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Sample records for bacterium campylobacter jejuni

  1. Campylobacter jejuni organism (image)

    Science.gov (United States)

    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. Inflammasome activation by Campylobacter jejuni

    NARCIS (Netherlands)

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

    2014-01-01

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

  3. Reactions of Chicken Sera to Recombinant Campylobacter jejuni Flagellar Proteins

    Science.gov (United States)

    Campylobacter jejuni is a Gram-negative rod bacterium and is the leading but under-reported bacterial food-borne pathogen that causes human campylobacteriosis worldwide. Raw or undercooked poultry products are regarded as a major source for human infection. C. jejuni flagella have been implicated ...

  4. Campylobacter jejuni : An emerging pathogen

    Directory of Open Access Journals (Sweden)

    Nathanon Trachoo

    2003-01-01

    Full Text Available Campylobacter jejuni is a major cause of food-borne diarrhea in many countries. However, in some countries, a number of cases were undetected because of the inappropriate detection method and ignorance. Although C. jejuni usually does not cause death in health adults, it can be deadly for immunocompromised persons (Pigrau, et al., 1997. Although thought to be very susceptible in several conditions, C. jejuni in fact is quite prevalent in nature. It can easily cause sporadic cases and outbreaks resulting in economic loss. This review covers three major parts: clinical aspects of Campylobacteriosis, C. jejuni reservoirs and transmission, and methods for detection.

  5. PRESENCE OF RESISTANCE IN CAMPYLOBACTER JEJUNI AND CAMPYLOBACTER COLI

    OpenAIRE

    Branislava Kocić; Biljana Miljković-Selimović; Tatjana Babić; Ljiljana Ristić

    2009-01-01

    There are 18 species belonging to the genus of Campylobacter (rRNK group I), of which thermophilic ones are the following: Campylobacter jejuni, Campylobacter coli, Campylobacter lari and Campylobacter upsaliensis. The aim of our research was to determine the sensitivity of Campylobacter species, isolated from human feces, to antibiotics being used in practice. The study involved 50 human strains of C. jejuni/coli isolated from feces in the Center for Microbiology in the Public Health Insti...

  6. Resistance mechanisms in Campylobacter jejuni

    Science.gov (United States)

    Iovine, Nicole M.

    2013-01-01

    Campylobacter jejuni is a major cause of food-borne gastroenteritis worldwide. While mortality is low, morbidity imparted by post-infectious sequelae such as Guillain-Barré syndrome, Reiter syndrome/reactive arthritis and irritable bowel syndrome is significant. In addition, the economic cost is high due to lost productivity. Food animals, particularly poultry, are the main reservoirs of C. jejuni. The over-use of antibiotics in the human population and in animal husbandry has led to an increase in antibiotic-resistant infections, particularly with fluoroquinolones. This is problematic because C. jejuni gastroenteritis is clinically indistinguishable from that caused by other bacterial pathogens, and such illnesses are usually treated empirically with fluoroquinolones. Since C. jejuni is naturally transformable, acquisition of additional genes imparting antibiotic resistance is likely. Therefore, an understanding of the antibiotic resistance mechanisms in C. jejuni is needed to provide proper therapy both to the veterinary and human populations. PMID:23406779

  7. The influence of environmental conditions on the infectivity of non-growing Campylobacter jejuni

    NARCIS (Netherlands)

    Verhoeff-Bakkenes L; Leusden FM van; Jonge R de; MGB

    2004-01-01

    Campylobacter (C.) jejuni is a causative agent of gastro-enteritis. Growth of this Gram negative bacterium is restricted to a limited number of environmental conditions. Under conditions where growth of C. jejuni is not possible, e.g. at temperatures below 30 degrees C, C. jejuni can remain viable

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

    DEFF Research Database (Denmark)

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

    2015-01-01

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

  9. Campylobacter jejuni in commercial eggs.

    Science.gov (United States)

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

    2014-01-01

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

  10. Campylobacter jejuni in commercial eggs

    Directory of Open Access Journals (Sweden)

    Belchiolina Beatriz Fonseca

    2014-01-01

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

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

    OpenAIRE

    Noormohamed, Aneesa; Fakhr, Mohamed

    2013-01-01

    Studies that investigate arsenic resistance in the foodborne bacterium Campylobacter are limited. A total of 552 Campylobacter isolates (281 Campylobacter jejuni and 271 Campylobacter coli) isolated from retail meat samples were subjected to arsenic resistance profiling using the following arsenic compounds: arsanilic acid (4?2,048 ?g/mL), roxarsone (4?2048 ?g/mL), arsenate (16?8,192 ?g/mL) and arsenite (4?2,048 ?g/mL). A total of 223 of these isolates (114 Campylobacter jejuni and 109 Campyl...

  12. Distribution of Flagella Secreted Protein and Integral Membrane Protein Among Campylobacter jejuni Isolated from Thailand

    Science.gov (United States)

    2011-01-01

    secreted protein and integral membrane protein among Campylobacter jejuni isolated from Thailand Piyarat Pootong 1·, Oralak Serichantalergs...Ladaporn Bodhidatta \\ Frederic Poly2, Patricia Guerry2 and Carl J Mason 1 Abstract Background: Campylobacter jejuni, a gram-negative bacterium, is a...groups of integral membrane protein. The significance of these different FspA variants to virulence requires further study. Background Campylobacter

  13. Studies on the epidemiology of Campylobacter jejuni

    NARCIS (Netherlands)

    J. Oosterom (Johannes)

    1985-01-01

    textabstractOver the last few years the bacterial species Campylobacter jejuni has been recognized as an important cause of acute enteritis in man. Investigations in several countries have shown that infections caused by C. jejuni may be as serious as those due to Salmonella spp., both in prevalence

  14. Campylobacter jejuni strategies to evade hostile environments

    NARCIS (Netherlands)

    Vaezirad, M.M.

    2017-01-01

    Campylobacter jejuni is the most common cause of human bacterial foodborne disease in the western world. Each year hundreds of millions of cases of Campylobacter infection occur worldwide. After a few weeks, the infection may be followed by serious auto-immune diseases like the Guillain-Barre

  15. Antimicrobial resistance profiles of Campylobacter jejuni isolates from wild birds in Sweden

    NARCIS (Netherlands)

    Waldenstrom, J.; Mevius, D.J.; Veldman, K.T.; Broman, T.; Hasselquist, D.; Olsen, B.

    2005-01-01

    In order to determine the occurrence and frequency of resistant strains of the bacterium Campylobacter jejuni and to establish baseline MICs ta isolates from an environmental reservoir, the resistance profiles of 10 antimicrobial substances were determined for 137 C. jejuni isolates from wild birds

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

    NARCIS (Netherlands)

    A.P. Heikema (Astrid)

    2013-01-01

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

  17. Genetic Characterization of Campylobacter Jejuni and C. coli Isolated From Broilers Using flaA PCR-Restriction Fragment Length Polymorphism Method in Shiraz, Southern Iran

    OpenAIRE

    Khoshbakht, Rahem; Tabatabaei, Mohammad; Hosseinzadeh, Saeid; Shirzad Aski, Hesamaddin; Seifi, Saeed

    2015-01-01

    Background: Thermophilic campylobacters, particularly Campylobacter jejuni and C. coli are the main agents of human campylobacteriosis. Campylobacter contaminated chicken products is the most important source of foodborne gastroenteritis. Evaluation of genetic diversity among Campylobacter population is critical for understanding the epidemiology of this bacterium and developing effective control strategies against Campylobacter infections and other related disorders. Objectives: The aim of t...

  18. Campylobacter jejuni - A MONOGRAPHIC STUDY (REVIEW

    Directory of Open Access Journals (Sweden)

    N. CORCIONIVOSCHI

    2009-05-01

    Full Text Available Campylobacter is the primary cause of bacterial diarrhoeal illness in the developed world with an estimated 2-3 million Campylobacter-related illnesses occurring in the United States per year. Campylobacter jejuni can cause a spectrum of disease including gastroenteritis, proctitis, septicaemia, meningitis, abortion and autoimmune diseases such as Reiter’s arthritis and Guillain-Barré syndrome (GBS. The association of Campylobacter with poultry (e.g. chickens, turkeys, ducks, and geese has been known for the last 30 years. In this review we will present the biology of this organism as presented for the last two decades and also the connection between Campylobacter jejuni and farm animals.

  19. Surface Plasmon Resonance Immunosensor for the Detection of Campylobacter jejuni

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    Noor Azlina Masdor

    2017-05-01

    Full Text Available Campylobacteriosis is an internationally important foodborne disease caused by Campylobacter jejuni. The bacterium is prevalent in chicken meat and it is estimated that as much as 90% of chicken meat on the market may be contaminated with the bacterium. The current gold standard for the detection of C. jejuni is the culturing method, which takes at least 48 h to confirm the presence of the bacterium. Hence, the aim of this work was to investigate the development of a Surface Plasmon Resonance (SPR sensor platform for C. jejuni detection. Bacterial strains were cultivated in-house and used in the development of the sensor. SPR sensor chips were first functionalized with polyclonal antibodies raised against C. jejuni using covalent attachment. The gold chips were then applied for the direct detection of C. jejuni. The assay conditions were then optimized and the sensor used for C. jejuni detection, achieving a detection limit of 8 × 106 CFU·mL−1. The sensitivity of the assay was further enhanced to 4 × 104 CFU·mL−1 through the deployment of a sandwich assay format using the same polyclonal antibody. The LOD obtained in the sandwich assay was higher than that achieved using commercial enzyme-linked immunosorbent assay (ELISA (106–107 CFU·mL−1. This indicate that the SPR-based sandwich sensor method has an excellent potential to replace ELISA tests for C. jejuni detection. Specificity studies performed with Gram-positive and Gram-negative bacteria, demonstrated the high specific of the sensor for C. jejuni.

  20. PRESENCE OF RESISTANCE IN CAMPYLOBACTER JEJUNI AND CAMPYLOBACTER COLI

    Directory of Open Access Journals (Sweden)

    Branislava Kocić

    2009-04-01

    Full Text Available There are 18 species belonging to the genus of Campylobacter (rRNK group I, of which thermophilic ones are the following: Campylobacter jejuni, Campylobacter coli, Campylobacter lari and Campylobacter upsaliensis. The aim of our research was to determine the sensitivity of Campylobacter species, isolated from human feces, to antibiotics being used in practice. The study involved 50 human strains of C. jejuni/coli isolated from feces in the Center for Microbiology in the Public Health Institute Nis. Sensitivity was tested by applying the disk diffusion method on seven antibiotics (erythromycin, gentamicin, tetracycline, ciprofloxacin, hloramphenicol, cephalexin and nalidixic acid. Our results showed low resistance to erythromycin, gentamicin and tetracycline (2%, which corresponds to the studies conducted in the world. Moreover, these findings indicate that erythromycin may be considered the drug of choice in the treatment of Campylobacter diarrhea in this region. Resistance to fluoroquinolone and nalidixic acid was 44%, and C. coli showed higher resistance compared to C. jejuni, though statistical significance was not proved.

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

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

    2016-01-01

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

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

    Science.gov (United States)

    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. Copyright © 2015 Elsevier GmbH. All rights reserved.

  3. Campylobacter jejuni seroepidemiology in native chicken

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    Anwar Rosyidi

    2012-10-01

    Full Text Available Campylobacter jejuni is responsible for about 90% of cases of Campylobacteriosis in humans with gastroenteritis. Healthy chickens can carry Campylobacter spp. in the intestinal tract. Efforts to reduce exposure to Campylobacteriosis by humans may be enhanced by knowledge of its prevalence in poultry. This study aimed to identify factors associated with seropositive response to C. jejuni in native chickens in Mataram. Detection of C. jejuni was accomplished using an immunochromatographic serological method. Association between Campylobacter jejuni seropositive response as the dependent variable with various independent variables was analyzed using χ² (Chi square and Odds Ratio (OR. A total of 216 chicken samples were examined and 44 chicken owners were interviewed and their farms examined. Results showed the prevalence of serological response to C. jejuni in chicken samples to be as high as 35.6% and that as many as 70.5% of farms had affected chickens. Age of the chicken was the variable most closely associated with incidence of seropositive response, birds older than 3 months more likely to be affected. Variables at the farm level associated with variation in seropositive response were cage type, cage floor material, and origin of drinking water, surface water sources being less desirable.

  4. Campylobacter jejuni diarrhea model in infant chickens

    NARCIS (Netherlands)

    Sanyal, S. C.; Islam, K. M.; Neogy, P. K.; Islam, M.; Speelman, P.; Huq, M. I.

    1984-01-01

    To study the pathogenic mechanisms of Campylobacter jejuni infection, 36- to 72-h-old chickens were fed 10(3) to 10(6) live cells, using strains isolated from 40 patients with watery diarrhea and 6 with bloody mucoid diarrhea from whom no other known enteropathogen was detected. Chickens of Starbro

  5. Genomic Characterization of Campylobacter jejuni strain M1

    DEFF Research Database (Denmark)

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

    2010-01-01

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

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

    DEFF Research Database (Denmark)

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

    2012-01-01

    Campylobacter jejuni (C. jejuni) is one of the leading causes of bacterial food-borne disease worldwide. The presence of Campylobacter in chicken feces poses a high risk for contamination of chicken meat and for Campylobacter infections in human. Detection of this bacterium in chicken fecal...... 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 selection...

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

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

  8. Specific detection of Campylobacter jejuni and Campylobacter coli by using polymerase chain reaction.

    OpenAIRE

    Oyofo, B A; Thornton, S A; Burr, D H; Trust, T J; Pavlovskis, O R; Guerry, P

    1992-01-01

    Development of a routine detection assay for Campylobacter jejuni and Campylobacter coli in clinical specimens was undertaken by using the polymerase chain reaction (PCR). An oligonucleotide primer pair from a conserved 5' region of the flaA gene of C. coli VC167 was used to amplify a 450-bp region by PCR. The primer pair specifically detected 4 strains of C. coli and 47 strains of C. jejuni; but it did not detect strains of Campylobacter fetus, Campylobacter lari, Campylobacter upsaliensis, ...

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

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    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.

  10. Campylobacter jejuni : A brief overview on pathogenicity-associated factors and disease-mediating mechanisms

    NARCIS (Netherlands)

    Dasti, Javid I.; Tareen, A. Malik; Lugert, Raimond; Zautner, Andreas E.; Gross, Uwe

    Campylobacter jejuni has long been recognized as a cause of bacterial food-borne illness, and surprisingly, it remains the most prevalent bacterial food-borne pathogen in the industrial world to date. Natural reservoirs for this Gram-negative, spiral-shaped bacterium are wild birds, whose intestines

  11. Persistence of Two Campylobacter jejuni Strains in Soil and on Spinach Plants

    International Nuclear Information System (INIS)

    Jaderlund, L.; Arthurson, V.; Sessitsch, A.

    2011-01-01

    There are indications that the more frequent use of untreated organic residues for fertilization results in increased risk of contamination with human pathogens. Here, we evaluate the ability of two different strains of Campylobacter jejuni to persist in manure and soil as well as spread to spinach plants. It was revealed that different strategies for inoculation of C. jejuni contribute to the persistence of the bacterium in soil, roots, and shoots. Upon inoculation of the bacteria into manure prior to soil application, the amount of C. jejuni subsequently recovered in soil was higher than that from treatments involving the addition of C. jejuni cells to the soil after plant emergence. Irrespective of the bacterial inoculation dose and strategy employed, the C. jejuni content in soil remained relatively constant, whereas the majority of C. jejuni cells applied to spinach leaves could be recovered during the whole evaluation period of 21 days.

  12. Molecular Mechanisms Influencing Genetic Diversity of Campylobacter jejuni

    NARCIS (Netherlands)

    Gaasbeek, E.J.

    2009-01-01

    Campylobacter jejuni is an important food-borne pathogen, causing human bacterial gastroenteritis. Throughout the years several methods have been developed for typing C. jejuni. These methods uncovered the existence of enormous genetic diversity within the species. Stable lineages of C. jejuni are

  13. Clinical aspects of Campylobacter jejuni infections in adults.

    OpenAIRE

    Peterson, M C

    1994-01-01

    Campylobacter jejuni is an almost ubiquitous, microaerophilic, gram-negative rod. Outbreaks have been associated with drinking raw milk or contaminated water and eating poultry. Campylobacter jejuni accounts for 3.2% to 6.1% of cases of diarrheal illness in the general population of the United States, and infected patients frequently present with abdominal pain and fever. Less frequently, C jejuni is responsible for bacteremia, septic arthritis, septic abortion, and other extraintestinal infe...

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

    NARCIS (Netherlands)

    Verhoeff-Bakkenes, L.

    2012-01-01

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

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

    NARCIS (Netherlands)

    Verhoeff-Bakkenes, L.

    2012-01-01

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

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

    Science.gov (United States)

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

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

  17. Effect of gamma radiation on Campylobacter jejuni

    International Nuclear Information System (INIS)

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

    1984-01-01

    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 0 C, at 0-5 0 C, and at 30 +/- 10 0 C. Irradiation at -30 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 10 value for C. jejuni was 32 Krad, which was less than D 10 values commonly reported for salmonellae. 20 references, 4 figures

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

    NARCIS (Netherlands)

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

    2003-01-01

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

  19. Campylobacter jejuni infection in broiler chickens.

    Science.gov (United States)

    Dhillon, A Singh; Shivaprasad, H L; Schaberg, D; Wier, F; Weber, S; Bandli, D

    2006-03-01

    Day-old, straight-run broiler chickens were procured from a hatchery located in the Pacific Northwest. The chickens were subdivided individually into nine groups of 20 chickens. The chickens were tagged, housed in isolation chambers on wire, fed commercial broiler feed, and given water ad libitum. Three isolates of Campylobacter jejuni of poultry origin and one of human origin were tested in this study. Various C. jejuni cultures were inoculated into 9-day-old chickens by crop gavage. Four groups of 20 chickens were inoculated at a dose level of 0.5 ml of 1 x 10(2) colony-forming units (CFU)/ml. The other four groups were inoculated with 0.5 ml of 1 X 10(4) CFU/ml. One group of 20 chickens was kept as an uninoculated control group. Four randomly selected chickens from each of the inoculated and uninoculated groups were necropsied at 5, 12, and 19 days postinoculation (DPI). The C. jejuni was cultured and enumerated from a composite of the upper and midintestine and the cecum. Body weights of all chicken groups at 7 days of age and at 5, 12, and 19 DPI were measured and statistically analyzed. No significant differences were present in the mean body weights (MBWs) of 7-day-old, 5 DPI, and 12 DPI male and female broiler chickens inoculated with C. jejuni at both dose levels compared with uninoculated controls. Differences in MBWs of the male and female broilers at 19 DPI were observed in some of the groups. Results of the C. jejuni culture enumeration mean (CEM) of composite intestine samples at 5 DPI from all inoculated chicken groups, irrespective of the dose level, ranged from (2.5 +/- 5.0) x 10(2) to (2.8 +/- 4.8) x 10(5) CFU/g (mean +/- SD). Results of cecum C. jejuni CEM at 5 DPI inoculated at both dose levels ranged from (2.5 +/- 5.0) x 10(6) to (1 +/- 0.0) x 10(7) CFU/g in all treatment groups irrespective of the dose level. CEM results from the composite intestine samples at 12 and 19 DPI increased by 1 log unit, or sometimes more. Results of cecum C. jejuni

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

    DEFF Research Database (Denmark)

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

    2003-01-01

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

  1. Methods for Initial Characterization of Campylobacter jejuni Bacteriophages.

    Science.gov (United States)

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

    2017-01-01

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

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

    Directory of Open Access Journals (Sweden)

    Mohamed K. Fakhr

    2013-08-01

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

  3. Methods for initial characterization of Campylobacter jejuni bacteriophages

    DEFF Research Database (Denmark)

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

    2017-01-01

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

  4. Cytolethal Distending Toxin Genes in Campylobacter jejuni and Campylobacter coli Isolates: Detection and Analysis by PCR

    OpenAIRE

    Eyigor, Aysegul; Dawson, Karl A.; Langlois, Bruce E.; Pickett, Carol L.

    1999-01-01

    Campylobacter jejuni produces a toxin called cytolethal distending toxin (CDT). Knowledge of the prevalence and homogeneity of Campylobacter sp. cdt genes is incomplete. In this work, we identified four PCR primer pairs that collectively amplified cdt genes in all of the C. jejuni and Campylobacter coli strains tested. Restriction analyses of the cdt PCR products showed clear differences between the cdt genes of these two species, yet there were few heterogeneities noted between members of th...

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

    Directory of Open Access Journals (Sweden)

    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.

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

    DEFF Research Database (Denmark)

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

    2009-01-01

    This study evaluated the effectiveness of 11 chemical compounds to reduce Campylobacter jejuni on chicken skin and meat samples dipped in chemical solutions. Treatment of skin samples for 1 min using tartaric acid (2%) and caprylic acid sodium salt (5%) caused reductions of C. jejuni NCTC11168...

  7. Phage therapy reduces Campylobacter jejuni colonization in broilers

    NARCIS (Netherlands)

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

    2005-01-01

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

  8. Clonal Distribution and Virulence of Campylobacter jejuni Isolates in Blood

    Science.gov (United States)

    de Haan, Caroline P.A.; Ellström, Patrik; Sarna, Seppo; Hänninen, Marja-Liisa; Rautelin, Hilpi

    2013-01-01

    Campylobacter jejuni bacteria are highly diverse enteropathogens. Seventy-three C. jejuni isolates from blood collected in Finland were analyzed by multilocus sequence typing and serum resistance. Approximately half of the isolates belonged to the otherwise uncommon sequence type 677 clonal complex. Isolates of this clonal complex were more resistant than other isolates to human serum. PMID:24047729

  9. Survival of Campylobacter jejuni in different gas mixtures

    DEFF Research Database (Denmark)

    Boysen, Louise; Knochel, Susanne; Rosenquist, Hanne

    2007-01-01

    Campylobacter jejuni in fresh chilled chicken meat is known to be a major risk factor for human gastrointestinal disease. In the present study, the survival under chilled conditions of different C. jejuni strains exposed to different gas mixtures usually used for gas packaging of food was examined...

  10. Cytotoxity of cell free filtrates of campylobacter jejuni isolated in ...

    African Journals Online (AJOL)

    Culture filtrates of Campylobacter jejuni strains isolated from clinical specimens in Lagos Nigeria were tested for toxic activity. Two out of five filtrates tested manifested cytopathic effect on BHK cells. The effects were mainly cytotoxic and cytotonic. Toxic activity of C. jejuni filtrates was much lower than toxic activity elicited by ...

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

    OpenAIRE

    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...

  12. Methods to Study Antimicrobial Resistance in Campylobacter jejuni.

    Science.gov (United States)

    Sahin, Orhan; Shen, Zhangqi; Zhang, Qijing

    2017-01-01

    Campylobacter jejuni is a leading bacterial cause of foodborne gastroenteritis worldwide and is increasingly resistant to clinically important antibiotics. Detection of antibiotic resistance in C. jejuni can be performed with both phenotypic and genotypic methods. In this chapter, we describe the most commonly used molecular biology methods for detection of resistance to clinically important antibiotics. These methods can be employed in both clinical and research settings to facilitate clinical therapy and to monitor the emergence and dissemination of antibiotic-resistant C. jejuni.

  13. Aplicación de PCR-RFLP para subtipificar Campylobacter jejuni PCR-RFLP for Campylobacter jejuni subtyping

    Directory of Open Access Journals (Sweden)

    G. Giacoboni

    2005-06-01

    Full Text Available Diez cepas de Campylobacter jejuni aisladas de fetos porcinos abortados fueron identificadas por pruebas bioquímicas: 8 como C. jejuni biotipo II de Lior, y 2 como C. jejuni biotipo I. Para poder subtipificarlas se utilizó la técnica de reacción en cadena de la polimerasa (PCR para amplificar el gen flaA y al producto obtenido se lo digirió con la enzima de restricción DdeI (RFLP. Se pudieron obtener 6 subtipos a partir de C. jejuni biotipo II, mientras que los dos aislamientos de biotipo I correspondieron a un mismo subtipo. Aunque existe una amplia variedad de técnicas de biología molecular que son aplicadas con fines epidemiológicos para Campylobacter, PCR-RFLP, demostró ser una técnica simple y accesible, capaz de subtipificar a C. jejuni.Ten Campylobacter jejuni isolates, 8 identified as C. jejuni biotype II of Lior and 2 as C. jejuni biotipe I, were recovered from aborted pig fetuses. In order to discriminate among strains, restriction fragment length polymorphism (RFLP using DdeI of polymerase chain reaction (PCR products of flaA gen was used. C. jejuni biotype II strains could be diferenciated in 6 by PCR-RFLP, and one subtype was obtained from C. jejuni biotype I. Although there is great variability of molecular techniques applied to the Campylobacter epidemiological studies, PCR-RFLP demonstrated to be a simple and accessible technique to discriminate Campylobacter jejuni isolates.

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

    OpenAIRE

    Taylor, D E; Chang, N

    1991-01-01

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

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

    DEFF Research Database (Denmark)

    Mortensen, Ninell P; Schiellerup, Peter; Boisen, Nadia

    2011-01-01

    The role of Campylobacter jejuni cytolethal distending toxin (CDT) on clinical outcome after gastroenteritis was investigated. Clinical data, blood serum samples, and Campylobacter spp. isolated, from each of 30 patients were collected over a period of 6 months. The CDT encoding genes, cdt...

  16. Cellular response of Campylobacter jejuni to trisodium phosphate

    DEFF Research Database (Denmark)

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

    2012-01-01

    The highly alkaline compound trisodium phosphate (TSP) is used as an intervention to reduce the load of Campylobacter on poultry meat in U.S. poultry slaughter plants. The aim of the present study was to investigate the cellular responses of Campylobacter jejuni NCTC11168 when exposed to sublethal...

  17. Generation of Campylobacter jejuni genetic diversity in vivo

    NARCIS (Netherlands)

    Boer, de P.; Wagenaar, J.A.; Achterberg, R.P.; Putten, van J.P.M.; Schouls, L.M.; Duim, B.

    2002-01-01

    Molecular epidemiology studies suggest that horizontal genetic exchange is a major cause of pathogen biodiversity. We tested this concept for the bacterial enteropathogen Campylobacter jejuni by seeking direct in vivo evidence for the exchange of genetic material among Campylobacter strains. For

  18. Survey of chicken abattoir for the presence of Campylobacter jejuni and Campylobacter coli Pesquisa de Campylobacter jejuni e Campylobacter coli em abatedouros de aves

    Directory of Open Access Journals (Sweden)

    Ana L.L. Cortez

    2006-12-01

    Full Text Available The genus Campylobacter is of great importance to public health because it includes several species that may cause diarrhea. These species may be found in water, food and in the intestinal tract of chickens. This study investigated the presence of Campylobacter jejuni and Campylobacter coli in chicken abattoirs in São Paulo State, Brazil. A total of 288 samples of feces, feathers, scald water, evisceration water, chiller water, and the rinse water of eviscerated, not eviscerated and chilled carcasses were collected in six chicken abattoirs. Polymerase Chain Reaction (PCR was performed in Campylobacter spp.-positive isolates using the gene HIP, specific for hippuricase enzyme from Campylobacter jejuni and aspartokinase gene, specific to detect Campylobacter coli. The percentage of positive isolates of Campylobacter jejuni was 4.9% (14/288. Isolation was greater in feces samples (22%, 8/36. One sample was positive for the species C. coli. In conclusion, the results indicate that it is necessary to improve quality control for Campylobacter spp. in chicken abattoirs.O gênero Campylobacter tem grande destaque em saúde pública, principalmente por pertencerem a este gênero várias espécies que podem causar diarréia. Estas espécies podem ser encontradas em amostras de água, alimentos e no trato intestinal das aves. Este estudo investigou a presença de Campylobacter jejuni e Campylobacter coli em abatedouros de aves no Estado de São Paulo. As 288 amostras foram coletadas em seis estabelecimentos e incluíram: fezes; penas; água de escaldamento, de evisceração e de resfriamento; e água de enxaguadura de carcaça não eviscerada, eviscerada e resfriada. Após o isolamento microbiológico das amostras positivas de Campylobacter spp. foi realizada uma Reação em Cadeia da Polimerase (PCR utilizando o gene HIP, da hipuricase, específico para Campylobacter jejuni e o gene da enzima aspartoquinase, específico para Campylobacter coli. A

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

    Science.gov (United States)

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

  20. Stress response and pathogenic potential of Campylobacter jejuni cells exposed to starvation.

    Science.gov (United States)

    Klancnik, Anja; Guzej, Bernarda; Jamnik, Polona; Vucković, Darinka; Abram, Maja; Mozina, Sonja Smole

    2009-06-01

    Campylobacter jejuni is a Gram-negative, fragile, spiral bacterium, known worldwide to be a major cause of acute human enteritis. Like many other food-borne bacteria, campylobacters must be able to survive under diverse conditions both inside the host and in the environment. Understanding stress response mechanisms provides information necessary for improving food processing and strategies that enhance food safety as well as clarifying the pathogenesis of campylobacteriosis. We investigated the relation between stress response to starvation and pathogenic potential in C. jejuni. Starvation changed the morphology and physiology of C. jejuni cells. However, the lower metabolic activity of 5-h-starved culture was not a dormant state, but probably a viable but non-culturable (VBNC) form of the cells, since starved C. jejuni induced heat stress resistance. The health hazard potential of starved cells is still unclear. We showed that, in spite of starvation, C. jejuni survived in vitro within Caco-2 enterocites up to 4 days and caused systemic campylobacteriosis in vivo in a mouse model. However, bacterial numbers in investigated organs were significantly lower and the infection was resolved sooner. Our results show that nutrient insufficiency is responsible for C. jejuni transformation, influencing but not abolishing its survival and virulence properties while in the VBNC state.

  1. Isolation and characterization of Campylobacter jejuni and Campylobacter coli from domestic and wild mammals in Norway.

    Science.gov (United States)

    Rosef, O; Gondrosen, B; Kapperud, G; Underdal, B

    1983-10-01

    A total of 1,262 domestic and wild mammals from Norway were surveyed for fecal carriage of Campylobacter jejuni and Campylobacter coli. Of the five species of domestic mammals examined, the highest isolation rate was recorded among swine (100.0%), followed by sheep (8.1%) and cows (0.8%). No strains were recovered from horses or goats. Among wild mammals, C. jejuni was isolated from 1 of 23 hares, and no isolated were obtained from three species of cervids and three species of rodents. Of the 133 Campylobacter strains isolated, 114 were classified as C. coli, 18 were C. jejuni biotype 1, and 1 belonged to C. jejuni biotype 2. All 114 strains from swine were C. coli. Milk samples from 113 domestic animals with clinically diagnosed mastitis (106 cows, 5 sheep, 1 horse, and 1 pig) were negative for campylobacters.

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

    Directory of Open Access Journals (Sweden)

    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

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

    DEFF Research Database (Denmark)

    Hald, Birthe; Skovgård, Henrik; Pedersen, Karl

    2008-01-01

    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...... 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...

  4. Reduced spread of Campylobacter jejuni in broiler chickens by stimulating the bird's natural barriers.

    Science.gov (United States)

    Moen, B; Rudi, K; Svihus, B; Skånseng, B

    2012-11-01

    We have tested the effect of feed structure and feeding regime to prevent the spread of the zoonotic pathogen Campylobacter jejuni in broiler chicken flocks. Birds were offered two types of feed, control diet and a diet supplemented with 15% oat/barley hulls for structure. In addition, the birds were either fed ad libitum or intermittent. One bird in each treatment group was infected with a three-strain-mix of Camp. jejuni, and the spread of Camp. jejuni within the group was investigated. Feed structure increased the gizzard weight, delayed the spread of Camp. jejuni within the group and reduced the relative amount of Camp. jejuni in the caecum compared with the control diet. Our results show that stimulating the bird's natural barriers is a novel and promising intervention strategy to reduce the spread of Camp. jejuni in chicken flocks. Preventing Camp. jejuni in broiler chicken flocks is essential to ensure food safety because this bacterium is transferred to chicken carcasses during the slaughter process and readily survive in unprocessed poultry products. We have evaluated a novel approach for stimulation of the bird's natural barriers in the upper digestive tract with promising results. © 2012 The Authors Journal of Applied Microbiology © 2012 The Society for Applied Microbiology.

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

    Directory of Open Access Journals (Sweden)

    Jong-Chul eKim

    2015-07-01

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

  6. Quantitative Proteomics of Intracellular Campylobacter jejuni Reveals Metabolic Reprogramming

    OpenAIRE

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

    2012-01-01

    Author Summary Campylobacter jejuni is one of the most common causes of food-borne illness in the United States and a major cause of diarrheal diseases in developing countries. This pathogen can invade intestinal epithelial cells, which is very important for its ability to cause disease. Once it gains access to epithelial cells, C. jejuni becomes unable to grow under standard growth conditions, although it can grow if pre-incubated under oxygen limiting conditions. This study compares the pro...

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

    Science.gov (United States)

    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. Copyright © 2013 Elsevier B.V. All rights reserved.

  8. Metronidazole resistance in Campylobacter jejuni from poultry meat

    DEFF Research Database (Denmark)

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

    2006-01-01

    The occurrence of metronidazole resistance was investigated among Campylobacter jejuni in raw poultry meat collected from supermarkets. MICs were determined by the agar dilution procedure in the testing range of 3 to 60 mu g/ml metronidazole. The MICs showed a bimodal distribution...... with a 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 resistance....

  9. Metronidazole resistance in Campylobacter jejuni from poultry meat

    DEFF Research Database (Denmark)

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

    2006-01-01

    The occurrence of metronidazole resistance was investigated among Campylobacter jejuni in raw poultry meat collected from supermarkets. MICs were determined by the agar dilution procedure in the testing range of 3 to 60 mu g/ml metronidazole. The MICs showed a bimodal distribution with a signific......The occurrence of metronidazole resistance was investigated among Campylobacter jejuni in raw poultry meat collected from supermarkets. MICs were determined by the agar dilution procedure in the testing range of 3 to 60 mu g/ml metronidazole. The MICs showed a bimodal distribution...

  10. Prevalence of Campylobacter jejuni in poultry breeder flocks

    Directory of Open Access Journals (Sweden)

    Ludovico Dipineto

    2010-01-01

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

  11. Inaccuracy of routine susceptibility tests for detection of erythromycin resistance of Campylobacter jejuni and Campylobacter coli

    NARCIS (Netherlands)

    Beek, M.T.; Claas, E.C.J.; Mevius, D.J.; Pelt, van W.; Wagenaar, J.A.; Kuijper, E.J.

    2010-01-01

    In The Netherlands, both an increase in and regional differences in erythromycin resistance of Campylobacter jejuni and Campylobacter coli have been reported. To determine the accuracy of routine tests for erythromycin resistance, 48 erythromycin-resistant isolates from various laboratories that

  12. Temperature-dependent phenotypic variation of Campylobacter jejuni lipooligosaccharides

    LENUS (Irish Health Repository)

    Semchenko, Evgeny A

    2010-11-30

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

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

    DEFF Research Database (Denmark)

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

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

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

    Science.gov (United States)

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

    2012-01-01

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

  15. β-Resorcylic Acid, a Phytophenolic Compound, Reduces Campylobacter jejuni in Postharvest Poultry.

    Science.gov (United States)

    Wagle, B R; Arsi, K; Upadhyay, A; Shrestha, S; Venkitanarayanan, K; Donoghue, A M; Donoghue, D J

    2017-08-01

    Human Campylobacter infections, a leading foodborne illness globally, has been linked with the high prevalence of this bacterium on raw retail chicken products. Reduction of Campylobacter counts on poultry products would greatly reduce the risk of subsequent infections in humans. To this end, this study investigated the potential of the phytophenolic compound β-resorcylic acid (BR) to reduce Campylobacter counts on postharvest poultry (chicken skin or meat). Four trials in total, two each on thigh skin or breast meat, were conducted in which chicken skin or meat samples (2 ± 0.1 g; 10 samples per treatment) were inoculated with 50 μL (∼10 6 CFU per sample) of a cocktail of four wild strains of C. jejuni. After 30 min of attachment, inoculated samples were dipped in a 0, 0.5, 1, or 2% BR solution for 30 s immediately followed by vigorously vortexing the samples in Butterfield's phosphate diluent and plating the supernatant for Campylobacter enumeration. In addition, the effect of BR on the color of skin and meat samples was studied. Moreover, the change in the expression of survival and virulence genes of C. jejuni exposed to BR was evaluated. Data were analyzed by the PROC MIXED procedure of SAS (P meat samples by 1 to 3 log CFU/g compared with non-BR-treated washed controls. No significant difference in the lightness, redness, and yellowness of skin and meat samples was observed on exposure to BR wash (P > 0.05). Real-time PCR results revealed that BR treatment down-regulated expression of select genes coding for motility (motA, motB) and attachment (cadF, ciaB) in the majority of C. jejuni strains. Stress response genes (sodB, katA) were upregulated in C. jejuni S-8 (P poultry processing for reducing Campylobacter on chicken carcasses.

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

    Directory of Open Access Journals (Sweden)

    Petra Griekspoor

    2015-12-01

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

  17. Campylobacter jejuni: a brief overview on pathogenicity-associated factors and disease-mediating mechanisms.

    Science.gov (United States)

    Dasti, Javid I; Tareen, A Malik; Lugert, Raimond; Zautner, Andreas E; Gross, Uwe

    2010-04-01

    Campylobacter jejuni has long been recognized as a cause of bacterial food-borne illness, and surprisingly, it remains the most prevalent bacterial food-borne pathogen in the industrial world to date. Natural reservoirs for this Gram-negative, spiral-shaped bacterium are wild birds, whose intestines offer a suitable biological niche for the survival and dissemination of C. jejuni Chickens become colonized shortly after birth and are the most important source for human infection. In the last decade, effective intervention strategies to limit infections caused by this elusive pathogen were hindered mainly because of a paucity in understanding the virulence mechanisms of C. jejuni and in part, unavailability of an adequate animal model for the disease. However, recent developments in deciphering molecular mechanisms of virulence of C. jejuni made it clear that C. jejuni is a unique pathogen, being able to execute N-linked glycosylation of more than 30 proteins related to colonization, adherence, and invasion. Moreover, the flagellum is not only depicted to facilitate motility but as well secretion of Campylobacter invasive antigens (Cia). The only toxin of C. jejuni, the so-called cytolethal distending toxin (CdtA,B,C), seems to be important for cell cycle control and induction of host cell apoptosis and has been recognized as a major pathogenicity-associated factor. In contrast to other diarrhoea-causing bacteria, no other classical virulence factors have yet been identified in C. jejuni. Instead, host factors seem to play a major role for pathogenesis of campylobacteriosis of man. Indeed, several lines of evidence suggest exploitation of different adaptation strategies by this pathogen depending on its requirement, whether to establish itself in the natural avian reservoir or during the course of human infection. (c) 2009 Elsevier GmbH. All rights reserved.

  18. Campylobacter jejuni in Duck Faeces around Drinking Water ...

    African Journals Online (AJOL)

    Faeces from 192 ducks feeding around 10 wells and 6 ponds in peri-urban areas of Makurdi town, North-Central Nigeria were randomly sampled during the dry season period of October, 2006 to March, 2007. The samples were cultured for Campylobacter jejuni, followed by characterisation of positive samples. The overall ...

  19. Prevalence of Campylobacter jejuni in duck faeces around drinking ...

    African Journals Online (AJOL)

    Faeces from one hundred and ninety-two ducks feeding around ten wells and six ponds in peri urban areas of Makurdi town, North-Central Nigeria were randomly sampled during the dry season period of October, 2004 to March, 2005. In total, one hundred and ninety-two samples were cultured for Campylobacter jejuni, ...

  20. The aroA gene of Campylobacter jejuni

    NARCIS (Netherlands)

    M.M.S.M. Wösten (M. M S M); H.J. Dubbink (Erik Jan); B.A.M. van der Zeijst (Ben)

    1996-01-01

    textabstractThe gene for 5-enolpyruvylshikimate-3-phosphate (EPSP) synthase (aroA) cloned from Campylobacter jejuni (Cj) strain 81116 was identified by complementation of an Escherichia coli (Ec) auxotrophic aroA mutant. The Cj aroA gene has been sequenced. It encodes an enzyme of 428 amino acids

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

    DEFF Research Database (Denmark)

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

    2000-01-01

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

  2. Predominant Campylobacter jejuni sequence types persist in Finnish chicken production.

    Directory of Open Access Journals (Sweden)

    Ann-Katrin Llarena

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

  3. Gene expression profiling of innate immune response to Campylobacter jejuni infection in the bursa of broilers

    Science.gov (United States)

    Campylobacter jejuni (C. jejuni) is a commensal microorganism in chickens, but caused significant health problems in humans. Reduction of C. jejuni colonization in the chicken gut will significantly decrease human campylobacteriosis. To study host response to C. jejuni infection in broilers, both ...

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

    NARCIS (Netherlands)

    Gaasbeek, E.J.; Wagenaar, J.A.; Guilhabert, M.R.; Wösten, M.M.S.M.; Putten, van J.P.M.; Graaf-van Bloois, van der L.; Parker, C.T.; Wal, van der F.J.

    2009-01-01

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

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

    DEFF Research Database (Denmark)

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

    Campylobacter jejuni is the primary food borne bacterial pathogen in the developed world. The most important reservoir for C. jejuni is the gut of chickens, which are colonized commensally and efficiently by this organism. Predominantly the mucus filled crypts of the lower gastrointestinal tract...... are found to be colonized by C. jejuni, and the bacteria are expected to be attracted to this particular environment by chemotaxis. In order to explore the role of chemotaxis in C. jejuni colonization we are construction deletion mutants in the putative chemoreceptors of the sequenced strain NCTC11168....... These mutants will be analyzed for their chemotatic capacity in order to investigate the chemoreceptor function and to identify matching chemoeffectors. Furthermore, selected mutants will be investigated for their ability to colonize chickens with focus on establishment, level, and persistence. Special emphasis...

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

    DEFF Research Database (Denmark)

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

    2009-01-01

    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......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......, 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...

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

    DEFF Research Database (Denmark)

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

    Campylobacter jejuni is the primary food borne bacterial pathogen in the developed world and the bacteria causes millions of gastroenteritis cases each year. The most important reservoir for C. jejuni is the gut of chickens, which are colonized commensally and efficiently by this organism....... Predominantly the mucus filled crypts of the lower gastrointestinal tract of chickens are found to be colonized by C. jejuni, and the bacteria are expected to be attracted to this particular environment by chemotaxis. From the full genome sequence of C. jejuni NCTC11168 several chemotactic proteins...... function and to identify matching chemoeffectors. Furthermore, selected mutants will be investigated for their ability to colonize chickens with focus on establishment, level, and persistence. Special emphasis will be held at characterizing the colonization of mucus layers....

  8. Genetic Characterization of Campylobacter Jejuni and C. coli Isolated From Broilers Using flaA PCR-Restriction Fragment Length Polymorphism Method in Shiraz, Southern Iran.

    Science.gov (United States)

    Khoshbakht, Rahem; Tabatabaei, Mohammad; Hosseinzadeh, Saeid; Shirzad Aski, Hesamaddin; Seifi, Saeed

    2015-05-01

    Thermophilic campylobacters, particularly Campylobacter jejuni and C. coli are the main agents of human campylobacteriosis. Campylobacter contaminated chicken products is the most important source of foodborne gastroenteritis. Evaluation of genetic diversity among Campylobacter population is critical for understanding the epidemiology of this bacterium and developing effective control strategies against Campylobacter infections and other related disorders. The aim of this study was to investigate the polymorphism of thermophilic Campylobacter isolated from broiler fecal samples in Shiraz, southern Iran. Ninety Campylobacter isolates were recovered from broiler feces using enrichment process followed by cultivation method. The isolates were species typing on the basis of polymerase chain reaction (PCR) detection of 16SrRNA and multiplex PCR for determining two thermophilic species. To evaluate strain diversity of thermophilic Campylobacter isolates, flaA PCR-Restriction Fragment Length Polymorphism (RFLP) was performed using DdeI restriction enzyme. All 90 Campylobacter isolates confirmed by m-PCR were successfully typed using flaA-PCR-RFLP. Eleven different types were defined according to flaA-typing method and the RFLP patterns were located at three separate clusters in RFLP image analysis dendrogram. Campylobacter jejuni isolates significantly showed more variety than C. coli isolates. A relatively low genetic diversity existed among C. jejuni and C. coli isolated from broilers in Shiraz, southern Iran. In our knowledge, this was the first report of genetic diversity among broiler originated human pathogen thermophilic campylobacters in Shiraz, southern Iran.

  9. Rapid Detection of Campylobacter jejuni by Polymerase Chain Reaction and Evaluation of its Sensitivity and Specificity

    OpenAIRE

    Razei A; Sorouri R; Aghamollaei H; Mousavi SL

    2017-01-01

    Introduction: Campylobacter jejuni is one of the most common causes of food poising in humans. Rapid and specific detection of these bacteria has an important role in diagnosis and treatment of infection. The aim of this study was to design a specific PCR for the detection of Campylobacter jejuni. Methods: In this experimental study, oxidoreductase gene from the Campylobacter jejuni was selected for rapid and specific detection. For this purpose, specific primers were designed and charecte...

  10. Tetracycline Resistance Genes in Campylobacter jejuni and C. coli Isolated From Poultry Carcasses.

    Science.gov (United States)

    Abdi-Hachesoo, Bahman; Khoshbakht, Rahem; Sharifiyazdi, Hassan; Tabatabaei, Mohammad; Hosseinzadeh, Saeid; Asasi, Keramat

    2014-09-01

    Campylobacter is one of the leading bacterial species causing foodborne illnesses in humans. Antimicrobial agents have been extensively used for treatment of Campylobacter infections; but in the recent years, both animal and human isolates of this bacterium have shown resistance to several antibiotics such as tetracycline. The aim of this study was to investigate the presence of genetic determinants of tetracycline resistance in Campylobacter spp. recovered from poultry carcasses in Shiraz, Iran. Eighty-three thermophilic Campylobacter spp. Isolates were first identified based on multiplex polymerase chain reaction (PCR) and then screened for presence of tetracycline resistance genes (tet (A), tet (B), tet (O) and te (S)) by PCR. The overall prevalence of Campylobacter jejuni and C. coli among the examined isolates was 51.8% and 48.2%, respectively. Tetracycline resistance genes of tet (B) and tet (S) were not seen among these Campylobacter spp. Isolates, whereas the most common tet gene identified was tet (O), found in 83.1% (69/83) of all the isolates. The tet (O) gene sequence comparison between C. jejuni and C. coli showed 100% similarity and these sequences (JX853721and JX853722) were also identical to the homologous sequences of other strains of Campylobacter spp. existing in the GenBank databases. In addition, tet (A) was found in 18% (15/83) of Campylobacter spp. isolates. To our knowledge, this represents the first report of tet (A) in Campylobacter spp. There was 100% homology between the sequences of tet (A) from this study (JX891463 and JX891464) and the tet (A) sequences mentioned for other bacteria in the GenBank databases. The high prevalence of tet (O) resistance gene along with new detection of tet (A) resistance gene in Campylobacter spp. isolated from poultry carcasses revealed an extensive tetracycline resistance among Campylobacter isolates from poultry in Iran. It emphasized the need for cautious use of tetracycline in poultry production to

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

    Science.gov (United States)

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

    2015-01-01

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

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

    Directory of Open Access Journals (Sweden)

    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.

  13. Antimicrobial activities of isothiocyanates against Campylobacter jejuni isolates

    Directory of Open Access Journals (Sweden)

    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.

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

    NARCIS (Netherlands)

    Doorduyn, Y.; Brandhof, van den W.E.; Duynhoven, van Y.T.H.P.; Breukink, B.J.; Wagenaar, J.A.; Pelt, van W.

    2010-01-01

    A case-control study comprising 1315 Campylobacter jejuni cases, 121 Campylobacter coli cases and 3409 frequency-matched controls was conducted in The Netherlands in 2002-2003. Risk factors for both C. jejuni and C. coli enteritis were consumption of undercooked meat and barbecued meat, ownership of

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

    Science.gov (United States)

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

    2016-04-15

    A quartz crystal microbalance (QCM) sensor platform was used to develop an immunosensor for the detection of food pathogen Campylobacter jejuni. Rabbit polyclonal antibodies and commercially available mouse monoclonal antibodies against C. jejuni were investigated to construct direct, sandwich and gold-nanoparticles (AuNPs) amplified sandwich assays. The performance of the QCM immunosensor developed using sandwich assay by utilising the rabbit polyclonal antibody as the capture antibody and conjugated to AuNPs as the detection antibody gave the highest sensitivity. This sensor achieved a limit of detection (LOD) of 150 colony forming unit (CFU)mL(-1) of C. jejuni in solution. The QCM sensor showed excellent sensitivity and specificity for Campylobacter detection with low cross reactivity for other foodborne pathogens such as Salmonella Typhimurium, (7%) Listeria monocytogenes (3%) and Escherichia coli (0%). The development of this biosensor would help in the sensitive detection of Campylobacter which can result in reducing pre-enrichment steps; hence, reducing assay time. This work demonstrates the potential of this technology for the development of a rapid and sensitive detection method for C. jejuni. Copyright © 2015 Elsevier B.V. All rights reserved.

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

    Science.gov (United States)

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

    2015-04-01

    In order to compare the prevalence of Campylobacter coli and Campylobacter jejuni during the processing of broilers at slaughterhouse a total of 848 samples were analyzed during 2012 in southern Spain. Four hundred and seventy six samples were collected from cloaca, carcass surfaces and quartered carcasses. Moreover, 372 environmental swabs from equipment and scalding water were collected. Minimum inhibitory concentration (MIC) to ciprofloxacin, erythromycin, streptomycin, tetracycline and gentamicin was determined for isolates from chicken meat. The general prevalence of Campylobacter was 68.8% (40.2% of C. coli and 28.5% of C. jejuni). The relative prevalence of C. coli increased from loading dock area (41.5%) to packing area (64.6%). In contrast, the relative prevalence of C. jejuni decreased from 58.5% to 35.4%. These differences between species from initial to final area were significant (p=0.02). The highest antimicrobial resistance for C. jejuni and C. coli was detected to tetracycline (100%) and ciprofloxacin (100%), respectively. Campylobacter coli showed an antimicrobial resistance significantly higher than C. jejuni to streptomycin (p=0.002) and erythromycin (p<0.0001). Copyright © 2015 Elsevier Ltd. All rights reserved.

  17. Detection and quantification of Campylobacter jejuni and Campylobacter coli using real-time multiplex PCR.

    Science.gov (United States)

    Toplak, N; Kovač, M; Piskernik, S; Možina, S Smole; Jeršek, B

    2012-04-01

    We describe a real-time quantitative multiplex polymerase chain reaction (qmPCR) assay to identify and discriminate between isolates of Campylobacter jejuni and Campylobacter coli. Two novel sets of primers and hydrolysis probes were designed to amplify the unique DNA sequences within the hipO, ccoN and cadF genes that are specific to Camp. jejuni and Camp. coli. Using the designed optimized qmPCR assay conditions, the amplification efficiency is in range from 108 to 116%. These qmPCR assays are highly specific for Camp. jejuni and Camp. coli, as seen through testing of 40 Campylobacter strains and 17 non-Campylobacter strains. In chicken juice and tap water models spiked with known quantities of Camp. jejuni, qmPCR detected 10(2) -10(3) CFU ml(-1) within 4 h. The qmPCR assays developed in this study provide reliable and simultaneous detection and quantification of Camp. jejuni and Camp. coli, with good amplification reaction parameters. Following further validation, the qmPCR assay reported here has the potential to be applied to various sample types as an alternative and rapid methodology. © 2012 The Authors. Journal of Applied Microbiology © 2012 The Society for Applied Microbiology.

  18. Comparative genomic fingerprinting for the subtyping of Campylobacter jejuni and Campylobacter coli biotypes

    Directory of Open Access Journals (Sweden)

    Miljković-Selimović Biljana

    2017-01-01

    Full Text Available Introduction/Objective. Thermophilic campylobacters, especially Campylobacter jejuni (C. jejuni and Campylobacter coli (C. coli, are the most important causes of bacterial diarrhea in developed and developing countries. The disease can occur as a sporadic infection or as large and small outbreaks. Phenotyping and genotyping methods are in use to determine similarities between strains as well their possible common origin. The goal of the study was to compare discriminatory power of biotyping tests and comparative genomic fingerprinting (CGF 40 (100%, as well as a combination of the two tests in detection of clonality or epidemiological relatedness between the studied strains. Methods. We investigated 23 Campylobacter strains using biotyping and CGF typing. Results. We found that biotyping was a more discriminatory method for C. coli, and CGF for C. jejuni strains. In the discrimination of C. jejuni strains, CGF had better discriminatory power [Simpson’s index of diversity (ID was 0.879] over the discrimination of C. coli strains (Simpson’s ID was 0.389. Conclusion. Biotyping and CGF can be complementary methods in detection of similarity, relatedness and possible common origin between strains since the combination of biotyping and CGF methods gives more precise data about diversity within C. coli and C. jejuni strains. [Project of the Serbian Ministry of Education, Science and Technological Development, Grant no. TR34008

  19. Campylobacter jejuni Actively Invades the Amoeba Acanthamoeba polyphaga and Survives within Non Digestive Vacuoles

    Science.gov (United States)

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

    2013-01-01

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

  20. Characterization of the Campylobacter jejuni population in the barnacle geese reservoir.

    Science.gov (United States)

    Llarena, A-K; Skarp-de Haan, C P A; Rossi, M; Hänninen, M-L

    2015-05-01

    Campylobacter spp. are the most common cause of bacterial gastroenteritis worldwide and have been isolated from a wide number of different hosts and environmental sources. Waterfowl is considered a natural reservoir for this zoonotic bacterium and may act as a potential infection source for human campylobacteriosis. In this study, faecal samples from 924 barnacle geese were tested for the presence of C. jejuni and C. coli. The resulting C. jejuni and C. coli populations were characterized by multilocus sequence typing (MLST), structure analysis by BAPS and phylogenetic analysis based on full genome sequences. The prevalences of C. jejuni in barnacle geese faeces were 11.5% and 23.1% in 2011 and 2012, respectively, and only 0.2% of the samples were positive for C. coli in both years. Furthermore, a possible adaption of the clonal complexes (CCs) ST-702 and ST-1034 to the barnacle geese reservoir was found, as these two CCs represented the majority of the typed isolates and were repeatedly isolated from different flocks at several time-points. Further core genome phylogenetic analysis using ClonalFrame revealed a formation of a distinct monophyletic lineage by these two CCs, suggesting a certain degree of clonality of the C. jejuni population adapted to barnacle geese. Therefore, although STs also commonly found in humans patients (e.g. ST-45) were among the barnacle geese C. jejuni isolates, this reservoir is probably an infrequent source for human campylobacteriosis. © 2014 Blackwell Verlag GmbH.

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

    Directory of Open Access Journals (Sweden)

    Jenny Olofsson

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

  2. Whole-Genome Sequencing in Epidemiology of Campylobacter jejuni Infections.

    Science.gov (United States)

    Llarena, Ann-Katrin; Taboada, Eduardo; Rossi, Mirko

    2017-05-01

    This review describes the current state of knowledge regarding the application of whole-genome sequencing (WGS) in the epidemiology of Campylobacter jejuni , the leading cause of bacterial gastroenteritis worldwide. We describe how WGS has increased our understanding of the evolutionary and epidemiological dynamics of this pathogen and how WGS has the potential to improve surveillance and outbreak detection. We have identified hurdles to the full implementation of WGS in public health settings. Despite these challenges, we think that ample evidence is available to support the benefits of integrating WGS into the routine monitoring of C. jejuni infections and outbreak investigations. Copyright © 2017 American Society for Microbiology.

  3. Prevalence of Campylobacter jejuni and Campylobacter coli among broilers in Bareilly region

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    Hina Malik

    2014-10-01

    Full Text Available Aim: To determine the prevalence of Campylobacter jejuni and Campylobacter coli among broilers at the time of slaughter in and around Bareilly, India. Materials and Methods: A total of 100 chicken caecal samples were screened by conventional plating in modified charcoal cefoperazone deoxycholate agar with incubation at 42°C for 48 h under microaerophilic conditions. The characteristic colonies were confirmed by morphological and biochemical characteristics and multiplex polymerase chain reaction (mPCR assay targeting lpxA gene. Results: Out of 100 chicken caecal samples, 32 yielded isolates with typical phenotypic of Campylobacter species. The hippurate hydrolysis test found to be positive for 2 isolates, categorized as C. jejuni and negative for 30 isolates. The mPCR assay targeting lpxA gene also confirmed 2 (6.25% isolates as C. jejuni, and 30 (93.75% isolates as C. coli. Conclusion: The present study showed broilers to an important source of Campylobacter in the region with predominance of C. coli than C. jejuni indicating a shift in the prevalence of important species of Campylobacter. To understand the variation in pattern of occurrence of species with high prevalence of organisms, detail studies on the ecology of campylobacteriosis are suggested.

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

    DEFF Research Database (Denmark)

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

    2016-01-01

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

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

    DEFF Research Database (Denmark)

    Pedersen, Karl; Wedderkopp, A.

    2003-01-01

    Aims : To investigate the prevalence of quinolone resistance among Campylobacter jejuni and Camp. coli isolates from Danish poultry at the farm level, as well as for the whole country. Methods and Results : Data and isolates were collected from a national surveillance of Campylobacter in poultry......-resistant variant. Conclusions : Overall, quinolone resistance among Campylobacter isolates from Danish broilers was 7.5% in 1998 and 1999; it was higher among Camp. coli than Camp. jejuni . Genetic diversity among resistant isolates was lower than among susceptible isolates, and certain clones existed in both...... a resistant and a susceptible variant. Some resistant clones appeared to persist on the farms and were repeatedly isolated from poultry flocks. Significance and Impact of the Study : The study is important for the understanding of persistence and dynamics of Campylobacter in broiler houses. It also highlights...

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

    Directory of Open Access Journals (Sweden)

    Tambur Zoran Ž.

    2010-01-01

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

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

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    Hartley Lauren E

    2009-06-01

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

  8. Characterization of Campylobacter jejuni and Campylobacter coli Broiler Isolates by Whole-Genome Sequencing

    DEFF Research Database (Denmark)

    Cantero, Guillermo; Correa-Fiz, Florencia; Ronco, Troels

    2017-01-01

    Campylobacter has been the most commonly reported cause of bacterial diarrheal disease in humans in the European Union since 2005. Most broiler batches at slaughter are colonized with Campylobacter, and the major source of infection is contaminated poultry meat. The aim of this study was to chara......Campylobacter has been the most commonly reported cause of bacterial diarrheal disease in humans in the European Union since 2005. Most broiler batches at slaughter are colonized with Campylobacter, and the major source of infection is contaminated poultry meat. The aim of this study...... was to characterize a selection of Campylobacter jejuni and Campylobacter coli isolates from broilers through whole-genome sequencing (WGS). A total of 16 isolates (C. jejuni = 12 and C. coli = 4) from five broiler farms from Catalonia (northeastern Spain) were analyzed. A phylogenetic analysis based on 8420 single...

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

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    Christina S Vegge

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

  10. Antimicrobial resistance in Campylobacter coli and Campylobacter jejuni in cynomolgus monkeys (Macaca fascicularis) and eradication regimens.

    Science.gov (United States)

    Koga, Tetsufumi; Aoki, Wataru; Mizuno, Takashi; Wakazono, Kuniko; Ohno, Junki; Nakai, Tsunehiro; Nomiya, Takao; Fujii, Miki; Fusegawa, Keiichi; Kinoshita, Kazuya; Hamada, Takakazu; Ikeda, Yoshinori

    2017-02-01

    Campylobacter spp. are zoonotic pathogens, however, knowledge about their presence and antimicrobial resistance in nonhuman primates is limited. Our animal facility purchased cynomolgus monkeys (Macaca fascicularis) from various Asian countries: China, Cambodia, Indonesia, the Philippines, and Vietnam. Colonization by Campylobacter spp. was investigated in 238 of the monkeys from 2009 to 2012 and antimicrobial susceptibility testing was carried out for these isolates. Furthermore, we eradicated these pathogens from these monkeys. Campylobacter spp. were isolated from 47 monkeys from three specific countries: China, Cambodia, and Indonesia, with respective isolation rates of 15%, 36%, and 67%. Two monkeys, which were each infected with Campylobacter jejuni and Campylobacter coli, showed clinical symptoms of diarrhea and bloody feces. In total, 41 isolates of C. coli and 17 isolates of C. jejuni were detected. Antimicrobial susceptibility varied: in the monkeys from China, erythromycin (ERY)-, tetracycline (TET)-, and ciprofloxacin-resistant C. coli, in the monkeys from Cambodia, amoxicillin-intermediate, TET- and ciprofloxacin-resistant C. coli and amoxicillin- and ciprofloxacin-resistant C. jejuni, and in the monkeys from Indonesia, ciprofloxacin-resistant C. coli and TET- and ciprofloxacin-resistant C. jejuni were common (>75%). Multiresistant isolates of C. coli were found in monkeys from all countries and multiresistant isolates of C. jejuni were found in monkeys from Indonesia. The eradication rate with azithromycin was comparable to that with gentamicin (GEN) by oral administration, and was higher than those with amoxicillin-clavulanic acid (AMC) and chloramphenicol (CHL). From the perspective of zoonosis, we should acknowledge multiresistant Campylobacter spp. isolated from the monkeys as a serious warning. Copyright © 2015. Published by Elsevier B.V.

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

    Energy Technology Data Exchange (ETDEWEB)

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

    1987-02-01

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

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

    International Nuclear Information System (INIS)

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

    1987-01-01

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

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

    NARCIS (Netherlands)

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

    2000-01-01

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

  14. Investigation of Campylobacter jejuni and Campylobacter coli isolated from broilers susceptibility to antibiotics

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    Tambur Zoran

    2015-01-01

    Full Text Available In this work sensitivity to eritromycine, tetracycline and ciprofloxacin was investigated on 16 (sixteen strains Campylobacter jejuni and Campylobacter coli originated from broilers’ coecum by use of dics-diffussion method and E-test. Out of 10 (ten examined strains of C. jejuni, 1 strain (10% was resistant to erithromycin, 5 (five examined strains (50% to tetracycline and 4 (four examined strains (40% to ciprofloxacine. Out of 6 investigated strains of C. coli, 1 strain was resistant to erithromycin (16,7%, 5 of investigated strains to tetracycline and ciprofloxacine (83,3% (table 1 and 2, graph 3. Out of 16 (sixteen investigated strains of C. jejuni and C. coli, 9 (nine strains were resistant to ciprofloxacine (56,2%, 2 (two strains to erythromycine (12,5% and 10 of investigated strains to tetracycline (62,5%.

  15. The Prevalence of Antibiotic and Biocide Resistance Among Campylobacter coli and Campylobacter jejuni from Different Sources

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    Ana Mavri

    2012-01-01

    Full Text Available The increasing levels of antimicrobial resistance among foodborne bacteria are recognised as an important emerging public health problem. Reduced susceptibility to biocides also appears to be increasing. A potential concern is the possibility that the widespread use of biocides is responsible for the selection and maintenance of antibiotic-resistant bacteria. Here, we examine the prevalence of erythromycin, ciprofloxacin, triclosan, benzalkonium chloride, chlorhexidine diacetate, cetylpyridinium chloride, trisodium phosphate and sodium dodecyl sulphate resistance among 27 isolates of Campylobacter coli and 15 isolates of Campylobacter jejuni from food, animal, human and environmental water sources. These antimicrobial susceptibilities were determined by the broth microdilution method. In the 42 Campylobacter strains studied, different antibiotic resistance levels were seen. The resistance to erythromycin and ciprofloxacin was observed in 14.3 % of Campylobacter strains. A higher rate of erythromycin resistance and multi-resistance was observed among isolated C. coli than among C. jejuni strains. Similar situations were seen for triclosan. Conversely, the level of benzalkonium chloride resistance was higher in C. jejuni than in C. coli. No correlation between biocide and antibiotic resistance was observed. This study does not provide evidence to confirm that tolerance to biocides is connected to antibiotic resistance in Campylobacter.

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

    Science.gov (United States)

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

    2014-04-01

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

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

    Science.gov (United States)

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

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

    Science.gov (United States)

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

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

    DEFF Research Database (Denmark)

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

    2014-01-01

    Previous studies have identified a specific modification of the capsular polysaccharide as receptor for phages that infect Campylobacter jejuni. Using acapsular kpsM mutants of C. jejuni strains NCTC11168 and NCTC12658, we found that bacteriophage F341 infects C. jejuni independently of the capsule...

  20. Microbiota-Derived Short-Chain Fatty Acids Modulate Expression of Campylobacter jejuni Determinants Required for Commensalism and Virulence.

    Science.gov (United States)

    Luethy, Paul M; Huynh, Steven; Ribardo, Deborah A; Winter, Sebastian E; Parker, Craig T; Hendrixson, David R

    2017-05-09

    Campylobacter jejuni promotes commensalism in the intestinal tracts of avian hosts and diarrheal disease in humans, yet components of intestinal environments recognized as spatial cues specific for different intestinal regions by the bacterium to initiate interactions in either host are mostly unknown. By analyzing a C. jejuni acetogenesis mutant defective in converting acetyl coenzyme A (Ac-CoA) to acetate and commensal colonization of young chicks, we discovered evidence for in vivo microbiota-derived short-chain fatty acids (SCFAs) and organic acids as cues recognized by C. jejuni that modulate expression of determinants required for commensalism. We identified a set of C. jejuni genes encoding catabolic enzymes and transport systems for amino acids required for in vivo growth whose expression was modulated by SCFAs. Transcription of these genes was reduced in the acetogenesis mutant but was restored upon supplementation with physiological concentrations of the SCFAs acetate and butyrate present in the lower intestinal tracts of avian and human hosts. Conversely, the organic acid lactate, which is abundant in the upper intestinal tract where C. jejuni colonizes less efficiently, reduced expression of these genes. We propose that microbiota-generated SCFAs and lactate are cues for C. jejuni to discriminate between different intestinal regions. Spatial gradients of these metabolites likely allow C. jejuni to locate preferred niches in the lower intestinal tract and induce expression of factors required for intestinal growth and commensal colonization. Our findings provide insights into the types of cues C. jejuni monitors in the avian host for commensalism and likely in humans to promote diarrheal disease. IMPORTANCE Campylobacter jejuni is a commensal of the intestinal tracts of avian species and other animals and a leading cause of diarrheal disease in humans. The types of cues sensed by C. jejuni to influence responses to promote commensalism or

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

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    Metris Aline

    2011-11-01

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

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

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    Banya Banowary

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

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

    Science.gov (United States)

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

    2015-01-01

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

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

    Science.gov (United States)

    2011-05-01

    Society for Microbiology. All Rights Reserved. Discrimination of Major Capsular Types of Campylobacter jejuni by Multiplex PCR’Vt Frederic Poly...two PCRs with sensitivities and specificities ranging from 90 tn 100% using 244 strains of knnwn Penner type. Campylobacter jejwzi is one of the...2. REPORT TYPE 3. DATES COVERED 00-00-2011 to 00-00-2011 4. TITLE AND SUBTITLE Discrimination of Major Capsular Types of Campylobacter jejuni

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

    Directory of Open Access Journals (Sweden)

    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.

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

    Directory of Open Access Journals (Sweden)

    Muhammad A Javed

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

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

    Science.gov (United States)

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

    2013-11-25

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

  8. Role of environmental survival in transmission of Campylobacter jejuni.

    Science.gov (United States)

    Bronowski, Christina; James, Chloe E; Winstanley, Craig

    2014-07-01

    Campylobacter species are the most common cause of bacterial gastroenteritis, with C. jejuni responsible for the majority of these cases. Although it is clear that livestock, and particularly poultry, are the most common source, it is likely that the natural environment (soil and water) plays a key role in transmission, either directly to humans or indirectly via farm animals. It has been shown using multilocus sequence typing that some clonal complexes (such as ST-45) are more frequently isolated from environmental sources such as water, suggesting that strains vary in their ability to survive in the environment. Although C. jejuni are fastidious microaerophiles generally unable to grow in atmospheric levels of oxygen, C. jejuni can adapt to survival in the environment, exhibiting aerotolerance and starvation survival. Biofilm formation, the viable but nonculturable state, and interactions with other microorganisms can all contribute to survival outside the host. By exploiting high-throughput technologies such as genome sequencing and RNA Seq, we are well placed to decipher the mechanisms underlying the variations in survival between strains in environments such as soil and water and to better understand the role of environmental persistence in the transmission of C. jejuni directly or indirectly to humans. © 2014 The Authors. FEMS Microbiology Letters published by John Wiley & Sons Ltd on behalf of Federation of European Microbiological Societies.

  9. Bacteriophage F336 recognizes the capsular phosphoramidate modification of Campylobacter jejuni NCTC11168.

    Science.gov (United States)

    Sørensen, Martine C Holst; van Alphen, Lieke B; Harboe, Anne; Li, Jianjun; Christensen, Bjarke Bak; Szymanski, Christine M; Brøndsted, Lone

    2011-12-01

    Bacteriophages infecting the food-borne human pathogen Campylobacter jejuni could potentially be exploited to reduce bacterial counts in poultry prior to slaughter. This bacterium colonizes the intestinal tract of poultry in high numbers, and contaminated poultry meat is regarded as the major source of human campylobacteriosis. In this study, we used phage F336 belonging to the Myoviridae family to select a C. jejuni NCTC11168 phage-resistant strain, called 11168R, with the aim of investigating the mechanisms of phage resistance. We found that phage F336 has reduced adsorption to 11168R, thus 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 MeOPN transferase, contains a tract of 10 Gs, resulting in a nonfunctional gene product. However, when 11168R reverted back to phage sensitive, cj1421 contained 9 Gs, and the GalfNAc MeOPN was regained in this strain. In summary, we have identified the phase-variable MeOPN moiety, a common component of the diverse capsular polysaccharides of C. jejuni, as a novel receptor of phages infecting this bacterium.

  10. Prevalence of Thermotolerant Campylobacter spp. in Chicken Meat in Croatia and Multilocus Sequence Typing of a Small Subset of Campylobacter jejuni and Campylobacter coli Isolates

    Directory of Open Access Journals (Sweden)

    Andrea Humski

    2016-01-01

    Full Text Available In order to detect thermotolerant Campylobacter spp., 241 samples of fresh chicken meat, at retail in Croatia, were analysed according to a standard method, followed by biochemical test and molecular polymerase chain reaction/restriction enzyme analysis for exact species determination. Campylobacter spp. prevalence was 73.86 %. Campylobacter jejuni and Campylobacter coli were isolated from 53.53 and 15.35 % of the samples, respectively. In 4.98 % of isolates thermotolerant Campylobacter spp. were not determined. The multi locus sequence typing method was used to evaluate genetic diversity of eight Campylobacter jejuni and four Campylobacter coli isolates. To our knowledge, these results of genotyping provided the first data on the presence of sequence types (STs and clonal complexes (CCs of Campylobacter jejuni and C. coli isolates in Croatia. By applying the multilocus sequence typing, a new allele of tkt gene locus was discovered and marked tkt508. The C. jejuni ST 6182 and C. coli ST 6183 genotypes were described for the fi rst time, and all other identified genotypes were clustered in the previously described sequence types and clonal complexes. These findings provide useful information on the prevalence and epidemiology of Campylobacter jejuni and C. coli in Croatia.

  11. [Microbiological diagnosis of infections caused by Campylobacter jejuni and Campylobacter coli in humans].

    Science.gov (United States)

    Rokosz, Natalia; Rastawicki, Waldemar; Wołkowicz, Tomasz

    2014-01-22

    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.

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

    Directory of Open Access Journals (Sweden)

    Natalia Rokosz

    2014-01-01

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

  13. Effect of chicken meat environment on gene expression of Campylobacter jejuni and its relevance to survival in food.

    Science.gov (United States)

    Ligowska, Małgorzata; Cohn, Marianne Thorup; Stabler, Richard A; Wren, Brendan W; Brøndsted, Lone

    2011-03-01

    Poultry meat is the major food source responsible for gastrointestinal infections caused by the human pathogen Campylobacter jejuni. Even though C. jejuni does not grow below 30 °C, the bacterium survives on raw meat surfaces at refrigerated temperatures and thus poses a risk to the consumer. Previously, we have shown that chicken meat juice prolongs survival of C. jejuni at 5 °C compared to laboratory medium, suggesting that compounds present in meat juice influence adaptation to low temperatures. In the present study we have used chicken meat juice to identify C. jejuni genes that are differentially expressed in a typical chicken meat environment encountered by consumers. The analysis showed that chicken meat juice increased expression of luxS involved in quorum sensing, as well as a gene involved in O-linked flagellin glycosylation in C. jejuni, while expression of haemin uptake and the peroxide stress response genes were reduced. Furthermore, we propose that LuxS may play a key role in adaptation to the chicken meat juice environment, as lack of the luxS gene reduces the ability of C. jejuni to survive in chicken meat juice at low temperature. Finally, our data suggest that part of an ABC transport system is induced and we speculate that uptake of cryoprotectants may be important for C. jejuni to adapt to low temperature. In summary, we found that C. jejuni has a specific but limited transcriptional response to chicken meat juice and that luxS has an impact on the prolonged survival of C. jejuni in this important environment in the food chain. Copyright © 2010 Elsevier B.V. All rights reserved.

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

    Directory of Open Access Journals (Sweden)

    Jonas Waldenström

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

  15. Rapid Detection of Campylobacter jejuni by Polymerase Chain Reaction and Evaluation of its Sensitivity and Specificity

    Directory of Open Access Journals (Sweden)

    Razei A

    2017-06-01

    Full Text Available Introduction: Campylobacter jejuni is one of the most common causes of food poising in humans. Rapid and specific detection of these bacteria has an important role in diagnosis and treatment of infection. The aim of this study was to design a specific PCR for the detection of Campylobacter jejuni. Methods: In this experimental study, oxidoreductase gene from the Campylobacter jejuni was selected for rapid and specific detection. For this purpose, specific primers were designed and charecterized by bioinformatics software. Bacterial genome was extracted by phenol-chloroform method and PCR was optimized to obtain a specific product. Specificity of the designed reaction was investigated using six bacterial species. The sensitivity of the PCR reaction was calculated by the serial dilutions method. Results: The designed primer was specific to oxidoreductase gene of Campylobacter jejuni and after optimization, a unique 167-bp band was amplified. This primer was specific to Campylobacter jejuni and did not show any cross reaction with other bacterial genomes. The detection limit of this reaction was 5 pg of genomic DNA. Conclusions: The optimized PCR used in this study was a rapid, sensitive, and specific method for detection of Campylobacter jejuni. For confirmation of this method, detection of C. jejuni from food samples is proposed.

  16. High-resolution genomic fingerprinting of Campylobacter jejuni and Campylobacter coli by analysis of amplified fragment length polymorphisms

    DEFF Research Database (Denmark)

    Kokotovic, Branko; On, Stephen L.W.

    1999-01-01

    A method for high-resolution genomic fingerprinting of the enteric pathogens Campylobacter jejuni and Campylobacter coli, based on the determination of amplified fragment length polymorphism, is described. The potential of this method for molecular epidemiological studies of these species...... to available epidemiological data. We conclude that this amplified fragment length polymorphism fingerprinting method may be a highly effective tool for molecular epidemiological studies of Campylobacter spp....

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

    Science.gov (United States)

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

    2016-09-15

    Campylobacter is the most common causative agent of human bacterial gastroenteritis and is frequently found in surface water, where it indicates recent contamination with animal faeces, sewage effluent, and agricultural run-off. The contribution of different animal reservoirs to surface water contamination with Campylobacter is largely unknown. In the Netherlands, the massive poultry culling to control the 2003 avian influenza epidemic coincided with a 44-50% reduction in human campylobacteriosis cases in the culling areas, suggesting substantial environment-mediated spread of poultry-borne Campylobacter. We inferred the origin of surface water Campylobacter jejuni and Campylobacter coli strains in Luxembourg and the Netherlands, as defined by multilocus sequence typing, by comparison to strains from poultry, pigs, ruminants, and wild birds, using the asymmetric island model for source attribution. Most Luxembourgish water strains were attributed to wild birds (61.0%), followed by poultry (18.8%), ruminants (15.9%), and pigs (4.3%); whereas the Dutch water strains were mainly attributed to poultry (51.7%), wild birds (37.3%), ruminants (9.8%), and pigs (1.2%). Attributions varied over seasons and surface water types, and geographical variation in the relative contribution of poultry correlated with the magnitude of poultry production at either the national or provincial level, suggesting that environmental dissemination of Campylobacter from poultry farms and slaughterhouses can be substantial in poultry-rich regions. Copyright © 2016 Elsevier Ltd. All rights reserved.

  18. Rapid detection and differentiation of Campylobacter jejuni, Campylobacter coli, and Campylobacter lari in food, using multiplex real-time PCR.

    Science.gov (United States)

    Mayr, A M; Lick, S; Bauer, J; Thärigen, D; Busch, U; Huber, I

    2010-02-01

    A multiplex real-time PCR assay based on four differently labeled TaqMan probes for detection and differentiation of the thermophilic Campylobacter species C. jejuni, C. coli, and C. lari was established and validated in food products. This assay combines two previously published PCR assays for C. jejuni and C. coli with a newly developed detection assay for C. lari and an internal amplification control system. The selectivity of the method was determined by analyzing 70 Campylobacter strains and 43 strains of other bacteria. The sensitivity was 50 fg of C. jejuni and C. lari DNA and 500 fg of C. coli DNA per PCR. It was possible to detect 1 to 10 CFU/25 g of food before preenrichment of all three species. More than 400 samples of various foods (poultry, seafood, and meat) were analyzed after 48 h of preenrichment parallel to the conventional diagnostic method of culture and biochemical identification. Using the established real-time PCR assay, 55.4% of the samples were recognized as positive for thermophilic Campylobacter species, whereas with the conventional method only 40.3% of the samples were positive. The real-time PCR assay also detected contaminations with two different Campylobacter species in 32.6% of the analyzed poultry samples, a finding of epidemiological interest. Compared with the original PCR method, which was established for the differentiation of bacterial isolates of C. jejuni and C. coli, this new method also detects and distinguishes C. lari, was validated as an analytical tool for food analysis, and provides reliable and extensive results within 2 days.

  19. Mosaic Structure of a Multiple-Drug-Resistant, Conjugative Plasmid from Campylobacter jejuni

    National Research Council Canada - National Science Library

    Nirdnoy, Warawadee; Mason, Carl J; Guerry, Patricia

    2005-01-01

    Partial sequence analysis of a tet(O) plasmid from a multiple-drug-resistant clinical isolate of Campylobacter jejuni revealed 10 genes or pseudogenes encoding different aminoglycoside inactivating enzymes, transposaselike genes...

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

    Science.gov (United States)

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

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

    Science.gov (United States)

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

    2013-05-01

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

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

    Science.gov (United States)

    Flint, Annika; Sun, Yi-Qian

    2012-01-01

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

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

    DEFF Research Database (Denmark)

    Birk, Tina; Rosenquist, Hanne; Knøchel, Susanne

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

  4. Intrinsic and Extrinsic Aspects on Campylobacter jejuni Biofilms

    Directory of Open Access Journals (Sweden)

    Roberta T. Melo

    2017-07-01

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

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

    Science.gov (United States)

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

    2014-03-26

    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.

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

    DEFF Research Database (Denmark)

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

    Campylobacter jejuni is the primary food borne bacterial pathogen in the developed world. The most important reservoir for C. jejuni is the gut of chickens, which are colonized commensally and efficiently by this organism. Predominantly the mucus filled crypts of the lower gastrointestinal tract...... will be held at characterizing the colonization of mucus layers....

  7. Characterization of the specific interaction between sialoadhesin and sialylated Campylobacter jejuni lipooligosaccharides.

    NARCIS (Netherlands)

    Heikema, A.P.; Bergman, M.P.; Richards, H.; Crocker, P.R.; Gilbert, M.; Samsom, J.N.; Wamel, W.J.; Endtz, H.Ph.; van Belkum, A.

    2010-01-01

    In Campylobacter jejuni-induced Guillain-Barré syndrome (GBS), molecular mimicry between C. jejuni lipooligosaccharide (LOS) and host gangliosides leads to the production of cross-reactive antibodies directed against the peripheral nerves of the host. Currently, the presence of surface exposed

  8. Campylobacter jejuni translocation across intestinal epithelial cells is facilitated by ganglioside-like lipooligosaccharide structures

    NARCIS (Netherlands)

    R.P.L. Louwen (Rogier); E.E.S. Nieuwenhuis (Edward); L. van Marrewijk (Leonie); D. Horst-Kreft (Deborah); L.F. de Ruiter (Lilian); A.P. Heikema (Astrid); W.J.B. van Wamel (Willem); J.A. Wagenaar (Jaap); H.P. Endtz (Hubert); J.N. Samsom (Janneke); P. van Baarlen (Peter); A.S. Akhmanova (Anna); A.F. van Belkum (Alex)

    2012-01-01

    textabstractTranslocation across intestinal epithelial cells is an established pathogenic feature of the zoonotic bacterial species Campylobacter jejuni. The number of C. jejuni virulence factors known to be involved in translocation is limited. In the present study, we investigated whether

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

    Science.gov (United States)

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

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

    Science.gov (United States)

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

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

    NARCIS (Netherlands)

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

    2010-01-01

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

  12. Whole-genome sequence of the bacteriophage-sensitive strain Campylobacter jejuni NCTC12662

    DEFF Research Database (Denmark)

    Gencay, Yilmaz Emre; Sørensen, Martine C.H.; Brøndsted, Lone

    2017-01-01

    Campylobacter jejuni NCTC12662 has been the choice bacteriophage isolation strain due to its susceptibility to C. jejuni bacteriophages. This trait makes it a good candidate for studying bacteriophage-host interactions. We report here the whole-genome sequence of NCTC12662, allowing future...

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

    DEFF Research Database (Denmark)

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

    2011-01-01

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

  14. Phenotypic and Genotypic Detection of Campylobacter jejuni at Local Chicken and Chicken Meat

    Directory of Open Access Journals (Sweden)

    A Rosyidi

    2010-05-01

    Full Text Available The Objective of this study was to identify the existence of Campylobacter jejuni based on phenotypic and genotypic characteristic in local chicken and chicken meats. Samples of local chicken intestine and meat were tested for the bacterial existence. Phenotypic examination was carried out by means of cultivation followed by gram staining and biochemical tests. Genotypic examination was conducted by polymerase chain reaction (PCR using genus specific16S rRNA gene at 816 bp and membrane-associated protein A (mapA gene at 589 bp as Campylobacter jejuni species-specific gene. The result of phenotypic detection revealed the existence of Campylobacter spp as gram negative, curved rod shape, oxidase positive, urease negative and motile. Genotypic examination also indicated the existence of bacteria using both primers. However, no Campylobacter jejuni detected from meat of the chickens. The results suggest that the method of PCR using a primer detecting species-specific gene of Campylobacter jejuni gives a rapid and accurate detection of the bacteria as compared to that using phenotypic and biochemical test. Identification of Campylobacter spp from chicken meats should be improved with enrichment method and sample collection. (Animal Production 12(2: 128-134 (2010Key Words: Campylobacter jejuni, mapA gene, local chicken

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

    Science.gov (United States)

    Mavri, Ana; Smole Možina, Sonja

    2013-01-01

    The potential for adaptive resistance of Campylobacter jejuni and Campylobacter coli after step-wise exposure to increasing sub-inhibitory concentrations of five biocides as triclosan, benzalkonium chloride, cetylpyridinium chloride, chlorhexidine diacetate and trisodium phosphate, was investigated, to identify the mechanisms underlying resistance. The biocide resistance and cross-resistance to the antimicrobials erythromycin and ciprofloxacin, and to sodium dodecyl sulphate, were examined according to the broth microdilution method. The presence of active efflux was studied on the basis of restored sensitivity in the presence of the efflux pump inhibitors phenylalanine-arginine beta-naphthylamide, 1-(1-naphthylmethyl)-piperazine, cyanide 3-chlorophenylhydrazone, verapamil and reserpine. Changes in the outer membrane protein profiles and morphological changes in adapted strains were studied, as compared with the parent strains. Repeated exposure of C. jejuni and C. coli to biocides resulted in partial increases in tolerance to biocides itself, to other biocides and antimicrobial compounds. The developed resistance was stable for up to 10 passages in biocide-free medium. More than one type of active efflux was identified in adapted strains. These adapted strains showed different alterations to their outer membrane protein profiles, along with morphological changes. The data presented here suggest that different mechanisms are involved in adaptation to biocides and that this adaptation is unique to each strain of Campylobacter and does not result from a single species-specific mechanism. Copyright © 2012 Elsevier B.V. All rights reserved.

  16. Effects of short-chain nitrocompounds against Campylobacter jejuni and Campylobacter coli in vitro.

    Science.gov (United States)

    Horrocks, S M; Jung, Y S; Huwe, J K; Harvey, R B; Ricke, S C; Carstens, G E; Callaway, T R; Anderson, R C; Ramlachan, N; Nisbet, D J

    2007-03-01

    Effects of 2-nitro-1-propanol, 2-nitroethanol, nitroethane, and 2-nitro-methyl-propionate (0, 10, and 20 mM) on growth of Campylobacter jejuni were tested during culture in Bolton broth adjusted to pH 5.6, 7.0, or 8.2. The nitrocompounds were similarly tested against C. coli but at pH 8.2 only. Viable cell counts measured during incubation revealed main effects (P nitrocompounds on the survivability of C. jejuni. An effect of pH (P nitrocompounds was observed, with greater inhibition observed at pH 8.2 than at pH 5.6 or 7.0 for nitroethane, 2-nitro-l-propanol, and 2-nitroethanol, but not for 2-nitro-methyl-propionate, which showed greatest inhibition at pH 5.6. Except for 2-nitro-methyl-propionate, which was ineffective, all nitrocompounds elicited similar effects on C. coli. The effect of nitroethane and 2-nitro-l-propanol (10 mM) on naturally occurring Campylobacter was investigated during incubation of porcine fecal suspensions, where Campylobacter concentrations decreased more rapidly (P < 0.05) in suspensions with added 2-nitro-l-propanol than in unsupplemented or nitroethane-supplemented suspensions, thus reiterating the superior inhibitory effect of 2-nitro-l-propanol.

  17. Impaired Fitness and Transmission of Macrolide-Resistant Campylobacter jejuni in Its Natural Host

    Science.gov (United States)

    Luangtongkum, Taradon; Shen, Zhangqi; Seng, Virginia W.; Sahin, Orhan; Jeon, Byeonghwa; Liu, Peng

    2012-01-01

    Campylobacter jejuni is a major zoonotic pathogen transmitted to humans via the food chain and is prevalent in chickens, a natural reservoir for this pathogenic organism. Due to the importance of macrolide antibiotics in clinical therapy of human campylobacteriosis, development of macrolide resistance in Campylobacter has become a concern for public health. To facilitate the control of macrolide-resistant Campylobacter, it is necessary to understand if macrolide resistance affects the fitness and transmission of Campylobacter in its natural host. In this study we conducted pairwise competitions and comingling experiments in chickens using clonally related and isogenic C. jejuni strains, which are either susceptible or resistant to erythromycin (Ery). In every competition pair, Ery-resistant (Eryr) Campylobacter was consistently outcompeted by the Ery-susceptible (Erys) strain. In the comingling experiments, Eryr Campylobacter failed to transmit to chickens precolonized by Erys Campylobacter, while isogenic Erys Campylobacter was able to transmit to and establish dominance in chickens precolonized by Eryr Campylobacter. The fitness disadvantage was linked to the resistance-conferring mutations in the 23S rRNA. These findings clearly indicate that acquisition of macrolide resistance impairs the fitness and transmission of Campylobacter in chickens, suggesting that the prevalence of macrolide-resistant C. jejuni will likely decrease in the absence of antibiotic selection pressure. PMID:22183170

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

    DEFF Research Database (Denmark)

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

    2004-01-01

    A DNA microarray for detection of Campylobacter spp. was recently developed and applied to detect Campylobacter spp. directly from chicken feces. Sixty-five pooled chicken cloacal swab samples from 650 individual broiler chickens were included in the study. The results of Campylobacter sp....... detection obtained with DNA microarrays were compared to those obtained by conventional culture and gel electrophoresis. By conventional culture, 60% of the samples were positive for either Campylobacter jejuni or Campylobacter coli. By PCR and capillary electrophoresis, 95% of the samples were positive...... for Campylobacter spp., whereas with DNA microarrays all samples were positive for Campylobacter spp. By application of DNA microarray analysis, the isolates in 4 samples (6%) could not be identified to the species level, whereas by PCR-capillary electrophoresis, the isolates in 12 samples (19%) remained...

  19. Risk factors associated with Campylobacter jejuni infections in Curacao, Netherlands Antilles

    NARCIS (Netherlands)

    H.P. Endtz (Hubert); L. de Haan (Lidewij); R. van Koningsveld (Rinske); Y. Halabi; N.P.W.C.J. van den Braak (Nicole); B.I. Kesztyus; C.W. Ang (Wim); I. Gerstenbluth; E. Leyde; A. Ott (Alewijn); F.G. Rodgers; R.P.A.J. Verkooyen (Roel); D.L. Woodward; A.F. van Belkum (Alex); L.J. Price; H. West; P.C.R. Godschalk (Peggy)

    2003-01-01

    textabstractA steady increase in the incidence of Guillain-Barre syndrome (GBS) with a seasonal preponderance, almost exclusively related to Campylobacter jejuni, and a rise in the incidence of laboratory-confirmed Campylobacter enteritis have been reported from Curacao,

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

    OpenAIRE

    Whiley, Harriet; McLean, Ryan; Ross, Kirstin

    2016-01-01

    Worldwide, Campylobacter is a significant cause of gastrointestinal illness. It is predominately considered a foodborne pathogen, with human exposure via non-food transmission routes generally overlooked. Current literature has been exploring environmental reservoirs of campylobacteriosis including potential wildlife reservoirs. Given the close proximity between lizards and human habitats in Central Australia, this study examined the presence of Campylobacter jejuni from lizard faeces collect...

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

    DEFF Research Database (Denmark)

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

    2003-01-01

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

  2. Antibiotic Resistance and Prevalence of Campylobacter jejuni and Campylobacter coli in Poultry Liver

    Directory of Open Access Journals (Sweden)

    A. Saadatmand

    2017-10-01

    Full Text Available Background and Objective: Campylobacter is a common type of bacteria in humans and poultry, which generally accounts for various diseases in humans, such as gastroenteritis. The poultry digestive system contains a high level of these bacteria. The aim of this study was to evaluate the prevalence of C. jejuni and C. coli in the poultry liver packed for marketing and determine the antibiotic resistance of the isolates. Materials and Methods: This cross-sectional study was conducted in the spring of 2016 in the city of Hamadan, Iran. A total of 80 samples of packed chicken liver were collected from the stores supplying meat and poultry products in Hamadan. The enrichment of the liver samples was performed in brucella broth; subsequently, separation was carried out on Campylobacter selective agar. The presence of bacteria was confirmed by the implementation of chemical diagnostic tests and direct microscopic observation. Finally, the antibiotic resistance of the isolates was tested using disk diffusion method. Results: According to the results, Campylobacter had a prevalence rate of 90%, 73.61% and 26.39% of which were C. jejuni and C. coli, respectively. Out of the 12 antibiotic discs used in this study, the highest resistance (79% and sensitivity (99% rates were observed for cotrimoxazole (10 µg and gentamycin (10 µg, respectively. Conclusion: The packed poultry liver in Hamadan had a relatively high prevalence of C. jejuni and C. coli. Therefore, the consumers should be careful about the cooking time and using this food. Accordingly, they can prevent the dissemination of this bacteria by cooking the liver at a temperature of above 70°C for 20 min and properly washing the devices before cooking this product. Additionally, the elderly, children, and those with immunodeficiency are recommended to avoid eating poultry liver.

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

    Science.gov (United States)

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

    2016-01-01

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

  4. Impaired Fitness and Transmission of Macrolide-Resistant Campylobacter jejuni in Its Natural Host

    OpenAIRE

    Luangtongkum, Taradon; Shen, Zhangqi; Seng, Virginia W.; Sahin, Orhan; Jeon, Byeonghwa; Liu, Peng; Zhang, Qijing

    2012-01-01

    Campylobacter jejuni is a major zoonotic pathogen transmitted to humans via the food chain and is prevalent in chickens, a natural reservoir for this pathogenic organism. Due to the importance of macrolide antibiotics in clinical therapy of human campylobacteriosis, development of macrolide resistance in Campylobacter has become a concern for public health. To facilitate the control of macrolide-resistant Campylobacter, it is necessary to understand if macrolide resistance affects the fitness...

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

    DEFF Research Database (Denmark)

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

    2001-01-01

    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 were clearly...

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

    DEFF Research Database (Denmark)

    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...

  7. PFGE, Lior Serotype, and Antimicrobial Resistance Patterns Among Campylobacter jejuni Isolated from Travelers and US Military Personnel with Acute Diarrhea in Thailand, 1998-2003

    Science.gov (United States)

    2010-01-01

    gastroenteritis . Background Campylobacter jejuni is a major cause of gastroenteritis worldwide, especially in children, travelers, and military personnel...of gastroenteritis caused by Campylobacter jejuni. J Clin Microbial 2003, 41:4733-4739. 20. Beecham HJ, Lebron Cl. Echeverria P: Short report...and antimicrobial resistance patterns among Campylobacter jejuni isolated from travelers and US military personnel with acute diarrhea in Thailand

  8. Global Distribution of Campylobacter jejuni Penner Serotypes: A Systematic Review.

    Directory of Open Access Journals (Sweden)

    Brian L Pike

    Full Text Available Penner serotyping has been the principal method for differentiating Campylobacter isolates since its inception. Campylobacter capsule polysaccharide (CPS, the principal serodeterminant on which Penner serotyping is based, is presently of interest as a vaccine component. To determine the required valency of an effective CPS-based vaccine, a comprehensive understanding of CPS distribution is needed. Because of the association between Penner serotype and CPS, we conducted a systematic review to estimate the frequency and distribution of Penner serotypes associated with cases of Campylobacteriosis. In total, more than 21,000 sporadic cases of C. jejuni cases were identified for inclusion. While regional variation exists, distribution estimates indicate that eight serotypes accounted for more than half of all sporadic diarrheal cases globally and three serotypes (HS4 complex, HS2, and HS1/44 were dominant inter-regionally as well as globally. Furthermore, a total of 17 different serotypes reached a representation of 2% or greater in at least one of the five regions sampled. While this review is an important first step in defining CPS distribution, these results make it clear that significant gaps remain in our knowledge. Eliminating these gaps will be critical to future vaccine development efforts.

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

    Directory of Open Access Journals (Sweden)

    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.

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

    Directory of Open Access Journals (Sweden)

    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.

  11. Role of alkyl hydroperoxide reductase (AhpC in the biofilm formation of Campylobacter jejuni.

    Directory of Open Access Journals (Sweden)

    Euna Oh

    Full Text Available Biofilm formation of Campylobacter jejuni, a major cause of human gastroenteritis, contributes to the survival of this pathogenic bacterium in different environmental niches; however, molecular mechanisms for its biofilm formation have not been fully understood yet. In this study, the role of oxidative stress resistance in biofilm formation was investigated using mutants defective in catalase (KatA, superoxide dismutase (SodB, and alkyl hydroperoxide reductase (AhpC. Biofilm formation was substantially increased in an ahpC mutant compared to the wild type, and katA and sodB mutants. In contrast to the augmented biofilm formation of the ahpC mutant, a strain overexpressing ahpC exhibited reduced biofilm formation. A perR mutant and a CosR-overexpression strain, both of which upregulate ahpC, also displayed decreased biofilms. However, the introduction of the ahpC mutation to the perR mutant and the CosR-overexpression strain substantially enhanced biofilm formation. The ahpC mutant accumulated more total reactive oxygen species and lipid hydroperoxides than the wild type, and the treatment of the ahpC mutant with antioxidants reduced biofilm formation to the wild-type level. Confocal microscopy analysis showed more microcolonies were developed in the ahpC mutant than the wild type. These results successfully demonstrate that AhpC plays an important role in the biofilm formation of C. jejuni.

  12. Significance of phage-host interactions for biocontrol of Campylobacter jejuni in food

    DEFF Research Database (Denmark)

    Athina, Zampara; Sørensen, Martine Camilla Holst; Elsser-Gravesen, Anne

    2017-01-01

    Poultry meat is the main source of Campylobacter jejuni foodborne disease. Currently, no effective control measures prevent C. jejuni from contaminating poultry meat. However, post-harvest phage treatment is a promising biocontrol strategy that has not yet been explored. Here we identified phages...... most effective phages (F356 showing 0.49 and F357 showing 0.55 log reductions, respectively) led to a 0.73 log reduction of C. jejuni on artificially contaminated chicken skin. Our study shows that poly-phage treatment at 5 °C can be more effective against C. jejuni compared to single phage application...

  13. [The relationship of Campylobacter jejuni subsp. jejuni enterotoxigenicity and the increase of cAMP and electrolyte changes in the rat intestine].

    Science.gov (United States)

    Fernández, H; Toro, J

    1998-08-01

    Small intestine alterations produced by the enterotoxigenic capacity of Campylobacter jejuni subsp. jejuni are similar to the hydric, electrolytic and pathological changes caused by choleraic and thermolabile Escherichia coli toxins. To study the enterotoxigenic capacity of 4 strains of Campylobacter jejuni subsp. jejuni using the intestinal loop model. Rat intestinal loops were inoculated with culture filtrates of the four strains. Enterotoxigenicity was assessed by fluid accumulation, the increase in Na+ and Cl- in the loop fluid, and cAMP increase in loop tissues. An enterotoxigenic Escherichia coil strain and sterile Brucella both were used as positive and negative controls, respectively. The filtrates of two strains produced fluid accumulation in the loops, significantly increased Na+ and Cl- secretion to the intestinal lumen and increased tissue cAMP levels. Some strains of Campylobacter jejuni subsp. jejuni are able to show enterotoxigenicity in vivo, increasing cAMP levels in the intestinal cells and altering electrolyte exchange mechanisms.

  14. Prevalence of Campylobacter jejuni, C.lari and C.coli in different ecological guilds and taxa of migrating birds

    NARCIS (Netherlands)

    Waldenstrom, J.; Broman, T.; Carlsson, I.; Hasselquist, D.; Achterberg, R.P.; Wagenaar, J.A.; Olsen, B.

    2002-01-01

    A total of 1,794 migrating birds trapped at a coastal site in southern Sweden were sampled for detection of Campylobacter spp. All isolates phenotypically identified as Campylobacter jejuni and a subset of those identified as non-C. jejuni were identified to the species level by PCR-based

  15. Quantification and differentiation of Campylobacter jejuni and Campylobacter coli in raw chicken meats using a real-time PCR method.

    Science.gov (United States)

    Hong, Joonbae; Jung, Woo Kyung; Kim, Jun Man; Kim, So Hyun; Koo, Hye Cheong; Ser, Junghee; Park, Yong Ho

    2007-09-01

    Campylobacter species are one of the most common causes of bacterial diarrhea in humans worldwide. The consumption of foods contaminated with two Campylobacter species, C. jejuni and C. coli, is usually associated with most of the infections in humans. In this study, a rapid, reliable, and sensitive multiplex real-time quantitative PCR was developed for the simultaneous detection, identification, and quantification of C. jejuni and C. coli. In addition, the developed method was applied to the 50 samples of raw chicken meat collected from retail stores in Korea. C. jejuni and C. coli were detected in 88 and 86% of the samples by real-time quantitative PCR and the conventional microbiological method, respectively. The specificity of the primer and probe sets was confirmed with 30 C. jejuni, 20 C. coli, and 35 strains of other microbial species. C. jejuni and C. coli could be detected with high specificity in less than 4 h, with a detection limit of 1 log CFU/ml by the developed real-time PCR. The average counts (log CFU per milliliter) of C. jejuni or C. coli obtained by the conventional methods and by the real-time PCR assay were statistically correlated with a correlation coefficient (R2) between 0.73 and 0.78. The real-time PCR assay developed in this study is useful for screening for the presence and simultaneous differential quantification of C. jejuni and C. coli.

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

    Directory of Open Access Journals (Sweden)

    Reza Shahrokhabadi

    2013-11-01

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

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

    Science.gov (United States)

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

    2015-01-01

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

  18. Variation in Campylobacter jejuni culturability in presence of Acanthamoeba castellanii Neff.

    Science.gov (United States)

    Reyes-Batlle, María; Girbau, Cecilia; López-Arencibia, Atteneri; Sifaoui, Ines; Liendo, Aitor Rizo; Bethencourt Estrella, Carlos J; García Méndez, Ana B; Chiboub, Olfa; Hajaji, Soumaya; Fernández-Astorga, Aurora; Valladares, Basilio; Martínez-Carretero, Enrique; Piñero, José E; Lorenzo-Morales, Jacob

    2017-12-01

    Free-living amoebae (FLA) are protozoa that are widely distributed in the environment mainly in water and soil related habitats. These amoebae have also been reported to be associated with some bacterial pathogens for humans such as Campylobacter spp. The species C. jejuni is the causative agent of about 90% of human campylobacteriosis cases worldwide and this disease may even end up in severe autoimmune sequelae as Guillain-Barré syndrome (GBS). In this study, the interactions between the strain Acanthamoeba castellanii Neff and Campylobacter jejuni was investigated. Campylobacter jejuni was coincubated with Acanthamoeba castellanii Neff trophozoites at different temperatures, in order to evaluate the C. jejuni ability to grow in presence A. castellanii culture and Acanthamoeba Conditioned Medium (ACM). C. jejuni was coincubated with A. castellanii axenic culture at different temperatures in aerobic conditions. Our results revealed that bacteria were still cultivable (Blood Agar medium, at 37 °C, in microaerophilic atmosphere) after a 14 days C. jejuni - A. castellanii coculture, comparing with C. jejuni alone, which was only cultivable for 24 h. Copyright © 2017 Elsevier Inc. All rights reserved.

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

    Science.gov (United States)

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

    2016-09-01

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

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

    Science.gov (United States)

    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...

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

    Directory of Open Access Journals (Sweden)

    Heath Sekou

    2007-02-01

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

  2. A Real-Time PCR Assay for the Detection of Campylobacter jejuni in Foods after Enrichment Culture

    OpenAIRE

    Sails, Andrew D.; Fox, Andrew J.; Bolton, Frederick J.; Wareing, David R. A.; Greenway, David L. A.

    2003-01-01

    A real-time PCR assay was developed for the quantitative detection of Campylobacter jejuni in foods after enrichment culture. The specificity of the assay for C. jejuni was demonstrated with a diverse range of Campylobacter species, related organisms, and unrelated genera. The assay had a linear range of quantification over six orders of magnitude, and the limit of detection was approximately 12 genome equivalents. The assay was used to detect C. jejuni in both naturally and artificially cont...

  3. Peroxynitrite scavenging by Campylobacter jejuni truncated hemoglobin P.

    Science.gov (United States)

    Ascenzi, Paolo; Pesce, Alessandra

    2017-12-01

    Truncated hemoglobins (trHb) are present in protozoa, cyanobacteria, nemertean, bacteria, algae, and plants. They are characterized by the 2-on-2 topology and are ordered in four phylogenetic groups (I or N, II or O, III or P, and IV or Q). Several functions have been attributed to trHbs including the inactivation of reactive nitrogen and oxygen species, permitting the survival of microorganisms in the host. Here, the kinetics of peroxynitrite scavenging by ferric Campylobacter jejuni truncated hemoglobin P [i.e., Cj-trHbP(III)] in the absence and presence of CO 2 , between pH 6.3 and 7.9, and 25.0 °C, is reported. Mixing the Cj-trHbP(III) solution with peroxynitrite solution brings about absorption spectral changes at 302 nm reflecting the disappearance of this endogenous toxicant and cytotoxic effector against pathogens. CO 2 affects only the rate of spontaneous decay of peroxynitrite without affecting the scavenging activity of Cj-trHbP(III). Moreover, the Cj-trHbP(III)-mediated isomerization of peroxynitrite is facilitated at low pH, indicating that peroxynitrous acid is the reactive species. The high reactivity of Cj-trHbP(III) towards peroxynitrite has been ascribed to the peroxidase-like geometry of the metal center. To investigate the protective role of Cj-trHbP(III) against peroxynitrite-mediated nitration, the relative yield of nitro-L-tyrosine formed by the reaction of peroxynitrite with free L-tyrosine was determined. According to fast kinetics of peroxynitrite isomerization by Cj-trHbP(III), this 2-on-2 globin impairs L-tyrosine nitrosylation. Present data suggest that Cj-trHbP could help the survival of C. jejuni.

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

    Directory of Open Access Journals (Sweden)

    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.

  5. A Quantitative Real-Time PCR Approach for Assessing Campylobacter jejuni and Campylobacter coli Colonization in Broiler Herds.

    Science.gov (United States)

    Haas, Katrin; Overesch, Gudrun; Kuhnert, Peter

    2017-04-01

    Human campylobacteriosis is a major public health concern in developed countries, with Campylobacter jejuni and Campylobacter coli from poultry recognized as the main source of human infection. Identification of Campylobacter-positive broiler herds before slaughter is essential for implementing measures to avoid carryover of pathogens via the slaughter process into the food chain. However, appropriate methods that have been validated for testing poultry flocks antemortem are lacking for Campylobacter. A quantitative real-time PCR (qPCR) that allows simultaneous detection and quantification of C. jejuni and C. coli was adapted and optimized to be applied on boot socks. The adjusted qPCR serves as an easy, sensitive, and quantitative method for Campylobacter detection in poultry flocks antemortem by analysis of boot socks. An adequate correlation was found between qPCR and culture, as well as between boot socks and cecal samples, which are regarded as the "gold standard." Therefore, boot sock sampling followed by qPCR analysis provides a reliable and simple method for assessing Campylobacter load within a flock prior to slaughter. The approach allows categorization of broiler herds into negative, low, moderate, or high Campylobacter colonization. Based on the results of this new approach, risk assessment models, such as evaluating the possible effect of sorting flocks before slaughter, can be easily implemented. Similarly, targeted identification of highly colonized flocks for improvement of biosecurity measures at the farm level will become feasible, presenting an opportunity to increase food safety.

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

    Science.gov (United States)

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

    2016-10-01

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

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

    Directory of Open Access Journals (Sweden)

    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.

  8. Isolation and polymerase chain reaction-based detection of Campylobacter jejuni and Campylobacter coli from poultry in the Philippines.

    Science.gov (United States)

    Magistrado, P A; Garcia, M M; Raymundo, A K

    2001-10-22

    The polymerase chain reaction (PCR) and the conventional culture method of detecting thermophilic Campylobacter species in duck and chicken samples from two locations in the province of Laguna, Philippines, were compared. Three Campylobacter jejuni and five C. coli strains were isolated from a total of 135 duck and chicken samples from both methods. The PCR technique, however, was found to be more sensitive, accurate and rapid than the conventional culture method. The specificity of two sets of published primers, C442-C490 (specific for C. jejuni, C. coli and C. lari) and CL2-CR3 (specific for C. jejuni) were confirmed with reference and field strains. To improve detection, a lysate was prepared by boiling cells in Triton X-100, and then used as template for PCR to detect Campylobacter from spiked and naturally contaminated chicken rinse. For spiked chicken samples, a 17-h Meuller-Hinton Broth enrichment for the chicken rinse resulted in an improved sensitivity at 31.7 CFU/g using C442-C490. This enrichment-PCR tandem also detected thermophilic Campylobacter from 1 out of 21 native chicken samples from a wet market. To our knowledge, this is the first report of thermophilic Campylobacter isolation from poultry in the Philippines. The approaches described here could serve as a basis for future surveillance and/or epidemiological studies on this emerging foodborne pathogen.

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

    OpenAIRE

    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 ...

  10. Campylobacter jejuni prevalence and hygienic quality of retail bovine ground meat in Finland.

    Science.gov (United States)

    Llarena, A-K; Sivonen, K; Hänninen, M-L

    2014-05-01

    Detection of common genotypes of Campylobacter jejuni among Finnish human and bovine isolates, suggested that bovines may be a source for zoonotic Camp. jejuni infection. In addition, a Finnish epidemiological study implied the tasting and eating raw or undercooked beef as risk factors for acquiring campylobacteriosis. We therefore performed a study on the occurrence of Camp. jejuni in retail bovine ground meat in Helsinki by the use of both cultivation and PCR. During 2011 and 2012, 175 bovine ground meat samples were collected. None of the samples were Campylobacter positive by cultivation, and only one sample (0.6%) was Camp. jejuni positive by the use of PCR on template extracted directly from ground meat. According to our findings, Finnish bovine ground meat is an unlikely source for human campylobacteriosis. Additionally, the hygienic quality of bovine ground meat at retail level was screened and found to be good when monitored by aerobic micro-organisms, total thermotolerant coliforms and Eshericha coli. This study provides the first data on the occurrence of the zoonotic pathogen Campylobacter jejuni in Finnish bovine ground meat. This knowledge is important as part of future Campylobacter risk assessment, management and monitoring programs, particularly when assessing the relative attribution of poultry, pork and bovine meat to the burden of human campylobacteriosis. According to our results, Finnish bovine ground meat at retail level is of good hygienic quality. © 2013 The Society for Applied Microbiology.

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

    NARCIS (Netherlands)

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

    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%),

  12. Campylobacter.

    Science.gov (United States)

    Fitzgerald, Collette

    2015-06-01

    Campylobacter continues to be one of the most common bacterial causes of diarrheal illness in the United States and worldwide. Infection with Campylobacter causes a spectrum of diseases including acute enteritis, extraintestinal infections, and postinfectious complications. The most common species of Campylobacter associated with human illness is Campylobacter jejuni, but other Campylobacter species can also cause human infections. This comprehensive review includes discussion of the taxonomy, clinical manifestations of infection, epidemiology and the different methods of laboratory detection of Campylobacter. Published by Elsevier Inc.

  13. Campylobacter jejuni in the red squirrel (Sciurus vulgaris) population of Southern Italy.

    Science.gov (United States)

    Dipineto, Ludovico; Gargiulo, Antonio; Cuomo, Alessandra; Santaniello, Antonio; Sensale, Mariangela; Borrelli, Luca; D'Angelo, Livia; Menna, Lucia F; Fioretti, Alessandro

    2009-01-01

    Rectal swab samples were collected from 60 red squirrels (Sciurus vulgaris) from July 2006 to April 2007 in Southern Italy. Samples were tested for Campylobacter jejuni and C. coli by culture methods and suspected colonies were then confirmed by polymerase chain reaction. C. jejuni was detected in 5/60 (8.3%) samples examined but infection status was not related to age or sex and C. coli was not isolated. This is believed to be the first report of C. jejuni infection in the red squirrel.

  14. Proteins involved in Campylobacter jejuni 81-176 recovery after high-pressure treatment.

    Science.gov (United States)

    Bièche, Clémence; de Lamballerie, Marie; Federighi, Michel; Le Bail, Alain; Tresse, Odile

    2010-02-01

    Campylobacteriosis has been recognized as the major bacterial food-borne infection worldwide. Campylobacter, especially C. jejuni, contaminate mainly poultry meat. Although more sensitive than other food-borne pathogens to many stresses, C. jejuni can survive food processing and go on to reach its final reservoir (the human gut). Genomic analyses of this organism indicate a lack of genes described in other gram-negative bacteria to overcome stresses. The high-pressure recovery response of C. jejuni 81-176 was analyzed from two-dimensional electrophoretic profiles of the cytoplasmic proteome. The main cellular mechanisms controlling the down- and upregulated proteins are discussed.

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

    Directory of Open Access Journals (Sweden)

    Tadhg eÓ Cróinín

    2012-03-01

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

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

    DEFF Research Database (Denmark)

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

    2006-01-01

    of resistance among C. jejuni were observed for tetracycline, nalidixic acid and ciprofloxacin, whereas macrolide resistance was rarely detected. C. jejuni originating from other poultry meat (mainly duck and turkey meat) exhibited the highest occurrences of antimicrobial resistance monitored; approximately one....... 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.......Campylobacter jejuni isolated from raw poultry meat collected at retail shops in Denmark in the period 1996-2003 were tested for susceptibility to seven antimicrobial agents. The food samples consisted of raw chicken meat and other raw poultry meat of domestic or imported origin. The highest levels...

  17. High hydrostatic pressure resistance of Campylobacter jejuni after different sublethal stresses.

    Science.gov (United States)

    Sagarzazu, N; Cebrián, G; Condón, S; Mackey, B; Mañas, P

    2010-07-01

    To study the development of resistance responses in Campylobacter jejuni to high hydrostatic pressure (HHP) treatments after the exposure to different stressful conditions that may be encountered in food-processing environments, such as acid pH, elevated temperatures and cold storage. Campylobacter jejuni cells in exponential and stationary growth phase were exposed to different sublethal stresses (acid, heat and cold shocks) prior to evaluate the development of resistance responses to HHP. For exponential-phase cells, neither of the conditions tested increased nor decreased HHP resistance of C. jejuni. For stationary-phase cells, acid and heat adaptation-sensitized C. jejuni cells to the subsequent pressure treatment. On the contrary, cold-adapted stationary-phase cells developed resistance to HHP. Whereas C. jejuni can be classified as a stress sensitive micro-organism, our findings have demonstrated that it can develop resistance responses under different stressing conditions. The resistance of stationary phase C. jejuni to HHP was increased after cells were exposed to cold temperatures. The results of this study contribute to a better knowledge of the physiology of C. jejuni and its survival to food preservation agents. Results here presented may help in the design of combined processes for food preservation based on HHP technology. © 2009 The Authors. Journal compilation © 2009 The Society for Applied Microbiology.

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

    Directory of Open Access Journals (Sweden)

    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.

  19. Virulence and genomic feature of multidrug resistant Campylobacter jejuni isolated from broiler chicken

    Directory of Open Access Journals (Sweden)

    Haihong Hao

    2016-10-01

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

  20. Antimicrobial susceptibility profiles of human Campylobacter jejuni isolates and association with phylogenetic lineages

    Directory of Open Access Journals (Sweden)

    Wonhee eCha

    2016-04-01

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

  1. [Evaluation of usefulness of commercial recomwell Campylobacter enzyme--linked immunosorbent assays for routine serodiagnosis of Campylobacter jejuni and Campylobacter coli infections].

    Science.gov (United States)

    Rokosz, Natalia; Rastawicki, Waldemar; Jagielski, Marek

    2008-01-01

    The commercially available enzyme-linked immunosorbent assays (ELISA recomWell Campylobacter) from Mikrogen was evaluated for the diagnosis of Campylobacter jejuni and Campylobacter coli infections. Serum samples from 20 healthy controls, 44 persons with symptoms of primary Campylobacter infection and 24 serum samples from patients with Yersinia enterocolitica or Salmonella infections were tested. This ELISA assay detects IgA and IgG antibodies against three recombinant antigens of the Campylobacter jejuni and Campylobacter coli: OMP 18 (18 kDa), PEB4 (31 kDa) and P39 (39 kDa). The healthy controls showed significantly lower antibody titers in all two immunoglobulin classes. The IgA antibodies were diagnosed only in 2 (18.2%) serum samples obtained from patients with bacteriologically confirmed campylobacteriosis. The presence of IgG antibodies was confirmed in 82% of serum samples. Furthermore, we showed that 66.7% of the 33 serum samples obtained from the patients suspected for campylobacteriosis not confirmed by isolation, were positive for IgG and 15.2% for IgA antibodies. We observed also not specific reactions in ELISA recom Well Campylobacter with sera obtained form patients with yersiniosis and salmonelosis. This study demonstrates the usefulness of commercially available assay for the routine diagnosis of Campylobacter infection but with some limitations.

  2. Key role of Mfd in the development of fluoroquinolone resistance in Campylobacter jejuni.

    Directory of Open Access Journals (Sweden)

    Jing Han

    2008-06-01

    Full Text Available Campylobacter jejuni is a major food-borne pathogen and a common causative agent of human enterocolitis. Fluoroquinolones are a key class of antibiotics prescribed for clinical treatment of enteric infections including campylobacteriosis, but fluoroquinolone-resistant Campylobacter readily emerges under the antibiotic selection pressure. To understand the mechanisms involved in the development of fluoroquinolone-resistant Campylobacter, we compared the gene expression profiles of C. jejuni in the presence and absence of ciprofloxacin using DNA microarray. Our analysis revealed that multiple genes showed significant changes in expression in the presence of a suprainhibitory concentration of ciprofloxacin. Most importantly, ciprofloxacin induced the expression of mfd, which encodes a transcription-repair coupling factor involved in strand-specific DNA repair. Mutation of the mfd gene resulted in an approximately 100-fold reduction in the rate of spontaneous mutation to ciprofloxacin resistance, while overexpression of mfd elevated the mutation frequency. In addition, loss of mfd in C. jejuni significantly reduced the development of fluoroquinolone-resistant Campylobacter in culture media or chickens treated with fluoroquinolones. These findings indicate that Mfd is important for the development of fluoroquinolone resistance in Campylobacter, reveal a previously unrecognized function of Mfd in promoting mutation frequencies, and identify a potential molecular target for reducing the emergence of fluoroquinolone-resistant Campylobacter.

  3. Rapid and Specific Methods to Differentiate Foodborne Pathogens, Campylobacter jejuni, Campylobacter coli, and the New Species Causing Spotty Liver Disease in Chickens, Campylobacter hepaticus.

    Science.gov (United States)

    Van, Thi Thu Hao; Anwar, Arif; Scott, Peter C; Moore, Robert J

    2018-01-22

    Campylobacter jejuni and Campylobacter coli play a major role in bacteria-related foodborne illness in humans. Recently, a newly identified species, Campylobacter hepaticus, was shown to be the causative agent of spotty liver disease in chickens. The pathogenic potential of C. hepaticus in humans is unknown. This new species contains genes usually used to detect C. jejuni and C. coli in DNA-based detection methods, such as the hippuricase (hipO) gene and the glyA (serine hydroxymethyltransferase) gene, with a high degree of similarity. Therefore, polymerase chain reaction (PCR) primers used to detect these species need to be evaluated carefully to prevent misidentification of these important Campylobacter species. A multiplex PCR was developed and optimized to simultaneously and specifically identify the presence of C. jejuni, C. coli, and C. hepaticus in chicken samples containing high-complexity microbiota. The assay represents a new diagnostic tool for investigating the epidemiology of Campylobacter colonization in poultry and environmental samples. It may also be applicable to the investigation of Campylobacter contamination in food and in outbreaks of campylobacteriosis.

  4. Identification of a functional type VI secretion system in Campylobacter jejuni conferring capsule polysaccharide sensitive cytotoxicity.

    Directory of Open Access Journals (Sweden)

    Nancy M C Bleumink-Pluym

    Full Text Available The pathogen Campylobacter jejuni is the principal cause of bacterial food-borne infections. The mechanism(s that contribute to bacterial survival and disease are still poorly understood. In other bacterial species, type VI secretion systems (T6SS are increasingly recognized to contribute to bacterial pathogenesis by toxic effects on host cells or competing bacterial species. Here we report the presence of a functional Type VI secretion system in C. jejuni. Proteome and genetic analyses revealed that C. jejuni strain 108 contains a 17-kb T6SS gene cluster consisting of 13 T6SS-conserved genes, including the T6SS hallmark genes hcp and vgrG. The cluster lacks an ortholog of the ClpV ATPase considered important for T6SS function. The sequence and organization of the C. jejuni T6SS genes resemble those of the T6SS located on the HHGI1 pathogenicity island of Helicobacter hepaticus. The C. jejuni T6SS is integrated into the earlier acquired Campylobacter integrated element CJIE3 and is present in about 10% of C. jejuni isolates including several isolates derived from patients with the rare clinical feature of C. jejuni bacteremia. Targeted mutagenesis of C. jejuni T6SS genes revealed T6SS-dependent secretion of the Hcp needle protein into the culture supernatant. Infection assays provided evidence that the C. jejuni T6SS confers contact-dependent cytotoxicity towards red blood cells but not macrophages. This trait was observed only in a capsule-deficient bacterial phenotype. The unique C. jejuni T6SS phenotype of capsule-sensitive contact-mediated hemolysis represents a novel evolutionary pathway of T6SS in bacteria and expands the repertoire of virulence properties associated with T6SS.

  5. Identification of a functional type VI secretion system in Campylobacter jejuni conferring capsule polysaccharide sensitive cytotoxicity.

    Science.gov (United States)

    Bleumink-Pluym, Nancy M C; van Alphen, Lieke B; Bouwman, Lieneke I; Wösten, Marc M S M; van Putten, Jos P M

    2013-01-01

    The pathogen Campylobacter jejuni is the principal cause of bacterial food-borne infections. The mechanism(s) that contribute to bacterial survival and disease are still poorly understood. In other bacterial species, type VI secretion systems (T6SS) are increasingly recognized to contribute to bacterial pathogenesis by toxic effects on host cells or competing bacterial species. Here we report the presence of a functional Type VI secretion system in C. jejuni. Proteome and genetic analyses revealed that C. jejuni strain 108 contains a 17-kb T6SS gene cluster consisting of 13 T6SS-conserved genes, including the T6SS hallmark genes hcp and vgrG. The cluster lacks an ortholog of the ClpV ATPase considered important for T6SS function. The sequence and organization of the C. jejuni T6SS genes resemble those of the T6SS located on the HHGI1 pathogenicity island of Helicobacter hepaticus. The C. jejuni T6SS is integrated into the earlier acquired Campylobacter integrated element CJIE3 and is present in about 10% of C. jejuni isolates including several isolates derived from patients with the rare clinical feature of C. jejuni bacteremia. Targeted mutagenesis of C. jejuni T6SS genes revealed T6SS-dependent secretion of the Hcp needle protein into the culture supernatant. Infection assays provided evidence that the C. jejuni T6SS confers contact-dependent cytotoxicity towards red blood cells but not macrophages. This trait was observed only in a capsule-deficient bacterial phenotype. The unique C. jejuni T6SS phenotype of capsule-sensitive contact-mediated hemolysis represents a novel evolutionary pathway of T6SS in bacteria and expands the repertoire of virulence properties associated with T6SS.

  6. Prevalence and Characterization of Campylobacter jejuni Isolated from Retail Chicken in Tianjin, China.

    Science.gov (United States)

    Ma, Hui; Su, Yulan; Ma, Luyao; Ma, Lina; Li, Ping; Du, Xinjun; Gölz, Greta; Wang, Shuo; Lu, Xiaonan

    2017-06-01

    Campylobacter jejuni is an important foodborne pathogen worldwide; however, there is a lack of information on the prevalence and antibiotic-resistant profile of C. jejuni in the People's Republic of China. We determined the prevalence and characteristics of C. jejuni on the retail level in Tianjin, one of the five national central cities in China. A total of 227 samples of chicken wings, legs, and breasts were collected from supermarkets and wet markets; 42 of these samples were confirmed to be positive for Campylobacter contamination. The contamination rates of C. jejuni and other Campylobacter species were 13.7% (31 of 227 samples) and 5.7% (13 of 227 samples), respectively. A group of 31 C. jejuni isolates was subjected to antimicrobial susceptibility testing. All (100%) the selected isolates were resistant to ciprofloxacin and nalidixic acid; 77.4% were resistant to tetracycline, 67.7% to doxycycline, 35.5% to gentamicin, 25.8% to clindamycin and florfenicol, 19.4% to chloramphenicol, and 12.9% to erythromycin and azithromycin. A remarkably high proportion (41.9%) of multidrug-resistant isolates was identified. Multilocus sequence typing was conducted to study the population structure of the C. jejuni strains and their relationship to human isolates. The correlation between antimicrobial resistance traits and certain sequence types (STs) or clonal complexes was determined as well. A great genetic diversity of poultry isolates was identified, with 11 STs belonging to 6 clonal complexes and 11 singleton STs. The novel STs accounted for 40.9% (n = 9) of the 22 STs. ST-21, ST-353, ST-354, ST-443, ST-607, and ST-828 complexes had been previously identified from human isolates. This study revealed an extensive level of antimicrobial resistance and genetic diversity in C. jejuni isolated from chicken products in Tianjin, highlighting the necessity of performing enforced interventions to reduce Campylobacter prevalence in China.

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

    Science.gov (United States)

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

    2014-03-01

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

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

    Science.gov (United States)

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

    2017-01-01

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

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

    Directory of Open Access Journals (Sweden)

    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

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

    Science.gov (United States)

    Tangkham, Wannee; Janes, Marlene; LeMieux, Frederick

    2016-01-01

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

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

    Science.gov (United States)

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

  12. Het Guillain-Barré-syndroom na een Campylobacter jejuni-enteritis

    NARCIS (Netherlands)

    Beenen, L. F.; Scholten, H. G.

    1990-01-01

    A 7-year-old boy nine days before onset of a Guillain-Barré-syndrome had had enteritis caused by Campylobacter jejuni. The clinical signs were restricted to minor motor and sensory deficits in the limbs; 5 weeks after the onset of the syndrome, recovery was complete. So far, 16 cases of

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

    DEFF Research Database (Denmark)

    Smigic, Nada; Rajkovic, Andreja; Arneborg, Nils

    2011-01-01

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

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

    Science.gov (United States)

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

  15. Identification of a functional type VI secretion system in Campylobacter jejuni conferring capsule polysaccharide sensitive cytotoxicity

    NARCIS (Netherlands)

    Bleumink-Pluym, Nancy M C; van Alphen, Lieke B; Bouwman, Lieneke I; Wösten, MM; van Putten, Jos P M

    2013-01-01

    The pathogen Campylobacter jejuni is the principal cause of bacterial food-borne infections. The mechanism(s) that contribute to bacterial survival and disease are still poorly understood. In other bacterial species, type VI secretion systems (T6SS) are increasingly recognized to contribute to

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

    NARCIS (Netherlands)

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

    2014-01-01

    Campylobacter jejuni is the most important cause of antecedent infections leading to Guillain-Barré syndrome (GBS) and Miller Fisher syndrome (MFS). The objective of the present study was to define the genetic diversity, population structure, and potential role of poultry in the transmission of

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

    NARCIS (Netherlands)

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

    2014-01-01

    Campylobacter jejuni is the most important cause of antecedent infections leading to Guillain-Barr, syndrome (GBS) and Miller Fisher syndrome (MFS). The objective of the present study was to define the genetic diversity, population structure, and potential role of poultry in the transmission of

  18. Ganglioside mimicry of Campylobacter jejuni lipopolysaccharides determines antiganglioside specificity in rabbits

    NARCIS (Netherlands)

    C.W. Ang (Wim); P.G. Noordzij (Peter); M.A. de Klerk; H.P. Endtz (Hubert); P.A. van Doorn (Pieter); J.D. Laman (Jon)

    2002-01-01

    textabstractThe core oligosaccharides of Campylobacter jejuni lipopolysaccharides (LPS) display molecular mimicry with gangliosides. Cross-reactive anti-LPS-antiganglioside antibodies have been implicated to show a crucial role in the pathogenesis of the Guillain-Barre and Miller

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

    Science.gov (United States)

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

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

    Science.gov (United States)

    Garofolo, Giuliano; Di Donato, Guido; Cianciavicchia, Silvia; Alessiani, Alessandra

    2015-01-01

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

  1. Distribution of Campylobacter jejuni capsular types, 2007-2012, Philadelphia, PA.

    Science.gov (United States)

    Garrigan, Charles; Ettela, Abora; Poly, Frédéric; Guerry, Patricia; Nachamkin, Irving

    2014-11-01

    The distribution of Campylobacter jejuni capsular serotypes in the Philadelphia region from 2007 to 2012 was determined using molecular methods. Compared with the last U.S. survey in 1990, there does not appear to be a major shift in circulating capsular types. Copyright © 2014 Elsevier Inc. All rights reserved.

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

    DEFF Research Database (Denmark)

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

    2017-01-01

    Here, we describe the methods for isolation, purification, and propagation of Campylobacter jejuni bacteriophages from samples expected to contain high number of phages such as chicken feces. The overall steps are (1) liberation of phages from the sample material; (2) observation of plaque-formin...

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

    Directory of Open Access Journals (Sweden)

    Francesca Marotta

    2015-01-01

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

  4. Consumer food preparation and its implication for survival of Campylobacter jejuni on chicken

    NARCIS (Netherlands)

    Bergsma, N.J.; Fischer, A.R.H.; Asselt, van E.D.; Zwietering, M.H.; Jong, de A.E.I.

    2007-01-01

    Purpose ¿ The disease burden caused by Campylobacter jejuni may be decreased by reduced consumption of undercooked chicken meat. However, little is known about consumer preparation of poultry and the effects of commonly applied cooking times on bacterial inactivation. This study aimed to answer

  5. Molecular evidence for dissemination of unique Campylobacter jejuni clones in Curacao, Netherlands Antilles

    NARCIS (Netherlands)

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

    2003-01-01

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

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

    NARCIS (Netherlands)

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

    2003-01-01

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

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

    Science.gov (United States)

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

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

    Science.gov (United States)

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

    2015-01-01

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

  9. Nachweis und Differenzierung von Campylobacter jejuni, C. coli und C. lari in Lebensmitteln mittels Quadruplex real-time PCR

    OpenAIRE

    Mayr, Anja Maria

    2009-01-01

    Es wurde ein Quadruplex real-time PCR-Verfahren zum parallelen Nachweis und zur Differenzierung der lebensmittelrelevanten thermophilen Campylobacter spp. Campylobacter jejuni, Campylobacter coli und Campylobacter lari in Geflügel, Fleisch, Fisch und Muscheln etabliert. Das Nachweissystem wurde hinsichtlich seiner Selektivität, Sensitivität, Effizienz, Präzision und Robustheit überprüft und im Vergleich zur konventionellen Diagnostik (kultureller Nachweis) anhand von 464 Feldproben validiert....

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

    Directory of Open Access Journals (Sweden)

    Sophie Kittler

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

  11. Generation and characterization of a novel recombinant scFv antibody specific for Campylobacter jejuni.

    Science.gov (United States)

    Nzuma, Ruramayi M; Liu, Fuquan; Grant, Irene R

    2018-04-07

    Campylobacter jejuni is a leading cause of foodborne illness worldwide, mainly due to consumption and handling of contaminated raw chicken. Rapid detection methods for C. jejuni are vital for monitoring contamination levels in chicken products and reducing human Campylobacteriosis cases. The 'gold standard' culture-based method of Campylobacter detection takes 3-5 days and is too slow to permit effective intervention. Immuno-based methods are faster, but usually necessitate use of animals or hybridoma technology to produce antibodies; making them difficult and expensive to produce. Here, we report the generation and characterization of recombinant single-chain variable fragment (scFv) antibodies specific for C. jejuni cells, and evaluation of one scFv antibody for an immunomagnetic separation-quantitative PCR (IMS-qPCR) method to rapidly, sensitively, and specifically detect low numbers of C. jejuni. An scFv antibody phage-display library was constructed using spleen mRNA derived from a rabbit immunized with gamma-irradiated C. jejuni cells. This library was screened by surface biopanning against C. jejuni whole cells. Enriched clones were analyzed by enzyme-linked immunosorbent assay (ELISA). Two scFv antibodies that strongly and specifically recognized C. jejuni cell were expressed in Escherichia coli. Western blot analysis showed that one antibody, scFv80, was expressed as a soluble protein and retained its specific and strong binding to C. jejuni cells. This recombinant monoclonal scFv antibody was purified and used to covalently coat paramagnetic beads to be used for IMS-qPCR. The IMS-qPCR method was able to specifically and sensitively detect C. jejuni in mixed cultures within 3 h.

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

    Directory of Open Access Journals (Sweden)

    Eleanor Watson

    2014-09-01

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

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

    DEFF Research Database (Denmark)

    Boysen, Louise; Wechter, Naja Strandby; Rosenquist, Hanne

    2013-01-01

    When assessing effects of decontamination techniques on counts of Campylobacter spp. on broiler meat, it is essential that the results reflect the variations that may exist. Decontamination studies often use high inoculation levels (107 to 108 cfu) and one or few strains of Campylobacter jejuni......, thereby restricting the results to reflect only a limited part of the true situation. This study presents results from physical and chemical decontamination of broiler meat medallions using different strains and different initial concentrations of C. jejuni. For 3 strains of C. jejuni, mean log reductions...... obtained by freezing at −20°C for 7 d was significantly higher for an initial concentration of 107 cfu/sample on the meat compared with an initial concentration of 103 cfu/sample. For freezing at −20°C for 24 h or application of 6% tartaric acid and subsequent storage for 24 h, no statistically significant...

  14. Influence of the Gut Microbiota Composition on Campylobacter jejuni Colonization in Chickens.

    Science.gov (United States)

    Han, Zifeng; Willer, Thomas; Li, Li; Pielsticker, Colin; Rychlik, Ivan; Velge, Philippe; Kaspers, Bernd; Rautenschlein, Silke

    2017-11-01

    The Campylobacter jejuni -host interaction may be affected by the host's gut microbiota through competitive exclusion, metabolites, or modification of the immune response. To understand this interaction, C. jejuni colonization and local immune responses were compared in chickens with different gut microbiota compositions. Birds were treated with an antibiotic cocktail (AT) (experiments 1 and 2) or raised under germfree (GF) conditions (experiment 3). At 18 days posthatch (dph), they were orally inoculated either with 10 4 CFU of C. jejuni or with diluent. Cecal as well as systemic C. jejuni colonization, T- and B-cell numbers in the gut, and gut-associated tissue were compared between the different groups. Significantly higher numbers of CFU of C. jejuni were detected in the cecal contents of AT and GF birds, with higher colonization rates in spleen, liver, and ileum, than in birds with a conventional gut microbiota ( P microbiota. Histopathological gut lesions were observed only in C. jejuni -inoculated AT and GF birds but not in microbiota-colonized C. jejuni -inoculated hatchmates. These results demonstrate that the gut microbiota may contribute to the control of C. jejuni colonization and prevent lesion development. Further studies are needed to identify key players of the gut microbiota and the mechanisms behind their protective role. Copyright © 2017 Han et al.

  15. Campylobacter

    NARCIS (Netherlands)

    Wagenaar, J.A.

    2015-01-01

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

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

    Science.gov (United States)

    Saiyudthong, S; Phusri, K; Buates, S

    2015-07-01

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

  17. Pengembangan Metode Imunokromatografi untuk Deteksi secara Serologi Campylobacter jejuni pada Ayam DEVELOPMENT OF IMMUNOCHROMATOGRAPHIC METHOD FOR DETECTING Campylobacter jejuni ON CHICKEN SEROLOGICALLY

    Directory of Open Access Journals (Sweden)

    Anwar Rosyidi

    2013-11-01

    Full Text Available Campylobacter spp cause infection in human in the form of gastroenteritis and Guillain-Barre syndrome.Campylobacter jejuni is one of  Campylobacter spp responsible for about 90% of cases of Campylobacteriosisin human with gastroenteritis.  Efforts to prevent the incidence of Campylobacteriosis in humans should bestarted with an assessment on its level of poultry. This study aims to develop a diagnostic test for C. jejunion chicken using immunochromatographic serological method. Stages of the research consist of thepreparation of test card, antigen preparation, optimization of the antigen and serum. The results showedthat immmunochromatographic card can distinguish infected serum from the uninfected.The minimumamount of antigen  was found to be 1,5 ng/µl. It needs 10 µl serum to perform the test properly.

  18. Beta-resorcylic acid, a phytophenolic compound, reduces Campylobacter jejuni in post-harvest poultry

    Science.gov (United States)

    Human Campylobacter infections, a leading foodborne illness globally, has been linked with the high prevalence of this bacterium on raw retail chicken products. Reduction of Campylobacter counts on poultry products would greatly reduce the risk of subsequent infections in humans. To this end, this s...

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

    DEFF Research Database (Denmark)

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

    2006-01-01

    safety. Bacteriological methods for determination of Campylobacter excretion level were combined with colony-blot hybridization and real-time PCR for specific detection of C. jejuni in pigs. Campylobacter was isolated from pigs (n = 47), paddock environment (n = 126) and wildlife (n = 44), identified...... to species by real-time PCR and sub-typed by serotyping (Penner) and pulse-field gel electrophorsis (PFGE) genotyping. All pigs excreted Campylobacter (10(3)-10(7) CFU g(-1) faeces) from the age of 8-13-weeks old. C jejuni was found in 29% of pigs in three consecutive trials and always in minority to C. coli......The occurrence and species distribution of thermophilic Campylobacter was investigated in organic outdoor pigs. An increased exposure of outdoor pigs to C jejuni from the environment may cause a shift from a normal dominance of C coli to more C jejuni, which may imply a concern of reduced food...

  20. Genetic diversity and antimicrobial resistance profiles of Campylobacter coli and Campylobacter jejuni isolated from broiler chicken in farms and at time of slaughter in central Italy.

    Science.gov (United States)

    Pergola, S; Franciosini, M P; Comitini, F; Ciani, M; De Luca, S; Bellucci, S; Menchetti, L; Casagrande Proietti, P

    2017-05-01

    Genetic diversity and antimicrobial resistance of Campylobacter coli and Campylobacter jejuni were investigated along the broiler chicken production chain in central Italy. Campylobacter sp. isolated from cloacal swabs in farms (n = 116) and from the neck skin of chilled and eviscerated carcasses at slaughter (n = 24) were identified as C. coli (n = 99) and C. jejuni (n = 41) by multiplex PCR. Characterization by single amplified fragment length polymorphism (s-AFLP) revealed a specific genotype of Campylobacter for each farm. Minimal inhibitory concentration showed high prevalence of fluoroquinolones (70%), tetracycline (70%) and erythromycin (30%) resistance among C. coli isolates. Campylobacter jejuni isolates showed lower prevalence of fluoroquinolone (39%) and tetracycline (10%) resistance, and all isolates were susceptible to erythromycin. The S-AFLP types of the C. coli and C. jejuni isolates were associated with their antimicrobial resistance profiles (P Campylobacter isolates suggested that a specific genotype was harboured in each farm. A considerable number of C. coli isolates were resistant to erythromycin. Campylobacter coli was detected more frequently than C. jejuni in contrast to common findings for poultry. The high prevalence of 30% resistance to erythromycin in C. coli strains isolated from poultry is worrisome, as this is the first antibiotic of choice to treat human campylobacteriosis. © 2017 The Society for Applied Microbiology.

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

    NARCIS (Netherlands)

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

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

    DEFF Research Database (Denmark)

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

    2010-01-01

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

  3. Prevalence of genetic determinants and phenotypic resistance to ciprofloxacin in Campylobacter jejuni from lithuania

    DEFF Research Database (Denmark)

    Aksomaitiene, Jurgita; Ramonaite, Sigita; Olsen, John E.

    2018-01-01

    Recently, the number of reports on isolation of ciprofloxacin resistant Campylobacter jejuni has increased worldwide. The aim of this study was to determine the prevalence of resistance to ciprofloxacin and its genetic determinants among C. jejuni isolated from humans (n = 100), poultry products (n...... = 96) and wild birds (n = 96) in Lithuania. 91.4% of the C. jejuni isolates were phenotypically resistant to ciprofloxacin. DNA sequence analyses of the gyrA gene from 292 isolates revealed that a change in amino acid sequence, Thr86Ile, was the main substition conferring resistance to ciprofloxacin...... forty-five C. jejuni isolates showed one or more silent mutations, and 32.4% of examined isolates possessed six silent mutations. In addition to the ciprofloxacin resistant isolates harboring only Thr86Ile point mutation (110 isolates), the current study identified resistant isolates (n = 101) harboring...

  4. Analysis of topoisomerase mutations in fluoroquinolone-resistant and -susceptible Campylobacter jejuni strains isolated in Senegal.

    Science.gov (United States)

    Kinana, Alfred Dieudonné; Cardinale, Eric; Bahsoun, Ibrahim; Tall, Fatou; Sire, Jean-Marie; Garin, Benoit; Boye, Cheikh Saad-Bouh; Dromigny, Jacques-Albert; Perrier-Gros-Claude, Jean-David

    2007-04-01

    In this study, topoisomerase mutations in ciprofloxacin-resistant and -susceptible Campylobacter jejuni were analysed by DNA sequencing. In certain ciprofloxacin-resistant C. jejuni, the mechanism of resistance was complex. The Thr86-Ala substitution in the GyrA protein appears to play a role in increasing the minimum inhibitory concentration of nalidixic acid only. In addition, isolates with this amino acid change and those resistant to quinolones but lacking a mutation in the GyrA quinolone resistance-determining region could be derived from two different clones. Based on gyrA and gyrB polymorphisms, C. jejuni isolates from the Dakar region of Senegal appeared to be less diverse than those from other countries. Moreover, C. jejuni isolates in Senegal appeared to differ from European isolates by lack of a silent mutation at codon 120 of the gyrA gene.

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

    Science.gov (United States)

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

    2015-01-01

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

  6. Methylation-dependent DNA discrimination in natural transformation ofCampylobacter jejuni.

    Science.gov (United States)

    Beauchamp, Jessica M; Leveque, Rhiannon M; Dawid, Suzanne; DiRita, Victor J

    2017-09-19

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

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

    Science.gov (United States)

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

    2011-06-01

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

  8. Complete Genome Sequences of Plasmid-Bearing Multidrug-Resistant Campylobacter jejuni and Campylobacter coli Strains with Type VI Secretion Systems, Isolated from Retail Turkey and Pork

    OpenAIRE

    Marasini, Daya; Fakhr, Mohamed K.

    2017-01-01

    ABSTRACT We report the complete genome sequences of multidrug-resistant Campylobacter jejuni and Campylobacter coli isolated from retail turkey and pork, respectively. The chromosomes of these two isolates contained type VI secretion system genes. The two isolates also harbored large plasmids with antimicrobial resistance genes possibly contributing to their multidrug resistance.

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

    Science.gov (United States)

    Zeng, Ximin; Ardeshna, Devarshi; Lin, Jun

    2015-07-01

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

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

    Directory of Open Access Journals (Sweden)

    Christopher J Day

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

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

    Science.gov (United States)

    Jackowska-Tracz, A; Tracz, M

    2015-01-01

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

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

    Science.gov (United States)

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

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

    DEFF Research Database (Denmark)

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

    2014-01-01

    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...

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

    Science.gov (United States)

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

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

    NARCIS (Netherlands)

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

    2006-01-01

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

  16. Identification of a protein glycosylation operon from Campylobacter jejuni JCM 2013 and its heterologous expression in Escherichia coli.

    Science.gov (United States)

    Srichaisupakit, Akkaraphol; Ohashi, Takao; Fujiyama, Kazuhito

    2014-09-01

    Campylobacter jejuni is a human enteropathogenic bacterium possessing an N-glycosylation system. In this work, a protein glycosylation (pgl) operon conferring prokaryotic N-glycosylation in C. jejuni JCM 2013 was cloned and identified. Fourteen open reading frames (ORFs) were found in the pgl operon. The operon organization was similar to that of C. jejuni NCTC 11168, with 98% and 99% identities in overall nucleotide sequence and amino acid sequence, respectively. The pgl operon was heterologously co-expressed with model protein CmeA in the Escherichia coli BL21 ΔwaaL mutant. The immuno- and lectin-blotting analysis indicated the protein glycosylation on the recombinant CmeA. In addition, to analyze the glycan composition, the recombinant CmeA was purified and subjected to in-gel trypsin digestion followed by mass spectrometry analysis. The mass spectrometry analysis showed the presence of the N-acetylhexosamine residue at the reducing end but not the predicted di-N-acetylbacillosamine (diNAcBac) residue. Further glycan structural study using the conventional fluorophore-labeling method revealed the GalNAcα-GalNAcα-(Hex-)HexNAc-HexNAc-HexNAc-HexNAc structure. Transcriptional analysis showed that UDP-diNAcBac synthases and diNAcBac transferase are transcribed but might not function in the constructed system. In conclusion, a pgl operon from C. jejuni JCM 2013 successfully functioned in E. coli, resulting in the observed prokaryotic glycosylation. Copyright © 2014 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

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

    Science.gov (United States)

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

    2013-01-01

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

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

    Directory of Open Access Journals (Sweden)

    Paola Ariganello

    2013-01-01

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

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

    Science.gov (United States)

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

    2013-02-22

    Six hundred and sixty one samples - primarily fresh chicken faeces - were processed to isolate wild type Campylobacter jejuni bacteriophages, via overlay agar methods using C. jejuni NCTC 12662. The aims of this study were to isolate and purify bacteriophages and then test for their ability to lyse field strains of C. jejuni in vitro. Of all samples processed, 130 were positive for bacteriophages. A distinct difference was observed between samples from different poultry enterprises. No bacteriophages could be isolated from indoor broilers. The majority of bacteriophages were isolated from free range poultry - both broilers and egg layers. Bacteriophages were purified and then selected for characterization based on their ability to produce clear lysis on plaque assay, as opposed to turbid plaques. Two hundred and forty one C. jejuni field isolates were tested for sensitivity to the bacteriophages. Lysis was graded subjectively and any minimal lysis was excluded. Using this system, 59.0% of the C. jejuni isolates showed significant sensitivity to at least one bacteriophage. The sensitivity to individual bacteriophages ranged from 10.0% to 32.5% of the C. jejuni isolates. Five bacteriophages were examined by electron microscopy and determined to belong to the Myoviridae family. The physical size, predicted genetic composition and genome size of the bacteriophages correlated well with other reported Campylobacter bacteriophages. The reasons for the observed difference between indoor broilers and free range poultry is unknown, but are postulated to be due to differences in the Campylobacter population in birds under different rearing conditions. Copyright © 2012 Elsevier B.V. All rights reserved.

  20. Update on Campylobacter jejuni vaccine development for preventing human campylobacteriosis.

    Science.gov (United States)

    Jagusztyn-Krynicka, Elzbieta Katarzyna; Łaniewski, Paweł; Wyszyńska, Agnieszka

    2009-05-01

    Campylobacteriosis constitutes a serious medical and socioeconomic problem worldwide. Rapidly increasing antibiotic resistance of bacterial strains compels us to develop alternative therapeutic strategies and to search for efficient immunoprophylactic methods. The vast majority of Campylobacter infections in developed countries occur as sporadic cases, mainly caused by eating undercooked Campylobacter-contaminated poultry. The most efficient strategy of decreasing the number of human Campylobacter infections is by implementing protective vaccinations for humans and/or chickens. Despite more than 10 years of research, an effective anti-Campylobacter vaccine has not been developed. This review highlights our increasing knowledge of Campylobacter interaction with host cells and focuses on recently published data describing the efficacy of anti-Campylobacter vaccine prototypes.

  1. Analysis of putative chemoreceptor proteins of Campylobacter jejuni

    DEFF Research Database (Denmark)

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

    Campylobacter jejuni is the primary food borne bacterial pathogen in the developed world. A very important reservoir for C. jejuni is the gut of chickens, which are colonized efficiently and commensally by this organism. Predominantly the mucus filled crypts of the lower gastrointestinal tract...... substances dispensed in filter discs on PBS softagar. Interestingly, strong attractions towards substances with low pH were observed repeatedly in the filter assay but not in the capillary assay. However, there were no striking chemotactic difference between the mutants and the parental strain, hence...

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

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    Alberto Fica

    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.

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

    Science.gov (United States)

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

  4. Distribution of serotypes of Campylobacter jejuni and C-coli from Danish patients, poultry, cattle and swine

    DEFF Research Database (Denmark)

    Nielsen, Eva Møller; Engberg, Jørgen; Madsen, Mogens

    1997-01-01

    The number of human cases of enteritis caused by Campylobacter jejuni and C. coli is increasing in Denmark and other European countries. No systematic typing has earlier been performed on Campylobacter isolates of Danish origin. The primary purpose of this study was to provide a serotype distribu......, and on this basis both could be major sources of human campylobacteriosis....

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

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    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

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

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    Maojun Zhang

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

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

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    Corander Jukka

    2010-07-01

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

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

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    Helen L Brown

    2015-07-01

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

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

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    Sanad Yasser M

    2011-12-01

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

  10. Prevalence and antimicrobial resistance of Campylobacter jejuni and Campylobacter coli isolated from children and environmental sources in urban and suburban areas.

    Science.gov (United States)

    Szczepanska, Bernadeta; Andrzejewska, Małgorzata; Spica, Dorota; Klawe, Jacek J

    2017-04-04

    Campylobacteriosis is a dominant bacterial cause of foodborne infection and is considered the main public health problem in Europe and many other countries worldwide. In the study lasting from 2011 to 2013 we compared the prevalence and antimicrobial resistance of Campylobacter jejuni and Campylobacter coli isolated from children, domestic animals, poultry meat and surface water in Northern Poland. During a 3-years study 1973 samples were analysed. The results proved the presence of Campylobacter spp. in 306 (15.5%) samples. The percentage of Campylobacter-positive samples differed among the sample types, from 0% (freshwater beaches) to 38.6% (poultry meat in 2011). Prevalence of Campylobacter spp. in children isolates was 9.6%. It decreased from 13.2% in 2011 to 8.0% in 2013. It should be highlighted with a particular concern that Campylobacter jejuni was detected in 20.0% of fountains. All children and poultry meat isolates were susceptible to azithromycin. Two C. coli (3.7%) and four C. jejuni (3.3%) isolated from poultry meat were resistant to erythromycin. The highest percentage of C. jejuni isolates with resistance to ciprofloxacin were found in samples from 80% dogs and 85% ponds. Among isolates resistant to two antimicrobials 74.7% C. jejuni and 59.2% C. coli isolates were resistant to ciprofloxacin as well as to tetracycline. Only one cat C. coli isolate was resistant to both azithromycin and erythromycin. One C. jejuni isolate from a fountain was resistant to four antimicrobial agents (erythromycin, azithromycin, tetracycline and ciprofloxacin). The study proved that surface water, poultry meat and pets constituted potential sources of Campylobacter to children. Fountains can be a direct source of children campylobacteriosis but can also pollute other environments with multidrug-resistant Campylobacter. The high resistance to some antimicrobials among the isolates may lead to increasing numbers of difficult-to-treat campylobacteriosis cases among children.

  11. Changes within the intestinal flora of broilers by colonisation with Campylobacter jejuni.

    Science.gov (United States)

    Sofka, Dmitri; Pfeifer, Agathe; Gleiss, Barbara; Paulsen, Peter; Hilbert, Friederike

    2015-01-01

    In most European countries human campylobacteriosis is the most important bacterial zoonotic foodborne infection. Chicken meat is considered the main source of infection. Since most strategies assessed so far, in reducing Campylobacter colonization in chickens or in the reduction of human disease, have not been very effective, new knowledge regarding Campylobacter's interaction with the host is needed. In this study we analysed fecal and cecal samples of five chicken flocks of different Austrian farms for the occurrence of Cjejuni and C. coli, and analysed the intestinal microbiota by PCR-SSCP, cultural detection of lactic acid bacteria, Enterococci, Staphylococci, Enterobacteriaceae, E. coli, and total aerobic colony counts. Furthermore ten chicken samples of cecal content of a flock during colonization with Campylobacter spp. was analysed by high throughput sequencing. With all three methods used we could detect a change within the microbiota caused by Cjejuni. Enumeration of different bacteria was significantly lower in fecal samples positive for C. jejuni, pointing out that a higher water content and thus, a preliminary stage of diarrhea might appear during Campylobacter colonization. By PCR-SSCP analysis the microbiota composition differed between colonized and non-colonized chicken fecal samples. This could also be detected in community analysis by high throughput sequencing, but this difference was only a tendency and not statistically significant. It can be concluded that C. jejuni is interacting with the intestinal microflora in their respective hosts and hence, this has to be taken into account when providing new strategies to combat Campylobacter colonization and disease.

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

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    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.

  13. Cleaning and disinfection programs against Campylobacter jejuni for broiler chickens: productive performance, microbiological assessment and characterization.

    Science.gov (United States)

    Castro Burbarelli, Maria Fernanda de; do Valle Polycarpo, Gustavo; Deliberali Lelis, Karoline; Granghelli, Carlos Alexandre; Carão de Pinho, Agatha Cristina; Ribeiro Almeida Queiroz, Sabrina; Fernandes, Andrezza Maria; Moro de Souza, Ricardo Luiz; Gaglianone Moro, Maria Estela; de Andrade Bordin, Roberto; de Albuquerque, Ricardo

    2017-09-01

    Detailed cleaning and disinfection programs aims to reduce infection pressure from microorganisms from one flock to the next. However, studies evaluating the benefits to poultry performance, the sanitary status of the facilities, and the sanitary quality of the meat are rarely found. Thus, this study was designed to evaluate 2 cleaning and disinfecting programs regarding their influence on productive performance, elimination of Campylobacter, and characterization of Campylobacter jejuni strains when applied to broiler chickens' facilities. Two subsequent flocks with 960 birds each were distributed into 32 pens containing 30 birds each. In the first, the whole flock was inoculated with a known strain of Campylobacter jejuni in order to contaminate the environment. In the second flock, performance and microbiological evaluations were done, characterizing an observational study between 2 cleaning and disinfection programs, regular and proposed. The regular program consisted of sweeping facilities, washing equipment and environment with water and neutral detergent. The proposed cleaning program consisted of dry and wet cleaning, application of 2 detergents (one acid and one basic) and 2 disinfectants (250 g/L glutaraldehyde and 185 g/L formaldehyde at 0.5% and 210 g/L para-chloro-meta-cresol at 4%). Total microorganism count in the environment and Campylobacter spp. identification were done for the microbiological assessment of the environment and carcasses. The positive samples were submitted to molecular identification of Campylobacter spp. and posterior genetic sequencing of the species identified as Campylobacter jejuni. The birds housed in the facilities and submitted to the proposed treatment had better performance when compared to the ones in the regular treatment, most likely because there was a smaller total microorganism count on the floor, walls, feeders and drinkers. The proposed program also resulted in a reduction of Campylobacter spp. on floors

  14. Quantifying Transmission of Campylobacter jejuni in Commercial Broiler Flocks

    NARCIS (Netherlands)

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

    2009-01-01

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

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

    DEFF Research Database (Denmark)

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

    2001-01-01

    The incidence of human Campylobacter jejuni and C. coli infections has increased markedly in many parts of the world in the last decade as has the number of quinolone-resistant and, to a lesser extent, macrolide-resistant Campylobacter strains causing infections. We review macrolide and quinolone...... maintained, but fluoroquinolones may now be of limited use in the empiric treatment of Campylobacter infections in many regions....

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

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    Katarzyna A Radomska

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

  17. Effect of butyrate and Lactobacillus GG on a butyrate receptor and transporter during Campylobacter jejuni exposure.

    Science.gov (United States)

    Cresci, Gail A M; Mayor, Paul C; Thompson, Stuart A

    2017-03-01

    Campylobacter jejuni frequently infects humans causing many gastrointestinal symptoms, fever, fatigue and several long-term debilitating diseases. Current treatment for campylobacteriosis includes rehydration and in some cases, antibiotic therapy. Probiotics are used to treat several gastrointestinal diseases. Butyrate is a short-chain fatty acid known to promote intestinal health. Interaction of butyrate with its respective receptor (HCAR2) and transporter (SLC5A8), both expressed in the intestine, is associated with water and electrolyte absorption as well as providing defense against colon cancer and inflammation. Alterations in gut microbiota influence the presence of HCAR2 and SLC5A8 in the intestine. We hypothesized that adherence and/or invasion of C. jejuni and alterations in HCAR2 and SLC5A8 expression would be minimized with butyrate or Lactobacillus GG (LGG) pretreatment of Caco-2 cells. We found that both C. jejuni adhesion but not invasion was reduced with butyrate pretreatment. While LGG pretreatment did not prevent C. jejuni adhesion, it did result in reduced invasion which was associated with altered cell supernate pH. Both butyrate and LGG protected HCAR2 and SLC5A8 protein expression following C. jejuni infection. These results suggest that the first stages of C. jejuni infection of Caco-2 cells may be minimized by LGG and butyrate pretreatment. © FEMS 2017. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  18. Passive and active immunity of broiler chickens against Campylobacter jejuni and ways of disease transmission

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    Silvia Ondrašovičová

    2012-01-01

    Full Text Available The study deals with passive and active immunity of fifty-three broiler chickens after infection with culture of Campylobacter jejuni. Potential transfer of infection by faecal-oral and aerogenic routes was also investigated. Cloacal swabs and ceacal content were analyzed microbiologically. Identification of C. jejuni was carried out by polymerase chain reaction. Observation of passive immunity of broilers from 3 days of age showed that no transfer of C. jejuni infection occurred up to 12 day post-infection (p.i.. Observations of active immunity in fourteen 21 days old chickens infected with C. jejuni showed that 6 chickens were positive on day 3 p.i. and all infected chickens were positive on day 5 p.i. Investigations of the transfer of C. jejuni by faecal-oral route revealed positivity in two broilers on day 3 p.i. and in all tested chickens on day 5 p.i. Aerogenic transfer of infection was not recorded. This was one of the first studies in our country dealing with passive and active immunity of broiler chickens against C. jejuni and spreading of this zoonotic disease.

  19. Cow's milk with active immunoglobulins against Campylobacter jejuni: effects of temperature on immunoglobulin activity.

    Science.gov (United States)

    Riera, Francisco; Alvarez, Alejandro; Espi, Alberto; Prieto, Miguel; de la Roza, Begoña; Vicente, Fernando

    2014-04-01

    Adult Holstein cows were injected with an antiserum against Campylobacter jejuni and immunoglobulin activities in vitro were determined in blood and milk several weeks after injection. The immunoactivity of immunoglobulins in milk was measured by an ELISA after different temperature-time treatments (60-91°C and 4-3600 s) at laboratory and pilot-plant scales. Kinetic and thermodynamic parameters were determined. An increase in immunoglobulin activity in milk was detected several days after injection. Optical densities increased by three- to seven-fold in this period. The activity started to decay 4-5 weeks after injection. Immunoglobulins maintained most of their in vitro activity under pasteurisation conditions (72°C and 15 s) and were denatured following first-order kinetics. The injection protocol applied allows milk with specific immunoglobulins against Campylobacter jejuni to be obtained. Traditional pasteurisation did not reduce this activity. © 2013 Society of Chemical Industry.

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

    Science.gov (United States)

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

    2016-10-01

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

  1. High genetic similarity of ciprofloxacin-resistant Campylobacter jejuni in central Europe

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    Jasna eKovač

    2015-10-01

    Full Text Available Campylobacteriosis is the leading zoonosis in the European Union with the majority of cases attributed to Campylobacter jejuni. Although the disease is usually self-limiting, some severe cases need to be treated with antibiotics, primarily macrolides and quinolones. However, the resistance to the latter is reaching alarming levels in most of the EU countries. To shed light on the expansion of antibiotic resistance in central Europe, we have investigated genetic similarity across 178 ciprofloxacin-resistant Campylobacter jejuni mostly isolates in Slovenia, Austria and Germany. We performed comparative genetic similarity analyses using allelic types of seven MLST housekeeping genes, and the single nucleotide polymorphisms of quinolone resistance determining region located within the DNA gyrase subunit A gene. This analysis revealed high genetic similarity of isolates from clonal complex ST-21 that carry gyrA allelic type 1 in all three of these central-European countries, suggesting these ciprofloxacin resistant isolates arise from a recent common ancestor.

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

    Science.gov (United States)

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

    2014-07-02

    We compared Bolton enrichment broth supplemented with antimicrobial triclosan (T-Bolton broth) and normal Bolton broth for the isolation of Campylobacter jejuni (C. jejuni) and Campylobacter coli (C. coli) from chicken carcass rinse. Whole chickens were rinsed with buffered peptone water prior to enrichment in normal Bolton broth or T-Bolton broth, followed by inoculation onto modified charcoal-cefoperazone-deoxycholate agar (mCCDA). Suspect colonies were confirmed by PCR. We observed a significantly higher number of C. jejuni or C. coli-positive samples in the T-Bolton broth (71.3%) than in the normal Bolton broth (27.5%) (p<0.05). Furthermore, the number of contaminated mCCDA plates was lower after enrichment in T-Bolton broth (3.8%) than in the normal Bolton broth (75%) (p<0.05), indicating that T-Bolton broth has higher selectivity. Finally, we identified extended-spectrum β-lactamase-producing Escherichia coli as the predominant competing flora in normal Bolton broth. In conclusion, the use of T-Bolton broth results in significant elimination of competing bacteria. Copyright © 2014 Elsevier B.V. All rights reserved.

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

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    Jenny Olofsson

    2015-09-01

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

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

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    Denis Martine

    2011-05-01

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

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

    Science.gov (United States)

    Leblanc-Maridor, Mily; Beaudeau, François; Seegers, Henri; Denis, Martine; Belloc, Catherine

    2011-05-22

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

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

    Science.gov (United States)

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

    2016-03-01

    Among zoonotic diseases, campylobacteriosis stands out as the major bacterial infection producing human gastroenteritis. Antimicrobial therapy, only recommended in critical cases, is challenged by resistance mechanisms that should be unambiguously detected for achievement of effective treatments. Quinolone (ciprofloxacin) resistance of Campylobacter jejuni and Campylobacter coli, the 2 main Campylobacter detected in humans, is conferred by the mutation gyrA C-257-T, which can be genotyped by several methods that require a previous identification of the pathogen species to circumvent the sequence polymorphism of the gene. A multiplex PCR, based on degenerated oligonucleotides, has been designed for unambiguous identification of the quinolone resistance determinant in Campylobacter spp. isolates. The method was verified with 249 Campylobacter strains isolated from humans (141 isolates) and from the 3 most important animal sources for this zoonosis: poultry (34 isolates), swine (38 isolates), and cattle (36 isolates). High resistance to ciprofloxacin, MIC above 4μg/mL, linked to the mutated genotype predicted by MAMA-DEG PCR (mismatch amplification mutation assay PCR with degenerated primers) was found frequently among isolates from the different hosts. Copyright © 2016 Elsevier Inc. All rights reserved.

  7. Studying the Phenotypic and Genotypic Expression of Antibiotic Resistance in Campylobacter jejuni under Stress Conditions.

    Science.gov (United States)

    Efimochkina, N R; Stetsenko, V V; Bykova, I V; Markova, Yu M; Polyanina, A S; Aleshkina, A I; Sheveleva, S A

    2018-03-01

    Specific features for the development of resistance in Campylobacter jejuni strains were studied after treatment with antibiotics of 6 pharmacological groups. Populations of 18 native strains of C. jejuni (isolated from raw poultry products) and their subcultures (obtained after 2-3-fold stress exposures to antimicrobial agents in subinhibitory doses) were examined to evaluate the expression of phenotypic antibiotic resistance. Genotypic properties of strains were studied by the PCR with primers that detect the presence of genes for resistance to aminoglycosides (aphA-1, aphA-3, and aphA-7), tetracyclines (tetO), and quinolones (GZgyrA). The majority of test strains of C. jejuni exhibited a high resistance to nalidixic acid, ciprofloxacin, and tetracycline, which reached the maximum value after numerous passages. The expression of antibiotic resistance was greatest in the presence of nalidixic acid and tetracycline. Ciprofloxacin resistance of 33% strains, which were initially resistant to this antibiotic, was increased after 2-3-fold treatment. We revealed a high degree of correspondence between phenotypic and genotypic profiles of antibiotic resistance in food isolates of Campylobacter. One, two, or more genes of aphA were identified in 85% strains phenotypically resistant to aminoglycosides. The tetO gene was found nearly in all strains resistant to tetracycline. Studying the biofilm matrix in C. jejuni after culturing with antibiotics in subinhibitory doses showed that quinolones (particularly nalidixic acid) and tetracyclines potentiate the formation of biofilms and increase the tolerance of Campylobacter to stress exposures. The intensity of biofilm growth was shown to depend little on the effect of macrolides and aminoglycosides. Therefore, the presence of these agents in residual concentrations is associated with a lower risk for the development of antibiotic resistance in C. jejuni populations.

  8. Functional characterization of a lipoprotein-encoding operon in Campylobacter jejuni.

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    Mayumi Oakland

    Full Text Available BACKGROUND: Bacterial lipoproteins have important functions in bacterial pathogenesis and physiology. In Campylobacter jejuni, a major foodborne pathogen causing gastroenteritis in humans, the majority of lipoproteins have not been functionally characterized. Previously, we showed by DNA microarray that CmeR, a transcriptional regulator repressing the expression of the multidrug efflux pump CmeABC, modulates the expression of a three-gene operon (cj0089, cj0090, and cj0091 encoding a cluster of lipoproteins in C. jejuni. METHODOLOGY/PRINCIPAL FINDINGS: In this work, we characterized the function and regulation of the cj0089-cj0090-cj0091 operon. In contrast to the repression of cmeABC, CmeR activates the expression of the lipoprotein genes and the regulation is confirmed by immunoblotting using anti-Cj0089 and anti-Cj0091 antibodies. Gel mobility shift assay showed that CmeR directly binds to the promoter of the lipoprotein operon, but the binding is much weaker compared with the promoter of cmeABC. Analysis of different cellular fractions indicated that Cj0089 was associated with the inner membrane, while Cj0091 was located on the outer membrane. Inactivation of cj0091, but not cj0089, significantly reduced the adherence of C. jejuni to INT 407 cells in vitro, indicating that Cj0091 has a function in adherence. When inoculated into chickens, the Cj0091 mutant also showed a defect in early colonization of the intestinal tract, suggesting that Cj0091 contributes to Campylobacter colonization in vivo. It was also shown that Cj0091 was produced and immunogenic in chickens that were naturally infected by C. jejuni. CONCLUSION/SIGNIFICANCE: These results indicate that the lipoprotein operon is subject to direct regulation by CmeR and that Cj0091 functions as an adhesion mechanism in C. jejuni and contributes to Campylobacter colonization of the intestinal tract in animal hosts.

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

    Science.gov (United States)

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

    2016-10-01

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

  10. Prevalence of Campylobacter jejuni, Campylobacter coli and enteric Helicobacter in domestic and free living birds in North-Western Italy.

    Science.gov (United States)

    Robino, P; Tomassone, L; Tramuta, C; Rodo, M; Giammarino, M; Vaschetti, G; Nebbia, P

    2010-09-01

    In order to investigate the prevalence of some thermophilic Campylobacter (C. jejuni and C. coli) and enteric Helicobacter (H. pullorum and H. canadensis) in domestic and wild birds, a total of 278 bird caecal samples were analyzed over a 2 year period in North-Western Italy. Samples were collected from poultry raised in intensive farming at the slaughterhouse (n=102, group A) and in small scale rural farms (n=60, group B) as well as from wild birds (n=116, group C). PCR amplifications were carried out on DNA extracted from caecal samples. Molecular assays targeted the hipO gene for C. jejuni, the asp gene for C. coli and the 16S rRNA gene of H. pullorum/H. canadensis. To differentiate H. pullorum from H. canadensis, PCR products were subjected to an ApaLI digestion assay. Prevalence of thermophilic Campylobacter and enteric Helicobacter was significantly different among groups (p<0.0001). Campylobacter infections were detected in all three bird groups (78.4% group A, 18.3% group B and 38.8% group C, respectively), Helicobacter infections were only detected in poultry, with H. pullorum infecting 68.6% of group A and 21.7% of group B birds. H. canadensis was detected in Guinea fowls (group A) and for the first time in pheasants (group B). Mixed infections by enteric Campylobacter and Helicobacter were shown in 53.9% of group A and in 5.0 % of group B. Our results show that both microorganisms commonly infect poultry, especially intensive farming animals. Only hooded crows among the wild bird group (group C), proved to be highly sensitive to Campylobacter infection.

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

    DEFF Research Database (Denmark)

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

    2006-01-01

    The effect of a Campylobacter jejuni colonization on the development of the microflora of the cecum and the ileum of broiler chickens was studied using molecular methods. The infection did affect the development and complexity of the microbial Communities of the ceca, but we found no permanent ef....... Some of these DGGE bands could be affiliated with Lactobacillus reuteri, Clostridium perfringens, and the genus Klebsiella....

  12. Characterization of a Plasmid-Encoded Type IV Secretion System in Campylobacter jejuni 81-176

    Science.gov (United States)

    2004-01-01

    Rutherford, A.H. van Vliet, S. Whitehead, and B.G. Barrell. 2000. The complete genome sequence of the food - borne pathogen Campylobacter jejuni reveals...G. A. Bradley, and L. A. Joens. 1993. Newborn piglet model for campylobacteriosis . Infect Immun 61:3466-3475. Backert, S., Ziska, E., Brinkmann, V...outbreak of campylobacteriosis associated with consumption of raw milk. J Infect Dis 152:592-596. Krause, S., M. Barcena, W. Pansegrau, R. Lurz, J. M

  13. Characterization of probiotic strains: an application as feed additives in poultry against Campylobacter jejuni.

    Science.gov (United States)

    Santini, Cecilia; Baffoni, Loredana; Gaggia, Francesca; Granata, Marta; Gasbarri, Rossana; Di Gioia, Diana; Biavati, Bruno

    2010-07-31

    Campylobacteriosis is at present the most frequent zoonosis in humans and the main source is poultry meat contaminated by Campylobacter jejuni. An alternative and effective approach to antibiotic administration to livestock to reduce bacterial contamination is the use of probiotics, which can help to improve the natural defence of animals against pathogenic bacteria. In this study 55 lactic acid bacteria and bifidobacteria were screened for desirable properties for their application as probiotics against Campylobacter in poultry. All bacteria were examined for their antimicrobial activity against three C. jejuni strains. Strains exhibiting the highest anti-Campylobacter activity were examined for their survival in the gastro intestinal tract (low pH and presence of bile salts) and food/feed processing conditions (high temperature, high NaCl concentration and starvation) and basic safety aspects such as antibiotic susceptibility and hemolytic activity were studied. On the basis of these activities, two strains, namely Lactobacillus plantarum PCS 20 and Bifidobacterium longum PCB 133, were chosen for an in vivo trial in poultry. They were separately administered to healthy chickens in order to evaluate their capability of colonizing the GI tract of poultry and to estimate their effect on C. jejuni population. The results evidenced that L. plantarum PCS 20 was not present in poultry feces at detectable concentration, whereas B. longum PCB 133 significantly increased after two weeks of daily administration and its amount was still high after a wash-out period of 6 days. In the same period, C. jejuni concentration in poultry feces was significantly reduced in chickens administered with B. longum PCB 133. Therefore, B. longum PCB 133, possessing interesting probiotic properties and a marked anti-Campylobacter activity both in vitro and in vivo, is an excellent candidate for being employed as additives to feed for poultry for the reduction of food-borne campylobacteriosis

  14. Phenotypic and Genotypic Detection of Campylobacter Jejuni at Local Chicken and Chicken Meat

    OpenAIRE

    Rosyidi, A; Budhiharta, S; Asmara, W; Yudhabuntara, D

    2010-01-01

    The Objective of this study was to identify the existence of Campylobacter jejuni based on phenotypic and genotypic characteristic in local chicken and chicken meats. Samples of local chicken intestine and meat were tested for the bacterial existence. Phenotypic examination was carried out by means of cultivation followed by gram staining and biochemical tests. Genotypic examination was conducted by polymerase chain reaction (PCR) using genus specific16S rRNA gene at 816 bp and membrane-...

  15. Quantitative Immunocapture PCR Assay for Detection of Campylobacter jejuni in Foods

    OpenAIRE

    Waller, David F.; Ogata, Steven A.

    2000-01-01

    The rapid detection of food-borne bacterial pathogens as part of a quality control program is necessary for the maintenance of a safe food supply. In this report, we present our findings for an immunocapture PCR method for the detection of Campylobacter jejuni in foods. The method permits direct detection of the pathogen without an enrichment step and can be performed in approximately 8 h. Assay results are quantitative, and one cell in a milliliter sample can be detected. Application of the ...

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

    Energy Technology Data Exchange (ETDEWEB)

    Kiehlbauch, J.A.

    1986-01-01

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

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

    DEFF Research Database (Denmark)

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

    2010-01-01

    The aim of this study was to determine whether marination of chicken meat in different food ingredients call be used to reduce populations of Campylobacter jejuni strains, were exposed to different organic acids (tartaric, acetic. lactic, malic, and citric acids) and food marinating ingredients...... 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...... of approximately 1.2 log units after 3 days of storage. Taste evaluations of chicken meat that had been marinated and then fried were graded positively for flavor and texture. Thus, Success was achieved ill creating I marinade with an acceptable taste that reduced the counts of C. jejuni....

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

    Science.gov (United States)

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

    2014-11-25

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

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

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    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.

  20. Roles of RpoN in the resistance of Campylobacter jejuni under various stress conditions

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    Ryu Sangryeol

    2011-09-01

    Full Text Available Abstract Background Campylobacter jejuni is a leading foodborne pathogen worldwide. Despite the fastidious nature of C. jejuni growth, increasing numbers of human campylobacteriosis suggest that C. jejuni may possess unique mechanisms to survive under various stress conditions. C. jejuni possesses only three sigma factors (FliA, RpoD, and RpoN and lacks stress-defense sigma factors. Since FliA and RpoD are dedicated to flagella synthesis and housekeeping, respectively, in this study, we investigated the role of RpoN in C. jejuni's defense against various stresses. Results Survivability of an rpoN mutant was compared with the wild-type C. jejuni under various stress conditions. While the growth of the rpoN mutant was as comparably as that of the wild type in shaking cultures, the rpoN mutant exhibited significant survival defects when cultured statically. The rpoN mutant was more sensitive to osmotic stress (0.8% NaCl with abnormally-elongated cell morphology. Compared to the wile type, the rpoN mutant was more susceptible to acid stress (pH 5 and more resistant to hydrogen peroxide. However, the rpoN mutation had little effect on the resistance of C. jejuni to alkaline pH, heat, cold and antimicrobials. Conclusions The results demonstrate that RpoN plays an important role in C. jejuni's defense against various stresses which this bacterial pathogen may encounter during transmission to and infection of humans.

  1. Survey of Campylobacter jejuni in retail chicken meat products by application of a quantitative PCR protocol.

    Science.gov (United States)

    Rantsiou, Kalliopi; Lamberti, Cristina; Cocolin, Luca

    2010-07-31

    Campylobacter-contaminated food products are currently the cause of the highest number of gastroenteritis cases in developed countries. Apart for biosafety measures at the primary production level, no other official control measures are currently in place for its control. This is partly due to the lack of quantitative data regarding the prevalence and contamination level of different food products by Campylobacter spp. that does not allow for quantitative risk assessment. PCR-based methods, applied without prior enrichment, in food samples circumvent limitations associated with the quantification of foodborne pathogens by traditional, culture-dependent methods. In this study, we report the development of a protocol, based on the amplification of the rpoB gene of Campylobacter jejuni, by quantitative PCR (qPCR), directly in food samples. The quantification limit of the protocol was determined to be in the order of 10 colony forming units (cfu)/g or ml of food sample. The optimized protocol was applied for the survey of C. jejuni in naturally contaminated poultry samples. In parallel, traditional sampling was also performed. A high percentage of samples (87%) resulted to be positive by qPCR, while no C. jejuni was detected by traditional analysis. Furthermore, important differences were observed in the detection by qPCR between samples before and after enrichment. Copyright 2010 Elsevier B.V. All rights reserved.

  2. Comparative antimicrobial activity of gatifloxacin tested against Campylobacter jejuni including fluoroquinolone-resistant clinical isolates.

    Science.gov (United States)

    Hayward, C L; Erwin, M E; Barrett, M S; Jones, R N

    1999-06-01

    Campylobacter jejuni is an important pathogen that causes gastroenteritis, as well as other disease states such as meningitis and septic arthritis. In this study, the Etest (AB BIODISK, Solna, Sweden) results were compared to a reference agar dilution method using gatifloxacin, a new 8-methoxyfluoroquinolone. A total of 53 strains of C. jejuni initially isolated from patients in California and Mexico were tested. Results demonstrated a high correlation (r = 0.88) between the two utilized in vitro dilution methods. In addition, gatifloxacin activity was compared to that of ciprofloxacin, metronidazole, amoxicillin, erythromycin, chloramphenicol, gentamicin, tetracycline, and trimethoprim/sulfamethoxazole using the Etest. Gatifloxacin (MIC90, 4 micrograms/ml) was approximately eight- to 16-fold more potent than ciprofloxacin (Mic90, > 32 micrograms/ml), a commonly used fluoroquinolone for Campylobacter infections. Eight strains highly resistant to ciprofloxacin (MIC90, > 32 micrograms/ml) were tested for cross resistance against the newer fluoroquinolones (gatifloxacin, levofloxacin, trovafloxacin) and the rank order of potency was: gatifloxacin (MIC50, 16 micrograms/ml) > trovafloxacin = levofloxacin (MIC50, > 32 micrograms/mL). However, only 25% ciprofloxacin-resistant strains were inhibited by < or = 1 microgram/mL of gatifloxacin or trovafloxacin. These results for gatifloxacin against C. jejuni strains must be further assessed in the context of in vivo trials before the clinical role of this new fluoroquinolone can be determined. The Etest appears to be a simple and precise susceptibility test method for testing C. jejuni isolates against fluoroquinolones and other alternative therapeutic agents.

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

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    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.

  4. Studying the prevalence of Campylobacter jejuni in adults with gastroenteritis from northwest of Iran

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    Ahmadreza Mobaien

    2016-12-01

    Full Text Available Objective: To investigate the prevalence of Campylobacter jejuni (C. jejuni in the patients with gastroenteritis. Methods: This descriptive and analytical study included all adult patients with acute diarrhea admitted to the University Hospital of Zanjan Province who were enrolled in a one-year period from 2013 to 2014. Stool samples were checked for white blood cells (WBC and lactoferrin, then samples with WBC ≤ 5 positive for lactoferrin were selected for amplification of mapA gene of C. jejuni by RT-PCR assay. Results: In this study, 864 patients (410 men and 454 women with acute diarrhea were enrolled, of which about 718 patients had WBC less than 5 and 146 patients had WBC more than 5 in the stool exam. All inflammatory diarrhea samples were tested for lactoferrin and 111 cases of the samples tested were positive for lactferrin. A total of 40 samples out of 111 were positive for C. jejuni by RT. Conclusions: The finding of this study showed that the prevalence of inflammatory diarrhea and diarrhea caused by Campylobacter in this study was high. This need for education and awareness in this area, as well as appropriate treatment is too important.

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

    Directory of Open Access Journals (Sweden)

    Harriet Whiley

    2016-12-01

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

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

    Science.gov (United States)

    Whiley, Harriet; McLean, Ryan; Ross, Kirstin

    2016-12-23

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

  7. Confirmed identification and toxin profiling of Campylobacter jejuni using a thermostabilized multiplex PCR formulation.

    Science.gov (United States)

    Ramachandran, Nitya; Ramlal, Shylaja; Batra, Harsh Vardhan

    2017-07-01

    Cytolethal distending toxin (CDT) producing Campylobacter jejuni species are one of the leading causes of human gastroenteritis worldwide. The main intent of the study was to develop a multiplex PCR assay for the confirmed identification and toxin profiling of C. jejuni. The genes targeted were rpo B as genus specific, hip O for species; cdt A, cdt B, cdt C encoding respective subunit proteins of CDT with Internal Amplification Control (IAC). To enhance its application as a pre-mixed ready-to-use format, the master mix of developed mPCR was dried by lyophilization and stability was assessed. Thermostabilized reagents showed stability of 1.5 months at room-temperature and upto six months at 4 °C without any loss of functionality. The assay was evaluated on a number of presumptive Campylobacter isolates along with biochemical tests. Results obtained indicated the accurate identification of C. jejuni by developed mPCR format in contrast to misconception associated with biochemical assays. The assay was also tested on spiked samples for its real-time utility. Altogether, the room-temperature storable and ready-to- use mPCR format developed in this study could be preferred for rapid detection and confirmed identification of toxigenic strains of C. jejuni in place of conventional biochemical assays. © 2017 APMIS. Published by John Wiley & Sons Ltd.

  8. Structural and functional properties of a truncated hemoglobin from a food-borne pathogen Campylobacter jejuni.

    Science.gov (United States)

    Lu, Changyuan; Egawa, Tsuyoshi; Wainwright, Laura M; Poole, Robert K; Yeh, Syun-Ru

    2007-05-04

    Campylobacter jejuni contains two hemoglobins, Cgb and Ctb. Cgb has been suggested to perform an NO detoxification reaction to protect the bacterium against NO attack. On the other hand, the physiological function of Ctb, a class III truncated hemoglobin, remains unclear. By using CO as a structural probe, resonance Raman data show that the distal heme pocket of Ctb exhibits a positive electrostatic potential. In addition, two ligand-related vibrational modes, nu(Fe-O(2)) and nu(O-O), were identified in the oxy derivative, with frequencies at 542 and 1132 cm(-1), respectively, suggesting the presence of an intertwined H-bonding network surrounding the heme-bound ligand, which accounts for its unusually high oxygen affinity (222 microm(-1)). Mutagenesis studies of various distal mutants suggest that the heme-bound dioxygen is stabilized by H-bonds donated from the Tyr(B10) and Trp(G8) residues, which are highly conserved in the class III truncated hemoglobins; furthermore, an additional H-bond donated from the His(E7) to the Tyr(B10) further regulates these H-bonding interactions by restricting the conformational freedom of the phenolic side chain of the Tyr(B10). Taken together, the data suggest that it is the intricate balance of the H-bonding interactions that determines the unique ligand binding properties of Ctb. The extremely high oxygen affinity of Ctb makes it unlikely to function as an oxygen transporter; on the other hand, the distal heme environment of Ctb is surprisingly similar to that of cytochrome c peroxidase, suggesting a role of Ctb in performing a peroxidase or P450-type of oxygen chemistry.

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

    Directory of Open Access Journals (Sweden)

    Lieke B van Alphen

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

  10. Development and application of a real-time polymerase chain reaction method for Campylobacter jejuni detection.

    Science.gov (United States)

    Zhang, Mao-Jun; Qiao, Bo; Xu, Xue-Bin; Zhang, Jian-Zhong

    2013-05-28

    To develop a real-time polymerase chain reaction (PCR) method to detect and quantify Campylobacter jejuni (C. jejuni) from stool specimens. Primers and a probe for real-time PCR were designed based on the specific DNA sequence of the hipO gene in C. jejuni. The specificity of the primers and probe were tested against a set of Campylobacter spp. and other enteric pathogens. The optimal PCR conditions were determined by testing a series of conditions with standard a C. jejuni template. The detection limits were obtained using purified DNA from bacterial culture and extracted DNA from the stool specimen. Two hundred and forty-two specimens were analyzed for the presence of C. jejuni by direct bacterial culture and real-time PCR. The optimal PCR system was determined using reference DNA templates, 1 × uracil-DNA glycosylase, 3.5 mmol/L MgCl2, 1.25 U platinum Taq polymerase, 0.4 mmol/L PCR nucleotide mix, 0.48 μmol/L of each primer, 0.2 μmol/L of probe and 2 μL of DNA template in a final volume of 25 μL. The PCR reaction was carried as follows: 95 °C for 4 min, followed by 45 cycles of 10 s at 95 °C and 30 s at 59 °C. The detection limit was 4.3 CFU/mL using purified DNA from bacterial culture and 10(3) CFU/g using DNA from stool specimens. Twenty (8.3%, 20/242) C. jejuni strains were isolated from bacterial culture, while 41 (16.9%, 41/242) samples were found to be positive by real-time PCR. DNA sequencing of the PCR product indicated the presence of C. jejuni in the specimen. One mixed infection of C. jejuni and Salmonella was detected in one specimen and the PCR test for this specimen was positive. The sensitivity of detection of C. jejuni from stool specimens was much higher using this PCR assay than using the direct culture method.

  11. Epidemiological and serological investigation of a waterborne Campylobacter jejuni outbreak in a Danish town.

    Science.gov (United States)

    Kuhn, K Gaardbo; Falkenhorst, G; Emborg, H-D; Ceper, T; Torpdahl, M; Krogfelt, K A; Ethelberg, S; Mølbak, K

    2017-03-01

    Following an unusually heavy rainfall in June 2009, a community-wide outbreak of Campylobacter gastroenteritis occurred in a small Danish town. The outbreak investigation consisted of (1) a cohort study using an e-questionnaire of disease determinants, (2) microbiological study of stool samples, (3) serological study of blood samples from cases and asymptomatic members of case households, and (4) environmental analyses of the water distribution system. The questionnaire study identified 163 cases (respondent attack rate 16%). Results showed a significant dose-response relationship between consumption of tap water and risk of gastroenteritis. Campylobacter jejuni belonging to two related flaA types were isolated from stool samples. Serum antibody levels against Campylobacter were significantly higher in cases than in asymptomatic persons. Water samples were positive for coliform bacteria, and the likely mode of contamination was found to be surface water leaking into the drinking-water system. This geographically constrained outbreak presented an ideal opportunity to study the serological response in persons involved in a Campylobacter outbreak. The serology indicated that asymptomatic persons from the same household may have been exposed, during the outbreak period, to Campylobacter at doses that did not elicit symptoms or alternatively had been exposed to Campylobacter at a time prior to the outbreak, resulting in residual immunity and thus absence of clinical signs.

  12. Campylobacter fetus ssp jejuni en patología humana

    Directory of Open Access Journals (Sweden)

    Luz Echeverri

    1982-06-01

    Full Text Available Algunos microorganismos, como los Rotovirus y el Compylobocter fetus ssp jejuni reconocidos y estudiados recientemente, han llegado a ocupar un lugar preponderante en el grupo de enteropatógenos considerados como problema en salud pública.

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

    DEFF Research Database (Denmark)

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

    2006-01-01

    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...... to be invasive in orally challenged chickens as well as in three different human epithelial cell lines.......-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...

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

    Directory of Open Access Journals (Sweden)

    Annika eFlint

    2010-11-01

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

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

    LENUS (Irish Health Repository)

    Corcionivoschi, N

    2012-02-01

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

  16. Comparison of proteomics profiles of Campylobacter jejuni strain Bf under microaerobic and aerobic conditions

    Directory of Open Access Journals (Sweden)

    Ramila Christiane Rodrigues

    2016-10-01

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

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

    Directory of Open Access Journals (Sweden)

    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.

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

    Science.gov (United States)

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

    2014-07-14

    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.

  19. Assessment of chicken protection against Campylobacter jejuni infection by immunization with avirulent Salmonella enterica sv. Typhimurium strain producing Campylobacter CjaD/Pal protein

    Directory of Open Access Journals (Sweden)

    Łaniewski P

    2012-08-01

    Full Text Available Pawel Laniewski,1 Malgorzata Lis,2 Agnieszka Wyszynska,1 Pawel Majewski,3 Renata Godlewska,1 Elzbieta Katarzyna Jagusztyn-Krynicka11Department of Bacterial Genetics, Institute of Microbiology, Faculty of Biology, University of Warsaw, Warsaw, Poland; 2Biowet Pulawy Ltd, Pulawy, Poland; 3Department of Vertebrate Physiology, Institute of Zoology, Faculty of Biology, University of Warsaw, Warsaw, PolandAbstract: Campylobacter jejuni is a major food-borne pathogen, causing gastroenteritis worldwide. Chickens are considered to be one of the most common sources of human C. jejuni infection in developed countries. Campylobacter CjaD/Pal protein (annotated as Cj0113 in C. jejuni strain NCTC11168 is a highly immunogenic, membrane-located antigen, conserved among different strains, with the potential to provide broad protection against C. jejuni colonization. The present study examines the immunogenicity and the general efficacy of avirulent S. enterica sv. Typhimurium Δcrp Δcya expressing C. jejuni CjaD as a chicken vaccine against Campylobacter colonization. The high copy number plasmid pYA3341 Asd+ was used as a cloning vector. Here, 1- and 14-day old chickens were orally immunized with a delivery vector strain, expressing C. jejuni CjaD. Two weeks later, they were challenged with a wild-type C. jejuni strain isolated from chicken carcasses. This schedule of immunization induced significant levels of serum-specific IgG as well as mucosal intestinal sIgA as measured by ELISA tests using Campylobacter membrane proteins as a coating antigen. Nevertheless, protection experiments did not result in significant reduction of colonization of vaccinated birds relative to nonvaccinated birds.Keywords: Campylobacter, cjaD, immunization, Pal

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

    Science.gov (United States)

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

    2017-06-01

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

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

    Directory of Open Access Journals (Sweden)

    Gisela González-Hein

    2013-12-01

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

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

    Directory of Open Access Journals (Sweden)

    Michael E Taveirne

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

  3. Mutational and transcriptomic changes involved in the development of macrolide resistance in Campylobacter jejuni.

    Science.gov (United States)

    Hao, Haihong; Yuan, Zonghui; Shen, Zhangqi; Han, Jing; Sahin, Orhan; Liu, Peng; Zhang, Qijing

    2013-03-01

    Macrolide antibiotics are important for clinical treatment of infections caused by Campylobacter jejuni. Development of resistance to this class of antibiotics in Campylobacter is a complex process, and the dynamic molecular changes involved in this process remain poorly defined. Multiple lineages of macrolide-resistant mutants were selected by stepwise exposure of C. jejuni to escalating doses of erythromycin or tylosin. Mutations in target genes were determined by DNA sequencing, and the dynamic changes in the expression of antibiotic efflux transporters and the transcriptome of C. jejuni were examined by real-time reverse transcription-PCR, immunoblotting, and DNA microarray analysis. Multiple types of mutations in ribosomal proteins L4 and L22 occurred early during stepwise selection. On the contrary, the mutations in the 23S rRNA gene, mediating high resistance to macrolides, were observed only in the late-stage mutants. Upregulation of antibiotic efflux genes was observed in the intermediately resistant mutants, and the magnitude of upregulation declined with the occurrence of mutations in the 23S rRNA gene. DNA microarray analysis revealed the differential expression of 265 genes, most of which occurred in the intermediate mutant, including the upregulation of genes encoding ribosomal proteins and the downregulation of genes involved in energy metabolism and motility. These results indicate (i) that mutations in L4 and L22 along with temporal overexpression of antibiotic efflux genes precede and may facilitate the development of high-level macrolide resistance and (ii) that the development of macrolide resistance affects the pathways important for physiology and metabolism in C. jejuni, providing an explanation for the reduced fitness of macrolide-resistant Campylobacter.

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

    Science.gov (United States)

    Sylte, M J; Inbody, M H; Johnson, T A; Looft, T; Line, J E

    2018-03-05

    Consumption of contaminated poultry products is the main source of human campylobacteriosis, for which Campylobacter jejuni is responsible for 90% of human cases. Although chickens are believed to be a main source of human exposure to C. jejuni, turkeys also contribute to cases of human infection. Little is known about the kinetics of C. jejuni intestinal colonization in turkeys, or best selective media for their recovery. Enumeration of C. jejuni from intestinal samples can be challenging because most selective Campylobacter media support the growth of non-Campylobacter organisms. In this study, we sought to compare a) C. jejuni isolates that persistently colonize different compartments of the poult intestinal tract, and b) selective media to enumerate C. jejuni from turkey intestinal samples. Three-week-old poults were orally colonized with C. jejuni isolates NCTC 11168 or NADC 20827 (isolated from a turkey flock). Mock-colonized poults were orally gavaged with uninoculated media. Poults were euthanized at d 3, 7, and 21 post colonization and direct plated on different selective Campylobacter media [Campy Line agar with sulfamethoxazole (CLA-S), CHROMagar Campylobacter (CAC) and Campy Cefex] for enumeration. Isolates NCTC 11168 and NADC 20827 poorly colonized the distal ileum. Both isolates colonized the colon, but the number of NADC 20827 significantly decreased at d 21. Isolates NCTC 11168 and NADC 20827 persistently colonized the cecum for up to 21 days. There was no significant difference in the Campylobacter amount recovered on CLA-S and CAC. Campy Cefex failed to prevent growth of background microbes to enumerate C. jejuni from turkey samples. Two independent PCR assays (multiplex PCR and qPCR) confirmed that colonies grown on CLA-S or CAC were C. jejuni. Data from this study demonstrated that isolates NCTC 11168 and NADC 20827 persistently colonized the cecum, and CLA-S or CAC were successful to enumerate Campylobacter from intestinal samples. These

  5. Quantitative detection of Campylobacter jejuni on fresh chicken carcasses by real-time PCR.

    Science.gov (United States)

    Rönner, Anna-Clara; Lindmark, Hans

    2007-06-01

    Campylobacter jejuni infection is a significant cause of foodborne gastroenteritis worldwide. Consumption and handling of poultry products is believed to be the primary risk factor for campylobacteriosis. Risk assessments require quantitative data, and C. jejuni is enumerated usually by direct plating, which sometimes allows growth of non-Campylobacter bacteria. The objective of the present study was to develop a quantitative real-time PCR method (q-PCR) for enumerating C. jejuni in chicken rinse without a culturing step. The procedure to obtain the template for the PCR assay involved (i) filtration of 10 ml of chicken rinse, (ii) centrifugation of the sample, and (iii) total DNA extraction from the pellet obtained using a commercial DNA extraction kit. The detection limit of the method was comparable to that for plating 100 microl of chicken rinse on modified charcoal cefoperazone deoxycholate agar, and the detection limit could be further improved 10-fold by concentrating the DNA eluate by ethanol precipitation. A close correlation for spiked chicken rinse was obtained for the results of the quantitative real-time PCR method and direct plating (r = 0.99). The coefficient of correlation for the methods was 0.87 when samples from chicken carcasses on the slaughter line were analyzed, whereas a lower correlation (r = 0.76) was obtained when samples from retail carcasses were analyzed. Greater variation in the proportion of dead and/or viable but not culturable Campylobacter types in the retail samples may explain the decreased correlation between the methods. Overall, the new method is simple and fast and the results obtained are closely correlated with those for direct plating for samples containing a low proportion of dead Campylobacter cells.

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

    Science.gov (United States)

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

    2007-09-28

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

  7. Antibiotic resistance modulation and modes of action of (--α-pinene in Campylobacter jejuni.

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    Jasna Kovač

    Full Text Available The aim of the study was to investigate the mode of action of (--α-pinene in terms of its modulation of antibiotic resistance in Campylobacter jejuni. Broth microdilution and ethidium bromide accumulation assays were used to evaluate the (--α-pinene antimicrobial activity, modulation of antimicrobial resistance, and inhibition of antimicrobial efflux. The target antimicrobial efflux systems were identified using an insertion mutagenesis approach, and C. jejuni adaptation to (--α-pinene was evaluated using DNA microarrays. Knock-out mutants of the key up-regulated transcriptional regulators hspR and hrcA were constructed to investigate their roles in C. jejuni adaptation to several stress factors, including osmolytes, and pH, using Biolog phenotypical microarrays. Our data demonstrate that (--α-pinene efficiently modulates antibiotic resistance in C. jejuni by decreasing the minimum inhibitory concentrations of ciprofloxacin, erythromycin and triclosan by up to 512-fold. Furthermore, (--α-pinene promotes increased expression of cmeABC and another putative antimicrobial efflux gene, Cj1687. The ethidium bromide accumulation was greater in the wild-type strain than in the antimicrobial efflux mutant strains, which indicates that these antimicrobial efflux systems are a target of action of (--α-pinene. Additionally, (--α-pinene decreases membrane integrity, which suggests that enhanced microbial influx is a secondary mode of action of (--α-pinene. Transcriptomic analysis indicated that (--α-pinene disrupts multiple metabolic pathways, and particularly those involved in heat-shock responses. Thus, (--α-pinene has significant activity in the modulation of antibiotic resistance in C. jejuni, which appears to be mediated by multiple mechanisms that include inhibition of microbial efflux, decreased membrane integrity, and metabolic disruption. These data warrant further studies on (--α-pinene to develop its use in the control of antibiotic

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

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    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.

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

    OpenAIRE

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

    1999-01-01

    A rapid and sensitive assay was developed for detection of small numbers of Campylobacter jejuni and Campylobacter coli cells in environmental water, sewage, and food samples. Water and sewage samples were filtered, and the filters were enriched overnight in a nonselective medium. The enrichment cultures were prepared for PCR by a rapid and simple procedure consisting of centrifugation, proteinase K treatment, and boiling. A seminested PCR based on specific amplification of the intergenic seq...

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

    Science.gov (United States)

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

    2016-01-01

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

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

    Science.gov (United States)

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

    2017-04-01

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

  12. Evaluation of transport and storage techniques for isolation of Campylobacter fetus subsp. jejuni from turkey cecal specimens.

    OpenAIRE

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

    1981-01-01

    Immediate culturing of fecal specimens is not always possible, and appropriate methods for transport and storage of Campylobacter fetus subsp. jejuni specimens have not been fully evaluated. Using nine techniques, we studied the survival of C. fetus subsp. jejuni in cecal specimens from infected turkeys. The organisms survived in specimens held without transport medium for 3 to 15 days (median, 9 days) at 4 degrees C, and 2 to 9 days (median, 4 days) at 25 degrees C. Only 20% of specimens fro...

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

    OpenAIRE

    Danielle Karen Narcizo; Maike Taís Maziero Montanhini

    2014-01-01

    Campylobacter jejuni is one of the most bacteria that cause food-borne diseases worldwide. Campylobacteriosis is commonly associated with the consumption of poultry meat; however, several outbreaks of the disease have also been linked to the ingestion of milk, usually due to failures in the pasteurization or even by raw milk’s consumption. This study aimed to evaluate the occurrence of C. jejuni in samples of raw or pasteurized milk, as well as evaluate different methodologies for analysis of...

  14. The Bacterial Species Campylobacter jejuni Induce Diverse Innate Immune Responses in Human and Avian Intestinal Epithelial Cells

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    Daniel A. John

    2017-09-01

    Full Text Available Campylobacter remain the major cause of human gastroenteritis in the Developed World causing a significant burden to health services. Campylobacter are pathogens in humans and chickens, although differences in mechanistic understanding are incomplete, in part because phenotypic strain diversity creates inconsistent findings. Here, we took Campylobacter jejuni isolates (n = 100 from multi-locus sequence typed collections to assess their pathogenic diversity, through their inflammatory, cytotoxicity, adhesion, invasion and signaling responses in a high-throughput model using avian and human intestinal epithelial cells. C. jejuni induced IL-8 and CXCLi1/2 in human and avian epithelial cells, respectively, in a MAP kinase-dependent manner. In contrast, IL-10 responses in both cell types were PI 3-kinase/Akt-dependent. C. jejuni strains showed diverse levels of invasion with high invasion dependent on MAP kinase signaling in both cell lines. C. jejuni induced diverse cytotoxic responses in both cell lines with cdt-positive isolates showing significantly higher toxicity. Blockade of endocytic pathways suggested that invasion by C. jejuni was clathrin- and dynamin-dependent but caveolae- independent in both cells. In contrast, IL-8 (and CXCLi1/2 production was dependent on clathrin, dynamin, and caveolae. This study is important because of its scale, and the data produced, suggesting that avian and human epithelial cells use similar innate immune pathways where the magnitude of the response is determined by the phenotypic diversity of the Campylobacter species.

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

    Science.gov (United States)

    Bui, Xuan Thanh; Wolff, Anders; Madsen, Mogens; Bang, Dang Duong

    2012-01-01

    Campylobacter spp. is the most common cause of bacterial diarrhoea in humans worldwide. Therefore, rapid and reliable methods for detection 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 quantification of viable Campylobacter jejuni directly from chicken faecal samples. The results of this method and a DNA-based quantitative real-time PCR (qPCR) method were compared with those of a bacterial culture method. Using bacterial culture and RT-qPCR methods, viable C. jejuni cells could be detected for up to 5 days in both the C. jejuni spiked and the naturally contaminated faecal samples. We found that no RT-qPCR signals were obtained when viable C. jejuni cells could not be counted by the culture method. In contrast, using a DNA-based qPCR method, dead or non-viable Campylobacter cells were detected, and all tested samples were positive, even after 20 days of storage. The developed method for detection and quantification of viable C. jejuni cells directly from chicken faecal samples can be used for further research on the survival of Campylobacter in the environment. Copyright © 2011 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.

  16. Correlation between flagellin A (flaA) genotypes and antimicrobial susceptibility patterns of Campylobacter jejuni strains isolated from children with gastroenteritis in Athens, Greece.

    Science.gov (United States)

    Ioannidis, Anastassios; Nicolaou, Chrysoula; Chatzipanagiotou, Stylianos

    2009-12-01

    Campylobacter jejuni is one of the most common enteric pathogens worldwide. The bacterium is transmitted to humans via contaminated food and water. In the majority of cases the disease is self-limiting, but treatment is indicated in immunocompromised patents, in severe cases with septicemia, and in children. The subtyping of clinical, animal, and food C. jejuni isolates is very important for epidemiological studies. In the present study, 192 Campylobacter jejuni isolates characterized by pulsed-field gel electrophoresis (PFGE) of SmaI digested genomic DNA were further examined with respect to their antimicrobial resistance and their flagellin A (flaA) genotypes in order to disclose any correlation between a certain flaA type and a specific antimicrobial susceptibility pattern. C. jejuni clinical isolates were collected from infected children up to 14 years of age from five general hospitals in the area of Attica, Greece, during the period 2004-7. C. jejuni strain isolation and identification from stool samples were performed by conventional bacteriological methods. SmaI restriction fragments were prepared as described previously for the PFGE analysis. Antimicrobial susceptibility was tested and interpreted by determination of the minimal inhibitory concentration (MIC) by use of the agar dilution method as described by the Clinical Laboratory Standards Institute. FlaA typing was performed by PCR amplification of the corresponding gene, and the product was digested with DdeI and visualized by agarose gel electrophoresis. Data were analyzed using the software Gene Profiler 1-D Gel Analysis and Data Basing for Windows((R)). A statistically significant correlation between certain flaA genotypes (flaA 17 Greece [GR], flaA 19 GR and flaA 39 GR) and resistance to some antimicrobial agents (ampicillin, amoxicillin/clavulinic acid [co-amoxiclav], erythromycin, nalidixic acid, and ciprofloxacin) was detected in C. jejuni strains isolated from infected children. Further

  17. Aeromonas hydrophila and Campylobacter jejuni isolated in fresh tuna (Thunnus spp.) sold in São Paulo, Brazil

    OpenAIRE

    Andréa Moura Costa; Aline Feola Carvalho; Rodrigo César Fredrigo; Patrícia Freitas Kobayashi; Eliana Scarcelli Pinheiro

    2016-01-01

    Aeromonas hydrophila e Campylobacter jejuni são bactérias de importância emergente em saúde pública, porém com escassos trabalhos publicados na área de pescado. O presente estudo investigou a presença de Aeromonas hydrophila e Campylobacter jejuni em amostras de atum (Thunnus spp.) fresco, capturados no litoral de Santa Catarina e distribuídos no comércio atacadista de São Paulo, SP. Foram colhidas 85 amostras de filé de atum e processadas por análises bacteriológicas e PCR. Do total, 11/85 (...

  18. Campylobacter jejuni infection and IgE sensitization in up to 2-year-old infants

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    Jovanović Dara

    2015-01-01

    Full Text Available Background/Aim. The “hygiene hypothesis” addresses the correlation between the occurrence of atopy and the frequency of infections in the earliest age, explaining an increase in the incidence of atopic diseases by living in good, infection-free, hygienic conditions. The aim of our study was to determine the conection between atopy and Campylobacter infection, and to analyze the association between serum concentrations of total IgE and Campylobacter infection in relation to atopy in children up to two years. Methods. A case control study was conducted with the sample of 98 infants of the average age of 8 months. Total serum IgE and Phadiatop infant multi-test were determined on Immunocap-100 (Phadia AB, Uppsala, Sweden. The presence of atopy was determined by detection of serumspecific IgE ≥ 0.35 kUA/L (Phadiatop infant positive and serum IgM, IgA, IgG levels against C. jejuni were determined by a quantitative immuno-enzyme test - SERION ELISA classic. Results. Total IgE cut-off values ≥ 15 kU/L point to atopy in infants, and tIgE cut-off values ≥ 8.1 kU/L pointed to a C. jejuni infection in infants. Within the group of atopic children, tIgE levels ≥ 29.8 kU/L point to C. jejuni infection, and within the group of non-atopic children, tIgE levels ≥ 5.9 kU/L point to infection. Enteritis is not a predictor of C. jejuni infection, because of a high frequency of asymptomatic cases of infection. The risk factors for C. jejuni infection are age and tIgE, and the protective factors are breastfeeding and atopy. Conclusion. C. jejuni infection increases the total serum IgE level, which is predictive of infection, regardless of the presence of atopy. The presence of symptomatic C. jejuni infection reduces the risk of atopy in a child of the age of 5-24 months by the factor of 10.

  19. Antibodies, directed towards Campylobacter jejuni antigens, in sera from poultry abattoir workers

    Science.gov (United States)

    Cawthraw, S A; Lind, L; Kaijser, B; Newell, D G

    2000-01-01

    Occupational exposure of susceptible humans to Campylobacter jejuni appears to result in resistance to disease. This is believed to be due to acquired protective immunity. To support this hypothesis the levels of C. jejuni-specific IgG and IgM antibodies were determined in sera from poultry abattoir workers. Such individuals are persistently exposed to C. jejuni, but apparently rarely acquire campylobacteriosis. Sera from 43 short-term workers (employed ≤1 month), 78 long-term workers and 40 blood donors were investigated by ELISA. In 51 individuals a second serum sample, taken at least 1 month after the first, was also investigated. Eight workers had C. jejuni-positive faecal cultures and only one, a short-term worker, had symptoms of campylobacteriosis. There were significantly higher levels of specific IgG antibodies in long-term workers than in either of the other groups. There was no significant difference detectable in specific IgM antibody levels between any of the groups. The results provide supporting evidence that long-term exposure to C. jejuni induces circulating antibodies which reflect apparent reduced susceptibility to disease. Western blotting showed flagellin and polypeptides of 45, 40, 32 and 30 kD bound antibodies significantly more frequently by sera from long-term workers than short-term workers and blood donors. The most commonly detected antigens were the 40-kD (80%) and flagellin (55%). The results indicate that specific serum IgG responses induced by endemic exposure to C. jejuni might be directed towards a small number of protein antigens with apparently conserved epitopes. PMID:11012618

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

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    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

  1. Detection of Campylobacter jejuni in naturally contaminated chicken skin by melting peak analysis of amplicons in real-time PCR.

    Science.gov (United States)

    Oliveira, Tereza C R M; Barbut, Shai; Griffiths, Mansel W

    2005-09-25

    Contamination of poultry by Campylobacter spp. is a significant source of human diarrheal diseases. Traditional methods currently used to detect Campylobacter in foods are time-consuming and labor-intensive. In this study, primers designed for the Campylobacter jejuni cadF gene sequence were used in a SYBR Green I real-time PCR assay as an alternative to a conventional bacteriological method for the rapid detection of C. jejuni from poultry. Twelve portions of chicken purchased from two local grocery stores and 39 portions obtained from a commercial processing plant were examined. Samples of the skin were enriched in Bolton broth at 37 degrees C for 3 h and then at 42 degrees C for 9, 21, or 45 h under microaerobic conditions. DNA was extracted from 1-ml aliquots of the enrichment cultures using 1% Triton X-100. The DNA was used as the template in a real-time polymerase chain reaction (PCR) assay. After 24 h of enrichment, C. jejuni was isolated from 13 samples and all of the positive cultures were also detected by the real-time PCR procedure. C. jejuni was detected by both methods from samples artificially contaminated with 1 or 10 CFU of C. jejuni per 10 g, after 24 h of enrichment. The real-time PCR method was found to be sensitive and specific. It significantly reduced the time required for the detection of C. jejuni in poultry following enrichment of samples.

  2. Aeromonas hydrophila and Campylobacter jejuni isolated in fresh tuna (Thunnus spp. sold in São Paulo, Brazil

    Directory of Open Access Journals (Sweden)

    Andréa Moura Costa

    2016-04-01

    Full Text Available Aeromonas hydrophila e Campylobacter jejuni são bactérias de importância emergente em saúde pública, porém com escassos trabalhos publicados na área de pescado. O presente estudo investigou a presença de Aeromonas hydrophila e Campylobacter jejuni em amostras de atum (Thunnus spp. fresco, capturados no litoral de Santa Catarina e distribuídos no comércio atacadista de São Paulo, SP. Foram colhidas 85 amostras de filé de atum e processadas por análises bacteriológicas e PCR. Do total, 11/85 (13% amostras foram positivas para Aeromonas spp., sendo 10/11 (90,9% confirmadas como Aeromonas hydrophila pela PCR. Para Campylobacter spp., 10/85 (11,7% foram positivas, sendo 10/10 (100% identificadas como Campylobacter jejuni pelas provas bioquímicas tradicionais e PCR ressaltando-se que duas (2/85 - 2,3% amostras de atum albergavam ambos os patógenos. Trata-se do primeiro relato no Brasil de contaminação de atum fresco por Campylobacter jejuni e Aeromonas hydrophila, indicando que este alimento ingerido in natura pode ser um veículo de transmissão de agentes patogênicos, ressaltando-se a importância de estudos adicionais que deem suporte ao controle desses microrganismos em pescado consumido cru.

  3. The genomic architecture of resistance to Campylobacter jejuni intestinal colonisation in chickens.

    Science.gov (United States)

    Psifidi, A; Fife, M; Howell, J; Matika, O; van Diemen, P M; Kuo, R; Smith, J; Hocking, P M; Salmon, N; Jones, M A; Hume, D A; Banos, G; Stevens, M P; Kaiser, P

    2016-04-18

    Campylobacter is the leading cause of foodborne diarrhoeal illness in humans and is mostly acquired from consumption or handling of contaminated poultry meat. In the absence of effective licensed vaccines and inhibitors, selection for chickens with increased resistance to Campylobacter could potentially reduce its subsequent entry into the food chain. Campylobacter intestinal colonisation levels are influenced by the host genetics of the chicken. In the present study, two chicken populations were used to investigate the genetic architecture of avian resistance to colonisation: (i) a back-cross of two White Leghorn derived inbred lines [(61 x N) x N] known to differ in resistance to Campylobacter colonisation and (ii) a 9(th) generation advanced intercross (61 x N) line. The level of colonisation with Campylobacter jejuni following experimental infection was found to be a quantitative trait. A back-cross experiment using 1,243 fully informative single nucleotide polymorphism (SNP) markers revealed quantitative trait loci (QTL) on chromosomes 7, 11 and 14. In the advanced intercross line study, the location of the QTL on chromosome 14 was confirmed and refined and two new QTLs were identified located on chromosomes 4 and 16. Pathway and re-sequencing data analysis of the genes located in the QTL candidate regions identified potential pathways, networks and candidate resistance genes. Finally, gene expression analyses were performed for some of the candidate resistance genes to support the results. Campylobacter resistance in chickens is a complex trait, possibly involving the Major Histocompatibility Complex, innate and adaptive immune responses, cadherins and other factors. Two of the QTLs for Campylobacter resistance are co-located with Salmonella resistance loci, indicating that it may be possible to breed simultaneously for enhanced resistance to both zoonoses.

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

    Science.gov (United States)

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

    2011-04-26

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

  5. The PAS Domain-Containing Protein HeuR Regulates Heme Uptake in Campylobacter jejuni

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    Jeremiah G. Johnson

    2016-11-01

    Full Text Available Campylobacter jejuni is a leading cause of bacterially derived gastroenteritis. A previous mutant screen demonstrated that the heme uptake system (Chu is required for full colonization of the chicken gastrointestinal tract. Subsequent work identified a PAS domain-containing regulator, termed HeuR, as being required for chicken colonization. Here we confirm that both the heme uptake system and HeuR are required for full chicken gastrointestinal tract colonization, with the heuR mutant being particularly affected during competition with wild-type C. jejuni. Transcriptomic analysis identified the chu genes—and those encoding other iron uptake systems—as regulatory targets of HeuR. Purified HeuR bound the chuZA promoter region in electrophoretic mobility shift assays. Consistent with a role for HeuR in chu expression, heuR mutants were unable to efficiently use heme as a source of iron under iron-limiting conditions, and mutants exhibited decreased levels of cell-associated iron by mass spectrometry. Finally, we demonstrate that an heuR mutant of C. jejuni is resistant to hydrogen peroxide and that this resistance correlates to elevated levels of catalase activity. These results indicate that HeuR directly and positively regulates iron acquisition from heme and negatively impacts catalase activity by an as yet unidentified mechanism in C. jejuni.

  6. Quantification of Campylobacter jejuni contamination on chicken carcasses in France.

    Science.gov (United States)

    Duqué, Benjamin; Daviaud, Samuel; Guillou, Sandrine; Haddad, Nabila; Membré, Jeanne-Marie

    2018-04-01

    Highly prevalent in poultry, Campylobacter is a foodborne pathogen which remains the primary cause of enteritis in humans. Several studies have determined prevalence and contamination level of this pathogen throughout the food chain. However it is generally performed in a deterministic way without considering heterogeneity of contamination level. The purpose of this study was to quantify, using probabilistic tools, the contamination level of Campylobacter spp. on chicken carcasses after air-chilling step in several slaughterhouses in France. From a dataset (530 data) containing censored data (concentration contamination level (3 log 10 or more), strengthening the probabilistic analysis and facilitating result interpretation. The sampling period and sampling area (neck/leg) had a significant effect on Campylobacter contamination level. More precisely, two "seasons" were distinguished: one from January to May, another one from June to December. During the June-to-December season, the mean Campylobacter concentration was estimated to 2.6 [2.4; 2.8] log 10 (CFU/g) and 1.8 [1.5; 2.0] log 10 (CFU/g) for neck and leg, respectively. The probability of having >1000CFU/g (higher limit of European microbial criterion) was estimated to 35.3% and 12.6%, for neck and leg, respectively. In contrast, during January-to-May season, the mean contamination level was estimated to 1.0 [0.6; 1.3] log 10 (CFU/g) and 0.6 [0.3; 0.9] log 10 (CFU/g) for neck and leg, respectively. The probability of having >1000CFU/g was estimated to 13.5% and 2.0% for neck and leg, respectively. An accurate quantification of contamination level enables industrials to better adapt their processing and hygiene practices. These results will also help in refining exposure assessment models. Copyright © 2017 Elsevier Ltd. All rights reserved.

  7. Genotypic and serotypic stability of Campylobacter jejuni strains during in vitro and in vivo passage

    DEFF Research Database (Denmark)

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

    2001-01-01

    The stability of four typing methods and the sero- and genotypic stability of three Campylobacter jejuni strains were evaluated after subculturing 50 times in triplicate and after colonising mice for up to 26 days. The employed methods were Penner heat-stable serotyping; automated ribotyping (Ribo......Printing) using HaeIII restriction enzyme; pulsed-field gel electrophoresis (PFGE) using SmaI, SalI and KpnI; and random amplified polymorphic DNA analysis (RAPD) using primers 1254, 1281 and HLWL85. No changes in any of the DNA profiles or in the reactions to heat-stable antigens were identified among...... these strains after the in vitro and in vivo passages. However, one isolate became untypeable with RAPD after passage in one of the mice. In addition, eleven other C. jejuni strains of four different serotypes were subcultured ten times to screen for instability. Neither of these showed instability using PFGE...

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

    DEFF Research Database (Denmark)

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

    2005-01-01

    Aims: To apply and evaluate LG (LPS genes) genotyping, which is a genotyping method based on a cluster of genes involved in the synthesis of surface lipopolysaccharides (LPS) in Campylobacter species, for typing of Campylobacter jejuni isolates obtained from Danish broiler chickens. Furthermore...... and LG genotyping was low when applied to poultry isolates. This is in contrast to previous studies on isolates of human origin that reported a high correlation between results obtained by the two typing methods....

  9. A charcoal- and blood-free enrichment broth for isolation and PCR detection of Campylobacter jejuni and Campylobacter coli in chicken.

    Science.gov (United States)

    Solís-Soto, Luisa Y; García, Santos; Wesley, Irene; Heredia, Norma

    2011-02-01

    The microaerophilic nature of Campylobacter and its requirement of ∼5% O(2) for growth have complicated its recovery from foods. The addition to the enrichment media of oxygen quenchers such as charcoal or blood could interfere with PCR for its detection. In this study, a two-step simple aerobic method for Campylobacter detection is proposed. A modification of the Tran blood-free enrichment broth (BFEB), in which charcoal was excluded from the medium (M-BFEB), was compared with the original formulation and other enrichment broths. Campylobacter jejuni and Campylobacter coli were screened by PCR directly from the enrichment media. Various levels of pure cultures of C. jejuni and C. coli combined with Escherichia coli were inoculated into Preston, Bolton, BFEB, and the modified BFEB (M-BFEB). In addition, Campylobacter was inoculated onto retail purchased chicken skin and recovery was quantified. Rates of recovery after 24 to 48 h of enrichment at 42 °C under aerobic incubation for BFEB and M-BFEB and microaerobic incubations for Preston and Bolton broths were determined. Overall, our results indicated that the most sensitive medium was Bolton's, followed by either BFEB or M-BFEB; the least sensitive was Preston's. M-BFEB was directly coupled to a PCR assay to detect Campylobacter, avoiding intermediate plating. Campylobacter was detected in the presence of up to 10(8) E. coli cells per ml. M-BFEB facilitated detection of both C. jejuni and C. coli artificially inoculated onto chicken skin samples. M-BFEB coupled to PCR is a rapid and attractive alternative for isolation and identification of C. coli and C. jejuni from poultry. Copyright ©, International Association for Food Protection

  10. Insights into Campylobacter jejuni colonization of the mammalian intestinal tract using a novel mouse model of infection.

    Science.gov (United States)

    Stahl, Martin; Vallance, Bruce A

    2015-01-01

    A lack of relevant disease models for Campylobacter jejuni has long been an obstacle to research into this common enteric pathogen. We recently published that mice deficient in Single IgG Interleukin-1 related receptor (SIGIRR), a repressor of MyD88-dependent innate immune signaling, were highly susceptible to enteric infection by murine bacterial pathogens. Subsequently, we successfully employed these mice as an animal model for the human pathogen C. jejuni and gained substantial new insights into infection by this pathogen. The infected mice developed significant intestinal inflammation, primarily via TLR4 stimulation. Furthermore, the resulting gastroenteritis was dependent on C. jejuni pathogenesis as bacterial strains suffering mutations in key virulence factors were attenuated in causing disease. The ability to infect SIGIRR-deficient mice with C. jejuni sheds new light onto how these bacteria colonize the mucus layer of the intestinal tract, invade epithelial cells, and raises new prospects for studying the virulence strategies and pathogenesis of C. jejuni.

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

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    Hiroshi Asakura

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

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

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

    2016-06-01

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

  13. Characterization of Escherichia coli from raw poultry in Belgium and impact on the detection of Campylobacter jejuni using Bolton broth.

    Science.gov (United States)

    Jasson, Vicky; Sampers, Imca; Botteldoorn, Nadine; López-Gálvez, Francisco; Baert, Leen; Denayer, Sarah; Rajkovic, Andreja; Habib, Ihab; De Zutter, Lieven; Debevere, Johan; Uyttendaele, Mieke

    2009-11-15

    A comparative study examining Bolton broth and Preston broth for enrichment and reliable detection of Campylobacter jejuni (both healthy and freeze stressed cells) was performed. Tested as pure cultures, Bolton broth enabled faster resuscitation and growth of C. jejuni compared to Preston broth. When C. jejuni was co-incubated with extended-spectrum-beta-lactamase (ESBL) producing Escherichia coli isolated from Belgian poultry meat preparations, the latter dominated in the Bolton enrichment broth and crowded the mCCDA plates. This resulted in the inability to recover C. jejuni by ISO 10272-1:2006 standard method. Preston broth did not support the growth of the ESBL E. coli isolates, but showed longer detection time of C. jejuni compared to Bolton broth. The use of the same antibiotic (sodium cefoperazone) in Bolton broth and in mCCDA plates may explain the problems encountered for detection of C. jejuni, as high numbers of ESBL E. coli present after enrichment in Bolton broth, also caused overgrowth and masked the few C. jejuni colonies present on the mCCDA plates. The use of Campylobacter spp. specific real-time PCR circumvented these problems and enabled rapid detection of the pathogen after 24h enrichment in both Bolton and Preston broth, for both healthy and freeze stressed cells.

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

    Science.gov (United States)

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

    2015-09-01

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

  15. A real-time PCR assay for the detection of Campylobacter jejuni in foods after enrichment culture.

    Science.gov (United States)

    Sails, Andrew D; Fox, Andrew J; Bolton, Frederick J; Wareing, David R A; Greenway, David L A

    2003-03-01

    A real-time PCR assay was developed for the quantitative detection of Campylobacter jejuni in foods after enrichment culture. The specificity of the assay for C. jejuni was demonstrated with a diverse range of Campylobacter species, related organisms, and unrelated genera. The assay had a linear range of quantification over six orders of magnitude, and the limit of detection was approximately 12 genome equivalents. The assay was used to detect C. jejuni in both naturally and artificially contaminated food samples. Ninety-seven foods, including raw poultry meat, offal, raw shellfish, and milk samples, were enriched in blood-free Campylobacter enrichment broth at 37 degrees C for 24 h, followed by 42 degrees C for 24 h. Enrichment cultures were subcultured to Campylobacter charcoal-cefoperazone-deoxycholate blood-free selective agar, and presumptive Campylobacter isolates were identified with phenotypic methods. DNA was extracted from enrichment cultures with a rapid lysis method and used as the template in the real-time PCR assay. A total of 66 samples were positive for C. jejuni by either method, with 57 samples positive for C. jejuni by subculture to selective agar medium and 63 samples positive in the real-time PCR assay. The results of both methods were concordant for 84 of the samples. The total time taken for detection from enrichment broth samples was approximately 3 h for the real-time PCR assay, with the results being available immediately at the end of PCR cycling, compared to 48 h for subculture to selective agar. This assay significantly reduces the total time taken for the detection of C. jejuni in foods and is an important model for other food-borne pathogens.

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

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    Bui Xuan

    2012-10-01

    Full Text Available Abstract 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. jejuni and on its uptake by and intracellular survival within Acanthamoeba castellanii. Results Heat, starvation and osmotic stress reduced the survival of C. jejuni significantly, whereas oxidative stress had no effect. Quantitative RT-PCR experiments showed that the transcription of virulence genes was slightly up-regulated under heat and oxidative stresses but down-regulated under starvation and osmotic stresses, the htrA gene showing the largest down-regulation in response to osmotic stress. Pre-exposure of bacteria to low nutrient or osmotic stress reduced bacterial uptake by amoeba, but no effect of heat or oxidative stress was observed. Finally, C. jejuni rapidly lost viability within amoeba cells and pre-exposure to oxidative stress had no significant effect on intracellular survival. However, the numbers of intracellular bacteria recovered 5 h post-gentamicin treatment were lower with starved, heat treated or osmotically stressed bacteria than with control bacteria. Also, while ~1.5 × 103 colony forming unit/ml internalized bacteria could typically be recovered 24 h post-gentamicin treatment with control bacteria, no starved, heat treated or osmotically stressed bacteria could be recovered at this time point. Overall, pre-exposure of C. jejuni to environmental stresses did not promote intracellular survival in A. castellanii

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

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    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.

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

    Science.gov (United States)

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

    2010-10-04

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

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

    Science.gov (United States)

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

    2015-09-02

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

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

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    Lea-Maxie Haag

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

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

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    Alexandre Thibodeau

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

  2. A Genome-Wide Association Study to Identify Diagnostic Markers for Human Pathogenic Campylobacter jejuni Strains

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    Cody J. Buchanan

    2017-06-01

    Full Text Available Campylobacter jejuni is a leading human enteric pathogen worldwide and despite an improved understanding of its biology, ecology, and epidemiology, limited tools exist for identifying strains that are likely to cause disease. In the current study, we used subtyping data in a database representing over 24,000 isolates collected through various surveillance projects in Canada to identify 166 representative genomes from prevalent C. jejuni subtypes for whole genome sequencing. The sequence data was used in a genome-wide association study (GWAS aimed at identifying accessory gene markers associated with clinically related C. jejuni subtypes. Prospective markers (n = 28 were then validated against a large number (n = 3,902 of clinically associated and non-clinically associated genomes from a variety of sources. A total of 25 genes, including six sets of genetically linked genes, were identified as robust putative diagnostic markers for clinically related C. jejuni subtypes. Although some of the genes identified in this study have been previously shown to play a role in important processes such as iron acquisition and vitamin B5 biosynthesis, others have unknown function or are unique to the current study and warrant further investigation. As few as four of these markers could be used in combination to detect up to 90% of clinically associated isolates in the validation dataset, and such markers could form the basis for a screening assay to rapidly identify strains that pose an increased risk to public health. The results of the current study are consistent with the notion that specific groups of C. jejuni strains of interest are defined by the presence of specific accessory genes.

  3. Structural and Functional Basis for Targeting Campylobacter jejuni Agmatine Deiminase To Overcome Antibiotic Resistance.

    Science.gov (United States)

    Shek, Roger; Dattmore, Devon A; Stives, Devin P; Jackson, Ashley L; Chatfield, Christa H; Hicks, Katherine A; French, Jarrod B

    2017-12-26

    Campylobacter jejuni is the most common bacterial cause of gastroenteritis and a major contributor to infant mortality in the developing world. The increasing incidence of antibiotic-resistant C. jejuni only adds to the urgency to develop effective therapies. Because of the essential role that polyamines play, particularly in protection from oxidative stress, enzymes involved in the biosynthesis of these metabolites are emerging as promising antibiotic targets. The recent description of an alternative pathway for polyamine synthesis, distinct from that in human cells, in C. jejuni suggests this pathway could be a target for novel therapies. To that end, we determined X-ray crystal structures of C. jejuni agmatine deiminase (CjADI) and demonstrated that loss of CjADI function contributes to antibiotic sensitivity, likely because of polyamine starvation. The structures provide details of key molecular features of the active site of this protein. Comparison of the unliganded structure (2.1 Å resolution) to that of the CjADI-agmatine complex (2.5 Å) reveals significant structural rearrangements that occur upon substrate binding. The shift of two helical regions of the protein and a large conformational change in a loop near the active site generate a narrow binding pocket around the bound substrate. This change optimally positions the substrate for catalysis. In addition, kinetic analysis of this enzyme demonstrates that CjADI is an iminohydrolase that effectively deiminates agmatine. Our data suggest that C. jejuni agmatine deiminase is a potentially important target for combatting antibiotic resistance, and these results provide a valuable framework for guiding future drug development.

  4. Campylobacter jejuni strains coresistant to tetracycline and ciprofloxacin in patients with gastroenteritis in Croatia.

    Science.gov (United States)

    Carev, Merica; Kovačić, Ana; Novak, Anita; Tonkić, Marija; Jerončić, Ana

    2017-04-01

    Antibiotic-resistant Campylobacter jejuni strains are rapidly emerging worldwide. Here, we aimed to determine the antibiotic-resistance patterns and genetic structure of C. jejuni from stool samples of symptomatic patients in Dalmatia, the largest Croatian county. In a population-based laboratory surveillance programme for campylobacteriosis in Dalmatia from May 2012 to May 2013, C. jejuni (n = 76) were collected from stool samples of all the patients hospitalized with gastroenteritis and matched positive outpatients (n = 77). Antibiotic susceptibility testing and pulsed-field gel electrophoresis (PFGE) genotyping of isolates were performed. Approximately 60% of the isolates were resistant to ciprofloxacin, whereas 24% of isolates were resistant to tetracycline; of the latter, 89% were also coresistant to ciprofloxacin. Resistance to erythromycin and gentamicin was infrequent (≤ 0.7%). Antibiotic-resistant strains were generally not associated with the need for hospitalization. However, the prevalence of coresistant strains increased sharply after 2010, and these coresistant strains were more prevalent in infections caused by clonal PFGE types, with distinct patterns of temporal occurrence and age distribution in infected patients. A high prevalence of coresistant Tc R /Cip R C. jejuni strains were detected in patients in Croatia. Strains were significantly associated with several clonal-type PFGE genotypes, shared common patterns of temporal occurrence, and showed distinct age distribution in infected patients, suggestive of newly identified strains. Since a high prevalence of coresistant Tc R /Cip R strains was also observed in other countries, further in-depth studies are essential to evaluate whether this phenomenon is linked to C. jejuni epidemiology in food animals and agricultural ecosystems.

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

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    Dharanesh Gangaiah

    Full Text Available BACKGROUND: Inorganic polyphosphate (poly P plays an important role in stress tolerance and virulence in many bacteria. PPK1 is the principal enzyme involved in poly P synthesis, while PPK2 uses poly P to generate GTP, a signaling molecule that serves as an alternative energy source and a precursor for various physiological processes. Campylobacter jejuni, an important cause of foodborne gastroenteritis in humans, possesses homologs of both ppk1 and ppk2. ppk1 has been previously shown to impact the pathobiology of C. jejuni. METHODOLOGY/PRINCIPAL FINDINGS: Here, we demonstrate for the first time that the deletion of ppk2 in C. jejuni resulted in a significant decrease in poly P-dependent GTP synthesis, while displaying an increased intracellular ATP:GTP ratio. The Deltappk2 mutant exhibited a significant survival defect under osmotic, nutrient, aerobic, and antimicrobial stresses and displayed an enhanced ability to form static biofilms. However, the Deltappk2 mutant was not defective in poly P and ppGpp synthesis suggesting that PPK2-mediated stress tolerance is not ppGpp-mediated. Importantly, the Deltappk2 mutant was significantly attenuated in invasion and intracellular survival within human intestinal epithelial cells as well as in chicken colonization. CONCLUSIONS/SIGNIFICANCE: Taken together, we have highlighted the role of PPK2 as a novel pathogenicity determinant that is critical for C. jejuni survival, adaptation, and persistence in the host environments. PPK2 is absent in humans and animals; therefore, can serve as a novel target for therapeutic intervention of C. jejuni infections.

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

    DEFF Research Database (Denmark)

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

    2001-01-01

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

  7. Detection of genetic diversity in Campylobacter jejuni isolated from a commercial turkey flock using flaA typing, MLST analysis and microarray assay.

    Directory of Open Access Journals (Sweden)

    Hosny El-Adawy

    Full Text Available Campylobacter is genetically highly diverse and undergoes frequent intraspecific recombination. Turkeys have been identified as an important reservoir for Campylobacter jejuni which is of public health significance. The assessment of the genetic diversity among Campylobacter population is critical for our understanding of the epidemiology of this bacterium. The genetic profiles were different according to the molecular typing methods used. The performance of established flaA genotyping, multilocus sequencing typing (MLST and DNA microarray assay based on the ArrayTube™ technology was evaluated using 14 Campylobacter jejuni isolated from a commercial turkey flock. The flaA typing was performed using PCR-RFLP with restriction enzymes Sau3AI, AluI, a 'composite' flaA analysis of AluI and Sau3AI and DdeI. The 14 isolates were differentiated into 3, 5, 7 and 9 genotypes, respectively. Entire flaA gene and short variable region (SVR sequences were analysed. Sequencing of the entire flaA provided 11 different genotypes. flaA-SVR sequence analysis detected 8 flaA alleles and 4 flaA peptides. One new flaA allele type (528 was identified. MLST analysis represented 10 different sequence types (STs and 5 clonal complexes (CCs. The microarray assay recognised 14 different genotypes. The discriminatory indices were 0.560, 0.802, 0.857, and 0.912 for flaA-RFLP depending on the used enzymes, 0.890 for flaA-SVR, 0.967 for entire flaA sequencing, 0.945 for MLST and 1.00 for the DNA microarray assay. The flaA gene was genetically stable over 20 passages on blood agar. In conclusion, the different typing tools demonstrated a high level of genetic heterogeneity of Campylobacter jejuni in a turkey flock, indicating that a single flock can be infected by multiple genotypes within one rearing cycle. DNA microarray-based assays had the highest discriminatory power when compared with other genotyping tools.

  8. The crucial role of Campylobacter jejuni genes in anti-ganglioside antibody induction in Guillain-Barré syndrome

    NARCIS (Netherlands)

    B.C. Jacobs (Bart); H.P. Endtz (Hubert); A.P. Heikema (Astrid); M. Gilbert (Michel); T. Komagamine (Tomoko); C.W. Ang (Wim); J. Glerum (Jobine); D. Brochu (Denis); J. Li (Jianjun); N. Yuki (Nobuhiro); A.F. van Belkum (Alex); P.C.R. Godschalk (Peggy)

    2004-01-01

    textabstractMolecular mimicry of Campylobacter jejuni lipo-oligosaccharides (LOS) with gangliosides in nervous tissue is considered to induce cross-reactive antibodies that lead to Guillain-Barre syndrome (GBS), an acute polyneuropathy. To determine whether specific bacterial genes

  9. RpoD promoters in Campylobacter jejuni exhibit a strong periodic signal instead of a -35 box

    DEFF Research Database (Denmark)

    Petersen, Lise; Larsen, Thomas S.; Ussery, David W.

    2003-01-01

    .jejuni genome, of which 654 were located upstream of annotated genes. A similar promoter was identified in Helicobacter pylori, a close phylogenetic relative of Campylobacter, but not in Escherichia coli, Vibrio cholerae, or six other Proteobacterial genomes, or in Staphylococcus aureus. We used upstream...

  10. Characterization of Campylobacter jejuni applying flaA short variable region sequencing, multilocus sequencing and Fourier transform infrared spectroscopy

    DEFF Research Database (Denmark)

    Josefsen, Mathilde Hartmann; Bonnichsen, Lise; Larsson, Jonas

    flaA short variable region sequencing and phenetic Fourier transform infrared (FTIR) spectroscopy was applied on a collection of 102 Campylobacter jejuni isolated from continuous sampling of organic, free range geese and chickens. FTIR has been shown to serve as a valuable tool in typing...

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

    NARCIS (Netherlands)

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

    2008-01-01

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

  12. Generation of the membrane potential and its impact on the motility, ATP production and growth in Campylobacter jejuni

    Science.gov (United States)

    The generation of an electrical membrane potential (''), the major constituent of the proton motive force (pmf) is crucial for the ATP synthesis, bacterial growth and motility. The pmf drives the rotation of flagella and is vital for the microaerophilic human pathogen Campylobacter jejuni to coloniz...

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

    NARCIS (Netherlands)

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

    2008-01-01

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

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

    Directory of Open Access Journals (Sweden)

    CORCIONIVOSCHI N.

    2007-01-01

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

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

    Science.gov (United States)

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

  16. Rapid selection of quinolone resistance in Campylobacter jejuni but not in Escherichia coli in individually housed broilers

    NARCIS (Netherlands)

    Boven, van R.M.; Veldman, K.T.; Jong, de M.C.M.; Mevius, D.J.

    2003-01-01

    Objective: To determine the within-host population dynamics of Campylobacter jejuni and Escherichia coli in chickens during and after treatment with fluoroquinolones. Materials and methods: Total and resistant faecal counts were determined from cloacal swabs during and after treatment with

  17. A Deep-Rough Mutant of Campylobacter Jejuni 81-176 Is Noninvasive for Intestinal Epithelial Cells

    Science.gov (United States)

    2004-04-01

    compared to what was seen with the wild-type strain. The food - borne pathogen Campylobacter jejuni is one of the principal causes of human gastroenteritis...point-source outbreak of campylobacteriosis associated with consumption of raw milk. J. Infect. Dis. 152:592–596. 15. Moran, A., B. Appelmelk, and G

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

    NARCIS (Netherlands)

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

    2012-01-01

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

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

    NARCIS (Netherlands)

    Ellis-Iversen, J

    2009-01-01

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

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

    DEFF Research Database (Denmark)

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

    2016-01-01

    Background: Reducing the occurrence of campylobacteriosis is a food safety issue of high priority, as in recent years it has been the most commonly reported zoonosis in the EU. Livestock farms are of particular interest, since cattle, swine and poultry are common reservoirs of Campylobacter spp....... The farm environment provides attractive foraging and breeding habitats for some bird species reported to carry thermophilic Campylobacter spp. We investigated the Campylobacter spp. carriage rates in 52 wild bird species present on 12 Danish farms, sampled during a winter and a summer season, in order...... to study the factors influencing the prevalence in wild birds according to their ecological guild. In total, 1607 individual wild bird cloacal swab samples and 386 livestock manure samples were cultured for Campylobacter spp. according to the Nordic Committee on Food Analysis method NMKL 119.Results...

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

    Science.gov (United States)

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

    2017-06-15

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

  2. Contribution of amino acid catabolism to the tissue specific persistence of Campylobacter jejuni in a murine colonization model.

    Directory of Open Access Journals (Sweden)

    Dirk Hofreuter

    Full Text Available Campylobacter jejuni is a major cause of food-borne disease in industrialized countries. Carbohydrate utilization by C. jejuni is severely restricted, and knowledge about which substrates fuel C. jejuni infection and growth is limited. Some amino acids have been shown to serve as carbon sources both in vitro and in vivo. In the present study we investigated the contribution of serine and proline catabolism to the invitro and invivo growth of C. jejuni 81-176. We confirmed that the serine transporter SdaC and the serine ammonia-lyase SdaA are required for serine utilization, and demonstrated that a predicted proline permease PutP and a bifunctional proline/delta-1-pyrroline-5-carboxylate dehydrogenase PutA are required for proline utilization by C. jejuni 81-176. C. jejuni 81-176 mutants unable to utilize serine were shown to be severely defective for colonization of the intestine and systemic tissues in a mouse model of infection. In contrast, C. jejuni 81-176 mutants unable to utilize proline were only defective for intestinal colonization. These results further emphasize the importance of amino acid utilization in C. jejuni colonization of various tissues.

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

    Science.gov (United States)

    Bui, Xuan Thanh; Winding, Anne; Qvortrup, Klaus; Wolff, Anders; Bang, Dang Duong; Creuzenet, Carole

    2012-08-01

    Campylobacter jejuni is a major cause of infectious diarrhoea worldwide but relatively little is known about its ecology. In this study, we examined its interactions with Acanthamoeba castellanii, a protozoan suspected to serve as a reservoir for bacterial pathogens. We observed rapid degradation of intracellular C.jejuni in A.castellanii 5 h post gentamicin treatment at 25°C. Conversely, we found that A.castellanii promoted the extracellular growth of C.jejuni in co-cultures at 37°C in aerobic conditions. This growth-promoting effect did not require amoebae - bacteria contact. The growth rates observed with or without contact with amoeba were similar to those observed when C.jejuni was grown in microaerophilic conditions. Preconditioned media prepared with live or dead amoebae cultivated with or without C.jejuni did not promote the growth of C.jejuni in aerobic conditions. Interestingly, the dissolved oxygen levels of co-cultures with or without amoebae - bacteria contact were much lower than those observed with culture media or with C.jejuni alone incubated in aerobic conditions, and were comparable with levels obtained after 24 h of growth of C.jejuni under microaerophilic conditions. Our studies identified the depletion of dissolved oxygen by A.castellanii as the major contributor for the observed amoeba-mediated growth enhancement. © 2011 Society for Applied Microbiology and Blackwell Publishing Ltd.

  4. Fluoroquinolone and macrolide resistance in Campylobacter jejuni isolated from broiler slaughterhouses in southern Brazil.

    Science.gov (United States)

    Sierra-Arguello, Yuli M; Perdoncini, G; Morgan, R B; Salle, C T P; Moraes, H L S; Gomes, Marcos J P; do Nascimento, Vladimir Pinheiro

    2016-01-01

    Campylobacter jejuni is recognized as a leading cause of acute bacterial gastroenteritis in humans. The over-use of antimicrobials in the human population and in animal husbandry has led to an increase in antimicrobial-resistant infections, particularly with fluoroquinolones and macrolides. The aim of the present study was to provide information of the current status of antimicrobial resistance patterns in Campylobacter jejuni from poultry sources. Fifty strains were recovered from broiler slaughterhouses in Rio Grande do Sul state, Brazil, 2012. The strains were investigated for antimicrobial susceptibility against three agents (ciprofloxacin, nalidixic acid and erythromycin) by minimal inhibitory concentrations. The strains were analysed by polymerase chain reaction-restriction fragment length polymorphism for detection of the Thr-86 mutation that confers resistance to ciprofloxacin. In addition, all the strains were tested for the presence of efflux systems (cmeB gene) conferring antimicrobial resistance. The minimum inhibitory concentrations results showed that 98% of isolates were sensitive to erythromycin and most isolates were resistant to ciprofloxacin (94%) and nalidixic acid (90%). A complete correlation was observed between the minimum inhibitory concentrations and PCR-RFLP assay. Finally, the cmeB gene that is responsible for multidrug resistance was detected in 16 isolates out the 50 strains (32%).

  5. Comparative genotyping of Campylobacter jejuni strains from patients with Guillain-Barré syndrome in Bangladesh.

    Directory of Open Access Journals (Sweden)

    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

  6. Molecular Characterization, Antimicrobial Resistance and Caco-2 Cell Invasion Potential of Campylobacter jejuni/coli from Young Children with Diarrhea.

    Science.gov (United States)

    Pan, Haijian; Ge, Yanling; Xu, Hao; Zhang, Jianmin; Kuang, Dai; Yang, Xiaowei; Su, Xudong; Huang, Zheng; Shi, Xianming; Xu, Xuebin; Meng, Jianghong

    2016-03-01

    Campylobacter is a major cause of bacterial gastroenteritis worldwide. Young children represent a particular age group affected by Campylobacter infection because of their limited diets and weak immune systems. In this study, a total of 110 Campylobacter (80 Campylobacter jejuni and 30 Campylobacter coli) isolated from children younger than 5 years of age with diarrhea in Shanghai, China in 2011 were examined for their genetic relationship and antimicrobial susceptibility. The presence of virulence genes and its association with invasion potential in Caco-2 cell were also determined. Multilocus sequence typing revealed 62 sequence types (STs) under 14 clonal complexes from C. jejuni and 15 STs under 2 clonal complexes from C. coli. High resistance rates among the 110 isolates were observed to nalidixic acid (88.2%), ciprofloxacin (87.3%) and tetracycline (87.3%), followed by ampicillin (30.9%), gentamicin (28.2%), clindamycin (21.8%), erythromycin (21.8%) and chloramphenicol (8.2%). Compared with that of C. jejuni (32.5%), a larger proportion of C. coli (83.3%) were resistant to multiple antimicrobials, including 16 isolates of ST-828 complex resistant to 6 antimicrobials: ciprofloxacin, clindamycin, erythromycin, gentamicin, nalidixic acid and tetracycline. Furthermore, 57 Campylobacter isolates were selected based on their distinct STs and the presence of virulence genes to determine their abilities to adhere to and invade Caco-2 cells. The level of invasion varied widely among isolates and had relatively weak correlation with the genotype data. Our findings provided baseline data on Campylobacter among young children. Active surveillance of Campylobacter is needed to better understand the epidemiology and antimicrobial resistance trends of this significant pathogen to help control and protect young children from such infections.

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

    Directory of Open Access Journals (Sweden)

    Dan J. Donoghue

    2017-04-01

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

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

    Energy Technology Data Exchange (ETDEWEB)

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

    2011-05-23

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

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

    Directory of Open Access Journals (Sweden)

    M John Albert

    2013-01-01

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

  10. High-Resolution Melting System to Perform Multilocus Sequence Typing of Campylobacter jejuni

    Science.gov (United States)

    Lévesque, Simon; Michaud, Sophie; Arbeit, Robert D.; Frost, Eric H.

    2011-01-01

    Multi-locus sequence typing (MLST) has emerged as the state-of-the-art method for resolving bacterial population genetics but it is expensive and time consuming. We evaluated the potential of high resolution melting (HRM) to identify known MLST alleles of Campylobacter jejuni at reduced cost and time. Each MLST locus was amplified in two or three sub fragments, which were analyzed by HRM. The approach was investigated using 47 C. jejuni isolates, previously characterized by classical MLST, representing isolates from diverse environmental, animal and clinical sources and including the six most prevalent sequence types (ST) and the most frequent alleles. HRM was then applied to a validation set of 84 additional C. jejuni isolates from chickens; 92% of the alleles were resolved in 35 hours of laboratory time and the cost of reagents per isolate was $20 compared with $100 for sequence-based typing. HRM has the potential to complement sequence-based methods for resolving SNPs and to facilitate a wide range of genotyping studies. PMID:21297862

  11. High-resolution melting system to perform multilocus sequence typing of Campylobacter jejuni.

    Directory of Open Access Journals (Sweden)

    Simon Lévesque

    Full Text Available Multi-locus sequence typing (MLST has emerged as the state-of-the-art method for resolving bacterial population genetics but it is expensive and time consuming. We evaluated the potential of high resolution melting (HRM to identify known MLST alleles of Campylobacter jejuni at reduced cost and time. Each MLST locus was amplified in two or three sub fragments, which were analyzed by HRM. The approach was investigated using 47 C. jejuni isolates, previously characterized by classical MLST, representing isolates from diverse environmental, animal and clinical sources and including the six most prevalent sequence types (ST and the most frequent alleles. HRM was then applied to a validation set of 84 additional C. jejuni isolates from chickens; 92% of the alleles were resolved in 35 hours of laboratory time and the cost of reagents per isolate was $20 compared with $100 for sequence-based typing. HRM has the potential to complement sequence-based methods for resolving SNPs and to facilitate a wide range of genotyping studies.

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

    International Nuclear Information System (INIS)

    Yakovleva, Maria E.; Moran, Anthony P.; Safina, Gulnara R.; Wadstroem, Torkel; Danielsson, Bengt

    2011-01-01

    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.

  13. CmeABC Multidrug Efflux Pump Contributes to Antibiotic Resistance and Promotes Campylobacter jejuni Survival and Multiplication in Acanthamoeba polyphaga

    Science.gov (United States)

    Vieira, Ana; Ramesh, Amritha; Seddon, Alan M.

    2017-01-01

    ABSTRACT Campylobacter jejuni is a foodborne pathogen that is recognized as the leading cause of human bacterial gastroenteritis. The widespread use of antibiotics in medicine and in animal husbandry has led to an increased incidence of antibiotic resistance in Campylobacter. In addition to a role in multidrug resistance (MDR), the Campylobacter CmeABC resistance-nodulation-division (RND)-type efflux pump may be involved in virulence. As a vehicle for pathogenic microorganisms, the protozoan Acanthamoeba is a good model for investigations of bacterial survival in the environment and the molecular mechanisms of pathogenicity. The interaction between C. jejuni 81-176 and Acanthamoeba polyphaga was investigated in this study by using a modified gentamicin protection assay. In addition, a possible role for the CmeABC MDR pump in this interaction was explored. Here we report that this MDR pump is beneficial for the intracellular survival and multiplication of C. jejuni in A. polyphaga but is dispensable for biofilm formation and motility. IMPORTANCE The endosymbiotic relationship between amoebae and microbial pathogens may contribute to persistence and spreading of the latter in the environment, which has significant implications for human health. In this study, we found that Campylobacter jejuni was able to survive and to multiply inside Acanthamoeba polyphaga; since these microorganisms can coexist in the same environment (e.g., on poultry farms), the latter may increase the risk of infection with Campylobacter. Our data suggest that, in addition to its role in antibiotic resistance, the CmeABC MDR efflux pump plays a role in bacterial survival within amoebae. Furthermore, we demonstrated synergistic effects of the CmeABC MDR efflux pump and TetO on bacterial resistance to tetracycline. Due to its role in both the antibiotic resistance and the virulence of C. jejuni, the CmeABC MDR efflux pump could be considered a good target for the development of antibacterial

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

    Science.gov (United States)

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

    2017-06-19

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

  15. Prevalence of Genetic Determinants and Phenotypic Resistance to Ciprofloxacin in Campylobacter jejuni from Lithuania

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    Jurgita Aksomaitiene

    2018-02-01

    Full Text Available Recently, the number of reports on isolation of ciprofloxacin resistant Campylobacter jejuni has increased worldwide. The aim of this study was to determine the prevalence of resistance to ciprofloxacin and its genetic determinants among C. jejuni isolated from humans (n = 100, poultry products (n = 96 and wild birds (n = 96 in Lithuania. 91.4% of the C. jejuni isolates were phenotypically resistant to ciprofloxacin. DNA sequence analyses of the gyrA gene from 292 isolates revealed that a change in amino acid sequence, Thr86Ile, was the main substition conferring resistance to ciprofloxacin. This change was significantly associated with isolates from poultry products (P < 0.05 and humans (P < 0.05. A total of 26.7% of C. jejuni isolates from human (n = 47, poultry products (n = 30 and wild bird (n = 1, had a mutation from Ser at position 22, and six had an additional mutation from Ala at position 39. Eight isolates from poultry and two isolates from human, corresponding to 67.0% of isolates with MICs ≥128 μg/ml, showed missense mutations Thr86Ile (ACA → ATA and Ser22Gly (AGT → GGT together, whereas isolates without these mutations showed lower MIC values (from 4 to 64 μg/ml. Two hundred forty-five C. jejuni isolates showed one or more silent mutations, and 32.4% of examined isolates possessed six silent mutations. In addition to the ciprofloxacin resistant isolates harboring only Thr86Ile point mutation (110 isolates, the current study identified resistant isolates (n = 101 harboring additional point mutations (Ser22Gly, Ala39Ser, Arg48Lys, Thr85Ala Ala122Ser, Glu136Asp, Vall49Ile, and strains (n = 57 having only Glu136Asp point mutation. The study highlight the potential public health problem with elevated ciprofloxacin resistance in Campylobacters from poultry meat, wild birds and humans, and the need for extensive surveillance enabling to follow changes of antimicrobial resistance development in this species.

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

    Science.gov (United States)

    Crespo, M D; Altermann, E; Olson, J; Miller, W G; Chandrashekhar, K; Kathariou, S

    2016-07-01

    In Campylobacter spp., resistance to the antimicrobials 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; 44,095nt) harboring tet(O) was identified in C. jejuni strain 11601MD, which was isolated from the jejunum of a turkey produced conventionally in North Carolina. Analysis of the p11601MD sequence revealed the presence of a high-GC content cassette with four genes that included tet(O) and a putative aminoglycoside transferase gene (aphA-3) highly similar to kanamycin resistance determinants. Several genes putatively involved in conjugative transfer were also identified on the plasmid. These findings will contribute to a better understanding of the distribution of potentially self-mobilizing plasmids harboring antibiotic resistance determinants in Campylobacter spp. from turkeys and other sources. Copyright © 2016 Elsevier Inc. All rights reserved.

  17. A novel Campylobacter jejuni sequence type from a culture-negative patient in the Gambia.

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    Gerard A J Morris

    2008-03-01

    Full Text Available The introduction of molecular diagnostic methods is crucial for improved understanding of the aetiology and epidemiology of bacterial infections in communities in resource poor settings. A blood sample from a 7 month old patient diagnosed with malaria in 2001 in a Gambian outpatient clinic was reported as culture negative after it was subjected to traditional bacterial culture protocols. We re-addressed the analysis of the blood sample from this case more recently (after 6.5 years in archival storage in pilot work establishing 16S rRNA PCR in our molecular laboratory. Initial 16S rRNA PCR results confirmed the presence of bacterial DNA in the sample. 16S rRNA sequence analysis identified the organism as Campylobacter spp. In light of the molecular evidence we successfully grew the organism using appropriate culture conditions and subsequently biochemically confirmed that the isolate was Campylobacter jejuni. PCR and DNA sequencing of a set of seven C. jejuni housekeeping genes and in silico Multilocus Sequence Typing (MLST analysis revealed that the isolate exhibits a novel sequence type (ST of C. jejuni (ST 2928 and belongs to ST-443 clonal complex. This study demonstrates the potential for molecular tools to enhance the diagnosis of bacterial infections, which remain a major killer globally, not least in children in the developing world. Improvements in diagnostics are needed, and will be important not only for sick individuals but also for populations, where better measures of disease burden will contribute significantly to the improvement of public health policy.

  18. Detection and genotypic differentiation of Campylobacter jejuni and Campylobacter coli strains from laying hens by multiplex PCR and fla-typing.

    Science.gov (United States)

    Müller, Wolfgang; Böhland, Corinna; Methner, Ulrich

    2011-12-01

    In total, 26 Campylobacter (C.) strains, isolated from liver, spleen, caecal or jejunal content of laying hens from different flocks were examined. In these flocks a drop in egg production, an increasing mortality and livers with whitish-grey lesions as post-mortem finding were observed. Suspected Campylobacter colonies were differentiated using a modified m-PCR in 13 Campylobacter jejuni and 13 Campylobacter coli strains. All isolates were characterised by typing of the flaA and flaB gene each with two restriction enzymes. To compare the four different profiles for all strains an artificial "fla-type" was generated. Different and identical fla-types of C. jejuni and C. coli were recovered from both intestinal and extra-intestinal organs of the laying hens and even from individual birds. One significant observation is that some fla-types of C. jejuni or C. coli were detected in intestinal and systemic sites but not all fla-types of both species appeared to be equally able to invade internal organs. Copyright © 2011 Elsevier Ltd. All rights reserved.

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

    Energy Technology Data Exchange (ETDEWEB)

    Song, Wan Seok; Nam, Mi Sun; Namgung, Byeol [Department of Systems Immunology, College of Biomedical Science, Kangwon National University, Chuncheon 200-701 (Korea, Republic of); Yoon, Sung-il, E-mail: sungil@kangwon.ac.kr [Department of Systems Immunology, College of Biomedical Science, Kangwon National University, Chuncheon 200-701 (Korea, Republic of); Institute of Bioscience and Biotechnology, Kangwon National University, Chuncheon 200-701 (Korea, Republic of)

    2015-03-20

    Campylobacter jejuni is a bacterium that uses flagella for motility and causes worldwide acute gastroenteritis in humans. The C. jejuni N-acetyltransferase PseH (cjPseH) is responsible for the third step in flagellin O-linked glycosylation and plays a key role in flagellar formation and motility. cjPseH transfers an acetyl group from an acetyl donor, acetyl coenzyme A (AcCoA), to the amino group of UDP-4-amino-4,6-dideoxy-N-acetyl-β-L-altrosamine to produce UDP-2,4-diacetamido-2,4,6-trideoxy-β-L-altropyranose. To elucidate the catalytic mechanism of cjPseH, crystal structures of cjPseH alone and in complex with AcCoA were determined at 1.95 Å resolution. cjPseH folds into a single-domain structure of a central β-sheet decorated by four α-helices with two continuously connected grooves. A deep groove (groove-A) accommodates the AcCoA molecule. Interestingly, the acetyl end of AcCoA points toward an open space in a neighboring shallow groove (groove-S), which is occupied by extra electron density that potentially serves as a pseudosubstrate, suggesting that the groove-S may provide a substrate-binding site. Structure-based comparative analysis suggests that cjPseH utilizes a unique catalytic mechanism of acetylation that has not been observed in other glycosylation-associated acetyltransferases. Thus, our studies on cjPseH will provide valuable information for the design of new antibiotics to treat C. jejuni-induced gastroenteritis. - Highlights: • cjPseH adopts a single-domain structure of a central β-sheet decorated by α-helices. • cjPseH features two continuously connected grooves on the protein surface. • Acetyl coenzyme A (AcCoA) binds into a deep groove of cjPseH in an ‘L’ shape. • The acetyl end of AcCoA points to a wide groove, a potential substrate-binding site.

  20. Comparison of real-time PCR assays for detection, quantification, and differentiation of campylobacter jejuni and campylobacter coli in broiler neck skin samples.

    Science.gov (United States)

    Schnider, A; Overesch, G; Korczak, B M; Kuhnert, P

    2010-06-01

    We tested the use of multiplex real-time PCR for detection and quantification of Campylobacter jejuni and Campylobacter coli on broiler carcass neck skin samples collected during 2008 from slaughterhouses in Switzerland. Results from an established TaqMan assay based on two different targets (hipO and ceuE for C. jejuni and C. coli, respectively) were corroborated with data from a newly developed assay based on a single-nucleotide polymorphism in the fusA gene, which allows differentiation between C. jejuni and C. coli. Both multiplex real-time PCRs were applied simultaneously for direct detection, differentiation, and quantification of Campylobacter from 351 neck skin samples and compared with culture methods. There was good correlation in detection and enumeration between real-time PCR results and quantitative culture, with real-time PCR being more sensitive. Overall, 251 (71.5%) of the samples were PCR positive for Campylobacter, with 211 (60.1%) in the hipO-ceuE assays, 244 (69.5%) in the fusA assay, and 204 (58.1%) of them being positive in both PCR assays. Thus, the fusA assay was similarly sensitive to the enrichment culture (72.4% positive); however, it is faster and allows for quantification. In addition, real-time PCR allowed for species differentiation; roughly 60% of positive samples contained C. jejuni, less than 10% C. coli, and more than 30% contained both species. Real-time PCR proved to be a suitable method for direct detection, quantification, and differentiation of Campylobacter from carcasses, and could permit time-efficient surveillance of these zoonotic agents.

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

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    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

  2. Comparing the Genetic Diversity and Antimicrobial Resistance Profiles of Campylobacter jejuni Recovered from Cattle and Humans

    Directory of Open Access Journals (Sweden)

    Wonhee Cha

    2017-05-01

    Full Text Available Campylobacter jejuni, a leading cause of gastroenteritis in humans, is a foodborne pathogen that can reside in chickens, pigs, and cattle. Because resistance to fluoroquinolones and macrolides, which are commonly used to treat human infections, has emerged in C. jejuni, it is imperative to continously monitor resistance patterns and examine the genetic variation in strains from human infections and animal reservoirs. Our previous study of C. jejuni from human campylobacteriosis cases showed a significantly higher rate of tetracycline resistance compared to national trends, and identified multilocus sequence type (ST-982 and a history of cattle contact to be associated with tetracycline resistance. To further investigate these associations, we conducted a cross-sectional study to determine the frequency of antimicrobial resistance and examine the genetic diversity of C. jejuni recovered from 214 cattle at three Michigan herds. Overall, the prevalence of C. jejuni was 69.2% (range: 58.6–83.8% for the three farms, and 83.7% (n = 113 of isolates were resistant to one or more antimicrobials. Resistance to only tetracycline predominated among the cattle isolates (n = 89; 65.9% with most resistant strains belonging to ST-459 (96.5% or ST-982 (86.4%. Among the 22 STs identified, STs 459 and 982 were more prevalent in one feedlot, which reported the use of chlortetracycline in feed upon arrival of a new herd. PCR-based fingerprinting demonstrated that the ST-982 isolates from cattle and humans had identical banding patterns, suggesting the possibility of interspecies transmission. Resistance to macrolides (1.5% and ciprofloxacin (16.3% was also observed; 14 of the 22 ciprofloxacin resistant isolates represented ST-1244. Together, these findings demonstrate a high prevalence of antimicrobial resistant C. jejuni in cattle and identify associations with specific genotypes. Continuous monitoring and identification of risk factors for resistance emergence

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

    DEFF Research Database (Denmark)

    Gill, Carson; Bahrndorff, Simon; Lowenberger, Carl

    2016-01-01

    The house fly, Musca domestica, has been implicated as a vector of Campylobacter spp., a major cause of human disease. Little is known whether house flies serve as biological amplifying hosts or mechanical vectors for Campylobacter jejuni. We investigated the period after C. jejuni had been...... after ingestion but no countable C. jejuni colonies were observed > 24 hours post-ingestion. We detected viable C. jejuni in house fly vomitus and excreta up to 4 hours after ingestion, but no viable bacteria were detected ≥ 8 hours. Suppression subtractive hybridization identified pathogen-induced gene...... quantitative real time PCR. Some house fly factor, or combination of factors, eliminates C. jejuni within 24 hours post-ingestion. Because C. jejuni is not amplified within the body of the housefly, this insect likely serves as a mechanical vector rather than as a true biological, amplifying vector for C...

  4. Detection of Campylobacter jejuni in dairy farm environmental samples using SYBR Green real-time polymerase chain reaction.

    Science.gov (United States)

    Nam, H M; Srinivasan, V; Murinda, S E; Oliver, S P

    2005-01-01

    The aim of this study was to evaluate a SYBR Green based real-time PCR assay using well-characterized primers to detect Campylobacter jejuni in naturally contaminated dairy farm environmental samples. Specificity of the assay was determined with 62 C. jejuni strains and 120 non-C. jejuni strains. Peak melting temperature obtained with melting curves specific for C. jejuni was 77.5 degrees C. Standard curves were constructed using mean threshold cycle (C(T)) and various concentrations of C. jejuni ranging from 10(0) to 10(8) colony forming units (CFU)/mL, which resulted in a linear relationship between C(T) and log input DNA. Correlation coefficients of standard curves based on pure culture of C. jejuni in broth and spiked cells in lagoon water were R(2) = 0.995 (slope = 3.21) and R(2) = 0.988 (slope = 3.22), respectively, and sensitivity limits were 10(3) CFU/mL, respectively. After 24-h enrichment, total C. jejuni counts of all samples spiked with 10(0) CFU/mL reached >10(5) CFU/mL, and the detection limit was improved from >10(3) CFU/mL to PCR assay detected C. jejuni in 25 (30.4%) of 82 samples, with 17 (68%) of these samples being culture positive for C. jejuni. All samples that were positive by standard culture methods were also positive by the real-time PCR method. Mean C( T ) values of 48-h enriched cultures for 17 PCR-positive/culture-positive samples and eight PCR-positive/culture-negative samples were 21.4 +/- 3.6, and 34.6 +/- 1.5 (p 38.0. These results indicate that the SYBR Green real-time PCR assay provides a specific, reproducible, and simple method for detecting C. jejuni in dairy farm environmental samples.

  5. Detection of Campylobacter jejuni and Campylobacter coli from broiler chicken-related samples using BAX PCR and conventional International Organization for Standardization culture.

    Science.gov (United States)

    Williams, Lisa K; McMeechan, Alisdair; Baalham, Tamsin; Ward, Laura; Humphrey, Tom J; Jørgensen, Frieda

    2008-04-01

    In this study, the conventional International Organization for Standardization (ISO) culture method was compared with the DuPont Qualicon BAX system, a high-throughput, rapid molecular assay that can be used to detect several bacterial species, including Campylobacter jejuni and Campylobacter coli in diverse sample types. Standard enrichment culture is a time-consuming process, taking up to 6 days to obtain a confirmed result. Rapid molecular assays have been developed that provide results within 24 h. Naturally contaminated samples from the poultry production chain were examined for the presence of Campylobacter spp. Samples from broiler chicken ceca (n = 100), fresh chicken carcass rinses (n = 60), and bootsocks (gauze sock walked through a broiler chicken house; n = 50) were enriched according to the ISO 10272 method in Bolton broth specifically designed to detect Campylobacter spp. in complex sample types. Samples were enriched without blood for use with the BAX system using the Campylobacter BAX kits for the detection of C. jejuni and C. coli. Samples also were directly plated onto modified charcoal cefperazone deoxycholate agar, and results were compared with those from the enriched samples for the ability to detect Campylobacter spp. Campylobacter spp. were isolated from 49% of samples with conventional enrichment cultures, from 48% with direct culture, from 68% with the BAX system and enrichment cultures, and from 62% with the BAX system used directly with samples. Overall, the BAX system detected more positive samples than did the conventional culture method and is an effective methodology for the rapid and reliable detection of Campylobacter spp. from diverse sample types.

  6. Important role of a putative lytic transglycosylase Cj0843c in β-lactam resistance in Campylobacter jejuni

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    Ximin eZeng

    2015-11-01

    Full Text Available Beta-lactam antibiotics are an important class of antibiotics for treating bacterial infections. Despite prevalent β-lactam resistance in Campylobacter jejuni, the leading bacterial cause of human diarrhea in developed countries, molecular mechanism of β-lactam resistance in C. jejuni is still largely unknown. In this study, C. jejuni 81-176 was used for random transposon mutagenesis. Screening of a 2,800-mutant library identified 22 mutants with increased susceptibility to ampicillin. Of these mutants, two mutants contains mutations in Cj0843c (a putative lytic transglycosylase gene and in its upstream gene Cj0844c, respectively. Molecular manipulation in different strains demonstrated that Cj0843c contributes to both intrinsic and acquired β-lactam resistance in C. jejuni. Consistent with this finding, inactivation of Cj0843c also dramatically reduced β-lactamase activity. Genomic examination and PCR analysis showed Cj0843c is widely distributed in C. jejuni. The Cj0843 was localized in the periplasm as demonstrated by immunoblotting using specific antibodies. Turbidimetric assay further demonstrated the capability of the purified Cj0843c to hydrolyze cell walls. Inactivation of Cj0843c also significantly reduced C. jejuni colonization in the intestine. Together, this study identifies a mechanism of β-lactam resistance in C. jejuni and provides insights into the role of cell wall metabolism in regulating β-lactamase activity.

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

    DEFF Research Database (Denmark)

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

    2016-01-01

    Background: Reducing the occurrence of campylobacteriosis is a food safety issue of high priority, as in recent years it has been the most commonly reported zoonosis in the EU. Livestock farms are of particular interest, since cattle, swine and poultry are common reservoirs of Campylobacter spp. ...

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

    NARCIS (Netherlands)

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

    2016-01-01

    Campylobacter is the most common causative agent of human bacterial gastroenteritis and is frequently found in surface water, where it indicates recent contamination with animal faeces, sewage effluent, and agricultural run-off. The contribution of different animal reservoirs to surface water

  9. Correlation between genotypic diversity, lipooligosaccharide gene locus class variation and Caco-2 invasion potential of Campylobacter jejuni from human and chicken meat origin: a contribution to virulotyping

    Science.gov (United States)

    Significant interest in studying the lipooligosaccharide (LOS) of Campylobacter jejuni stemmed from its potential role in post-infection paralytic disorders. In this study we present PCR screening of five LOS locus classes (A, B, C, D, and E), for a collection of 117 C. jejuni strains from chicken m...

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

    DEFF Research Database (Denmark)

    Li, Yiping; Ingmer, Hanne; Madsen, Mogens

    2008-01-01

    Background Campylobacter jejuni is a major cause of inflammatory diarrhoea in humans and is considered a commensal of the gastroenteric tract of the avian host. However, little is known about the interaction between C. jejuni and the avian host including the cytokine responses and the expression...

  11. Detection of Campylobacter jejuni by culture and real-time PCR in a French cohort of patients with Guillain-Barre syndrome.

    Science.gov (United States)

    Sivadon-Tardy, Valérie; Orlikowski, David; Porcher, Raphael; Ronco, Esthel; Caudie, Christiane; Roussi, Jacqueline; Fauchère, Jean-Louis; Mégraud, Francis; Tabor, Helen; Sharshar, Tarek; Annane, Djillali; Raphaël, Jean-Claude; Gaillard, Jean-Louis

    2010-06-01

    Bacteriological culture and real-time PCR (RT-PCR) were used to detect Campylobacter jejuni in fecal samples from a French cohort of 237 patients with Guillain-Barré syndrome (GBS). We provide evidence that diverse serotypes and genotypes of C. jejuni are a major trigger of GBS in France.

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

    Directory of Open Access Journals (Sweden)

    A.F. Carvalho

    2010-10-01

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

  13. Campylobacter-Associated Diseases in Animals.

    Science.gov (United States)

    Sahin, Orhan; Yaeger, Michael; Wu, Zuowei; Zhang, Qijing

    2017-02-08

    Campylobacter includes a group of genetically diverse species causing a range of diseases in animals and humans. The bacterium is frequently associated with two economically important and epidemiologically distinct reproductive diseases in ruminants: enzootic infectious infertility in cattle owing to Campylobacter fetus subsp. venerealis and abortions in sheep, goats, and cattle. Septic abortion, usually epizootic in sheep, has been historically associated with C. fetus subsp. fetus and to a lesser extent with Campylobacter jejuni. However, there has been a dramatic species shift in the etiology of Campylobacter abortions in recent years: C. jejuni has now replaced C. fetus subsp. fetus as the predominant cause of sheep abortion in the United States, which appears to be driven primarily by clonal expansion of a hypervirulent tetracycline-resistant C. jejuni clone. Here we provide a review on the recent advances in understanding the pathobiology of Campylobacter infections in animals, with an emphasis on the diseases in ruminants, covering epidemiology, pathogenesis, genomics, and control measures.

  14. A Mutator Phenotype Promoting the Emergence of Spontaneous Oxidative Stress-Resistant Mutants in Campylobacter jejuni.

    Science.gov (United States)

    Dai, Lei; Sahin, Orhan; Tang, Yizhi; Zhang, Qijing

    2017-12-15

    Campylobacter jejuni is a leading cause of foodborne illnesses worldwide. As a microaerophilic organism, C. jejuni must be able to defend against oxidative stress encountered both in the host and in the environment. How Campylobacter utilizes a mutation-based mechanism for adaptation to oxidative stress is still unknown. Here we present a previously undescribed phenotypic and genetic mechanism that promotes the emergence of oxidative stress-resistant mutants. Specifically, we showed that a naturally occurring mutator phenotype, resulting from a loss of function mutation in the DNA repair enzyme MutY, increased oxidative stress resistance (OX R ) in C. jejuni We further demonstrated that MutY malfunction did not directly contribute to the OX R phenotype but increased the spontaneous mutation rate in the peroxide regulator gene perR , which functions as a repressor for multiple genes involved in oxidative stress resistance. Mutations in PerR resulted in loss of its DNA binding function and derepression of PerR-controlled oxidative stress defense genes, thereby conferring an OX R phenotype and facilitating Campylobacter survival under oxidative stress. These findings reveal a new mechanism that promotes the emergence of spontaneous OX R mutants in bacterial organisms. IMPORTANCE Although a mutator phenotype has been shown to promote antibiotic resistance in many bacterial species, little is known about its contribution to the emergence of OX R mutants. This work describes the link between a mutator phenotype and the enhanced emergence of OX R mutants as well as its underlying mechanism involving DNA repair and mutations in PerR. Since DNA repair systems and PerR are well conserved in many bacterial species, especially in Gram positives, the same mechanism may operate in multiple bacterial species. Additionally, we developed a novel method that allows for rapid quantification of spontaneous OX R mutants in a bacterial population. This method represents a technical

  15. MAMA-PCR assay for the detection of point mutations associated with high-level erythromycin resistance in Campylobacter jejuni and Campylobacter coli strains.

    Science.gov (United States)

    Alonso, Rodrigo; Mateo, Estibaliz; Churruca, Estibaliz; Martinez, Irati; Girbau, Cecilia; Fernández-Astorga, Aurora

    2005-10-01

    Twenty Campylobacter jejuni and 16 Campylobacter coli strains isolated from humans and food/animals, including 17 isolates resistant to erythromycin, were analyzed. A combined mismatch amplification mutation assay-PCR technique was developed to detect the mutations A 2074 C and A 2075 G in the 23S rRNA gene associated with erythromycin resistance. All high-level erythromycin-resistant strains examined by DNA sequencing carried the transition mutation A 2075 G, whereas no isolate carried the A 2074 C mutation. No mutations were found among the susceptible and low-level erythromycin-resistant strains.

  16. Campylobacter and Toll-like receptors : implications for vaccine development

    NARCIS (Netherlands)

    de Zoete, M.R.

    2010-01-01

    Campylobacter jejuni is a Gram-negative highly motile bacterium that colonizes the intestinal tract of humans, leading to inflammation of the intestinal mucosal layer. Campylobacter-induced enteritis causes (bloody) diarrhea, cramps, malaise and fever, which resolves within two weeks. In a small

  17. Phage exposure causes dynamic shifts in the expression states of specific phase-variable genes of Campylobacter jejuni

    DEFF Research Database (Denmark)

    Aidley, Jack; Holst Sørensen, Martine C.; Bayliss, Christopher D.

    2017-01-01

    Phase variation (PV) creates phenotypic heterogeneity at high frequencies and in a reversible manner. This phenomenon allows bacteria to adapt to a variety of different environments and selective pressures. In Campylobacter jejuni this reversible adaptive process is mediated by mutations in homop......Phase variation (PV) creates phenotypic heterogeneity at high frequencies and in a reversible manner. This phenomenon allows bacteria to adapt to a variety of different environments and selective pressures. In Campylobacter jejuni this reversible adaptive process is mediated by mutations...... in homopolymeric G/C tracts. Many C. jejuni-specific phages are dependent on phase-variable surface structures for successful infection. We previously identified the capsular polysaccharide (CPS) moiety, MeOPN-GalfNAc, as a receptor for phage F336 and showed that phase-variable expression of the transferase...... for this CPS modification, cj1421, and two other phase-variable CPS genes generated phage resistance in C. jejuni. Here we investigate the population dynamics of C. jejuni NCTC11168 when exposed to phage F336 in vitro using a newly described method - the 28-locus-CJ11168 PV analysis. Dynamic switching...

  18. Prevalence and Antibiotic Resistance against Tetracycline in Campylobacter jejuni and C. coli in Cattle and Beef Meat from Selangor, Malaysia

    Science.gov (United States)

    Premarathne, Jayasekara M. K. J. K.; Anuar, Aimi S.; Thung, Tze Young; Satharasinghe, Dilan A.; Jambari, Nuzul Noorahya; Abdul-Mutalib, Noor-Azira; Huat, John Tang Yew; Basri, Dayang F.; Rukayadi, Yaya; Nakaguchi, Yoshitsugu; Nishibuchi, Mitsuaki; Radu, Son

    2017-01-01

    Campylobacter is a major foodborne pathogen frequently associated with human bacterial gastroenteritis in the world. This study was conducted to determine the prevalence and antibiotic resistance of Campylobacter spp. in the beef food system in Malaysia. A total of 340 samples consisting of cattle feces (n = 100), beef (n = 120) from wet markets and beef (n = 120) from hypermarkets were analyzed for Campylobacter spp. The overall prevalence of Campylobacter was 17.4%, consisting of 33% in cattle fecal samples, 14.2% in raw beef from wet market and 7.5% in raw beef from the hypermarket. The multiplex-polymerase chain reaction (PCR) identified 55% of the strains as C. jejuni, 26% as C. coli, and 19% as other Campylobacter spp. A high percentage of Campylobacter spp. were resistant to tetracycline (76.9%) and ampicillin (69.2%), whilst low resistance was exhibited to chloramphenicol (7.6%). The MAR Index of Campylobacter isolates from this study ranged from 0.09 to 0.73. The present study indicates the potential public health risk associated with the beef food system, hence stringent surveillance, regulatory measures, and appropriate interventions are required to minimize Campylobacter contamination and prudent antibiotic usage that can ensure consumer safety. PMID:29255448

  19. Prevalence and Antibiotic Resistance against Tetracycline in Campylobacter jejuni and C. coli in Cattle and Beef Meat from Selangor, Malaysia

    Directory of Open Access Journals (Sweden)

    Jayasekara M. K. J. K. Premarathne

    2017-12-01

    Full Text Available Campylobacter is a major foodborne pathogen frequently associated with human bacterial gastroenteritis in the world. This study was conducted to determine the prevalence and antibiotic resistance of Campylobacter spp. in the beef food system in Malaysia. A total of 340 samples consisting of cattle feces (n = 100, beef (n = 120 from wet markets and beef (n = 120 from hypermarkets were analyzed for Campylobacter spp. The overall prevalence of Campylobacter was 17.4%, consisting of 33% in cattle fecal samples, 14.2% in raw beef from wet market and 7.5% in raw beef from the hypermarket. The multiplex-polymerase chain reaction (PCR identified 55% of the strains as C. jejuni, 26% as C. coli, and 19% as other Campylobacter spp. A high percentage of Campylobacter spp. were resistant to tetracycline (76.9% and ampicillin (69.2%, whilst low resistance was exhibited to chloramphenicol (7.6%. The MAR Index of Campylobacter isolates from this study ranged from 0.09 to 0.73. The present study indicates the potential public health risk associated with the beef food system, hence stringent surveillance, regulatory measures, and appropriate interventions are required to minimize Campylobacter contamination and prudent antibiotic usage that can ensure consumer safety.

  20. Genome-wide fitness analyses of the foodborne pathogen Campylobacter jejuni in in vitro and in vivo models

    DEFF Research Database (Denmark)

    de Vries, Stefan P. W.; Gupta, Srishti; Baig, Abiyad

    2017-01-01

    Campylobacter is the most common cause of foodborne bacterial illness worldwide. Faecal contamination of meat, especially chicken, during processing represents a key route of transmission to humans. There is a lack of insight into the mechanisms driving C. jejuni growth and survival within hosts......-rich and -poor conditions at 4 degrees C and infection of human gut epithelial cells was assessed by Tn-insertion site sequencing (Tn-seq). A total of 331 homologous gene clusters were essential for fitness during in vitro growth in three C. jejuni strains, revealing that a large part of its genome is dedicated...

  1. High-resolution transcriptome maps reveal strain-specific regulatory features of multiple Campylobacter jejuni isolates.

    Directory of Open Access Journals (Sweden)

    Gaurav Dugar

    2013-05-01

    Full Text Available Campylobacter jejuni is currently the leading cause of bacterial gastroenteritis in humans. Comparison of multiple Campylobacter strains revealed a high genetic and phenotypic diversity. However, little is known about differences in transcriptome organization, gene expression, and small RNA (sRNA repertoires. Here we present the first comparative primary transcriptome analysis based on the differential RNA-seq (dRNA-seq of four C. jejuni isolates. Our approach includes a novel, generic method for the automated annotation of transcriptional start sites (TSS, which allowed us to provide genome-wide promoter maps in the analyzed strains. These global TSS maps are refined through the integration of a SuperGenome approach that allows for a comparative TSS annotation by mapping RNA-seq data of multiple strains into a common coordinate system derived from a whole-genome alignment. Considering the steadily increasing amount of RNA-seq studies, our automated TSS annotation will not only facilitate transcriptome annotation for a wider range of pro- and eukaryotes but can also be adapted for the analysis among different growth or stress conditions. Our comparative dRNA-seq analysis revealed conservation of most TSS, but also single-nucleotide-polymorphisms (SNP in promoter regions, which lead to strain-specific transcriptional output. Furthermore, we identified strain-specific sRNA repertoires that could contribute to differential gene regulation among strains. In addition, we identified a novel minimal CRISPR-system in Campylobacter of the type-II CRISPR subtype, which relies on the host factor RNase III and a trans-encoded sRNA for maturation of crRNAs. This minimal system of Campylobacter, which seems active in only some strains, employs a unique maturation pathway, since the crRNAs are transcribed from individual promoters in the upstream repeats and thereby minimize the requirements for the maturation machinery. Overall, our study provides new

  2. High-Resolution Transcriptome Maps Reveal Strain-Specific Regulatory Features of Multiple Campylobacter jejuni Isolates

    Science.gov (United States)

    Förstner, Konrad U.; Heidrich, Nadja; Reinhardt, Richard; Nieselt, Kay; Sharma, Cynthia M.

    2013-01-01

    Campylobacter jejuni is currently the leading cause of bacterial gastroenteritis in humans. Comparison of multiple Campylobacter strains revealed a high genetic and phenotypic diversity. However, little is known about differences in transcriptome organization, gene expression, and small RNA (sRNA) repertoires. Here we present the first comparative primary transcriptome analysis based on the differential RNA–seq (dRNA–seq) of four C. jejuni isolates. Our approach includes a novel, generic method for the automated annotation of transcriptional start sites (TSS), which allowed us to provide genome-wide promoter maps in the analyzed strains. These global TSS maps are refined through the integration of a SuperGenome approach that allows for a comparative TSS annotation by mapping RNA–seq data of multiple strains into a common coordinate system derived from a whole-genome alignment. Considering the steadily increasing amount of RNA–seq studies, our automated TSS annotation will not only facilitate transcriptome annotation for a wider range of pro- and eukaryotes but can also be adapted for the analysis among different growth or stress conditions. Our comparative dRNA–seq analysis revealed conservation of most TSS, but also single-nucleotide-polymorphisms (SNP) in promoter regions, which lead to strain-specific transcriptional output. Furthermore, we identified strain-specific sRNA repertoires that could contribute to differential gene regulation among strains. In addition, we identified a novel minimal CRISPR-system in Campylobacter of the type-II CRISPR subtype, which relies on the host factor RNase III and a trans-encoded sRNA for maturation of crRNAs. This minimal system of Campylobacter, which seems active in only some strains, employs a unique maturation pathway, since the crRNAs are transcribed from individual promoters in the upstream repeats and thereby minimize the requirements for the maturation machinery. Overall, our study provides new insights into

  3. Campylobacter jejuni strain CG8421: a refined model for the study of Campylobacteriosis and evaluation of Campylobacter vaccines in human subjects.

    Science.gov (United States)

    Tribble, David R; Baqar, Shahida; Carmolli, Marya P; Porter, Chad; Pierce, Kristen K; Sadigh, Katrin; Guerry, Patricia; Larsson, Catherine J; Rockabrand, David; Ventone, Cassandra H; Poly, Frederic; Lyon, Caroline E; Dakdouk, Sandra; Fingar, Ann; Gilliland, Theron; Daunais, Patrick; Jones, Erika; Rymarchyk, Stacia; Huston, Christopher; Darsley, Michael; Kirkpatrick, Beth D

    2009-11-15

    A robust human challenge model for Campylobacter jejuni is an important tool for the evaluation of candidate vaccines. The previously established model conveys a potential risk of Guillain-Barré syndrome attributable to lipooligosaccharide ganglioside mimicry. This work establishes a new C. jejuni human challenge model that uses a strain (CG8421) without ganglioside mimicry and that applies Campylobacter-specific cellular immunity screening to achieve high attack rates at lower inoculum doses. Healthy Campylobacter-naive adults participated in an open-label challenge trial. Participants were dosed with C. jejuni CG8421 and followed as inpatients. Pattern of illness, bacterial shedding, and immunologic responses were determined. Following screening, 23 subjects received 1 X 10(6) or 1 X 10(5) colony-forming units of C. jejuni, with attack rates (percentage of patients who became ill) of 100% (1 X 10(6) colony-forming units) or 93% (1 X 10(5) colony-forming units). Every subject shed CG8421; the median time to diarrhea onset was 72.3 h (interquartile range, 53.9-99.9 h). Symptoms included abdominal cramps (74%), nausea (65%), and fever (39%). No major safety concerns occurred, including bacteremia, hypotension, or postinfectious sequelae. Unexpectedly, recrudescent infection occurred in 2 subjects (1 subject without Campylobacter-specific adaptive immune responses and 1 with azithromycin resistance acquired in vivo); both infections cleared after receipt of additional antibiotics. Cumulative Campylobacter-specific immune responses were as follows: serologic response occurred in 87% (immunoglobulin [Ig] A) and 48% (IgG) of subjects, in vitro interferon-gamma production occurred in 91% of subjects, and 96% of subjects had IgA antibody-secreting cells and fecal IgA detected. The C. jejuni CG8421 challenge model provides a safe and effective tool, without the risk of Guillain-Barré syndrome. The model demonstrates high attack rates after lower doses of challenge

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

    DEFF Research Database (Denmark)

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

    1998-01-01

    ) typing of representative isolates indicated that trade of infected birds between farms was an important factor in the spread of C. coli. Phenotypic and genotypic data suggest a clonal variant of the principal outbreak type may account for the remaining cases from which C. coli was found. Conventional...... 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....

  5. Campylobacter jejuni induces acute enterocolitis in gnotobiotic IL-10-/- mice via Toll-like-receptor-2 and -4 signaling.

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    Lea-Maxie Haag

    Full Text Available BACKGROUND: Campylobacter jejuni is a leading cause of foodborne bacterial enterocolitis worldwide. Investigation of immunopathology is hampered by a lack of suitable vertebrate models. We have recently shown that gnotobiotic mice as well as conventional IL-10(-/- animals are susceptible to C. jejuni infection and develop intestinal immune responses. However, clinical symptoms of C. jejuni infection were rather subtle and did not reflect acute bloody diarrhea seen in human campylobacteriosis. METHODOLOGY/PRINCIPAL FINDINGS: In order to overcome these limitations we generated gnotobiotic IL-10(-/- mice by quintuple antibiotic treatment starting right after weaning. The early treatment was essential to prevent these animals from chronic colitis. Following oral infection C. jejuni colonized the gastrointestinal tract at high levels and induced acute enterocolitis within 7 days as indicated by bloody diarrhea and pronounced histopathological changes of the colonic mucosa. Immunopathology was further characterized by increased numbers of apoptotic cells, regulatory T-cells, T- and B-lymphocytes as well as elevated TNF-α, IFN-γ, and MCP-1 concentrations in the inflamed colon. The induction of enterocolitis was specific for C. jejuni given that control animals infected with a commensal E. coli strain did not display any signs of disease. Most strikingly, intestinal immunopathology was ameliorated in mice lacking Toll-like-receptors-2 or -4 indicating that C. jejuni lipoproteins and lipooligosaccharide are essential for induction and progression of immunopathology. CONCLUSION/SIGNIFICANCE: Gnotobiotic IL-10(-/- mice develop acute enterocolitis following C. jejuni infection mimicking severe episodes of human campylobacteriosis and are thus well suited to further dissect mechanisms underlying Campylobacter infections in vivo.

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

    Directory of Open Access Journals (Sweden)

    Hana Trigui

    2017-09-01

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

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

    International Nuclear Information System (INIS)

    Al-Amri, Ahlam I.; Bakhiet, Moiz O.; Botta, Giuseppe A.; Tabbara, Khaled S.; Ismaeel, Abdelrahman Y.; Al-Mahmeed, Ali E.; Bin Danya, Khalid M.

    2008-01-01

    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

  8. Phenotypic and Transcriptomic Responses ofCampylobacter jejuniSuspended in an Artificial Freshwater Medium.

    Science.gov (United States)

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

    2017-01-01

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

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

    Directory of Open Access Journals (Sweden)

    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

  10. Profiling of Campylobacter jejuni Proteome in Exponential and Stationary Phase of Growth

    Directory of Open Access Journals (Sweden)

    Hana Turonova

    2017-05-01

    Full Text Available Campylobacter jejuni has been reported as a major cause of bacterial food-borne enteritides in developed countries during the last decade. Despite its fastidious growth requirements, including low level of oxygen and high level of CO2, this pathogen is able to persist in the environment without permanent loss of its viability and virulence. As C. jejuni is not able to multiply outside a host, the cells spend significant amount of time in stationary phase of growth. The entry into the stationary phase is often correlated to resistance to various stresses in bacteria. The switching between exponential and stationary phases is frequently mediated by the regulator sigma S (RpoS. However, this factor is absent in C. jejuni and molecular mechanisms responsible for transition of cells to the stationary phase remain elusive. In this work, proteomic profiles of cells from exponential and stationary phases were compared using 2-D electrophoresis (2DE fingerprinting combined with mass spectrometry analysis and qRT-PCR. The identified proteins, whose expression differed between the two phases, are mostly involved in protein biosynthesis, carbon metabolism, stress response and motility. Altered expression was observed also in the pleiotropic regulator CosR that was over-expressed during stationary phase. A shift between transcript and protein level evolution of CosR throughout the growth of C. jejuni was observed using qRT-PCR and (2DE. From these data, we hypothesized that CosR could undergo a negative autoregulation in stationary phase. A consensus sequence resulting from promoter sequence alignment of genes potentially regulated by CosR, including its own upstream region, among C. jejuni strains is proposed. To verify experimentally the potential autoregulation of CosR at the DNA level, electrophoretic mobility shift assay was performed with DNA fragments of CosR promoter region and rCosR. Different migration pattern of the promoter fragments indicates

  11. Detection of small numbers of Campylobacter jejuni and Campylobacter coli cells in environmental water, sewage, and food samples by a seminested PCR assay.

    Science.gov (United States)

    Waage, A S; Vardund, T; Lund, V; Kapperud, G

    1999-04-01

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

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

    DEFF Research Database (Denmark)

    Ranjitkar, Samir; Engberg, Ricarda Greuel

    2016-01-01

    An infection trial and a production trial over 35 days were conducted in parallel to study the influence of feeding crimped kernel maize silage (CKMS) on the intestinal Campylobacter jejuni colonization and broiler performance, respectively. The CKMS was used at dietary inclusion levels of 15...... 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....

  13. Detection of non-jejuni and -coli Campylobacter species from stool specimens with an immunochromatographic antigen detection assay.

    Science.gov (United States)

    Couturier, Brianne A; Couturier, Marc Roger; Kalp, Kim J; Fisher, Mark A

    2013-06-01

    The STAT! Campy immunochromatographic assay for Campylobacter antigen was compared to culture for 500 clinical stool specimens. Antigen was detected in six culture-negative, PCR-positive specimens. C. upsaliensis, a pathogenic species that is traditionally difficult to recover in routine stool cultures, was detected in two of these culture-negative specimens. This study provides evidence that antigen testing may cross-react with at least one additional non-jejuni and -coli Campylobacter species that may be missed by routine culture for campylobacteriosis.

  14. Complete Genome Sequences of Multidrug-Resistant Campylobacter jejuni Strain 14980A (Turkey Feces) and Campylobacter coli Strain 14983A (Housefly from a Turkey Farm), Harboring a Novel Gentamicin Resistance Mobile Element

    OpenAIRE

    Miller, William G.; Huynh, Steven; Parker, Craig T.; Niedermeyer, Jeffrey A.; Kathariou, Sophia

    2016-01-01

    Multidrug resistance (MDR) in foodborne pathogens is a major food safety and public health issue. Here we describe whole-genome sequences of two MDR strains of Campylobacter jejuni and Campylobacter coli from turkey feces and a housefly from a turkey farm. Both strains harbor a novel chromosomal gentamicin resistance mobile element.

  15. Presencia de campylobacter jejuni en aves de corral y sus manipuladores

    Directory of Open Access Journals (Sweden)

    Fabio Carmona

    1985-12-01

    Full Text Available En una granja procesadora de aves en la ciudad de Cali, donde se sacrifican 12.000 animales diarios, se escogió para buscar Campylobacter jejuni una muestra de 120 pollos tomando como base una prevalencia del 80% y empleando un muestreo sistemático para dar una mayor cobertura teniendo en cuenta que la procedencia de los animales variaba diariamente. El porcentaje de positividad en materia fecal fue de 98.33% (118/120. Con la finalidad de averiguar el grado de higiene empleado en la manipulación de las canales, se frotó la superficie de ellos antes y después de introducirle las vísceras que es sabido van dentro del pollo al llegar al consumidor; se encontró que las canales con vísceras estaban contaminadas en un 18.33% (11/60 mientras que antes de este procedimiento la contaminación tan solo fue del 8.33% (5/60. También fueron examinadas 62 personas para buscar en ellas el C. jejuni encontrhndose en 5 (8.06% la bacteria. A todas las cepas aisladas se les probó la sensibilidad a la eritromicina usando disco de 15 Ug observándose un 100% de sensibilidad a la droga.

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

    Energy Technology Data Exchange (ETDEWEB)

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

    2007-01-01

    The transcriptional regulator CmeR from C. jejuni has been purified and crystallized and X-ray diffraction data have been collected to a resolution of 2.2 Å. In Campylobacter jejuni, a Gram-negative bacterial pathogen causing gastroenteritis in humans, the CmeR regulatory protein controls transcription of the multidrug transporter gene operon cmeABC. CmeR belongs to the TetR family of transcriptional regulators. The 210-residue CmeR consists of two functional motifs: an N-terminal DNA-binding domain and a C-terminal ligand-binding domain. It is predicted that the DNA-binding domain interacts directly with target promoters, while the C-terminal motif interacts with inducing ligands (such as bile salts). As an initial step towards confirming this structural model, recombinant CmeR protein containing a 6×His tag at the N-terminus was crystallized. Crystals of ligand-free CmeR belonged to space group P2{sub 1}2{sub 1}2, with unit-cell parameters a = 37.4, b = 57.6, c = 93.3 Å. Diffraction was observed to at least 2.2 Å at 100 K. Analysis of the detailed CmeR structure is currently in progress.

  17. [Suitability of SSCP-PCR analysis for molecular detection of quinolone resistance in Campylobacter jejuni].

    Science.gov (United States)

    Beckmann, Lutz; Müller, Monika; Luber, Petra; Schrader, Christina; Bartelt, Edda; Klein, Günter

    2003-01-01

    Foodborne infections with Campylobacter spp. are increasing, especially antibiotic resistant strains are emerging. Quinolone resistant isolates can cause failure of therapy in severe clinical infections. Molecular characterisation is needed for the detection of resistant variants of C. jejuni. Therefore 23 isolates from poultry and human medicine as well as three control strains were tested for their minimal inhibitory concentration, their Single-Strand-Conformation-Polymorphism (SSCP)-PCR pattern (a method for the detection of resistance determining point mutations), and their sequence of the quinolone resistance determining region (QRDR). Six different SSCP types could be identified: two types for quinolone resistant isolates and other types containing so called silent mutations without influence on the resistance. A genotypic monitoring of the quinolone resistance in C. jejuni can be useful for the early detection of new resistance variants. As a screening method for detection of point mutations in the QRDR the SSCP-PCR can be applied. Compared to other genotypic methods the SSCP-PCR is less time and cost consuming and needs only standard technical equipment.

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

    Energy Technology Data Exchange (ETDEWEB)

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

    2009-04-22

    PEB3 is a glycoprotein adhesin from Campylobacter jejuni whose structure suggested a role in transport. We have investigated potential ligands for PEB3 and characterized their binding properties using biophysical methods in solution and by X-ray crystallography. A thermal aggregation assay of PEB3 with a library of physiological compounds identified three possible ligands [3-phosphoglycerate (3-PG), phosphoenolpyruvate (PEP), and aconitate], which stabilized wild-type PEB3 but did not stabilize either a PEB3 form containing two mutations at the ligand-binding site, T138A/S139A, or a second PEB3 mutant, K135E, at a site {approx}14 {angstrom} away. Fluorescence titration experiments and cocrystal structures with various ligands were used to characterize the binding of 3-PG, PEP, and phosphate to PEB3. Further, a C. jejuni growth experiment in minimal medium supplemented with 3-PG showed that this molecule enhances the growth of wild-type C. jejuni, but not of the PEB3 mutants. Crystallographic analysis of PEB3 complexes revealed that the Ser171-Gln180 region in the presence of 3-PG or other phosphates is helical and similar to those of other transport proteins, but it is nonhelical when citrate is bound. The K135E mutation resulted in expression of a more highly glycosylated form of PEB3 in vivo, and its crystal structure showed the conformation of the first two residues of the glycan. On the basis of our findings, we suggest that PEB3 is a transport protein that may function in utilization of 3-PG or other phosphate-containing molecules from the host.

  19. Rapid Detection of Campylobacter coli, C. jejuni, and Salmonella enterica on Poultry Carcasses by Using PCR-Enzyme-Linked Immunosorbent Assay

    OpenAIRE

    Hong, Yang; Berrang, Mark E.; Liu, Tongrui; Hofacre, Charles L.; Sanchez, Susan; Wang, Lihua; Maurer, John J.

    2003-01-01

    Contamination of retail poultry by Campylobacter spp. and Salmonella enterica is a significant source of human diarrheal disease. Isolation and identification of these microorganisms require a series of biochemical and serological tests. In this study, Campylobacter ceuE and Salmonella invA genes were used to design probes in PCR-enzyme-linked immunosorbent assay (ELISA), as an alternative to conventional bacteriological methodology, for the rapid detection of Campylobacter jejuni, Campylobac...

  20. A PCR-RFLP assay for the detection and differentiation of Campylobacter jejuni, C. coli, C. fetus, C. hyointestinalis, C. lari, C. helveticus and C. upsaliensis.

    Science.gov (United States)

    Kamei, Kazumasa; Asakura, Masahiro; Somroop, Srinuan; Hatanaka, Noritoshi; Hinenoya, Atsushi; Nagita, Akira; Misawa, Naoaki; Matsuda, Motoo; Nakagawa, Shinsaku; Yamasaki, Shinji

    2014-05-01

    Although Campylobacter jejuni and Campylobacter coli are the most common bacterial causes of human gastrointestinal diseases, other Campylobacter species are also involved in human and animal infections. In this study, we developed a cytolethal distending toxin (cdt) gene-based PCR-RFLP assay for the detection and differentiation of C. jejuni, C. coli, C. fetus, C. hyointestinalis, C. lari, C. helveticus and C. upsaliensis. Previously designed common primers, which can amplify the cdtB gene of C. jejuni, C. coli and C. fetus, were used for detecting seven Campylobacter species and differentiating between them by restriction digestion. The PCR-RFLP assay was validated with 277 strains, including 35 C. jejuni, 19 C. coli, 20 C. fetus, 24 C. hyointestinalis, 13 C. lari, 2 C. helveticus, 22 C. upsaliensis, 3 other Campylobacter spp. and 17 other species associated with human diseases. Sensitivity and specificity of the PCR-RFLP assay were 100 % except for C. hyointestinalis (88 % sensitivity). Furthermore, the PCR-RFLP assay successfully detected and differentiated C. jejuni, C. coli and C. fetus in clinical and animal samples. The results indicate that the PCR-RFLP assay is useful for the detection and differentiation of seven Campylobacter species important for human and animal diseases.

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

    Directory of Open Access Journals (Sweden)

    Lavinia Ştef

    2015-05-01

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

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

    Science.gov (United States)

    Lerasle, M.; Guillou, S.; Simonin, H.; Laroche, M.; de Lamballerie, M.; Federighi, M.

    2012-03-01

    Our study focuses on a foodborne pathogen, Campylobacter, which is responsible for the most frequent bacterial enteritis worldwide. Membrane integrity of Campylobacter jejuni NCTC 11168 cells treated at high pressure (300 MPa, 20°C, 10 min) at pH 7.0 and pH 5.6 was measured by fluorescence spectroscopy of propidium iodide (PI) uptake. The percentage of membrane-damaged cells by high pressure, in which PI is allowed to penetrate, was determined using two calibration methods based on the PI fluorescence signal obtained with cells killed either by a heat treatment (80°C for 15 min) or by a pressure treatment (400 MPa, 20°C, 10 min). Both calibrations were shown to be statistically different (P<0.05), particularly at acidic pH, suggesting that a difference in the penetration of PI into bacterial cells might depend on the mode of cell inactivation. These results corroborate the fact that the mechanism of microbial inactivation by high pressure is pH-dependent.

  3. Acute disseminated encephalomyelitis following Campylobacter jejuni gastroenteritis: Case report and review of the literature.

    Science.gov (United States)

    Marziali, Simone; Picchi, Eliseo; Di Giuliano, Francesca; Altobelli, Simone; Mataluni, Giorgia; Marfia, Girolama; Garaci, Francesco; Floris, Roberto

    2017-02-01

    We describe a case of a 25-year-old male with a diagnosis of acute disseminated encephalomyelitis (ADEM) following infection with Campylobacter jejuni, which is implicated in various human pathologies regarding the central nervous system (CNS) with acute course like Guillain-Barré syndrome (GBS), Miller-Fisher syndrome (MFS), Bickerstaff's brainstem encephalitis (BEE), acute transverse myelitis (ATM) as well as ADEM. These conditions are caused by cross-reactivity between Campylobacter's epitopes and cells of the CNS that causes an immunomediated inflammatory demyelination of the CNS. In the acute phase, magnetic resonance (MR) can detect pathologic signal intensity at the CNS with areas of pathologic contrast enhancement at cortical and spinal white matter that normalize over time or can be stable. These findings can be associated with edema in parts of the CNS. The lesions typically appear at different times during the disease course and also can have a different evolution. Our purpose therefore was to describe the clinical course and MR findings of this case and perform a critical review of the literature.

  4. Potential of coriander (Coriandrum sativum) oil as a natural antimicrobial compound in controlling Campylobacter jejuni in raw meat.

    Science.gov (United States)

    Rattanachaikunsopon, Pongsak; Phumkhachorn, Parichat

    2010-01-01

    Twelve essential oils were tested in vitro for antimicrobial activities against several strains of Campylobacter jejuni, a pathogen causing food-borne diseases worldwide. Using disk diffusion and minimal inhibitory concentration determination assays, we noted that coriander oil exhibited the strongest antimicrobial activity against all tested strains. The oil had a bactericidal effect on the target bacteria. In evaluating the antimicrobial potency of coriander oil against C. jejuni on beef and chicken meat at 4 degrees C and 32 degrees C, it was found that the oil reduced the bacterial cell load in a dose-dependent manner. The type of meat and temperature did not influence the antimicrobial activity of the oil. This study indicates the potential of coriander oil to serve as a natural antimicrobial compound against C. jejuni in food.

  5. Genetic diversity and host associations in Campylobacter jejuni from human cases and broilers in 2000 and 2008.

    Science.gov (United States)

    Griekspoor, Petra; Engvall, Eva Olsson; Åkerlind, Britt; Olsen, Björn; Waldenström, Jonas

    2015-07-09

    Campylobacter jejuni is an important food-borne pathogen, with a global distribution. It can colonize numerous host species, including both domestic and wild animals, but is particularly associated with birds (poultry and wild birds). For human campylobacteriosis, poultry products are deemed the most significant risk factor for acquiring infection. We conducted a genotyping and host attribution study of a large representative collection of C. jejuni isolated from humans and broilers in Sweden in the years 2000 and 2008. In total 673 broiler and human isolates from 10 different abattoirs and 6 different hospitals were genotyped with multilocus sequence typing. Source attribution analyses confirmed the strong linkage between broiler C. jejuni and domestic human cases, but also indicated a significant association to genotypes more commonly found in wild birds. Genotype distributions did not change dramatically between the two study years, suggesting a stable population of infecting bacteria. Copyright © 2015 Elsevier B.V. All rights reserved.

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

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    Euna eOh

    2015-04-01

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

  7. Real-time PCR assay for rapid detection and quantification of Campylobacter jejuni on chicken rinses from poultry processing plant.

    Science.gov (United States)

    Debretsion, Aradom; Habtemariam, Tsegaye; Wilson, Saul; Nganwa, David; Yehualaeshet, Teshome

    2007-06-01

    Campylobacter jejuni (C. jejuni) is the leading cause of food-borne gastroenteritis in the United States. Detection of Campylobacter in food samples by conventional culture is cumbersome; therefore, there is a need to develop rapid and cost-effective detection and quantification methods. Eighty-four whole chicken rinses were collected at different stages of processing at three poultry processing plants. After chicken wash collection and DNA extraction, the samples were directly subjected to real-time PCR (rtPCR) without enrichment and also culture. The assay specificity was determined with a range of Campylobacter species, related, and unrelated organisms. Of the 84 samples collected 65 (77%) of the samples were positive by the rtPCR assay and 27 (32%) of the samples tested positive by direct plating to selective agar media. The results were positively concordant for 27 (32%) of the samples. The whole rtPCR assay can be completed within 90min with a detection limit of 1CFU, compared to 5-7 days for enrichment and sub culturing in selective agar. This assay is the first report of rtPCR method capable of detecting and quantifying C. jejuni from chicken rinses without an enrichment step and could be an important, rapid and quantification model for other food-borne pathogens.

  8. Effects of efflux-pump inducers and genetic variation of the multidrug transporter cmeB in biocide resistance of Campylobacter jejuni and Campylobacter coli.

    Science.gov (United States)

    Mavri, Ana; Smole Možina, Sonja

    2013-03-01

    Multidrug efflux pumps, such as CmeABC and CmeDEF, are involved in the resistance of Campylobacter to a broad spectrum of antimicrobials. The aim of this study was to analyse the effects of two putative efflux-pump inducers, bile salts and sodium deoxycholate, on the resistance of Campylobacter to biocides (triclosan, benzalkonium chloride, chlorhexidine diacetate, cetylpyridinium chloride and trisodium phosphate), SDS and erythromycin. The involvement of the CmeABC and CmeDEF efflux pumps in this resistance was studied on the basis of the effects of bile salts and sodium deoxycholate in Campylobacter cmeB, cmeF and cmeR mutants. The genetic variation in the cmeB gene was also examined, to see whether this polymorphism is related to the function of the efflux pump. In 15 Campylobacter jejuni and 23 Campylobacter coli strains, bile salts and sodium deoxycholate increased the MICs of benzalkonium chloride, chlorhexidine diacetate, cetylpyridinium chloride and SDS, and decreased the MICs of triclosan, trisodium phosphate and erythromycin. Bile salts and sodium deoxycholate further decreased or increased the MICs of biocides and erythromycin in the cmeF and cmeR mutants. For cmeB polymorphisms, 17 different cmeB-specific PCR-RFLP patterns were identified: six within C. jejuni only, nine within C. coli only and two in both species. In conclusion, bile salts and sodium deoxycholate can increase or decrease bacterial resistance to structurally unrelated antimicrobials. The MIC increases in the cmeF and cmeR mutants indicated that at least one non-CmeABC efflux system is involved in resistance to biocides. These results indicate that the cmeB gene polymorphism identified is not associated with biocide and erythromycin resistance in Campylobacter.

  9. Temporal induction of pro-inflammatory and regulatory cytokines in human peripheral blood mononuclear cells by Campylobacter jejuni and Campylobacter coli.

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    Eman Hamza

    Full Text Available Campylobacter jejuni along with C. coli are major cause of human gastroenteritis worldwide. So far, the human immune response against Campylobacter is not entirely clear. We hypothesize that it is coordinated by an interaction between pro-inflammatory and regulatory cytokines which is influenced by bacterial and host-individual differences. Accordingly, we used peripheral blood mononuclear cells (PBMC from healthy donors to study the primary systemic immune response to C. jejuni and C. coli. PBMC were stimulated by different strains of C. jejuni and C. coli for three time points (5, 10, 24 hours. The production of the pro-inflammatory (IL-6, IL-8, IFN-γ and the regulatory (IL-10 cytokines were measured by ELISA. All strains induced higher levels of IL-8 and IL-6 than IFN-γ and IL-10. In contrast to IL-8 and IL-6, IL-10 showed a steeper increase over time. While IFN-γ did not show any further increase between 10 and 24 hours. Interestingly, there was a significant correlation between IL-8 and IL-10 which peaked at 24 hours. Despite the variability of the used bacterial strains, their effect on cytokine production was less pronounced than the inter-person differences. The strongest significant effect of the strain was on the level of IL-10. IL-10 and IL-6 were significantly influenced by strain-person interaction. In conclusion, the systemic immune response to C. coli and C. jejuni is characterized by an early pro-inflammatory reaction with later initiation of regulatory immune response which is influenced mainly by the host, explaining the individual variations in disease severity. Additional work is needed to determine the cellular sources of the produced cytokines as well as the campylobacter molecules that might contribute to this stimulation.

  10. Molecular subtyping of Campylobacter jejuni subsp. jejuni strains isolated from different animal species in the state of São Paulo, Brazil Subtipagem molecular de estirpes de Campylobacter jejuni subsp. jejuni isoladas de diferentes espécies animais do Estado de São Paulo, Brasil

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    Eliana Scarcelli

    2005-12-01

    Full Text Available The objective of the present trial was to characterize genetically strains of Campylobacter jejuni subsp. jejuni isolated from humans and several animal sources (bovines, swine, dogs, primates, wild boars and poultry. A total of 828 different animal samples (feces, carcass, aborted fetus and hysterectomized uterus were analysed by means of routine bacteriological methods, and 36 C. jejuni strains were isolated. Thirty strains of human fecal origin were obtained in clinical analysis laboratories in the city of São Paulo. The 66 C. jejuni strains isolated were submitted to genetic characterization. Primers based on fla A gene were used in a polymerase chain reaction (PCR and amplified a fragment of the 702 bp. PCR products were evaluated by means of sequencing and genealogic analysis. Genetic variability analysis of 66 strains showed 44 different subtypes of C. jejuni. One subtype was identical to a C. jejuni strain of human origin with the sequence in the GenBank (GENBANK accession number AF050186. Subtyping analysis of C. jejuni strains based on sequencing of the fla A gene variable region and analysis of sequence alignment by the Maximum Parsimony method showed to be highly discriminatory, providing the best conditions to differentiate strains involved in outbreaks from those sporadically isolated. This is the first study of molecular subtyping analysis of human and animal C. jejuni strains using sequencing technique and genealogic analysis in the state of São Paulo, Brazil.O objetivo do presente trabalho foi caracterizar geneticamente estirpes de Campylobacter jejuni subsp. jejuni isoladas de humanos e de diferentes origens animais (bovinas, suínas, cães, primatas, javalis, suínos e aves de corte. Um total de 828 amostras (fezes, carcaças, fetos abortados e útero histerectomizado foram analisadas por métodos de rotina bacteriológica e 36 estirpes de C. jejuni foram isoladas. Trinta estirpes de origem fecal humana foram obtidas de

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

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    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.

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

    DEFF Research Database (Denmark)

    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 all C...

  13. Phenotypic and genetic analyses of Campylobacter jejuni Lior serotype 76 isolated from chicken meat and clinical specimens.

    Science.gov (United States)

    Matsumoto, Masakado; Hiramatsu, Reiji; Yamada, Kazuhiro; Suzuki, Masahiro; Miwa, Yoshio; Yabutani, Mitsutaka; Nagai, Yuhki; Tsuchiya, Michiyo; Noda, Makiko; Nagata, Akihiro; Kawakami, Keiko; Shima, Tomoko; Tatsumi, Norio; Minagawa, Hiroko

    2013-01-01

    The aim of this study was to examine the link between Campylobacter jejuni isolates obtained from chicken meat (n = 7) and gastroenteritis patients (n = 744). In total, 751 isolates were subjected to Lior serotyping. All the isolates from chicken meats were serotyped as Lior serotype 76 (LIO76). Among 23 of the identified LIO76 strains, 13 strains (6 from chicken meat and 7 from clinical specimens) were indistinguishable by Penner serotyping, antimicrobial susceptibility testing, and pulsed-field gel electrophoresis. These strains were isolated in 2 different Japanese prefectures in 2004-2005, suggesting that chicken meat is an etiological agent of Campylobacter gastroenteritis and that a diffuse outbreak occurred during this time. Therefore, a continuous surveillance program should be established in Japan in order to prevent Campylobacter gastroenteritis, especially large-scale food-borne outbreaks.

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

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    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

  15. Analysis of Campylobacter jejuni infection in the gnotobiotic piglet and genome-wide identification of bacterial factors required for infection.

    Science.gov (United States)

    de Vries, Stefan P W; Linn, Aileen; Macleod, Kareen; MacCallum, Amanda; Hardy, Simon P; Douce, Gill; Watson, Eleanor; Dagleish, Mark P; Thompson, Hal; Stevenson, Andy; Kennedy, David; Baig, Abiyad; Coward, Chris; Maskell, Duncan J; Smith, David G E; Grant, Andrew J; Everest, Paul

    2017-03-10

    To investigate how Campylobacter jejuni causes the clinical symptoms of diarrhoeal disease in humans, use of a relevant animal model is essential. Such a model should mimic the human disease closely in terms of host physiology, incubation period before onset of disease, clinical signs and a comparable outcome of disease. In this study, we used a gnotobiotic piglet model to study determinants of pathogenicity of C. jejuni. In this model, C. jejuni successfully established infection and piglets developed an increased temperature with watery diarrhoea, which was caused by a leaky epithelium and reduced bile re-absorption in the intestines. Further, we assessed the C. jejuni genes required for infection of the porcine gastrointestinal tract utilising a transposon (Tn) mutant library screen. A total of 123 genes of which Tn mutants showed attenuated piglet infection were identified. Our screen highlighted a crucial role for motility and chemotaxis, as well as central metabolism. In addition, Tn mutants of 14 genes displayed enhanced piglet infection. This study gives a unique insight into the mechanisms of C. jejuni disease in terms of host physiology and contributing bacterial factors.

  16. Standing genetic variation in contingency loci drives the rapid adaptation of Campylobacter jejuni to a novel host.

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    John P Jerome

    2011-01-01

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

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

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    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.

  18. Double-locus sequence typing using porA and peb1A for epidemiological studies of Campylobacter jejuni.

    Science.gov (United States)

    Ahmed, Monir U; Dunn, Louise; Valcanis, Mary; Hogg, Geoff; Ivanova, Elena P

    2014-03-01

    Campylobacter jejuni is the leading cause of foodborne bacterial gastroenteritis worldwide. Bacterial typing schemes play an important role in epidemiological investigations to trace the source and route of transmission of the infectious agent by identifying outbreak and differentiating among sporadic infections. In this study, a double-locus sequence typing (DLST) scheme for C. jejuni based on concatenated partial sequences of porA and peb1A genes is proposed. The DLST scheme was validated using 50 clinical and environmental C. jejuni strains isolated from human (C5, H, H15-H19), chicken (CH1-CH15), water (W2-W17), and ovine samples (OV1-OV6). The scheme was found to be highly discriminatory (discrimination index [DI]=0.964) and epidemiologically concordant based on C. jejuni strains studied. The DLST showed discriminatory power above 0.95 and excellent congruence to multilocus sequence typing and can be recommended as a rapid and low-cost typing scheme for epidemiological investigation of C. jejuni. It is suggested that the DLST scheme is suitable for identification of outbreak strains and differentiation of the sporadic infection strains.

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

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    Kelly A Fimlaid

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

  20. Comparative Proteomics Reveals Differences in Host-Pathogen Interaction between Infectious and Commensal Relationship with Campylobacter jejuni

    Directory of Open Access Journals (Sweden)

    Juan J. Garrido

    2017-04-01

    Full Text Available Campylobacter jejuni is the leading food-borne poisoning in industrialized countries. While the bacteria causes disease in humans, it merely colonizes the gut in poultry or pigs, where seems to establish a commensal relationship. Until now, few studies have been conducted to elucidate the relationship between C. jejuni and its different hosts. In this work, a comparative proteomics approach was used to identify the underlying mechanisms involved in the divergent outcome following C. jejuni infection in human and porcine host. Human (INT-407 and porcine (IPEC-1 intestinal cell lines were infected by C. jejuni for 3 h (T3h and 24 h (T24h. C. jejuni infection prompted an intense inflammatory response at T3h in human intestinal cells, mainly characterized by expression of proteins involved in cell spreading, cell migration and promotion of reactive oxygen species (ROS. Proteomic analysis evidenced significantly regulated biofunctions in human cells related with engulfment and endocytosis, and supported by canonical pathways associated to infection such as caveolar- and clathrin-mediated endocytosis signaling. In porcine IPEC-1 cells, inflammatory response as well as signaling pathways that control cellular functions such as cell migration, endocytosis and cell cycle progression resulted downregulated. These differences in the host response to infection were supported by the different pattern of adhesion and invasion proteins expressed by C. jejuni in human and porcine cells. No marked differences in expression of virulence factors involved in adaptive response and iron acquisition functions were observed. Therefore, the results of this study suggest that both host and pathogen factors are responsible for commensal or infectious character of C. jejuni in different hosts.

  1. High Prevalence of Hyper-Aerotolerant Campylobacter jejuni in Retail Poultry with Potential Implication in Human Infection.

    Science.gov (United States)

    Oh, Euna; McMullen, Lynn; Jeon, Byeonghwa

    2015-01-01

    Campylobacter jejuni is a leading cause of foodborne illnesses around the world. Since C. jejuni is microaerophilic and sensitive to oxygen, aerotolerance is important in the transmission of C. jejuni to humans via foods under aerobic conditions. In this study, 70 C. jejuni strains were isolated from retail raw chicken meats and were subject to multilocus sequence typing (MLST) analysis. In the aerotolerance testing by aerobic shaking at 200 rpm, 50 (71.4%) isolates survived after 12 h (i.e., aerotolerant), whereas 20 (28.6%) isolates did not (i.e., aerosensitive). Interestingly, further aerobic cultivation showed that 25 (35.7%) isolates still survived even after 24 h of vigorous aerobic shaking (i.e., hyper-aerotolerant). Compared to aerosensitive strains, the hyper-aerotolerant strains exhibited increased resistance to oxidative stress, both peroxide and superoxide. A mutation of ahpC in hyper-aerotolerant strains significantly impaired aerotolerance, indicating oxidative stress defense plays an important role in hyper-aerotolerance. The aerotolerant and hyper-aerotolerant strains were primarily classified into MLST clonal complexes (CCs)-21 and -45, which are known to be the major CCs implicated in human gastroenteritis. Compared to the aerosensitive strains, CC-21 was more dominant than CC-45 in aerotolerant and hyper-aerotolerant strains. The findings in this study revealed that hyper-aerotolerant C. jejuni is highly prevalent in raw chicken meats. The enhanced aerotolerance in C. jejuni would impact human infection by increasing possibilities of the foodborne transmission of C. jejuni under aerobic conditions.

  2. Evaluation and single-laboratory verification of a proposed modification to the U.S. Food and Drug Administration method for detection and identification of Campylobacter jejuni or Campylobacter coli from raw silo milk.

    Science.gov (United States)

    Gharst, Greg; Bark, Don H; Newkirk, Robert; Guillen, Lacey; Wang, Qian; Abeyta, Carlos

    2013-01-01

    The current U.S. Food and Drug Administration (FDA) methodology for detection of Campylobacter, a leading source for foodborne illness, is outdated. The purpose of this study, therefore, was to improve and update the cultural and identification methods found in the FDA/Bacteriological Analytical Manual (BAM). Raw silo milk samples containing typical and atypical strains of Campylobacter jejuni and Campylobacter coli at different levels (5 CFU/25 g, 50 CFU/25 g, and 125 CFU/25 g) were analyzed. Valid results were obtained from 240 test portions. Six inoculated (at the levels described above) and two uninoculated samples were sent to a participating laboratory to mimic a "real-world" scenario. These combined data indicated that the use of sheep blood in combination with enrichment is not necessary. R & F Campylobacter jejuni/Campylobacter coli Chromogenic Plating Medium is significantly (P detection of C. jejuni or C. coli at low inoculation levels than the modified Cefoperazone Charcoal Deoxycholate Agar used in the BAM. The quantitative PCR method described demonstrated rapid confirmation and identification of C. jejuni or C. coli. It reduced the time to isolate C. jejuni or C. coli, and increased the sensitivity compared to the current BAM protocol.

  3. Development and evaluation of internal amplification controls for use in a real-time duplex PCR assay for detection of Campylobacter coli and Campylobacter jejuni.

    Science.gov (United States)

    Randall, Luke; Lemma, Fabrizio; Rodgers, John; Vidal, Ana; Clifton-Hadley, Felicity

    2010-02-01

    A common problem of both conventional and real-time PCR assays is failure of DNA amplification due to the presence of inhibitory substances in samples. In view of this, our aim was to develop and evaluate internal amplification controls (IACs) for use with an existing duplex real-time PCR assay for Campylobacter coli and Campylobacter jejuni. Both competitive and non-competitive IACs were developed and evaluated. The competitive approach involved a DNA fragment of the coding region of the fish viral haemorrhagic septicaemia virus, flanked by the mapA PCR primers, whilst the non-competitive approach utilized an extra set of universal 16S rDNA primers. Both IAC-PCR assay types were evaluated using cultures of Campylobacter and chicken caecal content samples. Both IACs were sensitive to caecal inhibitors, making them suitable for detecting inhibition which could lead to false-negatives. Results showed that both IACs at optimum concentrations worked well without reducing the overall sensitivity of the PCR assay. Compared to culture, the optimized competitive IAC-PCR assay detected 45/47 positives (sensitivity 93.6 %, specificity 80.1 %); however, it had the advantage over culture in that it could detect mixed infections of C. coli and C. jejuni and was capable of giving a result for a sample within a day.

  4. Antimicrobial Resistance and Multilocus Sequence Types of Finnish Campylobacter jejuni Isolates from Multiple Sources.

    Science.gov (United States)

    Olkkola, S; Nykäsenoja, S; Raulo, S; Llarena, A-K; Kovanen, S; Kivistö, R; Myllyniemi, A-L; Hänninen, M-L

    2016-02-01

    Antimicrobial susceptibility was determined for 805 domestic Campylobacter jejuni isolates obtained from broilers (n = 459), bovines (n = 120), human patients (n = 95), natural waters (n = 80), wild birds (n = 35) and zoo animals/enclosures (n = 16) with known multilocus sequence types (MLST) for 450 isolates. The minimum inhibitory concentration (MIC) values for erythromycin, tetracycline, streptomycin, gentamicin and the quinolones ciprofloxacin and nalidixic acid were determined with the VetMIC method. MICs were compared with MLST types to find possible associations between sequence type and resistance. The proportions of resistant isolates were 5% (broilers), 6.3% (natural waters), 11.4% (wild birds), 11.6% (human patients), 16.7% (bovines) and 31.3% (zoo). The most common resistance among the human and bovine isolates was quinolone resistance alone while resistance to streptomycin alone was most often detected among the broiler isolates and tetracycline resistance was most commonly observed in the wild bird, water and zoo isolates. No or negligible resistance to erythromycin or gentamicin was detected. In all data, 12/26 of the tetracycline-resistant isolates were also resistant to streptomycin (P resistant isolates, most originating from the zoo and broilers with closely associated MLST types from these sources. No association between quinolone resistance and MLST type was seen. The low percentage of resistant isolates among the domestic Campylobacter infections is most probably due to the long-term controlled use of antimicrobials. However, the higher percentage of tetracycline resistance observed among the zoo isolates could present a risk for zoo visitors of acquisition of resistant C. jejuni. The resistance pattern of tetracycline and streptomycin most often found in ST-1034 CC could indicate a common resistance acquisition mechanism commonly present in this CC. Overall, MLST typing was found to be a useful method in recognition of potential

  5. Molecular detection identified a type six secretion system in Campylobacter jejuni from various sources but not from human cases.

    Science.gov (United States)

    Siddiqui, F; Champion, O; Akram, M; Studholme, D; Eqani, S A M A S; Wren, B W; Titball, R; Bokhari, H

    2015-05-01

    To determine the presence of the T6SS in Campylobacter jejuni from diverse sources. The recently identified type VI secretion system (T6SS) is a bacterial injection machinery that plays a role in virulence, symbiosis, bacterial interactions and environmental stress responses. This system has been recently discovered in the major enteric pathogen Camp. jejuni. In this study, we used multiplex PCR (mPCR), based on conserved genetic markers of the T6SS, to screen 366 Pakistani Camp. jejuni isolates from humans, poultry, cattle, wildlife or waste-water sources. We identified the T6SS in isolates from all of these sources except humans. The overall prevalence of the T6SS among the isolates was 17/366 (4·6%) and the T6SS positive isolates clustered into four different groups. Transcription of the T6SS genes, determined using RT-PCR, was observed in bacteria cultured at 37 or 42°C but not in 37°C cultures adjusted to pH3. Campylobacter jejuni isolates harbouring T6SS markers genes were identified in livestock and non-livestock sources but in this study we did not identify human diarrhoeal isolates which possessed the T6SS. We demonstrated down-regulation of T6SS in an acidic environment. This study questions the role of the T6SS in human diarrhoeal disease. Moreover this study did not identify a clear association of Camp. jejuni isolates harbouring T6SS with any of the niches tested. Our study highlights the need to establish the role of the T6SS in environmental survival or virulence. © 2015 The Society for Applied Microbiology.

  6. Development of a modified gentamicin protection assay to investigate the interaction between Campylobacter jejuni and Acanthamoeba castellanii ATCC 30010.

    Science.gov (United States)

    Dirks, Brian P; Quinlan, Jennifer J

    2014-05-01

    Campylobacter jejuni is one of the leading causes of diarrheal illness worldwide. It is persistent in the environment and on poultry despite its microaerophilic nature and sensitivity to dessication and pH. Studies have demonstrated that C. jejuni co-incubated with Acanthamoeba spp. may be protected from harmful environmental factors. Research in this area, however has included a range of different methodologies for co-incubation, recovery of bacteria and amoebae, and verification of internalization. In this study a modified gentamicin protection assay (mGPA) was developed with a standardized co-incubation procedure. The mGPA addresses limitations of the traditional GPA by providing quantification of the rate of internalization, or lack of internalization, of C. jejuni by Acanthamoeba castellanii. The mGPA described here utilizes tubes instead of cell culture plates allowing for determination of exact numbers of A. castellanii and C. jejuni to be co-incubated prior to addition to tubes. Additionally, the mGPA allows for the incorporation of C. jejuni-only controls to determine the fate of C. jejuni throughout the assay in the absence of A. castellanii. Using the mGPA it was determined that on average 1.6×10(5) C. jejuni (or 0.006% of initial 1×10(9) inoculum) survive the assay in the absence of A. castellanii. Additionally, results obtained with the mGPA demonstrated that while co-incubation with amoebae sometimes (56% of co-incubations) provided a protective effect for C. jejuni, in other cases it did not provide any protective effect (39% of co-incubations), and in at least one case there was a statistically significant higher recovery of C. jejuni in controls when compared to C. jejuni co-incubated with amoebae. The modified gentamicin protection assay described here allows better quantification of the rate and incidence of internalization of bacteria by amoebae. Use of the standardized mGPA developed here with varying environmental parameters and/or strains

  7. Development of a loop-mediated isothermal amplification assay for rapid, sensitive detection of Campylobacter jejuni in cattle farm samples.

    Science.gov (United States)

    Dong, Hee-Jin; Cho, Ae-Ri; Hahn, Tae-Wook; Cho, Seongbeom

    2014-09-01

    Campylobacter jejuni is a leading cause of bacterial foodborne disease worldwide. The detection of this organism in cattle and their environment is important for the control of C. jejuni transmission and the prevention of campylobacteriosis. Here, we describe the development of a rapid and sensitive method for the detection of C. jejuni in naturally contaminated cattle farm samples, based on real-time loop-mediated isothermal amplification (LAMP) of the hipO gene. The LAMP assay was specific (100% inclusivity and exclusivity for 84 C. jejuni and 41 non-C. jejuni strains, respectively), sensitive (detection limit of 100 fg/μl), and quantifiable (R(2) = 0.9133). The sensitivity of the LAMP assay was then evaluated for its application to the naturally contaminated cattle farm samples. C. jejuni strains were isolated from 51 (20.7%) of 246 cattle farm samples, and the presence of the hipO gene was tested using the LAMP assay. Amplification of the hipO gene by LAMP within 30 min (mean ~10.8 min) in all C. jejuni isolates (n = 51) demonstrated its rapidity and accuracy. Next, template DNA was prepared from a total of 186 enrichment broth cultures of cattle farm samples either by boiling or using a commercial kit, and the sensitivity of detection of C. jejuni was compared between the LAMP and PCR assays. In DNA samples prepared by boiling, the higher sensitivity of the LAMP assay (84.4%) compared with the PCR assay (35.5%) indicates that it is less susceptible to the existence of inhibitors in sample material. In DNA samples prepared using a commercial kit, both the LAMP and PCR assays showed 100% sensitivity. We anticipate that the use of this rapid, sensitive, and simple LAMP assay, which is the first of its kind for the identification and screening of C. jejuni in cattle farm samples, may play an important role in the prevention of C. jejuni contamination in the food chain, thereby reducing the risk of human campylobacteriosis.

  8. The Xer/dif site-specific recombination system of Campylobacter jejuni.

    Science.gov (United States)

    Leroux, Maxime; Rezoug, Zoulikha; Szatmari, George

    2013-10-01

    Chromosome dimers, which form during the bacterial life cycle, represent a problem that must be solved by the bacterial cell machinery so that chromosome segregation can occur effectively. The Xer/dif site-specific recombination system, utilized by most bacteria, resolves chromosome dimers into monomers using two tyrosine recombinases, XerC and XerD, to perform the recombination reaction at the dif site which consists of 28-30 bp. However, single Xer recombinase systems have been recently discovered in several bacterial species. In Streptococci and Lactococci a single recombinase, XerS, is capable of completing the monomerisation reaction by acting at an atypical dif site called dif SL (31 bp). It was recently shown that a subgroup of ε-proteobacteria including Campylobacter spp. and Helicobacter spp. had a phylogenetically distinct Xer/dif recombination system with only one recombinase (XerH) and an atypical dif motif (difH). In order to biochemically characterize this system in greater detail, Campylobacter jejuni XerH was purified and its DNA-binding activity was characterized. The protein showed specific binding to the complete difH site and to both halves separately. It was also shown to form covalent complexes with difH suicide substrates. In addition, XerH was able to catalyse recombination between two difH sites located on a plasmid in Escherichia coli in vivo. This indicates that this XerH protein performs a similar function as the related XerS protein, but shows significantly different binding characteristics.

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

    DEFF Research Database (Denmark)

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

    2005-01-01

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

  10. Resistance of Campylobacter jejuni Isolated from Layer Farms in Northern Jordan Using Microbroth Dilution and Disc Diffusion Techniques.

    Science.gov (United States)

    Al-Natour, Mohammad Q; Alaboudi, Akram R; Osaili, Tareq M; Obaidat, Mohammad M

    2016-07-01

    Campylobacter jejuni is an important pathogen of significant public health importance. This pathogen is associated with human infection and has been isolated from mammals and birds. Ninety-two cloacal C. jejuni isolates were identified from 35 layer farms in Northern Jordan. Antimicrobial susceptibility was determined using minimal inhibitory concentration (MIC) and disc diffusion techniques with variable suggested breakpoints. Using MIC and EUCAST cut-off values, the study revealed a significantly high resistance level (100%) among the layers' isolates against ciprofloxacin and tetracycline. A relatively high resistance (41%) toward gentamicin and amoxicillin and low resistance to nalidixic acid (21%), erythromycin (14%), and florfenicol (6.5%) were also found. This high level of resistance may indicate abuses in the handling of antibiotics, which may require stricter control in the local layer industry. A good agreement between the 2 techniques used was demonstrated and the disc diffusion technique could be used as a rapid screening test for antimicrobial susceptibility of C. jejuni to many antibiotics using specific Campylobacter breakpoints. © 2016 Institute of Food Technologists®

  11. Problems in isolation of Campylobacter jejuni from frozen-stored raw milk and bovine fecal samples: genetic confirmation using multiplex PCR.

    Science.gov (United States)

    Murinda, Shelton E; Nguyen, Lien T; Oliver, Stephen P

    2004-01-01

    The objectives of this study were to evaluate the use of various protocols for the isolation of Campylobacter jejuni from bulk tank milk and bovine fecal samples that were stored frozen for varying times, and to develop a rapid DNA-based protocol that distinguishes C. jejuni from other thermophilic Campylobacter spp. The pathogen was recovered from fecal samples that had been stored for 96-251 days at -20 degrees C with glycerol as the cryopreservative. In a separate study, C. jejuni-positive bovine fecal samples were stored at 5 degrees C for up to 70 days without compromising subsequent recovery of the pathogen. However, the pathogen could not be recovered from pathogenpositive fecal samples stored with or without glycerol (5 mL/11 g sample) for 21 days at -20 degrees C. Bolton broth (BB) and Bolton broth with 5% blood (BBB), containing BB supplement, were used for enrichment. Bacterial isolation was achieved by streaking from BB and BBB, and filtering from BBB onto blood-free charcoal cefoperazone deoxycholate agar (CCDA). The use of BB for the recovery of Campylobacter was more sensitive than BBB, and streaking achieved better isolation rates than filtration. Multiplex PCR incorporating thermophilic Campylobacter-specific 23S rRNA and C. jejuni-specific hippuricase gene sequences was used to confirm C. jejuni. All 265 bulk tank milk samples analyzed were negative for C. jejuni, whereas five of 411 (1.2%) fecal samples tested positive. This is the first report that has used a combination of sequences of the two genes in a multiplex format to identify C. jejuni to the species level. The method described has potential for routine use in the detection of thermophilic Campylobacter in farm environmental samples as well as other samples.

  12. Rapid PCR using nested primers of the 16S rRNA and the hippuricase (hipO) genes to detect Campylobacter jejuni and Campylobacter coli in environmental samples

    DEFF Research Database (Denmark)

    Bang, Dang Duong; Wedderkopp, A.; Pedersen, Karl

    2002-01-01

    to detect Campylobacter jejuni and Campylobacter coli in environmental samples. The sensitivity of the nested PCR was determined to be 0.01 pg/PCR, corresponding to 2-3 colony forming units (cfu) per ml. The nested PCR assays were applied to detect C. jejuni and C. coli in 269 environmental samples......Identification of sources Campylobacter infection in the poultry houses is in general problematic due to the lack of reliable methods to detect campylobacteria in environmental samples. Detection of campylobacteria in environmental samples by conventional culture methods is difficult and of limited...... collected from ten broiler farms. The sensitivity, specificity and the usefulness of the PCR assay for detection of C. jejuni and C coli in environmental samples are presented and discussed....

  13. Clinical Onset of Celiac Disease after an Episode of Campylobacter jejuni Enteritis

    Directory of Open Access Journals (Sweden)

    EF Verdu

    2007-01-01

    Full Text Available The present report describes a young woman with no previous gastrointestinal complaints who was initially diagnosed with postinfective irritable bowel syndrome (IBS after a confirmed case of Campylobacter jejuni enteritis. However, because of persistent diarrhea, new-onset bloating and the development of iron and vitamin deficiencies, serological markers for celiac disease (CD were evaluated. A positive tissue transglutaminase immunoglobulin A antibody test and repeat endoscopy with duodenal biopsy showing a Marsh IIIa lesion confirmed the diagnosis of CD. Infectious gastroenteritis is a well-established risk factor for the development of IBS, and there is recent evidence that it could play a role in the initiation and exacerbation of inflammatory bowel disease. The present case suggests that the clinical expression of CD can be unmasked by an acute gastrointestinal infection and supports the hypothesis that environmental factors other than gliadin may play a role in the clinical onset of CD in a genetically susceptible host. The increasing availability of serological testing and upper endoscopy has led to increasingly frequent diagnoses of CD and recognition that it may mimic IBS. The present case findings suggest that CD should be considered in the differential diagnosis of persistent IBS-like symptoms after an episode of infectious gastroenteritis.

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

    Energy Technology Data Exchange (ETDEWEB)

    Olivier, N.; Imperiali, B

    2008-01-01

    The carbohydrate 2, 4-diacetamido-2, 4, 6-trideoxy-{alpha}-d-glucopyranose (BacAc2) is found in a variety of eubacterial pathogens. In Campylobacter jejuni, PglD acetylates the C4 amino group on UDP-2-acetamido-4-amino-2, 4, 6-trideoxy-a-d-glucopyranose (UDP-4-amino-sugar) to form UDP-BacAc2. Sequence analysis predicts PglD to be a member of the left-handed {Beta} helix family of enzymes. However, poor sequence homology between PglD and left-handed {Beta} helix enzymes with existing structural data precludes unambiguous identification of the active site. The co-crystal structures of PglD in the presence of citrate, acetyl coenzyme A, or the UDP-4-amino-sugar were solved. The biological assembly is a trimer with one active site formed between two protomers. Residues lining the active site were identified, and results from functional assays on alanine mutants suggest His-125 is critical for catalysis, whereas His-15 and His-134 are involved in substrate binding. These results are discussed in the context of implications for proteins homologous to PglD in other pathogens.

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

    Energy Technology Data Exchange (ETDEWEB)

    Su, Chih-Chia; Yin, Linxiang; Kumar, Nitin; Dai, Lei; Radhakrishnan, Abhijith; Bolla, Jani Reddy; Lei, Hsiang-Ting; Chou, Tsung-Han; Delmar, Jared A.; Rajashankar, Kanagalaghatta R.; Zhang, Qijing; Shin, Yeon-Kyun; Yu, Edward W. (Cornell); (Iowa State)

    2017-08-01

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

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

    Directory of Open Access Journals (Sweden)

    N. CORCIONIVOSCHI

    2009-05-01

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

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

    Directory of Open Access Journals (Sweden)

    N. CORCIONIVOSCHI

    2009-05-01

    Full Text Available Recently, the complete genome sequence of Campylobacter jejuni NCTC 11168 was published 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.

  18. Use of Culture, PCR Analysis, and DNA Microarrays for Detection of Campylobacter jejuni and Campylobacter coli from Chicken Feces

    OpenAIRE

    Keramas, Georgios; Bang, Dang Duong; Lund, Marianne; Madsen, Mogens; Bunkenborg, Henrik; Telleman, Pieter; Christensen, Claus Bo Vöge

    2004-01-01

    A DNA microarray for detection of Campylobacter spp. was recently developed and applied to detect Campylobacter spp. directly from chicken feces. Sixty-five pooled chicken cloacal swab samples from 650 individual broiler chickens were included in the study. The results of Campylobacter sp. detection obtained with DNA microarrays were compared to those obtained by conventional culture and gel electrophoresis. By conventional culture, 60% of the samples were positive for either Campylobacter je...

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

    DEFF Research Database (Denmark)

    Ramonaite, Sigita; Kudirkiene, Egle; Tamuleviciene, Egle

    2014-01-01

    This study aimed to investigate the prevalence of Campylobacter jejuni in potential contamination sources that are not regularly monitored such as free-living urban pigeons and crows, dogs, cats and urban environmental water and to assess the possible impact on the epidemiology of campylobacterio......This study aimed to investigate the prevalence of Campylobacter jejuni in potential contamination sources that are not regularly monitored such as free-living urban pigeons and crows, dogs, cats and urban environmental water and to assess the possible impact on the epidemiology...

  20. Microbial ecology of Campylobacter jejuni in a United Kingdom chicken supply chain: intermittent common source, vertical transmission, and amplification by flock propagation.

    OpenAIRE

    Pearson, A D; Greenwood, M H; Feltham, R K; Healing, T D; Donaldson, J; Jones, D M; Colwell, R R

    1996-01-01

    A study of Campylobacter jejuni on a broiler chicken farm between 1989 and 1994 gave an estimated isolation rate of 27% (3,304 of 12,233) from a 0.9% sample of 1.44 million broiler chickens from six to eight sheds over 32 consecutive rearing flocks comprising 251 broiler shed flocks. During the study, C. jejuni was found in 35.5% of the 251 shed flocks but only 9.2% (23 of 251) had Campylobacter isolates in successive flocks, with 9 of those 23 sheds having the same serotype between consecuti...

  1. Importance of Campylobacter jejuni FliS and FliW in Flagella Biogenesis and Flagellin Secretion

    Directory of Open Access Journals (Sweden)

    Katarzyna A. Radomska

    2017-06-01

    Full Text Available Flagella-driven motility enables bacteria to reach their favorable niche within the host. The human foodborne pathogen Campylobacter jejuni produces two heavily glycosylated structural flagellins (FlaA and FlaB that form the flagellar filament. It also encodes the non-structural FlaC flagellin which is secreted through the flagellum and has been implicated in host cell invasion. The mechanisms that regulate C. jejuni flagellin biogenesis and guide the proteins to the export apparatus are different from those in most other enteropathogens and are not fully understood. This work demonstrates the importance of the putative flagellar protein FliS in C. jejuni flagella assembly. A constructed fliS knockout strain was non-motile, displayed reduced levels of FlaA/B and FlaC flagellin, and carried severely truncated flagella. Pull-down and Far Western blot assays showed direct interaction of FliS with all three C. jejuni flagellins (FlaA, FlaB, and FlaC. This is in contrast to, the sensor and regulator of intracellular flagellin levels, FliW, which bound to FlaA and FlaB but not to FlaC. The FliS protein but not FliW preferred binding to glycosylated C. jejuni flagellins rather than to their non-glycosylated recombinant counterparts. Mapping of the binding region of FliS and FliW using a set of flagellin fragments showed that the C-terminal subdomain of the flagellin was required for FliS binding, whereas the N-terminal subdomain was essential for FliW binding. The separate binding subdomains required for FliS and FliW, the different substrate specificity, and the differential preference for binding of glycosylated flagellins ensure optimal processing and assembly of the C. jejuni flagellins.

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

    Directory of Open Access Journals (Sweden)

    Kvin Lertpiriyapong

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

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

    Science.gov (United States)

    2012-01-01

    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. PMID:23051923

  4. Antibiotico – resistenza in Campylobacter jejuni /coli in Italia nel corso del 2002: risultati di una indagine policentrica

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    Daniele Crotti

    2003-03-01

    Full Text Available During 2002, inside Campyg Working Group Project, we examined from 8 italian laboratories the patterns of resistances of 205 strains of Campylobacter spp.These strains were: 88% C. jejuni (68% biotype 1, 31% biotype 2, 1% biotype 3, 11% C. coli (95.5% biotype 1 and 1% Clo’s.We observed these resistances: to erythromycin 3.5% (2.2% in C. jejuni, 13.6% in C. coli, to rokitamicin 2.5 % (1.1% and 13.6% respectively, to ciprofloxacin 49.5% (51.1% and 63.6%, to tetracycline 45.0% (43.3% and 59.1%, to minocycline 41.6% (40.0% and 54.5%, to gentamycin 0.5%, to chloramphenicole no ones. Some differences concerning frequency of resistances were observed among 8 laboratories, also between C. jejuni b. 1 and C. jejuni b. 2, but these were not significative. Some suggests are given for continuing this surveillance, from an epidemiological point of view too.

  5. Determination of ciprofloxacin and nalidixic acid resistance in Campylobacter jejuni with a fluorogenic polymerase chain reaction assay.

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    Padungtod, Pawin; Kaneene, John B; Wilson, David L; Bell, Julia; Linz, John E

    2003-02-01

    A fluorogenic polymerase chain reaction assay for the gyrA gene was used to determine the frequency of a Thr-86 mutation in Campylobacter jejuni isolates from food animals and humans in northern Thailand and to investigate the correlation between this mutation and bacterial resistance to fluoroquinolones. Eighty-four isolates of C. jejuni were used: 65 from healthy chickens on farms, 16 from chickens at the slaughterhouse, 1 from chicken meat at the market, and 1 from a healthy farm worker. The microbroth dilution technique was used for in vitro susceptibility testing. MIC breakpoints established by the National Antimicrobial Resistance Monitoring System were used to categorize the resistance of C. jejuni to ciprofloxacin and nalidixic acid. Sixty of the 84 C. jejuni isolates tested carried the Thr-86 mutation in the gyrA gene. All isolates with ciprofloxacin MICs of > or = 2 mg/liter carried the mutation, and no isolates with nalidixic acid MICs of < or = 16 mg/liter carried the Thr-86-to-Ile mutation. There was a very strong association between ciprofloxacin resistance and the presence of the mutation (kappa = 0.971, P < 0.01). The association between the presence of the Thr-86-to-Ile mutation and nalidixic acid resistance was weaker (kappa 0.859: P < or = 0.01).

  6. Level of Campylobacter jejuni from naturally contaminated chicken liver and chicken legs in various task: a cross contamination study

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    New, C.Y.

    2017-03-01

    Full Text Available Cross contamination is one of the most important contributing factors in foodborne illness originating in household environments. The objective of this research was to determine the transfer between naturally contaminated chicken liver and leg to cutting board, hand glove, knife and cucumber, during slicing. The microorganism tested was Campylobacter jejuni and the results showed that the pathogen transferred to all utensils, at different transfer rate, despite the low level of the naturally contaminating pathogen. With unknown concentration bacteria in the naturally contaminated samples, a proportion of the utensils were still contaminated with C. jejuni and not surprisingly, when the sample were contaminated with higher concentrations of the pathogen, a higher proportion of the utensils had detectable C. jejuni cells present, though in many cases cross contamination seems to be a random event. Transfer of the naturally contaminating C. jejuni from the chicken liver and leg to the utensils were <3.0 to 9.2 MPN/g and <3.0 to 3.0 MPN/g, respectively. The results of this study highlighted the potential for cross contamination of food borne pathogens in the kitchen environment.

  7. Presenting a rapid method for detection of Bacillus cereus, Listeria monocytogenes and Campylobacter jejuni in food samples

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    Ali Razei

    2017-08-01

    Full Text Available Objective(s: Listeria monocytogens, Bacillus cereus and Campylobacter jejuni are three toxin producing bacteria over the world, especially in Iran, and it is essential to find a certain, rapid procedure to identify these microorganisms. In this research, these bacteria were simultaneously detected by multiplex PCR technique in foods. Materials and Methods: The primary approval of bacterial strains was performed by biochemical tests. PCR primers were designed based on the nucleotide sequences of the NHEB/NHEC gene of B. cereus, the hly gene of L. monocytogenes and the C gene of C. jejuni. The specificity of Multiplex PCR method was determined using seven food poisoning bacteria including Salmonella typhi, Shigella dysentery, Yersinia pestis, Staphylococcus aureus, Clostridium perfringens, Clostridium botulinum and Vibrio cholerae. To confirm the reaction, DNA extraction was performed from 30 food samples (milk, and gene amplification was performed by PCR. The length of amplified fragments was 300 bp, 210 bp and 160 bpfor NHEB/NHEC, hly and C genes, respectively. Results: The detection limits of the PCR method were 5, 4 and 3 pg for L. monocytogenes, B. cereus and C. jejuni, respectively. Specifisity test showed that this reaction is spesific to these 3 bacteria. Conclusion: In this study, we  introduced a new multiplex PCR method for simultsnus detection of L. monocytogens, B. cereus and C. jejuni. These results can be used  for detection of other toxin producing bacteria in food.

  8. Presenting a rapid method for detection of Bacillus cereus, Listeria monocytogenes and Campylobacter jejuni in food samples.

    Science.gov (United States)

    Razei, Ali; Sorouri, Rahim; Mousavi, Seyed Latif; Nazarian, Shahram; Amani, Jafar; Aghamollaei, Hosien

    2017-09-01

    Listeria monocytogens, Bacillus cereus and Campylobacter jejuni are three toxin producing bacteria over the world, especially in Iran, and it is essential to find a certain, rapid procedure to identify these microorganisms. In this research, these bacteria were simultaneously detected by multiplex PCR technique in foods. The primary approval of bacterial strains was performed by biochemical tests. PCR primers were designed based on the nucleotide sequences of the NHEB/NHEC gene of B . cereus , the hly gene of L. monocytogenes and the C gene of C. jejuni . The specificity of Multiplex PCR method was determined using seven food poisoning bacteria including Salmonella typhi, Shigella dysentery, Yersinia pestis, Staphylococcus aureus, Clostridium perfringens, Clostridium botulinum and Vibrio cholerae . To confirm the reaction, DNA extraction was performed from 30 food samples (milk), and gene amplification was performed by PCR. The length of amplified fragments was 300 bp, 210 bp and 160 bp for NHEB/NHEC , hly and C genes, respectively. The detection limits of the PCR method were 5, 4 and 3 pg for L. monocytogenes , B. cereus and C. jejuni , respectively. Specifisity test showed that this reaction is spesific to these 3 bacteria. In this study, we introduced a new multiplex PCR method for simultsnus detection of L. monocytogens, B. cereus and C. jejuni . These results can be used for detection of other toxin producing bacteria in food.

  9. Application of protective cultures against Listeria monocytogenes and Campylobacter jejuni in chicken products packaged under modified atmosphere.

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    Melero, B; Vinuesa, R; Diez, A M; Jaime, I; Rovira, J

    2013-04-01

    To control the growth, or reduce the numbers, of food pathogens such as Listeria monocytogenes and Campylobacter jejuni in chicken products packaged under modified atmosphere (MAP), the effectiveness of protective cultures was evaluated in this study. Leuconostoc pseudomesenteroides PCK18 reduced the counts of L. monocytogenes by 1.22 log cfu/g in chicken burgers under MAP after 24 d. Furthermore, a reduction of 1.16 log cfu/g in C. jejuni together with a delay in the growth of lactic acid bacteria was obtained in chicken legs inoculated with Bifidobacterium longum ssp. longum PCB133 and packaged under MAP after 9 d. The combination and concentration of gases in the MAP doubled the products' shelf-life in comparison with air-packaged samples in both experiments. In conclusion, this study has shown the effectiveness of 2 protective culture strains against 2 foodborne pathogens, resulting in safer products with a longer shelf-life.