevention are needed. We describe here the genome and proteome of C. jejuni bacteriophage NCTC 12673 and the exploitation of its receptor-binding protein for specific bacterial detection. Remarkably, the 135-kb Myoviridae genome of NCTC 12673 differs greatly from any other proteobacterial phage genome described (includin...

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

    Science.gov (United States)

    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

  6. Enhanced biofilm formation and multi-host transmission evolve from divergent genetic backgrounds in Campylobacter jejuni.

    Science.gov (United States)

    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.

  7. Study of the infectivity of saline-stored Campylobacter jejuni for day-old chicks

    DEFF Research Database (Denmark)

    Hald, Birthe; Knudsen, Katrine; Lind, Peter;

    2001-01-01

    The culturability of three Campylobacter jejuni strains and their infectivity for day-old chicks were assessed following storage of the strains in saline. The potential for colonization of chicks was weakened during the storage period and terminated 3 to 1 weeks before the strains became noncultu......The culturability of three Campylobacter jejuni strains and their infectivity for day-old chicks were assessed following storage of the strains in saline. The potential for colonization of chicks was weakened during the storage period and terminated 3 to 1 weeks before the strains became......-campylobacter outer membrane protein serum antibodies in day-old chicks did not protect the chicks from campylobacter colonization....

  8. Genomotype Analyses for the Investigation of Campylobacter spp. Isolates with Distinct flaA Alleles of Recovered from Iceland

    Science.gov (United States)

    The Gram-negative bacteria, Campylobacter jejuni, is the leading bacterial etiology of acute gastroenteritis in humans. Evidence implicates poultry as a potential source of the organism for human illness. Campylobacter spp. isolates vary in their virulence properties and recent comparative phyloge...

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

    OpenAIRE

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

    2011-01-01

    Campylobacter jejuni is a prevalent gastrointestinal pathogen in humans and a common commensal of poultry. When colonizing its hosts, C. jejuni comes into contact with intestinal carbohydrates, including l-fucose, released from mucin glycoproteins. Several strains of C. jejuni possess a genomic island (cj0480c–cj0490) that is up-regulated in the presence of both l-fucose and mucin and allows for the utilization of l-fucose as a substrate for growth. Strains possessing this genomic island show...

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

    DEFF Research Database (Denmark)

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

    2006-01-01

    The effect of a Campylobacter jejuni colonization on the development of the microflora of the cecum and the ileum of broiler chickens was studied using molecular methods. The infection did affect the development and complexity of the microbial Communities of the ceca, but we found no permanent...... effect of a C. jejuni infection on the ileal microflora of the broilers. In addition, denaturant gradient gel electrophoresis (DGGE) profiles generated from cecal and ileal contents revealed several DGGE bands that were present in the control chickens, but not in the chickens colonized with C. jejuni...

  11. Host epithelial cell invasion by Campylobacter jejuni: trigger or zipper mechanism?

    Directory of Open Access Journals (Sweden)

    Tadhg eÓ Cróinín

    2012-03-01

    Full Text Available Campylobacter jejuni, a spiral-shaped Gram-negative pathogen, is a highly frequent cause of gastrointestinal foodborne illness in humans worldwide. Clinical outcome of C. jejuni infections ranges from mild to severe diarrheal disease, and some other complications including reactive arthritis and Guillain–Barré syndrome. This review article highlights various C. jejuni pathogenicity factors, host cell determinants and proposed signaling mechanisms involved in human host cell invasion and their potential role in the development of C. jejuni-mediated disease. A model is presented which outlines the various important interactions of C. jejuni with the intestinal epithelium, and we discuss the pro’s and con’s for the zipper over the trigger mechanism of invasion. Future work should clarify the contradictory role of some previously identified factors, and should identify and characterize novel virulence determinants, which are crucial to provide fresh insights into the diversity of strategies employed by this pathogen to cause disease.

  12. Defense and adaptation: The complex inter-relationship between Campylobacter jejuni and mucus.

    Directory of Open Access Journals (Sweden)

    Abofu eAlemka

    2012-02-01

    Full Text Available Mucus colonization is the first step towards the establishment of infection and disease by mucosal pathogens. There is an emerging literature implicating specific mucin subtypes and mucin modifications in protecting the host from Campylobacter jejuni infection. However, mucosal pathogens have evolved sophisticated mechanisms to breach the mucus layer and C. jejuni in particular appears to harbor specific adaptations to better colonize intestinal mucus. For example, components of mucus are chemotactic for C. jejuni and the rheological properties of mucus promote motility of the organism. Furthermore, recent studies demonstrate that mucins modulate the pathogenicity of C. jejuni in a species-specific manner and likely help determine whether these bacteria become pathogenic (as in humans, or adopt a commensal mode of existence (as in chickens and other animals. This review focuses on recent advances in understanding the complex interplay between C. jejuni and components of the mucus layer.

  13. Humoral immune response to campylobacter jejuni in patients with enterocolitis and Guillain-Barré syndrome

    Directory of Open Access Journals (Sweden)

    Ristić Ljiljana

    2012-01-01

    Full Text Available Campylobacter jejuni is one of the most important causes of diarrheal disease worldwide. In addition, it can cause neurological post-infectious sequels, such as Guillain-Barré syndrome (GBS. Humoral immune response to C. jejuni was monitored in patients with C. jejuni enterocolitis, GBS patients and healthy persons, by ELISA. Statistical significance between patients with enterocolitis and healthy persons, as well as among GBS patients and healthy controls, was proven. Statistical significance in IgA among the examined groups was also noticed. The highest values of IgM were found in the patients with GBS, while the highest values of IgG were found in those with enterocolitis. C. jejuni is a significant cause of antecedent infection in GBS. ELISA techniques can be considered a reliable method in determining the presence of serum antibodies in patients with enterocolitis caused by C. jejuni, as well as in patients with GBS.

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

    Science.gov (United States)

    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.

  15. Campylobacter

    NARCIS (Netherlands)

    Wagenaar, J.A.

    2015-01-01

    Campylobacteriosis is a frequently diagnosed disease in humans. Most infections are considered food-borne and are caused by Campylobacter jejuni and C. coli. The animal reservoirs of these Campylobacter, and the sources and routes of transmission, are described and discussed. Most warm-blooded an

  16. Outer membrane vesicle-mediated release of cytolethal distending toxin (CDT from Campylobacter jejuni

    Directory of Open Access Journals (Sweden)

    Uhlin Bernt

    2009-10-01

    Full Text Available Abstract Background Background: Cytolethal distending toxin (CDT is one of the well-characterized virulence factors of Campylobacter jejuni, but it is unknown how CDT becomes surface-exposed or is released from the bacterium to the surrounding environment. Results Our data suggest that CDT is secreted to the bacterial culture supernatant via outer membrane vesicles (OMVs released from the bacteria. All three subunits (the CdtA, CdtB, and CdtC proteins were detected by immunogold labeling and electron microscopy of OMVs. Subcellular fractionation of the bacteria indicated that, apart from the majority of CDT detected in the cytoplasmic compartment, appreciable amounts (20-50% of the cellular pool of CDT proteins were present in the periplasmic compartment. In the bacterial culture supernatant, we found that a majority of the extracellular CDT was tightly associated with the OMVs. Isolated OMVs could exert the cell distending effects typical of CDT on a human intestinal cell line, indicating that CDT is present there in a biologically active form. Conclusion Our results strongly suggest that the release of outer membrane vesicles is functioning as a route of C. jejuni to deliver all the subunits of CDT toxin (CdtA, CdtB, and CdtC to the surrounding environment, including infected host tissue.

  17. Campylobacter jejuni induces transcellular translocation of commensal bacteria via lipid rafts

    Directory of Open Access Journals (Sweden)

    Kalischuk Lisa D

    2009-02-01

    Full Text Available Abstract Background Campylobacter enteritis represents a risk factor for the development of inflammatory bowel disease (IBD via unknown mechanisms. As IBD patients exhibit inflammatory responses to their commensal intestinal microflora, factors that induce translocation of commensal bacteria across the intestinal epithelium may contribute to IBD pathogenesis. This study sought to determine whether Campylobacter induces translocation of non-invasive intestinal bacteria, and characterize underlying mechanisms. Methods Mice were infected with C. jejuni and translocation of intestinal bacteria was assessed by quantitative bacterial culture of mesenteric lymph nodes (MLNs, liver, and spleen. To examine mechanisms of Campylobacter-induced bacterial translocation, transwell-grown T84 monolayers were inoculated with non-invasive Escherichia coli HB101 ± wild-type Campylobacter or invasion-defective mutants, and bacterial internalization and translocation were measured. Epithelial permeability was assessed by measuring flux of a 3 kDa dextran probe. The role of lipid rafts was assessed by cholesterol depletion and caveolin co-localization. Results C. jejuni 81–176 induced translocation of commensal intestinal bacteria to the MLNs, liver, and spleen of infected mice. In T84 monolayers, Campylobacter-induced internalization and translocation of E. coli occurred via a transcellular pathway, without increasing epithelial permeability, and was blocked by depletion of epithelial plasma membrane cholesterol. Invasion-defective mutants and Campylobacter-conditioned cell culture medium also induced E. coli translocation, indicating that C. jejuni does not directly 'shuttle' bacteria into enterocytes. In C. jejuni-treated monolayers, translocating E. coli associated with lipid rafts, and this phenomenon was blocked by cholesterol depletion. Conclusion Campylobacter, regardless of its own invasiveness, promotes the translocation of non-invasive bacteria across

  18. Antimicrobial edible apple films inactivate antibiotic resistant and susceptible Campylobacter jejuni strains on chicken breast

    Science.gov (United States)

    Campylobacter jejuni is the leading cause of bacterial diarrheal illness worldwide. Many strains are now becoming multi-drug resistant. To help overcome this problem, apple-based edible films containing carvacrol and cinnamaldehyde were evaluated for their effectiveness against antibiotic resistant...

  19. Intracellular pH Campylobacter jejuni when treated with aqueous chlorine dioxide

    DEFF Research Database (Denmark)

    Smigic, Nada; Rajkovic, Andreja; Arneborg, Nils;

    2011-01-01

    The aim of this study was to investigate the response of Campylobacter jejuni at single-cell level when exposed to different concentrations of chlorine dioxide (ClO2). The parameter of choice, intracellular pH (pHi), was determined by using fluorescence ratio imaging microscopy with a pH-sensitiv...

  20. Serotype and genotype diversity and hatchery transmission of Campylobacter jejuni in commercial poultry flocks

    DEFF Research Database (Denmark)

    Petersen, L.; Nielsen, E.M.; On, Stephen L.W.

    2001-01-01

    or horizontal transmission via the hatchery are not significant transmission routes of C jejuni to broiler chickens under Danish conditions. In the cases where more than one Campylobacter clone simultaneously colonised flocks, we found that the different clones coexisted in flocks rather than excluding each...

  1. Duration of the vector period of house flies for Campylobacter jejuni estimated by experimental infection series

    DEFF Research Database (Denmark)

    Skovgård, Henrik; Hald, Birthe

    The house fly Musca domestica may act as a vector and transmit Campylobacter to broiler flocks. We determined the duration of the vector period for C. jejuni at various temperatures and inoculation doses. For the temperature experiment, laboratory reared house flies (n = 375) were inoculated with...

  2. Composting poultry manure by fly larvae (Musca domestica) eliminates Campylobacter jejuni from the manure

    DEFF Research Database (Denmark)

    Nordentoft, Steen; Hald, Birthe

    2013-01-01

    Introduction The common house fly, Musca domestica (Md) is an important carrier of zoonotic agents, and Campylobacter jejuni is one that may be transmitted between animals and humans by flies. Colonized animals shed the bacteria in feces where larval stages of Md flies develops. Aim of the present...

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

    Science.gov (United States)

    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...

  4. Colonisation of a phage susceptible Campylobacter jejuni population in two phage positive broiler flocks.

    Directory of Open Access Journals (Sweden)

    Sophie Kittler

    Full Text Available The pathogens Campylobacter jejuni and Campylobacter coli are commensals in the poultry intestine and campylobacteriosis is one of the most frequent foodborne diseases in developed and developing countries. Phages were identified to be effective in reducing intestinal Campylobacter load and this was evaluated, in the first field trials which were recently carried out. The aim of this study was to further investigate Campylobacter population dynamics during phage application on a commercial broiler farm. This study determines the superiority in colonisation of a Campylobacter type found in a field trial that was susceptible to phages in in vitro tests. The colonisation factors, i.e. motility and gamma glutamyl transferase activity, were increased in this type. The clustering in phylogenetic comparisons of MALDI-TOF spectra did not match the ST, biochemical phenotype and phage susceptibility. Occurrence of Campylobacter jejuni strains and phage susceptibility types with different colonisation potential seem to play a very important role in the success of phage therapy in commercial broiler houses. Thus, mechanisms of both, phage susceptibility and Campylobacter colonisation should be further investigated and considered when composing phage cocktails.

  5. Cytokine responses in primary chicken embryo intestinal cells infected with Campylobacter jejuni strains of human and chicken origin and the expression of bacterial virulence-associated genes

    DEFF Research Database (Denmark)

    Li, Yiping; Ingmer, Hanne; Madsen, Mogens;

    2008-01-01

    Background Campylobacter jejuni is a major cause of inflammatory diarrhoea in humans and is considered a commensal of the gastroenteric tract of the avian host. However, little is known about the interaction between C. jejuni and the avian host including the cytokine responses and the expression....... jejuni strains are capable of invading the CEICs and stimulate these cells in a pro-inflammatory manner and during this interaction the expression of the bacterial virulence-associated genes ciaB, dnaJ and racR is increased. Furthermore, incubation of bacteria with conditioned cell- and bacteria......-free media from another co-cultivation experiment also increased the expression of the virulence-associated genes in the C. jejuni chicken isolate, indicating that the expression of bacterial genes is regulated by component(s) secreted upon co-cultivation of bacteria and CEICs. Conclusion We show that under...

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

    Science.gov (United States)

    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

  7. Proteomic and genomic analysis reveals novel Campylobacter jejuni outer membrane proteins and potential heterogeneity

    Directory of Open Access Journals (Sweden)

    Eleanor Watson

    2014-09-01

    Full Text Available Gram-negative bacterial outer membrane proteins play important roles in the interaction of bacteria with their environment including nutrient acquisition, adhesion and invasion, and antibiotic resistance. In this study we identified 47 proteins within the Sarkosyl-insoluble fraction of Campylobacter jejuni 81-176, using LC–ESI-MS/MS. Comparative analysis of outer membrane protein sequences was visualised to reveal protein distribution within a panel of Campylobacter spp., identifying several C. jejuni-specific proteins. Smith–Waterman analyses of C. jejuni homologues revealed high sequence conservation amongst a number of hypothetical proteins, sequence heterogeneity of other proteins and several proteins which are absent in a proportion of strains.

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

    Directory of Open Access Journals (Sweden)

    Soo Hyeon Hong

    2016-06-01

    Full Text Available 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.

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

    Science.gov (United States)

    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 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

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

    Science.gov (United States)

    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

  11. Assess the prevalence rate of Campylobacter genus and Campylobacter jejuni species in raw milk collected from the Amol City by Multiplex- Polymerase Chain Reaction

    Directory of Open Access Journals (Sweden)

    Ali Dabiri

    2016-03-01

    Full Text Available Background & Objective: Campylobacter can be transmitted through the raw milk. The purpose of this study was to determine the prevalence of Campylobacter genus and Campylobacter jejuni (C. jejuni species in raw milk samples. Materials & Methods: In this study, 72 samples of raw milk were collected of the platforms milk in the Amol city in summer. Phenotypic identification of Campylobacter genus and C. jejuni species using microbiology laboratory methods and molecular identification of this bacterium using Multiplex- Polymerase Chain Reaction (M-PCR were performed. The data was calculated using the SPSS 16.0 software and the Fisher's exact test (p < 0.05. Results: Among the 72 samples, 13.88% of samples were contaminated with C. jejuni and 2.77% were contaminated with Campylobacter genus. The highest prevalence rate for this bacterium was in July (20.83% and the lowest prevalence rate was in September (12.5%. The significant difference between the prevalence of the Campylobacter genus and C. jejuni species in raw milk samples in various months of summer was not observed (p = 0.07. Conclusion: This study showed the raw milk contamination with Campylobacter, and thereby the sanitation in the dairy food production places and the use of fast and accurate method to identify this bacterium is important.

  12. Disruption of colonic barrier function and induction of mediator release by strains of Campylobacter jejuni that invade epithelial cells

    Institute of Scientific and Technical Information of China (English)

    Johannes Beltinger; Jo del Buono; Maeve M Skelly; John Thornley; Robin C Spiller; William A Stack; Christopher J Hawkey

    2008-01-01

    AIM:To study the mechanisms by which Campylobacter jejuni (C.jejuni) causes inflammation and diarrhea.In particular,direct interactions with intestinal epithelial cells and effects on barrier function are poorly understood.METHODS:To model the initial pathogenic effects of C.jejuni on intestinal epithelium,polarized human colonic HCA-7 monolayerswere grown on permeabilized filters and infected apically with clinical isolates of C.jejuni.Integrity of the monolayer was monitored by changes in monolayer resistance,release of lactate dehydrogenase,mannitol fluxes and electron microscopy.Invasion of HCA-7 cells was assessed by a modified gentamicin protection assay,translocation by counting colony forming units in the basal chamber,stimulation of mediator release by immunoassays and secretory responses in monolayers stimulated by bradykinin in an Ussing chamber.RESULTS:All strains translocated across monolayers but only a minority invaded HCA-7 cells.Strains that invaded HCA-7 cells destroyed rnonolayer resistance over 6 h,accompanied by increased release of lactate dehydrogenase,a four-fold increase in permeability to [3H] mannitol,and ultrastructural disruption of tight junctions,with rounding and lifting of cells off the filter membrane.Synthesis of interleukin (IL)-8 and prostaglandin E2 was increased with strains that invaded the rnonolayer but not with those that did not.CONCLUSION:These data demonstrate two distinct effects of C.jejuni on colonic epithelial cells and provide an informative model for further investigation of initial host cell responses to C.jejuni.

  13. Guillain-Barré syndrome- and Miller Fisher syndrome-associated Campylobacter jejuni lipopolysaccharides induce anti-GM1 and anti-GQ1b Antibodies in rabbits.

    NARCIS (Netherlands)

    M.A. de Klerk; H.P. Endtz (Hubert); B.C. Jacobs (Bart); J.D. Laman (Jon); F.G.A. van der Meché; P.A. van Doorn (Pieter); C.W. Ang (Wim)

    2001-01-01

    textabstractCampylobacter jejuni infections are thought to induce antiganglioside antibodies in patients with Guillain-Barre syndrome (GBS) and Miller Fisher syndrome (MFS) by molecular mimicry between C. jejuni lipopolysaccharides (LPS) and gangliosides. We used purifi

  14. Natural transformation of Campylobacter jejuni occurs beyond limits of growth

    DEFF Research Database (Denmark)

    Vegge, Christina Skovgaard; Brøndsted, Lone; Ligowska, Małgorzata;

    2012-01-01

    to be one of the main mechanisms for mediating transfer of genetic material and evolution of the organism. Given the diverse habitats of C. jejuni we set out to examine how environmental conditions and physiological processes affect natural transformation of C. jejuni. We show that the efficiency...... of transformation is correlated to the growth conditions, but more importantly that transformation occurs at growth-restrictive conditions as well as in the late stationary phase; hence revealing that growth per se is not required for C. jejuni to be competent. Yet, natural transformation of C. jejuni is an energy...

  15. cj0371: a novel virulence-associated gene of Campylobacter jejuni

    Directory of Open Access Journals (Sweden)

    Xueqing Du

    2016-07-01

    Full Text Available 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.

  16. Survival of Campylobacter jejuni under Conditions of Atmospheric Oxygen Tension with the Support of Pseudomonas spp.▿

    OpenAIRE

    Hilbert, Friederike; Scherwitzel, Manuela; Paulsen, Peter; Szostak, Michael P

    2010-01-01

    Campylobacter jejuni is a major food-borne pathogen. Despite causing enteritis in humans, it is a well-adapted intestinal microorganism in animals, hardly ever generating disease symptoms. Nevertheless, as a true microaerophilic microorganism it is still puzzling how Campylobacter cells can survive on chicken meat, the main source of human infection. In this study, we demonstrate that C. jejuni is able to withstand conditions of atmospheric oxygen tension when cocultured with Pseudomonas spec...

  17. Prevalence, antibiotic resistance and RAPD typing of Campylobacter species isolated from ducks, their rearing and processing environments in Penang, Malaysia.

    Science.gov (United States)

    Adzitey, Frederick; Rusul, Gulam; Huda, Nurul; Cogan, Tristan; Corry, Janet

    2012-03-15

    We report for the first time on the prevalence, antibiotic resistance and RAPD types of Campylobacter species in ducks and duck related environmental samples in Malaysia. Samples were examined by enrichment in Bolton Broth followed by plating onto modified Charcoal Cefoperazone Deoxycholate agar (mCCDA) and/or plating directly onto mCCDA. A total of 643 samples were screened, and the prevalence of Campylobacter spp. in samples from different sources ranged from 0% to 85%. The method of isolation had a significant (P<0.05) effect on the isolation rate. One hundred and sixteen Campylobacter isolates, comprising of 94 Campylobacter jejuni, 19 Campylobacter coli and three Campylobacter lari, were examined for their sensitivity to 13 antibiotics. Majority of the C. jejuni isolates were resistant to cephalothin (99%), tetracycline (96%), suphamethoxazole/trimethoprim (96%), and very few were resistant to gentamicin (5%), chloramphenicol (7%) and erythromycin (1%). All C. coli isolates were resistant to cephalothin, nalidixic acid, norfloxacin and tetracycline but susceptible to chloramphenicol, erythromycin and gentamicin. The three C. lari isolates were resistant to all the antibiotics tested except chloramphenicol and gentamicin (1/3 and 2/3 susceptible, respectively). Genetic diversity of Campylobacter isolates were determined using random amplification of polymorphic DNA (RAPD). C. jejuni and C. coli isolates belong to fifty-eight and twelve RAPD types, respectively.

  18. The role of Campylobacter jejuni cytolethal distending toxin in gastroenteritis: toxin detection, antibody production, and clinical outcome

    Energy Technology Data Exchange (ETDEWEB)

    Boisen, Nadia [University of Virginia School of Medicine

    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.

  19. Lack of association between the presence of the pVir plasmid and bloody diarrhea in Campylobacter jejuni enteritis.

    NARCIS (Netherlands)

    R.P.L. Louwen (Rogier); A.F. van Belkum (Alex); J.A. Wagenaar (Jaap); Y. Doorduyn; R. Achterberg; H.P. Endtz (Hubert)

    2006-01-01

    textabstractThe main mechanisms by which Campylobacter jejuni causes diarrhea are unknown. In contrast to a recent communication, we report here the absence of an association with the plasmid pVir in patients infected with C. jejuni who developed bloody diarrhea in The Netherlands, and we suggest a

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

    Science.gov (United States)

    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

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

    Science.gov (United States)

    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).

  2. Cytokine responses in primary chicken embryo intestinal cells infected with Campylobacter jejuni strains of human and chicken origin and the expression of bacterial virulence-associated genes

    Directory of Open Access Journals (Sweden)

    Bang Dang D

    2008-06-01

    Full Text Available Abstract Background Campylobacter jejuni is a major cause of inflammatory diarrhoea in humans and is considered a commensal of the gastroenteric tract of the avian host. However, little is known about the interaction between C. jejuni and the avian host including the cytokine responses and the expression of the bacterial genes. We have investigated the invasiveness of primary chicken embryo intestinal cells (CEICs by C. jejuni strains of human and chicken origins and the production of pro-inflammatory cytokines as well as the expression of the bacterial virulence-associated genes during co-cultivation. Results C. jejuni strains are capable of invading the CEICs and stimulate these cells in a pro-inflammatory manner and during this interaction the expression of the bacterial virulence-associated genes ciaB, dnaJ and racR is increased. Furthermore, incubation of bacteria with conditioned cell- and bacteria-free media from another co-cultivation experiment also increased the expression of the virulence-associated genes in the C. jejuni chicken isolate, indicating that the expression of bacterial genes is regulated by component(s secreted upon co-cultivation of bacteria and CEICs. Conclusion We show that under in vitro culture condition C. jejuni strains of both human and chicken origins can invade avian host cells with a pro-inflammatory response and that the virulence-associated genes of C. jejuni may play a role in this process.

  3. Relapsing Campylobacter jejuni Systemic Infections in a Child with X-Linked Agammaglobulinemia

    Directory of Open Access Journals (Sweden)

    Paola Ariganello

    2013-01-01

    Full Text Available X-linked agammaglobulinemia (XLA is a primary immunodeficiency of the humoral compartment, due to a mutation in the Bruton tyrosine kinase (BTK gene, characterized by a severe defect of circulating B cells and serum immunoglobulins. Recurrent infections are the main clinical manifestations; although they are especially due to encapsulated bacteria, a specific association with Campylobacter species has been reported. Here, we report the case of a boy with XLA who presented with relapsing Campylobacter jejuni systemic infections. His clinical history supports the hypothesis of the persistence of C. jejuni in his intestinal tract. Indeed, as previously reported, XLA patients may become chronic intestinal carriers of Campylobacter, even in absence of symptoms, with an increased risk of relapsing bacteraemia. The humoral defect is considered to be crucial for this phenomenon, as well as the difficulties to eradicate the pathogen with an appropriate antibiotic therapy; drug resistance is raising in Campylobacter species, and the appropriate duration of treatment has not been established. C. jejuni should always be suspected in XLA patients with signs and symptoms of systemic infection, and treatment should be based on antibiogram to assure the eradication of the pathogen.

  4. Variation of chemosensory receptor content of Campylobacter jejuni strains and modulation of receptor gene expression under different in vivo and in vitro growth conditions

    OpenAIRE

    Day Christopher J; Hartley-Tassell Lauren E; Shewell Lucy K; King Rebecca M; Tram Greg; Day Serena K; Semchenko Evgeny A; Korolik Victoria

    2012-01-01

    Abstract Background Chemotaxis is crucial for the colonisation/infection of hosts with Campylobacter jejuni. Central to chemotaxis are the group A chemotaxis genes that are responsible for sensing the external environment. The distribution of group A chemoreceptor genes, as found in the C. jejuni sequenced strains, tlp1-4, 7, 10 and 11 were determined in 33 clinical human and avian isolates. Results Group A tlp gene content varied among the strains with genes encoding tlp1 (aspartate receptor...

  5. Isolation of Campylobacter from Brazilian broiler flocks using different culturing procedures.

    Science.gov (United States)

    Vaz, C S L; Voss-Rech, D; Pozza, J S; Coldebella, A; Silva, V S

    2014-11-01

    Conventional culturing methods enable the detection of Campylobacter in broiler flocks. However, laboratory culture of Campylobacter is laborious because of its fastidious behavior and the presence of competing nontarget bacteria. This study evaluated different protocols to isolate Campylobacter from broiler litter, feces, and cloacal and drag swabs. Samples taken from commercial Brazilian broiler flocks were directly streaked onto Preston agar (PA), Campy-Line agar (CLA), and modified charcoal cefoperazone deoxycholate agar (mCCDA) and also enriched in blood-free Bolton broth (bfBB) for 24 and 48 h followed by plating onto the different selective media. Higher numbers of Campylobacter-positive cloacal and drag swab samples were observed using either direct plating or enrichment for 24 h before plating onto PA, compared with enrichment for 48 h (P Campylobacter in broiler litter and feces samples. Analysis of directly plated samples revealed that higher Campylobacter levels were detected in feces streaked onto PA (88.8%), cloacal swabs plated onto mCCDA (72.2%), drag swabs streaked onto CLA or mCCDA (69.4%), and litter samples inoculated onto PA (63.8%). Preston agar was the best agar to isolate Campylobacter from directly plated litter samples (P Campylobacter in other samples. The isolated Campylobacter strains were phenotypically identified as Campylobacter jejuni or Campylobacter coli. The predominant contaminant observed in the Campylobacter cultures was Proteus mirabilis, which was resistant to the majority of antimicrobial agents in selective media. Together, these data showed that direct plating onto PA and onto either CLA or mCCDA as the second selective agar enabled the reliable isolation of thermophilic Campylobacter species from broiler samples. Finally, Campylobacter was detected in all broiler flocks sampled.

  6. Crystal structure of JlpA, a surface-exposed lipoprotein adhesin of Campylobacter jejuni

    OpenAIRE

    Kawai, Fumihiro; Paek, Seonghee; Choi, Kyoung-Jae; Prouty, Michael; Kanipes, Margaret I.; Guerry, Patricia; Yeo, Hye-Jeong

    2012-01-01

    The Campylobacter jejuni JlpA protein is a surface-exposed lipoprotein that was discovered as an adhesin promoting interaction with host epithelium cells, an early critical step in the pathogenesis of C. jejuni disease. Increasing evidence ascertained that JlpA is antigenic, indicating a role of JlpA in immune response during the infectious process. Here, we report the crystal structure of JlpA at 2.7Å resolution, revealing a catcher's mitt shaped unclosed half β-barrel. Although the apparent...

  7. A case of myopericarditis associated to Campylobacter jejuni infection in the southern hemisphere

    Directory of Open Access Journals (Sweden)

    Alberto Fica

    2012-06-01

    Full Text Available Myopericarditis is an infrequent complication of acute diarrheal illness due to Campylobacter jejuni, and it has been mainly reported in developed nations. The first case detected in Chile - an upper-middle income country -, that is coincidental with the increasing importance of acute gastroenteritis associated to this pathogen, is described. Recognition of this agent in stools requires special laboratory techniques not widely available, and it was suspected when a young patient presented with acute diarrhea, fever, and chest pain combined with electrocardiogram (EKG abnormalities and elevated myocardial enzymes. C. jejuni myopericarditis can easily be suspected but its detection requires dedicated laboratory techniques.

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

    Directory of Open Access Journals (Sweden)

    Shaun Smith

    2016-06-01

    Full Text Available 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

  9. Reverse transcriptase real-time PCR for detection and quantification of viable Campylobacter jejuni directly from poultry faecal samples

    DEFF Research Database (Denmark)

    Bui, Thanh Xuan; Wolff, Anders; Madsen, Mogens;

    2012-01-01

    and quantification of viable Campylobacter jejuni directly from chicken faecal samples. The results of this method anda DNA-based quantitative real-time PCR (qPCR) method were compared with those of a bacterial culture method. Using bacterial culture andRT-qPCR methods, viable C. jejuni cells could be detected...... detected, and all tested samples were positive, even after 20days of storage. The developed method for detection and quantification of viable C. jejuni cells directly from chicken faecal samples can be usedfor further research on the survival of Campylobacter in the environment....

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

    Directory of Open Access Journals (Sweden)

    Helen L Brown

    2015-07-01

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

  11. Tracing Back Clinical Campylobacter jejuni in the Northwest of Italy and Assessing Their Potential Source.

    Science.gov (United States)

    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

  12. Tracing Back Clinical Campylobacter jejuni in the Northwest of Italy and Assessing Their Potential Source

    Science.gov (United States)

    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

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

    Science.gov (United States)

    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

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

    Science.gov (United States)

    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

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

    Directory of Open Access Journals (Sweden)

    A.V. Karlyshev

    2014-01-01

    Full Text Available 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.

  16. Quantifying Transmission of Campylobacter jejuni in Commercial Broiler Flocks

    NARCIS (Netherlands)

    Gerwe, van T.; Miflin, J.K.; Templeton, J.M.; Bouma, A.; Wagenaar, J.A.; Jacobs-Reitsma, W.F.; Stegeman, A.; Klinkenberg, D.

    2009-01-01

    Since meat from poultry colonized with Campylobacter spp. is a major cause of bacterial gastroenteritis, human exposure should be reduced by, among other things, prevention of colonization of broiler flocks. To obtain more insight into possible sources of introduction of Campylobacter into broiler f

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

    Science.gov (United States)

    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...

  18. Isolation, identification and antibiotic resistance of Campylobacter strains isolated from domestic and free-living pigeons.

    Science.gov (United States)

    Dudzic, A; Urban-Chmiel, R; Stępień-Pyśniak, D; Dec, M; Puchalski, A; Wernicki, A

    2016-04-01

    1. The aim of this study was to evaluate the occurrence of Campylobacter spp. in domestic and free-living pigeons and to evaluate the antibiotic resistance profiles. 2. The material consisted of cloacal swabs obtained from 108 homing pigeons and fresh faeces from 72 wild birds from Lublin and its vicinity. The identification of strains isolated on differential/selective media for Campylobacter spp. was carried out by MALDI-TOF and PCR. The susceptibility to antibiotics was evaluated by minimum inhibitory concentration (MIC) in Mueller-Hinton broth. 3. A total of 35 strains of Campylobacter spp. were isolated; 27 were identified as Campylobacter jejuni and 8 as Campylobacter coli. Over half of the isolates were resistant to erythromycin and streptomycin, 40% of strains were resistant to tetracycline and ampicillin and 37% isolates were resistant to amoxicillin. Resistance to two or more antibiotics was observed in all strains tested. 4. The results indicate that both domestic and free-living pigeons are reservoirs for bacteria of the genus Campylobacter, which are characterised by varied and growing resistance to commonly used antibiotics. PMID:26841300

  19. The natural antimicrobial carvacrol inhibits Campylobacter jejuni motility and infection of epithelial cells.

    Directory of Open Access Journals (Sweden)

    Lieke B van Alphen

    Full Text Available BACKGROUND: Natural compounds with anti-microbial properties are attractive reagents to reduce the use of conventional antibiotics. Carvacrol, the main constituent of oregano oil, inhibits the growth of a variety of bacterial foodborne pathogens. As concentrations of carvacrol may vary in vivo or when used in animal feed, we here investigated the effect of subinhibitory concentrations of the compound on major virulence traits of the principal bacterial foodborne pathogen Campylobacter jejuni. METHODS/PRINCIPAL FINDINGS: Motility assays revealed that subinhibitory concentrations of carvacrol inhibited the motility of C. jejuni without affecting bacterial growth. Immunoblotting and electron microscopy showed that carvacrol-treated C. jejuni still expressed flagella. The loss of motility was not caused by reduced intracellular ATP levels. In vitro infection assays demonstrated that subinhibitory concentrations of carvacrol also abolished C. jejuni invasion of human epithelial cells. Bacterial uptake of invasive Escherichia coli was not blocked by carvacrol. Exposure of C. jejuni to carvacrol prior to infection also inhibited cellular infection, indicating that the inhibition of invasion was likely caused by an effect on the bacteria rather than inhibition of epithelial cell function. CONCLUSIONS/SIGNIFICANCE: Bacterial motility and invasion of eukaryotic cells are considered key steps in C. jejuni infection. Our results indicate that subinhibitory concentrations of carvacrol effectively block these virulence traits by interfering with flagella function without disturbing intracellular ATP levels. These results broaden the spectrum of anti-microbial activity of carvacrol and support the potential of the compound for use in novel infection prevention strategies.

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

    Science.gov (United States)

    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.

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

    Directory of Open Access Journals (Sweden)

    Jenny Olofsson

    2015-09-01

    Full Text Available 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.

  2. Re-annotation and re-analysis of the Campylobacter jejuni NCTC11168 genome sequence

    Directory of Open Access Journals (Sweden)

    Dorrell Nick

    2007-06-01

    Full Text Available Abstract Background Campylobacter jejuni is the leading bacterial cause of human gastroenteritis in the developed world. To improve our understanding of this important human pathogen, the C. jejuni NCTC11168 genome was sequenced and published in 2000. The original annotation was a milestone in Campylobacter research, but is outdated. We now describe the complete re-annotation and re-analysis of the C. jejuni NCTC11168 genome using current database information, novel tools and annotation techniques not used during the original annotation. Results Re-annotation was carried out using sequence database searches such as FASTA, along with programs such as TMHMM for additional support. The re-annotation also utilises sequence data from additional Campylobacter strains and species not available during the original annotation. Re-annotation was accompanied by a full literature search that was incorporated into the updated EMBL file [EMBL: AL111168]. The C. jejuni NCTC11168 re-annotation reduced the total number of coding sequences from 1654 to 1643, of which 90.0% have additional information regarding the identification of new motifs and/or relevant literature. Re-annotation has led to 18.2% of coding sequence product functions being revised. Conclusions Major updates were made to genes involved in the biosynthesis of important surface structures such as lipooligosaccharide, capsule and both O- and N-linked glycosylation. This re-annotation will be a key resource for Campylobacter research and will also provide a prototype for the re-annotation and re-interpretation of other bacterial genomes.

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

    OpenAIRE

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

    2014-01-01

    Although bacterial mechanisms involved in the resistance to inorganic arsenic are well understood, the molecular basis for organic arsenic resistance has not been described. Campylobacter jejuni, a major food-borne pathogen causing gastroenteritis in humans, is highly prevalent in poultry and is reportedly resistant to the arsenic compound roxarsone (4-hydroxy-3-nitrobenzenearsonic acid), which has been used as a feed additive in the poultry industry for growth promotion. In this study, we re...

  4. A case of myopericarditis associated to Campylobacter jejuni infection in the southern hemisphere

    OpenAIRE

    Alberto Fica; Daniela Seelmann; Lorena Porte; Daniela Eugenin; Ricardo Gallardo

    2012-01-01

    Myopericarditis is an infrequent complication of acute diarrheal illness due to Campylobacter jejuni, and it has been mainly reported in developed nations. The first case detected in Chile - an upper-middle income country -, that is coincidental with the increasing importance of acute gastroenteritis associated to this pathogen, is described. Recognition of this agent in stools requires special laboratory techniques not widely available, and it was suspected when a young patient presented wit...

  5. Identification and Analysis of Flagellar Co-expressed Determinants (Feds) of Campylobacter jejuni Involved in Colonization

    OpenAIRE

    Barrero-Tobon, Angelica M.; Hendrixson, David R.

    2012-01-01

    The flagellum of Campylobacter jejuni provides motility essential for commensal colonization of the intestinal tract of avian species and infection of humans resulting in diarrheal disease. Additionally, the flagellar type III secretion system has been reported to secrete proteins such as CiaI that influence invasion of human intestinal cells and possibly pathogenesis. The flagellar regulatory system ultimately influences σ28 activity required for expression of the FlaA major flagellin and ot...

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

    Energy Technology Data Exchange (ETDEWEB)

    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,

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

    Science.gov (United States)

    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) 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.

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

    Science.gov (United States)

    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.

  9. Basolateral invasion and trafficking of Campylobacter jejuni in polarized epithelial cells.

    Directory of Open Access Journals (Sweden)

    Lieneke I Bouwman

    Full Text Available Campylobacter jejuni is a major cause of bacterial diarrheal disease. Most enteropathogenic bacteria including C. jejuni can invade cultured eukaryotic cells via an actin- and/or microtubule-dependent and an energy-consuming uptake process. Recently, we identified a novel highly efficient C. jejuni invasion pathway that involves bacterial migration into the subcellular space of non-polarized epithelial cells (termed subvasion followed by invasion from the cell basis. Here we report cellular requirements of this entry mechanism and the subsequent intracellular trafficking route of C. jejuni in polarized islands of Caco-2 intestinal epithelial cells. Advanced microscopy on infected cells revealed that C. jejuni invades the polarized intestinal cells via the subcellular invasion pathway. Remarkably, invasion was not blocked by the inhibitors of microtubule dynamics colchicine or paclitaxel, and was even enhanced after disruption of host cell actin filaments by cytochalasin D. Invasion also continued after dinitrophenol-induced cellular depletion of ATP, whereas this compound effectively inhibited the uptake of invasive Escherichia coli. Confocal microscopy demonstrated that intracellular C. jejuni resided in membrane-bound CD63-positive cellular compartments for up to 24 h. Establishment of a novel luciferase reporter-based bacterial viability assay, developed to overcome the limitations of the classical bacterial recovery assay, demonstrated that a subset of C. jejuni survived intracellularly for up to 48 h. Taken together, our results indicate that C. jejuni is able to actively invade polarized intestinal epithelial cells via a novel actin- and microtubule-independent mechanism and remains metabolically active in the intracellular niche for up to 48 hours.

  10. [Campylobacter spp.: prevalence and pheno-genotypic characterization of isolates recovered from patients suffering from diarrhea and their pets in La Pampa Province, Argentina].

    Science.gov (United States)

    Tamborini, Ana L; Casabona, Luis M; Viñas, María R; Asato, Valeria; Hoffer, Alicia; Farace, María I; Lucero, María C; Corso, Alejandra; Pichel, Mariana

    2012-01-01

    The prevalence of Campylobacter spp. was investigated in 327 patients suffering from diarrhea and in 36 animals (dogs, cats and chickens) owned by the patients that presented infection by Campylobacter in Santa Rosa, La Pampa, Argentina. Campylobacter spp. was isolated in 50/327 patients and in 12/36 animals, being Campylobacter jejuni the most common species. Resistance to ciprofloxacin (65 %) and tetracycline (32 %) was found among 35 isolates of human origin studied. Seven genetic subtypes were observed among 13 C. jejuni isolates by pulsed field gel electrophoresis. Two subtypes grouped isolates belonging to patients and their respective dogs whereas another subtype grouped one isolate of human origin and two isolates from the patient's chickens. The results of this investigation highlight the need to strengthen surveillance of Campylobacter spp. not only in poultry, which is recognized as the main reservoir, but also in pets, which were shown to be asymptomatic carriers of the pathogen. PMID:23267623

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

    Science.gov (United States)

    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

  12. Use of reducing agents for the aerobic growth ofCampylobacter jejuni

    Institute of Scientific and Technical Information of China (English)

    Siddiqui R; Siti Asma H

    2010-01-01

    Objective:To produce a technique for the growth ofCampylobacter jejuni in aerobic condition Methods:Different combinations of reducing agents were tested in brucella broth and the growth turbidity was compared with tubes containing normal broth only. Microaerophilic environment was also provided in a petri plate seeded withCampylobacter culture by pouring 3 different concentrations (10%, 5% and 2.5%) of five reducing agents along with bacto-agar in the lid which was used to cover and seal the culture plate. Six reducing agents were also added in broth in concentration of 0.25 mg/mL of each with different combinations.Results: In lid agar technique,Campylobacter jejunigrowth appeared in all three concentrations of reducing agents, that is 10%, 5% and 2.5% after 24 hours of incubation but the best results were observed in 10% concentration. The colonial and morphological characters were not affected when the organisms were grown by this technique.Conclusions: It was found that reducing agents enhance the growth ofC. jejuni/coli. In combination of FeSO4, Na2CO3 with H3BO3 worked as ideal mixture for the aerobic growth ofCampylobacter. This technique is more economical as compared to commercially available media in the market and can be used for the oral facultative and microaerophilic bacterial growth in laboratory.

  13. Heterogeneity of Campylobacter species isolated from serial stool specimens of Egyptian children using pulsed field gel electrophoresis

    Directory of Open Access Journals (Sweden)

    Atef M. El-Gendy

    2013-03-01

    Full Text Available Background: The genus Campylobacter spp. is a common cause of human acute bacteria lenteritis and travellers’ diarrhoea worldwide.Objective: To determine whether multiple serial isolations of Campylobacter spp., when obtained from a single child, represented the same or a different organism.Methods: In a birth cohort study conducted in Egypt, numerous children showed serial isolations of Campylobacter spp. Of these, 13 children were selected from different households based on the successive isolation of six or more Campylobacter isolates from stool samples.Results: Eighty isolates were recovered and identified as either Campylobacter coli (n = 25 or Campylobacter jejuni (n = 55. Pulsed-field gel electrophoresis (PFGE revealed the presence of 38 unique C. jejuni and 24 C. coli profiles at a similarity level of ≥ 90%. Although no seriallyidentical isolates were detected in six children, others demonstrated at least one identical couple of isolates; all identified serially between one to six weeks. Two children demonstrated > 80% similar couples of isolates that appeared seven months apart. PFGE could be a useful tool for differentiating reinfection, relapse and convalescent excretion phases.Conclusion: Our data suggest that Campylobacter infection in children is a complex process; children are exposed to multiple species in endemic environments and strains of the same bacterium appear to be shed serially between one to six weeks after the first exposure. Isolates that persisted for longer periods were relatively less similar, as shown from the results of this study.

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

    Science.gov (United States)

    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

  15. Use of a Rabbit Soft Tissue Chamber Model to Investigate Campylobacter jejuni - Host Interactions

    Directory of Open Access Journals (Sweden)

    Annika eFlint

    2010-11-01

    Full Text Available Despite the prevalence of C. jejuni as an important food borne pathogen, the microbial factors governing its infection process are poorly characterized. In this study, we developed a novel rabbit soft tissue chamber model to investigate C. jejuni interactions with its host. The in vivo transcriptome profile of C. jejuni was monitored as a function of time post-infection by competitive microarray hybridization with cDNA obtained from C. jejuni grown in vitro. Genome-wide expression analysis identified 449 genes expressed at significantly different levels in vivo. Genes implicated to play important roles in early colonization of C. jejuni within the tissue chamber include up-regulation of genes involved in ribosomal protein synthesis and modification, heat shock response, and primary adaptation to the host environment (DccSR regulon. Genes encoding proteins involved in the TCA cycle and flagella related components were found to be significantly down regulated during early colonization. Oxidative stress defense and stringent response genes were found to be maximally induced during the acute infectious phase. Overall, these findings reveal possible mechanisms involved in adaptation of Campylobacter to the host.

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

    Science.gov (United States)

    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.

  17. Campylobacter jejuni cocultured with epithelial cells reduces surface capsular polysaccharide expression.

    LENUS (Irish Health Repository)

    Corcionivoschi, N

    2012-02-01

    The host cell environment can alter bacterial pathogenicity. We employed a combination of cellular and molecular techniques to study the expression of Campylobacter jejuni polysaccharides cocultured with HCT-8 epithelial cells. After two passages, the amount of membrane-bound high-molecular-weight polysaccharide was considerably reduced. Microarray profiling confirmed significant downregulation of capsular polysaccharide (CPS) locus genes. Experiments using conditioned media showed that sugar depletion occurred only when the bacterial and epithelial cells were cocultured. CPS depletion occurred when C. jejuni organisms were exposed to conditioned media from a different C. jejuni strain but not when exposed to conditioned media from other bacterial species. Proteinase K or heat treatment of conditioned media under coculture conditions abrogated the effect on the sugars, as did formaldehyde fixation and cycloheximide treatment of host cells or chloramphenicol treatment of the bacteria. However, sugar depletion was not affected in flagellar export (fliQ) and quorum-sensing (luxS) gene mutants. Passaged C. jejuni showed reduced invasiveness and increased serum sensitivity in vitro. C. jejuni alters its surface polysaccharides when cocultured with epithelial cells, suggesting the existence of a cross talk mechanism that modulates CPS expression during infection.

  18. Campy lobacter jejuni isolated from a patient with bacteremia in Guizhou Province, China%贵州省首例空肠弯曲菌菌血症病例病原鉴定和亚种分型

    Institute of Scientific and Technical Information of China (English)

    韦小瑜; 田克诚; 游旅; 唐光鹏; 王定明

    2014-01-01

    目的:对贵州省1例菌血症患儿血液中分离的疑似空肠弯曲菌进行鉴定。方法运用传统细菌学方法和分子生物学方法,对从菌血症患者血液分离的可疑空肠弯曲菌进行鉴定和亚种分型。结果来自菌血症患儿血液的可疑菌株,经传统生化鉴定为空肠弯曲菌空肠亚种,特异性多重PCR方法鉴定为弯曲菌属空肠弯曲菌,NAP-mPCR方法鉴定为空肠弯曲菌空肠亚种。结论分离自贵州省菌血症患儿血液的菌株确认为空肠弯曲菌空肠亚种,NAP-mPCR方法可将空肠弯曲菌鉴定到亚种水平。%To identify the isolated suspicious strain of Campylobacter jejuni from the blood of bacteremia patient in Guizhou Province ,China ,conventional and molecular techniques (specific mPCR and NAP-mPCR) were used to identify suspi-cious bacteria strains .Results showed that Campylobacter jejuni suspicious colonies were cultured in bacteremia patient blood samples .The strain was identified as Campylobacter jejuni ssp . jejuni by conventional tests and was identified as Campy-lobacter jejuni by genus specific mPCR .Then the strain was classified as Campylobacter jejuni ssp . jejuni by subspecies NAP-mPCR .The strain was identified as Campylobacter jejuni ssp .jejuni isolated from the blood of bacteremia patient and Campylobacter jejuni can be identified subspecies by NAP-mPCR .

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

    Directory of Open Access Journals (Sweden)

    Nabila eHaddad

    2012-03-01

    Full Text Available Polynucleotide phosphorylase, 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 the two major ribonucleases (RNase E and RNase II of Escherichia coli are absent from Campylobacter genome, this study was focused on 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 (NANA 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.

  20. The complete Campylobacter jejuni transcriptome during colonization of a natural host determined by RNAseq.

    Directory of Open Access Journals (Sweden)

    Michael E Taveirne

    Full Text Available Campylobacter jejuni is a major human pathogen and a leading cause of bacterial derived gastroenteritis worldwide. C. jejuni regulates gene expression under various environmental conditions and stresses, indicative of its ability to survive in diverse niches. Despite this ability to highly regulate gene transcription, C. jejuni encodes few transcription factors and its genome lacks many canonical transcriptional regulators. High throughput deep sequencing of mRNA transcripts (termed RNAseq has been used to study the transcriptome of many different organisms, including C. jejuni; however, this technology has yet to be applied to defining the transcriptome of C. jejuni during in vivo colonization of its natural host, the chicken. In addition to its use in profiling the abundance of annotated genes, RNAseq is a powerful tool for identifying and quantifying, as-of-yet, unknown transcripts including non-coding regulatory RNAs, 5' untranslated regulatory elements, and anti-sense transcripts. Here we report the complete transcriptome of C. jejuni during colonization of the chicken cecum and in two different in vitro growth phases using strand-specific RNAseq. Through this study, we identified over 250 genes differentially expressed in vivo in addition to numerous putative regulatory RNAs, including trans-acting non-coding RNAs and anti-sense transcripts. These latter potential regulatory elements were not identified in two prior studies using ORF-based microarrays, highlighting the power and value of the RNAseq approach. Our results provide new insights into how C. jejuni responds and adapts to the cecal environment and reveals new functions involved in colonization of its natural host.

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

    Science.gov (United States)

    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.

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

    Science.gov (United States)

    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

  3. Regulation of oxidative stress response by CosR, an essential response regulator in Campylobacter jejuni.

    Directory of Open Access Journals (Sweden)

    Sunyoung Hwang

    Full Text Available CosR (Campylobacter oxidative stress regulator; Cj0355c is an OmpR-type response regulator essential for the viability of Campylobacter jejuni, a leading foodborne pathogen causing human gastroenteritis worldwide. Despite importance, the function of CosR remains completely unknown mainly because of cell death caused by its knockout mutation. To overcome this technical limitation, in this study, antisense technology was used to investigate the regulatory function of CosR by modulating the level of CosR expression. Two-dimensional gel electrophoresis (2DGE was performed to identify the CosR regulon either by suppressing CosR expression with antisense peptide nucleic acid (PNA or by overexpressing CosR in C. jejuni. According to the results of 2DGE, CosR regulated 32 proteins involved in various cellular processes. Notably, CosR negatively regulated a few key proteins of the oxidative stress response of C. jejuni, such as SodB, Dps, Rrc and LuxS, whereas CosR positively controlled AhpC. Electrophoretic mobility shift assay showed that CosR directly bound to the promoter region of the oxidative stress genes. DNase I footprinting assays identified 21-bp CosR binding sequences in the sodB and ahpC promoters, suggesting CosR specifically recognizes and binds to the regulated genes. Interestingly, the level of CosR protein was significantly reduced by paraquat (a superoxide generator but not by hydrogen peroxide. Consistent with the overall negative regulation of oxidative stress defense proteins by CosR, the CosR knockdown by antisense rendered C. jejuni more resistant to oxidative stress compared to the wild type. Overall, this study reveals the important role played by the essential response regulator CosR in the oxidative stress defense of C. jejuni.

  4. Variation of chemosensory receptor content of Campylobacter jejuni strains and modulation of receptor gene expression under different in vivo and in vitro growth conditions

    Directory of Open Access Journals (Sweden)

    Day Christopher J

    2012-06-01

    Full Text Available Abstract Background Chemotaxis is crucial for the colonisation/infection of hosts with Campylobacter jejuni. Central to chemotaxis are the group A chemotaxis genes that are responsible for sensing the external environment. The distribution of group A chemoreceptor genes, as found in the C. jejuni sequenced strains, tlp1-4, 7, 10 and 11 were determined in 33 clinical human and avian isolates. Results Group A tlp gene content varied among the strains with genes encoding tlp1 (aspartate receptor, ccaA and tlp7 present in all strains tested, where as tlp11 was present in only one of our international collection clinical isolates, C. jejuni 520, but was more prevalent (9/13 in the freshly isolated clinical stains from patients who required hospitalisation due to C. jejuni infection (GCH1-17. Relative expression levels of the group A tlp genes were also determined in C. jejuni reference strains NCTC 11168-GS, 11168-O and 81116 using cells grown in vitro at 37°C, 42°C and maintained at room temperature and with cells isolated directly from murine and avian hosts by immune magnetic separation without subsequent culture. Gene expression of tlp genes was varied based on strain, growth conditions and in vivo isolation source. Tlp1, although the most conserved, showed the lowest and most varied mRNA expression and protein production under laboratory conditions. Tlp7 was highly expressed at most conditions tested, and gene expression was not influenced by the tlp7 gene encoding a full length protein or one expressed as separate periplasmic and cytoplasmic domains. Conclusion We have shown that chemosensory receptor set variation exists among C. jejuni strains, but is not dependent on the isolation source.

  5. Campylobacter iguaniorum sp. nov., isolated from reptiles

    Science.gov (United States)

    During samplings of reptiles for Epsilonproteobacteria, Campylobacter strains were isolated from lizards and chelonians not belonging to any of the established taxa. Initial AFLP, PCR, and 16S rRNA sequence analysis showed that these strains were most closely related to Campylobacter fetus and Campy...

  6. Effect of environmental stress factors on the uptake and survival of Campylobacter jejuni in Acanthamoeba castellanii

    DEFF Research Database (Denmark)

    Bui, Thanh Xuan; Qvortrup, Klaus; Wolff, Anders;

    2012-01-01

    Background: Campylobacter jejuni is a major cause of bacterial food-borne illness in Europe and North America. The mechanisms allowing survival in the environment and transmission to new hosts are not well understood. Environmental free-living protozoa may facilitate both processes. Pre-exposure ......Background: Campylobacter jejuni is a major cause of bacterial food-borne illness in Europe and North America. The mechanisms allowing survival in the environment and transmission to new hosts are not well understood. Environmental free-living protozoa may facilitate both processes. Pre...... was slightly up-regulated under heat and oxidative stresses but down-regulated under starvation and osmotic stresses, the htrA gene showing the largest down-regulation in response to osmotic stress. Pre-exposure of bacteria to low nutrient or osmotic stress reduced bacterial uptake by amoeba, but no effect...... of heat or oxidative stress was observed. Finally, C. jejuni rapidly lost viability within amoeba cells and pre-exposure to oxidative stress had no significant effect on intracellular survival. However, the numbers of intracellular bacteria recovered 5 h post-gentamicin treatment were lower with starved...

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

    Science.gov (United States)

    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 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

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

    Science.gov (United States)

    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

  9. Molecular Mimicry: Sensitization of Lewis Rats With Campylobacter jejuni Lipopolysaccharides Induces Formation of Antibody Toward GD3 Ganglioside

    OpenAIRE

    Usuki, Seigo; Thompson, Stuart A.; Rivner, Michael H.; Taguchi, Kyoji; Shibata, Keiko; Ariga, Toshio; Robert K Yu

    2006-01-01

    Recently we have reported cases of demyelinating inflammatory neuropathy showing elevated titers of anti-GD3 antibodies, which occurs rarely in Guillain-Barré syndrome. To examine the correlation between the anti-GD3 antibody titer and Campylobacter jejuni infection, we sensitized female Lewis rats with lipopolysaccharides (LPSs) from serotype HS19 of C. jejuni and examined changes in nerve conduction velocity and nerve conduction block (P/D ratio). After 16 weeks of sensitization, animals re...

  10. Characterization of lipooligosaccharide-biosynthetic loci of Campylobacter jejuni reveals new lipooligosaccharide classes: Evidence of mosaic organizations

    OpenAIRE

    Parker, Craig; Gilbert, Michel; Yuki, Nobuhiro; Endtz, Hubert; Mandrell, Robert

    2008-01-01

    textabstractThe lipooligosaccharide (LOS) biosynthesis region is one of the more variable genomic regions between strains of Campylobacter jejuni. Indeed, eight classes of LOS biosynthesis loci have been established previously based on gene content and organization. In this study, we characterize additional classes of LOS biosynthesis loci and analyze various mechanisms that result in changes to LOS structures. To gain further insights into the genomic diversity of C. jejuni LOS biosynthesis ...

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

    Science.gov (United States)

    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.

  12. Crystal structure of the Campylobacter jejuni Cj0090 protein reveals a novel variant of the immunoglobulin fold among bacterial lipoproteins

    OpenAIRE

    Paek, Seonghee; Kawai, Fumihiro; Choi, Kyoung-Jae; Yeo, Hye-Jeong

    2012-01-01

    Bacterial lipoproteins play an important role in bacterial pathogenesis and physiology. The genome of Campylobacter jejuni, a major foodborn pathogen, is predicted to contain over 20 lipoproteins. However, the functions of the majority of C. jejuni lipoproteins remain unknown. The Cj0090 protein is encoded by a lipoprotein operon composed of cj0089, cj0090, and cj0091. Here, we report the crystal structure of Cj0090 at 1.9 Å resolution, revealing a novel variant of the immunoglobulin fold wit...

  13. Campylobacter jejuni infection increases anxiety-like behavior in the holeboard: possible anatomical substrates for viscerosensory modulation of exploratory behavior

    OpenAIRE

    Goehler, Lisa E.; Park, Su Mi; Opitz, Noel; Lyte, Mark; Gaykema, Ronald P.A.

    2007-01-01

    The presence of certain bacteria in the gastrointestinal tract influences behavior and brain function. For example, challenge with live Campylobacter jejuni (C. jejuni), a common food-born pathogen, reduces exploration of open arms of the plus maze, consistent with anxiety-like behavior, and activates brain regions associated with autonomic function, likely via a vagal pathway. As yet, however, little is known regarding the interface of immune sensory signals with brain substrates that mediat...

  14. 空肠弯曲杆菌ERIC-PCR分型技术研究%Study on construction and application of ERIC-PCR method for typing Campylobacter jejuni

    Institute of Scientific and Technical Information of China (English)

    唐梦君; 高玉时; 张小燕; 陆俊贤; 施祖灏; 唐修君; 陈大伟

    2013-01-01

    目的:建立空肠弯曲杆菌肠杆菌科基因间重复一致序列PCR(ERIC-PCR)的分子分型技术,快速分析空肠弯曲杆菌指纹谱。方法本研究提取空肠弯曲杆菌ATCC33560的基因组DNA作为模板,采用对优化项目设置梯度其它条件不变的方法,对ERIC-PCR反应中的模板量、引物浓度和退火温度等进行优化,建立空肠弯曲杆菌的ERIC-PCR分型技术,并采用该技术对20株空肠弯曲杆菌的指纹图谱进行了分析。结果 ERIC-PCR反应体系中DNA模板量96ng/25μL,ERIC1、ERIC2引物浓度各0.8μmol/L,退火温度40℃时,指纹图谱条带清晰、明亮;20株空肠弯曲杆菌临床分离株ERIC-PCR指纹图谱差异较大,可在160bp~32000bp范围内出现2条~13条条带,分为14型。结论研究显示成功建立了空肠弯曲杆菌ERIC-PCR指纹图谱分型技术,应用该方法可从分子水平上对空肠弯曲杆菌基因组DNA进行快速指纹图谱分析,具有简便和快速等优点,能够为空肠弯曲杆菌流行病学调查提供科学依据。%Objective To generate an efifcient enterobacterial repetitive intergenic consensus sequence (ERIC-PCR) typing and analyze molecular type of different food-borne Campylobacter jejuni.Method Genomic DNA of Campylo-bacter jejuni reference strain ATCC33560 was used as the template for PCR. Target sequences in Campylobacter jejuni genomic DNA were amplified with the primers designed according to the reference. To obtain optimum fingerprint maps,template and primer concentration of the reaction system and annealing temperature of PCR,the factors to be optimized were designed in different concentration grads and other factors were ifxed. Then 20 Campylobacter jejuni isolates were classiifed by PCR.Result The optimal concentration of template DNA was 96ng/25mL,the concentration of primers was 0.8mmol/L each primer and annealing temperature of PCR was 40 ℃.The ERIC-PCR ifngerprint maps of different isolates

  15. Interleukin-18 Mediates Immune Responses to Campylobacter jejuni Infection in Gnotobiotic Mice.

    Directory of Open Access Journals (Sweden)

    Stefan Bereswill

    Full Text Available Human Campylobacter jejuni infections are progressively rising worldwide. Information about the molecular mechanisms underlying campylobacteriosis, however, are limited. In the present study we investigated whether cytokines such as IL-23, IL-22 and IL-18, which share pivotal functions in host immunity, were involved in mediating intestinal and systemic immunopathological responses upon C. jejuni infection.To assure stable infection, gnotobiotic (i.e. secondary abiotic IL-23p19-/-, IL-22-/- and IL-18-/- mice were generated by broad-spectrum antibiotic treatment. Following peroral C. jejuni strain 81-176 infection, mice of all genotypes harbored comparably high pathogenic loads in their intestines. As compared to wildtype controls, however, IL-18-/- mice displayed less distinct C. jejuni induced sequelae as indicated by less pronounced large intestinal shrinkage and lower numbers of apoptotic cells in the colonic epithelial layer at day 8 postinfection (p.i.. Furthermore, lower colonic numbers of adaptive immune cells including regulatory T cells and B lymphocytes were accompanied by less distinct secretion of pro-inflammatory cytokines such as TNF and IFN-γ and lower IL-17A mRNA expression levels in colonic ex vivo biopsies of infected IL-18-/- as compared to wildtype mice. Upon C. jejuni infection, colonic IL-23p19 expression was up-regulated in IL-18-/- mice only, whereas IL-22 mRNA levels were lower in uninfected and infected IL-23p19-/- as well as infected IL-18-/- as compared to respective wildtype control mice. Remarkably, not only intestinal, but also systemic infection-induced immune responses were less pronounced in IL-18-/- mice as indicated by lower TNF, IFN-γ and IL-6 serum levels as compared to wildtype mice.We here show for the first time that IL-18 is essentially involved in mediating C. jejuni infection in the gnotobiotic mouse model. Future studies need to further unravel the underlying regulatory mechanisms orchestrating

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

    Directory of Open Access Journals (Sweden)

    van Bunnik Bram AD

    2012-07-01

    Full Text Available Abstract Background Infectious diseases in plants, animals and humans are often transmitted indirectly between hosts (or between groups of hosts, i.e. via some route through the environment instead of via direct contacts between these hosts. Here we study indirect transmission experimentally, using transmission of Campylobacter jejuni (C. jejuni between spatially separated broilers as a model system. We distinguish three stages in the process of indirect transmission; (1 an infectious “sender” excretes the agent, after which (2 the agent is transported via some route to a susceptible “receiver”, and subsequently (3 the receiver becomes colonised by the agent. The role of the sender and receiver side (stage 1 and stage 3 was studied here by using acidification of the drinking water as a modulation mechanism. Results In the experiment one control group and three treatment groups were monitored for the presence of C. jejuni by taking daily cloacal swabs. The three treatments consisted of acidification of the drinking water of the inoculated animals (the senders, acidification of the drinking water of the susceptible animals (the receivers or acidification of the drinking water of both inoculated and susceptible animals. In the control group 12 animals got colonised out of a possible 40, in each treatment groups 3 animals out of a possible 40 were found colonised with C. jejuni. Conclusions The results of the experiments show a significant decrease in transmission rate (β between the control groups and treatment groups (p

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

    Science.gov (United States)

    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

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

    Science.gov (United States)

    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.

  19. Global gene expression analysis of chicken caecal response to Campylobacter jejuni.

    Science.gov (United States)

    Shaughnessy, Ronan G; Meade, Kieran G; McGivney, Beatrice A; Allan, Brenda; O'Farrelly, Cliona

    2011-07-15

    Campylobacter jejuni colonises the caecum of more than 90% of commercial chickens. Even though colonisation is asymptomatic, we hypothesised that it is mediated by activation of several biological pathways. We therefore used chicken-specific 20K oligonucleotide microarrays to examine global gene expression in C. jejuni-challenged birds. Microarray results demonstrate small but significant fold-changes in expression of 270 genes 20 h post-challenge, corresponding to a wide range of biological processes including cell growth, nutrient metabolism and immunological activity. Expression of NOX1 (2.3-fold) and VCAM1 (1.5-fold) were significantly increased in colonised birds (PCTLA4 (2.5-fold) gene expression (Pintestinal T cell response. PMID:21605915

  20. Different contributions of HtrA protease and chaperone activities to Campylobacter jejuni stress tolerance and physiology

    DEFF Research Database (Denmark)

    Bæk, Kristoffer Torbjørn; Vegge, Christina Skovgaard; Skórko-Glonek, Joanna;

    2011-01-01

    The microaerophilic bacterium Campylobacter jejuni is the most common cause of bacterial food-borne infections in the developed world. Tolerance to environmental stress relies on proteases and chaperones in the cell envelope such as HtrA and SurA. HtrA displays both chaperone and protease activity......, but little is known about how each of these activities contributes to stress tolerance in bacteria. In vitro experiments showed temperature dependent protease and chaperone activities of C. jejuni HtrA. A C. jejuni mutant lacking only the protease activity of HtrA was used to show that the HtrA chaperone...

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

    Directory of Open Access Journals (Sweden)

    Naikare Hemant

    2009-10-01

    Full Text Available Abstract 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.

  2. Hygromycin B and apramycin antibiotic resistance cassettes for use in Campylobacter jejuni.

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    Andrew Cameron

    Full Text Available 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.

  3. A PAS Domain-Containing Regulator Controls Flagella-Flagella Interactions in Campylobacter jejuni

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    Mark eReuter

    2015-07-01

    Full Text Available The bipolar flagella of the foodborne bacterial pathogen Campylobacter jejuni confermotility, which is essential for virulence. The flagella of C. jejuni are posttranslationallymodified, but how this process is controlled is not well understood. Inthis work, we have identified a novel PAS-domain containing regulatory system, whichmodulates 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 thegeneration of a tightly associated 'cell-train' morphotype, where up to four cells wereconnected 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 genomicsshowed that cj1387c (yheO orthologs in bacterial genomes are commonly linked to anadjacent cj1388 ortholog, with some bacteria, including C. jejuni, containing anothercj1388-like gene (cj0327. Inactivation of the cj1388 and cj0327 genes resulted indecreased autoagglutination in Tween-20-supplemented media. The Δcj1388 andΔcj0327 mutants were also attenuated in a Galleria larvae-based infection model.Finally, substituting the sole cysteine in Cj1388 for serine prevented Cj1388dimerisation in non-reducing conditions, and resulted in decreased autoagglutination inthe presence of Tween-20. We hypothesize that Cj1388 and Cj0327 modulate posttranslationalmodification of the flagella through yet unidentified mechanisms, andpropose naming Cj1387 the Campylobacter Flagella Interaction Regulator CfiR, andthe Cj1388 and Cj0327 protein as CfiP and CfiQ, respectively.

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

    Science.gov (United States)

    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

  5. Survival of Campylobacter jejuni in co-culture with Acanthamoeba castellanii: role of amoeba-mediated depletion of dissolved oxygen

    DEFF Research Database (Denmark)

    Bui, Thanh Xuan; Winding, Anne; Qvortrup, Klaus;

    2012-01-01

    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......, 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...

  6. Cepas de Campylobacter jejuni resistentes a quinolonas aisladas de humanos, gallinas y pollos

    OpenAIRE

    Rodolfo Notario; Noemí Borda; Telma Gambandé; Joaquín Bermejo; Adriana Ponessa; Virginia Toledo

    2011-01-01

    Se compararon 8 aislamientos de Campylobacter jejuni provenientes de humanos con enfermedad diarreica aguda, con 23 aislamientos de cloaca de gallinas y pollos obtenidos de zonas próximas a la ciudad de Rosario, todos resistentes a la ciprofloxacina. Las muestras se sembraron en agar selectivo y se incubaron en microaerofilia a 42 °C. Las colonias se identificaron con el método tradicional. Los aislamientos se conservaron a -70 °C en caldo cerebro corazón con 17% v/v de glicerina. La clonalid...

  7. Genetic diversity and antimicrobial resistance of Campylobacter and Salmonella strains isolated from decoys and raptors.

    Science.gov (United States)

    Jurado-Tarifa, E; Torralbo, A; Borge, C; Cerdà-Cuéllar, M; Ayats, T; Carbonero, A; García-Bocanegra, I

    2016-10-01

    Infections caused by thermotolerant Campylobacter spp. and Salmonella spp. are the leading causes of human gastroenteritis worldwide. Wild birds can act as reservoirs of both pathogens. A survey was carried out to determine the prevalence, genetic diversity and antimicrobial resistance of thermotolerant Campylobacter and Salmonella in waterfowl used as decoys and wild raptors in Andalusia (Southern Spain). The overall prevalence detected for Campylobacter was 5.9% (18/306; CI95%: 3.25-8.52) in decoys and 2.3% (9/387; CI95%: 0.82-3.83) in wild raptors. Isolates were identified as C. jejuni, C. coli and C. lari in both bird groups. Salmonella was isolated in 3.3% (10/306; CI95%: 2.3-4.3) and 4.6% (18/394; CI95%: 3.5-5.6) of the decoys and raptors, respectively. Salmonella Enteritidis and Typhimurium were the most frequently identified serovars, although Salmonella serovars Anatum, Bredeney, London and Mikawasima were also isolated. Pulsed-field gel electrophoresis analysis of isolates showed higher genetic diversity within Campylobacter species compared to Salmonella serovars. Campylobacter isolates showed resistance to gentamicin, ciprofloxacin and tetracycline, while resistance to erythromycin and tetracycline was found in Salmonella isolates. The results indicate that both decoys and raptors can act as natural carriers of Campylobacter and Salmonella in Spain, which may have important implications for public and animal health. PMID:27638115

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

    DEFF Research Database (Denmark)

    Gill, Carson; Bahrndorff, Simon; Lowenberger, Carl

    2016-01-01

    The house fly, Musca domestica, has been implicated as a vector of Campylobacter spp., a major cause of human disease. Little is known whether house flies serve as biological amplifying hosts or mechanical vectors for Campylobacter jejuni. We investigated the period after C. jejuni had been...... after ingestion but no countable C. jejuni colonies were observed > 24 hours post-ingestion. We detected viable C. jejuni in house fly vomitus and excreta up to 4 hours after ingestion, but no viable bacteria were detected ≥ 8 hours. Suppression subtractive hybridization identified pathogen-induced gene...

  9. Intestinal microbiota shifts towards elevated commensal Escherichia coli loads abrogate colonization resistance against Campylobacter jejuni in mice.

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    Lea-Maxie Haag

    Full Text Available BACKGROUND: The zoonotic pathogen Campylobacter jejuni is a leading cause of bacterial foodborne enterocolitis in humans worldwide. The understanding of immunopathology underlying human campylobacteriosis is hampered by the fact that mice display strong colonization resistance against the pathogen due to their host specific gut microbiota composition. METHODOLOGY/PRINCIPAL FINDINGS: Since the microbiota composition changes significantly during intestinal inflammation we dissected factors contributing to colonization resistance against C. jejuni in murine ileitis, colitis and in infant mice. In contrast to healthy animals C. jejuni could stably colonize mice suffering from intestinal inflammation. Strikingly, in mice with Toxoplasma gondii-induced acute ileitis, C. jejuni disseminated to mesenteric lymphnodes, spleen, liver, kidney, and blood. In infant mice C. jejuni infection induced enterocolitis. Mice suffering from intestinal inflammation and C. jejuni susceptible infant mice displayed characteristical microbiota shifts dominated by increased numbers of commensal Escherichia coli. To further dissect the pivotal role of those distinct microbiota shifts in abrogating colonization resistance, we investigated C. jejuni infection in healthy adult mice in which the microbiota was artificially modified by feeding live commensal E. coli. Strikingly, in animals harboring supra-physiological intestinal E. coli loads, colonization resistance was significantly diminished and C. jejuni infection induced enterocolitis mimicking key features of human campylobacteriosis. CONCLUSION/SIGNIFICANCE: Murine colonization resistance against C. jejuni is abrogated by changes in the microbiota composition towards elevated E. coli loads during intestinal inflammation as well as in infant mice. Intestinal inflammation and microbiota shifts thus represent potential risk factors for C. jejuni infection. Corresponding interplays between C. jejuni and microbiota might

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

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    Alexandre Thibodeau

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

  11. 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.

    Science.gov (United States)

    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

  12. Method Comparison for Enhanced Recovery, Isolation and Qualitative Detection of C. jejuni and C. coli from Wastewater Effluent Samples

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    María Ugarte-Ruiz

    2015-03-01

    Full Text Available Seeking a sensitive protocol, culture-dependent methods were compared to detect thermophilic Campylobacter species in untreated urban effluents. We evaluated various combinations of selective media, with and without an enrichment steps, as well as an extra filtration step. Culture-independent real-time quantitative PCR was also included and all detected isolates underwent antimicrobial susceptibility testing. All tested water samples contained Campylobacter DNA, but only 64% were positive after culture. Although enrichment using Preston broth resulted in better recovery of potentially stressed Campylobacter than Bolton or Campyfood broth (CFB, there was no significant increase in efficiency compared to direct plating. The type of selective agar media used, on the other hand, had a significant effect, with CASA plates performing better than mCCDA or CFA ones. Inclusion of an enrichment step increased the ratio of C. coli vs. C. jejuni being isolated. Resistances against all antimicrobials tested were observed in C. coli, but fewer instances of resistance were found in C. jejuni isolates. Whether this difference was the result of selection during the enrichment step could not be determined. The presence of Campylobacter in urban effluents can be considered as a valuable proxy for Campylobacter populations present in urban environments.

  13. Method comparison for enhanced recovery, isolation and qualitative detection of C. jejuni and C. coli from wastewater effluent samples.

    Science.gov (United States)

    Ugarte-Ruiz, María; Florez-Cuadrado, Diego; Wassenaar, Trudy M; Porrero, María Concepción; Domínguez, Lucas

    2015-03-02

    Seeking a sensitive protocol, culture-dependent methods were compared to detect thermophilic Campylobacter species in untreated urban effluents. We evaluated various combinations of selective media, with and without an enrichment steps, as well as an extra filtration step. Culture-independent real-time quantitative PCR was also included and all detected isolates underwent antimicrobial susceptibility testing. All tested water samples contained Campylobacter DNA, but only 64% were positive after culture. Although enrichment using Preston broth resulted in better recovery of potentially stressed Campylobacter than Bolton or Campyfood broth (CFB), there was no significant increase in efficiency compared to direct plating. The type of selective agar media used, on the other hand, had a significant effect, with CASA plates performing better than mCCDA or CFA ones. Inclusion of an enrichment step increased the ratio of C. coli vs. C. jejuni being isolated. Resistances against all antimicrobials tested were observed in C. coli, but fewer instances of resistance were found in C. jejuni isolates. Whether this difference was the result of selection during the enrichment step could not be determined. The presence of Campylobacter in urban effluents can be considered as a valuable proxy for Campylobacter populations present in urban environments.

  14. Polyphosphate kinase 2: a novel determinant of stress responses and pathogenesis in Campylobacter jejuni.

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    Dharanesh Gangaiah

    Full Text Available 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 Deltappk2 mutant exhibited a significant survival defect under osmotic, nutrient, aerobic, and antimicrobial stresses and displayed an enhanced ability to form static biofilms. However, the Deltappk2 mutant was not defective in poly P and ppGpp synthesis suggesting that PPK2-mediated stress tolerance is not ppGpp-mediated. Importantly, the Deltappk2 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.

  15. Prevalence and diversity of Campylobacter jejuni in pig herds on farms with and without cattle or poultry

    DEFF Research Database (Denmark)

    Boes, J.; Nersting, L.; Nielsen, Eva;

    2005-01-01

    The prevalence and diversity of Campylobacter jejuni was investigated in pig herds on farms with and without cattle or poultry production. A bacteriological screening of pig cecal samples from 247 finisher herds was carried out at the slaughterhouse. Subsequently, a follow-up study was conducted ...

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

    DEFF Research Database (Denmark)

    de Jong, Aarieke E I; van Asselt, Esther D; Zwietering, Marcel H;

    2012-01-01

    The aim of this research was to determine the decimal reduction times of bacteria present on chicken fillet in boiling water. The experiments were conducted with Campylobacter jejuni, Salmonella, and Escherichia coli. Whole chicken breast fillets were inoculated with the pathogens, stored overnight...

  17. Pyruvate relieves the necessity of high induction levels of catalase and enables Campylobacter jejuni to grow under fully aerobic conditions

    NARCIS (Netherlands)

    Verhoeff-Bakkenes, L.; Arends, A.P.; Snoep, J.L.; Zwietering, M.H.; Jonge, de R.

    2008-01-01

    Aims: Several cases of campylobacteriosis reported worldwide seemingly conflict with the strict growth requirements and sensitivity to environmental stress of Campylobacter jejuni. In this study, the need for a micro-aerobic environment [dissolved oxygen tension (DOT): 0·1¿90%; 100% air saturation)

  18. Quantification of Campylobacter jejuni cross-contamination via hands, cutlery, and cutting board during preparation of a chicken fruit salad

    NARCIS (Netherlands)

    Verhoeff-Bakkenes, L.; Beumer, R.R.; Jonge, de R.; Leusden, van F.M.; Jong, de A.E.I.

    2008-01-01

    Using artificially contaminated chicken, the quantitative overall effect of Campylobacter jejuni cross-contamination, either via cutlery, cutting board, or hands, on the microbiological quality of a chicken salad was tested to identify the most critical transfer route. The end contamination level of

  19. Characterization of lipooligosaccharide-biosynthetic loci of Campylobacter jejuni reveals new lipooligosaccharide classes: Evidence of mosaic organizations

    NARCIS (Netherlands)

    C.T. Parker (Craig); M. Gilbert (Michel); N. Yuki (Nobuhiro); H.P. Endtz (Hubert); R.E. Mandrell (Robert)

    2008-01-01

    textabstractThe lipooligosaccharide (LOS) biosynthesis region is one of the more variable genomic regions between strains of Campylobacter jejuni. Indeed, eight classes of LOS biosynthesis loci have been established previously based on gene content and organization. In this study, we characterize ad

  20. Control of VTEC O157 and Campylobacter jejuni/coli on cattle farms : Effective interventions and implementation

    NARCIS (Netherlands)

    Ellis-Iversen, J

    2009-01-01

    Verocytotoxogenic E. coli O157 (VTEC O157) and Campylobacter jejuni/coli are zoonotic pathogens of public health importance, which are commonly carried and shed by cattle. Control at farm level needed isto limit shedding and contamination of the environment and the human food chain. On- farm risk fa

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

    NARCIS (Netherlands)

    Jong, de A.E.I.; Asselt, van E.D.; Zwietering, M.H.; Nauta, M.J.; Jonge, de R.

    2012-01-01

    The aim of this research was to determine the decimal reduction times of bacteria present on chicken fillet in boiling water. The experiments were conducted with Campylobacter jejuni, Salmonella, and Escherichia coli. Whole chicken breast fillets were inoculated with the pathogens, stored overnight

  2. 用微量铁盐培养基分离空肠弯曲菌的效果研究%Effect of trace ferric salt medium in separation of campylobacter jejuni

    Institute of Scientific and Technical Information of China (English)

    李宏科

    2013-01-01

    Objective. -To explore the effect of trace ferric salt medium in separation of Campylobacter jejuni, and find the optimal culture medium for Campylobacter jejuni. Methods: The experiment was carried out under the same condition, Campylobacter jejuni were isolated and cultured with five different culture mediums. The growth of the bacteria was observed, the comparative experiment research was performed by colony counting method. Results: After comparing these five mediums, it was found the detection rate of trace ferric salt medium was remarkably higher than other culture medium in separation of Campylobacter jejuni, but there was also omission, in order to improve the detection rate, Campylobacter jejuni should be based on the simultaneous usage of the brucella blood AGAR and trace ferric salt medium. Conclusion: Trace ferric salt medium can be used for the disease epidemiological survey and clinical inspection of Campylobacter jejuni. The Campylobacter jejuni grows rapidly with large bacterial colony in large quantities on ferric salt medium, which is suggested to apply in laboratories for its low cost and easy preparation.%目的:探索用微量铁盐培养基分离空肠弯曲菌,寻求分离空肠弯曲菌良好的培养基.方法:采用五种不同培养基,在相同条件下对空肠弯曲菌分离培养,观察其生长效果,用菌落计数法进行比较试验.结果:通过比较,发现含微量铁盐培养基对空肠弯曲菌分离效果好,检出率高.但也有遗漏,为了提高检出率,应与布氏菌血琼脂同时使用.结论:含微量铁盐培养基可用于空肠弯曲菌病流行病学调查和临床检验,空肠弯曲菌在该培养基上生长较快,菌落大,数量多,且制备简易、价廉,建议各实验室推广应用.

  3. Prevalence and Antimicrobial Resistance of Thermophilic Campylobacter Isolated from Chicken in Côte d’Ivoire

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    Goualié Gblossi Bernadette

    2012-01-01

    Full Text Available Thermophilic Campylobacters are major causes of gastroenteritis in human. The main risk factor of infection is consumption of contaminated or by cross-contaminated poultry meat. In Côte d’Ivoire, gastroenteritis is usually observed but no case of human campylobacteriosis has been formally reported to date. The aims of this study were to determine prevalence and antimicrobial resistance of Campylobacter jejuni and Campylobacter coli isolated from chickens ceaca in commercial slaughter in Abidjan. Between May and November 2009, one hundred and nineteen (119 chicken caeca samples were collected and analyzed by passive filtration method followed by molecular identification (PCR. From these 119 samples, 76 (63.8% were positive to Campylobacter tests. Among the positive colonies, 51.3% were C. jejuni and 48.7% were C. coli. Of the 39 C. jejuni isolates, 79.5%, 38.5%, 17.9%, 10.3%, and 7.7% were, respectively, resistant, to nalidixic acid, ciprofloxacin, amoxicillin, erythromycin, and gentamicin. Among the 37 isolates of C. coli, 78.4%, 43.2%, 13.5%, 8.1%, and 0% were resistant, respectively, to the same antibiotics. In conclusion, we reported in this study the presence of high Campylobacter contamination of the studied chickens. Molecular identification of the bacteria was performed and determination of high resistance to antimicrobials of the fluoroquinolone family was revealed.

  4. Comparative genotyping of Campylobacter jejuni strains from patients with Guillain-Barre syndrome in Bangladesh.

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    Zhahirul Islam

    Full Text Available BACKGROUND: Campylobacter jejuni is a common cause of acute gastroenteritis and is associated with post-infectious neuropathies such as the Guillain-Barré syndrome (GBS and the Miller Fisher syndrome (MFS. We here present comparative genotyping of 49 C. jejuni strains from Bangladesh that were recovered from patients with enteritis or GBS. All strains were serotyped and analyzed by lipo-oligosaccharide (LOS genotyping, amplified fragment length polymorphism (AFLP analysis, multilocus sequence typing (MLST, and pulsed-field gel electrophoresis (PFGE. METHODOLOGY/PRINCIPAL FINDINGS: C. jejuni HS:23 was a predominant serotype among GBS patients (50%, and no specific serotype was significantly associated with GBS compared to enteritis. PCR screening showed that 38/49 (78% of strains could be assigned to LOS classes A, B, C, or E. The class A locus (4/7 vs 3/39; p<0.01 was significantly associated in the GBS-related strains as compared to enteritis strains. All GBS/oculomotor related strains contained the class B locus; which was also detected in 46% of control strains. Overlapping clonal groups were defined by MLST, AFLP and PFGE for strains from patients with gastroenteritis and GBS. MLST defined 22 sequence types (STs and 7 clonal complexes including 7 STs not previously identified (ST-3742, ST-3741, ST-3743, ST-3748, ST-3968, ST-3969 and ST-3970. C. jejuni HS:23 strains from patients with GBS or enteritis were clonal and all strains belonged to ST-403 complex. Concordance between LOS class B and ST-403 complex was revealed. AFLP defined 25 different types at 90% similarity. The predominant AFLP type AF-20 coincided with the C. jejuni HS:23 and ST-403 complex. CONCLUSION/SIGNIFICANCE: LOS genotyping, MLST, AFLP and PFGE helped to identify the HS:23 strains from GBS or enteritis patients as clonal. Overall, genotypes exclusive for enteritis or for GBS-related strains were not obtained although LOS class A was significantly associated with GBS

  5. ISOLASI Campylobacter DARI KARKAS AYAM MENGGUNAKAN METODE KONVENSIONAL DAN POLYMERASE CHAIN REACTIONS [Isolation of Campylobacter from Poultry Carcasses using Conventional and Polymerase Chain Reaction Methods

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    Surachmi Setiyaningsih2

    2013-06-01

    Full Text Available Campylobacter jejuni and Campylobacter coli are two spesies of Campylobacter sp. frequently found as pathogenic bacteria causing human gastrointestinal infections. Contaminated chicken carcasses have been reported as the source of human campylobacteriosis. In this study, Campylobacter were isolated from chicken carcasses sold in traditional markets and supermarkets. In traditional markets, chicken carcasses are sold without proper packaging or in an open space and stored at room temperature (25-30°C for prolonged period allowing pathogenic bacteria to grow. While at supermarkets, chicken carcasses are openly displayed or enclosed in plastic wrappings and stored in a refrigerator (4-8°C. A total of 298 samples of chicken carcasses from traditional markets and supermarkets in the area of DKI Jakarta, West Java (Bogor and Sukabumi and Central Java (Kudus and Demak were collected. Isolation and identification using conventional and Polymerase Chain Reactions (PCR methods were done to determine the prevalence of C. jejuni and C. coli contamination in poultry. The results showed that chicken carcasses sold in the sampling area, both traditional markets and supermarkets, were contaminated with C. jejuni and C. coli. The contamination rate of Campylobacter sp. in chicken carcasses sold in supermarkets, were 14.1% by conventional methods and 29.5% by PCR. This was higher than those in traditional markets, i.e. 5.7 and 12.1%, respectively. It is also confirmed that the prevalence for contamination of C. jejuni was higher than C. coli in 298 samples, i.e. 16.1% and 3.7% by conventional method and 23.5% and 18.1% by PCR method respectively.

  6. Occurrence, species distribution and antimicrobial resistance of thermophilic Campylobacter isolates from farm and laboratory animals in Morogoro, Tanzania

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    Erick V. G. Komba

    2014-08-01

    Full Text Available Aim: To determine the carriage and antimicrobial resistance of Thermophilic Campylobacter species in the gastrointestinal tracts of farm and laboratory animals in Morogoro, Tanzania Materials and Methods: Faecal samples were collected from farm (n=244 and laboratory (n=466 animals and were subjected to the Cape Town protocol for isolation of Campylobacter. Isolates were preliminarily identified based on potassium hydroxide string and hippurate hydrolysis tests. Polymerase chain reaction (PCR was employed for confirmation of isolates. Antimicrobial resistance testing was done using disc diffusion method. Results: Of the laboratory animals 26.7% of guinea pigs (n=30 and 1.2% of rats (n=242 were colonized with Campylobacter. Four isolates from guinea pigs were Campylobacter jejuni and the other four were C. coli. From rats, two isolates were C. jejuni and one was C. coli. In farm animals thermophilic Campylobacter were detected from 31.6% of sheep (n=57 and 60% horses (n=5. Of the isolates 12 (57% were C. jejuni (10 from sheep and 2 from horses and the remaining were C. coli (8 from sheep and 1 from a horse. The isolates were frequently resistant to erythromycin, norfloxacin, colistin sulphate and nalidixic acid; whereas low levels of resistance were observed for ciprofloxacin and gentamicin. Conclusion: Our study reveals carriage of antimicrobial resistant thermophilic Campylobacter in the intestines of the study animals. This highlights possibilities in involvement of these animals in the epidemiology of Campylobacter infections. Thus, there is a need to consider these animal species when planning control measures for this zoonotic bacterium.

  7. Antibacterial activity of three medicinal Thai plants against Campylobacter jejuni and other foodborne pathogens.

    Science.gov (United States)

    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

  8. Lectin typing of Campylobacter jejuni using a novel quartz crystal microbalance technique

    Energy Technology Data Exchange (ETDEWEB)

    Yakovleva, Maria E., E-mail: maria.yakovleva@gmail.com [Department of Infectious Diseases and Medical Microbiology, Lund University, 223 62 Lund (Sweden); Moran, Anthony P. [Department of Microbiology, School of Natural Sciences, National University of Ireland, Galway (Ireland); Safina, Gulnara R. [Department of Analytical and Marine Chemistry, University of Gothenburg, 412 96 Gothenburg (Sweden); Wadstroem, Torkel [Department of Infectious Diseases and Medical Microbiology, Lund University, 223 62 Lund (Sweden); Danielsson, Bengt [Acromed Invest AB, Magistratsvaegen 10, 226 43 Lund (Sweden)

    2011-05-23

    Seven Campylobacter jejuni strains were characterised by a lectin typing assay. The typing system was based on a quartz crystal microbalance technique (QCM) with four commercially available lectins (wheat germ agglutinin, Maackia amurensis lectin, Lens culinaris agglutinin, and Concanavalin A), which were chosen for their differing carbohydrate specificities. Initially, the gold surfaces of the quartz crystals were modified with 11-mercaptoundecanoic acid followed by lectin immobilisation using a conventional amine-coupling technique. Bacterial cells were applied for lectin typing without preliminary treatment, and resonant frequency and dissipation responses were recorded. The adhesion of microorganisms on lectin surfaces was confirmed by atomic force microscopy. Scanning was performed in the tapping mode and the presence of bacteria on lectin-coated surfaces was successfully demonstrated. A significant difference in the dissipation response was observed for different C. jejuni strains which made it possible to use this parameter for discriminating between bacterial strains. In summary, the QCM technique proved a powerful tool for the recognition and discrimination of C. jejuni strains. The approach may also prove applicable to strain discrimination of other bacterial species, particularly pathogens.

  9. Genome and Proteome of Campylobacter jejuni Bacteriophage NCTC 12673▿†

    Science.gov (United States)

    Kropinski, Andrew M.; Arutyunov, Denis; Foss, Mary; Cunningham, Anna; Ding, Wen; Singh, Amit; Pavlov, Andrey R.; Henry, Matthew; Evoy, Stephane; Kelly, John; Szymanski, Christine M.

    2011-01-01

    Campylobacter jejuni continues to be the leading cause of bacterial food-borne illness worldwide, so improvements to current methods used for bacterial detection and disease prevention are needed. We describe here the genome and proteome of C. jejuni bacteriophage NCTC 12673 and the exploitation of its receptor-binding protein for specific bacterial detection. Remarkably, the 135-kb Myoviridae genome of NCTC 12673 differs greatly from any other proteobacterial phage genome described (including C. jejuni phages CP220 and CPt10) and instead shows closest homology to the cyanobacterial T4-related myophages. The phage genome contains 172 putative open reading frames, including 12 homing endonucleases, no visible means of packaging, and a putative trans-splicing intein. The phage DNA appears to be strongly associated with a protein that interfered with PCR amplification and estimation of the phage genome mass by pulsed-field gel electrophoresis. Identification and analyses of the receptor-binding protein (Gp48) revealed features common to the Salmonella enterica P22 phage tailspike protein, including the ability to specifically recognize a host organism. Bacteriophage receptor-binding proteins may offer promising alternatives for use in pathogen detection platforms. PMID:21965409

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

    Science.gov (United States)

    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.

  11. Antibacterial effect of trans-cinnamaldehyde, eugenol, carvacrol, and thymol on Salmonella Enteritidis and Campylobacter jejuni in chicken cecal contents in vitro

    Science.gov (United States)

    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...

  12. Acute Pancreatitis in Association with Campylobacter jejuni- Associated Diarrhea in a 15-Year-Old with CFTR Mutations: Is There a Link?

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    Leena Kandula

    2006-09-01

    Full Text Available Context: Acute pancreatitis has occasionally been reported in association with Campylobacter jejuni infection in humans. However, the mechanism linking Campylobacter jejuni infection and pancreatitis isunclear. Acute pancreatitis in association with an infectious illness may be related to underlying genetic mutations. For instance, studies show that mutations in the cystic fibrosis transmembrane conductance regulator gene increase the susceptibility for acute and chronic pancreatitis. Case report :We describe a patient with Campylobacter jejuni infection who developed acute pancreatitis in the setting of an underlying cystic fibrosis transmembrane conductance regulator gene mutation. Discussion :In this patient with an underlying mutation in the CFTR gene, we propose that the interaction between the mutant gene and an environmental factor, Campylobacter jejuni infection, resulted in pancreatitis.

  13. Evaluating 3 gas-delivery systems for culturing Campylobacter jejuni in a microaerophilic environment.

    Science.gov (United States)

    Haines, M D; Eberle, K N; McDaniel, C D; Kiess, A S

    2011-10-01

    Campylobacter spp. require a microaerophilic environment (80% N(2), 10% CO(2), 5% H(2), and 5% O(2)) for growth. Since the late 1800s, several systems for creating and maintaining specific microbial atmospheres have been developed and applied. The objective of this study was to evaluate Campylobacter jejuni growth by means of 3 commonly used gas-delivery systems for generating a microaerophilic environment: automated, gas-generating sachet, and plastic storage bag. Pure culture C. jejuni cells were suspended in Brucella broth and spread onto campy cefex agar plates. For the automated gas-delivery system, plates were positioned in a Mart anaerobic jar and flushed with a microaerophilic gas mixture using an Anoxomat Mart II system (Mart Microbiology B. V., Netherlands). For the sachet samples, plates were placed in a Mart anaerobic jar and 3 Gaspak EZ campy sachets (Becton Dickinson and Company, Franklin Lakes, NJ) were activated to induce a microaerophilic gas environment. The plates placed in plastic storage bags were flushed with a microaerophilic gas mixture from a premixed tank. For all 3 systems, plates were placed in a low-temperature incubator at 42°C for 24 h. After 24 h, plates were removed from the incubator and colonies were counted. The entire experiment was repeated 5 times. Results indicated no significant difference in colony counts among the gas-delivery systems tested, but colonies grown under the sachet-generated environment were smaller than colonies in the other 2 methods. Smaller colonies could have resulted from the type of media used or the length of time the plates were incubated. In conclusion, all 3 gas-delivery methods were able to produce similar Campylobacter growth results. Initial and long-term costs of equipment, as well as laboratory space availability, may be influential when choosing a gas-delivery method for generating a microaerophilic environment. PMID:21934023

  14. Antibodies to Glycoproteins Shared by Human Peripheral Nerve and Campylobacter jejuni in Patients with Multifocal Motor Neuropathy

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    Ljubica Suturkova

    2013-01-01

    Full Text Available We have tested serum samples from 24 patients with multifocal motor neuropathy (MMN for reactivity to ganglioside GM1 and to Gal(β1–3GalNAc-bearing glycoproteins isolated from human peripheral nerve and from Campylobacter jejuni (Cj serotype O:19. IgM anti-GM1 antibodies were detected by ELISA in 11 patients (45.8% with MMN and in only one subject (4% from the control group. Western blots showed positive reactivity of sera from 6 patients (25% with MMN to several Gal(β1–3GalNAc-bearing glycoproteins from human peripheral nerve and from Cj O:19 isolates. Sera from three patients (12.5% with MMN showed positively reactive bands with similar electrophoretic mobility in all isolates (60–62 kDa, 48–51 kDa, 42 kDa, and 38 kDa. All six patients showed positive reactivity to 48–52 kDa protein isolated from human peripheral nerve. Increased titer of IgG antibodies to 60–62 kDa protein isolated from Cj O:19 associated with Guillain-Barré syndrome was detected in three patients, and their serum showed also IgG positive reactivity to peripheral nerve antigen with the same electrophoretic mobility. One of these patients had a previous history of Cj infection which suggests the possibility that Cj may be also involved in the pathogenesis of MMN.

  15. Campylobacter jejuni CsrA complements an Escherichia coli csrA mutation for the regulation of biofilm formation, motility and cellular morphology but not glycogen accumulation

    OpenAIRE

    Fields Joshua A; Thompson Stuart A

    2012-01-01

    Abstract Background Although Campylobacter jejuni is consistently ranked as one of the leading causes of bacterial diarrhea worldwide, the mechanisms by which C. jejuni causes disease and how they are regulated have yet to be clearly defined. The global regulator, CsrA, has been well characterized in several bacterial genera and is known to regulate a number of independent pathways via a post transcriptional mechanism, but remains relatively uncharacterized in the genus Campylobacter. Previou...

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

    NARCIS (Netherlands)

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

    2016-01-01

    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 contam

  17. Campylobacter jejuni and Campylobacter coli in wild birds on Danish livestock farms

    DEFF Research Database (Denmark)

    Hald, Birthe; Skov, Marianne Nielsine; Nielsen, Eva Møller;

    2016-01-01

    order to study the factors influencing the prevalence in wild birds according to their ecological guild. In total, 1607 individual wild bird cloacal swab samples and 386 livestock manure samples were cultured for Campylobacter spp. according to the Nordic Committee on Food Analysis method NMKL 119...

  18. Does age acquired immunity confer selective protection to common serotypes of Campylobacter jejuni?

    Directory of Open Access Journals (Sweden)

    Ogden Iain D

    2005-08-01

    Full Text Available Abstract Background Campylobacter infection is a major cause of bacterial gastrointestinal disease. Exposure to Campylobacter is known to produce an immune response in humans that can prevent future symptomatic infections. Further, studies of the general population have shown that seroprevalence to Campylobacter increases with age. Methods A large collection of serotyped Campylobacter isolates, obtained from human clinical faecal samples, were analysed by comparing the ratio of uncommon to common serotypes by different age groups, using χ2 tests. Results We have identified that older age groups, as well as having generally lower incidence, are significantly less likely to be infected by the more common serotypes. Conclusion These results are indicative of acquired immunity, however, further studies are needed to rule out the confounding effects of the variations in exposure pathways experienced by different age groups.

  19. The ability of flagellum-specific Proteus vulgaris bacteriophage PV22 to interact with Campylobacter jejuni flagella in culture

    Directory of Open Access Journals (Sweden)

    Stern NJ

    2006-06-01

    Full Text Available Abstract Background There has been a recent resurgent interest in bacteriophage biology. Research was initiated to examine Campylobacter jejuni-specific bacteriophage in the Russian Federation to develop alternative control measures for this pathogen. Results A C. jejuni flagellum-specific phage PV22 from Proteus vulgaris was identified in sewage drainage. This phage interacted with C. jejuni by attachment to flagella followed by translocation of the phage to the polar region of the bacterium up to the point of DNA injection. Electron microscopic examination revealed adsorption of PV22 on C. jejuni flagella after a five minute incubation of the phage and bacteria. A different phenomenon was observed after incubating the mix under the same conditions, but for twenty minutes or longer. Phage accumulated primarily on the surface of cells at sites where flagella originated. Interestingly, PV22 did not inject DNA into C. jejuni and PV22 did not produce lytic plaques on medium containing C. jejuni cells. The constant of velocity for PV22 adsorption on cells was 7 × 10-9 ml/min. Conclusion It was demonstrated that a bacteriophage that productively infects P. vulgaris was able to bind C. jejuni and by a spot test that the growth of C. jejuni was reduced relative to control bacteria in the region of phage application. There may be two interesting applications of this effect. First, it may be possible to test phage PV22 as an antimicrobial agent to decrease C. jejuni colonization of the chicken intestine. Second, the phage could potentially be utilized for investigating biogenesis of C. jejuni flagella.

  20. Can Campylobacter jejuni play a role in development of celiac disease? A hypothesis

    Institute of Scientific and Technical Information of China (English)

    Behnam Sabayan; Farzaneh Foroughinia; Mohammad Hadi Imanieh

    2007-01-01

    Celiac disease (CD) is an entropathy with malabsortive condition in which an allergic reaction to the cereal grain-protein (gluten) causes small intestine mucosal injury. CD is a multifactorial disorder in which both genetic and environmental factors contribute to the disease development. Mechanisms have been described to explain the pathology of CD. T cells specific for multiple gluten peptides are found in virtually all patients. Generation of such a broad T cell response may be a prerequisite for disease development. CD is associated with multiple extraintestinal presentations,including neurological deficits. Recent studies have shown a significant correlation between anti-ganglioside antibodies and neurological disorders in patients with underlying CD. Gangliosides are glycosphingolipids which are abundant in nervous system and in other tissues including gastrointestinal tract. It is not known what triggers the release of anti-ganglioside antibodies in people with gluten sensitivity. But, the mechanism is likely to involve the intestinal immune system response to ingested gliadin, a component of wheat gluten. Studies showed that mechanisms different from gluten exposure may be implicated in antibody formation, and other environmental factors may also exist. In addition, considering the fact that genetic predisposition dysregulating mucosal immune responses in the presence of certain environmental triggers like gastrointestinal infections may be strong etiological factors for developing chronic intestinal inflammation including CD, the hypothesis raised in our mind that antiganglioside antibody formation in CD may play a role not only in development of neurological complications in celiac patients, but also in development of CD itself. As presence of Campylobacter jejuni in other diseases with antigangliosides antibody formation has been established, we propose the possible role of Campylobacter jejuni in development of CD in association with other genetic and

  1. Genotypes, Antibiotic Resistance, and ST-8 Genetic Clone in Campylobacter Isolates from Sheep and Goats in Grenada

    Directory of Open Access Journals (Sweden)

    Diana M. Stone

    2014-01-01

    Full Text Available Rectal swabs from 155 sheep and 252 goats from Grenada were evaluated to determine the prevalence of Campylobacter spp., antibiotic resistance, and multilocus sequence types. Fifteen Campylobacter isolates were obtained (14 C. jejuni and 1 C. coli. The prevalence (3.7% did not differ significantly between sheep (4.5% and goats (3.2%. Among the seven antimicrobials tested, resistance was only detected for tetracycline (30.8% and metronidazole (38.5%. Campylobacter isolates showed no significant difference between sheep and goats for type of antimicrobial resistance or percent of resistant isolates. Twelve of the isolates were successfully genotyped consisting of four recognized clonal complexes and three novel sequence types. Importantly, one isolate from one goat was identified as the C. jejuni sequence type-8, a zoonotic and tetracycline-resistant clone reported to be a highly virulent clone associated with ovine abortion in the USA. Although most samples were from comingled sheep and goat production units, there were no shared sequence types between these two host species. None of the sequence types identified in this study have previously been reported in poultry in Grenada, suggesting sheep- and goat-specific Campylobacter clones in Grenada. This is the first report of genotyping of Campylobacter isolates from sheep and goats in the Eastern Caribbean.

  2. Crystal structure of the Campylobacter jejuni Cj0090 protein reveals a novel variant of the immunoglobulin fold among bacterial lipoproteins.

    Science.gov (United States)

    Paek, Seonghee; Kawai, Fumihiro; Choi, Kyoung-Jae; Yeo, Hye-Jeong

    2012-12-01

    Bacterial lipoproteins play an important role in bacterial pathogenesis and physiology. The genome of Campylobacter jejuni, a major foodborn pathogen, is predicted to contain over 20 lipoproteins. However, the functions of the majority of C. jejuni lipoproteins remain unknown. The Cj0090 protein is encoded by a lipoprotein operon composed of cj0089, cj0090, and cj0091. Here, we report the crystal structure of Cj0090 at 1.9 Å resolution, revealing a novel variant of the immunoglobulin fold with β-sandwich architecture. The structure suggests that Cj0090 may be involved in protein-protein interactions, consistent with a possible role for bacterial lipoproteins. PMID:22987763

  3. Comparative Proteomics and Glycoproteomics Reveal Increased N-Linked Glycosylation and Relaxed Sequon Specificity in Campylobacter jejuni NCTC11168 O

    DEFF Research Database (Denmark)

    Scott, Nichollas E.; Marzook, N. Bishara; Cain, Joel A.;

    2014-01-01

    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 Pgl......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...

  4. Interaction between Food-borne Pathogens (Campylobacter jejuni, Salmonella Typhimurium and Listeria monocytogenes) and a Common Soil Flagellate (Cercomonas sp.)

    DEFF Research Database (Denmark)

    Bui, Thanh Xuan; Wolff, Anders; Madsen, Mogens;

    2012-01-01

    Free-living protozoa may harbor, protect, and disperse bacteria, including those ingested and passed in viable form in feces. The flagellates are very important predators on bacteria in soil, but their role in the survival of food-borne pathogens associated with fruits and vegetables is not well...... understood. In this study, we investigated the interactions between a common soil flagellate, Cercomonas sp., and three different bacterial pathogens (Campylobacter jejuni, Salmonella Typhimurium, and Listeria monocytogenes). Rapid growth of flagellates was observed in co-culture with C. jejuni and S...

  5. Short-chain fatty acids and L-lactate as feed additives to control Campylobacter jejuni infections in broilers.

    Science.gov (United States)

    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

  6. Campylobacter jejuni induces diverse kinetics and profiles of cytokine genes in INT-407 cells

    International Nuclear Information System (INIS)

    Objective was to examine the kinetic ability of embryonic human epithelial INT-407 cells to express messenger ribonucleic acid (mRNA) for various cytokines and chemokines in response to Campylobacter jejuni (C. jejuni) stimulation. In an experimental single-blind study, cultured embryonic human epithelial INT-407 cells were treated with different concentrations of viable C. jejuni, its sonicated and filtered supernatant. A modified non-radioactive in situ hybridization using probe cocktails was used to measure mRNA levels for the pro-inflammatory cytokines interleukin (IL)-1beta, IL-6, interferon-gamma, tumor necrosis factor (TNF)-alpha, transforming growth factor (TGF)-beta1 and IL-8 and the anti-inflammatory cytokines, IL-4 and IL-10. The study was carried out from September 2005 to March 2007 at the Department of Microbiology, Immunology and Infectious Diseases, College of Medicine and Medical Sciences, Arabian Gulf University, Bahrain. Viable C. jejuni sonicated bacteria and filtered supernatant induced high mRNA expression for the pro-inflammatory cytokines IL-1 beta, IL-6, IFN-gama, TNF-alpha, TGF-beta and IL-8 which peaked at the 12 hours post stimulation. Anti-inflammatory cytokine IL-4 and IL-10 mNRA expression were induced maximally at 3 hours post stimulation mainly by sonicated bacteria and filtrated supernatant, however, not with living bacteria and filtrated supernatant, however, not with living bacteria. Untreated embryonic human epithelial INT-407 cells expressed low amount of mNRA for the various cytokines and chemokines at all time points. For each cytokine, 4 samples were used per time hour. This study demonstrated that embryonic human epithelial INT-407 cells in response to viable C. jejuni or its cytotxins can alter cytokine and chemokine mNRA expression patterns and kinetics suggesting a potential role for these mediators in the immunopathogenesis of the infection caused by this pathogen, which might be relevant for future immunotherapeutic

  7. 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.

    Science.gov (United States)

    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...

  8. In vivo broiler experiments to assess anti-Campylobacter jejuni activity of a live Enterococcus faecalis strain.

    Science.gov (United States)

    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

  9. Preliminary structural studies of the transcriptional regulator CmeR from Campylobacter jejuni

    Energy Technology Data Exchange (ETDEWEB)

    Su, Chih-Chia [Department of Biochemistry, Biophysics and Molecular Biology, Iowa State University, Ames, IA 50011 (United States); Shi, Feng [Department of Veterinary Microbiology, College of Veterinary Medicine, Iowa State University, Ames, IA 50011 (United States); Gu, Ruoyu; Li, Ming [Department of Physics and Astronomy, Iowa State University, Ames, IA 50011 (United States); McDermott, Gerry [Department of Anatomy, School of Medicine, University of California, San Francisco, CA 94143 (United States); Yu, Edward W., E-mail: ewyu@iastate.edu [Department of Biochemistry, Biophysics and Molecular Biology, Iowa State University, Ames, IA 50011 (United States); Department of Physics and Astronomy, Iowa State University, Ames, IA 50011 (United States); Zhang, Qijing [Department of Veterinary Microbiology, College of Veterinary Medicine, Iowa State University, Ames, IA 50011 (United States); Department of Biochemistry, Biophysics and Molecular Biology, Iowa State University, Ames, IA 50011 (United States)

    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.

  10. Investigations into the antibacterial activities of phytotherapeutics against Helicobacter pylori and Campylobacter jejuni.

    Science.gov (United States)

    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

  11. Specificity of Campylobacter jejuni Adhesin PEB3 for Phosphates and Structural Differences among Its Ligand Complexes

    Energy Technology Data Exchange (ETDEWEB)

    Min, Tongpil; Vedadi, Masoud; Watson, David C.; Wasney, Gregory A.; Munger, Christine; Cygler, Miroslaw; Matte, Allan; Young, N. Martin; (NRCC); (McGill); (Toronto)

    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.

  12. Campylobacter jejuni pdxA affects flagellum-mediated motility to alter host colonization.

    Directory of Open Access Journals (Sweden)

    Hiroshi Asakura

    Full Text Available 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.

  13. The effect of probiotics on broiler growth and intestinal morphology when used to prevent Campylobacter jejuni colonization

    Directory of Open Access Journals (Sweden)

    Lavinia Ştef

    2015-05-01

    Full Text Available The aim of this work was to establish the effect of probiotic microorganisms on growth performance and intestinal changes caused by Campylobacter jejuni colonization.In this respect, we used four probiotic microorganisms, namely: Lactobacillus paracasei JR, L. rhamnosus 15b, Y L. lactis and L. lactis FOA.The administration of probiotic microorganisms in different combinations and in different periods of growth does not significantly influence the bioproductive indices of broilers,that is,the total gain, feed intake and FCR (p>0.05. After studying the intestinal mucosa, it was concluded that the four microorganisms administered in broilers’s food determineschanges in the mucosa, inhibiting the development of Campylobacter jejuni,by the presence of smaller caliciform cells and the presence ofreduced leukocyte infiltration in the chorion of the mucosal.

  14. [Use of AFLP analysis for the molecular biological fine typing of Campylobacter jejuni strains from turkey flocks].

    Science.gov (United States)

    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

  15. Membrane integrity of Campylobacter jejuni subjected to high pressure is pH-dependent

    Science.gov (United States)

    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.

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

    Directory of Open Access Journals (Sweden)

    Euna eOh

    2015-04-01

    Full Text Available 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 (ROS 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 (VBNC 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.

  17. Complete Genome Sequence of Campylobacter iguaniorum Strain 1485ET, Isolated from a Bearded Dragon (Pogona vitticeps)

    OpenAIRE

    Gilbert, Maarten J; Miller, William G; Yee, Emma; Kik, Marja; Wagenaar, Jaap A.; Duim, Birgitta

    2014-01-01

    Campylobacter iguaniorum has been isolated from reptiles. This Campylobacter species is genetically related to Campylobacter fetus and Campylobacter hyointestinalis. Here we present the first whole-genome sequence for this species.

  18. Complete Genome Sequence of Campylobacter iguaniorum Strain 1485ET, Isolated from a Bearded Dragon (Pogona vitticeps).

    Science.gov (United States)

    Gilbert, Maarten J; Miller, William G; Yee, Emma; Kik, Marja; Wagenaar, Jaap A; Duim, Birgitta

    2014-01-01

    Campylobacter iguaniorum has been isolated from reptiles. This Campylobacter species is genetically related to Campylobacter fetus and Campylobacter hyointestinalis. Here we present the first whole-genome sequence for this species. PMID:25146144

  19. Divergent distribution of the sensor kinase CosS in non-thermotolerant campylobacter species and its functional incompatibility with the response regulator CosR of Campylobacter jejuni.

    Directory of Open Access Journals (Sweden)

    Sunyoung Hwang

    Full Text Available 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

  20. Comparative Study of Campylobacter spp. Isolated from Children With Gastroenteritis in Bahonar Hospital, Karaj, Using PCR and RFLP

    Directory of Open Access Journals (Sweden)

    Abdi

    2016-05-01

    Full Text Available Background Campylobacter species are responsible for the majority of cases of food-borne gastroenteritis. The sources of the disease outbreaks are often contaminated water or milk, and consumption of undercooked poultry product is the main cause of sporadic campylobacteriosis cases. Objectives The aims of this study were to determine the prevalence of Campylobacter gastroenteritis in children and to differentiate the interfering species using polymerase chain reaction (PCR and restriction fragment length polymorphism (RFLP methods at the Bahonar hospital in Karaj, Iran. Patients and Methods A total of 150 stool samples were collected from children under 10 years old during the summer of 2014. PCR was performed using genus- and species-specific primers and RFLP was done using AluI and TasI enzymes. Results The results showed the amplification of 400 and 491 bp segments and Campylobacter contamination in 30 (20% samples; 5 out of 30 Campylobacter positive samples (16.66% were identified as C. jejuni, 20 (66.66% as C. coli, 3 (10% as C. jejuni and C. coli (mixed infection, and 2 (6.66% were identified as non-jejuni, non-coli Campylobacter using the PCR method. Following the evaluation of RFLP results, 7 positive samples (23.33% showed the electrophoretic pattern of C. jejuni, 21 (70% showed the electrophoretic pattern of C. coli, and 2 (6.6% showed both of the patterns and mixed contamination with jejuni and coli species. The results of digestion with TasI did not show any C. lari or C. upsaliensis patterns. Conclusions The results of this study showed high percentage of Campylobacter contamination in the tested stool samples. The other surprising finding was the high rate of Campylobacter coli positive samples; the difference between the results of PCR using species-specific primers (hipo and asp and the RFLP method (electrophoretic patterns in some of the positive samples confirms the hypothesis of variations in nucleotide sequences of the hipo

  1. Peripheral CD4+ T cell cytokine responses following human challenge and re-challenge with Campylobacter jejuni.

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    Kelly A Fimlaid

    Full Text Available Campylobacter jejuni is a leading cause of human gastroenteritis worldwide; however, our understanding of the human immune response to C. jejuni infection is limited. A previous human challenge model has shown that C. jejuni elicits IFNγ production by peripheral blood mononuclear cells, a response associated with protection from clinical disease following re-infection. In this study, we investigate T lymphocyte profiles associated with campylobacteriosis using specimens from a new human challenge model in which C. jejuni-naïve subjects were challenged and re-challenged with C. jejuni CG8421. Multiparameter flow cytometry was used to investigate T lymphocytes as a source of cytokines, including IFNγ, and to identify cytokine patterns associated with either campylobacteriosis or protection from disease. Unexpectedly, all but one subject evaluated re-experienced campylobacteriosis after re-challenge. We show that CD4+ T cells make IFNγ and other pro-inflammatory cytokines in response to infection; however, multifunctional cytokine response patterns were not found. Cytokine production from peripheral CD4+ T cells was not enhanced following re-challenge, which may suggest deletion or tolerance. Evaluation of alternative paradigms or models is needed to better understand the immune components of protection from campylobacteriosis.

  2. Campylobacter infection

    Science.gov (United States)

    ... 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 ...

  3. Non-selective regulation of peroxide and superoxide resistance genes by PerR in Campylobacter jejuni

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    Jong-Chul eKim

    2015-02-01

    Full Text Available Campylobacter jejuni is an important foodborne pathogen. The molecular mechanisms for the regulation of oxidative stress resistance have not yet been understood fully in this bacterium. In this study, we investigated how PerR (peroxide stress regulator modulates the transcriptional regulation of both peroxide and superoxide resistance genes in C. jejuni, particularly under oxidative stress conditions. The transcriptional levels of ahpC, katA, and sodB were substantially increased by aeration and oxidant exposure. Interestingly, a perR mutation completely abrogated the transcriptional response of ahpC, katA and sodB to oxidants. Furthermore, we demonstrated that perR transcription was reduced by aeration and oxidant exposure. In contrast to the unique role of PerR homologs in peroxide stress regulation in other bacteria, interestingly, C. jejuni PerR directly regulates the transcription of sodB, the most important gene in superoxide defense, as evidenced by the alteration of sodB transcription by the perR mutation and direct binding of rPerR to the sodB promoter. In addition, we also observed notable morphological changes in C. jejuni from spiral rods to coccoid morphology under aerobic conditions. Based on the intracellular ATP levels, C. jejuni entered a viable-but-non-culturable state under aerobic conditions. These findings clearly demonstrate that C. jejuni possesses a unique regulatory mechanism of oxidative stress defense that does not specifically distinguish between peroxide and superoxide defense, and PerR plays a pivotal role in this non-selective regulation of oxidative stress resistance in C. jejuni.

  4. 广西南宁市腹泻人群空肠弯曲菌流行状况调查%Epidemic status investigation of campylobacter jejuni in the patients with diarrhea living in Nanning, Guangxi

    Institute of Scientific and Technical Information of China (English)

    2013-01-01

    Objective To determine the infectious status of campylobacter jejuni in the patients with diarrhea who lived in Nanning. Methods Epidemiologic investigation was conducted on the diarrheal patients living in Nanning, Their stool samples were collected and cultured for bacterial pathogens and were analyzed by means of Polymerase chain reaction (PCR) and Pulsed field gel electrophoresis (PFGE). Results Campylobacter jejuni was isolated from 3 of 775 stool samples with a positive rate of 0.39%. All positive isolates for biochemical tests were also positive for PCR reconfirmation. The PFGE patterns of 3 Campylobacter jejuni isolates belonged to 3 different clusters. Conclusion The cases infected with campylobacter jejuni have been identified in Nanning.The results of this study have provided scientific basis for appropriate evaluation of epidemic status of campylobacter jejuni in diarrheal patients in Nanning and for developing control measures.%  目的了解南宁市腹泻人群中空肠弯曲菌的感染流行状况。方法在南宁市对腹泻病例进行流行病学调查并采集粪便标本,进行实验室空弯菌的分离培养、PCR鉴定以及脉冲场凝胶电泳分型(PFGE)分析。结果在775份粪便样品中,3份空肠弯曲菌阳性,平均阳性率0.39%。所有生化结果阳性的菌株其PCR复核鉴定结果均为阳性。3株空肠弯曲菌经Cluster分析后得到3个PFGE带型。结论南宁市存在空肠弯曲菌感染病例,本研究结果为正确评价南宁市腹泻人群弯曲菌的流行状况和制定防控措施提供了科学依据。

  5. Rapid identification of novel immunodominant proteins and characterization of a specific linear epitope of Campylobacter jejuni.

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    Sebastian Hoppe

    Full Text Available 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

  6. Rapid Identification of Novel Immunodominant Proteins and Characterization of a Specific Linear Epitope of Campylobacter jejuni

    Science.gov (United States)

    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

  7. Eukaryotic Cell Invasion does not correlate to flaA SVR Sequence Type based on a Library of Genetically Diverse Campylobacter jejuni Isolates Originally Recovered from A Variety of Sources in Iceland

    Science.gov (United States)

    Introduction: Campylobacter spp. are considered to be a leading bacterial etiologic agent of acute food-borne gastroenteritis among human populations. Epithelial cell invasion is hypothesized to be necessary for human infection and cell invasion assays have been utilized to demonstrate that distinc...

  8. The influence of feeding crimped kernel maize silage on growth performance and intestinal colonization with Campylobacter jejuni of broilers.

    Science.gov (United States)

    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.

  9. Point mutations in the major outer membrane protein drive hypervirulence of a rapidly expanding clone of Campylobacter jejuni.

    Science.gov (United States)

    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

  10. FROM GENE TO PROTEIN – CLONNING, EXPRESSION AND PUFICATION OF A P450 CYTOCHROM FROM Campylobacter jejuni

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    N. CORCIONIVOSCHI

    2013-07-01

    Full Text Available Recently, the complete genome sequence of Campylobacter jejuni NCTC 11168 was published revealing the presence of only one open reading frame (Cj1411c encoding for a cytochrome P450, in contrast to 20 found in M. tuberculosis. The gene Cj1411c encodes for a soluble 52.6 kDa protein with a predicted isoelectric point of 9.3. The P450 gene is part of reading frame which hosts genes involved in the synthesis of cell surface components (capsula. Campylobacter capsule are important in adherence, invasion and colonisation of host cells and for maintenance of cell surface charge and serum resistance. These capsule are thought to cause autoimmunity leading to Guillan-Barre and Miller-Fischer syndromes. The structure of the lipoolygosaccharides and capsule polysaccharide was published last year revealing that the strain possessed a type II/III capsule locus found in other microorganisms such Nisseria meningitidis. This project focuses on the cloning and characterisation of the only P450 enzyme of the human pathogen Campylobacter jejuni NCTC 11168. We aim to understand the metabolic role of this P450 cytochrome in order to elucidate its possible use as a new target for drug design. To achieve this aim we have cloned, expressed and purify the product of P450 coding gene.

  11. FROM GENE TO PROTEIN – CLONNING, EXPRESSION AND PUFICATION OF A P450 CYTOCHROM FROM CAMPYLOBACTER JEJUNI

    Directory of Open Access Journals (Sweden)

    N. CORCIONIVOSCHI

    2009-05-01

    Full Text Available Recently, the complete genome sequence of Campylobacter jejuni NCTC 11168 was published revealingthe presence of only one open reading frame (Cj1411c encoding for a cytochrome P450, in contrast to 20found in M. tuberculosis. The gene Cj1411c encodes for a soluble 52.6 kDa protein with a predictedisoelectric point of 9.3. The P450 gene is part of reading frame which hosts genes involved in thesynthesis of cell surface components (capsula. Campylobacter capsule are important in adherence,invasion and colonisation of host cells and for maintenance of cell surface charge and serum resistance.These capsule are thought to cause autoimmunity leading to Guillan-Barre and Miller-Fischersyndromes. The structure of the lipoolygosaccharides and capsule polysaccharide was published last yearrevealing that the strain possessed a type II/III capsule locus found in other microorganisms suchNisseria meningitidis. This project focuses on the cloning and characterisation of the only P450 enzymeof the human pathogen Campylobacter jejuni NCTC 11168. We aim to understand the metabolic role ofthis P450 cytochrome in order to elucidate its possible use as a new target for drug design. To achievethis aim we have cloned, expressed and purify the product of P450 coding gene.

  12. Crystal Structure and Catalytic Mechanism of PglD from Campylobacter jejuni

    Energy Technology Data Exchange (ETDEWEB)

    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.

  13. High Throughput Method for Analysis of Repeat Number for 28 Phase Variable Loci of Campylobacter jejuni Strain NCTC11168

    Science.gov (United States)

    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

  14. Campylobacter jejuni CsrA complements an Escherichia coli csrA mutation for the regulation of biofilm formation, motility and cellular morphology but not glycogen accumulation

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    Fields Joshua A

    2012-10-01

    Full Text Available Abstract Background Although Campylobacter jejuni is consistently ranked as one of the leading causes of bacterial diarrhea worldwide, the mechanisms by which C. jejuni causes disease and how they are regulated have yet to be clearly defined. The global regulator, CsrA, has been well characterized in several bacterial genera and is known to regulate a number of independent pathways via a post transcriptional mechanism, but remains relatively uncharacterized in the genus Campylobacter. Previously, we reported data illustrating the requirement for CsrA in several virulence related phenotypes of C. jejuni strain 81–176, indicating that the Csr pathway is important for Campylobacter pathogenesis. Results We compared the Escherichia coli and C. jejuni orthologs of CsrA and characterized the ability of the C. jejuni CsrA protein to functionally complement an E. coli csrA mutant. Phylogenetic comparison of E. coli CsrA to orthologs from several pathogenic bacteria demonstrated variability in C. jejuni CsrA relative to the known RNA binding domains of E. coli CsrA and in several amino acids reported to be involved in E. coli CsrA-mediated gene regulation. When expressed in an E. coli csrA mutant, C. jejuni CsrA succeeded in recovering defects in motility, biofilm formation, and cellular morphology; however, it failed to return excess glycogen accumulation to wild type levels. Conclusions These findings suggest that C. jejuni CsrA is capable of efficiently binding some E. coli CsrA binding sites, but not others, and provide insight into the biochemistry of C. jejuni CsrA.

  15. Characterization of putative cholesterol recognition/interaction amino acid consensus-like motif of Campylobacter jejuni cytolethal distending toxin C.

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    Chih-Ho Lai

    Full Text Available Cytolethal distending toxin (CDT produced by Campylobacter jejuni comprises a heterotrimeric complex formed by CdtA, CdtB, and CdtC. Among these toxin subunits, CdtA and CdtC function as essential proteins that mediate toxin binding to cytoplasmic membranes followed by delivery of CdtB into the nucleus. The binding of CdtA/CdtC to the cell surface is mediated by cholesterol, a major component in lipid rafts. Although the putative cholesterol recognition/interaction amino acid consensus (CRAC domain of CDT has been reported from several bacterial pathogens, the protein regions contributing to CDT binding to cholesterol in C. jejuni remain unclear. Here, we selected a potential CRAC-like region present in the CdtC from C. jejuni for analysis. Molecular modeling showed that the predicted functional domain had the shape of a hydrophobic groove, facilitating cholesterol localization to this domain. Mutation of a tyrosine residue in the CRAC-like region decreased direct binding of CdtC to cholesterol rather than toxin intermolecular interactions and led to impaired CDT intoxication. These results provide a molecular link between C. jejuni CdtC and membrane-lipid rafts through the CRAC-like region, which contributes to toxin recognition and interaction with cholesterol.

  16. Influence of Asellus aquaticus on Escherichia coli, Klebsiella pneumoniae, Campylobacter jejuni and naturally occurring heterotrophic bacteria in drinking water

    DEFF Research Database (Denmark)

    Christensen, Sarah Christine; Nissen, Erling; Arvin, Erik;

    2012-01-01

    . aquaticus on microbial water quality in non-chlorinated drinking water in controlled laboratory experiments. Pure cultures of the indicator organisms Escherichia coli and Klebsiella pneumoniae and the pathogen Campylobacter jejuni as well as naturally occurring heterotrophic drinking water bacteria...... decreased over time, following a first order decay with half lives of 5.3, 18.4 and 1.3 days, respectively. A. aquaticus did not affect survival of indicators and pathogens substantially whereas HPC were influenced by presence of dead A. aquaticus. Growth rates increased with an average of 48% for bacteria...... grown on R-2A agar and an average of 83% for bacteria grown on yeast extract agar when dead A. aquaticus were present compared to no and living A. aquaticus present. A. aquaticus associated E. coli, K. pneumoniae and C. jejuni were measured (up to 25 per living and 500 per dead A. aquaticus) and so were...

  17. Campylobacter jejuni fatty acid synthase II: Structural and functional analysis of [beta]-hydroxyacyl-ACP dehydratase (FabZ)

    Energy Technology Data Exchange (ETDEWEB)

    Kirkpatrick, Andrew S.; Yokoyama, Takeshi; Choi, Kyoung-Jae; Yeo, Hye-Jeong; (Houston)

    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

  18. Campylobacter jejuni increases flagellar expression and adhesion of noninvasive Escherichia coli: effects on enterocytic Toll-like receptor 4 and CXCL-8 expression.

    Science.gov (United States)

    Reti, Kristen L; Tymensen, Lisa D; Davis, Shevaun P; Amrein, Matthias W; Buret, Andre G

    2015-12-01

    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

  19. The cinnamon-oil ingredient trans-cinnamaldehyde fails to target Campylobacter jejuni strain KC 40 in the broiler chicken cecum despite marked in vitro activity.

    Science.gov (United States)

    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

  20. Avian resistance to Campylobacter jejuni colonization is associated with an intestinal immunogene expression signature identified by mRNA sequencing.

    Directory of Open Access Journals (Sweden)

    Sarah Connell

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

  1. Characterization of Antimicrobial Resistance Patterns and Detection of Virulence Genes in Campylobacter Isolates in Italy

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    Elisabetta Di Giannatale

    2014-02-01

    Full Text Available Campylobacter has developed resistance to several antimicrobial agents over the years, including macrolides, quinolones and fluoroquinolones, becoming a significant public health hazard. A total of 145 strains derived from raw milk, chicken faeces, chicken carcasses, cattle faeces and human faeces collected from various Italian regions, were screened for antimicrobial susceptibility, molecular characterization (SmaI pulsed-field gel electrophoresis and detection of virulence genes (sequencing and DNA microarray analysis. The prevalence of C. jejuni and C. coli was 62.75% and 37.24% respectively. Antimicrobial susceptibility revealed a high level of resistance for ciprofloxacin (62.76%, tetracycline (55.86% and nalidixic acid (55.17%. Genotyping of Campylobacter isolates using PFGE revealed a total of 86 unique SmaI patterns. Virulence gene profiles were determined using a new microbial diagnostic microarray composed of 70-mer oligonucleotide probes targeting genes implicated in Campylobacter pathogenicity. Correspondence between PFGE and microarray clusters was observed. Comparisons of PFGE and virulence profiles reflected the high genetic diversity of the strains examined, leading us to speculate different degrees of pathogenicity inside Campylobacter populations.

  2. Source attribution of human Campylobacter isolates by MLST and fla-typing and association of genotypes with quinolone resistance.

    Science.gov (United States)

    Kittl, Sonja; Heckel, Gerald; Korczak, Bożena M; Kuhnert, Peter

    2013-01-01

    Campylobacteriosis is the most frequent zoonosis in developed countries and various domestic animals can function as reservoir for the main pathogens Campylobacter jejuni and Campylobacter coli. In the present study we compared population structures of 730 C. jejuni and C. coli from human cases, 610 chicken, 159 dog, 360 pig and 23 cattle isolates collected between 2001 and 2012 in Switzerland. All isolates had been typed with multi locus sequence typing (MLST) and flaB-typing and their genotypic resistance to quinolones was determined. We used complementary approaches by testing for differences between isolates from different hosts with the proportion similarity as well as the fixation index and by attributing the source of the human isolates with Bayesian assignment using the software STRUCTURE. Analyses were done with MLST and flaB data in parallel and both typing methods were tested for associations of genotypes with quinolone resistance. Results obtained with MLST and flaB data corresponded remarkably well, both indicating chickens as the main source for human infection for both Campylobacter species. Based on MLST, 70.9% of the human cases were attributed to chickens, 19.3% to cattle, 8.6% to dogs and 1.2% to pigs. Furthermore we found a host independent association between sequence type (ST) and quinolone resistance. The most notable were ST-45, all isolates of which were susceptible, while for ST-464 all were resistant. PMID:24244747

  3. Source attribution of human Campylobacter isolates by MLST and fla-typing and association of genotypes with quinolone resistance.

    Directory of Open Access Journals (Sweden)

    Sonja Kittl

    Full Text Available Campylobacteriosis is the most frequent zoonosis in developed countries and various domestic animals can function as reservoir for the main pathogens Campylobacter jejuni and Campylobacter coli. In the present study we compared population structures of 730 C. jejuni and C. coli from human cases, 610 chicken, 159 dog, 360 pig and 23 cattle isolates collected between 2001 and 2012 in Switzerland. All isolates had been typed with multi locus sequence typing (MLST and flaB-typing and their genotypic resistance to quinolones was determined. We used complementary approaches by testing for differences between isolates from different hosts with the proportion similarity as well as the fixation index and by attributing the source of the human isolates with Bayesian assignment using the software STRUCTURE. Analyses were done with MLST and flaB data in parallel and both typing methods were tested for associations of genotypes with quinolone resistance. Results obtained with MLST and flaB data corresponded remarkably well, both indicating chickens as the main source for human infection for both Campylobacter species. Based on MLST, 70.9% of the human cases were attributed to chickens, 19.3% to cattle, 8.6% to dogs and 1.2% to pigs. Furthermore we found a host independent association between sequence type (ST and quinolone resistance. The most notable were ST-45, all isolates of which were susceptible, while for ST-464 all were resistant.

  4. Real-Time PCR using a PCR Microchip with Integrated Thermal System and Polymer Waveguides for the Detection of Campylobacter jejuni

    DEFF Research Database (Denmark)

    Wang, Zhenyu; Sekulovic, Andrea; Kutter, Jörg Peter;

    2006-01-01

    A novel real-time PCR microchip platform with integrated thermal system and polymer waveguides has been developed. By using the integrated optical system of the real-time PCR chip, cadF – a virulence gene of Campylobacter jejuni, could specifically be detected. Two different DNA binding dyes, SYTOX...

  5. 空肠弯曲菌毒力因子研究进展%Overview on Virulence Factors of Campylobacter jejuni

    Institute of Scientific and Technical Information of China (English)

    钱卫东; 王婷; 施春阳

    2011-01-01

    空肠弯曲菌(Campylobacter jejuni)是世界范围内广泛流行的人畜共患病,在公共卫生学上具有重要意义.空肠弯曲菌可引发人和多种动物疾病,其抗原结构复杂,致病机理尚不清楚.作者通过对空肠弯曲菌可能的毒力因子及其在致病中的作用作一简要概述,为空肠弯曲菌的诊断防治提供依据.%Campylobacter jejuni is the principal zoonotic pathogen of global concern and widely perceived as an increasing serious public health problem. Campylobacter jejuni may cause many diseases in animal and human,but the structure of antigen and the exact pathogenesis are still not clear. This paper reviewes the possible pathogenesis-associated factors and disease-mediating mechanisms,which provides a basis for epidemiologic investigation and prevention of Campylobacter jejuni.

  6. The Campylobacter jejuni RacRS two-component system activates the glutamate synthesis by directly upregulating γ-glutamyltranspeptidase (GGT)

    NARCIS (Netherlands)

    van der Stel, Anne-Xander; van Mourik, Andries; Łaniewski, Paweł; van Putten, Jos P M; Jagusztyn-Krynicka, Elżbieta K; Wosten, Marc

    2015-01-01

    The highly conserved enzyme γ-glutamyltranspeptidase (GGT) plays an important role in metabolism of glutathione and glutamine. Yet, the regulation of ggt transcription in prokaryotes is poorly understood. In the human pathogen Campylobacter jejuni, GGT is important as it contributes to persistent co

  7. Inter-laboratory evaluation of three flagellin PCR/RFLP methods for typing Campylobacter jejuni and C-coli: the CAMPYNET experience

    DEFF Research Database (Denmark)

    Harrington, Clare S.; Moran, L.; Ridley, A.M.;

    2003-01-01

    Aims: To compare typeability, discriminatory ability, and inter-laboratory reproducibility of three flagellin PCR/RFLP(fla typing) methods previously described for Campylobacter. Methods and Results: The sample set(n = 100) was diverse, including both C. jejuni (n = 85) and C. coli (n = 15). Two...

  8. The influence of feeding crimped kernel maize silage on growth performance and intestinal colonization with Campylobacter jejuni in broilers

    DEFF Research Database (Denmark)

    Ranjitkar, Samir; Engberg, Ricarda Greuel

    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% an...

  9. Crystal Structure of the Transcriptional Regulator CmeR From Campylobacter Jejuni

    Energy Technology Data Exchange (ETDEWEB)

    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.

  10. Cepas de Campylobacter jejuni resistentes a quinolonas aisladas de humanos, gallinas y pollos

    Directory of Open Access Journals (Sweden)

    Rodolfo Notario

    2011-08-01

    Full Text Available Se compararon 8 aislamientos de Campylobacter jejuni provenientes de humanos con enfermedad diarreica aguda, con 23 aislamientos de cloaca de gallinas y pollos obtenidos de zonas próximas a la ciudad de Rosario, todos resistentes a la ciprofloxacina. Las muestras se sembraron en agar selectivo y se incubaron en microaerofilia a 42 °C. Las colonias se identificaron con el método tradicional. Los aislamientos se conservaron a -70 °C en caldo cerebro corazón con 17% v/v de glicerina. La clonalidad se determinó por RAPD-PCR, utilizando el primer 1254 (Stern NJ. Se interpretaron los aislamientos como clones distintos cuando diferían en una banda de amplificación. Se obtuvieron 5 clones diferentes. Los patrones I, II y V fueron aislados en criaderos industriales de pollos y en humanos (el II también en un establecimiento de gallinas ponedoras de huevos. En un gallinero familiar se obtuvo el patrón I. El patrón III sólo se obtuvo de humanos. El patrón IV se halló en uno de los criaderos pero no en humanos. Se pudo determinar que 93.5% de las cepas se aislaron tanto de animales como de humanos, por lo que se considera posible que la colonización de criaderos con cepas resistentes a los antimicrobianos pudiera ser el origen de la infección de humanos.

  11. Antimicrobial Resistance Profiles of Campylobacter spp. Isolated from Broiler Chicken Meat of Estonian, Latvian and Lithuanian Origin at Estonian Retail Level and from Patients with Severe Enteric Infections in Estonia.

    Science.gov (United States)

    Mäesaar, M; Kramarenko, T; Meremäe, K; Sõgel, J; Lillenberg, M; Häkkinen, L; Ivanova, M; Kovalenko, K; Hörman, A; Hänninen, M-L; Roasto, M

    2016-03-01

    The resistance patterns of Campylobacter spp. isolated from retail broiler chicken meat originating either from Estonia, Lithuania or Latvia collected in Estonia were determined. Additionally, in collaboration with the laboratories of several Estonian hospitals, antimicrobial susceptibility patterns were determined for Campylobacter isolates from patients with severe Campylobacter enteric infections. The isolates were identified at the species level by the PCR method. Respectively, 88.8% of the isolates were C. jejuni, and 11.2% were C. coli. In total, 126 Campylobacter isolates of broiler chicken meat and human origin were tested for minimal inhibitory concentrations (MICs) with the broth microdilution VetMIC(TH) method (National Veterinary Institute; Uppsala, Sweden) for a total of six antimicrobials. Resistance to one or more antimicrobials was detected in 62 (63.3%) of Campylobacter broiler chicken meat isolates and in 20 (71.4%) of human-origin isolates. Large proportions of the broiler chicken meat isolates were resistant to ciprofloxacin (60.2%). Multidrug resistance (i.e. to three or more unrelated antimicrobials) was detected in five (5.1%) C. jejuni isolates. Among the human isolates, 20 (71.4%) were resistant to fluoroquinolones, and two (7.1%) C. jejuni isolates exhibited multidrug resistance. The chicken meat isolates of Estonian origin were the most susceptible. However, a high proportion of fluoroquinolone-resistant C. jejuni isolates were found in Latvian and Lithuanian products. The results of this study indicate that the problems caused by the inappropriate use of antimicrobials extend beyond the country in which a food originates; therefore, both domestic and international interventions and agreements are required to implement common policies on antimicrobial usage and to minimize the emergence of Campylobacter drug resistance.

  12. Prevalence and characterization of Campylobacter spp. isolated from domestic and imported poultry meat in Korea, 2004–2008.

    Science.gov (United States)

    Kim, Hae Ji; Kim, Jong Hyun; Kim, Young Ihl; Choi, Jung Su; Park, Mi Young; Nam, Hyang-Mi; Jung, Suk-Chan; Kwon, Jin Wook; Lee, Chul Hyun; Kim, Yong Hwan; Ku, Bok Kyung; Lee, Young Ju

    2010-10-01

    Campylobacteriosis in humans is primarily caused by handling or consuming contaminated poultry or their products. The aims of this study were to estimate the prevalence of Campylobacter spp. in domestic and imported poultry meat in Korea and to further characterize the obtained isolates. From 2004 to 2008, a total of 475 domestic and 867 imported raw poultry meat samples were examined for the presence of Campylobacter spp. Among 475 domestic poultry meat samples, Campylobacter jejuni and Campylobacter coli were isolated from 219 (46.1%) and 156 (32.8%), respectively. Relative prevalence of C. jejuni and C. coli was higher in meat from Brazil (39/78, 50.0% and 7/78, 8.9%) and France (32/96, 33.3% and 8/96, 8.3%), whereas lower in meat from Denmark (72/516, 14.0% and 12/516, 2.3%) and Thailand (5/39, 12.8% and 3/39, 7.6%). The resistance to ampicillin and tetracycline was highly prevalent in Campylobacter spp. from most countries investigated, whereas lower in meat from Denmark. On the other hand, the prevalence of erythromycin and gentamicin resistance was less than 10% in most countries. The resistance rate to nalidixic acid, ciprofloxacin, and enrofloxacin ranged from 11.9% to 87.5%. The use of fla-polymerase chain reaction–restriction fragment length polymorphism for epidemiological analysis found that some pattern types were considerably more frequent and distinct in meat from each country. In conclusion, we report the presence of high contamination in domestic and imported poultry meat in Korea and the antimicrobial and genetic diversity of Campylobacter spp. between each country.

  13. A comparative study of two food model systems to test the survival of Campylobacter jejuni at -18 degrees C

    DEFF Research Database (Denmark)

    Birk, Tina; Rosenquist, Hanne; Brondsted, L.;

    2006-01-01

    The survival of Campylobacter jejuni NCTC 11168 was tested at freezing conditions (-18 degrees C) over a period of 32 days in two food models that simulated either (i) the chicken skin surface (skin model) or (ii) the chicken juice in and around a broiler carcass (liquid model). In the skin model...... NCTC 11168 cells was slower when suspended in chicken juice than in BHIB. After freezing for 32 days, the reductions in the cell counts were 1.5 log CFU/ml in chicken juice and 3.5 log CFU/ml in BHIB. After the same time of freezing but when inoculated onto chicken skin, C. jejuni NCTC 11168...... was reduced by 2.2 log units when inoculated in chicken juice and 3.2 log units when inoculated into BHIB. For both models, the major decrease occurred within the first 24 h of freezing. The results obtained in the liquid model with chicken juice were comparable to the reductions of Campylobacter observed...

  14. The Campylobacter jejuni CiaC virulence protein is secreted from the flagellum and delivered to the cytosol of host cells

    Directory of Open Access Journals (Sweden)

    Jason eNeal-McKinney

    2012-03-01

    Full Text Available Campylobacter jejuni is a leading cause of bacterial gastroenteritis worldwide. Acute C. jejuni-mediated disease (campylobacteriosis involves C. jejuni invasion of host epithelial cells using a set of virulence proteins known as the Campylobacter invasion antigens (Cia. The genes encoding the Cia proteins are up-regulated upon co-culture of C. jejuni with epithelial cells. One of the Cia proteins, CiaC, is required for maximal invasion of host cells by C. jejuni. Previous work has also revealed that CiaC is, in part, responsible for host cell cytoskeletal rearrangements that result in membrane ruffling. This study was performed to test the hypothesis that CiaC is delivered to the cytosol of host cells. To detect the delivery of CiaC into cultured epithelial cells, we used the adenylate cyclase domain (ACD of Bordetella pertussis CyaA as a reporter. In this study, we found that export and delivery of the C. jejuni Cia proteins into human INT 407 epithelial cells required a functional flagellar hook complex composed of FlgE, FlgK, and FlgL. Assays performed with bacterial culture supernatants supported the hypothesis that CiaC delivery requires bacteria-host cell contact. We also found that that CiaC was delivered to host cells by cell-associated (bound bacteria, as judged by experiments performed with inhibitors that specifically target the cell signaling pathways utilized by C. jejuni for cell invasion. Interestingly, the C. jejuni flgL mutant, which is incapable of exporting and delivering the Cia proteins, did not induce INT 407 cell membrane ruffles. Complementation of the flgL mutant with plasmid-encoded flgL restored the motility and membrane ruffling. These data support the hypothesis that the C. jejuni Cia proteins, which are exported from the flagellum, are delivered to the cytosol of host cells.

  15. Eugenol wash and chitosan based coating reduces Campylobacter jejuni counts on poultry products

    Science.gov (United States)

    Campylobacter, a leading cause of foodborne illness globally in humans, is strongly associated with the consumption of contaminated poultry products. Unfortunately, current strategies to reduce Campylobacter counts in poultry have had limited success. Our study investigated the efficacy of eugenol ...

  16. Effect of steam and lactic acid treatments on the survival of Salmonella Enteritidis and Campylobacter jejuni inoculated on chicken skin.

    Science.gov (United States)

    Chaine, Aline; Arnaud, Elodie; Kondjoyan, Alain; Collignan, Antoine; Sarter, Samira

    2013-04-01

    Campylobacteriosis and salmonellosis are the most frequently reported zoonotic infectious diseases. The present work evaluated the effectiveness of steam treatment at 100 °C for 8s, a 5% lactic acid treatment for 1 min and their combination for inactivating Salmonella Enteritidis and Campylobacter jejuni inoculated on chicken skin. The impact of each treatment on the total aerobic mesophilic bacteria and the effect of rinsing after contact with lactic acid were also evaluated. Residual bacteria were counted immediately after treatment or after seven days of storage at 4 °C. Results demonstrated the immediate efficiency of the steam and the combined treatments with reductions of approximately 6 and 5 log cfu/cm2 respectively for S. Enteritidis and C. jejuni. They also showed significant reductions (equal to or >3.2 log cfu/cm2) in the total aerobic mesophilic plate count. Lactic acid had a persistent effect on pathogen growth during storage which was significantly higher when the skin was not rinsed, reaching reductions of 3.8 log cfu/cm2 for both S. Enteritidis and C. jejuni. Only the combined treatments significantly reduced the recovery of the total aerobic mesophilic bacteria during storage. The significant reductions in both pathogens and total aerobic mesophilic bacteria on treated chicken skins are possible ways to improve the safety and shelf life of the product although high levels of indigenous non-pathogenic bacteria may be beneficial due to their protective effect against potential re-contamination of chicken skin.

  17. Polar flagellar biosynthesis and a regulator of flagellar number influence spatial parameters of cell division in Campylobacter jejuni.

    Directory of Open Access Journals (Sweden)

    Murat Balaban

    2011-12-01

    Full Text Available Spatial and numerical regulation of flagellar biosynthesis results in different flagellation patterns specific for each bacterial species. Campylobacter jejuni produces amphitrichous (bipolar flagella to result in a single flagellum at both poles. These flagella confer swimming motility and a distinctive darting motility necessary for infection of humans to cause diarrheal disease and animals to promote commensalism. In addition to flagellation, symmetrical cell division is spatially regulated so that the divisome forms near the cellular midpoint. We have identified an unprecedented system for spatially regulating cell division in C. jejuni composed by FlhG, a regulator of flagellar number in polar flagellates, and components of amphitrichous flagella. Similar to its role in other polarly-flagellated bacteria, we found that FlhG regulates flagellar biosynthesis to limit poles of C. jejuni to one flagellum. Furthermore, we discovered that FlhG negatively influences the ability of FtsZ to initiate cell division. Through analysis of specific flagellar mutants, we discovered that components of the motor and switch complex of amphitrichous flagella are required with FlhG to specifically inhibit division at poles. Without FlhG or specific motor and switch complex proteins, cell division occurs more often at polar regions to form minicells. Our findings suggest a new understanding for the biological requirement of the amphitrichous flagellation pattern in bacteria that extend beyond motility, virulence, and colonization. We propose that amphitrichous bacteria such as Campylobacter species advantageously exploit placement of flagella at both poles to spatially regulate an FlhG-dependent mechanism to inhibit polar cell division, thereby encouraging symmetrical cell division to generate the greatest number of viable offspring. Furthermore, we found that other polarly-flagellated bacteria produce FlhG proteins that influence cell division, suggesting that

  18. Rapid PCR using nested primers of the 16S rRNA and the hippuricase (hipO) genes to detect Campylobacter jejuni and Campylobacter coli in environmental samples

    DEFF Research Database (Denmark)

    Bang, Dang Duong; Wedderkopp, A.; Pedersen, Karl;

    2002-01-01

    Identification of sources Campylobacter infection in the poultry houses is in general problematic due to the lack of reliable methods to detect campylobacteria in environmental samples. Detection of campylobacteria in environmental samples by conventional culture methods is difficult and of limited...... sensitivity due to the use of selective media, the low number of bacteria in the samples and possibly also due to the presence of non-culturable or sub-lethally injured stages of the bacteria. The present paper describes a rapid PCR assay using nested primers of the 16S rRNA or the hippuricase (hipO) genes...... to detect Campylobacter jejuni and Campylobacter coli in environmental samples. The sensitivity of the nested PCR was determined to be 0.01 pg/PCR, corresponding to 2-3 colony forming units (cfu) per ml. The nested PCR assays were applied to detect C. jejuni and C. coli in 269 environmental samples...

  19. Identificación mediante biotipos y perfiles proteicos de Campylobacter aislados de perros Identification of Campylobacters isolated from dogs by using biotypes and protein electrophoretic patterns

    Directory of Open Access Journals (Sweden)

    G. GIACOBONI

    1999-01-01

    Full Text Available Se estudiaron 260 muestras fecales obtenidas por hisopado rectal atendidos en la Facultad de Ciencias Veterinarias de la Universidad Nacional de La Plata. Para el aislamiento de Campylobacter se utilizó caldo Preston de enriquecimiento, incubado a 43°C durante 12 h en microaerofilia y siembra en agar Skirrow incubado en iguales condiciones por 48 h. La identificación de especie se realizó por los métodos convencionales, la biotipificación por el método de Lior y los perfiles proteicos se determinaron en geles de poliacrilamida bajo condiciones de reducción. La prevalencia de Campylobacter termotolerantes fue de 7.3%, resultando 19 cepas identificadas como C. jejuni biotipo II. De éstas, 15 fueron sometidas a electroforesis, utilizándose la cepa humana 1522 como referencia. Los perfiles proteicos obtenidos de las cepas fueron similares y presentaron un número de 30 bandas aproximadamente. Todas las cepas compartieron las bandas del polipéptido mayor de 40-46 kD. Las diferencias se observaron en la carencia de algunas bandas o en su movilidad entre las de 40 y 50 kD. El hecho de que la electroforesis de proteína haya demostrado heterogeneidad entre cepas de una misma especie y biotipo, reafirma su importancia como método de discriminación epidemiológicaTwo hundred and sixty faecal samples obtained with rectal swabs from dogs were studied for Campylobacter isolation. The samples were placed on Preston broth and incubated at 43° C for 12 h under a microaerophilic atmosphere. Then, they were seeded on Skirrow agar and incubated under the same conditions for 48 h. Campylobacter species were identified by conventional methods (morphological, biochemical and cultural test, Lior's biotypes and protein profiles were examined by polyacrylamide gel electrophoresis, under reducing conditions. The prevalence of thermo-tolerant Campylobacter species was 7.3%. Fifteen out of 19 Campylobacter jejuni biotype II isolates were used in

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

    Science.gov (United States)

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

    2016-01-01

    Campylobacter jejuni is a leading cause of bacterial gastroenteritis in the developed world. Despite its prevalence, its mechanisms of pathogenesis are poorly understood. Peptidoglycan (PG) is important for helical shape, colonization, and host-pathogen interactions in C. jejuni. Therefore, changes in PG greatly impact the physiology of this organism. O-acetylation of peptidoglycan (OAP) is a bacterial phenomenon proposed to be important for proper cell growth, characterized by acetylation of the C6 hydroxyl group of N-acetylmuramic acid in the PG glycan backbone. The OAP gene cluster consists of a PG O-acetyltransferase A (patA) for translocation of acetate into the periplasm, a PG O-acetyltransferase B (patB) for O-acetylation, and an O-acetylpeptidoglycan esterase (ape1) for de-O-acetylation. In this study, reduced OAP in ΔpatA and ΔpatB had minimal impact on C. jejuni growth and fitness under the conditions tested. However, accumulation of OAP in Δape1 resulted in marked differences in PG biochemistry, including O-acetylation, anhydromuropeptide levels, and changes not expected to result directly from Ape1 activity. This suggests that OAP may be a form of substrate level regulation in PG biosynthesis. Ape1 acetylesterase activity was confirmed in vitro using p-nitrophenyl acetate and O-acetylated PG as substrates. In addition, Δape1 exhibited defects in pathogenesis-associated phenotypes, including cell shape, motility, biofilm formation, cell surface hydrophobicity, and sodium deoxycholate sensitivity. Δape1 was also impaired for chick colonization and adhesion, invasion, intracellular survival, and induction of IL-8 production in INT407 cells in vitro. The importance of Ape1 in C. jejuni biology makes it a good candidate as an antimicrobial target. PMID:27474744

  1. A Designed Experiments Approach to Optimizing MALDI-TOF MS Spectrum Processing Parameters Enhances Detection of Antibiotic Resistance in Campylobacter jejuni.

    Science.gov (United States)

    Penny, Christian; Grothendick, Beau; Zhang, Lin; Borror, Connie M; Barbano, Duane; Cornelius, Angela J; Gilpin, Brent J; Fagerquist, Clifton K; Zaragoza, William J; Jay-Russell, Michele T; Lastovica, Albert J; Ragimbeau, Catherine; Cauchie, Henry-Michel; Sandrin, Todd R

    2016-01-01

    MALDI-TOF MS has been utilized as a reliable and rapid tool for microbial fingerprinting at the genus and species levels. Recently, there has been keen interest in using MALDI-TOF MS beyond the genus and species levels to rapidly identify antibiotic resistant strains of bacteria. The purpose of this study was to enhance strain level resolution for Campylobacter jejuni through the optimization of spectrum processing parameters using a series of designed experiments. A collection of 172 strains of C. jejuni were collected from Luxembourg, New Zealand, North America, and South Africa, consisting of four groups of antibiotic resistant isolates. The groups included: (1) 65 strains resistant to cefoperazone (2) 26 resistant to cefoperazone and beta-lactams (3) 5 strains resistant to cefoperazone, beta-lactams, and tetracycline, and (4) 76 strains resistant to cefoperazone, teicoplanin, amphotericin, B and cephalothin. Initially, a model set of 16 strains (three biological replicates and three technical replicates per isolate, yielding a total of 144 spectra) of C. jejuni was subjected to each designed experiment to enhance detection of antibiotic resistance. The most optimal parameters were applied to the larger collection of 172 isolates (two biological replicates and three technical replicates per isolate, yielding a total of 1,031 spectra). We observed an increase in antibiotic resistance detection whenever either a curve based similarity coefficient (Pearson or ranked Pearson) was applied rather than a peak based (Dice) and/or the optimized preprocessing parameters were applied. Increases in antimicrobial resistance detection were scored using the jackknife maximum similarity technique following cluster analysis. From the first four groups of antibiotic resistant isolates, the optimized preprocessing parameters increased detection respective to the aforementioned groups by: (1) 5% (2) 9% (3) 10%, and (4) 2%. An additional second categorization was created from the

  2. A Designed Experiments Approach to Optimizing MALDI-TOF MS Spectrum Processing Parameters Enhances Detection of Antibiotic Resistance in Campylobacter jejuni

    Directory of Open Access Journals (Sweden)

    Christian ePenny

    2016-05-01

    Full Text Available MALDI-TOF MS has been utilized as a reliable and rapid tool for microbial fingerprinting at the genus and species levels. Recently, there has been keen interest in using MALDI-TOF MS beyond the genus and species levels to rapidly identify antibiotic resistant strains of bacteria. The purpose of this study was to enhance strain level resolution for Campylobacter jejuni through the optimization of spectrum processing parameters using a series of designed experiments. A collection of 172 strains of C. jejuni were collected from Luxembourg, New Zealand, North America, and South Africa, consisting of four groups of antibiotic resistant isolates. The groups included: 1 65 strains resistant to cefoperazone 2 26 resistant to cefoperazone and beta-lactams 3 5 strains resistant to cefoperazone, beta-lactams, and tetracycline, and 4 76 strains resistant to cefoperazone, teicoplanin, amphotericin B and cephalothin. Initially, a model set of 16 strains (three biological replicates and three technical replicates per isolate, yielding a total of 144 spectra of C. jejuni was subjected to each designed experiment to enhance detection of antibiotic resistance. The most optimal parameters were applied to the larger collection of 172 isolates (two biological replicates and three technical replicates per isolate, yielding a total of 1,031 spectra. We observed an increase in antibiotic resistance detection whenever either a curve based similarity coefficient (Pearson or ranked Pearson was applied rather than a peak based (Dice and/or the optimized preprocessing parameters were applied. Increases in antimicrobial resistance detection were scored using the jackknife maximum similarity technique following cluster analysis. From the first four groups of antibiotic resistant isolates, the optimized preprocessing parameters increased detection respective to the aforementioned groups by: 1 five percent 2 nine percent 3 ten percent, and 4 two percent. An additional second

  3. Nitrosylation mechanisms of Mycobacterium tuberculosis and Campylobacter jejuni truncated hemoglobins N, O, and P.

    Directory of Open Access Journals (Sweden)

    Paolo Ascenzi

    Full Text Available Truncated hemoglobins (trHbs are widely distributed in bacteria and plants and have been found in some unicellular eukaryotes. Phylogenetic analysis based on protein sequences shows that trHbs branch into three groups, designated N (or I, O (or II, and P (or III. Most trHbs are involved in the O2/NO chemistry and/or oxidation/reduction function, permitting the survival of the microorganism in the host. Here, a detailed comparative analysis of kinetics and/or thermodynamics of (i ferrous Mycobacterium tuberculosis trHbs N and O (Mt-trHbN and Mt-trHbO, respectively, and Campylobacter jejuni trHb (Cj-trHbP nitrosylation, (ii nitrite-mediated nitrosylation of ferrous Mt-trHbN, Mt-trHbO, and Cj-trHbP, and (iii NO-based reductive nitrosylation of ferric Mt-trHbN, Mt-trHbO, and Cj-trHbP is reported. Ferrous and ferric Mt-trHbN and Cj-trHbP display a very high reactivity towards NO; however, the conversion of nitrite to NO is facilitated primarily by ferrous Mt-trHbN. Values of kinetic and/or thermodynamic parameters reflect specific trHb structural features, such as the ligand diffusion pathways to/from the heme, the heme distal pocket structure and polarity, and the ligand stabilization mechanisms. In particular, the high reactivity of Mt-trHbN and Cj-trHbP reflects the great ligand accessibility to the heme center by two protein matrix tunnels and the E7-path, respectively, and the penta-coordination of the heme-Fe atom. In contrast, the heme-Fe atom of Mt-trHbO the ligand accessibility to the heme center of Mt-trHbO needs large conformational readjustments, thus limiting the heme-based reactivity. These results agree with different roles of Mt-trHbN, Mt-trHbO, and Cj-trHbP in vivo.

  4. Campylobacter jejuni acquire new host-derived CRISPR spacers when in association with bacteriophages harboring a CRISPR-like Cas4 protein

    OpenAIRE

    Hooton, Steven P. T.; Connerton, I.F.

    2015-01-01

    Campylobacter jejuni is a worldwide cause of human diarrhoeal disease. Clustered Repetitively Interspaced Palindromic Repeats (CRISPRs) and associated proteins allow Bacteria and Archaea to evade bacteriophage and plasmid infection. Type II CRISPR systems are found in association with combinations of genes encoding the CRISPR-associated Cas1, Cas2, Cas4 or Csn2, and Cas9 proteins. C. jejuni possesses a minimal subtype II-C CRISPR system containing cas1, cas2, and cas9 genes whilst cas4 is not...

  5. 空肠弯曲菌和沙门菌双重实时荧光定量 PCR 检测方法的建立%Duplex Real-time PCR for Simultaneous Detection of Campylobacter jejuni and Salmonella

    Institute of Scientific and Technical Information of China (English)

    高瑞娟; 张凯; 吕嘉敏; 黄永兴; 罗开健

    2014-01-01

    This study aimed to establish a rapid detection of Campylobacter jejuni and Salmonella by duplex real-time PCR.The primers and Taq man probes were designed based on the invA gene of Salmonella and hipO gene of Campylobacter jejuni ,using FAM,JOE and TAMRA fluorescently labeled specific probes aimed at the conserved genes of target pathogens,respectively.We compared Tm values to fix these primers and probes,and optimize the reaction conditions to establish the duplex real-time PCR approach for the detection of Campylobacter jejuni and Salmonella.The sensitivity of the duplex real-time PCR was 10 CFU/mL,230 CFU/mL for Campylobacterjejuni and Salmonella.The duplex real-time PCR showed good specificity which was validated by testing various bacteria isolates.In this study,a duplex PCR with high specificity and sensitivity was developed and it would provide useful information for the simultaneous detection of Campylobacter jejuni and Salmonella in food.%为建立一种空肠弯曲菌和沙门菌的双重实时荧光定量 PCR 检测方法,根据空肠弯曲菌的保守基因 hipO 和沙门菌的保守基因 invA 分别设计合成引物和 TaqMan 探针,分别使用 FAM、JOE 作为探针报告基团,TAMRA 作为探针淬灭基团。优化反应体系及条件,建立适用于检测食品中空肠弯曲菌和沙门菌的双重实时荧光定量 PCR 方法。结果显示,该检测方法特异性强,灵敏度高,空肠弯曲菌检测限可达10 CFU/mL,沙门菌检测限达230 CFU/mL。表明建立的双重实时荧光定量 PCR 可为实现食品中空肠弯曲菌和沙门菌的同时检测提供新方法。

  6. Increased risk for Campylobacter jejuni and C. coli infection of pet origin in dog owners and evidence for genetic association between strains causing infection in humans and their pets

    NARCIS (Netherlands)

    Gras, L.M.; Smid, J.H.; Wagenaar, J.A.; Koene, M.G.J.; Havelaar, A.H.; Friesema, I.H.M.; French, N.P.; Flemming, C.; Galson, J.D.; Graziani, C.; Busani, L.; Pelt, van W.

    2013-01-01

    We compared Campylobacter jejuni/coli multilocus sequence types (STs) from pets (dogs/cats) and their owners and investigated risk factors for pet-associated human campylobacteriosis using a combined source-attribution and case-control analysis. In total, 132/687 pet stools were Campylobacter-positi

  7. Research progress on flagella of Campylobacter jejuni%空肠弯曲菌鞭毛研究进展

    Institute of Scientific and Technical Information of China (English)

    任方哲; 王楠; 商宇伟; 焦新安; 黄金林

    2013-01-01

    Campylobacter jejuni is a food-borne bacterial pathogen popular worldwide.Flagella is the main virulence factor for this bacteria and plays an important role in its pathogenesis.In this paper,the flagellar structure,biogenesis process and function is reviewed to provide a reference for further understanding the flagellar basic information and pathogenesis mechanisms of Campylobacterjejuni.%空肠弯曲菌是世界范围内广泛流行的一种食源性病原菌,鞭毛是其主要的毒力因子,在其致病过程中发挥重要作用.本文对空肠弯曲菌鞭毛结构,合成机制及其作用进行综述,以期为了解空肠弯曲菌鞭毛的基本信息及其致病机理提供参考.

  8. Structural Context for Protein N-glycosylation in Bacteria: The Structure of PEB3, an Adhesin from Campylobacter Jejuni

    Energy Technology Data Exchange (ETDEWEB)

    Rangarajan,E.; Bhatia, S.; Watson, D.; Munger, C.; Cygler, M.; Matte, A.; Young, N.

    2007-01-01

    Campylobacter jejuni is unusual among bacteria in possessing a eukaryotic-like system for N-linked protein glycosylation at Asn residues in sequons of the type Asp/Glu-Xaa-Asn-Xaa-Ser/Thr. However, little is known about the structural context of the glycosylated sequons, limiting the design of novel recombinant glycoproteins. To obtain more information on sequon structure, we have determined the crystal structure of the PEB3 (Cj0289c) dimer. PEB3 has the class II periplasmic-binding protein fold, with each monomer having two domains with a ligand-binding site containing citrate located between them, and overall resembles molybdate- and sulfate-binding proteins. The sequon around Asn90 is located within a surface-exposed loop joining two structural elements. The three key residues are well exposed on the surface; hence, they may be accessible to the PglB oligosaccharyltransferase in the folded state.

  9. Longitudinal study of the excretion patterns of thermophilic Campylobacter spp. in young pet dogs in Denmark

    DEFF Research Database (Denmark)

    Hald, Birthe; Pedersen, Karl; Wainø, Michael;

    2004-01-01

    The Campylobacter excretion patterns of 26 domestic pet dogs were described in a longitudinal study. The dogs entered the study between 3 and 8 months of age and were monitored until 2 years of age. They were tested monthly for Campylobacter carriage in stool samples that were cultured...... on the Campylobacter-selective media CAT and modified CCDA agar at 37 and 42 C. This study comprised 366 fecal swab samples, of which 278 (76.2%) were found to be Campylobacter positive, with the following distribution of species: 75.0% Campylobacter upsaliensis, 19.4% Campylobacter jejuni, 2.1% Campylobacter lari, 0.......7% Campylobacter coli, and 2.8% Campylobacter spp. Isolates were typed by pulsed-field gel electrophoresis (PFGE) to elucidate the strain excretion pattern. All study dogs excreted Campylobacter spp. during the study period. At 3 months of age, 60% of the dogs carried Campylobacter, increasing to nearly 100...

  10. Serotipos de Campylobacter jejuni ssp. jejuni aislado en carne de ave para consumo humano y en muestras de heces de niños con diarrea Campylobacter jejuni ssp. jejuni serotypes in avian meat for human consumption and in faecal samples of children with diarrhoea

    Directory of Open Access Journals (Sweden)

    H. Fernández

    2005-01-01

    Full Text Available Considerando a las aves de consumo como un importante reservorio de C. jejuni ssp jejuni y sus subproductos alimenticios como principal vehículo, fueron serotipificadas 50 cepas aisladas de carne de ave para consumo humano, correlacionándolas epidemiológicamente con los serotipos encontrados en 50 cepas aisladas de niños con diarrea. Las cepas de C.jejuni ssp jejuni fueron serotipificadas mediante la técnica de hemaglutinación pasiva descrita por Penner, utilizando un kit comercial (DENKA SEIKEN, conteniendo los 25 sueros tipificadores para los serotipos descritos como los más frecuentes. Los serotipos A, B, F, L, N e Y fueron aislados concomitantemente en ambos tipos de muestras. Los serotipos de mayor aislamiento en las muestras de origen humano fueron Z2 (16% y Z5 (12%, mientras que en las de origen aviar fueron los serotipos A (28%, C (10%, L (10% e Y (10%. Al no coexistir los serotipos Z y C, en ambos grupos muestrales, es posible inferir la existencia de otros reservorios y vehículos que estén actuando como agentes transmisores de esta bacteria al ser humano.Having in mind that fowl and avian byproducts for human consumption are important reservoirs and vehicles for C. jejuni ssp. jejuni, a serotyping study was conducted in 50 strains isolated from avian meat and 50 isolated from human diarrhoeic stools. C. jejuni ssp. jejuni serotyping was carried out using a commercial kit with 25 antisera (DENKA SEIKEN, based on the Penner’s passive haemagglutination technique. Serotypes A, B, F, L, N e Y were concomitantly found in both kinds of samples. Serotypes Z2 (16% and Z5 (12% were the most frequently found in human strains, whereas serotypes A (28%, C (10%, L (10% and Y (10% were the most frequently isolated from avian samples. However, serotypes Z and C did not coexist in both types of samples. The latter results suggest that additionally to fowl and avian meat, there might be other animal reservoirs and vehicles that could act as

  11. Phenotypic and Genotypic Evidence for L-fucose Utilization by Campylobacter jejuni

    Science.gov (United States)

    Campylobacter 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....

  12. Prevalence of Campylobacter jejuni and Campylobacter coli in Market-Weight Turkeys On-Farm and at Slaughter

    Science.gov (United States)

    To monitor the effects of feed withdrawal on the prevalence of Campylobacter, market weight turkeys from six farms were examined before and after perimarketing events (feed withdrawal, transport, and holding at the slaughterhouse). Prior to transport, birds (n = 30/farm) were slaughtered on-farm an...

  13. Antimicrobial, Rheological, and Thermal Properties of Plasticized Polylactide Films Incorporated with Essential Oils to Inhibit Staphylococcus aureus and Campylobacter jejuni.

    Science.gov (United States)

    Ahmed, Jasim; Hiremath, Nikhil; Jacob, Harsha

    2016-02-01

    Polylactide (PLA) is the most mature biobased and biodegradable polymer. Due to its inherent brittleness, the polymer cannot be used as a packaging material without plasticizer. An attempt was made to develop antimicrobial plasticized PLA film by incorporating polyethylene glycol (PEG) and 3 essential oils (EO), namely cinnamon, garlic, and clove by solvent casting method. Physical, thermal, and rheological properties of those films were evaluated for practical applications whereas the antimicrobial properties were tested against Staphylococcus aureus and Campylobacter jejuni-pathogens related to poultry industry. Both PEG and EOs led to the formation of flexible PLA/PEG/EO films with significant drop in the glass transition temperature (Tg ), and mechanical property. Time-temperature superposition (TTS) principle was employed to melt rheology of EO-based films at selected temperature, and rheological moduli superimposed well in an extended frequency range. Among EOs, cinnamon and clove oil-based films (PLA/PEG/CIN and PLA/PEG/CLO) exhibited a complete zone of inhibition against C. jejuni at the maximum concentration (1.6 mL per 2 g PLA/PEG blend) whereas the garlic oil-based film (PLA/PEG/GAR) had the lowest activity. PMID:26749466

  14. Antimicrobial, Rheological, and Thermal Properties of Plasticized Polylactide Films Incorporated with Essential Oils to Inhibit Staphylococcus aureus and Campylobacter jejuni.

    Science.gov (United States)

    Ahmed, Jasim; Hiremath, Nikhil; Jacob, Harsha

    2016-02-01

    Polylactide (PLA) is the most mature biobased and biodegradable polymer. Due to its inherent brittleness, the polymer cannot be used as a packaging material without plasticizer. An attempt was made to develop antimicrobial plasticized PLA film by incorporating polyethylene glycol (PEG) and 3 essential oils (EO), namely cinnamon, garlic, and clove by solvent casting method. Physical, thermal, and rheological properties of those films were evaluated for practical applications whereas the antimicrobial properties were tested against Staphylococcus aureus and Campylobacter jejuni-pathogens related to poultry industry. Both PEG and EOs led to the formation of flexible PLA/PEG/EO films with significant drop in the glass transition temperature (Tg ), and mechanical property. Time-temperature superposition (TTS) principle was employed to melt rheology of EO-based films at selected temperature, and rheological moduli superimposed well in an extended frequency range. Among EOs, cinnamon and clove oil-based films (PLA/PEG/CIN and PLA/PEG/CLO) exhibited a complete zone of inhibition against C. jejuni at the maximum concentration (1.6 mL per 2 g PLA/PEG blend) whereas the garlic oil-based film (PLA/PEG/GAR) had the lowest activity.

  15. The impact of the LuxS mutation on phenotypic expression of factors critical for Campylobacter jejuni colonization.

    Science.gov (United States)

    Mou, Kathy T; Plummer, Paul J

    2016-08-30

    Studies have collectively shown the wide impact that luxS mutation has on the expression and function of various aspects of Campylobacter jejuni virulence. Previous work from our group demonstrated that LuxS mutagenesis negatively impacts colonization of the gastrointestinal tract of several host species. To determine what is responsible for the colonization defect, we used a mechanistic approach to understand how the luxS mutation affects the expression of key physiologic factors important to the colonization ability of C. jejuni. This included expression of genes from the CmeABC efflux system, cell morphology, and motility through mucin substrate between wildtype, luxS mutant, and luxS complement of the C. jejuni strains 11168 and/or IA3902. We also measured and compared the activated methyl cycle (AMC) metabolite levels of the IA3902 luxS mutant to wildtype. Results showed that mutagenesis of the luxS gene completely disrupted the AMC with altered concentrations of AMC metabolites both upstream and downstream of LuxS. Multidrug efflux pump genes cmeABC and cmeR showed no significant changes in expression levels within the luxS mutant. Though motility through mucin was not completely unaffected by the luxS mutation, the lack of differences in cell morphology between wildtype and luxS mutant suggest that morphology is not responsible for the slight changes in mucin penetration observed in one of our luxS mutants. Though additional studies are warranted, these findings suggest that the CmeABC multi-drug efflux pump, cell morphology and mucin penetration are not major mechanisms responsible for the luxS mutant's colonization defect in its host. PMID:27527763

  16. Transducer Like Proteins of Campylobacter jejuni 81-176: Role in chemotaxis and colonization of the chicken gastrointestinal tract

    Directory of Open Access Journals (Sweden)

    Gireesh eRajashekara

    2015-05-01

    Full Text Available Transducer Like Proteins (Tlps, also known as Methyl accepting chemotaxis proteins (MCP, enable enteric pathogens to respond to changing nutrient levels in the environment by mediating taxis towards or away from specific chemoeffector molecules such as nutrients. Despite recent advances in the characterization of chemotaxis responses in Campylobacter jejuni, the impact of Tlps on the adaptation of this pathogen to disparate niches and hosts is not fully characterized. The latter is particularly evident in the case of C. jejuni 81-176, a strain that is known to be highly invasive. Furthermore, the cytoplasmic group C Tlps (Tlp5, 6, and 8 was not extensively evaluated. Here, we investigated the role of C. jejuni 81-176 Tlps in chemotaxis towards various substrates, biofilm formation, in vitro interaction with human intestinal cells, and chicken colonization. We found that the ∆tlp6 and ∆tlp10 mutants exhibited decreased chemotaxis towards aspartate whereas the ∆tlp6 mutant displayed a decreased chemotaxis towards Tri-Carboxylic Acid (TCA cycle intermediates such as pyruvate, isocitrate, and succinate. Our findings also corroborated that more than one Tlp is involved in mediating chemotaxis towards the same nutrient. The deletion of tlps affected important phenotypes such as motility, biofilm formation, and invasion of human intestinal epithelial cells (INT-407. The ∆tlp8 mutant displayed increased motility in soft agar and showed decreased biofilm formation. The ∆tlp8 and ∆tlp9 mutants were significantly defective in invasion in INT-407 cells. The ∆tlp10 mutant was defective in colonization of the chicken proximal and distal gastrointestinal tract, while the ∆tlp6 and ∆tlp8 mutants showed reduced colonization of the duodenum and jejunum. Our results highlight the importance of Tlps in C. jejuni’s adaptation and pathobiology.

  17. Bacteria in an intense competition for iron: Key component of the Campylobacter jejuni iron uptake system scavenges enterobactin hydrolysis product.

    Science.gov (United States)

    Raines, Daniel J; Moroz, Olga V; Blagova, Elena V; Turkenburg, Johan P; Wilson, Keith S; Duhme-Klair, Anne-K

    2016-05-24

    To acquire essential Fe(III), bacteria produce and secrete siderophores with high affinity and selectivity for Fe(III) to mediate its uptake into the cell. Here, we show that the periplasmic binding protein CeuE of Campylobacter jejuni, which was previously thought to bind the Fe(III) complex of the hexadentate siderophore enterobactin (Kd ∼ 0.4 ± 0.1 µM), preferentially binds the Fe(III) complex of the tetradentate enterobactin hydrolysis product bis(2,3-dihydroxybenzoyl-l-Ser) (H5-bisDHBS) (Kd = 10.1 ± 3.8 nM). The protein selects Λ-configured [Fe(bisDHBS)](2-) from a pool of diastereomeric Fe(III)-bisDHBS species that includes complexes with metal-to-ligand ratios of 1:1 and 2:3. Cocrystal structures show that, in addition to electrostatic interactions and hydrogen bonding, [Fe(bisDHBS)](2-) binds through coordination of His227 and Tyr288 to the iron center. Similar binding is observed for the Fe(III) complex of the bidentate hydrolysis product 2,3-dihydroxybenzoyl-l-Ser, [Fe(monoDHBS)2](3-) The mutation of His227 and Tyr288 to noncoordinating residues (H227L/Y288F) resulted in a substantial loss of affinity for [Fe(bisDHBS)](2-) (Kd ∼ 0.5 ± 0.2 µM). These results suggest a previously unidentified role for CeuE within the Fe(III) uptake system of C. jejuni, provide a molecular-level understanding of the underlying binding pocket adaptations, and rationalize reports on the use of enterobactin hydrolysis products by C. jejuni, Vibrio cholerae, and other bacteria with homologous periplasmic binding proteins.

  18. Bacteria in an intense competition for iron: Key component of the Campylobacter jejuni iron uptake system scavenges enterobactin hydrolysis product

    Science.gov (United States)

    Raines, Daniel J.; Moroz, Olga V.; Blagova, Elena V.; Turkenburg, Johan P.; Wilson, Keith S.

    2016-01-01

    To acquire essential Fe(III), bacteria produce and secrete siderophores with high affinity and selectivity for Fe(III) to mediate its uptake into the cell. Here, we show that the periplasmic binding protein CeuE of Campylobacter jejuni, which was previously thought to bind the Fe(III) complex of the hexadentate siderophore enterobactin (Kd ∼ 0.4 ± 0.1 µM), preferentially binds the Fe(III) complex of the tetradentate enterobactin hydrolysis product bis(2,3-dihydroxybenzoyl-l-Ser) (H5-bisDHBS) (Kd = 10.1 ± 3.8 nM). The protein selects Λ-configured [Fe(bisDHBS)]2− from a pool of diastereomeric Fe(III)-bisDHBS species that includes complexes with metal-to-ligand ratios of 1:1 and 2:3. Cocrystal structures show that, in addition to electrostatic interactions and hydrogen bonding, [Fe(bisDHBS)]2− binds through coordination of His227 and Tyr288 to the iron center. Similar binding is observed for the Fe(III) complex of the bidentate hydrolysis product 2,3-dihydroxybenzoyl-l-Ser, [Fe(monoDHBS)2]3−. The mutation of His227 and Tyr288 to noncoordinating residues (H227L/Y288F) resulted in a substantial loss of affinity for [Fe(bisDHBS)]2− (Kd ∼ 0.5 ± 0.2 µM). These results suggest a previously unidentified role for CeuE within the Fe(III) uptake system of C. jejuni, provide a molecular-level understanding of the underlying binding pocket adaptations, and rationalize reports on the use of enterobactin hydrolysis products by C. jejuni, Vibrio cholerae, and other bacteria with homologous periplasmic binding proteins. PMID:27162326

  19. AI-2 does not function as a quorum sensing molecule in Campylobacter jejuni during exponential growth in vitro

    Directory of Open Access Journals (Sweden)

    Winzer Klaus

    2009-10-01

    Full Text Available Abstract Background Campylobacter jejuni contains a homologue of the luxS gene shown to be responsible for the production of the signalling molecule autoinducer-2 (AI-2 in Vibrio harveyi and Vibrio cholerae. The aim of this study was to determine whether AI-2 acted as a diffusible quorum sensing signal controlling C. jejuni gene expression when it is produced at high levels during mid exponential growth phase. Results AI-2 activity was produced by the parental strain NCTC 11168 when grown in rich Mueller-Hinton broth (MHB as expected, but interestingly was not present in defined Modified Eagles Medium (MEM-α. Consistent with previous studies, the luxS mutant showed comparable growth rates to the parental strain and exhibited decreased motility halos in both MEM-α and MHB. Microarray analysis of genes differentially expressed in wild type and luxS mutant strains showed that many effects on mRNA transcript abundance were dependent on the growth medium and linked to metabolic functions including methionine metabolism. Addition of exogenously produced AI-2 to the wild type and the luxS mutant, growing exponentially in either MHB or MEM-α did not induce any transcriptional changes as analysed by microarray. Conclusion Taken together these results led us to conclude that there is no evidence for the role of AI-2 in cell-to-cell communication in C. jejuni strain NCTC 11168 under the growth conditions used, and that the effects of the luxS mutation on the transcriptome are related to the consequential loss of function in the activated methyl cycle.

  20. Effect of refrigerated and frozen storage on the survival of Campylobacter jejuni in cooked chicken meat breast.

    Science.gov (United States)

    Eideh, Ala'a M F; Al-Qadiri, Hamzah M

    2011-01-01

    This experimental work aimed to examine the survivability of Campylobacter jejuni in cooked chicken breast under several conditions: storage for 1, 3, and 7 d at refrigerated temperatures (4 °C) and for 20 d at frozen temperatures (-18 °C). In addition, storage at ambient temperature (26 to 28 °C) was involved. Chicken samples were inoculated with a mixed culture of C. jejuni strains (ATCC: 29428 and 33219) of known concentrations (50 and 500 CFU/g). Bacterial cells were recovered and enumerated using standard procedure (Preston method). Bacteria were not detected in the majority of samples stored at ambient temperature. Refrigeration reduced survivals in 95, 90, and 77.5% for samples inoculated with 500 CFU/g and kept for 1, 3, and 7 d, respectively. The maximum reduction reached 1 log(10) cycle for all refrigeration durations. It was observed that bacteria died in 17.5% of samples kept for 7 d at 4 °C. However, survivors in samples inoculated with 50 CFU/g were not detected in 50, 65, and 55% of samples kept for 1, 3, and 7 d, respectively. Freezing rendered survivors not detectable in 70% of samples inoculated with 50 CFU/g, while survived viable counts were reduced in 92.5% of samples inoculated with 500 CFU/g. These findings suggested that C. jejuni could be killed or just sublethally injured with or without reduction in viable counts under the investigated storage temperatures, which may indicate the ability of this bacterium to survive in chicken meat stored under refrigerated and frozen conditions. PMID:21535688

  1. A fluoroquinolone resistance associated mutation in gyrA affects DNA supercoiling in Campylobacter jejuni

    Directory of Open Access Journals (Sweden)

    Jing eHan

    2012-03-01

    Full Text Available The prevalence of fluoroquinolone (FQ-resistant Campylobacter has become a concern for public health. To facilitate the control of FQ-resistant Campylobacter, it is necessary to understand the impact of FQ resistance on the fitness of Campylobacter in its natural hosts as understanding fitness will help to determine and predict the persistence of FQ-resistant Campylobacter. Previously it was shown that acquisition of resistance to FQ antimicrobials enhanced the in vivo fitness of FQ-resistant Campylobacter. In this study, we confirmed the role of the Thr-86-Ile mutation in GyrA in modulating Campylobacter fitness by reverting the mutation to the wild-type allele, which resulted in the loss of the fitness advantage. Additionally, we determined if the resistance-conferring GyrA mutations alter the enzymatic function of the DNA gyrase. Recombinant wild-type gyrase and mutant gyrases with three different types of mutations (Thr-86-Ile, Thr-86-Lys, and Asp-90-Asn, which are associated with FQ resistance in Campylobacter, were generated in E. coli and compared for their supercoiling activities using an in vitro assay. The mutant gyrase with the Thr-86-Ile change showed a greatly reduced supercoiling activity compared with the wild-type gyrase, while other mutant gyrases did not show an altered supercoiling. Furthermore, we measured DNA supercoiling within Campylobacter cells using a reporter plasmid. Consistent with the results from the in vitro supercoiling assay, the FQ-resistant mutant carrying the Thr-86-Ile change in GyrA showed much less DNA supercoiling than the wild-type strain and the mutant strains carrying other mutations. Together, these results indicate that the Thr-86-Ile mutation, which is predominant in clinical FQ-resistant Campylobacter, modulates DNA supercoiling homeostasis in FQ-resistant Campylobacter.

  2. THERMOTOLERANT Campylobacter SPECIES ISOLATED FROM PSITTACIFORMES IN THE PERUVIAN AMAZON REGION

    Directory of Open Access Journals (Sweden)

    Alvaro TRESIERRA-AYALA

    1998-07-01

    Full Text Available Foi determinada a freqüência de isolamento de campylobacters termotolerantes em Psittaciformes silvestres capturados na região amazônica do Peru. Campylobacters foram isolados em 10/142 (7.0% dos animais estudados, sendo C. jejuni subsp. jejuni biovar I (6/10 o mais freqüente, seguido de C. coli biovar II (2/10, C. lari não foi isolado. Os resultados sugerem que estas aves podem ser importantes reservatórios destas bactérias.

  3. Intracellular pH as an indicator of viability and resuscitation of Campylobacter jejuni after decontamination with lactic acid

    DEFF Research Database (Denmark)

    Smigic, Nada; Rajkovic, Andreja; Nielsen, Dennis Sandris;

    2009-01-01

    The aim of the study was to determine intracellular pH (pH(i)) as an indicator of the physiological state of two Campylobacter jejuni strains (603 and 608) at the single cell level after bactericidal treatment with lactic acid (3% v/v lactic acid, pH 4.0, 0.85% w/v NaCl) and during recovery and s...

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

    Directory of Open Access Journals (Sweden)

    Hana eTuronova

    2015-07-01

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

  5. Signal balancing by the CetABC and CetZ chemoreceptors controls energy taxis in Campylobacter jejuni.

    Directory of Open Access Journals (Sweden)

    Mark Reuter

    Full Text Available The coupling of environmental sensing to flagella-mediated directed motility allows bacteria to move to optimum environments for growth and survival, either by sensing external stimuli (chemotaxis or monitoring internal metabolic status (energy taxis. Sensing is mediated by transducer-like proteins (Tlp, either located in the membrane or in the cytoplasm, which commonly influence motility via the CheA-CheY chemotaxis pathway. In this study we have investigated the role of PAS-domain-containing intracellular Tlp-sensors in energy taxis of the food-borne pathogen Campylobacter jejuni, using plate- and tube-based assays utilising the conversion of the redox indicator dyes triphenyl tetrazolium chloride (TTC and resazurin. Inactivation of the genes encoding the Campylobacter Energy Taxis system (CetA (Tlp9 and CetB (Aer2 in C. jejuni strain NCTC 11168 resulted in reduced taxis. Inactivation of the cj1191c gene, encoding the CetB homolog CetC (Aer1, did not affect taxis per se, but the cetC gene complemented a cetB mutant in trans, indicating that CetC can form a functional signal transduction complex with CetA in the absence of CetB. Inactivation of both CetB and CetC resulted in greatly reduced taxis confirming the role of CetC in energy taxis. Inactivation of the cj1110c gene, encoding Tlp8 (CetZ, a cytoplasmic sensor with two PAS-domains, resulted in increased taxis, a phenotype opposite to that of CetAB. Inactivation of the cheA gene resulted in the same overall phenotype as the cetAB mutant in both wild-type and cetZ backgrounds, suggesting that both systems use the CheA system for signal transduction. Absence of both CetAB and CetZ resulted in the cetAB taxis phenotype, suggesting that CetZ is subordinate to CetAB. In conclusion, we present evidence that C. jejuni balances the input from two counteracting PAS-domain-containing sensory systems to position itself for optimal usage of energy resources.

  6. Effect of bacteriophage application on Campylobacter jejuni loads in commercial broiler flocks.

    Science.gov (United States)

    Kittler, Sophie; Fischer, Samuel; Abdulmawjood, Amir; Glünder, Gerhard; Klein, Günter

    2013-12-01

    Campylobacteriosis is the most frequent food-borne human enteritis. The major source for infection with Campylobacter spp. is broiler meat. Risk assessments consider the reduction of Campylobacter in primary production to be most beneficial for human health. The aim of this study was to test the efficacy of a bacteriophage application under commercial conditions which had proved to be effective in previous noncommercial studies under controlled experimental conditions. A phage cocktail for Campylobacter reduction was tested on three commercial broiler farms each with a control and an experimental group. Colonization of Campylobacter was confirmed prior to phage application in fecal samples. Subsequently, a phage cocktail was applied via drinking water in the experimental group (log10 5.8 to 7.5 PFU/bird). One day after phage application, Campylobacter counts of one experimental group were reduced under the detection limit (log10 3.2 CFU/g cecal content compared to the control was still detected (P=0.0011). No significant reduction was observed in the experimental groups of the other trials. However, a significant drop in cecal Campylobacter counts occurred in a phage-contaminated control. These results suggest that maximum reduction of Campylobacter at the slaughterhouse might be achieved by phage application 1 to 4 days prior to slaughter.

  7. Campylobacter jejuni acquire new host-derived CRISPR spacers when in association with bacteriophages harbouring a CRISPR-like Cas4 protein

    Directory of Open Access Journals (Sweden)

    Ian F. Connerton

    2015-01-01

    Full Text Available Campylobacter jejuni is a worldwide cause of human diarrhoeal disease. Clustered Repetitively Interspaced Palindromic Repeats (CRISPRs and associated proteins allow Bacteria and Archaea to evade bacteriophage and plasmid infection. Type II CRISPR systems are found in association with combinations of genes encoding the CRISPR-associated Cas1, Cas2, Cas4 or Csn2, and Cas9 proteins. C. jejuni possesses a minimal subtype II-C CRISPR system containing cas1, cas2, and cas9 genes whilst cas4 is notably absent. Cas4 proteins possess 5ʹ-3ʹ exonuclease activity to create recombinogenic-ends for spacer acquisition. Here we report a conserved Cas4-like protein in Campylobacter bacteriophages that creates a novel split arrangement between the bacteriophage and host that represents a new twist in the bacteriophage/host co-evolutionary arms race. The continuous association of bacteriophage and host in the carrier state life cycle of C. jejuni provided an opportunity to study spacer acquisition in this species. Remarkably all the spacer sequences observed were of host origin. We hypothesise that Campylobacter bacteriophages can use Cas4-like protein to activate spacer acquisition to use host DNA as an effective decoy to bacteriophage DNA. Bacteria that acquire self-spacers and escape phage infection must overcome CRISPR-mediated autoimmunity either by loss of the interference functions leaving them susceptible to foreign DNA incursion or tolerate changes in gene regulation.

  8. Enumeração de Campylobacter spp. e presença de Campylobacter jejuni em carcaças de frango no estado de Minas Gerais Enumeration of Campylobacter spp. and presence of Campylobacter jejuni in broiler carcasses in the state of Minas Gerais, Brazil

    Directory of Open Access Journals (Sweden)

    Afonso de Liguori Oliveira

    2013-03-01

    Full Text Available Campilobacteriose é uma zoonose de distribuição mundial, com repercussões importantes na saúde pública e um grande impacto socioeconômico. As rotas de transmissão ao homem das espécies de Campylobacter são o contato direto com animais portadores, água ou alimentos contaminados e é mais comumente associado ao consumo de carne de frango. O objetivo deste estudo foi detectar e enumerar Campylobacter spp. por um método rápido (SimPlate® e pelo método convencional (plaqueamento direto em ágar mCCD, determinando a frequência e o nível de contaminação em 75 amostras de carcaças de frangos (25 Pré-chiller e 25 Pós-chiller obtidos em abatedouros sob inspeção federal no Estado de Minas Gerais e 25 amostras congeladas, coletadas no comércio verejista. As amostras avaliadas, independentemente do método utilizado, apresentaram Campylobacter spp. em 56,0% das carcaças antes do chiller e 44,0% após o chiller. A contagem das amostras congeladas se apresentou abaixo do limite de detecção do método. Das 75 amostras testadas, 34,7% foram positivas para Campylobacter spp. Das amostras positivas, 38,5% foram identificadas como C. jejuni e, entre estas, o maior número de C. jejuni foi obtido nas carcaças de frango antes do chiller (42,9%, seguido de carcaça de frango após o chiller (36,4%. O estudo mostrou que a detecção e enumeração é variável, embora em nenhuma das amostras destinadas ao comércio foram obtidas contagens acima da dose infectante estabelecida. O uso do SimPlate® foi adequado e equivalente ao plaqueamento direto em ágar mCCD e apresentou uma melhor taxa de recuperação de células de Campylobacter spp.Campylobacteriosis is a worldwide distribution zoonosis, with significant repercussions on public health and with a high socioeconomic impact. Routes of transmission of Campylobacter species to man are direct contact with carrier animals, contaminated water or food and it is most commonly associated with

  9. Bacteriophage F336 Recognizes the Capsular Phosphoramidate Modification of Campylobacter jejuni NCTC11168

    DEFF Research Database (Denmark)

    Sørensen, Martine C. Holst; van Alphen, Lieke B.; Harboe, Anne;

    2011-01-01

    indicating that the receptor is altered. While proteinase K-treated C. jejuni cells did not affect adsorption, periodate treatment resulted in reduced adsorption, suggesting that the phage binds to a carbohydrate moiety. Using high-resolution magic angle spinning nuclear magnetic resonance (NMR) spectroscopy...... source of human campylobacteriosis. In this study, we used phage F336 belonging to the Myoviridae family to select a C. jejuni NCTC11168 phage-resistant strain, called 11168R, with the aim of investigating the mechanisms of phage resistance. We found that phage F336 has reduced adsorption to 11168R, thus...

  10. Isolation of Campylobacter fetus from a pet turtle.

    OpenAIRE

    Harvey, S.; Greenwood, J. R.

    1985-01-01

    During the course of a Salmonella agona case investigation, Campylobacter fetus was isolated from a pet turtle. This is the first reported isolation of C. fetus from a turtle and suggests that turtles, in addition to being reservoirs for Salmonella species, may also be reservoirs for C. fetus.

  11. Detection and analysis of campylobacter jejuni in 412 diarrhea cases of Guiyang%贵阳市412例腹泻病例空肠弯曲菌的检测分析

    Institute of Scientific and Technical Information of China (English)

    韦小瑜; 游旅; 马青; 刘荚

    2011-01-01

    目的 建立常规的空肠弯曲菌检测方法,了解贵阳市腹泻人群空肠弯曲菌的感染状况.方法 采用简单生化及多重PCR方法 对贵阳市412例腹泻人群进行空肠弯曲菌检测.结果 412份腹泻标本中,7份空肠弯曲菌阳性,平均检出率1.70%.1岁以下婴幼儿感染率与其它人群差异无统计学意义(P>0.05).男女患者感染率差异无统计学意义(P>0.05).粘液便空肠弯曲菌检出率(7.69%)显著高于水样便(0.60%)(P<0.01).结论 利用简单生化及多重PER方法 能很好地检出空肠弯曲菌;贵阳市腹泻人群中存在空肠弯曲菌的感染,且主要感染婴幼儿,需加强婴幼儿腹泻中空肠弯曲菌感染的监测.%Objective To establish routine testing methods and understand campylobacter jejuni infection status in diarrhea patients in Guiyang. Method Using simple biochemical methods and multiple PCR methods to detect the campylobacter jejuni in 412 diarrhea patients in Guiyang city. Results 7 c jejuni strains were isolated from 412 diarrhea cases,and with the average detection rate of 1.70% . The infection rate of 1 year old had no significant difference with other crowds , the detection rate of men and female had no significant difference too. The detection rate of mucus stool (7. 69% ) was higher than watery stool (0. 60% ) ( P < 0. 01) . Conclusions Simple biochemical methods and multiple PCR methods were better for testing c jejuni. There were C jejuni infection in diarrhea patients in Guiyang city, and it mainly infected babies and infants, so we need to strengthen the momitoring of baby diarrhea

  12. Rapid paracellular transmigration of Campylobacter jejuni across polarized epithelial cells without affecting TER: role of proteolytic-active HtrA cleaving E-cadherin but not fibronectin

    LENUS (Irish Health Repository)

    Boehm, Manja

    2012-04-25

    AbstractBackgroundCampylobacter jejuni is one of the most important bacterial pathogens causing food-borne illness worldwide. Crossing the intestinal epithelial barrier and host cell entry by C. jejuni is considered the primary reason of damage to the intestinal tissue, but the molecular mechanisms as well as major bacterial and host cell factors involved in this process are still widely unclear.ResultsIn the present study, we characterized the serine protease HtrA (high-temperature requirement A) of C. jejuni as a secreted virulence factor with important proteolytic functions. Infection studies and in vitro cleavage assays showed that C. jejuni’s HtrA triggers shedding of the extracellular E-cadherin NTF domain (90 kDa) of non-polarised INT-407 and polarized MKN-28 epithelial cells, but fibronectin was not cleaved as seen for H. pylori’s HtrA. Deletion of the htrA gene in C. jejuni or expression of a protease-deficient S197A point mutant did not lead to loss of flagella or reduced bacterial motility, but led to severe defects in E-cadherin cleavage and transmigration of the bacteria across polarized MKN-28 cell layers. Unlike other highly invasive pathogens, transmigration across polarized cells by wild-type C. jejuni is highly efficient and is achieved within a few minutes of infection. Interestingly, E-cadherin cleavage by C. jejuni occurs in a limited fashion and transmigration required the intact flagella as well as HtrA protease activity, but does not reduce transepithelial electrical resistance (TER) as seen with Salmonella, Shigella, Listeria or Neisseria.ConclusionThese results suggest that HtrA-mediated E-cadherin cleavage is involved in rapid crossing of the epithelial barrier by C. jejuni via a very specific mechanism using the paracellular route to reach basolateral surfaces, but does not cleave the fibronectin receptor which is necessary for cell entry.

  13. Rapid paracellular transmigration of Campylobacter jejuni across polarized epithelial cells without affecting TER: role of proteolytic-active HtrA cleaving E-cadherin but not fibronectin

    Directory of Open Access Journals (Sweden)

    Boehm Manja

    2012-04-01

    Full Text Available Abstract Background Campylobacter jejuni is one of the most important bacterial pathogens causing food-borne illness worldwide. Crossing the intestinal epithelial barrier and host cell entry by C. jejuni is considered the primary reason of damage to the intestinal tissue, but the molecular mechanisms as well as major bacterial and host cell factors involved in this process are still widely unclear. Results In the present study, we characterized the serine protease HtrA (high-temperature requirement A of C. jejuni as a secreted virulence factor with important proteolytic functions. Infection studies and in vitro cleavage assays showed that C. jejuni’s HtrA triggers shedding of the extracellular E-cadherin NTF domain (90 kDa of non-polarised INT-407 and polarized MKN-28 epithelial cells, but fibronectin was not cleaved as seen for H. pylori’s HtrA. Deletion of the htrA gene in C. jejuni or expression of a protease-deficient S197A point mutant did not lead to loss of flagella or reduced bacterial motility, but led to severe defects in E-cadherin cleavage and transmigration of the bacteria across polarized MKN-28 cell layers. Unlike other highly invasive pathogens, transmigration across polarized cells by wild-type C. jejuni is highly efficient and is achieved within a few minutes of infection. Interestingly, E-cadherin cleavage by C. jejuni occurs in a limited fashion and transmigration required the intact flagella as well as HtrA protease activity, but does not reduce transepithelial electrical resistance (TER as seen with Salmonella, Shigella, Listeria or Neisseria. Conclusion These results suggest that HtrA-mediated E-cadherin cleavage is involved in rapid crossing of the epithelial barrier by C. jejuni via a very specific mechanism using the paracellular route to reach basolateral surfaces, but does not cleave the fibronectin receptor which is necessary for cell entry.

  14. Comparison of cstⅡ gene in Guillain-Barré syndrome-associated Campylobacter jejuni strains%吉兰-巴雷综合征相关空肠弯曲菌cstⅡ基因序列对比研究

    Institute of Scientific and Technical Information of China (English)

    孙世超; 白欣立; 陈娟; 王莹; 邢丛丛; 付金生; 李震中

    2011-01-01

    目的 研究华北地区3株吉兰-巴雷综合征(Guillain-Barré syndrome,GBS)相关空肠弯曲菌(Campylobacter jejuni,C.jejuni)的cstⅡ基因序列并寻找GBS相关及GBS无关的两类空肠弯曲菌菌株间可能与GBS致病性相关的基因突变位点和蛋白高级结构变化,分析菌株间遗传进化关系.方法 选取分离自GBS患者粪便并经动物模型证实为致GBS的3株空肠弯曲菌菌株进行培养并提取基因组DNA测序,与GenBank中GBS无关空肠弯曲菌基因序列进行配对对比,寻找两类菌株间可能导致空肠弯曲菌致病能力差异的cstⅡ基因碱基突变位点及蛋白高级结构改变,观察这种变化是否为GBS相关菌株的共性,并构建遗传进化树.结果 与GBS无关菌株相比,3株致GBS菌株cstⅡ基因的氨基酸序列二级结构的第7个α螺旋的165~180区段均被打开形成了折叠或转角,该变化在其他GBS相关菌株的二级结构变化中同样得到体现.实验中75%的GBS相关菌株cstⅡ基因为双功能,25%GBS相关菌株及所有GBS无关菌株cstⅡ基因为单功能.LL株与实验中涉及的GBS相关菌株在进化上高度类聚.结论 双功能cstⅡ可能与空肠弯曲菌的致GBS特性相关.LL株cstⅡ基因在一定程度上反映了亚洲地区的GBS相关菌株cstⅡ基因的序列特征,为进一步探索GBS发病的地域性特点并对GBS菌株进行监测提供了资料.%Objective To investigate the pathogenic mechanism of Campylobacter jejuni(C.jejuni) associated with Guillain-Barré syndrome(GBS) and provide strategy for gene modification, the cstⅡ gene from 8 GBS-associated C.jejuni strains were compared with that from 3 GBS-unrelated C.jejuni strains, getting the base and amino acid mutations, the changes of secondary structures and finding the region which may be responsible for the pathogenicity of C.jejuni inducing GBS. Methods Three GBS-associated C.jejuni strains isolated from stools of GBS patients in north China were

  15. Complete Genome Sequence of Campylobacter iguaniorum Strain RM11343, Isolated from an Alpaca.

    Science.gov (United States)

    Miller, William G; Yee, Emma; Huynh, Stephen; Chapman, Mary H; Parker, Craig T

    2016-01-01

    Campylobacter iguaniorum is a member of the C. fetus group of campylobacters and is one of two Campylobacter taxa isolated from reptiles. This study describes the whole-genome sequence of the C. iguaniorum strain RM11343, which was isolated from a California alpaca fecal sample. PMID:27365359

  16. Structural analysis of PseH, the Campylobacter jejuni N-acetyltransferase involved in bacterial O-linked glycosylation

    Energy Technology Data Exchange (ETDEWEB)

    Song, Wan Seok; Nam, Mi Sun; Namgung, Byeol [Department of Systems Immunology, College of Biomedical Science, Kangwon National University, Chuncheon 200-701 (Korea, Republic of); Yoon, Sung-il, E-mail: sungil@kangwon.ac.kr [Department of Systems Immunology, College of Biomedical Science, Kangwon National University, Chuncheon 200-701 (Korea, Republic of); Institute of Bioscience and Biotechnology, Kangwon National University, Chuncheon 200-701 (Korea, Republic of)

    2015-03-20

    Campylobacter jejuni is a bacterium that uses flagella for motility and causes worldwide acute gastroenteritis in humans. The C. jejuni N-acetyltransferase PseH (cjPseH) is responsible for the third step in flagellin O-linked glycosylation and plays a key role in flagellar formation and motility. cjPseH transfers an acetyl group from an acetyl donor, acetyl coenzyme A (AcCoA), to the amino group of UDP-4-amino-4,6-dideoxy-N-acetyl-β-L-altrosamine to produce UDP-2,4-diacetamido-2,4,6-trideoxy-β-L-altropyranose. To elucidate the catalytic mechanism of cjPseH, crystal structures of cjPseH alone and in complex with AcCoA were determined at 1.95 Å resolution. cjPseH folds into a single-domain structure of a central β-sheet decorated by four α-helices with two continuously connected grooves. A deep groove (groove-A) accommodates the AcCoA molecule. Interestingly, the acetyl end of AcCoA points toward an open space in a neighboring shallow groove (groove-S), which is occupied by extra electron density that potentially serves as a pseudosubstrate, suggesting that the groove-S may provide a substrate-binding site. Structure-based comparative analysis suggests that cjPseH utilizes a unique catalytic mechanism of acetylation that has not been observed in other glycosylation-associated acetyltransferases. Thus, our studies on cjPseH will provide valuable information for the design of new antibiotics to treat C. jejuni-induced gastroenteritis. - Highlights: • cjPseH adopts a single-domain structure of a central β-sheet decorated by α-helices. • cjPseH features two continuously connected grooves on the protein surface. • Acetyl coenzyme A (AcCoA) binds into a deep groove of cjPseH in an ‘L’ shape. • The acetyl end of AcCoA points to a wide groove, a potential substrate-binding site.

  17. Complete genome sequence of Campylobacter iguaniorum strain 1485ET, isolated from a bearded dragon (Pogona vitticeps)

    Science.gov (United States)

    Campylobacter iguaniorum has been isolated from reptiles. This Campylobacter species is genetically related to C. fetus and C. hyointestinalis. Here we present the first whole genome sequence for this species....

  18. Campylobacter jejuni strains of human and chicken origin are invasive in chickens after oral challenge

    DEFF Research Database (Denmark)

    Knudsen, Katrine Nørrelund; Bang, Dang Duong; Andresen, Lars Ole;

    2006-01-01

    -old chickens hatched from specific pathogen free (SPF) eggs. Of the four C jejuni strains tested, three were Penner heat-stable serotype 2,flaA type 1/1, the most common type found among broilers and human cases in Denmark. The fourth strain was Penner heat-stable serotype 19, which has been shown...... to be associated with the Guillain Barre Syndrome (GBS) in humans. The minimum dose for establishing colonization in the clay-old chickens was approximately 2 cfu, whereas two- to threefold higher doses were required for establishing colonization in the 14-day-old chickens. Two of the C jejuni strains were shown...... to be invasive in orally challenged chickens as well as in three different human epithelial cell lines....

  19. Immunogenicety and immunoprotection of recombinant PEB1 in Campylobacter-jejuni-infected mice

    Institute of Scientific and Technical Information of China (English)

    Lian-Feng Du; Zhen-Jiang Li; Xian-Ying Tang; Jun-Qiong Huang; Wan-Bang Sun

    2008-01-01

    AIM: To construct a prokaryotic expression vector carrying Carnpylobacter jejuni peb1A gene and express it in Escherichia coli. Immunoreactivity and antigenicity of rPEB1 were evaluated. The ability of rPEB1 to induce antibody responses and protective efficacy was identified. METHODS: peb1A gene was amplified by PCR, target gene and prokaryotic expression plasmid pET28a (+) was digested with BamHI and XhoI, respectively. DNA was ligated with T4 DNA ligase to construct recombinant plasmid pET28a(+)-peb1A. The rPEB1 was expressed in E. coli BL21 (DE3) and identified by SDS-PAGE. BALB/c mice were immunized with rPEB1. ELISA was used to detect the specific antibody titer and MTT method was used to measure the stimulation index of spleen lymphocyte transformation. RESULTS: The recombinant plasmid pET28a (+)-peb1A was correctly constructed. The expression output of PEB1 protein in pET28a (+)-peb1A system was approximately 33% of total proteins in E.coli The specific IgG antibody was detected in serum of BALB/c mice immunized with rPEB1 protein. Effective immunological protection with a lower sicknessincidence and mortality was seen in the mice suffering from massive C. jejuni infection. CONCLUSION: rPEB1 protein is a valuable candidate for C. jejuni subunit vaccine.

  20. Analysis of the LIV System of Campylobacter jejuni Reveals Alternative Roles for LivJ and LivK in Commensalism beyond Branched-Chain Amino Acid Transport ▿

    OpenAIRE

    Ribardo, Deborah A.; Hendrixson, David R.

    2011-01-01

    Campylobacter jejuni is a leading cause of diarrheal disease in humans and an intestinal commensal in poultry and other agriculturally important animals. These zoonotic infections result in significant amounts of C. jejuni present in the food supply to contribute to disease in humans. We previously found that a transposon insertion in Cjj81176_1038, encoding a homolog of the Escherichia coli LivJ periplasmic binding protein of the leucine, isoleucine, and valine (LIV) branched-chain amino aci...

  1. Campylobacter jejuni adenosine triphosphate phosphoribosyltransferase is an active hexamer that is allosterically controlled by the twisting of a regulatory tail.

    Science.gov (United States)

    Mittelstädt, Gerd; Moggré, Gert-Jan; Panjikar, Santosh; Nazmi, Ali Reza; Parker, Emily J

    2016-08-01

    Adenosine triphosphate phosphoribosyltransferase (ATP-PRT) catalyzes the first committed step of the histidine biosynthesis in plants and microorganisms. Here, we present the functional and structural characterization of the ATP-PRT from the pathogenic ε-proteobacteria Campylobacter jejuni (CjeATP-PRT). This enzyme is a member of the long form (HisGL ) ATP-PRT and is allosterically inhibited by histidine, which binds to a remote regulatory domain, and competitively inhibited by AMP. In the crystalline form, CjeATP-PRT was found to adopt two distinctly different hexameric conformations, with an open homohexameric structure observed in the presence of substrate ATP, and a more compact closed form present when inhibitor histidine is bound. CjeATP-PRT was observed to adopt only a hexameric quaternary structure in solution, contradicting previous hypotheses favoring an allosteric mechanism driven by an oligomer equilibrium. Instead, this study supports the conclusion that the ATP-PRT long form hexamer is the active species; the tightening of this structure in response to remote histidine binding results in an inhibited enzyme. PMID:27191057

  2. Guillain-Barré syndrome and Campylobacter jejuni infection%GBS与空肠弯曲菌感染

    Institute of Scientific and Technical Information of China (English)

    吴铮; 范文辉

    2005-01-01

    吉兰-巴雷综合征(Guillain-Barré syndrome,GBS)表现为一种由前驱细菌或病毒感染所触发的自限性自身免疫疾病,其中空肠弯曲菌(Campylobacter jejuni,CJ)是最常见的前驱感染病原,研究提示血清型O(Penner)O:1,O:2,O:4,O:10,O:19,O:23,O:36,O:41等CJ菌株的脂多糖(lipopolysaccharide,LPS)在分子结构上与人类神经节苷脂表位之间具有分子模拟现象,从而导致机体对二者的交叉反应产生抗-神经节苷脂自身抗体可能是GBS的发病机理.抗-神经节苷脂抗体在不同类型的GBS患者中具有一定的特异性,这个特点在急性周围神经病中可用于临床诊断和估计预后.

  3. Characterization of CetA and CetB, a bipartite energy taxis system in Campylobacter jejuni.

    Science.gov (United States)

    Elliott, Kathryn T; Dirita, Victor J

    2008-09-01

    The energy taxis receptor Aer, in Escherichia coli, senses changes in the redox state of the electron transport system via an flavin adenine dinucleotide cofactor bound to a PAS domain. The PAS domain (a sensory domain named after three proteins Per, ARNT and Sim, where it was first identified) is thought to interact directly with the Aer HAMP domain to transmit this signal to the highly conserved domain (HCD) found in chemotaxis receptors. An apparent energy taxis system in Campylobacter jejuni is composed of two proteins, CetA and CetB, that have the domains of Aer divided between them. CetB has a PAS domain, while CetA has a predicted transmembrane region, HAMP domain and the HCD. In this study, we examined the expression of cetA and cetB and the biochemical properties of the proteins they encode. cetA and cetB are co-transcribed independently of the flagellar regulon. CetA has two transmembrane helices in a helical hairpin while CetB is a peripheral membrane protein tightly associated with the membrane. CetB levels are CetA dependent. Additionally, we demonstrated that both CetA and CetB participate in complexes, including a likely CetB dimer and a complex that may include both CetA and CetB. This study provides a foundation for further characterization of signal transduction mechanisms within CetA/CetB. PMID:18631239

  4. Detection of Campylobacter species using monoclonal antibodies

    Science.gov (United States)

    Young, Colin R.; Lee, Alice; Stanker, Larry H.

    1999-01-01

    A panel of species specific monoclonal antibodies were raised to Campylobacter coli, Campylobacter jejuni and Campylobacter lari. The isotypes, and cross-reactivity profiles of each monoclonal antibody against an extensive panel of micro- organisms, were determined.

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

    Science.gov (United States)

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

    2016-01-01

    Two independent trials were carried out to evaluate the effect of feed form, whole wheat (WW) and oat hulls (OH) addition on gastrointestinal (GIT) weight and Campylobacter jejuni colonization in orally infected birds. In Trial 1, there were six treatments factorially arranged with two feed forms (mash vs pellets), and three levels of WW from 1-21/22-42d: 0/0, 7.5/15%, 15/30%. Broilers were allocated in cages (3 birds/cage, 12 cages/treatment). In Trial 2, there were three treatments: a mash diet, a mash diet including WW (7.5% from 1–21 and 15% from 22-42d), and a third treatment including also 5%OH. Broilers were allocated in floor pens (1 pen with 30 birds/treatment). At 14d, all broilers in Trial 1 or 3 broilers/pen in Trial 2 were orally challenged with 1.5 x 105 cfu of C. jejuni ST-45 /. In Trial 1, birds fed pelleted diets consumed 13.5% more feed, gained 31% more weight, and presented 12.9% better feed conversion for the whole trial (PGIT and gizzard and increased the relative weight of proventriculus (PGIT derived of FF and WW inclusion, no clear reductions in C. jejuni populations in the ceca were observed. However, WW and OH inclusion to mash diets significantly reduced cecal C. jejuni colonization at 42 days. PMID:27500730

  6. The Campylobacter jejuni RacRS two-component system activates the glutamate synthesis by directly upregulating γ-glutamyltranspeptidase (GGT

    Directory of Open Access Journals (Sweden)

    Anne-Xander evan der Stel

    2015-06-01

    Full Text Available The highly conserved enzyme γ-glutamyltranspeptidase (GGT plays an important role in metabolism of glutathione and glutamine. Yet, the regulation of ggt transcription in prokaryotes is poorly understood. In the human pathogen Campylobacter jejuni, GGT is important as it contributes to persistent colonization of the gut. Here we show that the GGT activity in C. jejuni is dependent on a functional RacRS (reduced ability to colonize two-component system. Electrophoretic mobility shift and luciferase reporter assays indicate that the response regulator RacR binds to a promoter region ~80 bp upstream of the ggt transcriptional start site, which contains a recently identified RacR DNA binding consensus sequence. RacR needs to be phosphorylated to activate the transcription of the ggt gene, which is the case under low oxygen conditions in presence of alternative electron acceptors. A functional GGT and RacR are needed to allow C. jejuni to grow optimally on glutamine as sole carbon source under RacR inducing conditions. However, when additional carbon sources are present C. jejuni is capable of utilizing glutamine independently of GGT. RacR is the first prokaryotic transcription factor known to directly upregulate both the cytoplasmic (glutamine-2-oxoglutarate aminotransferase (GOGAT as well as the periplasmic (GGT production of glutamate.

  7. Investigation and resistance analysis of Campylobacter jejuni in broiler%鸡源空肠弯曲菌调查与耐药性分析

    Institute of Scientific and Technical Information of China (English)

    孙桂霞; 徐世文

    2015-01-01

    Campylobacter jejuni is a kind of pathogenic bacteria which cause serious damage to the human and animal health. To understand the epidemiological and drug resistance of Campylobacter jejuni in broiler of Harbin, the present study investigated the infection status of Campylobacter jejuni in broiler in Harbin and its surrounding area, meanwhile, detected the drug resistance and expression of resistant genes. The results showed that the average infections of chicken farms were 35.67%. Campylobacter jejuni had different degrees of antibiotic resistance to 13 kinds of antibiotics, which the antibiotic resistance of tetracycline and ampicil in sodium were 100%, while there were multiple drug resistance. The highest drug-resistant spectrums were 11, while the detection rate of bacterial strain was 14.95%. 57.94%bacterial strain contained tetA, tetB, tetC, blaCTX-M, blaTEM and blaNDM-1 genes, that the detection rate were 57.94%, 66.36%, 28.97%, 42.99%, 55.14% and 8.41%, respectively. However, the present study didn't detect tetD and blaSHV-1 genes. The results showed that the infections of Campylobacter jejuni in chicken farms of Harbin were occured commonly, while there were multiple drug resistance. It provided the basis for effective prevention and treatment of Campylobacter jejuni.%空肠弯曲菌是严重危害人畜健康的病原菌,为了解哈尔滨市肉鸡源性空肠弯曲菌流行病学和耐药性,研究调查哈尔滨市周边肉鸡场空肠弯曲菌感染状况,通过耐药性和耐药基因检测,发现鸡场平均感染率为35.67%,对13种抗生素均有不同程度耐药性,四环素和氨苄西林钠达100%,存在多重耐药性,最高耐药谱达11个,检出菌株率为14.95%.57.94%菌株携带tetA、tetB、tetC、blaCTX-M、blaTEM和blaNDM-1基因,检出率分别为57.94%、66.36%、28.97%、42.99%、55.14%和8.41%,而tetD和blaSHV-1基因未检出.结果表明,哈尔滨市肉鸡场空肠弯曲菌感染普遍,存在多重耐药并携带多种耐药

  8. Rapid detection of Campylobacter jejuni by multiplex PCR technique%应用多重PCR快速检测空肠弯曲菌

    Institute of Scientific and Technical Information of China (English)

    吴家林; 沙丹; 马广源; 季亚勇; 孙燕萍; 张敬平

    2011-01-01

    Objective:To establish a multiplex PCR technique for rapid and specific detection of Campylobacter jejuni. Methods: Two sets of primers from map A and hipO gene of Campylobacter jejuni were selected and added into one amplification system to perform PCR. The system was optimized, the specificity and sensitivity of this system were evaluated, and artificially - contaminated chicken were detected. Results: The assay was designed to amplify the 589 bp and 323 bp regions of corresponding genes map A and hipO with high sensitivity and specificity. Sensitivity of the assay was 105 cfu/ml for bacteria samples and 101 cfu/ml for Campylobacter jejuni in artificially - contaminated chicken with incubation at 421 for 36 hours. Conclusion: A rapid, specific and sensitive multiplex PCR technique for the simultaneous detection of Campylobacter jejuni has been studied primarily.%目的:建立能快速、特异检测空肠弯曲菌的多重PCR技术.方法:选用针对空肠弯曲菌外膜蛋白A(mapA)基因和马尿酸酶(hipO)基因的2对引物,在同一扩增体系中进行PCR,优化反应体系,测定特异性和灵敏度,并进行了鸡肉模拟样品检测.结果:该方法扩增目的基因片段分别为589 bp和323 bp,特异性和灵敏度均高.细菌纯培养物的检测灵敏度为105 cfu/ml,鸡肉模拟样品42℃预增菌36 h后检测灵敏度能达到101 cfu/ml.结论:初步建立能快速、灵敏、特异地测定空肠弯曲菌的多重PCR检测技术.

  9. The CsrA-FliW network controls polar localization of the dual-function flagellin mRNA in Campylobacter jejuni

    OpenAIRE

    Dugar, Gaurav; Svensson, Sarah L.; Bischler, Thorsten; Wäldchen, Sina; Reinhardt, Richard; Sauer, Markus; Sharma, Cynthia M.

    2016-01-01

    The widespread CsrA/RsmA protein regulators repress translation by binding GGA motifs in bacterial mRNAs. CsrA activity is primarily controlled through sequestration by multiple small regulatory RNAs. Here we investigate CsrA activity control in the absence of antagonizing small RNAs by examining the CsrA regulon in the human pathogen Campylobacter jejuni. We use genome-wide co-immunoprecipitation combined with RNA sequencing to show that CsrA primarily binds flagellar mRNAs and identify the ...

  10. Direct Quantification of Campylobacter jejuni in Chicken Fecal Samples Using Real-Time PCR: Evaluation of Six Rapid DNA Extraction Methods

    DEFF Research Database (Denmark)

    Garcia Clavero, Ana Belén; Kamara, Judy N.; Vigre, Håkan;

    2013-01-01

    Direct and accurate quantification of Campylobacter in poultry is crucial for the assessment of public health risks and the evaluation of the effectiveness of control measures against Campylobacter in poultry. The aim of this study was to assess several rapid DNA extraction methods...... DNA extraction methods were compared based on their limit of detection, efficiency, reproducibility, and precision. Standard curves were designed for all the methods tested in order to assess their performance on the direct quantification of C. jejuni in chicken fecal samples. As a result...... efficiency was not optimal (AE = 139.5 %). DNA extraction methods Easy-DNA Invitrogen, MiniMAG® and NucleoSpin® Tissue produced good real-time PCR reproducibility generating standard deviations from 0.3 to 0.8 between replicates....

  11. Campylobacter isolation from the feces of sheep with a history of reproductive disorders bred in the state of São Paulo, Brazil

    Directory of Open Access Journals (Sweden)

    Huber Rizzo

    2015-12-01

    Full Text Available Campylobacter species are a significant cause of sheep abortion in most sheep-raising countries. The relationship between the presence of Campylobacter spp. in fecal samples and reproductive disorders was investigated in 274 sheep from 28 properties in the state of São Paulo, Brazil. Biological samples from 16 aborted fetuses, one uterus, six placentas, five uterine secretion samples, five vaginal swabs, 17 semen samples, and three preputial swabs were also subjected to bacterial isolation. The bacteria were isolated from fecal samples of 14.9% (5/28 of the properties, affecting 3.65% (10/274 of the sheep, 3.5% (9/255 of females and 5.3% (1/19 of males. Campylobacter jejuni was the most prevalent species, present in 66.67% (7 of the positive samples, followed by Campylobacter coli, present in 22.22% (2, and one strain was identified as Campylobacter spp. The birth of “weak” lambs (p=0.06, OR=6.83 and CI=1.73 to 27.05 and neonatal death (p=0.087, OR=3.5 and CI=0.83 to 14.72 were associated with the fecal isolation of Campylobacter spp. Diarrhea was also associated with the bacteria (p=0.003, OR=9.83 and CI=2.19 to 44.18. The dissemination of Campylobacter spp. in Brazilian sheep is low and that, at present, the existing strains are not responsible for significant economic losses in sheep production, especially in adult animals.

  12. Multiple factors interact to produce responses resembling spectrum of human disease in Campylobacter jejuni infected C57BL/6 IL-10-/- mice

    Directory of Open Access Journals (Sweden)

    Wolf John E

    2009-03-01

    Full Text Available Abstract Background Campylobacter jejuni infection produces a spectrum of clinical presentations in humans – including asymptomatic carriage, watery diarrhea, and bloody diarrhea – and has been epidemiologically associated with subsequent autoimmune neuropathies. This microorganism is genetically variable and possesses genetic mechanisms that may contribute to variability in nature. However, relationships between genetic variation in the pathogen and variation in disease manifestation in the host are not understood. We took a comparative experimental approach to explore differences among different C. jejuni strains and studied the effect of diet on disease manifestation in an interleukin-10 deficient mouse model. Results In the comparative study, C57BL/6 interleukin-10-/- mice were infected with seven genetically distinct C. jejuni strains. Four strains colonized the mice and caused disease; one colonized with no disease; two did not colonize. A DNA:DNA microarray comparison of the strain that colonized mice without disease to C. jejuni 11168 that caused disease revealed that putative virulence determinants, including loci encoding surface structures known to be involved in C. jejuni pathogenesis, differed from or were absent in the strain that did not cause disease. In the experimental study, the five colonizing strains were passaged four times in mice. For three strains, serial passage produced increased incidence and degree of pathology and decreased time to develop pathology; disease shifted from watery to bloody diarrhea. Mice kept on an ~6% fat diet or switched from an ~12% fat diet to an ~6% fat diet just before infection with a non-adapted strain also exhibited increased incidence and severity of disease and decreased time to develop disease, although the effects of diet were only statistically significant in one experiment. Conclusion C. jejuni strain genetic background and adaptation of the strain to the host by serial passage

  13. Impact of Campylobacter jejuni cj0268c knockout mutation on intestinal colonization, translocation, and induction of immunopathology in gnotobiotic IL-10 deficient mice.

    Directory of Open Access Journals (Sweden)

    Markus M Heimesaat

    Full Text Available BACKGROUND: Although Campylobacter jejuni infections have a high prevalence worldwide and represent a significant socioeconomic burden, the underlying molecular mechanisms of induced intestinal immunopathology are still not well understood. We have recently generated a C. jejuni mutant strain NCTC11168::cj0268c, which has been shown to be involved in cellular adhesion and invasion. The immunopathological impact of this gene, however, has not been investigated in vivo so far. METHODOLOGY/PRINCIPAL FINDINGS: Gnotobiotic IL-10 deficient mice were generated by quintuple antibiotic treatment and perorally infected with C. jejuni mutant strain NCTC11168::cj0268c, its complemented version (NCTC11168::cj0268c-comp-cj0268c, or the parental strain NCTC11168. Kinetic analyses of fecal pathogen loads until day 6 post infection (p.i. revealed that knockout of cj0268c did not compromise intestinal C. jejuni colonization capacities. Whereas animals irrespective of the analysed C. jejuni strain developed similar clinical symptoms of campylobacteriosis (i.e. enteritis, mice infected with the NCTC11168::cj0268c mutant strain displayed significant longer small as well as large intestinal lengths indicative for less distinct C. jejuni induced pathology when compared to infected control groups at day 6 p.i. This was further supported by significantly lower apoptotic and T cell numbers in the colonic mucosa and lamina propria, which were paralleled by lower intestinal IFN-γ and IL-6 concentrations at day 6 following knockout mutant NCTC11168::cj0268c as compared to parental strain infection. Remarkably, less intestinal immunopathology was accompanied by lower IFN-γ secretion in ex vivo biopsies taken from mesenteric lymphnodes of NCTC11168::cj0268c infected mice versus controls. CONCLUSION/SIGNIFICANCE: We here for the first time show that the cj0268c gene is involved in mediating C. jejuni induced immunopathogenesis in vivo. Future studies will provide further

  14. Structural and Functional Analysis of Campylobacter jejuni PseG: a Udp-sugarhydrolase from the Pseudaminic Acid Biosynthetic Pathway

    Energy Technology Data Exchange (ETDEWEB)

    E Rangarajan; A Proteau; Q Cui; S Logan; Z Potetinova; D Whitfield; E Purisima; M Cygler; A Matte; et al.

    2011-12-31

    Flagella of the bacteria Helicobacter pylori and Campylobacter jejuni are important virulence determinants, whose proper assembly and function are dependent upon glycosylation at multiple positions by sialic acid-like sugars, such as 5,7-diacetamido-3,5,7,9-tetradeoxy-l-glycero-l-manno-nonulosonic acid (pseudaminic acid (Pse)). The fourth enzymatic step in the pseudaminic acid pathway, the hydrolysis of UDP-2,4-diacetamido-2,4,6-trideoxy-{beta}-l-altropyranose to generate 2,4-diacetamido-2,4,6-trideoxy-l-altropyranose, is performed by the nucleotide sugar hydrolase PseG. To better understand the molecular basis of the PseG catalytic reaction, we have determined the crystal structures of C. jejuni PseG in apo-form and as a complex with its UDP product at 1.8 and 1.85 {angstrom} resolution, respectively. In addition, molecular modeling was utilized to provide insight into the structure of the PseG-substrate complex. This modeling identifies a His{sup 17}-coordinated water molecule as the putative nucleophile and suggests the UDP-sugar substrate adopts a twist-boat conformation upon binding to PseG, enhancing the exposure of the anomeric bond cleaved and favoring inversion at C-1. Furthermore, based on these structures a series of amino acid substitution derivatives were constructed, altering residues within the active site, and each was kinetically characterized to examine its contribution to PseG catalysis. In conjunction with structural comparisons, the almost complete inactivation of the PseG H17F and H17L derivatives suggests that His{sup 17} functions as an active site base, thereby activating the nucleophilic water molecule for attack of the anomeric C-O bond of the UDP-sugar. As the PseG structure reveals similarity to those of glycosyltransferase family-28 members, in particular that of Escherichia coli MurG, these findings may also be of relevance for the mechanistic understanding of this important enzyme family.

  15. Antimicrobial susceptibility of Campylobacter sp strains isolated from calves with and without diarrhea in Minas Gerais state, Brazil Susceptibilidade a antimicrobianos de amostras de Campylobacter sp isoladas de bezerros com e sem diarréia, no estado de Minas Gerais, Brasil

    Directory of Open Access Journals (Sweden)

    Karina Leite Miranda

    2007-06-01

    Full Text Available The antimicrobial susceptibility of 25 Campylobacter sp strains isolated from calves with and without diarrhea - 7 C. coli, 16 C. fetus and 2 C. jejuni was studied by the disk diffusion method. Eleven antimicrobial agents were tested amikacin, ampicillin, kanamycin, chloramphenicol, erythromycin, gentamicin, neomycin, nitrofurantoin, penicillin G, tetracycline and sulfamethoxazole-trimethoprim. All Campylobacter sp strains were susceptible to amikacin, ampicillin, chloramphenicol, erythromycin, gentamicin, neomycin and nitrofurantoin. Three strains were moderately susceptible to kanamycin (2 C. coli and 1 C. fetus. All the strains were resistant to penicillin G. Two C. fetus strains were moderately susceptible to sulfamethoxazole-trimethoprim and 1 C. coli, 9 C. fetus and 2 C. jejuni strains were resistant. Two C. fetus strains were moderately susceptible to tetracycline and 3 C. coli, 2 C. fetus and 1 C. jejuni strains were resistant. Eleven strains showed multidrug resistance (2 C. coli, 8 C. fetus and 1 C. jejuni. There was no correlation between resistance of Campylobacter sp strains to antimicrobials and the occurrence of diarrhea in calves. The frequency of resistance and, most importantly, multi drug resistance found among Campylobacter sp strains isolated from calves in Minas Gerais, Brazil, were high and the patterns of resistance observed are related to the antimicrobials agents most largely used in cattle in Brazil.Foi estudado o perfil de susceptibilidade aos antimicrobianos de 25 amostras de Campylobacter sp isoladas de bezerros com e sem diarréia (7 C. coli, 16 C. fetus e 2 C. jejuni. Foram testados pelo método de difusão 11 agentes antimicrobianos: amicacina, ampicilina, canamicina, cloranfenicol, eritromicina, gentamicina, neomicina, nitrofurantoína, penicilina G, tetraciclina e sulfametoxazole-trimetoprim. Todas as amostras de Campylobacter sp foram susceptíveis a amicacina, ampicilina, cloranfenicol, eritromicina

  16. Monomerization alters the dynamics of the lid region in Campylobacter jejuni CstII: an MD simulation study.

    Science.gov (United States)

    Prabhakar, Pradeep Kumar; Srivastava, Alka; Rao, K Krishnamurthy; Balaji, Petety V

    2016-04-01

    CstII, a bifunctional (α2,3/8) sialyltransferase from Campylobacter jejuni, is a homotetramer. It has been reported that mutation of the interface residues Phe121 (F121D) or Tyr125 (Y125Q) leads to monomerization and partial loss of enzyme activity, without any change in the secondary or tertiary structures. MD simulations of both tetramer and monomer, with and without bound donor substrate, were performed for the two mutants and WT to understand the reasons for partial loss of activity due to monomerization since the active site is located within each monomer. RMSF values were found to correlate with the crystallographic B-factor values indicating that the simulations are able to capture the flexibility of the molecule effectively. There were no gross changes in either the secondary or tertiary structure of the proteins during MD simulations. However, interface is destabilized by the mutations, and more importantly the flexibility of the lid region (Gly152-Lys190) is affected. The lid region accesses three major conformations named as open, intermediate, and closed conformations. In both Y121Q and F121D mutants, the closed conformation is accessed predominantly. In this conformation, the catalytic base His188 is also displaced. Normal mode analysis also revealed differences in the lid movement in tetramer and monomer. This provides a possible explanation for the partial loss of enzyme activity in both interface mutants. The lid region controls the traffic of substrates and products in and out of the active site, and the dynamics of this region is regulated by tetramerization. Thus, this study provides valuable insights into the role of loop dynamics in enzyme activity of CstII. PMID:26208676

  17. Antiparasitic drug nitazoxanide inhibits the pyruvate oxidoreductases of Helicobacter pylori, selected anaerobic bacteria and parasites, and Campylobacter jejuni.

    Science.gov (United States)

    Hoffman, Paul S; Sisson, Gary; Croxen, Matthew A; Welch, Kevin; Harman, W Dean; Cremades, Nunilo; Morash, Michael G

    2007-03-01

    Nitazoxanide (NTZ) exhibits broad-spectrum activity against anaerobic bacteria and parasites and the ulcer-causing pathogen Helicobacter pylori. Here we show that NTZ is a noncompetitive inhibitor (K(i), 2 to 10 microM) of the pyruvate:ferredoxin/flavodoxin oxidoreductases (PFORs) of Trichomonas vaginalis, Entamoeba histolytica, Giardia intestinalis, Clostridium difficile, Clostridium perfringens, H. pylori, and Campylobacter jejuni and is weakly active against the pyruvate dehydrogenase of Escherichia coli. To further mechanistic studies, the PFOR operon of H. pylori was cloned and overexpressed in E. coli, and the multisubunit complex was purified by ion-exchange chromatography. Pyruvate-dependent PFOR activity with NTZ, as measured by a decrease in absorbance at 418 nm (spectral shift from 418 to 351 nm), unlike the reduction of viologen dyes, did not result in the accumulation of products (acetyl coenzyme A and CO(2)) and pyruvate was not consumed in the reaction. NTZ did not displace the thiamine pyrophosphate (TPP) cofactor of PFOR, and the 351-nm absorbing form of NTZ was inactive. Optical scans and (1)H nuclear magnetic resonance analyses determined that the spectral shift (A(418) to A(351)) of NTZ was due to protonation of the anion (NTZ(-)) of the 2-amino group of the thiazole ring which could be generated with the pure compound under acidic solutions (pK(a) = 6.18). We propose that NTZ(-) intercepts PFOR at an early step in the formation of the lactyl-TPP transition intermediate, resulting in the reversal of pyruvate binding prior to decarboxylation and in coordination with proton transfer to NTZ. Thus, NTZ might be the first example of an antimicrobial that targets the "activated cofactor" of an enzymatic reaction rather than its substrate or catalytic sites, a novel mechanism that may escape mutation-based drug resistance. PMID:17158936

  18. Identification of unusual Campylobacter-like isolates from poultry products as Helicobacter pullorum

    DEFF Research Database (Denmark)

    Atabay, H.I.; Corry, J.E.L.; On, Stephen L.W.

    1998-01-01

    Campy identification kits and a 16-test identification scheme developed for campylobacters failed to identify these bacteria, or identified them as Campylobacter spp. Eighteen strains (including the two isolated on a different occasion) were chosen for examination using a more comprehensive......Twenty-six unclassified Campylobacter-like strains previously isolated from 15 chicken carcasses and caecal contents, together with two more strains isolated from chicken faeces on a different occasion, were identified as Helicobacter pullorum using various phenotypic identification methods. API...

  19. Variation in the limit-of-detection of the ProSpecT Campylobacter microplate enzyme immunoassay in stools spiked with emerging Campylobacter species.

    Science.gov (United States)

    Bojanić, Krunoslav; Midwinter, Anne Camilla; Marshall, Jonathan Craig; Rogers, Lynn Elizabeth; Biggs, Patrick Jon; Acke, Els

    2016-08-01

    Campylobacter enteritis in humans is primarily associated with C. jejuni/coli infection. The impact of other Campylobacter spp. is likely to be underestimated due to the bias of culture methods towards Campylobacter jejuni/coli diagnosis. Stool antigen tests are becoming increasingly popular and appear generally less species-specific. A review of independent studies of the ProSpecT® Campylobacter Microplate enzyme immunoassay (EIA) developed for C. jejuni/coli showed comparable diagnostic results to culture methods but the examination of non-jejuni/coli Campylobacter spp. was limited and the limit-of-detection (LOD), where reported, varied between studies. This study investigated LOD of EIA for Campylobacter upsaliensis, Campylobacter hyointestinalis and Campylobacter helveticus spiked in human stools. Multiple stools and Campylobacter isolates were used in three different concentrations (10(4)-10(9)CFU/ml) to reflect sample heterogeneity. All Campylobacter species evaluated were detectable by EIA. Multivariate analysis showed LOD varied between Campylobacter spp. and faecal consistency as fixed effects and individual faecal samples as random effects. EIA showed excellent performance in replicate testing for both within and between batches of reagents, in agreement between visual and spectrophotometric reading of results, and returned no discordance between the bacterial concentrations within independent dilution test runs (positive results with lower but not higher concentrations). This study shows how limitations in experimental procedures lead to an overestimation of consistency and uniformity of LOD for EIA that may not hold under routine use in diagnostic laboratories. Benefits and limitations for clinical practice and the influence on estimates of performance characteristics from detection of multiple Campylobacter spp. by EIA are discussed.

  20. Variation in the limit-of-detection of the ProSpecT Campylobacter microplate enzyme immunoassay in stools spiked with emerging Campylobacter species.

    Science.gov (United States)

    Bojanić, Krunoslav; Midwinter, Anne Camilla; Marshall, Jonathan Craig; Rogers, Lynn Elizabeth; Biggs, Patrick Jon; Acke, Els

    2016-08-01

    Campylobacter enteritis in humans is primarily associated with C. jejuni/coli infection. The impact of other Campylobacter spp. is likely to be underestimated due to the bias of culture methods towards Campylobacter jejuni/coli diagnosis. Stool antigen tests are becoming increasingly popular and appear generally less species-specific. A review of independent studies of the ProSpecT® Campylobacter Microplate enzyme immunoassay (EIA) developed for C. jejuni/coli showed comparable diagnostic results to culture methods but the examination of non-jejuni/coli Campylobacter spp. was limited and the limit-of-detection (LOD), where reported, varied between studies. This study investigated LOD of EIA for Campylobacter upsaliensis, Campylobacter hyointestinalis and Campylobacter helveticus spiked in human stools. Multiple stools and Campylobacter isolates were used in three different concentrations (10(4)-10(9)CFU/ml) to reflect sample heterogeneity. All Campylobacter species evaluated were detectable by EIA. Multivariate analysis showed LOD varied between Campylobacter spp. and faecal consistency as fixed effects and individual faecal samples as random effects. EIA showed excellent performance in replicate testing for both within and between batches of reagents, in agreement between visual and spectrophotometric reading of results, and returned no discordance between the bacterial concentrations within independent dilution test runs (positive results with lower but not higher concentrations). This study shows how limitations in experimental procedures lead to an overestimation of consistency and uniformity of LOD for EIA that may not hold under routine use in diagnostic laboratories. Benefits and limitations for clinical practice and the influence on estimates of performance characteristics from detection of multiple Campylobacter spp. by EIA are discussed. PMID:27317896

  1. Expression, purification, crystallization and preliminary X-ray characterization of two crystal forms of stationary-phase survival E protein from Campylobacter jejuni

    Energy Technology Data Exchange (ETDEWEB)

    Gonçalves, A. M. D.; Rêgo, A. T. [Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa, Avenida da República, Apartado 127, 2781-901 Oeiras (Portugal); Thomaz, M. [Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa, Avenida da República, Apartado 127, 2781-901 Oeiras (Portugal); Instituto de Biologia Experimental e Tecnológica, Apartado 12, 2748-901 Oeiras (Portugal); Enguita, F. J., E-mail: fenguita@fm.ul.pt [Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa, Avenida da República, Apartado 127, 2781-901 Oeiras (Portugal); Instituto de Medicina Molecular, Faculdade de Medicina, Universidade de Lisboa, Avenida Professor Egas Moniz, 1649-028 Lisboa (Portugal); Carrondo, M. A. [Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa, Avenida da República, Apartado 127, 2781-901 Oeiras (Portugal)

    2008-03-01

    Survival E (SurE) protein from Campylobacter jejuni, a Gram-negative mesophile, has been overexpressed in Escherichia coli as a soluble protein, successfully purified and crystallized in two distinct crystal forms. Survival E (SurE) protein from Campylobacter jejuni, a Gram-negative mesophile, has been overexpressed in Escherichia coli as a soluble protein, successfully purified and crystallized in two distinct crystal forms. The first form belongs to space group P2{sub 1}2{sub 1}2{sub 1}, with a tetramer in the asymmetric unit and unit-cell parameters a = 80.5, b = 119.0, c = 135.3 Å. The second form belongs to space group C2, with unit-cell parameters a = 121.4, b = 47.1, c = 97.8 Å, and contains a dimer in the asymmetric unit. Diffraction data have been collected from these crystal forms to 2.5 and 2.95 Å resolution, respectively.

  2. Establishment and application of loop-mediated isothermal amplification method for rapid detection of Campylobacter jejuni%空肠弯曲杆菌LAMP检测方法的建立及初步应用

    Institute of Scientific and Technical Information of China (English)

    唐梦君; 周生; 张小燕; 唐修君; 顾荣; 高玉时

    2012-01-01

    In order to establish a method of loop-mediated isothermal amplification (LAMP) for detection of Campylobacter jejuni , specific loop-mediated isothermal amplification primers were designed, and a novel and highly specific loop-mediated isothermal amplification assay for the sensitive and rapid detection of Campylobacter jejuni were developed. According to Campylobacter jejuni gyrA gene sequences published on GenBank, the natural infection rate of Campylobacter jejuni was tested in ten chicken breeds. The results show that the products of LAMP demonstrated the typical ladder patterns and digested fragments were found with the predicted sizes. Sensitivity of the LAMP assay for direct detection of Campylobacter jejuni in pure cultures was 20 cfu/mL. It was 10-fold more sensitive than that of the conventional PCR assay. The assay identified Campylobacter jejuni correctly, but did not detect 7 non-Campylobacter jejuni strains. The natural infection rate of Campylobacter jejuni was 6%-90% in the ten chicken breeds. The LAMP assay is a sensitive, rapid and simple tool for the detection of Campylobacter jejuni and will play a role in clinical detection.%目的 建立一种灵敏的环介导等温扩增方法(Loop-mediated Isothermal Amplification,LAMP)用于空肠弯曲杆菌(Campylobacter jejuni)的快速检测.方法 根据已公布的空肠弯曲杆菌旋转酶基因(gyrA)设计引物,建立并优化LAMP反应体系,对检测方法的特异性和灵敏度进行验证,并运用该方法对我国不同地方鸡种空肠弯曲杆菌的携带率进行调查.结果 该方法能扩增出典型的梯形条带,且扩增产物的酶切鉴定结果与理论值相符;对单增李斯特菌等8株实验菌株进行检测,仅空肠弯曲杆菌的LAMP结果为阳性;该方法检测空肠弯曲杆菌纯培养物的灵敏度为20 cfu/mL,高于常规PCR;不同地方鸡种空肠弯曲杆菌携带率介于6%~90%之间,差异显著.结论 本试验建立的空肠弯曲杆菌LAMP检测

  3. Sequence variability of Campylobacter temperate bacteriophages

    Directory of Open Access Journals (Sweden)

    Ng Lai-King

    2008-03-01

    Full Text Available Abstract Background Prophages integrated within the chromosomes of Campylobacter jejuni isolates have been demonstrated very recently. Prior work with Campylobacter temperate bacteriophages, as well as evidence from prophages in other enteric bacteria, suggests these prophages might have a role in the biology and virulence of the organism. However, very little is known about the genetic variability of Campylobacter prophages which, if present, could lead to differential phenotypes in isolates carrying the phages versus those that do not. As a first step in the characterization of C. jejuni prophages, we investigated the distribution of prophage DNA within a C. jejuni population assessed the DNA and protein sequence variability within a subset of the putative prophages found. Results Southern blotting of C. jejuni DNA using probes from genes within the three putative prophages of the C. jejuni sequenced strain RM 1221 demonstrated the presence of at least one prophage gene in a large proportion (27/35 of isolates tested. Of these, 15 were positive for 5 or more of the 7 Campylobacter Mu-like phage 1 (CMLP 1, also designated Campylobacter jejuni integrated element 1, or CJIE 1 genes tested. Twelve of these putative prophages were chosen for further analysis. DNA sequencing of a 9,000 to 11,000 nucleotide region of each prophage demonstrated a close homology with CMLP 1 in both gene order and nucleotide sequence. Structural and sequence variability, including short insertions, deletions, and allele replacements, were found within the prophage genomes, some of which would alter the protein products of the ORFs involved. No insertions of novel genes were detected within the sequenced regions. The 12 prophages and RM 1221 had a % G+C very similar to C. jejuni sequenced strains, as well as promoter regions characteristic of C. jejuni. None of the putative prophages were successfully induced and propagated, so it is not known if they were functional or

  4. Campylobacter in broiler slaughter samples assessed by direct count on mCCDA and Campy-Cefex agar.

    Science.gov (United States)

    Gonsalves, Camila Cristina; Borsoi, Anderlise; Perdoncini, Gustavo; Rodrigues, Laura Beatriz; do Nascimento, Vladimir Pinheiro

    2016-01-01

    Campylobacter spp. cause foodborne illnesses in humans primarily through the consumption of contaminated chicken. The aim of this study was to evaluate the United States Department of Agriculture's (USDA) recommended methodology, protocol MLG 41.02, for the isolation, identification and direct plate counting of Campylobacter jejuni and C. coli samples from the broiler slaughtering process. A plating method using both mCCDA and Campy-Cefex agars is recommended to recover Campylobacter cells. It is also possible to use this method in different matrices (cloacal swabs and water samples). Cloacal swabs, samples from pre-chiller and post-chiller carcasses and samples of pre-chiller, chiller and direct supply water were collected each week for four weeks from the same flock at a slaughterhouse located in an abattoir in southern Brazil. Samples were analyzed to directly count Campylobacter spp., and the results showed a high frequency of Campylobacter spp. on Campy-Cefex agar. For the isolated species, 72% were identified as Campylobacter jejuni and 38% as Campylobacter coli. It was possible to count Campylobacter jejuni and Campylobacter coli from different samples, including the water supply samples, using the two-agar method. These results suggest that slaughterhouses can use direct counting methods with both agars and different matrices as a monitoring tool to assess the presence of Campylobacter bacteria in their products. PMID:27237112

  5. Exploiting the Campylobacter jejuni protein glycosylation system for glycoengineering vaccines and diagnostic tools directed against brucellosis

    Directory of Open Access Journals (Sweden)

    Iwashkiw Jeremy A

    2012-01-01

    Full Text Available Abstract Background Immune responses directed towards surface polysaccharides conjugated to proteins are effective in preventing colonization and infection of bacterial pathogens. Presently, the production of these conjugate vaccines requires intricate synthetic chemistry for obtaining, activating, and attaching the polysaccharides to protein carriers. Glycoproteins generated by engineering bacterial glycosylation machineries have been proposed to be a viable alternative to traditional conjugation methods. Results In this work we expressed the C. jejuni oligosaccharyltansferase (OTase PglB, responsible for N-linked protein glycosylation together with a suitable acceptor protein (AcrA in Yersinia enterocolitica O9 cells. MS analysis of the acceptor protein demonstrated the transfer of a polymer of N-formylperosamine to AcrA in vivo. Because Y. enterocolitica O9 and Brucella abortus share an identical O polysaccharide structure, we explored the application of the resulting glycoprotein in vaccinology and diagnostics of brucellosis, one of the most common zoonotic diseases with over half a million new cases annually. Injection of the glycoprotein into mice generated an IgG response that recognized the O antigen of Brucella, although this response was not protective against a challenge with a virulent B. abortus strain. The recombinant glycoprotein coated onto magnetic beads was efficient in differentiating between naïve and infected bovine sera. Conclusion Bacterial engineered glycoproteins show promising applications for the development on an array of diagnostics and immunoprotective opportunities in the future.

  6. Association of Anti-GT1a Antibodies with an Outbreak of Guillain-Barre Syndrome and Analysis of Ganglioside Mimicry in an Associated Campylobacter jejuni Strain.

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    Maojun Zhang

    Full Text Available An outbreak of Guillain-Barré syndrome (GBS, subsequent to Campylobacter jejuni enteritis, occurred in China in 2007. Serum anti-ganglioside antibodies were measured in GBS patients and controls. Genome sequencing was used to determine the phylogenetic relationship among three C. jejuni strains from a patient with GBS (ICDCCJ07001, a patient with gastroenteritis (ICDCCJ07002 and a healthy carrier (ICDCCJ07004, which were all associated with the outbreak. The ganglioside-like structures of the lipo-oligosaccharides of these strains were determined by mass spectrometry. Seventeen (53% of the GBS patients had anti-GT1a IgG antibodies. GT1a mimicry was found in the lipo-oligosaccharides of strain ICDCCJ07002 and ICDCCJ07004; but a combination of GM3/GD3 mimics was observed in ICDCCJ07001, although this patient had anti-GT1a IgG antibodies. A single-base deletion in a glycosyltransferase gene caused the absence of GT1a mimicry in ICDCCJ07001. The phylogenetic tree showed that ICDCCJ07002 and ICDCCJ07004 were genetically closer to each other than to ICDCCJ07001. C. jejuni, bearing a GT1a-like lipo-oligosaccharide, might have caused the GBS outbreak and the loss of GT1a mimicry may have helped ICDCCJ07001 to survive in the host.

  7. Adaptation of Campylobacter jejuni to biocides used in the food industry affects biofilm structure, adhesion strength, and cross-resistance to clinical antimicrobial compounds.

    Science.gov (United States)

    Techaruvichit, Punnida; Takahashi, Hajime; Kuda, Takashi; Miya, Satoko; Keeratipibul, Suwimon; Kimura, Bon

    2016-08-01

    The emergence of biocide-adapted Campylobacter jejuni strains that developed into biofilms and their potential to develop clinical resistance to antimicrobial compounds was studied. C. jejuni was grown in sub-lethal concentrations of five biocides used in the food industry. C. jejuni exhibited adaptation to these biocides with increased minimum inhibitory concentrations. The 3-D structures of the biofilms produced by the biocide-adapted cells were investigated by atomic force microscopy (AFM). The results revealed marked variability in biofilm architecture, including ice-crystal-like structures. Adaptation to the biocides enhanced biofilm formation, with significant increases in biovolume, surface coverage, roughness, and the surface adhesion force of the biofilms. Adaptation to commercial biocides induced resistance to kanamycin and streptomycin. This study suggests that the inappropriate use of biocides may lead to cells being exposed to them at sub-lethal concentrations, which can result in adaptation of the pathogens to the biocides and a subsequent risk to public health. PMID:27353218

  8. Campylobacter spp. Zoonotic micro organism

    OpenAIRE

    Ivanović Snežana

    2008-01-01

    There are two species of Campylobacter but, for human and animal health the most important are Campylobacter jejuni, Campylobacter coli, and for animal species also Campylobacter fetus and Campylobacter laridis. Different temperatures, drying, pH of the environment, disinfectants, spices and probiotics and antibiotics influence the growth and multiplication of this micro organism in the animal as well as in the animal products. Campylobacter is present everywhere in the nature: in water, soil...

  9. Campylobacter spp. and birds of prey.

    Science.gov (United States)

    Dipineto, Ludovico; De Luca Bossa, Luigi Maria; Russo, Tamara Pasqualina; Cutino, Eridania Annalisa; Gargiulo, Antonio; Ciccarelli, Francesca; Raia, Pasquale; Menna, Lucia Francesca; Fioretti, Alessandro

    2014-06-01

    A total of 170 birds of prey admitted to two Wildlife Rescue and Rehabilitation Centers of Italy were examined. Birds were divided by diurnal (n = 15) and nocturnal (n = 7) species, sampled by cloacal swabs, and examined for Campylobacter spp. by cultural and molecular methods. Campylobacter spp. were isolated in 43 out of the 170 (25.3%) birds of prey examined. Among these, 43/43 (100%) were identified as Campylobacter jejuni and 10/43 (23.3%) were identified as Campylobacter coli recovered from mixed infections. Diurnal birds of prey showed a significantly higher prevalence value (P = 0.0006) for Campylobacter spp. than did nocturnal birds of prey. PMID:25055637

  10. Chitosan supplementation reduces enteric colonization of Campylobacter jejuni in broiler chickens and down-regulates expression of colonization genes

    Science.gov (United States)

    Campylobacter is one of the leading causes of foodborne bacterial gastroenteritis worldwide, and poultry is considered as the most common source of human infections. Campylobacter is prevalent in most poultry flocks and a reduction of Campylobacter in poultry would greatly reduce the risk of campylo...

  11. THE OCCURRENCE OF PLASMID DNA IN CAMPYLOBACTER JEJUNI AND CAMPYLOBACTER COLI AND ANALYSIS OF RESTRICTION ENDONUCLEASE%空肠弯曲菌质粒的提取鉴定和限制性内切酶水解片段的分析

    Institute of Scientific and Technical Information of China (English)

    孙自捷; 宋后燕; 段恕诚

    1988-01-01

    Thirty-two strains of different serotypes and biotypes of Campylobaoter jejuni and Campylobacter coli isolated from the stool of children and domestic animals were examined for the presence of plasmid DNA Agarose gel electrophoresis of alkaline-extracted DNA showed the occurrence of plasmid bands in 12 strains.Ten of them showed only one plasmid band in>28Kb area of gel,and 7 strains contained this plasmid were further analyzed by restriction endonuclease Hind Ⅲ.All restriction fragments showed heterogeneity.Other 2 of the 12 strains from domestic animals were observed and 3 and 4 plasmid bands were demonstrated in agarose gel respectively.The incidenoe of plasmid in Campylobacter jejuni strains was similar to Campylobacter coli,while the incidence of plasmid in Campylobacter strains from the stool of domestic animals was higher than in children.%使用修改的Brruboim快速碱抽提法,对32株来源不同,和不同血清型和生物型空肠弯曲菌株进行了质粒DNA检测和分析.12株细菌的质粒抽提液在琼脂糖电泳上显示有质粒带,其中10株均仅含有一条>23Kb的质粒,同时7株用限制性内切酶Hind Ⅲ消化后水解片段分析发现这一条>23Kb的质粒为非同源性.其他2株细菌在琼脂糖电泳上显示3和4条质粒带.不同生物型空肠弯曲菌的质粒携带率基本相同,而从家禽分离的空肠弯曲菌株比从儿童分离的菌株质粒携带率要高.

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

    Science.gov (United States)

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

    2016-01-01

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

  13. Description of Campylobacter fetus subsp. testudinum subsp. nov., isolated from humans and reptiles

    Science.gov (United States)

    A polyphasic study was undertaken to determine the taxonomic position of 13 Campylobacter fetus-like isolates from humans (n=8) and reptiles (n=5). Phenotypic characterization, Genusgenus-specific and sap insertion-PCR initially identified all human isolates as type A Campylobacter fetus. Phylogenet...

  14. Phylogenetic diversity and position of the genus Campylobacter

    Science.gov (United States)

    Lau, P. P.; DeBrunner-Vossbrinck, B.; Dunn, B.; Miotto, K.; MacDonnell, M. T.; Rollins, D. M.; Pillidge, C. J.; Hespell, R. B.; Colwell, R. R.; Sogin, M. L.; Fox, G. E.

    1987-01-01

    RNA sequence analysis has been used to examine the phylogenetic position and structure of the genus Campylobacter. A complete 5S rRNA sequence was determined for two strains of Campylobacter jejuni and extensive partial sequences of the 16S rRNA were obtained for several strains of C. jejuni and Wolinella succinogenes. In addition limited partial sequence data were obtained from the 16S rRNAs of isolates of C. coli, C. laridis, C. fetus, C. fecalis, and C. pyloridis. It was found that W. succinogenes is specifically related to, but not included, in the genus Campylobacter as presently constituted. Within the genus significant diversity was noted. C. jejuni, C. coli and C. laridis are very closely related but the other species are distinctly different from one another. C. pyloridis is without question the most divergent of the Campylobacter isolates examined here and is sufficiently distinct to warrant inclusion in a separate genus. In terms of overall position in bacterial phylogeny, the Campylobacter/Wolinella cluster represents a deep branching most probably located within an expanded version of the Division containing the purple photosynthetic bacteria and their relatives. The Campylobacter/Wolinella cluster is not specifically includable in either the alpha, beta or gamma subdivisions of the purple bacteria.

  15. Distribution and Genetic Profiles of Campylobacter in Commercial Broiler Production from Breeder to Slaughter in Thailand.

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    Sakaoporn Prachantasena

    Full Text Available Poultry and poultry products are commonly considered as the major vehicle of Campylobacter infection in humans worldwide. To reduce the number of human cases, the epidemiology of Campylobacter in poultry must be better understood. Therefore, the objective of the present study was to determine the distribution and genetic relatedness of Campylobacter in the Thai chicken production industry. During June to October 2012, entire broiler production processes (i.e., breeder flock, hatchery, broiler farm and slaughterhouse of five broiler production chains were investigated chronologically. Representative isolates of C. jejuni from each production stage were characterized by flaA SVR sequencing and multilocus sequence typing (MLST. Amongst 311 selected isolates, 29 flaA SVR alleles and 17 sequence types (STs were identified. The common clonal complexes (CCs found in this study were CC-45, CC-353, CC-354 and CC-574. C. jejuni isolated from breeders were distantly related to those isolated from broilers and chicken carcasses, while C. jejuni isolates from the slaughterhouse environment and meat products were similar to those isolated from broiler flocks. Genotypic identification of C. jejuni in slaughterhouses indicated that broilers were the main source of Campylobacter contamination of chicken meat during processing. To effectively reduce Campylobacter in poultry meat products, control and prevention strategies should be aimed at both farm and slaughterhouse levels.

  16. PCR技术检测空肠弯曲菌的研究进展%Advances in detection of Campylobacter jejuni by polymerase chain reaction technology

    Institute of Scientific and Technical Information of China (English)

    陈金; 毕水莲

    2015-01-01

    空肠弯曲菌(Campylobacter jejuni)是一种人畜共患病病原菌,可以使人和动物引发多种疾病。目前,检测C. jejuni采用的国标方法是传统的培养法,但C. jejuni培养条件苛刻,且培养法存在操作繁琐、特异性不强、费时等缺点。聚合酶链式反应(polymerase chain reaction, PCR)以其快速、准确、灵敏度高、特异性强的特点,现已广泛应用于C. jejuni的检测,并成为目前快速检测临床与食品中C. jejuni最常用的的方法。本文综述了近年来利用RCR技术,包括常规PCR、多重PCR、巢式PCR、实时荧光定量PCR、PCR-酶联免疫吸附法、最大几率数-PCR、PCR-限制性片段长度多态性、PCR-变性高效液相色谱、PCR-变性梯度凝胶电泳和磁捕获-荧光PCR方法检测C. jejuni的研究进展,并针对这些PCR技术的原理、检测效果、优点和缺点等方面进行了分析比较,为有效控制和预防该菌引起的疾病提供重要信息。%ABSTRACT:As a zoonotic pathogen, Campylobacter jejuni could cause lots of diseases of human and animal. At present, plate-culture method is used to detect C. jejuni as the national standard. But the cultivation conditions of C. jejuni are very strict, and the plate-culture method has the disadvantages such as complicated operation, specificity, and time consuming. Polymerase chain reaction, which is excellent for the rapid, accurate, high sensitivity and specificity, has been widely used in the detection of C. jejuni, and become the most common method for the rapid detection of C. jejuni in clinical and food samples. The recent research progress was summarized in this paper and the principle, effect and characteristics among conventional PCR, multiplex PCR, nested PCR, real-time PCR, PCR-ELISA, MPN (most probable number)-PCR, PCR-RFLP (restriction fragment length polymorphism), PCR-DHPLC (denaturing high performance liquid chromatography), PCR-DGGE (denaturing gradient gel electrophoresis) and IMC

  17. Description and sources of contamination by Campylobacter spp. of river water destined for human consumption in Brittany, France.

    Science.gov (United States)

    Denis, M; M Tanguy; Chidaine, B; Laisney, M-J; Mégraud, F; Fravalo, P

    2011-10-01

    Presence or absence of Campylobacter spp. in water of five rivers upstream from an intake point for drinking water production was investigated, and isolates genetically compared with human, pig and poultry isolates in order to determine their source. River water and drinking water obtained from these rivers were sampled one time per month, over a period of one year, and tested for Campylobacter. Isolates were typed by PFGE. Campylobacter was not detected in treated drinking water, but 50% of the river samples were contaminated. Contamination was observed on the four seasons. In total, 297 Campylobacter isolates were collected and generated 46 PFGE profiles. Campylobacter jejuni was the most frequently detected species in samples (74.1% of the isolates), followed by Campylobacter coli (17.8%) and Campylobacter lari (8.1%). Forty-two of the 46 PFGE profiles were unique. Only one genotype was detected three times in a river during the year and four genotypes in two different rivers. When compared to animal and human Campylobacter PFGE profiles, 14, 11 and one Campylobacter genotypes from water were genetically closed to human, poultry, and pig Campylobacter genotypes, respectively. The Campylobacter population displayed a high level of genetic diversity, suggesting that contamination originated from various origins. Human, poultry and pig were sources of contamination of the river by Campylobacter. Finally, no Campylobacter were detected in drinking water, indicating that the risk of outbreaks due to consumption of drinking water is low.

  18. Multicenter Evaluation of Clinical Diagnostic Methods for Detection and Isolation of Campylobacter spp. from Stool.

    Science.gov (United States)

    Fitzgerald, Collette; Patrick, Mary; Gonzalez, Anthony; Akin, Joshua; Polage, Christopher R; Wymore, Kate; Gillim-Ross, Laura; Xavier, Karen; Sadlowski, Jennifer; Monahan, Jan; Hurd, Sharon; Dahlberg, Suzanne; Jerris, Robert; Watson, Renee; Santovenia, Monica; Mitchell, David; Harrison, Cassandra; Tobin-D'Angelo, Melissa; DeMartino, Mary; Pentella, Michael; Razeq, Jafar; Leonard, Celere; Jung, Carrianne; Achong-Bowe, Ria; Evans, Yaaqobah; Jain, Damini; Juni, Billie; Leano, Fe; Robinson, Trisha; Smith, Kirk; Gittelman, Rachel M; Garrigan, Charles; Nachamkin, Irving

    2016-05-01

    The use of culture-independent diagnostic tests (CIDTs), such as stool antigen tests, as standalone tests for the detection of Campylobacter in stool is increasing. We conducted a prospective, multicenter study to evaluate the performance of stool antigen CIDTs compared to culture and PCR for Campylobacter detection. Between July and October 2010, we tested 2,767 stool specimens from patients with gastrointestinal illness with the following methods: four types of Campylobacter selective media, four commercial stool antigen assays, and a commercial PCR assay. Illnesses from which specimens were positive by one or more culture media or at least one CIDT and PCR were designated "cases." A total of 95 specimens (3.4%) met the case definition. The stool antigen CIDTs ranged from 79.6% to 87.6% in sensitivity, 95.9 to 99.5% in specificity, and 41.3 to 84.3% in positive predictive value. Culture alone detected 80/89 (89.9% sensitivity) Campylobacter jejuni/Campylobacter coli-positive cases. Of the 209 noncases that were positive by at least one CIDT, only one (0.48%) was positive by all four stool antigen tests, and 73% were positive by just one stool antigen test. The questionable relevance of unconfirmed positive stool antigen CIDT results was supported by the finding that noncases were less likely than cases to have gastrointestinal symptoms. Thus, while the tests were convenient to use, the sensitivity, specificity, and positive predictive value of Campylobacter stool antigen tests were highly variable. Given the relatively low incidence of Campylobacter disease and the generally poor diagnostic test characteristics, this study calls into question the use of commercially available stool antigen CIDTs as standalone tests for direct detection of Campylobacter in stool. PMID:26962088

  19. Genotyping of Campylobacter jejuni strains from Danish broiler chickens by restriction fragment length polymorphism of the LPS gene cluster

    DEFF Research Database (Denmark)

    Knudsen, K.N.; Bang, Dang Duong; Nielsen, E.M.;

    2005-01-01

    , the LG genotyping method was used to study the genetic stability of four C. jejuni strains after gastrointestinal passage through experimentally infected chickens. Methods and Results: In the present study, the LG genotyping method was modified with respect to the restriction enzymes used. To validate...... no changes in the LG genotype of the C. jejuni strains obtained after experimental passage through chickens. Concusions: All C. jejuni strains obtained from broiler chickens were typeable by the LG genotyping method. Application of the RsaI restriction enzyme improved the method in terms of ease...

  20. 空肠弯曲菌致病分子机制研究进展%Molecular Mechanisms on Pathogenesis of Campylobacter jejuni

    Institute of Scientific and Technical Information of China (English)

    李烨; 史锋; 王小元

    2009-01-01

    空肠弯曲菌(Campylobacter jejuni)是人类的食源性病原菌,感染后主要引起急性肠炎,与人类格林-巴利综合症也有密切关系.研究表明,空肠弯曲菌致病性是多种毒力因子共同作用的结果.作者从分子生物学研究水平综述空肠弯曲菌黏附、定植、侵入、产细胞毒素、分子模拟等致病机制.