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Sample records for acquired campylobacter jejuni

  1. Campylobacter jejuni organism (image)

    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 ...

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

    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.

  3. Compuesto bactericida contra Campylobacter jejuni

    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.

  4. 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

    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...... 1997 through 2007, C. jejuni isolates were obtained from The Danish Integrated Antimicrobial Resistance Monitoring and Research Programme (DANMAP) and susceptibility tested for ciprofloxacin, erythromycin, nalidixic acid, and tetracycline. Erythromycin resistance was at a low level in all the....... In Denmark, the use of fluoroquinolones in animal husbandry has been restricted since 2003. The purpose of the present study was to look at trends in occurrence of resistance among C. jejuni from broiler chickens, broiler chicken meat and human domestically acquired or travel associated cases. From...

  5. Media for Campylobacter jejuni and other campylobacters

    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

  6. Campylobacter jejuni in commercial eggs

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

    2014-01-01

    This study evaluated the ability of Campylobacter jejuni to penetrate through the pores of the shells of commercial eggs and colonize the interior of these eggs, which may become a risk factor for human infection. Furthermore, this study assessed the survival and viability of the bacteria in commercial eggs. The eggs were placed in contact with wood shavings infected with C. jejuni to check the passage of the bacteria. In parallel, the bacteria were inoculated directly into the air chamber to...

  7. Campylobacter jejuni acquire new host-derived CRISPR spacers when in association with bacteriophages harbouring a CRISPR-like Cas4 protein

    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. Improved Biotyping Schemes for Campylobacter jejuni and Campylobacter coli

    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.

  9. Survival of Campylobacter jejuni in Waterborne Protozoa

    Snelling, W. J.; McKenna, J. P.; Lecky, D. M.; Dooley, J. S. G.

    2005-01-01

    The failure to reduce the Campylobacter contamination of intensively reared poultry may be partially due to Campylobacter resisting disinfection in water after their internalization by waterborne protozoa. Campylobacter jejuni and a variety of waterborne protozoa, including ciliates, flagellates, and alveolates, were detected in the drinking water of intensively reared poultry by a combination of culture and molecular techniques. An in vitro assay showed that C. jejuni remained viable when in...

  10. Monoclonal Antibodies Specific for Hippurate Hydrolase of Campylobacter jejuni

    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.

  11. Oxygen requirement and tolerance of Campylobacter jejuni.

    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 ...

  12. Campylobacter jejuni--an emerging foodborne pathogen.

    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....

  13. Epidemiological aspects of Campylobacter jejuni enteritis.

    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...

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

    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.

  15. Septic abortion caused by Campylobacter jejuni bacteraemia.

    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

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

    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

  17. Virulence of Campylobacter jejuni for chicken embryos.

    Mahajan, S; Rodgers, F G

    1989-01-01

    The pathogenicity of Campylobacter jejuni was examined in chicken embryos. In this system, mortality data and histopathological findings induced by organisms and by bacterium-free filtered broth were identical. The absence in chicken embryo tissues both of organisms and of an inflammatory infiltrate suggests a toxin etiology.

  18. Campylobacter jejuni acquire new host-derived CRISPR spacers when in association with bacteriophages harboring a CRISPR-like Cas4 protein

    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...

  19. Genomic Characterization of Campylobacter jejuni strain M1

    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...

  20. Concerted evolution in Campylobacter jejuni and Campylobacter coli

    Concerted evolution is the phenomenon in which multiple copies of genes maintain sequence similarity in a single individual while the genes continue to diverge between individuals. Concerted evolution has been described in Campylobacter jejuni and C. coli for the pair of flagellin genes, which are ...

  1. Epidemiological aspects of Campylobacter jejuni enteritis.

    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

  2. Effect of gamma radiation on Campylobacter jejuni

    Radiation resistance of Campylobacter jejuni in broth, ground beef, and ground turkey meat was determined using dose levels from 0-200 Krad at -30 +/- 100C, at 0-50C, and at 30 +/- 100C. Irradiation at -300C 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 D10 value for C. jejuni was 32 Krad, which was less than D10 values commonly reported for salmonellae. 20 references, 4 figures

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

    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...

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

    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

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

    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...

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

    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).

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

    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

  8. Effect of gamma radiation on Campylobacter jejuni

    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.

  9. Substrate utilization by Campylobacter jejuni and Campylobacter coli

    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.

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

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

    1998-01-01

    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...

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

    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...

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

    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...

  13. Demonstration of Polysaccharide Capsule in Campylobacter jejuni Using Electron Microscopy

    Karlyshev, Andrey V.; McCrossan, Maria V.; Wren, Brendan W.

    2001-01-01

    Recently, we reported that Campylobacter jejuni, an important gastrointestinal pathogen, has the genetic determinants to produce a capsular polysaccharide (Karlyshev et al., Mol. Microbiol. 35:529–541, 2000). Despite these data, the presence of a capsule in these bacteria has remained controversial. In this study we stain C. jejuni cells with the cationic dye Alcian blue and demonstrate for the first time by electron microscopy that C. jejuni cells produce a polysaccharide capsule that is ret...

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

    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.

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

    Nielsen, Eva Møller; Nielsen, Niels Ladefoged

    1999-01-01

    Campylobacter infection is one of the most common bacterial enteric pathogens. Campylobacter jejuni and Campylobacter coli infections are mostly food- and waterborne and especially poultry is often assumed to be an important source. The heat-stable serotyping system (the 'Penner' scheme) was used...

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

    Horn Sharon T

    2007-05-01

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

  17. Cellular response of Campylobacter jejuni to trisodium phosphate

    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...

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

    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.

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

    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.

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

    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

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

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

    2008-01-01

    The vector potential of flies (Diptera: Brachycera) for spread of Campylobacter jejuni and Campylobacter coli on 5 Danish broiler farms was evaluated in a longitudinal field study from April to November 2004. First, the prevalence of C. jejuni- and C. coli-positive flies was determined in 2...... 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) was...... 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...

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

    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

  3. Prevalence of Campylobacter jejuni in poultry breeder flocks

    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.

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

    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...

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

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

    1998-01-01

    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....

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

    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...

  7. Survey of Campylobacter jejuni and Campylobacter coli in different taxa and ecological guilds of migratory birds

    Alessandro Fioretti

    2010-01-01

    Full Text Available A total of 169 faecal samples were collected from migrating birds, belonging to the Order of Passeriformes, in Campania region in order to isolate Campylobacter spp. Campylobacter spp. were isolated from 39 of the 169 birds examined (23.1%. Among these 36 were identified as C. jejuni and the remaining strains were identified as Campylobacter coli. Given the high isolation rates wild birds could be considered natural reservoir of infection.

  8. Metronidazole resistance in Campylobacter jejuni from poultry meat

    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...

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

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

    2012-01-01

    Dairy cows have been identified as common carriers of Campylobacter jejuni, which causes many of the human gastroenteritis cases reported worldwide. To design on-farm management practices that control the human infection sourced from dairy cows, the first step is to acquire an understanding of the excretion patterns of the cow reservoir. We monitored the same 35 cows from two dairy farms for C. jejuni excretion fortnightly for up to 12 months. The objective was to examine the concentration of...

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

    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.

  11. Colonization properties of Campylobacter jejuni in chickens

    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 ...

  12. Survival of Campylobacter jejuni in different gas mixtures

    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...

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

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

    2011-06-01

    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 genes were frequently present in both C. jejuni and C. coli isolated. Moreover, seven C. jejuni isolates carried also the wlaN gene which is presumably involved in the expression of ganglioside mimics in Guillain-Barré syndrome. PMID:21450419

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

    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

  15. Antimicrobial activities of isothiocyanates against Campylobacter jejuni isolates

    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.

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

    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...

  17. Metronidazole resistance in Campylobacter jejuni from poultry meat

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

    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...... 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 <0.005). This is the first report of foodborne bacteria carrying metronidazole resistance....

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

    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

  19. Detection and Typing of Campylobacter jejuni and Campylobacter coli and Analysis of Indicator Organisms in Three Waterborne Outbreaks in Finland

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

    2003-01-01

    Waterborne outbreaks associated with contamination of drinking water by Campylobacter jejuni are rather common in the Nordic countries Sweden, Norway, and Finland, where in sparsely populated districts groundwater is commonly used without disinfection. Campylobacters, Escherichia coli, or other coliforms have rarely been detected in potential sources. We studied three waterborne outbreaks in Finland caused by C. jejuni and used sample volumes of 4,000 to 20,000 ml for analysis of campylobacte...

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

    Phongsisay, Vongsavanh

    2016-04-01

    The Gram-negative bacterium Campylobacter jejuni causes gastroenteritis and Guillain-Barré syndrome in humans. Recent advances in the immunobiology of C. jejuni have been made. This review summarizes C. jejuni-binding innate receptors and highlights the role of innate immunity in autoimmune diseases. This human pathogen produces a variety of glycoconjugates, including human ganglioside-like determinants and multiple activators of Toll-like receptors (TLRs). Furthermore, C. jejuni targets MyD88, NLRP3 inflammasome, TIR-domain-containing adapter-inducing interferon-β (TRIF), sialic acid-binding immunoglobulin-like lectins (Siglecs), macrophage galactose-type lectin (MGL), and immunoglobulin-like receptors (TREM2, LMIR5/CD300b). The roles of these innate receptors and signaling molecules have been extensively studied. MyD88-mediated TLR activation or inflammasome-dependent IL-1β secretion is essential for autoimmune induction. TRIF mediates the production of type I interferons that promote humoral immune responses and immunoglobulin class-switching. Siglec-1 and Siglec-7 interact directly with gangliosides. Siglec-1 activation enhances phagocytosis and inflammatory responses. MGL internalizes GalNAc-containing glycoconjugates. TREM2 is well-known for its role in phagocytosis. LMIR5 recognizes C. jejuni components and endogenous sulfoglycolipids. Several lines of evidence from animal models of autoimmune diseases suggest that simultaneous activation of innate immunity in the presence of autoreactive lymphocytes or antigen mimicry may link C. jejuni to immunopathology. PMID:26709064

  1. A waterborne Campylobacter jejuni outbreak on a Greek island.

    Karagiannis, I; Sideroglou, T; Gkolfinopoulou, K; Tsouri, A; Lampousaki, D; Velonakis, E N; Scoulica, E V; Mellou, K; Panagiotopoulos, T; Bonovas, S

    2010-12-01

    A case-control and a case-crossover study were performed to investigate a Campylobacter jejuni outbreak in Crete in 2009. Most cases originated from rural areas, served by a different water-supply system from that of the adjacent town. Thirty-seven cases and 79 controls were interviewed; cases were interviewed for two different time periods for the case-crossover study. Stool cultures, PFGE and MLST subtyping were run in human samples. Univariately, consumption of tap water was associated with C. jejuni infection. Stratified analysis revealed that water-supply system was an effect modifier of this association. In the multivariable analysis, the rural areas' water supplier and drinking tap water were risk factors. No risk factors were revealed in the case-crossover study. No Campylobacter were isolated in the tested water samples. There is strong epidemiological evidence that tap water was the vehicle of the outbreak. PMID:20836911

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

    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,

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

    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...

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

    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.

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

    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...

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

    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...

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

    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, ...

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

    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...

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

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

    2015-01-01

    In this study we isolated novel bacteriophages, infecting the zoonotic bacterium Campylobacter jejuni. These phages may be used in phage therapy of C. jejuni colonized poultry to prevent spreading of the bacteria to meat products causing disease in humans. Many C. jejuni phages have been isolated...

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

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

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

    2008-01-01

    Responsible for the majority of bacterial gastroenteritis in the developed world, Campylobacter jejuni is a pervasive pathogen of humans and animals, but its evolution is obscure. In this paper, we exploit contemporary genetic diversity and empirical evidence to piece together the evolutionary history of C. jejuni and quantify its evolutionary potential. Our combined population genetics-phylogenetics approach reveals a surprising picture. Campylobacter jejuni is a rapidly evolving species, su...

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

    Humphrey, S; Chaloner, G; Kemmett, K; Davidson, N; Williams, N.; Kipar, A.; Humphrey, T.; Wigley, P.

    2014-01-01

    Campylobacter jejuni is the leading cause of bacterial food-borne infection; chicken meat is its main source. C. jejuni is considered commensal in chickens based on experimental models unrepresentative of commercial production. Here we show that the paradigm of Campylobacter commensalism in the chicken is flawed. Through experimental infection of four commercial breeds of broiler chickens, we show that breed has a significant effect on C. jejuni infection and the immune response of the animal...

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

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

    2012-01-01

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

  14. Role of Campylobacter jejuni gamma-glutamyl transpeptidase on epithelial cell apoptosis and lymphocyte proliferation

    Floch, Pauline; Pey, Vincent; Castroviejo, Michel; Dupuy, Jean William; Bonneu, Marc; de la Guardia, Anaïs Hocès; Pitard, Vincent; Mégraud, Francis; Lehours, Philippe

    2014-01-01

    Background A gamma-glutamyl transpeptidase (GGT) is produced by up to 31% of strains of Campylobacter jejuni isolates. C. jejuni GGT is close to Helicobacter pylori GGT suggesting a conserved activity but unlike the latter, C. jejuni GGT has not been studied extensively. In line with the data available for H. pylori, our objectives were to purify C. jejuni GGT from the bacteria, and to evaluate its inhibitory and proapoptotic activities on epithelial cells and human lymphocytes. Methods C. je...

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

    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

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

    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...

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

    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.

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

    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.

    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. Prevalence of Campylobacter jejuni and Campylobacter coli species in cats and dogs from Bydgoszcz (Poland) region.

    Andrzejewska, M; Szczepańska, B; Klawe, J J; Spica, D; Chudzińska, M

    2013-01-01

    The aim of this study was to investigate the role of cats and dogs as a potential reservoir of Campylobacter spp. Rectal swabs from 83 dogs and 71 cats were examined. Samples were obtained from the animals aged between 2 weeks and 24 months living in shelters, private households, farms and from veterinary clinics located in Bydgoszcz region during routine check-up. Campylobacter spp. were isolated from 4.81% dogs and 9.86% cats, respectively. C. jejuni was predominant in this study. All strains were isolated in autumn and winter from the animals living in farms and private houses. All the animals positive for Campylobacter prevalence had access to small water basins, accidental source of food and had contact with wild birds, poultry or their feaces. Isolates characterization revealed high prevalence of Campylobacter virulence genes-flaA, cadF and cdtB. 91% of isolated strains were susceptible to erythromycin. 81% among isolated strains were susceptible to azithromycin, 64% to tetracycline and 36% to ciprofloxacin. For 2 C. jejuni strains isolated from cats Random Amplified Polymorphic DNA (RAPD) profiling indicated 80% homology between them. PMID:23691584

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

    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.

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

    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

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

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

    2016-01-01

    In May 2014, the Utah Public Health Laboratory (UPHL) notified the Utah Department of Health (UDOH) of specimens from three patients infected with Campylobacter jejuni yielding indistinguishable pulsed-field gel electrophoresis (PFGE) patterns. All three patients had consumed raw (unpasteurized and nonhomogenized) milk from dairy A. In Utah, raw milk sales are legal from farm to consumer with a sales permit from the Utah Department of Agriculture and Food (UDAF). Raw milk dairies are required to submit monthly milk samples to UDAF for somatic cell and coliform counts, both of which are indicators of raw milk contamination. Before this cluster's identification, dairy A's routine test results were within acceptable levels (raw milk; the isolate matched the cluster pattern. UDAF suspended dairy A's raw milk permit during August 4-October 1, and reinstated the permit when follow-up cultures were negative. Additional cases of C. jejuni infection were identified in October, and UDAF permanently revoked dairy A's permit to sell raw milk on December 1. During May 9-November 6, 2014, a total of 99 cases of C. jejuni infection were identified. Routine somatic cell and coliform counts of raw milk do not ensure its safety. Consumers should be educated that raw milk might be unsafe even if it meets routine testing standards. PMID:27031585

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

    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.

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

    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...

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

    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...

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

    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

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

    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...

  9. Clonal population structure and antimicrobial resistance of Campylobacter jejuni from chicken meat in Belgium

    Campylobacter jejuni is one of the most important causes of human diarrhea worldwide. In the present work, multilocus sequence typing (MLST) was used to study the genotypic diversity of 145 C. jejuni isolates from 135 chicken meat preparations sampled across Belgium. Isolates were further typed by p...

  10. The microbiome structure and Campylobacter jejuni transcriptome in naturally-raised chickens

    Campylobacter jejuni is a leading cause of bacterially derived gastroenteritis in humans worldwide. C. jejuni regulates gene expression under various environmental conditions and stresses, indicative of its ability to survive in diverse niches. Few transcription factors have been identified, and the...

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

    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...

  12. Description of Campylobacter jejuni Bf, an atypical aero-tolerant strain

    Rodrigues, Ramila Christiane; Pocheron, Anne-Lise; Hernould, Michel; Haddad, Nabila; Tresse, Odile

    2015-01-01

    Background Campylobacter jejuni is a leading cause of bacterial enteritis worldwide. This microaerophilic bacterium can survive in aerobic environments, suggesting it has protective mechanisms against oxidative stress. The clinical C. jejuni Bf strain is characterized by an increased resistance to oxygen. This study aimed to characterize the behavior of the clinical C. jejuni Bf strain under an aerobic atmosphere and in response to ROS-promoter agents. Methods Growth was studied in both aerob...

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

    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...

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

    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...

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

    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 for...... strongly indicate that energy taxis is the primary force in environmental navigation by C. jejuni and that this mechanism drives the organism toward the optimal chemical conditions for energy generation and colonization....

  16. Basolateral Invasion and Trafficking of Campylobacter jejuni in Polarized Epithelial Cells

    Bouwman, L.I.; Niewold, P.; van Putten, J.P.M.

    2013-01-01

    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 ...

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

    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...

  18. Molecular Characterization of Invasive and Noninvasive Campylobacter jejuni and Campylobacter coli Isolates

    Carvalho, Alexandro C. T.; Ruiz-Palacios, Guillermo M.; Ramos-Cervantes, Pilar; Cervantes, Luz-Elena; Jiang, Xi; Pickering, Larry K.

    2001-01-01

    Campylobacter jejuni is one of the most common causes of bacterial diarrhea worldwide and is the primary bacterial cause of food-borne illness. Adherence to and invasion of epithelial cells are the most important pathogenic mechanisms of Campylobacter diarrhea. Molecular characterization of invasive and noninvasive Campylobacter isolates from children with diarrhea and symptom-free children was performed by random amplified polymorphic DNA techniques (RAPD). A distinct RAPD profile with a DNA band of 1.6 kb was observed significantly more frequently among invasive (63%) than among noninvasive (16%) Campylobacter isolates (P = 0.000005). The 1.6-kb band was named the invasion-associated marker (IAM). Using specifically designed primers, a fragment of 518 bp of the iam locus was amplified in 85% of invasive and 20% of noninvasive strains (P = 0.0000000). Molecular typing with a PCR-restriction fragment length polymorphism assay which amplified the entire iam locus showed a HindIII restriction fragment polymorphism pattern associated mainly with invasive strains. Although cluster analysis of the RAPD fingerprinting showed genetic diversity among strains, two main clusters were identified. Cluster I comprised significantly more pathogenic and invasive isolates, while cluster II grouped the majority of nonpathogenic, noninvasive isolates. These data indicate that most of the invasive Campylobacter strains could be differentiated from noninvasive isolates by RAPD analysis and PCR using specific primers that amplify a fragment of the iam locus. PMID:11283056

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

    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.

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

    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...

  1. Occurrence of Campylobacter jejuni and Giardia species in muskrat (Ondatra zibethica).

    Pacha, R E; Clark, G. W.; Williams, E A

    1985-01-01

    A total of 189 muskrat fecal samples were surveyed for Campylobacter and Giardia species. Campylobacter jejuni was recovered from 47.5% of these samples, and Giardia species were detected in 82.5%. These findings indicate that muskrat may be of importance to the health both of humans and of domestic animals.

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

    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...

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

    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...... specimens before slaughter is therefore vital to prevent disease transmission. By combining two techniques – immunomagnetic separation (IMS) and polymerase chain reaction (PCR), this study developed a reliable and specific method for rapid detection of C. jejuni in chicken fecal samples. The specificity 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...

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

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

    2001-01-01

    We investigated the genotype and serotype diversity of Campylobacter coli and C jejuni in two parent flocks of adult hens and their offspring over two rotations in order to evaluate the role of hatchery mediated transmission and/or vertical transmission of campylobacters in broiler flocks. In total......, 314 C jejuni and 32 C coli isolates from parent and broiler flocks and from the surroundings of broiler houses were typed by flagellin gene PCR/RFLP fla-typing), and selected isolates were also typed by serotyping and macrorestriction profiling using PFGE (MRP/PFGE). The combined typing results showed...... that the broiler flocks could be colonised by 1-3 different Campylobacter clones and parent flocks could be colonised by 2-6 different clones. C coli was isolated from up to 36% of birds in one parent flock, whereas only C jejuni was isolated from broiler flocks. C jejuni clones from different flocks...

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

    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.

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

    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.

  7. Tight junction changes in epithelial cells by Campylobacter jejuni and non-jejuni Campylobacter species

    Bücker, Roland; Nielsen, Hans Linde; Krüg, S;

    Campylobacter concisus infections of the gastrointestinal tract can be accompanied by diarrhea and inflammation, whereas colonization of the human oral cavity might have a commensal nature. We focus on the pathophysiology of C. concisus and the effects of different clinical oral and fecal C...

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

    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...

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

    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...

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

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

  11. Temperature-dependent phenotypic variation of Campylobacter jejuni lipooligosaccharides

    Moran Anthony P

    2010-11-01

    Full Text Available 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.

  12. Temperature-dependent phenotypic variation of Campylobacter jejuni lipooligosaccharides

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

    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.

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

    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

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

    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

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

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

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

    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 larvae to the adult fly. Conclusions The addition of fly larvae both accelerated the degradation of manure and...... C. jejuni. Pupae or newly hatched flies were not carriers of C. jejuni although larvae were grown in contaminated manure. Impact When composting poultry manure with Md fly larvae, it is possible both to reduce the amount of waste and to sanitize it from C. jejuni, thereby reducing the risk of...

  18. 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

    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

  19. Antimicrobial Resistance in Campylobacter jejuni and Campylobacter coli isolated from Chicken Carcass Rinsates: Update from the Animal Arm of NARMS

    Introduction: The development of antimicrobial resistance in Campylobacter species, particularly C. jejuni and C. coli, is of public health concern. Purpose: The purpose of this study was to analyze the minimum inhibitory concentrations (MICs) for antimicrobials used in susceptibility testing of C....

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

    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...

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

    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.

  2. Antimicrobial susceptibility of Campylobacter jejuni subsp. jejuni assessed by E-test and double dilution agar method in Southern Chile

    Fernández H

    2000-01-01

    Full Text Available The susceptibility patterns of 108 Campylobacter jejuni subsp. jejuni clinical strains, to six antimicrobial agents was determined by using the E-test and the double dilution agar methods. Using both metods, no strain was found to be resistant to ciprofloxacin, erythromycin and gentamicin, but two (1.8% were resistant to tetracycline and all to aztreonam. Seven (6.5% strains were resistant to ampicillin by the E-test and five (4.6% by the double dilution agar method and by both meyhods. No great discrepancies were observed between both methods.

  3. Exploring PFGE for Detecting Large Plasmids in Campylobacter jejuni and Campylobacter coli Isolated from Various Retail Meats

    Daya Marasini

    2014-10-01

    Full Text Available Campylobacter spp. is one of the most prevalent bacterial pathogens in retail meat, particularly poultry, and is a leading cause of diarrhea in humans. Studies related to Campylobacter large plasmids are limited in the literature possibly due to difficulty in isolating them using available alkaline lysis methods. The objectives of this study were to determine the prevalence of plasmids, particularly large ones, in Campylobacter spp. isolated from various Oklahoma retail meats, and to explore PFGE (Pulsed Field Gel Electrophoresis as a tool in facilitating the detection of these plasmids. One hundred and eighty nine strains (94 Campylobacter jejuni and 95 Campylobacter coli were screened for the presence of plasmids using both alkaline lysis and PFGE. Plasmids were detected in 119/189 (63% using both methods. Most of the plasmids detected by alkaline lysis were smaller than 90 kb and only three were larger than 90 kb. Plasmids over 70 kb in size were detected in 33 more strains by PFGE of which 11 strains contained larger than 90 kb plasmids. Plasmids were more prevalent in Campylobacter coli (73.5% than in Campylobacter jejuni (52%. BglII restriction analysis of plasmids isolated from 102 isolates revealed 42 different restriction patterns. In conclusion, PFGE was able to detect large plasmids up to 180 Kb in Campylobacter spp. which might have been missed if the alkaline lysis method was solely used. Campylobacter spp. isolated from retail meats harbor a diverse population of plasmids with variable sizes. To our knowledge, this is the first study to use PFGE to detect large plasmids in Campylobacter.

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

    Zeng, Ximin; Ardeshna, Devarshi; Lin, Jun

    2015-01-01

    Campylobacter jejuni, the leading bacterial cause of human gastroenteritis in the United States, displays significant strain diversity due to horizontal gene transfer. Conjugation is an important horizontal gene transfer mechanism contributing to the evolution of bacterial pathogenesis and antimicrobial resistance. It has been observed that heat shock could increase transformation efficiency in some bacteria. In this study, the effect of heat shock on C. jejuni conjugation efficiency and the ...

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

    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...

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

    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

  7. Rapid evolution and the importance of recombination to the gastroenteric pathogen Campylobacter jejuni.

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

    2009-02-01

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

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

    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....

  9. Western blot analysis of the human antibody response to Campylobacter jejuni cellular antigens during gastrointestinal infection.

    Nachamkin, I; Hart, A. M.

    1985-01-01

    Western blot analysis was used to identify antigenic components of Campylobacter jejuni whole cells and outer membranes that elicit antibody responses in patients with campylobacter enteritis. Acute- and convalescent-phase sera from eight patients were analyzed for antibody activity against their homologous infecting strains and heterologous clinical isolates. Whole-cell and Sarkosyl-insoluble membrane components were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and ...

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

    Oh, Euna; Jeon, Byeonghwa

    2015-01-01

    The increasing resistance of Campylobacter to clinically important antibiotics, such as fluoroquinolones and macrolides, is a serious public health problem. The objective of this study is to investigate synergistic anti-Campylobacter jejuni activity of fluoroquinolones and macrolides in combination with phenolic compounds. Synergistic antimicrobial activity was measured by performing a checkerboard assay with ciprofloxacin and erythromycin in the presence of 21 phenolic compounds. Membrane pe...

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

    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...

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

    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...

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

    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.

  14. Campylobacter jejuni strains of human and chicken origin are invasive in chickens after oral challenge

    Knudsen, Katrine Nørrelund; Bang, Dang Duong; Andresen, Lars Ole;

    2006-01-01

    The aim of the study was to evaluate the colonizing ability and the invasive capacity of selected Campylobacter jejuni strains of importance for the epidemiology of C jejuni in Danish broiler chickens. Four C jejuni strains were selected for experimental colonization Studies in day-old and 14-day......-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...

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

    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.

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

    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 and...... chemoreceptors have been predicted. In order to explore the role of chemotaxis in C. jejuni colonization we have constructed defined deletion mutants in the putative chemoreceptors. These mutants are analyzed for their motile characteristics and their chemotatic capacity in order to investigate the chemoreceptor...

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

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

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

    2003-03-01

    Waterborne outbreaks associated with contamination of drinking water by Campylobacter jejuni are rather common in the Nordic countries Sweden, Norway, and Finland, where in sparsely populated districts groundwater is commonly used without disinfection. Campylobacters, Escherichia coli, or other coliforms have rarely been detected in potential sources. We studied three waterborne outbreaks in Finland caused by C. jejuni and used sample volumes of 4,000 to 20,000 ml for analysis of campylobacters and sample volumes of 1 to 5,000 ml for analysis of coliforms and E. coli, depending on the sampling site. Multiple samples obtained from possible sources (water distribution systems and environmental water sources) and the use of large sample volumes (several liters) increased the chance of detecting the pathogen C. jejuni in water. Filtration of a large volume (1,000 to 2,000 ml) also increased the rate of detection of coliforms and E. coli. To confirm the association between drinking water contamination and illness, a combination of Penner serotyping and pulsed-field gel electrophoresis (digestion with SmaI and KpnI) was found to be useful. This combination reliably verified similarity or dissimilarity of C. jejuni isolates from patient samples, from drinking water, and from other environmental sources, thus confirming the likely reservoir of an outbreak. PMID:12620821

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

    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

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

    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.

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

    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

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

    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

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

    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

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

    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.

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

    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...

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

    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...

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

    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...

  8. The Seroprevalence of Campylobacter Jejuni in Patients with Irritable Bowel Syndrome and in Healthy Individuals

    Semnani, Sh. (MD

    2014-05-01

    Full Text Available Background and Objective: The patients with Post-Infectious Irritable bowel syndrome (PI-IBS, a subtype of irritable bowel syndrome, suffer from bacterial gastroenteritis. Since campylobacter Jejuni (CJ is one of the most common agents in this syndrome, we aimed to evaluate the prevalence of Campylobacter Jejuni in patients with Irritable Bowel Disease. Material and Methods: This case - control study was conducted on 160 patients divided into 2 equal groups of healthy and unhealthy. The presence of Anti- CJ antibody (IgG and IgA was evaluated by ELISA and the comparison was performed by chi-square test. Results: The mean age of case (31.51 and control (31.84 was not statistically different (P = 0.87. Titer of Anti- Campylobacter Jejuni antibody IgG was positive in 25% of patients and 18.8% of the healthy ones (p=0.02. IgA Seropositivity in patients was 7.5% but no one in control group was positive (p =0.01 Conclusion: The Seroprevalence of CJ in patients with IBS was higher significantly than that of control group. Thus, Cj can be known as one of the causes of Post-infection in patients with IBS in our region and it should be paid more attention in diagnostic assessment of these patients. Keyword: Irritable Bowel Syndrome; Campylobacter Jejuni; Antibody

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

    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...

  10. Comparative genomic analysis of clinical strains of Campylobacter jejuni from South Africa

    The bacterial foodborne pathogen Campylobacter jejuni is a common cause of acute gastroenteritis and is also associated with the postinfectious neuropathies, Guillain-Barré (GBS) and Miller Fisher (MFS) syndromes. This study described the use of multilocus sequence typing (MLST) and DNA microarrays ...

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

    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.

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

    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, was...... type (1/1) was represented by 44% of isolates, or by at least one isolate from 31 of 62 broiler flocks. This significantly exceeded the prevalence of fla type iii C. jejuni isolates that we have estimated from other studies and suggests that isolates carrying this fla type are overrepresented in Rocks...

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

    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.

  14. The role of probiotics in the inhibition of Campylobacter jejuni colonization and virulence attenuation.

    Mohan, V

    2015-08-01

    Campylobacter jejuni is one of the most common bacterial causes of human gastroenterocolitis worldwide, leading to diarrhea and other serious post-infectious complications. Probiotics form an attractive alternative intervention strategy for most of the enteric infections. However, the role of probiotics in C. jejuni infections requires detailed investigations in order to delineate the probiotic strains that are effective against C. jejuni. Although there are several biological mechanisms involved in the inhibition of pathogenic bacterial growth, the strains of probiotics and their mechanisms of actions through which they combat C. jejuni invasion have not been studied in greater detail. This mini review details the factors that are involved in the colonization and establishment of C. jejuni infection, with special reference to chickens, the natural host of C. jejuni, and the studies that have investigated the effect of different probiotic strains against C. jejuni colonization and growth. This review has collated the studies conducted using probiotics to inhibit C. jejuni colonization and growth to date to provide a collective knowledge about the role of probiotics as an alternative intervention strategy for campylobacteriosis. PMID:25934376

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

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

    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

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

    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.

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

    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

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

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

    2014-01-01

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

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

    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...

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

    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 to...... heat, starvation, oxidative or osmotic stresses encountered in the environment may affect the subsequent interaction of C. jejuni with free-living protozoa. To test this hypothesis, we examined the impact of environmental stress on expression of virulence-associated genes (ciaB, dnaJ, and htrA) of C...... could be recovered at this time point. Overall, pre-exposure of C. jejuni to environmental stresses did not promote intracellular survival in A. castellanii. Conclusions: Together, these findings suggest that the stress response in C. jejuni and its interaction with A. castellanii are complex and...

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

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

    2014-01-01

    Campylobacter jejuni is a major cause of bacterial gastroenteritis. C. jejuni encodes a protein glycosylation (Pgl) locus responsible for the N-glycosylation of membrane-associated proteins. We examined two variants of the genome sequenced strain NCTC11168: O, a representative of the original...

  3. Modification of the Campylobacter jejuni N-linked glycan by EptC protein-mediated addition of phosphoethanolamine

    Scott, Nichollas E; Nothaft, Harald; Edwards, Alistair V G;

    2012-01-01

    Campylobacter jejuni is the major worldwide cause of bacterial gastroenteritis. C. jejuni possesses an extensive repertoire of carbohydrate structures that decorate both protein and non-protein surface-exposed structures. An N-linked glycosylation system encoded by the pgl gene cluster mediates the...

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

    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

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

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

    2007-01-01

    The contribution of γ-glutamyl transpeptidase (GGT) to Campylobacter jejuni virulence and colonization of the avian gut has been investigated. The presence of the ggt gene in C. jejuni strains directly correlated with the expression of GGT activity as measured by cleavage and transfer of the γ-glutamyl moiety. Inactivation of the monocistronic ggt gene in C. jejuni strain 81116 resulted in isogenic mutants with undetectable GGT activity; nevertheless, these mutants grew normally in vitro. How...

  6. Altered synthetic response of Campylobacter jejuni to cocultivation with human epithelial cells is associated with enhanced internalization.

    Konkel, M E; Cieplak, W

    1992-01-01

    Campylobacter jejuni has been shown to bind to and enter epithelial cells in culture. The interaction of C. jejuni with INT 407 epithelial cells was examined to determine whether bacterial protein synthesis is required for either binding or internalization. Chloramphenicol, a selective inhibitor of bacterial protein synthesis, significantly reduced the internalization, but not binding, of C. jejuni compared with untreated controls as determined by protection from gentamicin. Electrophoretic a...

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

    Campylobacter jejuni is a leading cause of food- and waterbourne bacterial gastroenteritis in the developed world. Although illness is usually self-limiting, immunocompromised individuals are at risk for infections recalcitrant to antibiotic treatment, and prior campylobacter infection correlates wi...

  8. Closely related Campylobacter jejuni strains from different sources reveal a generalist rather than a specialist lifestyle

    Gripp Eugenia

    2011-11-01

    Full Text Available Abstract Background Campylobacter jejuni and Campylobacter coli are human intestinal pathogens of global importance. Zoonotic transmission from livestock animals or animal-derived food is the likely cause for most of these infections. However, little is known about their general and host-specific mechanisms of colonization, or virulence and pathogenicity factors. In certain hosts, Campylobacter species colonize persistently and do not cause disease, while they cause acute intestinal disease in humans. Results Here, we investigate putative host-specificity using phenotypic characterization and genome-wide analysis of genetically closely related C. jejuni strains from different sources. A collection of 473 fresh Campylobacter isolates from Germany was assembled between 2006 and 2010 and characterized using MLST. A subset of closely related C. jejuni strains of the highly prevalent sequence type ST-21 was selected from different hosts and isolation sources. PCR typing of strain-variable genes provided evidence that some genes differed between these strains. Furthermore, phenotypic variation of these strains was tested using the following criteria: metabolic variation, protein expression patterns, and eukaryotic cell interaction. The results demonstrated remarkable phenotypic diversity within the ST-21 group, which however did not correlate with isolation source. Whole genome sequencing was performed for five ST-21 strains from chicken, human, bovine, and food sources, in order to gain insight into ST-21 genome diversity. The comparisons showed extensive genomic diversity, primarily due to recombination and gain of phage-related genes. By contrast, no genomic features associated with isolation source or host were identified. Conclusions The genome information and phenotypic data obtained in vitro and in a chicken infection model provided little evidence of fixed adaptation to a specific host. Instead, the dominant C. jejuni ST-21 appeared to be

  9. Guillain-Barré Syndrome and Campylobacter jejuni Infection: A Review

    Nurun Nahar Mawla

    2014-01-01

    Full Text Available Guillain-Barré syndrome (GBS, a neurologic disease that produces ascending paralysis, affects people all over the world. Acute infectious illness precedes 50%-75% of the GBS cases. Although many infectious agents have been associated with GBS, the strongest documented association is with Campylobacter infection. The first line of evidence supporting Campylobacter infection as a trigger of GBS is anecdotal reports. The second line of evidence is serological surveys, which have demonstrated that sera from GBS patients contain anti Campylobacter jejuni antibodies, consistent with recent infection. Finally, culture studies have proven that a high proportion of GBS patients have C. jejuni in their stools at the time of onset of neurological symptoms. One of every 1058 Campylobacter infections results in GBS. Sialic acid containing lipooligosaccharides (LOS biosynthesis gene locus are associated with GBS and the expression of ganglioside mimicking structures. GM1a was the most prevalent ganglioside mimic in GBS associated strains. Molecular mimicry between C. jejuni LOS and gangliosides in human peripheral nerves, and cross-reactive serum antibody precipitate the majority of GBS cases in Bangladesh, like worldwide.

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

    Line, J Eric; Pearson, Kirsten G

    2003-10-01

    The purpose of these studies was to develop a conductimetric method for the rapid detection of Campylobacter jejuni. Numerous basal medium components were analyzed to develop a growth-enhancing broth medium for detection of freeze-injured Campylobacter cells using a conductimetric system. The final medium was composed of a modified Campy-Line agar from which the agar and triphenyltetrazolium chloride were removed and the amino acid, L-arginine was added. Pure isolates of C. jejuni. (frozen and thawed to produce stressed cells) were utilized to test the detection methodology. Monitoring of significant changes in the capacitance signal was found suitable for detection of Campylobacter proliferation. Using stressed pure cultures, Campylobacter growth was repeatedly detected at very low inoculum levels (about one cell per well). There was a direct linear relationship between detection times (DTs) and the initial inoculum level. For example, using a single strain, the mean DT (n = 20) at the 10 CFU/ml inoculum level was 28.6 h, with 100% of the inoculated wells detecting. The mean DTs at the 100, 1,000, and 10,000 CFU/ml inoculum levels were 24.9, 21.4, and 17.0 h, respectively. This study demonstrates that conductimetric methods can be utilized for the rapid detection of C. jejuni. PMID:14572208

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

    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.

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

    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...

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

    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.

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

    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

  15. Survival at refrigeration and freezing temperatures of Campylobacter coli and Campylobacter jejuni on chicken skin applied as axenic and mixed inoculums.

    El-Shibiny, Ayman; Connerton, Phillippa; Connerton, Ian

    2009-05-31

    Campylobacter is considered to be the most common cause of bacterial diarrhoeal illness in the developed world. Many cases are thought to be acquired from consumption of undercooked poultry. The aim of this study was to compare the effect of the rate of cooling on the survival, at 4 degrees C and -20 degrees C, of Campylobacter coli and Campylobacter jejuni strains, inoculated on chicken skin from axenic culture or as mixed inoculums. Strains chilled in a domestic refrigerator varied in their tolerance to storage at 4 degrees C. Statistically significant differences between strains applied as axenic or mixed inoculums were observed for specific strain combinations using two-way ANOVA, including the enhanced survival of antibiotic resistant C. coli 99/367 at 4 degrees C. The use of rapid cooling (at -20 degrees C/min) enhanced the survival of all the Campylobacter strains chilled to 4 degrees C compared to standard refrigeration. Freezing to -20 degrees C reduced viable counts by 2.2-2.6 log10 CFU/cm(2) in 24 h. Rapid cooling to -20 degrees C (at -30 degrees C/min) enhanced the survival of C. coli 99/367 compared to freezing in a domestic freezer. Statistically significant interaction terms between specific strains were observed in mixed inoculums chilled to -20 degrees C by freezing in a domestic freezer and by rapid chilling to -20 degrees C. Rapid chilling of poultry, particularly for 4 degrees C storage may enhance survival of Campylobacter and although this is an issue that affects meat quality, it should be considered by poultry processors. PMID:19324444

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

    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.

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

    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.

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

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

    2014-08-01

    Campylobacter jejuni is the most common cause of bacterial gastroenteritis in humans. The synthesis of cytolethal distending toxin appears essential in the infection process. In this work we evaluated the sequence of lethal events in HeLa cells exposed to cell lysates of two distinct strains, C. jejuni ATCC 33291 and C. jejuni ISS3. C. jejuni cell lysates (CCLys) were added to HeLa cell monolayers which were analysed to detect DNA content, death features, bcl-2 and p53 status, mitochondria/lysosomes network and finally, CD54 and CD59 alterations, compared to cell lysates of C. jejuni 11168H cdtA mutant. We found mitochondria and lysosomes differently targeted by these bacterial lysates. Death, consistent with apoptosis for C. jejuni ATCC 33291 lysate, occurred in a slow way (>48 h); concomitantly HeLa cells increase their endolysosomal compartment, as a consequence of toxin internalization besides a simultaneous and partial lysosomal destabilization. C. jejuni CCLys induces death in HeLa cells mainly via a caspase-dependent mechanism although a p53 lysosomal pathway (also caspase-independent) seems to appear in addition. In C. jejuni ISS3-treated cells, the p53-mediated oxidative degradation of mitochondrial components seems to be lost, inducing the deepest lysosomal alterations. Furthermore, CD59 considerably decreases, suggesting both a degradation or internalisation pathway. CCLys-treated HeLa cells increase CD54 expression on their surface, because of the action of lysate as its double feature of toxin and bacterial peptide. In conclusion, we revealed that C. jejuni CCLys-treated HeLa cells displayed different features, depending on the particular strain. PMID:24880782

  19. Identification of Multiple Subtypes of Campylobacter jejuni in Chicken Meat and the Impact on Source Attribution

    John A. Hudson

    2013-09-01

    Full Text Available Most source attribution studies for Campylobacter use subtyping data based on single isolates from foods and environmental sources in an attempt to draw epidemiological inferences. It has been suggested that subtyping only one Campylobacter isolate per chicken carcass incurs a risk of failing to recognise the presence of clinically relevant, but numerically infrequent, subtypes. To investigate this, between 21 and 25 Campylobacter jejuni isolates from each of ten retail chicken carcasses were subtyped by pulsed-field gel electrophoresis (PFGE using the two restriction enzymes SmaI and KpnI. Among the 227 isolates, thirteen subtypes were identified, the most frequently occurring subtype being isolated from three carcasses. Six carcasses carried a single subtype, three carcasses carried two subtypes each and one carcass carried three subtypes. Some subtypes carried by an individual carcass were shown to be potentially clonally related. Comparison of C. jejuni subtypes from chickens with isolate subtypes from human clinical cases (n = 1248 revealed seven of the thirteen chicken subtypes were indistinguishable from human cases. None of the numerically minor chicken subtypes were identified in the human data. Therefore, typing only one Campylobacter isolate from individual chicken carcasses may be adequate to inform Campylobacter source attribution.

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

    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....

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

    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.

  2. Altered synthetic response of Campylobacter jejuni to cocultivation with human epithelial cells is associated with enhanced internalization.

    Konkel, M E; Cieplak, W

    1992-01-01

    Campylobacter jejuni has been shown to bind to and enter epithelial cells in culture. The interaction of C. jejuni with INT 407 epithelial cells was examined to determine whether bacterial protein synthesis is required for either binding or internalization. Chloramphenicol, a selective inhibitor of bacterial protein synthesis, significantly reduced the internalization, but not binding, of C. jejuni compared with untreated controls as determined by protection from gentamicin. Electrophoretic analysis of metabolically labeled proteins revealed that C. jejuni cultured with INT 407 cells synthesized 14 proteins that were not detected in organisms cultured in medium alone. The inhibitory effect of chloramphenicol on internalization was reduced by preincubation of C. jejuni with INT 407 cells. The results indicate that C. jejuni, like some other enteric pathogens, engages in a directed response to cocultivation with epithelial cells by synthesizing one or more proteins that facilitate internalization and suggest that this phenomenon is relevant to the pathogenesis of enteritis caused by C. jejuni. Images PMID:1399005

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

    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...

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

    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.

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

    Anderson, Robin C; Vodovnik, Maša; Min, Byeng R; Pinchak, William E; Krueger, Nathan A; Harvey, Roger B; Nisbet, David J

    2012-07-01

    Strategies are sought to reduce intestinal colonisation of food-producing animals by Campylobacter jejuni, a leading bacterial cause of human foodborne illness worldwide. Presently, we tested the antimicrobial activity of hydrolysable-rich blackberry, cranberry and chestnut tannin extracts and condensed tannin-rich mimosa, quebracho and sorghum tannins (each at 100 mg/mL) against C. jejuni via disc diffusion assay in the presence of supplemental casamino acids. We found that when compared to non-tannin-treated controls, all tested tannins inhibited the growth of C. jejuni and that inhibition by the condensed tannin-rich mimosa and quebracho extracts was mitigated in nutrient-limited medium supplemented with casamino acids. When tested in broth culture, both chestnut and mimosa extracts inhibited growth of C. jejuni and this inhibition was much greater in nutrient-limited than in full-strength medium. Consistent with observations from the disc diffusion assay, the inhibitory activity of the condensed tannin-rich mimosa extracts but not the hydrolysable tannin-rich chestnut extracts was mitigated by casamino acid supplementation to the nutrient-limited medium, likely because the added amino acids saturated the binding potential of the condensed tannins. These results demonstrate the antimicrobial activity of various hydrolysable and condensed tannin-rich extracts against C. jejuni and reveal that condensed tannins may be less efficient than hydrolysable tannins in controlling C. jejuni in gut environments containing high concentrations of amino acids and soluble proteins. PMID:22528299

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

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

    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

  8. Chemical decontamination of Campylobacter jejuni on chicken skin and meat

    Nielsen, C. T.; Brondsted, L.O.; Rosenquist, Hanne;

    2007-01-01

    Campylobacter is a leading course of food borne bacterial infections worldwide. It is believed that a decline in the occurrence of campylobacteriosis can be achieved by reducing the number of the bacterium in the gastrointestinal tract of poultry. Phage therapy using Campylobacter specific phages...... given to poultry prior to slaughter is a promising control measure. However, the reducing effect of most phages tested so far is rather limited due to the development of phage resistant Campylobacter. An increased knowledge of the molecular mechanisms of importance for development of phage resistant...... Campylobacter is therefore essential for implementation of this method. Recently, a collection of Campylobacter bacteriophages has been established. The phages were isolated from faeces from broilers and ducks and it was shown by transmission electron microscopy that they all belong to the family of Myoviridae...

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

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

    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 ef....... Some of these DGGE bands could be affiliated with Lactobacillus reuteri, Clostridium perfringens, and the genus Klebsiella....

  10. High-voltage electroporation of bacteria: genetic transformation of Campylobacter jejuni with plasmid DNA.

    Miller, J F; Dower, W J; Tompkins, L S

    1988-01-01

    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...

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

    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...

  12. The Natural Antimicrobial Carvacrol Inhibits Campylobacter jejuni Motility and Infection of Epithelial Cells

    van Alphen, L; Burt, S. A.; Veenendaal, A.K.J.; Bleumink-Pluym, N; van Putten, J.P.M.

    2012-01-01

    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. ...

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

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

    2015-01-01

    Biofilms make an important contribution to survival and transmission of bacterial pathogens in the food chain. The human pathogen Campylobacter jejuni is known to form biofilms in vitro in food chain-relevant conditions, but the exact roles and composition of the extracellular matrix are still not clear. Extracellular DNA has been found in many bacterial biofilms and can be a major component of the extracellular matrix. Here we show that extracellular DNA is also an important component of the...

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

    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...

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

    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...

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

    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,

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

    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.

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

    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.

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

    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. PMID:25002548

  20. Performance of a 70-mer oligonucleotide microarray for genotyping of Campylobacter jejuni

    Ljungström Marianne

    2008-05-01

    Full Text Available Abstract Background Campylobacter jejuni is widespread in the environment and is the major cause of bacterial gastroenteritis in humans. In the present study we use microarray-based comparative genomic hybridizations (CGH, pulsed-field gel electrophoresis (PFGE and multilocus sequence typing (MLST to analyze closely related C. jejuni isolates from chicken and human infection. Results With the exception of one isolate, the microarray data clusters the isolates according to the five groups determined by PFGE. In contrast, MLST defines only three genotypes among the isolates, indicating a lower resolution. All methods show that there is no inherit difference between isolates infecting humans and chicken, suggesting a common underlying population of C. jejuni. We further identify regions that frequently differ between isolates, including both previously described and novel regions. Finally, we show that genes that belong to certain functional groups differ between isolates more often than expected by chance. Conclusion In this study we demonstrated the utility of 70-mer oligonucleotide microarrays for genotyping of Campylobacter jejuni isolates, with resolution outperforming MLST.

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

    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.

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

    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.

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

    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.

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

    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.

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

    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.

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

    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.

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

    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

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

    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

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

    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.

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

    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

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

    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.

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

    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

    2008-04-01

    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%), virB11 (0%), ceuE (82.5%), cdtA (97.5%), cdtB (97.5%), cdtC (97.5%), and wlaN (7.5%) genes was detected in C. jejuni by PCR. All C. jejuni strains but one produced cytolethal distending toxin in a HeLa cell assay. PMID:18287317

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

    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

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

    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 ...

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

    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...

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

    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 ...

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

    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 ...

  18. Contribution of TAT System Translocated PhoX to Campylobacter jejuni Phosphate Metabolism and Resilience to Environmental Stresses

    Drozd, Mary; Gangaiah, Dharanesh; Liu, Zhe; Rajashekara, Gireesh

    2011-01-01

    Campylobacter jejuni is a common gastrointestinal pathogen that colonizes food animals; it is transmitted via fecal contamination of food, and infections in immune-compromised people are more likely to result in serious long-term illness. Environmental phosphate is likely an important sensor of environmental fitness and the ability to obtain extracellular phosphate is central to the bacteria's core metabolic responses. PhoX is the sole alkaline phosphatase in C. jejuni, a substrate of the TAT...

  19. Metabolomic analysis of the food-borne pathogen Campylobacter jejuni: application of direct injection mass spectrometry for mutant characterisation

    Howlett, R.M; Kelly, D. J.; Davey, M P; Paul Quick, W

    2014-01-01

    Campylobacter jejuni is the most frequent cause of human food-borne bacterial gastroenteritis but its physiology and biochemistry are poorly understood. Only a few amino-acids can be catabolised and these are known to be important for host colonization. Here we have established methods for rapid high throughput analyses of global metabolism in C. jejuni using direct injection mass spectrometry (DIMS) to compare metabolite fingerprints of wild-type and mutant strains. Principal component analy...

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

    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...

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

    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...

  2. Large-Scale Comparative Genomics Meta-Analysis of Campylobacter jejuni Isolates Reveals Low Level of Genome Plasticity

    Taboada, Eduardo N.; Acedillo, Rey R; Carrillo, Catherine D.; Findlay, Wendy A.; Medeiros, Diane T.; Mykytczuk, Oksana L; Roberts, Michael J.; Valencia, C. Alexander; Farber, Jeffrey M.; Nash, John H E

    2004-01-01

    We have used comparative genomic hybridization (CGH) on a full-genome Campylobacter jejuni microarray to examine genome-wide gene conservation patterns among 51 strains isolated from food and clinical sources. These data have been integrated with data from three previous C. jejuni CGH studies to perform a meta-analysis that included 97 strains from the four separate data sets. Although many genes were found to be divergent across multiple strains (n = 350), many genes (n = 249) were uniquely ...

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

    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

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

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

    2016-01-01

    In this study, we investigated the distribution, phenotypic and molecular typing characters of Campylobacter jejuni in domestic fowl, and livestock populations in East China, to provide some reference for researches on its molecular epidemiology. A total of 1250 samples were collected from different animal sources, and C. jejuni strains were then isolated and tested for antibiotic sensitivity. Antibiotics-resistance gene and pathogenic genes were detected by polymerase chain reaction. Phylogenic analysis on the C. jejuni strains was performed by multilocus sequence typing (MLST) method. The results showed that 108 out of the 1250 samples (mean 8.64%) were C. jejuni positive. These 108 C. jejuni strains were highly sensitive to antibiotics such as chloramphenicol, amoxicillin, amikacin, cefotaxime, and azithromycin, whereas they were highly resistant to antibiotics such as cefoperazone, cotrimoxazole, cefamandole, sulfamethoxazole, and cefradine. Pathogenicity related gene identification indicated that the mean carrying rate of adhesion related gene cadF and racR, flagellin gene flaA, toxin regulating gene cdtA, cdtB, cdtC, wlaN and virB11, heat shock proteins and transferring proteins related genes dnaJ and ceuE, CiaB and pldA were 92.45%, 38.69%, 73.58%, 71.70%, 52.83%, 96.23%, 12.26%, 1.89%, 0.94%, 65.09%, 39.62% and 9.43%, respectively. A total of 58.82% of these strains contained more than 6 pathogenicity-related genes. MLST typed 58 ST types from the 108 isolated C. jejuni strains, including 24 new types, and ST-21 was the major type, accounting for 39.3% of the total strains. PMID:26565657

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

    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

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

    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

  7. The biofilm matrix of Campylobacter jejuni determined by fluorescence lectin-binding analysis.

    Turonova, Hana; Neu, Thomas R; Ulbrich, Pavel; Pazlarova, Jarmila; Tresse, Odile

    2016-05-01

    Campylobacter jejuni is responsible for the most common bacterial foodborne gastroenteritis. Despite its fastidious growth, it can survive harsh conditions through biofilm formation. In this work, fluorescence lectin-binding analysis was used to determine the glycoconjugates present in the biofilm matrix of two well-described strains. Screening of 72 lectins revealed strain-specific patterns with six lectins interacting with the biofilm matrix of both strains. The most common sugar moiety contained galactose and N-acetylgalactosamine. Several lectins interacted with N-acetylglucosamine and sialic acid, probably originated from the capsular polysaccharides, lipooligosaccharides and N-glycans of C. jejuni. In addition, glycoconjugates containing mannose and fucose were detected within the biofilm, which have not previously been found in the C. jejuni envelope. Detection of thioflavin T and curcumin highlighted the presence of amyloids in the cell envelope without association with specific cell appendages. The lectins ECA, GS-I, HMA and LEA constitute a reliable cocktail to detect the biofilm matrix of C. jejuni. PMID:27097059

  8. Partial Proteome Map of Campylobacter Jejuni Strain Nctc11168 by Gel-Free Proteomics Analysis

    Zilun Shi

    2014-12-01

    Full Text Available A proteome map of the foodborne pathogen Campylobacter jejuni NCTC11168 was analyzed using a state-of-the-art gel-free proteomic approach for the first time. A whole cell protein extract was prepared from the C. jejuni strain NCTC11168 grown in brain heart infusion (BHI broth at 42°C under microaerobic conditions. A gel-free technique using isobaric tags for relative and absolute quantitation (iTRAQ was employed to create a protein expression profile of the strain. Liquid chromatography-mass spectrometry (LC-MS/MS was used to identify the proteins. Protein functionalities were searched to classify them. A total of 235 proteins were identified in the whole cell protein fraction of C. jejuni NCTC11168 cells using iTRAQ analysis. Functional grouping of the identified proteins showed that forty percent of these proteins were associated with energy metabolism, protein synthesis and genetic information processing. iTRAQ was faster, easier and proved more sensitive than two-dimensional gel-based proteomics approaches previously applied to C. jejuni, making it an attractive tool for further studies of cellular physiological response.

  9. Comparison of virulence-associated in vitro properties of typed strains of Campylobacter jejuni from different sources

    Coote, J.G.; Stewart-Tull, D.E.S.; Owen, R.J.;

    2007-01-01

    Campylobacter jejuni is a major cause of human diarrhoeal disease, but specific virulence mechanisms have not been well defined. This blinded study was undertaken with 40 C. jejuni isolates from different sources to determine their haemolytic, cytotoxic and adhesion and invasion activities towards...... correlation between cytotoxicity and cell invasion was evident. Five AFLP clusters, each representing two to eleven isolates with similar profiles, were observed at the 90% similarity level. Some AFLP groups contained isolates with a common serotype, but each group had C. jejuni isolates from more than one...

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

    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

  11. 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

    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...

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

    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....... Interestingly, the dissolved oxygen levels of co-cultures with or without amoebae – bacteria contact were much lower than those observed with culture media or with C. jejuni alone incubated in aerobic conditions, and were comparable with levels obtained after 24 h of growth of C. jejuni under microaerophilic...

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

    Rastyani; Alikhani; Sedighi; Kazemi; Farhadi Kohan; Arabestani

    2015-01-01

    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. ...

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

    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.

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

    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...

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

    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...

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

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

    2014-09-01

    This study aimed to investigate the prevalence of Campylobacter jejuni in potential contamination sources that are not regularly monitored such as free-living urban pigeons and crows, dogs, cats and urban environmental water and to assess the possible impact on the epidemiology of campylobacteriosis in children using multilocus sequence typing (MLST). Campylobacter spp. were detected in 36.2 % of faecal samples of free-living urban birds and in 40.4 % of environmental water samples. A low prevalence of Campylobacter spp. was detected in dogs and cats, with 7.9 and 9.1 %, respectively. Further identification of isolates revealed that environmental water and pet samples were mostly contaminated by other Campylobacter spp. than C. jejuni, whereas C. jejuni was the most prevalent species in faecal samples of free-living birds (35.4 %). This species was the dominant cause of campylobacteriosis in children (91.5 %). In addition, the diversity of C. jejuni MLST types in free-living birds and children was investigated. Clonal complex (CC) 179 was predominant among free-living urban birds; however, only two isolates from children were assigned to this CC. One dog and one child isolate were assigned to the same clonal complex (CC48) and sequence type (ST) 918. The dominant two clonal complexes among the child clinical isolates (CC353 and CC21) were not detected among C. jejuni strains isolated from environmental sources examined in this study. As only two CCs were shared by environmental and child C. jejuni isolates and a high number of novel alleles and STs were found in C. jejuni isolated from free-living urban birds and environmental water, there is probably only a limited link between urban environmental sources and campylobacteriosis in children, particularly in rather cold climatic conditions. PMID:24987101

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

    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

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

    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.

  20. 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.

    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

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

    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.

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

    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

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

    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)

  4. Draft Genome Sequence of Lactobacillus crispatus JCM5810, Which Can Reduce Campylobacter jejuni Colonization in Chicken Intestine

    Wooten, Jessica; Liu, Xiaoji; Miller, Michael J.

    2016-01-01

    We present the 2.05-Mb draft genome sequence of Lactobacillus crispatus JCM5810, a chicken intestinal isolate with the ability to reduce Campylobacter jejuni colonization in chickens. The genome sequence will provide insights on the probiotic mechanisms of L. crispatus JCM5810.

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

    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

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

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

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

    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.

  8. Draft Genome Sequence of Lactobacillus crispatus JCM5810, Which Can Reduce Campylobacter jejuni Colonization in Chicken Intestine

    Wooten, Jessica; Liu, Xiaoji

    2016-01-01

    We present the 2.05-Mb draft genome sequence of Lactobacillus crispatus JCM5810, a chicken intestinal isolate with the ability to reduce Campylobacter jejuni colonization in chickens. The genome sequence will provide insights on the probiotic mechanisms of L. crispatus JCM5810. PMID:27081134

  9. The in vivo efficacy of two administration routes of a phage cocktail to reduce numbers of Campylobacter coli and Campylobacter jejuni in chickens

    Carvalho Carla M

    2010-09-01

    Full Text Available Abstract Background Poultry meat is one of the most important sources of human campylobacteriosis, an acute bacterial enteritis which is a major problem worldwide. Campylobacter coli and Campylobacter jejuni are the most common Campylobacter species associated with this disease. These pathogens live in the intestinal tract of most avian species and under commercial conditions they spread rapidly to infect a high proportion of the flock, which makes their treatment and prevention very difficult. Bacteriophages (phages are naturally occurring predators of bacteria with high specificity and also the capacity to evolve to overcome bacterial resistance. Therefore phage therapy is a promising alternative to antibiotics in animal production. This study tested the efficacy of a phage cocktail composed of three phages for the control of poultry infected with C. coli and C. jejuni. Moreover, it evaluated the effectiveness of two routes of phage administration (by oral gavage and in feed in order to provide additional information regarding their future use in a poultry unit. Results The results indicate that experimental colonisation of chicks was successful and that the birds showed no signs of disease even at the highest dose of Campylobacter administered. The phage cocktail was able to reduce the titre of both C. coli and C. jejuni in faeces by approximately 2 log10 cfu/g when administered by oral gavage and in feed. This reduction persisted throughout the experimental period and neither pathogen regained their former numbers. The reduction in Campylobacter titre was achieved earlier (2 days post-phage administration when the phage cocktail was incorporated in the birds' feed. Campylobacter strains resistant to phage infection were recovered from phage-treated chickens at a frequency of 13%. These resistant phenotypes did not exhibit a reduced ability to colonize the chicken guts and did not revert to sensitive types. Conclusions Our findings provide

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

    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.

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

    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

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

    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

  13. Hydrophobicity and aggregation of Campylobacter jejuni, Escherichia coli 0157:H7 and Listeria monocytogenes

    Trachoo, N.

    2005-03-01

    Full Text Available Physicochemical properties such as hydrophobicity and ability to aggregate play an important role in the attachment of microorganisms to environmental surfaces and intestinal tissues of humans and animals leading to gastroenteritis in hosts. The objective of this research was to determine relative hydrophobicity and aggregation properties of major foodborne pathogens including Campylobacter jejuni, Escherichia coli 0157:H7 and Listeria monocytogenes LM18, LM32, LM msu1 and LM msu2. Relative hydrophobicity as determined by the microbial adhesion to hydrocarbon (MATH test of E. coli 0157:H7 was 19.30% indicating that substances on its cell surfaces were more hydrophobic than other pathogens and thus could be extracted more by the hydrocarbon, hexadecane. C. jejuni had relative hydrophobicity of 4.15% while the four strains of L. monocytogenes had relative hydrophobicity ranging from 3.79% to 16.23%. Aggregation of C. jejuni, E. coli and L. monocytogenes LM18, LM32, LM msu1 and LM msu2 were 45%, 21.25%, 12.51%, 13.09%, 31.27% and 41.22%, respectively.

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

    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.

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

    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.

  16. Plant-derived compounds inactivate antibiotic-resistant Campylobacter jejuni strains.

    Ravishankar, Sadhana; Zhu, Libin; Law, Bibiana; Joens, Lynn; Friedman, Mendel

    2008-06-01

    Sixty-three Campylobacter jejuni isolates were screened for their resistance to the antibiotics ampicillin, cefaclor, ciprofloxacin, erythromycin, gentamycin, tetracycline, and trimethoprim-sulfamethoxazole. Based on this screen, the resistant strains D28a and H2a and the nonresistant strain A24a were selected for evaluation of their resistance and susceptibility to inactivation by cinnamaldehyde and carvacrol, the main constituents of plant-derived cinnamon and oregano oils, respectively. Different concentrations (0.05, 0.1, and 0.2% [vol/vol] in sterile phosphate-buffered saline) of cinnamaldehyde and carvacrol were added to C. jejuni cultures with initial populations of 10(4) CFU/ml. The samples were then mixed thoroughly and incubated at 37 degrees C. Viable bacterial populations were enumerated at incubation periods of 0, 30, 60, and 120 min. The results indicate that the extent of inhibition of microbial survival was related to both the nature and concentration of antimicrobials and the incubation time. Both cinnamaldehyde and carvacrol exhibited rapid antimicrobial activity against both antibiotic-resistant and non-resistant C. jejuni strains, at concentrations of approximately 0.1% and higher. The antimicrobial efficacy of cinnamaldehyde was greater than that of carvacrol. The possible significance of the results for microbiological food safety is discussed. PMID:18592739

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

    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. Antimicrobial resistance profiles and mechanisms of resistance in Campylobacter jejuni isolates from pets.

    Acke, Els; McGill, Kevina; Quinn, Teresa; Jones, Boyd R; Fanning, Seamus; Whyte, Paul

    2009-01-01

    The presence of antimicrobial resistance in 51 Campylobacter jejuni isolates obtained from cats and dogs was determined by E-testing. Resistance to nalidixic acid (37.3% of isolates), ciprofloxacin (19.6%), tetracycline (13.7%), ampicillin (13.7%), erythromycin (11.8%), and chloramphenicol (5.9%) was detected. Resistance to two antimicrobials or more was present in 31.4% of isolates, and one isolate was resistant to all six antimicrobials. Of the isolates with ciprofloxacin and/or nalidixic acid resistance, 54.5% had the gyrA substitution Thr-86-Ile on sequencing. The tet o gene was detected in 75.0% isolates with high-level resistance to tetracycline. With the observed antimicrobial resistance in C. jejuni isolates from pets in this study, and the detection of identical mechanisms for quinolone and tetracycline resistance in pets and humans, pets should be considered a potential source of (multi)resistant C. jejuni infections in humans. PMID:19580444

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

    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

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

    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

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

    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....

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

    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...

  3. Seroprevalence in Chickens against Campylobacter jejuni Flagellar Capping Protein (FliD) in Selected Areas of the United States.

    Yeh, H-Y; Hiett, K L; Line, J E; Jagne, J F; Lauer, D C

    2016-06-01

    Campylobacter jejuni is a causative pathogen of human acute bacterial gastroenteritis. Infected poultry products are regarded as a major source for human C. jejuni infection. The flagellar capping protein (FliD) is highly conserved among C. jejuni strains/isolates and is antigenic as analysed by immunoblot. In this study, we used the FliD protein as a probe to survey the prevalence of C. jejuni antibodies in chickens from two areas in the United States. A total of 394 samples were tested. Sera from layer breeders of 44-52 weeks of age tested 100% positive, while 4- to 6-week broilers from 22 premises showed 7-100% positivity. These results demonstrate that anti-FliD antibodies were prevalent in the poultry population in the areas of serum samples collected. PMID:26603949

  4. Pancreatic amylase is an environmental signal for regulation of biofilm formation and host interaction in Campylobacter jejuni.

    Jowiya, Waheed; Brunner, Katja; Abouelhadid, Sherif; Hussain, Haitham A; Nair, Sean P; Sadiq, Sohaib; Williams, Lisa K; Trantham, Emma K; Stephenson, Holly; Wren, Brendan W; Bajaj-Elliott, Mona; Cogan, Tristan A; Laws, Andrew P; Wade, Jim; Dorrell, Nick; Allan, Elaine

    2015-12-01

    Campylobacter jejuni is a commensal bacterium in the intestines of animals and birds and a major cause of food-borne gastroenteritis in humans worldwide. Here we show that exposure to pancreatic amylase leads to secretion of an α-dextran by C. jejuni and that a secreted protease, Cj0511, is required. Exposure of C. jejuni to pancreatic amylase promotes biofilm formation in vitro, increases interaction with human epithelial cell lines, increases virulence in the Galleria mellonella infection model, and promotes colonization of the chicken ileum. We also show that exposure to pancreatic amylase protects C. jejuni from stress conditions in vitro, suggesting that the induced α-dextran may be important during transmission between hosts. This is the first evidence that pancreatic amylase functions as an interkingdom signal in an enteric microorganism. PMID:26438798

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

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

    2016-01-01

    birds eating food of animal or mixed animal and vegetable origin and foraging on the ground close to livestock were more likely to carry Campylobacter spp. than those foraging further away or hunting in the air. These findings suggest that wild birds may play a role in sustaining the epidemiology of...... general, birds feeding on a diet of animal or mixed animal and vegetable origin, foraging on the ground and vegetation in close proximity to livestock stables were more likely to carry Campylobacter spp. in both summer (P <0.001) and winter (P <0.001) than birds foraging further away from the farm or in...

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

    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...

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

    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

  8. 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

    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 of...

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

    Teh, Amy Huei Teen; Lee, Sui Mae

    2016-01-01

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

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

    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.

  11. Comparison of survival of Campylobacter jejuni in the phyllosphere with that in the rhizosphere of spinach and radish plants.

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

    2004-02-01

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

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

    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

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

    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...

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

    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

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

    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

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

    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

  17. Genotypic characterisation and cluster analysis of Campylobacter jejuni isolates from domestic pets, human clinical cases and retail food

    Acke Els; Carroll Cyril; O'Leary Aoife; McGill Kevina; Kelly Lorraine; Lawlor Amanda; Madden Robert H; Moran Lynn; Scates Pam; McNamara Eleanor; Moore John E; Jones Boyd R; Fanning Seamus; Whyte Paul

    2011-01-01

    Abstract The genetic similarity of Campylobacter jejuni isolates from pets, compared to human clinical cases and retail food isolates collected in Ireland over 2001-2006 was investigated by cluster analysis of pulsed-field gel electrophoresis (PFGE) fingerprinting profiles. Comparison of the PFGE profiles of 60 pet isolates and 109 human isolates revealed that seven (4.1%) profiles were grouped in clusters including at least one human and one pet C. jejuni isolate. In total six (1.6%) of 60 p...

  18. Structure of an unusual S-adenosylmethionine synthetase from Campylobacter jejuni.

    Zano, Stephen P; Pavlovsky, Alexander G; Viola, Ronald E

    2014-02-01

    S-Adenosylmethionine (AdoMet) participates in a wide range of methylation and other group-transfer reactions and also serves as the precursor for two groups of quorum-sensing molecules that function as regulators of the production of virulence factors in Gram-negative bacteria. The synthesis of AdoMet is catalyzed by AdoMet synthetases (MATs), a ubiquitous family of enzymes found in species ranging from microorganisms to mammals. The AdoMet synthetase from the bacterium Campylobacter jejuni (cjMAT) is an outlier among this homologous enzyme family, with lower sequence identity, numerous insertions and substitutions, and higher catalytic activity compared with other bacterial MATs. Alterations in the structure of this enzyme provide an explanation for its unusual dimeric quaternary structure relative to the other MATs. Taken together with several active-site substitutions, this new structure provides insights into its improved kinetic properties with alternative substrates. PMID:24531478

  19. 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.

    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...

  20. In Silico Analysis of the cadF Gene and Development of a Duplex Polymerase Chain Reaction for Species-Specific Identification of Campylobacter jejuni and Campylobacter coli

    Shams, Saeed; Bakhshi, Bita; Tohidi Moghadam, Tahereh

    2016-01-01

    Background Campylobacteriosis is a zoonotic infectious disease caused by Campylobacter jejuni and C. coli. The cadF gene is considered as a genus-specific gene while other genes are mainly used for discrimination at the species level. Objectives This study aimed to analyze the cadF gene and to develop a duplex PCR assay for simultaneous detection of C. coli and C. jejuni, the two commonly encountered species. Materials and Methods In silico analysis of the cadF gene was carried out by several software and available online tools. A duplex PCR optimized with specific primers was used for detection and differentiation of both species. To evaluate specificity and sensitivity of the test, a panel of different Campylobacter spp. together with several intestinal bacterial pathogens was tested. The limit of detection (LOD) of method was determined using serial dilutions of standard genomes. Results The analysis of the full size cadF gene indicated variations in this gene, which can be used to differentiate C. jejuni and C. coli. The duplex PCR designed in this study showed that it could simultaneously detect and differentiate both C. jejuni and C. coli with product sizes of 737 bp and 461 bp, respectively. This assay, with 100% specificity and sensitivity, had a limit of detection (LOD) of about 14 and 0.7 µg/mL for C. jejuni and C. coli, respectively. Conclusions In silico analysis of the cadF full-gene showed variations between the two species that can be used as a molecular target for differentiating C. jejuni and C. coli in a single-step duplex-PCR assay with high specificity and sensitivity. PMID:27127589

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

    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

  2. Serologic host response to Helicobacter pylori and Campylobacter jejuni in socially housed Rhesus macaques (Macaca mulatta

    Kienesberger Sabine

    2012-08-01

    Full Text Available Abstract Background Helicobacter pylori are successful colonizers of the human gastric mucosa. Colonization increases the risk of peptic ulcer disease and adenocarcinoma. However, potential benefits of H. pylori colonization include protection against early-onset asthma and against gastrointestinal infections. Campylobacter jejuni are a leading cause of bacterial diarrhea and complications include Guillain-Barré syndrome. Here, we describe the development of reliable serological assays to detect antibodies against those two bacteria in Rhesus macaques and investigated their distribution within a social group of monkeys. Methods Two cohorts of monkeys were analyzed. The first cohort consisted of 30 monkeys and was used to establish an enzyme-linked immunosorbent assay (ELISA for H. pylori antibodies detection. To evaluate colonization of those macaques, stomach biopsies were collected and analyzed for the presence of H. pylori by histology and culture. C. jejuni ELISAs were established using human serum with known C. jejuni antibody status. Next, plasma samples of the 89 macaques (Cohort 2 were assayed for antibodies and then statistically analyzed. Results An H. pylori IgG ELISA, which was 100% specific and 93% sensitive, was established. In contrast, the IgA ELISA was only 82% specific and 61% sensitive. The CagA IgG assay was 100% sensitive and 61% of the macaques were positive. In cohort 2, 62% macaques were H. pylori sero-positive and 52% were CagA positive. The prevalence of H. pylori IgG and CagA IgG increased with monkey age as described for humans. Of the 89 macaques 52% showed IgG against C. jejuni but in contrast to H. pylori, the sero-prevalence was not associated with increasing age. However, there was a drop in the IgG (but not in IgA mean values between infant and juvenile macaques, similar to trends described in humans. Conclusions Rhesus macaques have widespread exposure to H. pylori and C. jejuni, reflecting their social

  3. Distribution of flagella secreted protein and integral membrane protein among Campylobacter jejuni isolated from Thailand

    Pootong Piyarat

    2011-07-01

    Full Text Available Abstract Background Campylobacter jejuni, a gram-negative bacterium, is a frequent cause of gastrointestinal food-borne illness in humans throughout the world. There are several reports that the virulence of C. jejuni might be modulated by non-flagellar proteins that are secreted through the filament. Recently, FspA (Flagella secreted proteins have been described. Two alleles of fspA (fspA1 and fspA2 based on sequence analysis were previously reported and only the fspA2 allele was found in Thai isolates. The aim of this study is to analyze the deduced amino acid sequences fspA and the adjacent putative integral membrane protein from 103 Thai C. jejuni isolates. Results A total of 103 representative C. jejuni isolates were amplified by PCR for the fspA gene and the adjacent integral membrane protein gene. Two PCR product sizes were amplified using the same primers, an approximately 1600-bp PCR product from 19 strains that contained fspA and integral membrane protein genes and an approximately 800-bp PCR product from 84 strains that contained only the fspA gene. DNA sequencing was performed on the amplified products. The deduced amino acid sequences of both genes were analyzed separately using CLC Free Workbench 4 software. The analysis revealed three groups of FspA. Only FspA group 1 sequences (19/103 (corresponding to fspA1 consisting of 5 subgroups were associated with the adjacent gene encoding the integral membrane protein. FspA group 2 was the largest group (67/103 consisting of 9 subgroups. FspA group 2p (17/103 consisting of 7 subgroups was found to contain stop codons at a position before the terminal 142 position. Conclusions This study reveals greater heterogeneity of FspA (group 1, 2 and 2p among Thai C. jejuni isolates than previously reported. Furthermore, the subgroups of FspA groups 1 were associated with groups of integral membrane protein. The significance of these different FspA variants to virulence requires further study.

  4. Major Role for FeoB in Campylobacter jejuni Ferrous Iron Acquisition, Gut Colonization, and Intracellular Survival

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

    2006-01-01

    To assess the importance of ferrous iron acquisition in Campylobacter physiology and pathogenesis, we disrupted and characterized the Fe2+ iron transporter, FeoB, in Campylobacter jejuni NCTC 11168, 81-176, and ATCC 43431. The feoB mutant was significantly affected in its ability to transport 55Fe2+. It accumulated half the amount of iron than the wild-type strain during growth in an iron-containing medium. The intracellular iron of the feoB mutant was localized in the periplasmic space versu...

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

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

    2012-01-01

    Campylobacter spp. is the most common cause of bacterial diarrhoea in humans worldwide. Therefore, rapid and reliable methods fordetection and quantification of this pathogen are required. In this study, we have developed a reverse transcription quantitative real-time PCR(RT-qPCR) for detection and...... 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....

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

    N. CORCIONIVOSCHI

    2013-01-01

    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,...

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

    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.

  8. Probiotics down-regulate flaA sigma28 promoter in Campylobacter jejuni.

    Ding, Wu; Wang, Haifeng; Griffiths, Mansel W

    2005-11-01

    Lactobacilli and bifidobacteria are important members of the gastrointestinal microflora of humans and animals and are thought to have positive effects on human health. Therefore, there is an increasing interest in using these microorganisms as probiotics to be incorporated into either fermented dairy products or tablets. However, convincing scientific data that support claims of their health benefits are scarce. The effect of cell-free extracts of milk fermented by 10 probiotic bacteria (five Bifidobacterium strains and five Lactobacillus strains) on the expression of the flaA gene of Campylobacter jejuni was assessed using a fusion between the flaA sigma28 promoter and a promoterless luxCDABE cassette carried on the plasmid pRYluxCDABE, which resulted in strains with quantifiable luminescence linked to flaA sigma28 promoter activity. Cell-free extracts of milk fermented by all of the tested probiotic strains inhibited the growth of the C. jejuni and down-regulatedflaA sigma28 promoter activity. Two nonprobiotic lactic acid bacterial strains, Lactococcus lactis and Streptococcus thermophilus, were less inhibitory. PMID:16300065

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

    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.

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

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

    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 P21212, 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

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

    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

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

    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.

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

    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.

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

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

    Birk, Tina; Rosenquist, Hanne; Brondsted, L.; Ingmer, H.; Bysted, Anette; Christensen, Bjarke Bak

    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......, cells were suspended in chicken juice or brain heart infusion broth (BHIB) and added to 4-cm(2) skin pieces, which were subsequently stored at -18 degrees C. In the liquid model, cells were suspended in chicken juice or BHIB and stored at -18 degrees C. The decrease in the number of viable C. jejuni...... commercially processed chickens. The survival at -18 degrees C in the liquid model was also tested for three poultry isolates and three human clinical isolates of the serotypes 1.44, 2, and 4 complex. As observed for C. jejuni NCTC 11168, all the strains survived significantly better in chicken juice than in...

  17. Genotypic characterisation and cluster analysis of Campylobacter jejuni isolates from domestic pets, human clinical cases and retail food

    Acke Els

    2011-03-01

    Full Text Available Abstract The genetic similarity of Campylobacter jejuni isolates from pets, compared to human clinical cases and retail food isolates collected in Ireland over 2001-2006 was investigated by cluster analysis of pulsed-field gel electrophoresis (PFGE fingerprinting profiles. Comparison of the PFGE profiles of 60 pet isolates and 109 human isolates revealed that seven (4.1% profiles were grouped in clusters including at least one human and one pet C. jejuni isolate. In total six (1.6% of 60 pet and 310 food profiles were in clusters with at least one food and one pet C. jejuni isolate. The detection of only a small number of genetically indistinguishable isolates by PFGE profile cluster analysis from pets and from humans with enteritis in this study suggests that pets are unlikely to be an important reservoir for human campylobacteriosis in Ireland. However, genetically indistinguishable isolates were detected and C. jejuni from pets may circulate and may contribute to clinical infections in humans. In addition, contaminated food fed to pets may be a potential source of Campylobacter infection in pets, which may subsequently pose a risk to humans.

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

    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.

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

    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.

  20. Distribution of Campylobacter jejuni Penner serotypes in broiler flocks 1998-2000 in a small Danish community with special reference to serotype 4-complex

    Wedderkopp, A.; Nielsen, E.M.; Pedersen, Karl

    2003-01-01

    During the period January 1998-December 2001, all Danish broiler flocks were monitored bacteriologically for thermophilic campylobacters and isolates were stored at -80 degreesC. Six neighbouring broiler farms in a small community were selected for detailed examination of all Campylobacter jejuni...... isolated (n = 180) from these farms during 1998-2000 using Penner serotyping and pulsed-field gel electrophoresis (PFGE). The area and the farms were selected according to their prevalence of campylobacter so that both farms with low and high frequencies of campylobacter positive flocks were included in...... the study. The frequency of campylobacter positive flocks on the six farms ranged from 24.5 to 72.7%. One hundred and eighty of the isolates were C. jejuni (included in this study), 14 isolates were C. coli whereas 7 isolates belonged to other species but were not further identified. By serotyping of...

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

    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

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

    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.

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

    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.

  4. Role of flagella in adherence, internalization, and translocation of Campylobacter jejuni in nonpolarized and polarized epithelial cell cultures.

    Grant, C C; Konkel, M E; Cieplak, W; Tompkins, L S

    1993-01-01

    Previous studies of Campylobacter jejuni have suggested that flagellin is an adhesin for epithelial cells and that motility is a virulence factor of this bacterium. The role of flagella in the interactions of C. jejuni with nonpolarized and polarized epithelial cells was examined with flagellar mutants. Flagellated, nonmotile (flaA flaB+ Mot-) and nonflagellated, nonmotile (flaA flaB Mot-) mutants of C. jejuni were constructed by in vivo homologous recombination and gene replacement techniques. Both classes of mutants were found to adhere to cells of human epithelial origin (INT 407) equally well; however, on the basis of the percentage of the inoculum internalized, internalization of the flaA flaB Mot- mutants was decreased by factors ranging from approximately 30 to 40 compared with the parent. The flaA flaB+ Mot- mutant was internalized by the INT 407 cells at levels six- to sevenfold higher than the flaA flaB Mot- mutants. Both classes of mutants, unlike the parent, were unable to translocate across polarized Caco-2 monolayers. These results indicate that flagella are not involved in C. jejuni adherence to epithelial cells but that they do play a role in internalization. Furthermore, the results suggest that either the motility of C. jejuni or the product of flaA is essential for the bacterium to cross polarized epithelial cell monolayers. Images PMID:8478066

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

    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.

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

    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

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

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

    2012-03-01

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

  8. Structural insights from random mutagenesis of Campylobacter jejuni oligosaccharyltransferase PglB

    Ihssen Julian

    2012-09-01

    Full Text Available Abstract Background Protein glycosylation is of fundamental importance in many biological systems. The discovery of N-glycosylation in bacteria and the functional expression of the N-oligosaccharyltransferase PglB of Campylobacter jejuni in Escherichia coli enabled the production of engineered glycoproteins and the study of the underlying molecular mechanisms. A particularly promising application for protein glycosylation in recombinant bacteria is the production of potent conjugate vaccines where polysaccharide antigens of pathogenic bacteria are covalently bound to immunogenic carrier proteins. Results In this study capsular polysaccharides of the clinically relevant pathogen Staphylococcus aureus serotype 5 (CP5 were expressed in Escherichia coli and linked in vivo to a detoxified version of Pseudomonas aeruginosa exotoxin (EPA. We investigated which amino acids of the periplasmic domain of PglB are crucial for the glycosylation reaction using a newly established 96-well screening system enabling the relative quantification of glycoproteins by enzyme-linked immunosorbent assay. A random mutant library was generated by error-prone PCR and screened for inactivating amino acid substitutions. In addition to 15 inactive variants with amino acid changes within the previously known, strictly conserved WWDYG motif of N-oligosaccharyltransferases, 8 inactivating mutations mapped to a flexible loop in close vicinity of the amide nitrogen atom of the acceptor asparagine as revealed in the crystal structure of the homologous enzyme C. lari PglB. The importance of the conserved loop residue H479 for glycosylation was confirmed by site directed mutagenesis, while a change to alanine of the adjacent, non-conserved L480 had no effect. In addition, we investigated functional requirements in the so-called MIV motif of bacterial N-oligosaccharyltransferases. Amino acid residues I571 and V575, which had been postulated to interact with the acceptor peptide, were

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

    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

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

    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 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 luci...

  11. Demonstration of lipopolysaccharide with O-polysaccharide chains among different heat-stable serotypes of Campylobacter jejuni by silver staining of polyacrylamide gels.

    Blake, D C; Russell, R G

    1993-01-01

    Lipopolysaccharide (LPS) from three human and four monkey isolates of Campylobacter jejuni was extracted in high yields that revealed ladder-like structures in polyacrylamide gels by direct silver staining. These observations demonstrate that C. jejuni possesses LPS with O-chain repeating units typical of the family Enterobacteriaceae. The isolates showed differences in the number and electrophoretic mobility of bands in silver-stained gels.

  12. Cloning, sequencing, and expression of a gene from Campylobacter jejuni encoding a protein (Omp18) with similarity to peptidoglycan-associated lipoproteins.

    Konkel, M E; Mead, D J; Cieplak, W

    1996-01-01

    A Campylobacter jejuni genomic plasmid library was screened with antiserum generated against whole C. jejuni, revealing two immunoreactive clones. Sequence analysis of the recombinant plasmids revealed a common open reading frame of 498 nucleotides encoding a protein of 165 amino acids with a calculated molecular mass of 18,018 Da. The recombinant product partitioned to the outer membrane fractions of Escherichia coli transformants and has been designated Omp18. The deduced amino acid sequenc...

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

    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...

  14. Caracterização de isolados de Campylobacter jejuni de origem animal e humana quanto aos seus factores genéticos de virulência

    Santos, José Filipe Abreu dos, 1988

    2011-01-01

    Campylobacter jejuni é o principal agente causal de gastroenterite, sendo a carne de aves apontada como a principal fonte de infecção. O último relatório da EFSA indicou que a notificação de campilobacteriose aumentou em 2009 continuando a ser a zoonose mais comum na UE. O objectivo deste estudo foi avaliar a presença de diversos genes de virulência em isolados de Campylobacter jejuni, de origem animal (frangos, n = 37), de diferentes sistemas de produção (intensivo, extensivo e biológico), b...

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

    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.

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

    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.

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

    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.

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

    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.

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

    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

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

    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.

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

    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.

  2. Cloning, sequencing, and expression of a gene from Campylobacter jejuni encoding a protein (Omp18) with similarity to peptidoglycan-associated lipoproteins.

    Konkel, M E; Mead, D J; Cieplak, W

    1996-01-01

    A Campylobacter jejuni genomic plasmid library was screened with antiserum generated against whole C. jejuni, revealing two immunoreactive clones. Sequence analysis of the recombinant plasmids revealed a common open reading frame of 498 nucleotides encoding a protein of 165 amino acids with a calculated molecular mass of 18,018 Da. The recombinant product partitioned to the outer membrane fractions of Escherichia coli transformants and has been designated Omp18. The deduced amino acid sequence of the cloned C. jejuni gene exhibits considerable similarity to peptidoglycan-associated lipoproteins from other gram-negative bacteria. PMID:8613402

  3. Numerical analysis of DNA microarray data of Campylobacter jejuni strains correlated with survival, cytolethal distending toxin and haemolysin analyses

    On, Stephen L.W.; Dorrell, N.; Petersen, L.;

    2006-01-01

    Molecular epidemiological studies of the enteric pathogen Campylobacter jejuni have suggested that not all animal isolates are equally pathogenic to humans. We examined the use of numerical analysis of whole-genomotype data as a potential tool for evaluating C. jejuni virulence potential. Whole......-genome microarray analysis was used to determine the gene-level complementarity of 12 Danish strains to the pathogenic, genome-sequenced strain NCTC 11168. Cytolethal distending toxin (CDT) and haemolysin activities, and survival characteristics under aerobic conditions at room temperature were also determined....... Among the strains examined, 439 genes were polymorphic. Numerical analysis of these data by use of the squared Euclidean distance coefficient and Ward's clustering method clearly delineated strains into two clusters. CDT and haemolysin activities of cluster 1 strains were not statistically significantly...

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

    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

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

    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...... (measured as heterotrophic plate counts, HPC) were investigated in microcosms at 7 °C, containing non-sterilised drinking water, drinking water sediment and A. aquaticus collected from a non-chlorinated ground water based drinking water supply system. Concentrations of E. coli, K. pneumoniae and C. jejuni......Water lice, Asellus aquaticus (isopoda), frequently occur in drinking water distribution systems where they are a nuisance to consumers and water utilities. Whether they are solely an aesthetic problem or also affect the microbial water quality is a matter of interest. We studied the influence of A...

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

    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

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

    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.

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

    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

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

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

    2016-01-01

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

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

    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

  11. The use of FTA cards for transport and detection of gyrA mutation of Campylobacter jejuni from poultry.

    Sierra-Arguello, Y M; Faulkner, O; Tellez, G; Hargis, B M; Pinheiro do Nascimento, V

    2016-04-01

    The purpose of the present study was to evaluate a technique involving the use of commercially available FTA classic card (Whatman) for transporting and detection of DNA to use in PCR analysis and genetic sequencing of Campylobacter jejuni of poultry origin. Fifty isolates of Campylobacter jejuni were obtained from broiler carcasses in Rio Grande do Sul, Brazil. Antimicrobial susceptibility testing to ciprofloxacin revealed that all 50 isolates were resistant to ciprofloxacin. Each isolate was transferred to Brucella broth tubes and incubated overnight at 41.5°C. Cell cultures were diluted to match a McFarland Turbidity Standard 0.5, and 110 μL of the cell suspension were applied to one circle on Whatman FTA classic cards. The samples were then covered and allowed to dry at room temperature. Cards were identified and stored at room temperature until further use (3 mo after collection). FTA cards were shipped for analysis to the Department of Poultry Science, University of Arkansas. Amplification of the Campylobacter gyrA gene was successful and demonstrated strong bands for a large amplicon for all 50 samples preserved on FTA cards. Mutations present in each gene were confirmed by DNA sequencing. Then, 7 samples were chosen for the sequencing. The detection of a mutation regarding ciprofloxacin-resistant isolates revealed that 7 samples had a mutation in the gyrA gene. In conclusion, the characteristics of the profiles suggest that the DNA has maintained its integrity after 3 mo of storage at room temperature and is a suitable template for PCR and sequencing from Campylobacter samples. The application of this technology has potential in numerous methodologies, especially when working in remote areas and in developing countries where access to laboratory facilities and equipment is limited. PMID:26769268

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

    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.

  13. Use of Comparative Genomics and Eukaryotic Cell Adherence/Invasion Assays of the Food-Borne Pathogen Campylobacter jejuni for the Identification of Putative Virulence Factors

    Campylobacter, the leading bacterial etiology of acute humans gastroenteritis, is commonly associated with the handling/consumption of poultry. Eukaryotic cell adhesion/invasion assays were performed on fifty-one C. jejuni isolates. A range of adhesion/invasion abilities was exhibited. To identify ...

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

    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.

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

    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.

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

    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.

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

    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.

  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

    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. OCORRÊNCIA DE Campylobacter jejuni EM LEITE CRU E PASTEURIZADO COMERCIALIZADO EM CURITIBA, ESTADO DO PARANÁ, BRASIL

    Danielle Karen Narcizo

    2014-10-01

    Full Text Available Campylobacter jejuni é uma das bactérias mais envolvidas em surtos de origem alimentar em todo o mundo. A campilobacteriose é comumente associada ao consumo de carne de aves contaminadas, porém, existem relatos de vários surtos desta doença associados à ingestão de leite contaminado, em geral por falhas no processo de pasteurização, ou mesmo pelo consumo de leite cru. Objetivou-se avaliar a ocorrência de C. jejuni em amostras de leite cru ou pasteurizado, bem como avaliar diferentes metodologias de análise para detecção deste microrganismo. Foram analisadas 30 amostras de leite, sendo 15 amostras de leite cru e 15 amostras de leite pasteurizado, as quais foram adquiridas na região metropolitana de Curitiba-PR, entre setembro de 2012 e março de 2013. As amostras foram submetidas à contagem de C. jejuni por meio do método de plaqueamento direto e pesquisa após o enriquecimento seletivo. Em nenhuma das amostras de leite pasteurizado foi detectada a presença de C. jejuni por nenhum método utilizado. Por meio do método de plaqueamento direto, foram identificadas duas amostras de leite cru contaminadas por C. jejuni, com contagens entre 4,0 x 101 a 1,65 x 102 UFC/mL. Pelo método de enriquecimento seletivo, C. jejuni foi detectado em quatro amostras de leite cru, indicando a maior eficiência do método qualitativo em função da recuperação de células injuriadas e, portanto, maior confiabilidade para detecção deste patógeno. A ocorrência de leite contaminado por C. jejuni constitui um alerta para orientação de medidas preventivas no controle e eliminação deste patógeno na cadeia de produção e beneficiamento do produto.

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

    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 ...

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

    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.

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

    Ranjitkar, Samir; Engberg, Ricarda Margarete

    2016-04-01

    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. PMID:27100153

  3. Polar flagellar biosynthesis and a regulator of flagellar number influence spatial parameters of cell division in Campylobacter jejuni.

    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

  4. 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

    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.

  5. Eugenol wash and chitosan based coating reduces Campylobacter jejuni counts on poultry products

    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 ...

  6. Gene Expression Profile of Campylobacter jejuni in Response to Chicken Weep

    Campylobacter spp. are one of the most common causes of foodborne bacterial gastroenteritis. Handling and consumption of raw poultry products are considered to be a major source of Campylobacter induced disease in humans. There is a high incidence of Campylobacter-positive poultry carcasses, even ...

  7. Differential carbon source utilization by Campylobacter jejuni strain 11168 in response to growth temperature variation

    Campylobacter spp. readily colonize the intestinal tracts of both human and avian species. While most often a commensal organism in birds, campylobacters remain the leading cause of bacterial gastroenteritis in humans. The association of campylobacters with poultry is well established as a primary...

  8. Genetic diversity among Campylobacter jejuni isolates from healthy livestock and their links to human isolates in Spain.

    Oporto, B; Juste, R A; López-Portolés, J A; Hurtado, A

    2011-08-01

    This study was aimed at determining the genetic diversity of Campylobacter jejuni from healthy ruminants and poultry, and study by multilocus sequence typing (MLST) their links to human isolates in Spain. MLST analysis of 160 animal isolates generated 45 sequence types (STs, nine of them new to this study), that clustered into 18 clonal complexes (CC) and nine singletons. The 71 isolates from humans generated 28 STs (13 CC plus four singletons). Only 11 STs and nine CCs were shared by humans and animals (particularly from dairy cattle and sheep), mainly corresponding to sporadic cases rather than outbreaks, probably as an adaptation of the general human population to the types commonly circulating in livestock. PCR analysis of the distribution of four virulence-associated genes detected the cdtABC gene cluster in all 160 isolates but with a 700-bp deletion in four of them, and amplified the virB11, cgtB and wlaN genes in 4.7%, 21.3% and 21.9% respectively. A subset of 87 C. jejuni animal isolates analysed using flaA PCR-RFLP, MLST and pulsed-field gel electrophoresis generated 31, 38 and 55 types respectively. The combined typing approach used provided reliable inter-strain relationships, confirming the co-existence of several strains in some farms, but also identifying identical genotypes sampled over a wide temporal span in different environments and hosts. Typing results confirmed a high genetic diversity of C. jejuni in our region and suggested that ruminants are also important sources for human infection. MLST data provided will help to obtain a more comprehensive image of the population structure of C. jejuni and establish reliable source attribution schemes. PMID:21040505

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

    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...

  10. The Bacteriophage Carrier State of Campylobacter jejuni Features Changes in Host Non-coding RNAs and the Acquisition of New Host-derived CRISPR Spacer Sequences

    Hooton, Steven P. T.; Brathwaite, Kelly J.; Connerton, Ian F.

    2016-01-01

    Incorporation of self-derived CRISPR DNA protospacers in Campylobacter jejuni PT14 occurs in the presence of bacteriophages encoding a CRISPR-like Cas4 protein. This phenomenon was evident in carrier state infections where both bacteriophages and host are maintained for seemingly indefinite periods as stable populations following serial passage. Carrier state cultures of C. jejuni PT14 have greater aerotolerance in nutrient limited conditions, and may have arisen as an evolutionary response to selective pressures imposed during periods in the extra-intestinal environment. A consequence of this is that bacteriophage and host remain associated and able to survive transition periods where the chances of replicative success are greatly diminished. The majority of the bacteriophage population do not commit to lytic infection, and conversely the bacterial population tolerates low-level bacteriophage replication. We recently examined the effects of Campylobacter bacteriophage/C. jejuni PT14 CRISPR spacer acquisition using deep sequencing strategies of DNA and RNA-Seq to analyze carrier state cultures. This approach identified de novo spacer acquisition in C. jejuni PT14 associated with Class III Campylobacter phages CP8/CP30A but spacer acquisition was oriented toward the capture of host DNA. In the absence of bacteriophage predation the CRISPR spacers in uninfected C. jejuni PT14 cultures remain unchanged. A distinct preference was observed for incorporation of self-derived protospacers into the third spacer position of the C. jejuni PT14 CRISPR array, with the first and second spacers remaining fixed. RNA-Seq also revealed the variation in the synthesis of non-coding RNAs with the potential to bind bacteriophage genes and/or transcript sequences. PMID:27047470

  11. Structural Context for Protein N-glycosylation in Bacteria: The Structure of PEB3, an Adhesin from Campylobacter Jejuni

    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.

  12. Research progress on flagella of Campylobacter jejuni%空肠弯曲菌鞭毛研究进展

    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.

  13. Efficacy of flagellin gene typing for epidemiological studies of Campylobacter jejuni in poultry estimated by comparison with macrorestriction profiling

    Petersen, L.; On, S.L.W.

    2000-01-01

    isolates were obtained. The PCR-Fla type of one isolate changed spontaneously after five subcultures, illustrating the relative plasticity of the gene locus. Comparison of MRPs within and between Fla-types support the view that some PCR-Fla types may be conserved within clonal lines. It is concluded that......Thirty isolates of Campylobacter jejuni isolated from 29 different Danish broiler flocks were chosen for the evaluation of PCR-Fla typing as a genotyping tool. Except for two isolates that originated from the same broiler flock, the isolates were clearly distinguishable on basis of their...... macrorestriction profiles using the restriction endonucleases SmaI and KpnI. PCR-Fla typing of the 30 isolates yielded 16 distinct genotypes, whereas one isolate was untypeable by this method. The dominant PCR-Fla type (1/1) was shared by eight isolates, and five additional Fla groups containing two or three...

  14. 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

    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.

  15. Analysis of putative chemoreceptor proteins of Campylobacter jejuni

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

    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...... putative chemoreceptors (tlp1, tlp2, tlp3, docB and docC) in the sequenced strain C. jejuni NCTC11168. The chemotactic capacity of these mutants were investigated by quantifying bacterial accumulation in capillaries filled with attractants or repellants and by measuring migration zones towards or away from...

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

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

    2006-01-01

    was observed for chloramphenicol, nalidixic acid and ciprofloxacin (P <0.05). The trends in resistance in the period 1996-2003 among C jejuni isolates from chicken meat indicate a decrease in the occurrence of resistance towards fluoroquinolones. This may be due to reduced application of...... fluoroquinolones for food animals. Monitoring of the occurrence of antimicrobial resistance in C. jejuni isolated from raw uncooked poultry has been performed on a yearly basis since 1996, thus providing useful insight into consumer exposure to antimicrobial-resistant C. jejuni....

  17. Distribution of nine virulence-associated genes in Campylobacter jejuni and C. coli isolated from broiler feces in Shiraz, Southern Iran.

    Khoshbakht, Rahem; Tabatabaei, Mohammad; Hosseinzadeh, Saeid; Shekarforoush, Seied Shahram; Aski, Hesamaddin Shirzad

    2013-09-01

    To investigate the prevalence of nine virulence and toxin genes of Campylobacter, a total of 90 Campylobacter strains including 48 C. jejuni and 42 C. coli were recovered from chicken feces by cultivation methods. The isolates were identified on the basis of polymerase chain reaction (PCR) detection of 16SrRNA and multiplex PCR for determining two species. For confirmed strains, PCR was carried out for the presence of virulence genes using specific primers. Data were analyzed by SPSS software, version 12.0.1. The cadF gene and three genes associated with cytolethal distending toxin were present in 100% of isolates. Plasmid virB11 gene was not found in any of the Campylobacter isolates, and the prevalence of pldA, wlaN, iamA, and cgtB genes were 92.22%, 82.22%, 81.11%, and 22.22%, respectively. The findings revealed that the distribution of the majority of these genes were not dissimilar among Campylobacter species. The results emphasized that many of the pathogenic C. jejuni and C. coli may have these genes, and the Campylobacter strains with poultry origin have pathogenic potential properties for humans. PMID:23789768

  18. Antimicrobial, Rheological, and Thermal Properties of Plasticized Polylactide Films Incorporated with Essential Oils to Inhibit Staphylococcus aureus and Campylobacter jejuni.

    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

  19. Genetic diversity of Campylobacter jejuni isolates from Korea and travel-associated cases from east and southeast Asian countries.

    Cha, Injun; Kim, Nan-Ok; Nam, Jung-Gu; Choi, Eun-Suk; Chung, Gyung Tae; Kang, Yeon-Ho; Hong, Sahyun

    2014-01-01

    Forty domestic and travel-associated Campylobacter jejuni isolates were analyzed by profiling 7 pathogenic genes (cdtB, cadF, Cj0131, ciaB, racR, wlaN, and virB11) along with multilocus sequence typing (MLST) and antimicrobial susceptibility testing. cdtB, cadF, and Cj0131 were present in all isolates, whereas virB11 was not detected in either domestic or travel-associated isolates. ciaB was present in all domestic isolates and 94% of travel-associated isolates. The respective detection rates of racR and wlaN in domestic and travel-associated isolates were 94% and 71% and 35.3% and 23%, respectively. MLST analyses of the 40 isolates generated 25 different sequence types (STs). ST-443 (12 isolates) and ST-21 (8 isolates) were dominant among the domestic isolates; however, STs varied among travel-associated isolates. Nalidixic acid, tetracycline, and ciprofloxacin resistance rates of the 40 isolates were 100% (40/40), 95% (38/40), and 88% (35/40), respectively. Domestic isolates exhibited 2-fold higher ciprofloxacin, telithromycin, and chloramphenicol resistance rates than travel-associated isolates. These results indicate a diverse genetic background for travel-associated C. jejuni and suggest that this pathogen may be an important emerging public health threat to travelers. PMID:25410568

  20. Phenotypic and Genotypic Evidence for L-fucose Utilization by Campylobacter jejuni

    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....

  1. Transducer Like Proteins of Campylobacter jejuni 81-176: Role in chemotaxis and colonization of the chicken gastrointestinal tract

    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.

  2. The impact of the LuxS mutation on phenotypic expression of factors critical for Campylobacter jejuni colonization.

    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

  3. AI-2 does not function as a quorum sensing molecule in Campylobacter jejuni during exponential growth in vitro

    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.

  4. Effect of refrigerated and frozen storage on the survival of Campylobacter jejuni in cooked chicken meat breast.

    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

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

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

    2016-01-01

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

  6. Attachment of Campylobacter jejuni on biofilms from two chicken houses in Thailand

    Daungjinda, M.

    2007-01-01

    Full Text Available The attachment of C. jejuni on four gram negative biofilms (FBRL-C04, FBRL-B05, FBRL-F01 and FBRL-B06 isolated from two chicken houses were studied. It was found that C. jejuni attached to biofilm of FBRL-F01 at the highest rate (4.4 logCFU/cm2 compared (P<0.05 to FBRL-C04 FBRL-B05 and FBRL-B06 (4.0 4.0 and 4.1 logCFU/cm2, respectively. Coaggregation between C. jejuni and biofilm organisms may indicate the ability of organisms to form biofilm together. Percent coaggregation between C. jejuni and biofilm organisms, FBRL-C04 and FBRL-F01 was 39.14% and 33.70%, respectively, higher (P<0.05 than thatwith FBRL-B05 and FBRL-B06 (-3.38% and 12.87%, respectively. Hydrophobicity of planktonic and biofilm cells of C. jejuni and 4 biofilm producers were measured by the microbial adhesion to hydrocarbon (MATH method using hexadecane. FBRL-B06 showed the highest (P<0.05 hydrophobicity (68.95% indicating more hydrophobic components on its cell surface. Planktonic cells had lower (P<0.05 hydrophobicity than biofilm cells. However, the degree of hydrophobicity of biofilm cells was not related to attachment of C. jejuni on biofilms.

  7. A fluoroquinolone resistance associated mutation in gyrA affects DNA supercoiling in Campylobacter jejuni

    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.

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

    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.

  9. De Novo Asymmetric Synthesis of a 6-O-Methyl-d-glycero-l-gluco-heptopyranose-Derived Thioglycoside for the Preparation of Campylobacter jejuni NCTC11168 Capsular Polysaccharide Fragments.

    Ashmus, Roger A; Jayasuriya, Anushka B; Lim, Ying-Jie; O'Doherty, George A; Lowary, Todd L

    2016-04-01

    An enantioselective de novo synthesis of a thioglycoside derivative of the 6-O-methyl-d-glycero-l-gluco-heptopyranose residue found in the Campylobacter jejuni NCTC11168 (HS:2) capsular polysaccharide is reported. The compound is obtained from a furfural-derived chiral diol in 11 steps. Notably, compared to the only previous synthesis of this molecule, this approach significantly reduces the number of purification steps required to obtain the target. PMID:26982173

  10. Survival of lactic acid and chlorine dioxide treated Campylobacter jejuni under suboptimal conditions of pH, temperature and modified atmosphere

    Smigic, Nada; Rajkovic, Andreja; Nielsen, Dennis Sandris;

    2010-01-01

    Campylobacter jejuni cells treated with lactic acid (LA, 3% lactic acid, pH 4.0, 2 min) or chlorine dioxide (ClO(2), 20 ppm, 2 min) were inoculated in Bolton broth (pH 6.0) and incubated under 80% O(2)/20% N(2), 80% CO(2)/20% N(2), air or micro-aerophilic (10% CO(2)/85% N(2)/5% O(2)) atmosphere, at 4 degrees C...

  11. Phase variation of a Type IIG restriction-modification enzyme alters site-specific methylation patterns and gene expression in Campylobacter jejuni strain NCTC11168

    Anjum, Awais; Kelly, Brathwaite; Aidley, Jack B; Connerton, Phillippa L.; Cummings, Nicola J; Parkhill, Julian; Ian F Connerton; Bayliss, Christopher D

    2016-01-01

    Phase-variable restriction-modification systems are a feature of a diverse range of bacterial species. Stochastic, reversible switches in expression of the methyltransferase produces variation in methylation of specific sequences. Phase-variable methylation by both Type I and Type III methyltransferases is associated with altered gene expression and phenotypic variation. One phase-variable gene of Campylobacter jejuni encodes a homologue of an unusual Type IIG restriction-modification system ...

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

    Skovgård, Henrik; Hald, Birthe

    % Campylobacter positive at 6, 12, 18 hours, respectively, which was significantly lower than the higher dose groups (ChiSq=40.20, P<0.0001). We conclude that the duration of the carriage period depended on temperature and inoculation dose, and was less than a day at 20 ºC and higher........6 x 107 C. jejuni cells directly on their mouthparts. Subsequently, the flies were incubated individually at 15, 20, 25, 30 and 35 ºC and tested for Campylobacter after 0, 6, 12, 18 or 24 hours. The test was done by incubation of each fly individually in Bolton broth (Oxoid) at 37 ºC. After 48 hours......, 10 µL brooth was streaked on modified cefoperazone charcoal deoxycholate agar plates (mCCDA) (Oxoid) which were incubated microaerobically at 42 ºC for 24 hours. For a dose response experiment at 25 ºC, house flies (n = 225) were inoculated in three series: 6.5 x 106, 6.0 x 104 and 8.2 x 102 C...

  13. Signal balancing by the CetABC and CetZ chemoreceptors controls energy taxis in Campylobacter jejuni.

    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.

  14. Calcium and protein kinase C play an important role in Campylobacter jejuni-induced changes in Na+ and Cl- transport in rat ileum in vitro.

    Kanwar, R K; Ganguly, N K; Kumar, L; Rakesh, J; Panigrahi, D; Walia, B N

    1995-04-24

    The pathophysiological mechanism of Campylobacter jejuni (enterotoxigenic) induced secretory diarrhoea remains least understood. To investigate the mechanism(s) involved, the unidirectional fluxes of Na+ and Cl- were measured across the C. jejuni live culture infected and control (non infected) rat ileum (unstriped), in vitro by Ussing technique under short circuit conditions, in the presence or absence of: Ca2+ ionophore A23187 (5 microM), 1-verapamil (100 microM), calmodulin (CaM) antagonist W-7 (100 microM), dantrolene (25 microM), protein kinase C (PKC) activator PMA (100 ng/ml) and H-7 (60 microM), selective inhibitor of PKC. There was net absorption of Na+ and enhanced Cl- secretion in infected animals while in control animals there was net absorption of Na+ and marginal secretion Cl-.Ca2+ ionophore A23187 mimicked the effects of C. jejuni infection whereas 1-verapamil had significant antisecretory effect on Na+ and Cl- secretion in infected animals. In vitro measurement of undirectional 45Ca fluxes in Ussing chamber experiments revealed net absorption of Ca2+ in infected rat ileum as compared to net secretion of Ca2+ in control rat ileum. These observations clearly indicate that there is increased stimulation of Ca2+ uptake from extracellular milieu to the enterocytes during C. jejuni-induced diarrhoea. The intracellular calcium levels (Ca2+]i (as measured by fluorescent probe Fura-2AM) were found to be raised significantly (P < 0.0001) in enterocytes isolated from C. jejuni infected ileum as compared to the enterocytes from control ileum. The observed increase in [Ca2+]i in enterocytes isolated from C. jejuni live culture supernatant treated rat ileum further shows the involvement of enterotoxin in diarrhoeal process. Dantrolene decreased significantly C. jejuni-induced net Na+ and Cl- secretion but it could not reverse it to absorption suggesting the partial involvement of Ca2+ mobilised from intracellular stores in mediating secretion. W-7 failed to

  15. Serotypes, molecular and antimicrobial characteristics of Campylobacter jejuni isolated from chicken meats in Northeastern Thailand during December, 2007 to June, 2008

    Tetsuo Asai

    2011-10-01

    Full Text Available Campylobacter species were isolated from chicken meats in northeastern Thailand from December, 2007 to June,2008. From 1,930 samples, Campylobacter spp. were obtained from 556 (28.8%. Campylobacter species were isolated fromchicken livers at higher percentages compared to the other parts of chicken meats (p<0.001. Among 294 Campylobacterisolates, 187 (63.6% were identified as C. jejuni, and 107 (36.4% as C. coli. The results of serotyping by Penner’s methodshowed that serotype L (22.7% and serotype A (18.7% were predominant. Antimicrobial susceptibility test revealed that90.7, 37.3, 29.3 and 13.3% of C. jejuni were resistant to OFLX, DOXY, EM and CP, respectively. MIC50/MIC90 of OFLX, DOXY,EM and CP were 16/128, 4/256, 0.5/128 and 2/64 μg/ml, respectively. Precaution needs to be emphasized when attempt to useOFLX and DOXY for veterinary and human medicine due to the high percentage of resistance among C. jejuni isolated fromchicken meat origin. These alarming figures should be notified to the general public.

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

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

    2010-07-01

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

  17. Intracellular pH as an indicator of viability and resuscitation of Campylobacter jejuni after decontamination with lactic acid

    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...... increased the subpopulation of the cells with positive pH gradient, mostly comprised of the cells with DeltapH>0.5, indicating the ability of C. jejuni cells to regulate their metabolic activity under suboptimal conditions.......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...... increased pH(i) and colony count (recovery study). On the contrary, 24 h incubation at suboptimal temperature of 4 degrees C, showed pH(i) decrease to pH(ex)=6.0 (no pH gradient) in the whole population of C. jejuni cells. Rather than dying, cells exposed for longer time (72 and 120 h) to 4 degrees C...

  18. 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

    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

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

    Birk, Tina; Grønlund, Anne Christine Jørgensen; Christensen, Bjarke Bak; Knochel, S.; Lohse, Kristin; Rosenquist, Hanne

    2010-01-01

    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, and...... soya sauce) revealed that such ingredients reduced counts of C. jejuni by at least 0.8 log units Oil meat medallions. Three low pH marinades (pH <3) based oil pomegranate syrup. lemon juice, and white wine vinegar were prepared. When applied in whole filets, these marinades resulted in a reduction of...

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

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

    2010-01-01

    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......, and soya sauce) revealed that such ingredients reduced counts of C. jejuni by at least 0.8 log units Oil meat medallions. Three low pH marinades (pH syrup. lemon juice, and white wine vinegar were prepared. When applied in whole filets, these marinades resulted in a reduction...

  1. 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

    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

  2. Synthesis of Antibodies-Conjugated Fluorescent Dye-Doped Silica Nanoparticles for a Rapid Single Step Detection of Campylobacter jejuni in Live Poultry

    The preparation of antibodies-conjugated fluorescent dye-doped silica nanoparticles (FDS-NPs) was developed to detect Campylobacter jejuni cells under a fluorescence microscope. The particles prepared by sol-gel microemulsion techniques have a round shape with an average size of 43±4 nm. They were highly photo stable and could emit strong orange fluorescent for 60 min. Both amine- and carboxyl-functionalized properties were evident from FTIR and FT Raman spectra. The FDS-NPs conjugated with antibodies against C. jejuni were well dispersed in PBS solution at 20 mM of NaCl. The conjugation with monoclonal antibodies against C. jejuni was successful. The direct observation of the antibodies-conjugated FDS-NPs- that bounds C. jejuni with Petroff Hausser counting chamber at 40 x was clear. The different focus lengths clearly separated bound and unbound FDS-NPs under the microscope. We successfully synthesis the bio-conjugated dye doped silica nanoparticles for C. jejuni that are easy to use and giving clear detection in due time.

  3. Synthesis of Antibodies-Conjugated Fluorescent Dye-Doped Silica Nanoparticles for a Rapid Single Step Detection of Campylobacter jejuni in Live Poultry

    Wachira Tansub

    2012-01-01

    Full Text Available The preparation of antibodies-conjugated fluorescent dye-doped silica nanoparticles (FDS-NPs was developed to detect Campylobacter jejuni cells under a fluorescence microscope. The particles prepared by sol-gel microemulsion techniques have a round shape with an average size of 43 ± 4 nm. They were highly photo stable and could emit strong orange fluorescent for 60 min. Both amine- and carboxyl-functionalized properties were evident from FTIR and FT Raman spectra. The FDS-NPs conjugated with antibodies against C. jejuni were well dispersed in PBS solution at 20 mM of NaCl. The conjugation with monoclonal antibodies against C. jejuni was successful. The direct observation of the antibodies-conjugated FDS-NPs- that bounds C. jejuni with Petroff Hausser counting chamber at 40x was clear. The different focus lengths clearly separated bound and unbound FDS-NPs under the microscope. We successfully synthesis the bio-conjugated dye doped silica nanoparticles for C. jejuni that are easy to use and giving clear detection in due time.

  4. 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

    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

  5. Campylobacter jejuni strains of human and chicken origin are invasive in chickens after oral challenge

    Knudsen, Katrine Nørrelund; Bang, Dang Duong; Andresen, Lars Ole; Madsen, Mogens

    2006-01-01

    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...

  6. Rapid paracellular transmigration of Campylobacter jejuni across polarized epithelial cells without affecting TER: role of proteolytic-active HtrA cleaving E-cadherin but not fibronectin

    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.

  7. Rapid paracellular transmigration of Campylobacter jejuni across polarized epithelial cells without affecting TER: role of proteolytic-active HtrA cleaving E-cadherin but not fibronectin

    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.

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

    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. PMID:27041390

  9. Structural analysis of PseH, the Campylobacter jejuni N-acetyltransferase involved in bacterial O-linked glycosylation

    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.

  10. Structural analysis of PseH, the Campylobacter jejuni N-acetyltransferase involved in bacterial O-linked glycosylation

    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

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

    Anna D Grabowska

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

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

    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...

  13. Analysis of the LIV System of Campylobacter jejuni Reveals Alternative Roles for LivJ and LivK in Commensalism beyond Branched-Chain Amino Acid Transport ▿

    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...

  14. Immunogenicety and immunoprotection of recombinant PEB1 in Campylobacter-jejuni-infected mice

    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.

  15. Detection of Campylobacter species using monoclonal antibodies

    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.

  16. Campylobacter jejuni adenosine triphosphate phosphoribosyltransferase is an active hexamer that is allosterically controlled by the twisting of a regulatory tail.

    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

  17. Guillain-Barré syndrome and Campylobacter jejuni infection%GBS与空肠弯曲菌感染

    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.

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

    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.

  19. Characterization of CetA and CetB, a bipartite energy taxis system in Campylobacter jejuni.

    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

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

    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.

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

    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.

  2. Whole-Genome Sequencing of a Campylobacter jejuni Strain Isolated from Retail Chicken Meat Reveals the Presence of a Megaplasmid with Mu-Like Prophage and Multidrug Resistance Genes

    Marasini, Daya

    2016-01-01

    Genome sequencing of Campylobacter jejuni strain T1-21 isolated from retail chicken meat revealed the presence of a chromosome of 1,565,978 bp and a megaplasmid of 82,732 bp that contains Mu-like prophage and multidrug resistance genes. This is the first reported sequence of a Campylobacter megaplasmid >55 kb. PMID:27231378

  3. Investigation and resistance analysis of Campylobacter jejuni in broiler%鸡源空肠弯曲菌调查与耐药性分析

    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

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

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

    2001-01-01

    , 314 C jejuni and 32 C coli isolates from parent and broiler flocks and from the surroundings of broiler houses were typed by flagellin gene PCR/RFLP fla-typing), and selected isolates were also typed by serotyping and macrorestriction profiling using PFGE (MRP/PFGE). The combined typing results showed...... were clearly discriminated by fla-typing as well as by MRP/PFGE, except for a few cases where individual isolates belonging to two different clones were found to have altered fla-types. Similarly, one C coli clone showed pronounced fla-type variation. The present results lead to the conclusion that...

  5. Bacteriophage F336 Recognizes the Capsular Phosphoramidate Modification of Campylobacter jejuni NCTC11168

    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......, we found that 11168R lacks an O-methyl phosphoramidate (MeOPN) moiety attached to the GalfNAc on the capsular polysaccharide (CPS), which was further confirmed by mass spectroscopy. Sequence analysis of 11168R showed that the potentially hypervariable gene cj1421, which encodes the GalfNAc Me...

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

    Wang, Zhenyu; Sekulovic, Andrea; Kutter, Jörg Peter; Bang, D.D.; Wolff, Anders

    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 Orange and TO-PRO-3, were added to the PCR mixture to realize the real-time PCR. The presented approach shows reliable real-time quantitative information of the PCR amplification of the targeted ge...

  7. Vaccines for viral and bacterial pathogens causing acute gastroenteritis: Part II: Vaccines for Shigella, Salmonella, enterotoxigenic E. coli (ETEC) enterohemorragic E. coli (EHEC) and Campylobacter jejuni.

    O'Ryan, Miguel; Vidal, Roberto; del Canto, Felipe; Carlos Salazar, Juan; Montero, David

    2015-01-01

    In Part II we discuss the following bacterial pathogens: Shigella, Salmonella (non-typhoidal), diarrheogenic E. coli (enterotoxigenic and enterohemorragic) and Campylobacter jejuni. In contrast to the enteric viruses and Vibrio cholerae discussed in Part I of this series, for the bacterial pathogens described here there is only one licensed vaccine, developed primarily for Vibrio cholerae and which provides moderate protection against enterotoxigenic E. coli (ETEC) (Dukoral(®)), as well as a few additional candidates in advanced stages of development for ETEC and one candidate for Shigella spp. Numerous vaccine candidates in earlier stages of development are discussed. PMID:25715096

  8. Rapid detection of Campylobacter jejuni by multiplex PCR technique%应用多重PCR快速检测空肠弯曲菌

    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

    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

    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 for their...... 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 of this...... 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. Impact of Campylobacter jejuni cj0268c knockout mutation on intestinal colonization, translocation, and induction of immunopathology in gnotobiotic IL-10 deficient mice.

    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

  12. Multiple factors interact to produce responses resembling spectrum of human disease in Campylobacter jejuni infected C57BL/6 IL-10-/- mice

    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. Inhibition of Campylobacter jejuni on fresh chicken breasts by κ-carrageenan/chitosan-based coatings containing allyl isothiocyanate or deodorized oriental mustard extract.

    Olaimat, Amin N; Fang, Yuan; Holley, Richard A

    2014-09-18

    Campylobacter species are common bacterial pathogens associated with human gastroenteritis worldwide. The objectives of this study were to determine the minimum inhibitory (MIC) and minimum bactericidal (MBC) concentrations of allyl isothiocyanate (AITC) against 4 Campylobacter jejuni strains in Mueller-Hinton (MH) broth at 4, 21, 37 and 42°C and to screen the C. jejuni strains for their ability to degrade sinigrin (which forms AITC) in pH7.0 MH broth at 35°C for 21d. Also evaluated was the antimicrobial activity of an edible 0.2% κ-carrageenan/2% chitosan-based coating containing AITC or deodorized oriental mustard extract against a 4 strain C. jejuni cocktail (6.2log10CFU/g) on vacuum-packaged fresh chicken breasts during 4°C storage. MIC values of AITC were 0.63 to 1.25ppm and 2.5 to 5ppm against tested strains at 37 and 42°C, respectively. However, the MBC was 2.5 and 5ppm at 37 and 42°C, respectively, and increased to a range of 40 to 160ppm at 4°C. κ-Carrageenan/chitosan-based coatings containing 50 or 100μl/g AITC reduced viable C. jejuni to undetectable levels on chicken breast after 5d at 4°C, while 25μl/g AITC or 200 to 300mg/g mustard extract in coatings reduced C. jejuni numbers by 1.75 to 2.78log10CFU/g more than control coatings without antimicrobial. Both oriental mustard extract (50 to 300mg/g) and AITC (≥25μl/g) reduced aerobic bacteria by 1.72 to 2.75log10CFU/g and lactic acid bacteria (LAB) by 0.94 to 3.36log10CFU/g by 21d compared to the control coating. κ-Carrageenan/chitosan coatings containing ≥50μl/g AITC or ≥300mg/g oriental mustard showed excellent potential to control C. jejuni viability on raw chicken. PMID:25058687

  14. Structural and Functional Analysis of Campylobacter jejuni PseG: a Udp-sugarhydrolase from the Pseudaminic Acid Biosynthetic Pathway

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

    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.

  16. A “successful allele” at Campylobacter jejuni contingency locus Cj0170 regulates motility; “successful alleles” at locus Cj0045 are strongly associated with mouse colonization

    Artymovich, Katherine; Kim, Joo-Sung; Linz, John E.; Hall, David F.; Kelley, Lauren E.; Kalbach, Harrison L.; Kathariou, Sophia; Gaymer, Jean; Paschke, Brenda

    2013-01-01

    Campylobacter jejuniis an important foodborne pathogen of humans and its primary reservoir is the gastrointestinal (GI) tract of chickens. Our previous studies demonstrated that phase variation to specific “successful alleles” at C. jejuni contingency loci Cj0045 (successful alleles carry 9G or 10G homopolymeric tracts) and Cj0170 (successful allele carries a 10G homopolymeric tract) in C. jejuni populations is strongly associated with colonization and enteritis in C57BL/6 IL-10 deficient mice. In the current study, we strengthened the association between locus Cj0170, Cj0045, and mouse colonization. We generated 8 independent strains derived from C. jejuni 11168 strain KanR4 that carried a Cj0170 gene disruption and these were all non motile. Two randomly chosen strains with the Cj0170 gene disruption (DM0170-2 and DM0170-6) were gavaged into mice. DM0170-2 and DM0170-6 failed to colonize mice while the control strain that carried a “successful” Cj0170 10G allele was motile and did colonize mice. In parallel studies, when we inoculated C. jejuni strain 33292 into mice, the “unsuccessful” Cj0045 11G allele experienced phase variation to “successful” 9G and 10G alleles in 2 independent experiments prior to d4 post inoculation in mice while the “successful” 9G allele in the control strain remained stable through d21 post inoculation or shifted to other successful alleles. These data confirm that locus Cj0170 regulates motility in C. jejuni strain KanR4 and is a virulence factor in the mouse model. The data also support a possible role of locus Cj0045 as a virulence factor in strain 33292 in infection of mice. PMID:23541212

  17. Comparison of lipooligosaccharide biosynthesis genes of Campylobacter jejuni strains with varying abilities to colonize the chicken gut and to invade Caco-2 cells.

    Müller, Jens; Meyer, Birgit; Hänel, Ingrid; Hotzel, Helmut

    2007-12-01

    Campylobacter jejuni strains develop a high variability of lipooligosaccharide (LOS) structures on the cell surface based on variations in the genetic content of the LOS biosynthesis locus. While the importance of these variations for ganglioside mimicry as a critical factor in the triggering of Guillain-Barré syndrome has already been shown, little work has been done on the investigation of LOS structures and their function in the pathogenesis of gastrointestinal disease. In this study, the presence of several LOS genes in 40 C. jejuni strains with different abilities to colonize the chicken gut and to invade Caco-2 cells was investigated by PCR. Two genes, cgtB and wlaN, encoding putative beta-1,3-galactosyltransferases were detected in most strongly invasive strains and rarely in non-invasive strains. A homopolymeric tract within the wlaN gene resulted in an intact gene product only in strongly invasive strains. The specific function of these genes during LOS biosynthesis is still unknown. cgtB and wlaN gene products are suggested to be involved in development of the colonization and invasion ability of C. jejuni. After a classification of the complete LOS loci, an association between a particular LOS class and colonization and invasion ability of the C. jejuni strain could not be detected. Lack of the pglB gene involved in protein glycosylation in one strain could be responsible for the weak colonization and invasion ability of this strain. There is some evidence that different genetic characteristics were responsible for strong or weak colonization and the invasion ability of C. jejuni strains. PMID:18033824

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

    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.

  19. Monomerization alters the dynamics of the lid region in Campylobacter jejuni CstII: an MD simulation study.

    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

  20. 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

    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.

  1. 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

    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

  2. Expression, purification, crystallization and preliminary X-ray characterization of two crystal forms of stationary-phase survival E protein from Campylobacter jejuni

    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.

  3. Campylobacter spp. Zoonotic micro organism

    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...

  4. Adaptation of Campylobacter jejuni to biocides used in the food industry affects biofilm structure, adhesion strength, and cross-resistance to clinical antimicrobial compounds.

    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

  5. Association of Anti-GT1a Antibodies with an Outbreak of Guillain-Barre Syndrome and Analysis of Ganglioside Mimicry in an Associated Campylobacter jejuni Strain.

    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.

  6. Exploiting the Campylobacter jejuni protein glycosylation system for glycoengineering vaccines and diagnostic tools directed against brucellosis

    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.

  7. Chitosan supplementation reduces enteric colonization of Campylobacter jejuni in broiler chickens and down-regulates expression of colonization genes

    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...

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

    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

  9. Genotyping of Campylobacter jejuni strains from Danish broiler chickens by restriction fragment length polymorphism of the LPS gene cluster

    Knudsen, K.N.; Bang, Dang Duong; Nielsen, E.M.;

    2005-01-01

    were 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...

  10. Validation according to ISO 16140:2003 of a commercial real-time PCR-based method for detecting Campylobacter jejuni, C. coli, and C. lari in foods.

    Vencia, W; Nogarol, C; Bianchi, D M; Gallina, S; Zuccon, F; Adriano, D; Gramaglia, M; Decastelli, L

    2014-05-01

    Campylobacteriosis was the most frequently reported zoonosis in the European Union (EU) in 2010, with Campylobacter jejuni, Campylobacter coli, and Campylobacter lari as the most frequently reported species in foodborne outbreaks (FBOs). Relatively sensitive to environmental factors, these species may be present in low numbers. In line with EU policy for food control and FBO detection and in view of the need to reduce response time, we validated an alternative molecular method according to ISO 16140:2003 which establishes the general principle and technical protocol for the validation of alternative methods in the microbiological analysis of food. We used a qualitative real-time PCR commercial kit for the detection of C. jejuni, C. coli, and C. lari in two food categories "fruit and vegetable-based products" and "dairy products". The validation protocol comprises two phases: the first is a method comparison study of the alternative method against the reference method, and the second is an interlaboratory study of each of the two methods. In the first step, ISO 16140:2003 validation examines the following parameters: limit of detection (LOD); relative accuracy, relative specificity and sensitivity; relative detection level (RDL); and inclusivity and exclusivity. Except for LOD, inclusivity and exclusivity, the other steps were performed against the reference method (ISO 10272:2006). The LOD of the real-time PCR method was set at 4CFU/25g or mL for both food categories. Relative accuracy (98.33%), specificity (96.77%), and sensitivity (100%) were recorded for the food category "fruit and vegetable-based products" and 93.3%, 88.24%, 100%, respectively, for "dairy products". The RDL according to Fisher's exact test was p=1 for both food categories, for each level, and each food/strain combination. The interlaboratory study results showed correct identification of all 24 blind samples with both methods by all the participating laboratories. The results show that this

  11. 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

    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

  12. It takes two to tango: two TatA paralogues and two redox enzyme-specific chaperones are involved in the localization of twin-arginine translocase substrates in Campylobacter jejuni.

    Liu, Y.W.; Hitchcock, A.; Salmon, R.C.; Kelly, D J

    2014-01-01

    The food-borne zoonotic pathogen Campylobacter jejuni has complex electron transport chains required for growth in the host, many of which contain cofactored periplasmic enzymes localized by the twin-arginine translocase (TAT). We report here the identification of two paralogues of the TatA translocase component in C. jejuni strain NCTC 11168, encoded by cj1176c (tatA1) and cj0786 (tatA2). Deletion mutants constructed in either or both of the tatA1 and tatA2 genes displayed distinct growth an...

  13. 空肠弯曲菌致病分子机制研究进展%Molecular Mechanisms on Pathogenesis of Campylobacter jejuni

    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.

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

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

    2004-01-01

    analysis of AFLP profiles derived from genomic DNA. Extensive genetic diversity was seen among the strains examined; however, 43 groups of isolates were identified at the 92% similarity (S-) level. Thirteen groups contained isolates from a single host, possibly representing genotypes of 'low risk' to human......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...... diarrhoea and diverse animal and environmental sources were collected mainly through a National surveillance programme in Denmark and serotyped by use of the established 'Penner' scheme. Genetic relationships among these isolates, and the archetypal serotype reference strains, were assessed by numerical...

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

    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.

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

    Krause-Gruszczynska, Malgorzata

    2011-12-28

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

  17. The combined efficacy of carvacrol and modified atmosphere packaging on the survival of Salmonella, Campylobacter jejuni and lactic acid bacteria on turkey breast cutlets.

    Nair, Divek V T; Kiess, Aaron; Nannapaneni, Rama; Schilling, Wes; Sharma, Chander Shekhar

    2015-08-01

    The primary objective of this study was to determine the efficacy of carvacrol in combination with modified atmosphere packaging (MAP) in reducing Salmonella on turkey breast cutlets stored at 4 °C. In experiment I, carvacrol (0.5, 1, and 2% v/v) was applied as surface treatment and samples were stored under aerobic condition or as surface and dip treatments followed by storage in an environment of 100% carbon dioxide. The findings of the experiment I revealed the synergistic activity of carvacrol with carbon dioxide in reducing Salmonella when used as dip treatment compared to the surface treatment. In experiment II, turkey breast cutlets were dip treated with carvacrol (0.25, 0.5, and 1% v/v) for 30 s and stored under MAP (95% carbon dioxide and 5% oxygen) to evaluate the efficacy against Salmonella, Campylobacter jejuni and lactic acid bacteria on turkey breast cutlets. In experiment II, the combined application of carvacrol and MAP resulted in 1.0-2.0 log CFU/g reduction (P ≤ 0.05) of both Salmonella and Campylobacter on turkey breast cutlets for 7 d storage at 4 °C. MAP alone and in combination with carvacrol reduced lactic acid bacteria (P ≤ 0.05) on cutlets stored at 4 °C for 21 d period. There was no difference (P ≤ 0.05) in meat color among treatments and controls except for an increased paleness of meat (P ≤ 0.05) observed for the 1% carvacrol treated cutlets stored under MAP after 21 d of storage. The high concentration of carbon dioxide and carvacrol treatments did not cause any alteration in meat pH (P ≤ 0.05). In conclusion, carvacrol was effective at a low concentration of 0.25% (v/v) in reducing Salmonella and C. jejuni by ∼1.0 log CFU/g when stored under MAP. PMID:25846923

  18. Evaluation of a real-time multiplex PCR for the simultaneous detection of Campylobacter jejuni, Salmonella spp., Shigella spp./EIEC, and Yersinia enterocolitica in fecal samples.

    Van Lint, P; De Witte, E; De Henau, H; De Muynck, A; Verstraeten, L; Van Herendael, B; Weekx, S

    2015-03-01

    Conventional diagnosis of infectious diarrhea caused by bacteria is time-consuming, labor-intensive, and has a suboptimal sensitivity. We have therefore developed a multiplex real-time polymerase chain reaction (PCR) for the simultaneous detection of Campylobacter jejuni, Salmonella spp., Shigella spp./enteroinvasive Escherichia coli (EIEC), and Yersinia enterocolitica in fecal samples. No cross reactivity between the different pathogens was observed, and the multiplex setup of the assay did not have an impact on the sensitivity of the PCR. The analytical sensitivity was 87 CFU/mL for C. jejuni, 61 CFU/mL for Shigella spp./EIEC, 5,528 CFU/mL for Salmonella spp., and 1,306 CFU/mL for Y. enterocolitica. An extensive validation of the assay was performed by testing 1,687 patient samples by both PCR and with conventional techniques. The use of PCR increased the overall clinical sensitivity from 78 to 100 % (p < 0.0001), the specificity was 99.4 % for the PCR, compared with 99.9 % for conventional culture. The novel PCR assay allows for rapid, sensitive, inexpensive, and high-throughput testing of the most common bacterial causes of gastroenteritis. PMID:25326870

  19. Campylobacter jejuni outer membrane vesicle-associated proteolytic activity promotes bacterial invasion by mediating cleavage of intestinal epithelial cell E-cadherin and occludin.

    Elmi, Abdi; Nasher, Fauzy; Jagatia, Heena; Gundogdu, Ozan; Bajaj-Elliott, Mona; Wren, Brendan; Dorrell, Nick

    2016-04-01

    Outer membrane vesicles (OMVs) play an important role in the pathogenicity of Gram-negative bacteria. Campylobacter jejuni produces OMVs that trigger IL-8, IL-6, hBD-3 and TNF-α responses from T84 intestinal epithelial cells and are cytotoxic to Caco-2 IECs and Galleria mellonella larvae. Proteomic analysis of 11168H OMVs identified the presence of three proteases, HtrA, Cj0511 and Cj1365c. In this study, 11168H OMVs were shown to possess proteolytic activity that was reduced by pretreatment with specific serine protease inhibitors. OMVs isolated from 11168H htrA, Cj0511 or Cj1365c mutants possess significantly reduced proteolytic activity. 11168H OMVs are able to cleave both E-cadherin and occludin, but this cleavage is reduced with OMVs pretreated with serine protease inhibitors and also with OMVs isolated from htrA or Cj1365c mutants. Co-incubation of T84 monolayers with 11168H OMVs results in a visible reduction in both E-cadherin and occludin. The addition of 11168H OMVs to the co-culture of live 11168H bacteria with T84 cells results in enhanced levels of bacterial adhesion and invasion in a time-dependent and dose-dependent manner. Further investigation of the cleavage of host cell structural proteins by C. jejuni OMVs should enhance our understanding of the interactions of this important pathogen with intestinal epithelial cells. PMID:26451973

  20. Phase variation of a Type IIG restriction-modification enzyme alters site-specific methylation patterns and gene expression in Campylobacter jejuni strain NCTC11168.

    Anjum, Awais; Brathwaite, Kelly J; Aidley, Jack; Connerton, Phillippa L; Cummings, Nicola J; Parkhill, Julian; Connerton, Ian; Bayliss, Christopher D

    2016-06-01

    Phase-variable restriction-modification systems are a feature of a diverse range of bacterial species. Stochastic, reversible switches in expression of the methyltransferase produces variation in methylation of specific sequences. Phase-variable methylation by both Type I and Type III methyltransferases is associated with altered gene expression and phenotypic variation. One phase-variable gene of Campylobacter jejuni encodes a homologue of an unusual Type IIG restriction-modification system in which the endonuclease and methyltransferase are encoded by a single gene. Using both inhibition of restriction and PacBio-derived methylome analyses of mutants and phase-variants, the cj0031c allele in C. jejuni strain NCTC11168 was demonstrated to specifically methylate adenine in 5'CCCGA and 5'CCTGA sequences. Alterations in the levels of specific transcripts were detected using RNA-Seq in phase-variants and mutants of cj0031c but these changes did not correlate with observed differences in phenotypic behaviour. Alterations in restriction of phage growth were also associated with phase variation (PV) of cj0031c and correlated with presence of sites in the genomes of these phages. We conclude that PV of a Type IIG restriction-modification system causes changes in site-specific methylation patterns and gene expression patterns that may indirectly change adaptive traits. PMID:26786317

  1. Effect of Different Feed Structures and Bedding on the Horizontal Spread of Campylobacter jejuni within Broiler Flocks

    Birgitte Moen

    2013-10-01

    Full Text Available In this study, we investigated the effects of different feed structures and beddings on the spread of C. jejuni in broiler flocks, and the effect on the cecal microbiota. Broiler chickens raised in 24 eight-bird group cages on either rubber mat or wood shavings were fed either a wheat-based control diet (Control, a diet where 50% of the ground wheat was replaced by whole wheat prior to pelleting (Wheat, or a wheat-based diet, such as the control diet diluted with 12% oat hulls (Oat. Samples from the cloacal mucosa of all birds were taken daily for C. jejuni quantification and cecum samples were collected at the end of the experiment for C. jejuni quantification and microbiota analyses. We have shown a statistically significant effect of increased feed structure on the reduced spread of C. jejuni in chicken flocks, but no significant differences were detected between types of structure included in the feed. No significant changes in the dominating microbiota in the lower lower gastrointestinal (GI tract were observed, which indicates that feed structure only has an effect on the upper GI tract. Delaying the spread of C. jejuni in broiler flocks could, at time of slaughter, result in fewer C. jejuni-positive broilers.

  2. 南京市2006年鸡肉产品中空肠弯曲菌污染状况调查%Investigation on status of campylobacter jejuni pollution in chicken products during 2006, Nanjing

    乔昕; 袁宝君; 吴高林

    2007-01-01

    空肠弯曲菌(campylobacter jejuni,CJ)1982年由Dekeyser等人首次从腹泻患者粪便中分离出来[1].近年来该菌感染率在世界各地普遍呈上升趋势,有的国家由其所致的急性腹泻甚至占95%[2].

  3. 空肠弯曲菌蛋白质组学的研究进展%Progress in Research on Proteomics of Campylobacter jejuni

    祝令伟

    2011-01-01

    Campylobacterjejuni is popular in the world, which is the leading cause of bacterial gastroenteritis and acute diarrhea in humans and animals.Proteomics encompasses the global analysis of proteins at the organism level.The technologies included under this term have now started to be utilized for understanding how Campylobacter species respond to changes in the environment,with an emphasis on the human host, as well as to map subcellular locations of proteins.This paper reviews the progress in research on proteomics of Campylobacter jejuni.%空肠弯曲菌病在世界各地均有发生,主要导致人和动物的细菌性胃肠炎及急性腹泻等.通过细菌蛋白质组学技术研究该类病原菌应对环境变化,尤其是应对宿主的反应机制,以及重要功能蛋白的亚细胞定位等,能够在蛋白水平深人研究弯曲菌病的致病机理.本文就空肠弯曲菌蛋白组学的研究进展作一综述.

  4. Healthy puppies and kittens as carriers of Campylobacter spp., with special reference to Campylobacter upsaliensis

    Hald, Birthe; Madsen, Mogens

    1997-01-01

    Living in a household with a dog or cat has previously been identified as a significant risk factor for acquiring campylobacteriosis, in particular, with reference to Campylobacter upsaliensis infection. In a cross-sectional study carried out in Denmark between August and December 1996, 72 healthy...... puppies and 42 healthy kittens, aged between 11 and 17 weeks, were sampled for fecal campylobacter shedding by culture of rectal swab specimens on blood-free agar base with cefoperazone at 32 mg/liter and amphotericin at 10 mg/liter and on blood-free agar base with cefoperazone at 8 mg/liter, teicoplanin...... Campylobacter spp., with a species distribution of 76% C. jejuni, 5% C. coli, and 19% C. upsaliensis, Of the kittens examined, two (5%) excreted campylobacters; both strains were C. upsaliensis, None of the chicken samples examined were found to be positive for C. upsaliensis. We concluded that young puppies...

  5. Study on antibacterial activity of garlic solution against Campylobacter jejuni%大蒜液对空肠弯曲杆菌的抑菌作用研究∗

    蒋小叶; 邱宏; 邓继家; 田瑶; 周秀萍

    2015-01-01

    Objective To explore the antibacterial activity of garlic solution against Campylobacter jejuni,and to provide theoriti-cal basis for further utilization of garlic.Methods Mashed garlic and extracted garlic solution.Antimicrobial susceptibility was de-termined by usig K-B disk diffusion test and broth dilution method.Results The minimal inhibitory concentration(MIC)of garlic solution against campylobacter jejuni was 40%.It showed positive correlation between bacteriostatic effect and concentration of gar-lic solution.Conclusion Garlic solution has strong bacteriostasis function to campylobacter jejuni.%目的:研究大蒜提取液对空肠弯曲杆菌的抑菌作用,为进一步利用大蒜资源提供理论依据。方法直接捣碎大蒜提取大蒜液,用琼脂纸片扩散法(K-B 纸片法)和肉汤稀释法检测大蒜液对空肠弯曲杆菌的抑菌作用。结果大蒜液对空肠弯曲杆菌的最小抑菌浓度(MIC)为40%,其抑菌活性与浓度呈正相关。结论大蒜液对空肠弯曲杆菌有较强的抑菌作用。

  6. Campylobacter jejuni dsb gene expression is regulated by iron in a Fur-dependent manner and by a translational coupling mechanism

    Grabowska Anna D

    2011-07-01

    Full Text Available Abstract Background Many bacterial extracytoplasmic proteins are stabilized by intramolecular disulfide bridges that are formed post-translationally between their cysteine residues. This protein modification plays an important role in bacterial pathogenesis, and is facilitated by the Dsb (disulfide bond family of the redox proteins. These proteins function in two parallel pathways in the periplasmic space: an oxidation pathway and an isomerization pathway. The Dsb oxidative pathway in Campylobacter jejuni is more complex than the one in the laboratory E. coli K-12 strain. Results In the C. jejuni 81-176 genome, the dsb genes of the oxidative pathway are arranged in three transcriptional units: dsbA2-dsbB-astA, dsbA1 and dba-dsbI. Their transcription responds to an environmental stimulus - iron availability - and is regulated in a Fur-dependent manner. Fur involvement in dsb gene regulation was proven by a reporter gene study in a C. jejuni wild type strain and its isogenic fur mutant. An electrophoretic mobility shift assay (EMSA confirmed that analyzed genes are members of the Fur regulon but each of them is regulated by a disparate mechanism, and both the iron-free and the iron-complexed Fur are able to bind in vitro to the C. jejuni promoter regions. This study led to identification of a new iron- and Fur-regulated promoter that drives dsbA1 gene expression in an indirect way. Moreover, the present work documents that synthesis of DsbI oxidoreductase is controlled by the mechanism of translational coupling. The importance of a secondary dba-dsbI mRNA structure for dsbI mRNA translation was verified by estimating individual dsbI gene expression from its own promoter. Conclusions The present work shows that iron concentration is a significant factor in dsb gene transcription. These results support the concept that iron concentration - also through its influence on dsb gene expression - might control the abundance of extracytoplasmic proteins

  7. Campylobacter fetus Bacteremia Revealed by Cellulitis without Gastrointestinal Symptoms in the Context of Acquired Hypogammaglobulinemia: A Report of Three Cases

    Souleymane Brah

    2011-01-01

    Full Text Available Campylobacter fetus bacteremia is rare and occurs mainly in patients with immunosuppression. This infection, which often involves secondary localizations has already been reported in some primary humoral immune deficiencies. We describe three cases of severe infection due to C. fetus with cellulitis at presentation, but without any gastrointestinal symptoms, occurring in patients with acquired hypogammaglobulinemia.

  8. Decreased competiveness of the foodborne pathogen, Campylobacter jejuni, co-culture with the hyper-ammonia anaerobe, Clostridium aminophilum

    Campylobacter spp. are a leading bacterial cause of human foodborne illness. When co-cultured in anaerobic Bolton broth with the hyper-ammonia-producing bacterium, Clostridium aminophilum, ammonia accumulation was greater (P 1...

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

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

    2010-01-01

    inoculated in brain heart infusion broth containing 0.3% tartaric acid. On chicken meat medallions, reductions of C. jejuni were 0.5 to 2 log units when tartaric acid solutions (2, 4, 6, and 10%) were spread onto the meat. Analysis of acidic food ingredient (e.g., vinegar, lemon juice, pomegranate syrup......, and soya sauce) revealed that such ingredients reduced counts of C. jejuni by at least 0.8 log units on meat medallions. Three low pH marinades (pH syrup, lemon juice, and white wine vinegar were prepared. When applied to whole filets, these marinades resulted in a reduction...

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

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

    2014-01-01

    assays demonstrated that phage F341 does not adsorb to these nonmotile C. jejuni NCTC11168 mutants. Taken together, we propose that phage F341 uses the flagellum as a receptor. Phage-host interactions were investigated using fluorescence confocal and transmission electron microscopy. These data...... demonstrate that F341 binds to the flagellum by perpendicular attachment with visible phage tail fibers interacting directly with the flagellum. Our data are consistent with the movement of the C. jejuni flagellum being required for F341 to travel along the filament to reach the basal body of the bacterium...

  11. 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

    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。

  12. Optimisation of the PCR Method for the Detection of Campylobacter jejuni and Campylobacter coli in Samples of Ready-to-Eat Chicken Meals

    Maděrová, Zdeňka; Demnerová, K.; Pazlarová, J.

    2008-01-01

    Roč. 26, - (2008), s. 291-297. ISSN 1212-1800 Institutional research plan: CEZ:AV0Z60870520 Keywords : polymerase chain reaction * internal control * Campylobacter spp. Subject RIV: EE - Microbiology, Virology Impact factor: 0.472, year: 2008

  13. Fluoroquinolone resistance in Campylobacter

    Fluoroquinolone-resistant Campylobacter jejuni and C. coli are common in animals because of the use of fluoroquinolones as therapeutic agents in animal husbandry, particularly in chickens and other poultry. Campylobacter is a commensal in poultry, and therefore, poultry and poultry products are the...

  14. Identification des facteurs environnementaux responsables de la presence de Campylobacter Jejuni dans les eaux de surface de l'Estrie (Quebec)

    Bonfils, Djoan

    Background: The sources and the epidemiology of Campylobacter jejuni (CJ) in environmental water are not well understood. We developed a spatial analysis methodology able to identify the main environmental factors predicting the presence and quantity of Campylobacter in river water. Methods: For two years, water samples were collected weekly and scanned for CJ and fecal coliforms at 32 sampling sites of Estrie hydrographic network, Québec. The quantity of CJ in water was estimated using the Most Probable Number method (MPN). The 32 sampling sites were linked to their catchment area; 10 of them were excluded from the analysis, because their hydrographic basin was not independent from the other sites. For each site, the following environmental variables were included: Water flow, slope, land-cover, land-use including type of farming, animal density, total precipitation in the 3 days prior to water sampling. A stepwise multivariate regression was realized across the different analysis windows to define the size of the area upstream from the sampling sites (from 1.5 to 24 km) which was the most closely associated with the mean quantity of CJ, and which environmental factors were associated with a higher mean quantity of CJ in water. Results: Preliminary results show that an area defined by a radius of 14 km upstream of the sample site was the most contributing zone for the bacteria (r2=0.38, p=0.002). Within this 14 km area, the only significant variable associated with a higher mean quantity of CJ was bovine density (p=0.002). When analyzing the data within a 120 m buffer zone across 14 km upstream of the sampling sites (r2=0.40, p=0.001), the only significant variable associated with a higher mean quantity of CJ was the percentage of agricultural surface (p=0.001). Conclusions: These results suggest a strong implication of bovine density in conjunction with crops and associated manure spreading on the quantity of CJ in environmental water. Keywords: Campylobacter

  15. 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

    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

  16. 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

    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

  17. Identification of DNA sequence variation in Campylobacter jejuni strains associated with the Guillain-Barré syndrome by high-throughput AFLP analysis

    Endtz Hubert P

    2006-04-01

    Full Text Available Abstract Background Campylobacter jejuni is the predominant cause of antecedent infection in post-infectious neuropathies such as the Guillain-Barré (GBS and Miller Fisher syndromes (MFS. GBS and MFS are probably induced by molecular mimicry between human gangliosides and bacterial lipo-oligosaccharides (LOS. This study describes a new C. jejuni-specific high-throughput AFLP (htAFLP approach for detection and identification of DNA polymorphism, in general, and of putative GBS/MFS-markers, in particular. Results We compared 6 different isolates of the "genome strain" NCTC 11168 obtained from different laboratories. HtAFLP analysis generated approximately 3000 markers per stain, 19 of which were polymorphic. The DNA polymorphisms could not be confirmed by PCR-RFLP analysis, suggesting a baseline level of 0.6% AFLP artefacts. Comparison of NCTC 11168 with 4 GBS-associated strains revealed 23 potentially GBS-specific markers, 17 of which were identified by DNA sequencing. A collection of 27 GBS/MFS-associated and 17 enteritis control strains was analyzed with PCR-RFLP tests based on 11 of these markers. We identified 3 markers, located in the LOS biosynthesis genes cj1136, cj1138 and cj1139c, that were significantly associated with GBS (P = 0.024, P = 0.047 and P Conclusion This study shows that bacterial GBS markers are limited in number and located in the LOS biosynthesis genes, which corroborates the current consensus that LOS mimicry may be the prime etiologic determinant of GBS. Furthermore, our results demonstrate that htAFLP, with its high reproducibility and resolution, is an effective technique for the detection and subsequent identification of putative bacterial disease markers.

  18. Dual Repression of the Multidrug Efflux Pump CmeABC by CosR and CmeR in Campylobacter jejuni

    Grinnage-Pulley, Tara; Mu, Yang; Dai, Lei; Zhang, Qijing

    2016-01-01

    During transmission and intestinal colonization, Campylobacter jejuni, a major foodborne human pathogen, experiences oxidative stress. CosR, a response regulator in C. jejuni, modulates the oxidative stress response and represses expression of the CmeABC multidrug efflux pump. CmeABC, a key component in resistance to toxic compounds including antimicrobials and bile salts, is also under negative regulation by CmeR, a TetR family transcriptional regulator. How CosR and CmeR interact in binding to the cmeABC promoter and how CosR senses oxidative stress are still unknown. To answer these questions, we conducted various experiments utilizing electrophoretic mobility shift assays and transcriptional fusion assays. CosR and CmeR bound independently to two separate sites of the cmeABC promoter, simultaneously repressing cmeABC expression. This dual binding of CosR and CmeR is optimal with a 17 base pair space between the two binding sites as mutations that shortened the distance between the binding sites decreased binding by CmeR and enhanced cmeABC expression. Additionally, the single cysteine residue (C218) of CosR was sensitive to oxidation, which altered the DNA-binding activity of CosR and dissociated CosR from the cmeABC promoter as determined by electrophoretic mobility shift assay. Replacement of C218 with serine rendered CosR insensitive to oxidation, suggesting a potential role of C218 in sensing oxidative stress and providing a possible mechanism for CosR-mediated response to oxidative stress. These findings reveal a dual regulatory role of CosR and CmeR in modulating cmeABC expression and suggest a potential mechanism that may explain overexpression of cmeABC in response to oxidative stress. Differential expression of cmeABC mediated by CmeR and CosR in response to different signals may facilitate adaptation of Campylobacter to various environmental conditions. PMID:27468281

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

    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...

  20. Draft Genome Sequence of Lactobacillus crispatus JCM5810, Which Can Reduce Campylobacter jejuni Colonization in Chicken Intestine.

    Wooten, Jessica; Liu, Xiaoji; Miller, Michael J

    2016-01-01

    We present the 2.05-Mb draft genome sequence ofLactobacillus crispatusJCM5810, a chicken intestinal isolate with the ability to reduceCampylobacter jejunicolonization in chickens. The genome sequence will provide insights on the probiotic mechanisms ofL. crispatusJCM5810. PMID:27081134

  1. 食品中空肠弯曲菌检测技术最新进展%Progress on detection methods of Campylobacter jejuni in foods

    胡元庆; 李凤霞

    2014-01-01

    空肠弯曲菌(Campylobacter jejuni)是人类常见的食源性病原菌,感染后主要引起急性肠炎,并且与格林-巴利综合症密切相关.近年来弯曲菌污染禽肉及牛奶制品引发的食品安全事件呈上升趋势,发达国家尤为突出.高效快速检测动物性食品的加工、贮藏和消费过程中弯曲菌污染情况十分必要,可为有效控制该菌污染食品及人弯曲菌病发生提供技术保障.本文综述了近年来国内外关于食品加工及贮藏过程中弯曲菌污染检测技术的最新成果.

  2. Characterization of CetA and CetB, a bipartite energy taxis system in Campylobacter jejuni

    Elliott, Kathryn T; DiRita, Victor J.

    2008-01-01

    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 jej...

  3. Molecular subtyping methods for campylobacter

    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. Comparative genome ind...

  4. HIV/AIDS相关性慢性腹泻患者感染空肠弯曲菌的临床分析%Infections of Campylobacter Jejuni among Patients with HIV/AIDS-related Chronic Diarrhea

    华文浩; 王慧珠; 赵辉; 李敏; 张燕; 田敬华; 王玉光

    2011-01-01

    Objective To investigate the Campylobacter jejuni infection status of HIV/AIDS patients with chronic diarrhea in Beijing, Henan, and Xinjiang.Methods Micro - aerobic cultivation of stool specimens were carried out in 253 cases of HIV/AIDS patients with chronic diarrhea in Beijing, Henan, and Xinjiang.The colonies were identified according to morphology, Gram staining, and results frombiochemical reactions.Results Two strains of Campylobacter jejuni were identified from the 253 cases of HIV/AIDS - related chronic diarrhea ( infection rate: 0.8% ).Conclusion It is important that monitoring for Campylobacter jejuni be carried out in high risk patients for prevention and control of the infection of the bacteria.%目的 了解北京、河南、新疆三地HIV/AIDS相关性慢性腹泻患者空肠弯曲菌的感染状况.方法 对北京、河南、新疆三地253例HIV/AIDS慢性腹泻患者的粪便标本进行微需氧培养,根据菌落的形态、菌体的革兰染色及生化反应进行鉴定.结果 从253例HIV/AIDS相关性慢性腹泻患者的粪便中共检出2株空肠弯曲菌,感染率为0.8%.结论 加强空肠弯曲菌的监测是预防和控制空肠弯曲菌病的重要手段.

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

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

    2012-01-01

    . Typhimurium over the time course of 15 days. In contrast, the number of Cercomonas sp. cells decreased when grown with or without L. monocytogenes for 9 days of co-culture. Interestingly, we observed that C. jejuni and S. Typhimurium survived better when co-cultured with flagellates than when cultured alone....... The results of this study suggest that Cercomonas sp. and perhaps other soil flagellates may play a role for the survival of food-borne pathogens on plant surfaces and in soil....

  6. 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

    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。

  7. Cloning and alignment of WaaF gene of Campylobacter jejuni Lulei%空肠弯曲菌Lulei株WaaF基因克隆与序列分析

    邢丛丛; 徐飞; 白欣立; 付金生; 李文倩; 卢晓卫; 马海阳; 李震中

    2012-01-01

    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-Barrg 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.%目的 克隆空肠弯曲菌Lulei株WaaF基因,并分析其遗传进化关系.方法 聚合酶链反应扩增空肠弯曲菌Lulei株WaaF基因,构建pGEM-T-WaaF质粒,选择正确克隆质粒测序.自美国国家生物技术信息中心Nucleotide数据库下载5株吉兰-巴雷综合征相关菌株WaaF序列,1株非吉兰-巴雷综合征相关菌株形WaaF序列,利用DNAstar7.1软件建立进化树,分析不同菌株间WaaF基因遗传关系.结果 完成了空肠弯曲菌Lulei株WaaF基因的克隆,发现WaaF基因在吉兰-巴雷综合征相关菌株间存在聚类现象.结论 空肠弯曲菌Lulei株WaaF基因是807 bp的片段,与Lichang株遗传进化关系最近.

  8. Campylobacter jejuni induces an anti-inflammatory response in human intestinal epithelial cells through activation of phosphatidylinositol 3-kinase/Akt pathway

    Li, Yiping; Vegge, Christina S.; Brøndsted, Lone;

    2011-01-01

    Campylobacterjejuni (C. jejuni) is the most common cause of human acute bacterial gastroenteritis. Poultry is a major reservoir of C. jejuni and considered an important source of human infections, thus, it is important to understand the host response to C. jejuni from chicken origin. In this study...

  9. Detecção dos genes codificantes da toxina CDT, e pesquisa de fatores que influenciam na produção de hemolisinas em amostras de Campylobacter jejuni de origem avícola

    Michele M. Trindade

    2015-08-01

    Full Text Available Resumo: Membros termofílicos do gênero Campylobacter são reconhecidos como importantes enteropatógenos para o ser humano e animais. A grande diversidade ecológica destes micro-organismos em diferentes habitats tais como água, animais e alimentos predispõem ao aparecimento de novos fatores de virulência. Este trabalho teve por objetivo detectar os genes codificantes da Toxina Distensiva Citoletal (CDT por meio da técnica de PCR, pesquisar a atividade de hemolisinas e a influência de soluções quelantes e de íons nesta atividade. Foram utilizadas 45 amostras de Campylobacter jejuni de origem avícola para pesquisa de atividade hemolítica, cultivadas em Caldo Triptona de Soja (TSB. Após o crescimento bacteriano, as amostras foram semeadas em Ágar tríptico de soja (TSA contendo 5% de sangue de ovino. Para verificar a influência de agentes quelantes e solução de íons na atividade hemolítica, as amostras de C. jejuni foram cultivadas em TSB contendo separadamente os quelantes EDTA, ácido acético, soluções de íons CaCl2, MgCl2 e FeCl3, em atmosfera de microaerofilia. Quanto à atividade de hemolisina de C. jejuni em placas de TSA - sangue ovino foi possível observar que houve hemólise em 40% das amostras analisadas apenas com caldo TSB. Somente o ácido acético apresentou ação quelante sobre a atividade de hemolisinas em amostras de C. jejuni semeadas em placas de TSA - sangue ovino. Para detecção dos genes cdtA, cdtB e cdtC através da técnica da Reação em Cadeia da Polimerase (PCR foram utilizadas 119 amostras de C. jejuni de origem avícola. Foi possível observar que 37,8% possuíam o perfil de genes cdtABC. Os resultados demonstraram em amostras avícolas a presença de cepas de C. jejuni com potencial virulento, devido à presença dos genes da toxina CDT e potencial hemolítico, que apresentou ação reduzida in vitro com ácido acético.

  10. A Polydiacetylene Vesicle-based Colorimetric Detection of Campylobacter jejuni Using a DNA Aptamer as Recognition Element%聚丁二炔纳米囊泡生物传感器比色检测空肠弯曲菌的初步研究

    董建涛; 欧阳后先; 徐柯; 吴文鹤; 吕建新

    2012-01-01

    A polydiacetylene vesicle-based biosensor for rapid colorimetric detection of Campylobacter jejuni was developed in this work. Aptamers coupled to the surface of polydiacetylene nanovesicle by chemical reaction in order to achieve vesicle functionalization. Molecular recognition between Campylobacter jejuni and aptamers on the surface of the vesicle lead to absorption spectrum transition of polydiacetylene by which quantitative detection of Campylobacter jejuni could be realized. This method can detect Campylobacter jejuni in 30 mins with high specificity and simple operation.%本实验构建一种可以快速比色检测空肠弯曲菌的聚丁二炔纳米囊泡生物传感器.适配体通过化学反应耦合到聚丁二炔纳米囊泡表面,完成对囊泡的功能化.适配体与空肠弯曲菌特异识别,引起纳米囊泡溶液吸收光谱的改变,通过比色响应值衡量检测体系中有无空肠弯曲菌.该方法特异性好,与PCR对比实验相符;操作简便,检测时间短,仅需30 min.

  11. 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

    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

  12. 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

    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.

  13. 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

    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

  14. Construction and Characterization of Campylobacter jejuni flhA Mutant Strain%空肠弯曲菌flhA突变株的构建及其功能研究

    惠星星; 李烨; 王小元

    2011-01-01

    flhA, encoded by the gene flhA, is an important component of the flagellar export apparatus of Campylobacter jejuni. Here we constructed an flhA mutant strain HXW001.Compared with that of the wild type, the flhA mutant cell could grow well but lack flagella, the motility of the mutant cells was lost, and the ability of the mutant cells for the formation of biofilm and autoagglutination also decreased. In addition, it was found that the expression of the flagellin gene flaA was affected by flhA. These findings indicated that flhA was the molecular basis of flagellar synthesis and motility of Campylobacter jejuni and that flhA was closely related to pathogenicity of Carnpylobacter jejuni.%flhA是空肠弯曲茵鞭毛输出装置的关键基因,通过构建Campylobacterjejuni flhA突变株HXW001,进行毒力相关表型研究和分析.结果显示HXW001生长性能稳定,但其鞭毛和运动能力丧失,生物膜形成和自身凝集现象明显下降.RT-PCR实验证明flhA对鞭毛丝主要结构蛋白基因flaA的表达有一定的调控功能.研究表明flhA是空肠弯曲茵鞭毛生成和运动能力重要的分子基础,与空肠弯曲菌致病性密切有关.

  15. Multicenter evaluation of the BD max enteric bacterial panel PCR assay for rapid detection of Salmonella spp., Shigella spp., Campylobacter spp. (C. jejuni and C. coli), and Shiga toxin 1 and 2 genes.

    Harrington, S M; Buchan, B W; Doern, C; Fader, R; Ferraro, M J; Pillai, D R; Rychert, J; Doyle, L; Lainesse, A; Karchmer, T; Mortensen, J E

    2015-05-01

    Diarrhea due to enteric bacterial pathogens causes significant morbidity and mortality in the United States and worldwide. However, bacterial pathogens may be infrequently identified. Currently, culture and enzyme immunoassays (EIAs) are the primary methods used by clinical laboratories to detect enteric bacterial pathogens. We conducted a multicenter evaluation of the BD Max enteric bacterial panel (EBP) PCR assay in comparison to culture for the detection of Salmonella spp., Shigella spp., Campylobacter jejuni, and Campylobacter coli and an EIA for Shiga toxins 1 and 2. A total of 4,242 preserved or unpreserved stool specimens, including 3,457 specimens collected prospectively and 785 frozen, retrospective samples, were evaluated. Compared to culture or EIA, the positive percent agreement (PPA) and negative percent agreement (NPA) values for the BD Max EBP assay for all specimens combined were as follows: 97.1% and 99.2% for Salmonella spp., 99.1% and 99.7% for Shigella spp., 97.2% and 98.4% for C. jejuni and C. coli, and 97.4% and 99.3% for Shiga toxins, respectively. Discrepant results for prospective samples were resolved with alternate PCR assays and bidirectional sequencing of amplicons. Following discrepant analysis, PPA and NPA values were as follows: 97.3% and 99.8% for Salmonella spp., 99.2% and 100% for Shigella spp., 97.5% and 99.0% for C. jejuni and C. coli, and 100% and 99.7% for Shiga toxins, respectively. No differences in detection were observed for samples preserved in Cary-Blair medium and unpreserved samples. In this large, multicenter study, the BD Max EBP assay showed superior sensitivity compared to conventional methods and excellent specificity for the detection of enteric bacterial pathogens in stool specimens. PMID:25740779

  16. EAN中神经抗原P2蛋白有效表位的鉴定及其与空肠弯曲菌LPS抗原性比较%Identification of dominant epitope of P2 in EAN and studying of similarity betwe en the dominant epitope and LPS of Campylobacter jejuni

    李杰; 孙世杰; 于春雷; 王莉; 李一

    2001-01-01

    Objective:Identification of dominant epitope o f P2 in EAN and studying of similarity between the dominant epitope and LPS of Cam p ylobacter jejuni.Methods:Screening peptide library with EA N serum.Identified positive clone by ELISA and compared similarity between positi ve clone and LPS of Campylobacter jejuni.Results:Affinity of all indentified positive clone increas ed remarkbly compared to negative control(P<0.01),suggested they might be s pecific clones.The affinity between EAN serum and LPS increased remarkably compa red to negative control(P<0.01),but similar with positive clones.Conclusion:The result indicated that specific phage clones which have similar epitope with P2 peptide can be acquired by phage display tec h nique;Epitope mimicry may be identified between LPS of Campylobacter jejuni and auto-neural antigen,LPS may be an important factor in inducing auto-reactive i mmu ne response.Epitope mimicry mechanism is a possible mechanism for instigating an autoimmune response to self antigens that in turm can lead to autoimmune diseas e GBS.%目的:EAN中神经抗原(P2蛋白)的有效表位的鉴定以及将其与 空肠弯曲菌(Campylobacter jejuni,Cj)LPS抗原性比较。方法:采用噬菌体呈现技术对8个EAN模型血清抗体进行肽库筛选,并将 获得的克隆用ELISA方法进行阳性鉴定,以及将阳性克隆与空肠弯曲菌LPS相似性进行比较。结果:33个阳性克隆的亲和性均显著高于阴性对照(P<0.01),说 明它们是具有特异性的克隆;C.J.LPS与血清的亲和力显著高于对照组(P<0.01), 与阳性克隆的OD值相差不显著,C.J.LPS与阳性克隆之间存在相似性。结论:噬菌体呈现技术可筛选出与P2蛋白具有相同表位的噬菌体克隆; 空肠 弯曲菌LPS与P2蛋白之间存在表位模拟,C.J.LPS可能是启动GBS自身反应性免疫应答的重 要因素。

  17. The role of radiology in Campylobacter enterocolitis

    Demaerel, P.; Ponette, E.; Baert, A.L. (Leuven Univ. (Belgium). Dept. of Radiology); Lacquet, F.; Verbist, L. (Leuven Univ. (Belgium). Dept. of Microbiology); Rutgeerts, A.L. (Leuven Univ. (Belgium). Dept. of Internal Medicine)

    1989-05-01

    A series of 18 patients with diarrhoea and positive stool cultures for Campylobacter jejuni is presented. The most important radiological features were thickening of ileal mucosal folds, of interhaustral indentations and of the ileocaecal valve, lymphoid hyperplasia and microulcerations. Radiology, as well as endoscopy, are both non-specific in Campylobacter jejuni enterocolitis. The importance of radiology is to exclude more typical features of other causes of inflammatory bowel diseases. Moreover, before the result of the stool culture is available, the radiological features should suggest the suspicion of an acute infectious enterocolitis by Campylobacter jejuni as possible diagnosis. (orig.).

  18. Rapid Detection of Campylobacter jejuni by Real-Time PCR%实时荧光PCR方法快速检测空肠弯曲菌方法的建立

    刘俊华; 张欣强; 陈守义; 高秀洁; 李杰

    2009-01-01

    Objective To establish a real-time polymerase chain reaction (real-time PCR) assay for rapid detection of Campylobacter jejuni. Methods The special sequence of HipO gene of Campylobacter jejuni was amplified and characterized with a pair primers and TaqMan probe. Campylobacter coli, Salmonella, Vibrio cholerae and some other foedbome pathogens were used as specificity reference for the method. By adding standard culture fluid in blank fecal sample, the sensitivity of the method was evaluated. Results The real-time PCR method was designed to sensitively detect and identify C. jejuni from C. coli and other food-borne pathogens. The total assay could be completed in 80 min with a detection limit of approximated 5 CFU. And by adding C.jejuni culture fluid in blank fecal sample, 10~100 CFU could be detected with the RT-PCR method. Conclusion Result indicated that RT-PCR detection methods provide not only a special, sensitive, rapid, reproducible and accurate method for quantitative detection of C. jejuni, but also an important method for researching pathogenesis of C. jejuni.%目的 建立实时荧光PCR快速检测空肠弯曲菌的方法.方法 以空肠弯曲杆菌HipO基因的保守序列为模板设计特异性引物探针,建立一种能快速检测样本中空肠弯曲杆菌的实时荧光PCR方法:对方法的特异性和敏感性进行评价,并以正常人粪便为空白样本,添加一定量空肠弯曲菌标准株菌液进行检测,以对方法的检测效果进行初步评价.结果 该实时荧光PCR方法只对空肠弯曲杆菌进行特异扩增,同种属的结肠弯曲菌及其他常见食源性病原菌均不能扩增;整个检测过程只需要80 min,对空肠弯曲菌菌悬液可检测至5个细菌,对加标粪便样本可检测至10~100个细菌.结论 本研究建立的实时荧光PCR检测空肠弯曲菌方法不仅能实现对空弯菌的快速检测,而且还为空弯菌的快速诊断及其引起的食源性疾病的监控溯源提供有意义的参考.

  19. Structure and Active Stie Residues of Pg1D, an N-Acetyltransferase from the Bacillosamine Synthetic Pathway Required for N-Glycan Synthesis in Campylobacter jejuni

    Rangarajan,E.; Ruane, K.; Sulea, T.; Watson, D.; Proteau, A.; Leclerc, S.; Cygler, M.; Matte, A.; Young, N.

    2008-01-01

    Campylobacter jejuni is highly unusual among bacteria in forming N-linked glycoproteins. The heptasaccharide produced by its pgl system is attached to protein Asn through its terminal 2, 4-diacetamido-2, 4,6-trideoxy-d-Glc (QuiNAc4NAc or N, N'-diacetylbacillosamine) moiety. The crucial, last part of this sugar's synthesis is the acetylation of UDP-2-acetamido-4-amino-2, 4,6-trideoxy-d-Glc by the enzyme PglD, with acetyl-CoA as a cosubstrate. We have determined the crystal structures of PglD in CoA-bound and unbound forms, refined to 1.8 and 1.75 Angstroms resolution, respectively. PglD is a trimer of subunits each comprised of two domains, an N-terminal {alpha}/{beta}-domain and a C-terminal left-handed {beta}-helix. Few structural differences accompany CoA binding, except in the C-terminal region following the {beta}-helix (residues 189-195), which adopts an extended structure in the unbound form and folds to extend the {beta}-helix upon binding CoA. Computational molecular docking suggests a different mode of nucleotide-sugar binding with respect to the acetyl-CoA donor, with the molecules arranged in an 'L-shape', compared with the 'in-line' orientation in related enzymes. Modeling indicates that the oxyanion intermediate would be stabilized by the NH group of Gly143', with His125' the most likely residue to function as a general base, removing H+ from the amino group prior to nucleophilic attack at the carbonyl carbon of acetyl-CoA. Site-specific mutations of active site residues confirmed the importance of His125', Glu124', and Asn118. We conclude that Asn118 exerts its function by stabilizing the intricate hydrogen bonding network within the active site and that Glu124' may function to increase the pKa of the putative general base, His125'.

  20. Fold-recognition and comparative modeling of human α2,3-sialyltransferases reveal their sequence and structural similarities to CstII from Campylobacter jejuni

    Balaji Petety V

    2006-04-01

    Full Text Available Abstract Background The 3-D structure of none of the eukaryotic sialyltransferases (SiaTs has been determined so far. Sequence alignment algorithms such as BLAST and PSI-BLAST could not detect a homolog of these enzymes from the protein databank. SiaTs, thus, belong to the hard/medium target category in the CASP experiments. The objective of the current work is to model the 3-D structures of human SiaTs which transfer the sialic acid in α2,3-linkage viz., ST3Gal I, II, III, IV, V, and VI, using fold-recognition and comparative modeling methods. The pair-wise sequence similarity among these six enzymes ranges from 41 to 63%. Results Unlike the sequence similarity servers, fold-recognition servers identified CstII, a α2,3/8 dual-activity SiaT from Campylobacter jejuni as the homolog of all the six ST3Gals; the level of sequence similarity between CstII and ST3Gals is only 15–20% and the similarity is restricted to well-characterized motif regions of ST3Gals. Deriving template-target sequence alignments for the entire ST3Gal sequence was not straightforward: the fold-recognition servers could not find a template for the region preceding the L-motif and that between the L- and S-motifs. Multiple structural templates were identified to model these regions and template identification-modeling-evaluation had to be performed iteratively to choose the most appropriate templates. The modeled structures have acceptable stereochemical properties and are also able to provide qualitative rationalizations for some of the site-directed mutagenesis results reported in literature. Apart from the predicted models, an unexpected but valuable finding from this study is the sequential and structural relatedness of family GT42 and family GT29 SiaTs. Conclusion The modeled 3-D structures can be used for docking and other modeling studies and for the rational identification of residues to be mutated to impart desired properties such as altered stability, substrate

  1. 大蒜素对空肠弯曲杆菌抑菌作用的研究%Study on anti-microbial activity of allicin on campylobacter jejuni

    潘洪涛; 杨宗兴; 赵文萃; 王艳萍; 宋海艳; 张琦

    2015-01-01

    目的:研究大蒜素对空肠弯曲杆菌的抑菌作用和强度。方法采用纸片扩散法(K‐B法)测定抑菌圈直径;采用肉汤稀释法分别测定大蒜素、环丙沙星、红霉素对空肠弯曲杆菌的最低抑菌浓度(M IC ),比较大蒜素、环丙沙星、红霉素的最低抑菌浓度及抑菌率。结果大蒜素(M IC为12.80μg/m l ,抑菌率90.40%)对空肠弯曲杆菌的抑菌作用较环丙沙星(M IC为20.48μg/m l ,抑菌率90.21%)和红霉素(M IC为35.84μg/m l ,抑菌率90.33%)更强。结论大蒜素对空肠弯曲杆菌具有抑菌作用。%Objective To study the anti‐microbial activity and strength of allicin on Campylobacter jejuni .Methods Disc diffusion method (K‐B) was used to determine the diameter of the bacteriostatic ring .The minimal inhibitory concentra‐tion (MIC) of allicin ,ciprofloxacin and erythromycin were detected by constant broth dilution method ,respectively .The mini‐mal inhibitory concentrations and bacteriostatic rate of allicin ,ciprofloxacin and erythromycin were compared .Results The anti‐microbial activity on Campylobacter jejuni of allicin (MIC 12 .8 μg/ml ,bactetiostatic rate 90 .40% ) was better than that of ciprofloxacin (MIC 20 .48 μg/ml ,bactetiostatic rate 90 .21% ) and erythromycin (MIC 35 .84 μg/ml ,bactetiostatic rate 90 . 33% ) .Conclusion Allicin has anti‐microbial activity on Campylobacter jejuni in vitro .

  2. Interaction of campylobacter species with antibody, complement and phagocytes.

    Bernatowska, E.; Jose, P; Davies, H; Stephenson, M.; Webster, D

    1989-01-01

    The opsonisation of four different campylobacter species for human neutrophils was studied using a chemiluminescence system and electron microscopy. Opsonisation of Campylobacter fetus, Campylobacter coli, and Campylobacter jejuni was mediated by antibody and enhanced by complement. Antibody was not, however, required for the phagocytosis of Campylobacter pylori because it activates the classical pathway of complement directly. This unusual property may be important in the pathogenesis of C p...

  3. Study on the Relationship between Campylobacter Jejuni Infection and Guillain-Barre Syndrome%空肠弯曲菌感染与格林--巴利综合征的关系研究

    陈晶晶; 李春岩; 张建中; 刘瑞春; 田文强

    1995-01-01

    应用聚丙烯酰胺凝胶电泳(SDS-PAGE)技术,免疫印迹分析和酶联免疫斑点检测方法,对不同型格林--巴利综合征(Guillain-Barre,Syndrome,GBS)患者的神经及从病人中分离的空肠弯曲菌(Campylobacter jejuni,G.J)进行了血清免疫反应分析,结果表明空肠弯曲菌与经典型GBS的关系比该病的亚型急性运动性轴索性神经病(Acute Motor Axonal New Neuropa-thy,AMAN)更为密切.提示空肠弯曲菌感染可能是格林-巴利综合征的重要诱发因素之一.

  4. Analysis on monitoring results of Campylobacter jejuni in Nantong by using multiple PCR technology%多重PCR技术对南通市空肠弯曲菌监测结果分析

    熊海平; 苏婧; 黄红玫; 许海燕

    2014-01-01

    Objective:To understand the distribution of Campylobacter jejuni from chickens, poultry meat, the popu-lation and the environment in our city. Methods:Multiple PCR method was applied to detect Campylobacter jejuni from anal swabs of adult chicken, chicken manure, environmental samples of chicken farms, fresh poultry meat, frozen chicken, environmental samples of open fair, feces of diarrhea patients and anal swabs of healthy physical examination. All the data were processed and analyzed by SPSS 19.0. Results:There were 38 positive specimens(31.93%) in 119 anal swabs of adult chicken, 9 (7.26%) positive in 124 chicken manure, 5 (8.93%) positive in 56 environmental samples of chicken farms, 28 (17.28%) positive in 162 fresh poultry meat, 2(1.12%) positive in 178 frozen chicken, and 4(7.02%) positive in 57 envi-ronmental samples of open fair, 9 (2.15%) positive in 418 diarrhea cases and none positive in healthy physical examina-tion. Conclusion:Except for healthy physical examination, Campylobacter jejuni were detected in 7 categories of samples, which prompts the city to consider the risk of Campylobacter jejuni diseases, and to strengthen control measures.%目的:了解南通市不同鸡群、鸡肉、人群及环境中空肠弯曲菌的分布状况。方法:采用多重PCR法对相关鸡的样品、农贸市场环境样品、腹泻患者粪便及健康体检人员肛拭子等进行空肠弯曲菌检测。结果:119份成年鸡肛拭子、124份苗鸡粪便和56份鸡养殖场环境样品分别检出38份、9份和5份空肠弯曲菌阳性,阳性率分别为31.93%、7.26%和8.93%;162份新鲜鸡肉、178份冷冻鸡肉和57份农贸市场环境样品分别检出28份、2份和4份空肠弯曲菌阳性,阳性率分别为17.28%、1.12%和7.02%;418份腹泻患者粪便,9份检出空肠弯曲菌,阳性率为2.15%;857份健康体检人群的肛拭子未检出空肠弯曲菌。结论:在除健康体检人群肛拭子外的7大类样品中均检出

  5. A role for tungsten in the biology of Campylobacter jejuni: tungstate stimulates formate dehydrogenase activity and is transported via an ultra-high affinity ABC system distinct from the molybdate transporter.

    Smart, Jonathan P; Cliff, Matthew J; Kelly, David J

    2009-11-01

    The food-borne pathogen Campylobacter jejuni possesses no known tungstoenzymes, yet encodes two ABC transporters (Cj0300-0303 and Cj1538-1540) homologous to bacterial molybdate (ModABC) uptake systems and the tungstate transporter (TupABC) of Eubacterium acidaminophilum respectively. The actual substrates and physiological role of these transporters were investigated. Tryptophan fluorescence spectroscopy and isothermal titration calorimetry of the purified periplasmic binding proteins of each system revealed that while Cj0303 is unable to discriminate between molybdate and tungstate (K(D) values for both ligands of 4-8 nM), Cj1540 binds tungstate with a K(D) of 1.0 +/- 0.2 pM; 50 000-fold more tightly than molybdate. Induction-coupled plasma mass spectroscopy of single and double mutants showed that this large difference in affinity is reflected in a lower cellular tungsten content in a cj1540 (tupA) mutant compared with a cj0303c (modA) mutant. Surprisingly, formate dehydrogenase (FDH) activity was decreased approximately 50% in the tupA strain, and supplementation of the growth medium with tungstate significantly increased FDH activity in the wild type, while inhibiting known molybdoenzymes. Our data suggest that C. jejuni possesses a specific, ultra-high affinity tungstate transporter that supplies tungsten for incorporation into FDH. Furthermore, possession of two MoeA paralogues may explain the formation of both molybdopterin and tungstopterin in this bacterium. PMID:19818021

  6. The Effects of Temperature and Innate Immunity on Transmission of Campylobacter jejuni (Campylobacterales: Campylobacteraceae) Between Life Stages of Musca domestica (Diptera: Muscidae)

    Bahrndorff, Simon; Gill, C.; Lowenberger, C.; Skovgard, H.; Hald, Birthe

    2014-01-01

    The house fly (Musca domestica L.) is a well-established vector of human pathogens, including Campylobacter spp., which can cause infection of broiler chicken flocks, and through contaminated broiler meat can cause outbreaks of campylobacteriosis in humans. We investigated whether Campylobacter j...

  7. Host factors determine anti-GM1 response following oral challenge of chickens with Guillain-Barre syndrome derived Campylobacter jejuni strain GB11.

    C Wim Ang

    Full Text Available BACKGROUND: Anti-ganglioside antibodies with a pathogenic potential are present in C. jejuni-associated Guillain-Barré syndrome (GBS patients and are probably induced by molecular mimicry. Immunization studies in rabbits and mice have demonstrated that these anti-ganglioside antibodies can be induced using purified lipo-oligosaccharides (LOS from C. jejuni in a strong adjuvant. METHODOLOGY/PRINCIPAL FINDINGS: To investigate whether natural colonization of chickens with a ganglioside-mimicking C. jejuni strain induces an anti-ganglioside response, and to investigate the diversity in anti-ganglioside response between and within genetically different chicken lines, we orally challenged chickens with different C. jejuni strains. Oral challenge of chickens with a C. jejuni strain from a GBS patient, containing a LOS that mimics ganglioside GM1, induced specific IgM and IgG anti-LOS and anti-GM1 antibodies. Inoculation of chickens with the Penner HS:3 serostrain, without a GM1-like structure, induced anti-LOS but no anti-ganglioside antibodies. We observed different patterns of anti-LOS/ganglioside response between and within the five strains of chickens. CONCLUSIONS: Natural infection of chickens with C. jejuni induces anti-ganglioside antibodies. The production of antibodies is governed by both microbial and host factors.

  8. Role of fliY gene in pathogenicity-associated chemotaxis and colonization of Campylobacter jejuni%fliY基因在空肠弯曲菌致病性相关趋化和定植中的作用

    楼宏强; 葛玉梅; 张金良; 严杰; 赵金方

    2013-01-01

    Objective; To construct a knockout fliY gene mutant strain of Campylobacter jejuni for determining the role of FliY protein in flagellar movement related to bacterial motility, chemotaxis and colonization. Methods; The plasmid pBluescript-Ⅱ-SK was used to construct the suicide plasmid; according to homologous exchange principle, the suicide plasmid was utilized to generate fliY gene knockout mutant(fliY) in Campylobacter jejuni strain NCTC11168. The fliY mutant strain was identified by PCR, sequencing and Western blotting. The chemotactic and colonizing abilities of fliY mutant were determined by colony migration test and bacterial chemotactic test in vitro, and colonization test in jejunum of mice. Results; The fliY- mutant strain showed a growth curve in medium similar to that of wild-type strain. PCR, sequencing and Western blotting assay confirmed that the fliY gene in fliY- mutant was deleted. Compared to the wild-type strain,the colonies of fliY- mutant on semisolid plate were much smaller(P <0. 05) ,the chemotactic ability of fliY mutant towards sodium deoxycholate and bovine bile was significantly attenuated ( P <0. 05 ) ,and the number of fliY mutant(CFU) in jejunal tissue specimens of the infected mice was significantly decreased(P <0.05). Conclusion; The function of C. jejuni fliY gene refers to controlling flagellar movement,which is involved in bacterial chemotaxis and colonization.%目的:构建空肠弯曲菌(Campylobacter jejuni)鞭毛马达开关蛋白FliY编码基因(fliY)敲除突变株,了解FliY蛋白控制细菌鞭毛运动的作用及其与细菌趋化和定植的关系.方法:采用pBluescript-Ⅱ-SK质粒构建用于敲除空肠弯曲菌NCTC11168株fliy基因的自杀质粒.根据同源交换原理,利用自杀质粒构建空肠弯曲菌fliY基因敲除突变株(fliY-).采用PCR、PCR产物测序与Western Blot,对fliY-突变株进行鉴定.采用体外菌落迁徙试验、细菌趋化试验以及小鼠空肠定植试验,检测fliY-突

  9. Cloning, expression, purification and antigenicity analysis of AhpC in Campylobacter jejuni%空肠弯曲菌AhpC蛋白的克隆表达及抗原性分析

    曹芳芳; 孟凡亮; 乔博; 张茂俊; 张建中

    2011-01-01

    目的 构建空肠弯曲菌烷基过氧化氢还原酶(ah pC)基因的原核表达系统,克隆表达AhpC蛋白,用Western blot以及ELISA方法检测表达蛋白的抗原性,为筛选空肠弯曲菌感染后特异性抗体检测试剂的制备奠定基础.方法 用PCR方法从中国分离菌株ICDCCJ07001的染色体DNA中扩增出ah pC基因,将目的基因插入表达载体pGEX-4T-1后转化入大肠杆菌E.coliBL21( DE3),IPTG诱导重组质粒pG EX-ah pC的表达.SDS-PAGE分析表达产物,采用GST标签亲和层析柱纯化表达蛋白.应用兔免疫血清、临床感染者血清以及健康无感染者血清,利用Western blot以及ELISA的方法分析AhpC蛋白的抗原性并初步评价其在ELISA检测方法中的应用.结果 PCR扩增及双酶切鉴定证实重组质粒构建成功.重组表达蛋白经飞行质谱鉴定为空肠弯曲菌AhpC蛋白,Western blot及ELISA检测结果显示AhpC具有特异抗原性.结论 成功构建了ah pC重组表达载体pGEX-4T-1/ahpC,并在大肠杆菌中高效表达.空肠弯曲菌AhpC蛋白具有抗原性,可以作为空肠弯曲菌感染后血清抗体检测的特异抗原.%To clone and express the AhpC protein of Campylobacter jejuni in E. Coli and characterize its antigenicity and specificity. The ah pc gene was amplified by PCR from C. Jejeuni strain ICDCCJ07001 < and cloned into pGEX-4T-l. The recombinant PGEX-4T-l-ah pC was transformed to E. Coli BL21(DE3) for protein expression. The expression product was analyzed and identified by SDS-PAGE and MALD1 TOF-TOF 4700 Proteomics Analyzer. The expressed protein was identified as Campylobacter jejuni AhpC protein by Proteomics Analyzer. Both Western blot and ELISA showed that AhpC had specific antigenicity for Campylobacter jejuni. The expression and purification of recombinant AhpC antigen with good antigenicity and specificity will facilitate the research and clinical application in the diagnosis of C. Jejuni infection.

  10. 空肠弯曲杆菌中喹诺酮类抗生素耐药基因检测与耐药性研究%Study on the Quinolone Antibiotic Resistance and Its Relative Genes in Campylobacter Jejuni

    陈云鹏; 林雯; 邓建平

    2015-01-01

    目的:建立一种检测人粪便样本中,空肠弯曲杆菌含喹诺酮类抗生素耐药基因情况的实时荧光 PCR 方法,并对空肠弯曲杆菌对喹诺酮类抗生素耐药性与耐药基因的相关性进行初步研究。方法根据空肠弯曲杆菌对喹诺酮类抗生素耐药基因 gyrA 和 gyrB 序列设计引物,建立对应的实时荧光 PCR 方法,对79例空肠弯曲杆菌进行检测,基因扩增产物经琼脂糖凝胶电泳鉴定。以药敏试验结果作为参照标准,对两种检测方法的结果进行统计学分析,计算实时荧光 PCR 的主要技术指标。结果22例(27.8%)空肠弯曲杆菌的实时荧光 PCR 出现特异性扩增曲线,扩增产物电泳结果显示其中13例携带 gyrA 基因(59.1%),7例携带 gyrB 基因(31.8%),2例同时携带 gyrA 和 gyrB 基因(9.1%)。药敏试验显示26例空肠弯曲杆菌为环丙沙星耐药型(32.9%)。统计学分析显示,两种方法的检测结果差异无统计学意义(χ2=1.125,P >0.05),一致性较好。实时荧光 PCR 法的灵敏度和特异度分别为76.9%和96.2%,总符合率为89.9%。结论实时荧光PCR 法能够检测空肠弯曲杆菌耐药基因 gyrA 和 gyrB。耐药基因与耐药型之间有所关联,需要进一步研究。%Objective To establish a real-time fluorescence PCR method to detect the drug resistance genes of pathogenic Campylobacter jejunum in human stool samples,and investigate the relationship between quinoloneantibiotic resistance and the related genes in Campylobacter jejuni .Methods According to the gyrA and gyrB gene sequences that related with the fluoroquinolone resistance in Campylobacter jejuni ,the primers of the PCR method was designed and synthesized.A rapid real-time fluorescence PCR method to detect the drug resistance genes in Campylobacter jejuni samples was established,and the optimum reaction system and conditions were screened through an

  11. 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

    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

  12. Flies and Campylobacter infection of broiler flocks

    Hald, Birthe; Skovgård, Henrik; Bang, Dang Duong;

    2004-01-01

    A total of 8.2% of flies caught outside a broiler house in Denmark had the potential to transmit Campylobacter jejuni to chickens, and hundreds of flies per day passed through the ventilation system into the broiler house. Our study suggests that flies may be an important source of Campylobacter...

  13. Flies and Campylobacter infection of broiler flocks

    Hald, Birthe; Skovgård, Henrik; Bang, Dang Duong; Pedersen, Karl; Dybdahl, Jens; Jespersen, Jørgen B.; Madsen, Mogens

    2004-01-01

    A total of 8.2% of flies caught outside a broiler house in Denmark had the potential to transmit Campylobacter jejuni to chickens, and hundreds of flies per day passed through the ventilation system into the broiler house. Our study suggests that flies may be an important source of Campylobacter...... infection of broiler flocks in summer....

  14. Medium-chain fatty acids and plant-derived antimicrobials to prevent Campylobacter colonization in broiler chickens

    Hermans, David; Haesebrouck, Freddy; Van Deun, Kim; Martel, An; Verlinden, Marc; Garmyn, An; Heyndrickx, Marc; Pasmans, Frank

    2011-01-01

    Campylobacter jejuni is the most common cause of bacterial-mediated diarrhoeal disease worldwide. Because poultry products are a major source of C. jejuni infections in humans, efforts should be taken to develop strategies to decrease Campylobacter colonization of poultry during primary production. Organic acids and plant-derived antimicrobial compounds possess marked bactericidal activity toward C. jejuni in vitro and might therefore have potential as feed additives to control C. jejuni colo...

  15. 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

    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.

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

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

    2015-01-01

    During the last years, Campylobacter has emerged as the leading cause of bacterial foodborne infections in developed countries. Described as an obligate microaerophile, Campylobacter has puzzled scientists by surviving a wide range of environmental oxidative stresses on foods farm to retail, and thereafter intestinal transit and oxidative damage from macrophages to cause human infection. In this study, confocal laser scanning microscopy (CLSM) was used to explore the biofilm development of tw...

  17. [Characterization of Campylobacter spp. from wild birds].

    Glünder, G

    1989-02-01

    Bacteria of the genus Campylobacter were isolated from 28 Rooks (Corvus frugilegus), 1 Red Kite (Milvus milvus), 1 Lapwing (Vanellus vanellus), 1 Coot (Fulica atra), 1 Common Moorhen (Gallinula chloropus) and 1 Northern Mallard (Anas platyrhynchos). Altogether, C. jejuni biovar 1, was isolated 19x, C. jejuni biovar 2 8x and C. coli 5x. Among C. jejuni biovar 1 and 2 there were 5 isolates tolerating a content of 1.5% NaCl in the medium. H2S proof of 3 C. jejuni biovar 2 and 1 C. coli isolates resulted positive or negative dependent on incubation time of the used bacterial inoculum. Concerning Rooks the findings indicate that nestlings are more often infected with campylobacters than older birds. Only 1 campylobacter isolate could be recovered from altogether 54 birds of prey although 16 Buzzards (Buteo buteo) were investigated as nestlings. PMID:2930449

  18. Longitudinal study of the excretion patterns of thermophilic Campylobacter spp. in young pet dogs in Denmark

    Hald, Birthe; Pedersen, Karl; Wainø, Michael;

    2004-01-01

    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...... concurrent minor mutative changes) or independent strains. In contrast, the excreted C. jejuni isolates were much more diverse and, in most cases, only seen in one sample from each dog. A high degree of diversity among different dogs was seen. We conclude that young domestic pet dogs excreted Campylobacter...

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

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

    2011-01-01

    A chaperone activity is sufficient for growth at high temperature or oxidative stress, whereas the HtrA protease activity is only essential at conditions close to the growth limit for C. jejuni. However, the protease activity was required to prevent induction of the cytoplasmic heat-shock response even at...

  20. Field handling conditions of raw milk sold in vending machines: experimental evaluation of the behaviour of Listeria monocytogenes, Escherichia coli O157:H7, Salmonella Typhimurium and Campylobacter jejuni

    Renato G. Zanoni

    2012-01-01

    Full Text Available The direct sale by farmers of raw milk for human consumption has been allowed in Italy since 2004. The aim of this study was to evaluate the behaviour of selected foodborne pathogens in raw milk sold in vending machines, in field handling conditions, and during shelf-life from production to consumption. Temperature of storage of raw milk in 33 farms authorized to produce and sell raw milk were investigated from farm to vending machine delivery, together with consumer habits in one province of the Emilia-Romagna region of northern Italy. Failure to maintain appropriate low temperatures during shelf-life was recorded and 43% of consumers did not boil milk before consumption. Listeria monocytogenes, Escherichia coli O157:H7, Salmonella Typhimurium and Campylobacter jejuni strains were inoculated into raw milk samples, and the best (4°C as established by law and worst temperature storage conditions detected (variable temperature were simulated. Boiling tests were performed for each pathogen considered at high and low levels of contamination. Results showed an increase in L. monocytogenes in milk stored at 4°C and at variable temperatures recorded in shelf-life monitoring, an increase in E. coli O157:H7 and S. Typhimurium at variable temperatures but not at 4°C, and a decrease in C. jejuni in all storage conditions. Boiling milk is effective in making it safe for consumers. This study provides evidence that appropriate handling of raw milk, maintaining low temperatures, together with consumer education concerning boiling raw milk before consumption are key factors in preventing foodborne infections linked to raw milk consumption, and helps assess the risk of foodborne infection linked to raw milk consumption.