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Sample records for thermotolerant campylobacters assay

  1. Toward an international standard for PCR-based detection of food-borne thermotolerant Campylobacters: Assay development and analytical validation

    DEFF Research Database (Denmark)

    Lübeck, Peter Stephensen; Wolffs, P.; On, Stephen L.W.

    2003-01-01

    As part of a European research project (FOOD-PCR), we developed a standardized and robust PCR detection assay specific for the three most frequently reported food-borne pathogenic Campylobacter species, C. jejuni, C. coli, and C. lari. Fifteen published and unpublished PCR primers targeting the 16S...... carcass rinse, unlike both Taq DNA polymerase and DyNAzyme. Based on these results, Tth was selected as the most suitable enzyme for the assay. The standardized PCR test described shows potential for use in large-scale screening programs for food-borne Campylobacter species under the assay conditions....... The inclusivity and exclusivity were 100 and 97%, respectively. In an attempt to find a thermostable DNA polymerase more resistant than Taq to PCR inhibitors present in chicken samples, three DNA polymerases were evaluated. The DNA polymerase Tth was not inhibited at a concentration of 2% (vol/vol) chicken...

  2. Toward an International Standard for PCR-Based Detection of Food-Borne Thermotolerant Campylobacters: Assay Development and Analytical Validation

    OpenAIRE

    Lübeck, P. S.; Wolffs, P.; On, S. L. W.; Ahrens, P.; Rådström, P.; Hoorfar, J.

    2003-01-01

    As part of a European research project (FOOD-PCR), we developed a standardized and robust PCR detection assay specific for the three most frequently reported food-borne pathogenic Campylobacter species, C. jejuni, C. coli, and C. lari. Fifteen published and unpublished PCR primers targeting the 16S rRNA gene were tested in all possible pairwise combinations, as well as two published primers targeting the 23S rRNA gene. A panel of 150 strains including target an...

  3. Development of a monoclonal antibody-based colony blot immunoassay for detection of thermotolerant Campylobacter species.

    Science.gov (United States)

    Huang, Hongsheng; Phipps-Todd, Beverley; McMahon, Tanis; Elmgren, Catherine L; Lutze-Wallace, Cheryl; Todd, Zoe A; Garcia, Manuel M

    2016-11-01

    Campylobacter species, particularly thermotolerant Campylobacter spp., such as C. jejuni, are major human foodborne pathogens. Culture methods have been routinely used for the detection of this organism in various types of samples. An alternative, simple and rapid confirmation test(s) without further tedious biochemical tests would be useful. Meanwhile, Campylobacter-like colonies can be difficult to identify on agar plates overgrown with competitive bacteria, which can lead to false-negative results. This study was to develop a simple colony blot immunoassay using a new monoclonal antibody (Mab) produced in the present study for rapid screening, confirmation and quantification of campylobacters on culture agar plates. The procedure developed in this study was able to specifically detect thermotolerant Campylobacter spp., but not other non-thermotolerant Campylobacter and non-Campylobacter reference strains tested. This assay could detect 10 5 cells in a single dot. This assay showed 100% correlation with the culture method for the blotted membranes from 21 either chicken meat or vegetable samples experimentally inoculated with thermotolerant campylobacters. Among 101 natural samples of chicken meat (n=44), chicken feces (n=20) and vegetables (n=37), this assay also showed positive for 23 chicken meat and 14 fecal samples that were positive for thermotolerant campylobacters by culture method, and identified four additional suspects that were culture negative. Membranes stored at 4°C for at least 4years could also be used for this assay. The assay developed in this study can be used in quantitative study for immediate or archival usage, and for diagnostic test to preliminarily confirm the presence of thermotolerant Campylobacter on agar plates. Crown Copyright © 2016. Published by Elsevier B.V. All rights reserved.

  4. Towards an international standard for PCR-based detection of food-borne thermotolerant Campylobacters: assay development and analytical validation.

    OpenAIRE

    Lübeck, P S; Wolffs, Petra; On, S L; Ahrens, P; Rådström, Peter; Hoorfar, J

    2003-01-01

    As part of a European research project (FOOD-PCR), we developed a standardized and robust PCR detection assay specific for the three most frequently reported food-borne pathogenic Campylobacter species, C. jejuni, C. coli, and C. lari. Fifteen published and unpublished PCR primers targeting the 16S rRNA gene were tested in all possible pairwise combinations, as well as two published primers targeting the 23S rRNA gene. A panel of 150 strains including target and nontarget strains was used in ...

  5. The prevalence of thermotolerant Campylobacter species in food ...

    African Journals Online (AJOL)

    Background: Thermotolerant Campylobacter spp. is known to occur in the intestinal systems of a wide variety of domestic and wild animals. Although Campylobacter jejuni and Campylobacter coli cause acute diarrhoeal diseases in humans worldwide, they mostly manifest themselves in an apparently healthy carrier state in ...

  6. The prevalence of thermotolerant Campylobacter species in food ...

    African Journals Online (AJOL)

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    Abstract. Background: Thermotolerant Campylobacter spp. is known to occur in the intestinal systems of a wide variety of domestic and wild animals. Although Campylobacter jejuni and Campylobacter coli cause acute diarrhoeal diseases in humans worldwide, they mostly manifest themselves in an apparently healthy ...

  7. Classical and Molecular Identification of Thermotolerant Campylobacters from Poultry Meat

    Directory of Open Access Journals (Sweden)

    Tina Zorman

    2002-01-01

    Full Text Available Poultry meat samples from Slovenian retail market were examined for the presence of thermotolerant campylobacters. The isolates were identified by phenotypic and genotypic methods. ISO 10272 recommendations were followed for phenotypic identification. Different PCR assays, targeting species specific DNA regions in C. jejuni and C. coli, were checked for their applicability in identification. High degree of tested samples was positive (27/33, with significant proportion of C. coli (32 % among identified strains. High percentage of C. jejuni strains (54 % were hippurate negative. Phenotypic identification was therefore found to be inconvenient because of the presence of the strains with atypical phenotype and possible misinterpretation of test results. Multiplex PCR, targeting hippuricase gene in C. jejuni and species specific region in C. coli, was found to be an efficient method that allowed fast, simple and accurate identification of C. jejuni and C. coli. FlaA PCR is a reliable method to identify the group C. jejuni/C. coli, but it does not differentiate between the two species. CdtB PCR is inconvenient because of many false negative and some false positive results.

  8. 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

  9. Toward an international standard for PCR-based detection of food-borne thermotolerant campylobacters: Validation in a multicenter collaborative trial

    DEFF Research Database (Denmark)

    Lübeck, Peter Stephensen; Cook, N.; Wagner, M.

    2003-01-01

    As part of a European research project, the performance of a PCR assay to detect food-borne thermotolerant campylobacters (Campylobacter jejuni, C. coli, and C. lari) was evaluated through an international collaborative trial involving 12 participating laboratories. DNA from 10 target and 8...

  10. Human risk from thermotolerant Campylobacter on broiler meat in Denmark

    DEFF Research Database (Denmark)

    Boysen, Louise; Nauta, Maarten; Ribeiro Duarte, Ana Sofia

    2013-01-01

    This paper describes a new approach by which changes over time in the relative risk of human campylobacteriosis from broiler meat are evaluated through quantitative microbiological risk assessment modelling. Danish surveillance data collected at retail from 2001 to 2010 on numbers of thermotolerant...... Campylobacter spp. on Danish produced and imported chilled and frozen broiler meat were the basis for the investigation. The aim was to explore if the risk from the different meat categories had changed over time as a consequence of implemented intervention strategies. The results showed a slight decrease from...... 2005 to 2008 in the human risk from Danish produced broiler meat, and a decrease from 2005 to 2010 in the risk from imported chilled meat. This risk reduction coincides with control measures implemented to reduce Campylobacter in Danish and imported chilled broiler meat. The human risk...

  11. An immunomagnetic separation/loop-mediated isothermal amplification method for rapid direct detection of thermotolerant Campylobacter spp. during poultry production.

    Science.gov (United States)

    Romero, M R; D'Agostino, M; Arias, A Pérez; Robles, S; Casado, C Fernández; Iturbe, L Orueta; Lerma, O Gurrutxaga; Andreou, M; Cook, N

    2016-02-01

    To develop a rapid test for thermotolerant Campylobacter in poultry faeces. The reported method is based on immunomagnetic separation and loop-mediated isothermal DNA amplification (IMS/LAMP). This LAMP assay is specific (demonstrated using 10 Campylobacter strains and 13 non-Campylobacter bacterial species) and sensitive (95% probability of detecting 22 genome copies). A competitive internal amplification control (IAC) has been incorporated to give unambiguous determination of negative results. Immunoseparation of Campylobacter allows direct LAMP detection from poultry boot swab samples in 90 min without enrichment or DNA purification (74% probability of detecting 10(4) CFU ml(-1) of a boot swab suspension). The analysis of 17 samples from commercial turkey farms showed 100% correlation with parallel results obtained by standard microbiological methods. A rapid test has been developed for direct detection of thermotolerant Campylobacter spp. in boot swab samples, thus bypassing culture enrichment or DNA extraction. The test has potential to be carried out by farm personnel on site. The method offers an inexpensive approach to monitor poultry infection in near real time, assisting flock management and controls to prevent introduction of Campylobacter into the food chain. © 2015 The Society for Applied Microbiology.

  12. Isolation of thermotolerant campylobacters and C. hyointestinalis from rectal swabs of healthy pigs

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    Mrenoski Slavco

    2007-11-01

    Full Text Available Thermotolerant campylobacters are the most common bacterial etiological agents of human infectious gastroenteritis worldwide. The most frequent isolated species among them are Campylobacter jejuni and C. coli, and less frequent C. upsaliensis and C. lari. Also C. hyointestinalis, that not belong to the group of thermotolerant campylobacters, has been indicate as an agent of human infectious gastroenteritis. Natural reservoir of all named campylobacters is the intestinal tract of many mammals and birds, including domestic animals. In these animals, campylobacters are commonly present as commensals and their feces is considered as a prime source for environmental contamination. Unlike the human feces which is usually examined in the cases of diarrhea, thermotolerant campylobacters and C. hyointestinalis in the animal feces are generally present in a much lesser amount and the isolation very often could be unsuccessful. The aim of this study was to estimate the validity of applied procedure for isolation (and identification of thermotolerant campylobacters and C. hyointestinalis from pig rectal swabs, as a procedure for detection of healthy animal carriers.

  13. Thermotolerant Coliforms Are Not a Good Surrogate for Campylobacter spp. in Environmental Water ▿

    Science.gov (United States)

    St-Pierre, Karen; Lévesque, Simon; Frost, Eric; Carrier, Nathalie; Arbeit, Robert D.; Michaud, Sophie

    2009-01-01

    This study aimed to assess the importance of quantitatively detecting Campylobacter spp. in environmental surface water. The prevalence and the quantity of Campylobacter spp., thermotolerant coliforms, and Escherichia coli in 2,471 samples collected weekly, over a 2-year period, from 13 rivers and 12 streams in the Eastern Townships, Québec, Canada, were determined. Overall, 1,071 (43%), 1,481 (60%), and 1,463 (59%) samples were positive for Campylobacter spp., thermotolerant coliforms, and E. coli, respectively. There were weak correlations between the weekly distributions of Campylobacter spp. and thermotolerant coliforms (Spearman's ρ coefficient = 0.27; P = 0.008) and between the quantitative levels of the two classes of organisms (Kendall tau-b correlation coefficient = 0.233; P coliforms. These findings suggest that microbial monitoring of raw water by using only fecal indicator organisms is not sufficient for assessing the occurrence or the load of thermophilic Campylobacter spp. Insights into the role of environmental water as sources for sporadic Campylobacter infection will require genus-specific monitoring techniques. PMID:19734335

  14. Thermotolerant coliforms are not a good surrogate for Campylobacter spp. in environmental water.

    Science.gov (United States)

    St-Pierre, Karen; Lévesque, Simon; Frost, Eric; Carrier, Nathalie; Arbeit, Robert D; Michaud, Sophie

    2009-11-01

    This study aimed to assess the importance of quantitatively detecting Campylobacter spp. in environmental surface water. The prevalence and the quantity of Campylobacter spp., thermotolerant coliforms, and Escherichia coli in 2,471 samples collected weekly, over a 2-year period, from 13 rivers and 12 streams in the Eastern Townships, Québec, Canada, were determined. Overall, 1,071 (43%), 1,481 (60%), and 1,463 (59%) samples were positive for Campylobacter spp., thermotolerant coliforms, and E. coli, respectively. There were weak correlations between the weekly distributions of Campylobacter spp. and thermotolerant coliforms (Spearman's rho coefficient = 0.27; P = 0.008) and between the quantitative levels of the two classes of organisms (Kendall tau-b correlation coefficient = 0.233; P coliforms. These findings suggest that microbial monitoring of raw water by using only fecal indicator organisms is not sufficient for assessing the occurrence or the load of thermophilic Campylobacter spp. Insights into the role of environmental water as sources for sporadic Campylobacter infection will require genus-specific monitoring techniques.

  15. 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

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    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.

  16. The effect of slaughter operations on the contamination of chicken carcasses with thermotolerant Campylobacter

    DEFF Research Database (Denmark)

    Rosenquist, Hanne; Sommer, Helle Mølgaard; Nielsen, Niels L.

    2006-01-01

    To evaluate the effect of specific slaughter operations on the contamination of broiler carcasses with naturally occurring thermotolerant Campylobacter, experiments were carried out in two Danish commercial slaughter plants (Plant I and Plant 11). Six broiler flocks determined Campylobacter...... concentration of 0.5 log(10) cfu/g in average, whereas no significant changes were observed during this operation in Plant II. Air chilling (Plant 1) and water chilling (Plant 11), both including a carcass wash prior to the chilling operation, caused similar, but significant reductions of 0.83 and 0.97 log(10......) cfu/g, respectively. In packed frozen chickens (Plant II) an additional reduction of 1.38 log(10) cfu/g in average was obtained due to the freezing operation. In packed chilled chickens (Plant 1), however, the number of thermotolerant Campylobacter per gram remained at the same level as after air...

  17. Prevalence and Antibiotic Resistance of Thermotolerant Campylobacter spp. in Chicken Meat Sold in İstanbul

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    Serkan Kemal BÜYÜKÜNAL

    2017-07-01

    Full Text Available Campylobacter spp. are some of the most common causes of bacterial diarrhea in humans worldwide. They are mainly considered as foodborne pathogens that are found in raw or undercooked poultry and serve as an important source of sporadic campylobacteriosis. The present study was aimed to determine the prevalence and the antimicrobial resistance patterns of thermotolerant Campylobacter spp. in chicken meat. A total of 176 samples of chicken meat were analyzed using PCM and BAX® system. The samples analyzed included: 56 samples of whole chicken, 27 samples of chicken breast, 33 samples of chicken thigh, 25 samples of chicken drumstick and 35 samples of chicken wings. Samples of all the fresh chicken meat sold in İstanbul were randomly purchased from different major supermarkets in their original, individual packages. Laboratory analyses to detect thermotolerant Campylobacter spp. were performed in accordance with the ISO 10272-1, 2006 standard (qualitative analysis. API® Campy (BioMerieux, Marcy-l’Etoile, France was used for the confirmation of presumptive colonies. Campylobacter isolates were subjected to antimicrobial susceptibility tests by the disc diffusion method as recommended by the National Committee for Clinical Laboratory Standards. Zones of growth inhibition were evaluated according to the NCCLS standards. Using PCM, the prevalence of C. coli, C. jejuni and C. lari was determined as 15.34, 8.52 and 1.7%, respectively. However, using BAX® system, the prevalence was determined as 15.90, 18.75 and 1.7% for C. coli, C. jejuni and C. lari, respectively. C. coli was resistant to nalidixic acid (78.57%, ofloxacin (14.29% norfloxacin (10.71% and ampicillin (10.71%. But the highest resistance was observed to nalidixic acid (90.91% for C. jejuni and (100% for C. lari. In conclusion, considering the public health, chicken meat is a common source for Campylobacter strains and antibiotics should be used carefully in veterinary medicine.

  18. Occurrence of thermotolerant Campylobacter spp. at different stages of the poultry meat supply chain in Argentina.

    Science.gov (United States)

    Zbrun, M V; Romero-Scharpen, A; Olivero, C; Rossler, E; Soto, L P; Rosmini, M R; Sequeira, G J; Signorini, M L; Frizzo, L S

    2013-11-01

    The objectives of this study were to investigate the occurrence and concentration of thermotolerant Campylobacter spp. at different stages of the poultry meat supply chain in Argentina. Three integrated poultry companies were sampled. Each supply chain was considered at different stages from the reproductive farm to chicken meat at a retail market. The stages sampled were: (a) hens from breeder flocks, (b) eggs in the incubator, (c) broiler chickens in flocks (aged 5 weeks), (d) chickens at a slaughterhouse, and (e) chicken meat at a retail market. The chickens sampled along each supply chain were in the same batch. Samples collected were: (a) cloacal samples from hens and chickens on the farms, (b) fertile eggs, (c) feed, water and litter from flocks, (d) chicken carcasses from the slaughterhouse and retail market, and (e) caeca and livers from the slaughterhouse. Samples obtained were examined for Campylobacter spp. The isolates were biotyped and the genus and species identified by PCR. Campylobacter spp. on chicken carcasses at slaughterhouse and retail market were enumerated. The highest proportions of Campylobacter positive samples were observed in carcasses at retail (25/30, 83.3%) and faecal samples from breeding hens (27/45, 60.0%). Only 3.3% (3/90) samples collected from broiler chickens aged meat in Argentina. The proportions of Campylobacter-positive samples found in this preliminary study indicate that a large proportion of the cases of human gastroenteritis in Argentina may be due to this pathogen. Human cases of gastroenteritis should be studied in greater detail and measures should be developed to reduce the proportion of poultry products that are contaminated by Campylobacter species.

  19. A LightCycler real-time PCR hybridization probe assay for detecting food-borne thermophilic Campylobacter.

    Science.gov (United States)

    Perelle, Sylvie; Josefsen, Mathilde; Hoorfar, Jeffrey; Dilasser, Françoise; Grout, Joël; Fach, Patrick

    2004-10-01

    In a previous study, we reported the performance of a PCR assay amplifying 287-bp of the 16S rRNA gene of thermo-tolerant Campylobacter (C. jejuni, C. lari, C. coli) through an international ring-trial involving 12 participating laboratories. Based on the validated set of primers, a LightCycler real-time PCR assay (LC-PCR), which used fluorescent hybridization probes was developed. The test incorporated an internal amplification control co-amplified with the 16S rRNA gene of Campylobacter to monitor potential PCR inhibitors and ensure successful amplifications. The specificity study involving 39 Campylobacter and nine strains of other species indicated that the LC-PCR test was highly specific, giving cross-reactivity with only one strain of C. upsaliensis (CCUG19559). The sensitivity of the LC-PCR assay, evaluated in 32 spiked poultry-rinse or pork carcass-swab samples, was determined at 10CFU/ml carcass-rinse. The prevalence of samples positive for thermo-tolerant Campylobacter was 58.8% in 68 naturally contaminated poultry rinse samples tested by LC-PCR and the data were in good concordance with those of bacteriological method. The Ct values of the three replicates obtained for each sample tested in three different runs demonstrate that the LC-PCR was highly reproducible and afford a powerful tool for rapid detection of the thermo-tolerant Campylobacter strains.

  20. Validation of a PCR-based method for detection of food-borne thermotolerant Campylobacters in a multicenter collaborative trial

    DEFF Research Database (Denmark)

    Josefsen, Mathilde Hartmann; Cook, N.; D'Agostino, M.

    2004-01-01

    A PCR-based method for rapid detection of food-borne thermotolerant campylobacters was evaluated through a collaborative trial with 12 laboratories testing spiked carcass rinse samples. The method showed an interlaboratory diagnostic sensitivity of 96.7% and a diagnostic specificity of 100...

  1. A LightCycler real-time PCR hybridization probe assay for detecting food-borne thermophilic Campylobacter

    DEFF Research Database (Denmark)

    Perelle, S.; Josefsen, Mathilde Hartmann; Hoorfar, Jeffrey

    2004-01-01

    In a previous study, we reported the performance of a PCR assay amplifying 287-bp of the 16S rRNA gene of thermo-tolerant Campylobacter (C. jejuni, C. lari, C. coli) through an international ring-trial involving 12 participating laboratories. Based on the validated set of primers, a LightCycler r...... for each sample tested in three different runs demonstrate that the LC-PCR was highly reproducible and afford a powerful tool for rapid detection of the thermotolerant Campylobacter strains.......In a previous study, we reported the performance of a PCR assay amplifying 287-bp of the 16S rRNA gene of thermo-tolerant Campylobacter (C. jejuni, C. lari, C. coli) through an international ring-trial involving 12 participating laboratories. Based on the validated set of primers, a Light......Cycler real-time PCR assay (LC-PCR), which used fluorescent hybridization probes was developed. The test incorporated an internal amplification control co-amplified with the 16S rRNA gene of Campylobacter to monitor potential PCR inhibitors and ensure successful amplifications. The specificity study involving...

  2. Towards an international standard for PCR-based detection of foodborne thermotolerant campylobacters: interaction of enrichment media and pre-PCR treatment on carcass rinse samples

    DEFF Research Database (Denmark)

    Josefsen, Mathilde Hartmann; Lübeck, Peter Stephensen; Hansen, F.

    2004-01-01

    As part of a large EU project for standardisation of polymerase chain reaction (PCR), a systematic evaluation of the interaction of enrichment media, type of DNA polymerase and pre-PCR sample treatment for a PCR detecting thermotolerant campylobacters was carried out. The growth-supporting capacity...... and PCR compatibility of enrichment in Preston, Mueller-Hinton and Bolton broth (blood-containing and blood-free) were evaluated. The effect of resin-based DNA extraction and DNA extraction by boiling on the final PCR assay was investigated. The time-course studies indicated that a 20-h sample enrichment...... in blood-containing Bolton broth, followed by a simple resin-based extraction of DNA and a PCR amplification using Tth polymerase, resulted in strong and clear PCR amplicons for target (287 bp) and internal amplification control (IAC, 124 bp). The enrichment PCR-based method, tested on 68 presumably...

  3. 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...

  4. Reduction of thermotolerant Campylobacter species on broiler carcasses following physical decontamination at slaughter

    DEFF Research Database (Denmark)

    Boysen, Louise; Rosenquist, Hanne

    2009-01-01

    To reduce the incidences of human Campylobacter infections, a number of countries are investigating methods for reducing human exposure to Campylobacter from broiler meat. In addition to implementing biosecurity measures at the farm, Campylobacter may be controlled by reducing Campylobacter counts...

  5. Enhanced enrichment and detection of thermotolerant Campylobacter species from water using the Portable Microbe Enrichment Unit and real-time PCR.

    Science.gov (United States)

    Pitkänen, Tarja; Bräcker, Juliane; Miettinen, Ilkka T; Heitto, Anneli; Pesola, Jouni; Hakalehto, Elias

    2009-07-01

    An enhanced enrichment using the Portable Microbe Enrichment Unit (PMEU) with the microaerobic bubbling of broths was applied for the detection of thermotolerant Campylobacter species from water. This PMEU enrichment was compared with the conventional static enrichment of the international standard ISO 17995:2005. In addition, Campylobacter detection after enrichment using a real-time PCR detection was compared with colony counts. The tests with stressed Campylobacter jejuni cells in drinking water indicated that the PMEU enrichment yielded a significantly higher number of Campylobacter cells in the Bolton broth compared with the conventional static incubation. Application of the real-time PCR technique shortened the Campylobacter detection time. This combination of method modifications can be used for Campylobacter detection from water and adds methodological repertoire for the rapid survey and management of waterborne outbreaks.

  6. Reduction of thermotolerant Campylobacter species on broiler carcasses following physical decontamination at slaughter

    DEFF Research Database (Denmark)

    Boysen, Louise; Rosenquist, Hanne

    2009-01-01

    through physical decontamination of the meat. The current study was conducted to compare the Campylobacter-reducing ability of three physical decontamination techniques, forced air chilling, crust freezing, and steam-ultrasound, performed in the plant with naturally contaminated broiler chickens......To reduce the incidences of human Campylobacter infections, a number of countries are investigating methods for reducing human exposure to Campylobacter from broiler meat. In addition to implementing biosecurity measures at the farm, Campylobacter may be controlled by reducing Campylobacter counts...... in an increase of 0.9 log CFU per carcass, suggesting that Campylobacter counts also may be reduced by optimizing the hygienic design of equipment or by physical removal of fecal contamination....

  7. Detection of Resistance to Macrolides in Thermotolerant Campylobacter Species by Fluorescence In Situ Hybridization▿

    OpenAIRE

    Haas, Michaela; Essig, Andreas; Bartelt, Edda; Poppert, Sven

    2008-01-01

    The resistance of enteritis-causing Campylobacter strains to erythromycin is an emerging problem. We therefore evaluated fluorescence in situ hybridization (FISH) for the rapid detection of resistance using 74 campylobacter isolates. FISH showed specificity and sensitivity of 100% for the detection of high-level resistance.

  8. Seasonal influence on the prevalence of thermotolerant Campylobacter in retail broiler meat in Denmark

    DEFF Research Database (Denmark)

    Boysen, Louise; Vigre, Håkan; Rosenquist, Hanne

    2011-01-01

    In Denmark, the incidence of human campylobacteriosis cases, as well as the Campylobacter prevalence in broiler flocks, is strongly influenced by season with a summer peak in July–August. Therefore, it was considered that the prevalence of Campylobacter in broiler meat sold at retail in Denmark...

  9. Aislamiento de especies termotolerantes de Campylobacter en dos poblaciones de pollos criados con y sin confinamiento Isolation of thermotolerant Campylobacter species from two populations of chickens bred in confinement and at liberty

    Directory of Open Access Journals (Sweden)

    Alvaro Tresierra-Ayala

    1995-10-01

    Full Text Available Se estudió la frecuencia del aislamiento de Campylobacter spp. en pollos domésticos y pollos mantenidos en confinamiento permanente, en la ciudad de Iquitos (Perú. Campylobacter spp. fue aislado en 54,0% en el primer grupo y 35,0% en el segundo (pEstudou-se a freqüência de isolamento de Campylobacter spp. em frangos domésticos e frangos mantidos em confinamento permanente, na cidade de lquitos (Peru. Campylobacter spp. foi isolado em 54,0% no primeiro grupo e 35,0% no segundo (pThe isolation rates of thermotolerant Campylobacter species in free-ranging domestic chickens and confined chickens from Iquitos city, Peru, were determined. Campylobacter spp. were isolated in 54,0% of the former group of chickens, being less frequent (35,0% in the latter (p<0,05. Of the classical thermotolerant species, C. jejuni and C. coli were the most frequent. However, the presence C. lari suggests that the chickens might be an important reservoir of this bacterium.

  10. [Cultural detection of thermotolerant Campylobacter spp. in food--potentials and limitations of diagnostic tools in the context of official food control].

    Science.gov (United States)

    Messelhäusser, Ute; Thärigen, Diana; Fella, Christiane; Schreiner, Hermann; Busch, Ulrich; Höller, Christiane

    2015-01-01

    Thermotolerant Campylobacter spp. rank among the most important foodborne pathogens in Germany. Therefore a necessity for rapid and routinely useable detection methods exists also in the area of food microbiology. A reliable, cultura qualitative, but also quantitative detection of thermotolerant Campylobacter spp. pose a challenge, at least concerning special food matrices, especially because in the context of official food control the cultural detection of thermotolerant Campylobacter spp. is needed. This was the reason, why different cultural detection methods, beside the standard procedure of ISO 10272:2006, in combination with molecular and immunological screening methods were tested at the Bavarian Health and Food Safety Authority (LGL) during the last years for the use in routine diagnostic using different food matrices of animal and plant origin. The results of the comparative studies showed clearly that no enrichment broth tested gave completely satisfactory results for an only culture-based detection the combination with a screening method is therefore recommended for a rapid and reliable detection. But in this case the user should take into account that the sensitivity of such molecular and immunological methods is normally so high that in some cases, depending on the food matrix and processing step, the isolation of the pathogen would not be possible in samples, which were positive in the screening methods.

  11. Towards an international standard for PCR-based detection of foodborne thermotolerant campylobacters: interaction of enrichment media and pre-PCR treatment on carcass rinse samples.

    Science.gov (United States)

    Josefsen, M H; Lübeck, P S; Hansen, F; Hoorfar, J

    2004-07-01

    As part of a large EU project for standardisation of polymerase chain reaction (PCR), a systematic evaluation of the interaction of enrichment media, type of DNA polymerase and pre-PCR sample treatment for a PCR detecting thermotolerant campylobacters was carried out. The growth-supporting capacity and PCR compatibility of enrichment in Preston, Mueller-Hinton and Bolton broth (blood-containing and blood-free) were evaluated. The effect of resin-based DNA extraction and DNA extraction by boiling on the final PCR assay was investigated. The time-course studies indicated that a 20-h sample enrichment in blood-containing Bolton broth, followed by a simple resin-based extraction of DNA and a PCR amplification using Tth polymerase, resulted in strong and clear PCR amplicons for target (287 bp) and internal amplification control (IAC, 124 bp). The enrichment PCR-based method, tested on 68 presumably naturally contaminated poultry-rinse samples, showed a diagnostic sensitivity of 97.5% (39 PCR-positive/40 total positive samples) and a diagnostic specificity of 100% (28 PCR-negative/28 total negative samples; P=0.32) when compared to a standard bacteriological method (ISO 10272). Copyright 2004 Elsevier B.V.

  12. THERMOTOLERANT Campylobacter SPECIES ISOLATED FROM PSITTACIFORMES IN THE PERUVIAN AMAZON REGION

    Directory of Open Access Journals (Sweden)

    Alvaro TRESIERRA-AYALA

    1998-07-01

    Full Text Available Foi determinada a freqüência de isolamento de campylobacters termotolerantes em Psittaciformes silvestres capturados na região amazônica do Peru. Campylobacters foram isolados em 10/142 (7.0% dos animais estudados, sendo C. jejuni subsp. jejuni biovar I (6/10 o mais freqüente, seguido de C. coli biovar II (2/10, C. lari não foi isolado. Os resultados sugerem que estas aves podem ser importantes reservatórios destas bactérias.

  13. [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.

  14. Development of a PCR assay suitable for Campylobacter spp. mass screening programs in broiler production

    DEFF Research Database (Denmark)

    Bang, Dang Duong; Pedersen, Karl; Madsen, Mogens

    2001-01-01

    culture techniques since 1998. However, using conventional culture methods is time consuming and laborious, and therefore a Polymerase Chain Reaction (PCR) Campylobacter detection assay suitable for mass screening of cloacal swab samples from broilers was developed. By comparing the PCR detection...... with conventional culture methods, significantly more samples were found positive for Campylobacter with the PCR method. The PCR method is rapid, sensitive and suitable for mass screening for Campylobacter in poultry. Using this PCR method Campylobacter can be detected within 15 h. Notably, the method can...... be applied to detect Campylobacter directly from chicken feces at the species level....

  15. Evaluation of real-time PCR assays and standard curve optimisation for enhanced accuracy in quantification of Campylobacter environmental water isolates.

    Science.gov (United States)

    Gosselin-Théberge, Maxime; Taboada, Eduardo; Guy, Rebecca A

    2016-10-01

    Campylobacter is a major public health and economic burden in developed and developing countries. This study evaluated published real-time PCR (qPCR) assays for detection of Campylobacter to enable selection of the best assays for quantification of C. spp. and C. jejuni in environmental water samples. A total of 9 assays were compared: three for thermotolerant C. spp. targeting the 16S rRNA and six for C. jejuni targeting different genes. These assays were tested in the wet-lab for specificity and sensitivity against a collection of 60, genetically diverse, Campylobacter isolates from environmental water. All three qPCR assays targeting C. spp. were positive when tested against the 60 isolates, whereas, assays targeting C. jejuni differed among each other in terms of specificity and sensitivity. Three C. jejuni-specific assays that demonstrated good specificity and sensitivity when tested in the wet-lab showed concordant results with in silico-predicted results obtained against a set of 211 C. jejuni and C. coli genome sequences. Two of the assays targeting C. spp. and C. jejuni were selected to compare DNA concentration estimation, using spectrophotometry and digital PCR (dPCR), in order to calibrate standard curves (SC) for greater accuracy of qPCR-based quantification. Average differences of 0.56±0.12 and 0.51±0.11 log fold copies were observed between the spectrophotometry-based SC preparation and the dPCR preparation for C. spp. and C. jejuni, respectively, demonstrating an over-estimation of Campylobacter concentration when spectrophotometry was used to calibrate the DNA SCs. Our work showed differences in quantification of aquatic environmental isolates of Campylobacter between qPCR assays and method-specific bias in SC preparation. This study provided an objective analysis of qPCR assays targeting Campylobacter in the literature and provides a framework for evaluating novel assays. Crown Copyright © 2016. Published by Elsevier B.V. All rights reserved.

  16. Enrichment followed by quantitative PCR both for rapid detection and as a tool for quantitative risk assessment of food-borne thermotolerant campylobacters

    DEFF Research Database (Denmark)

    Josefsen, Mathilde Hartmann; Jacobsen, N. R.; Hoorfar, Jeffrey

    2004-01-01

    As part of a large international project for standardization of PCR (Food-PCR; www.pcr.dk), a multiplex, multiplatform, ready-to-go enrichment followed by a real-time PCR method, including an internal amplification control, was developed for detection of food-borne thermotolerant campylobacters...... Organization (ISO)-based culture method by testing low, medium, and high levels of 12 spiked and 66 unspiked, presumably naturally contaminated, chicken rinse samples. In the RotorGene, a positive PCR response was detected in 40 samples of the 66. This was in complete agreement with the enriched ISO culture...... naturally contaminated chicken samples, which indicates PCR's additional potential as a tool for quantitative risk assessment. Signal from the internal amplification control was detected in all culture-negative samples (VIC Ct: 23.1 to 28.1). The method will be taken further and validated...

  17. Evaluation of a PCR/DNA Probe Colorimetric Membrane Assay for Identification of Campylobacter spp. in Human Stool Specimens

    OpenAIRE

    Collins, Evelyn; Glennon, Maura; Hanley, Shirley; Murray, Anne-Marie; Cormican, Martin; Smith, Terry; Maher, Majella

    2001-01-01

    DNA was extracted from 50 human stool specimens using the QIAamp DNA stool minikit. PCR amplification was followed by post-PCR hybridization to DNA probes specific for the Campylobacter genus, Campylobacter jejuni, and Campylobacter coli in a colorimetric membrane assay. Thirty-two of 38 culture-positive specimens were PCR/DNA probe positive for C. jejuni. The assay is rapid and simple and can be applied to stool specimens for the detection of Campylobacter.

  18. 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.

  19. 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.

  20. 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

  1. Preston and Park-Sanders protocols adapted for semi-quantitative isolation of thermotolerant Campylobacter from chicken rinse

    DEFF Research Database (Denmark)

    Josefsen, Mathilde Hartmann; Lübeck, Peter Stephensen; Aalbaek, B.

    2003-01-01

    detection methods. Thus, semi-quantitative detection of thermophilic Campylobacter spp. in 20 naturally contaminated chicken rinse samples was carried out using the two most common standard protocols: Preston and Park-Sanders, as proposed by Nordic Committee on Food Analysis (NMKL) and International...... Standard Organization (ISO), respectively. For both protocols, the chicken rinse samples were prepared in 500 ml buffered peptone water, as recommended in the ISO protocol no. 6887-2. The results indicated that the Preston protocol was superior to the Park-Sanders protocol in supporting growth...

  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. 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...

  4. A collaborative study on a Nordic standard protocol for detection and enumeration of thermotolerant Campylobacter in food (NMKL 119, 3. Ed., 2007)

    DEFF Research Database (Denmark)

    Rosenquist, Hanne; Bengtsson, Anja; Hansen, Tina Beck

    2007-01-01

    countries. The laboratories performed qualitative, semi-quantitative and quantitative analyses on samples of chicken meat, cut lettuce, and milk artificially inoculated with different concentrations including blank duplicates of one strain of Campylobacter coli (SLV-27 1) and one strain of Campylobacter...... Campylobacter in chicken meat....

  5. 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 ...

  6. Development of loop-mediated isothermal amplification and PCR assays for rapid and simple detection of Campylobacter fetus subsp. venerealis.

    Science.gov (United States)

    Yamazaki, Wataru; Taguchi, Masumi; Misawa, Naoaki

    2010-07-01

    Campylobacter fetus is divided into CFV and CFF. Because CFV causes bovine genital campylobacteriosis, differentiation of the two subspecies is essential to the implementation of efficient CFV control and eradication programs. We have developed LAMP and duplex PCR assays for rapid and simple detection of CFV. The LAMP assay correctly detected 7 CFV strains and did not detect 53 CFF, 35 non-fetus Campylobacter and 25 non-Campylobacter strains. The PCR assay successfully differentiated the two subspecies. The LAMP and PCR assays were faster than conventional biochemical assays, requiring for detection less than 50 min and less than 4 hr, respectively, from the beginning of DNA extraction from a single colony on blood agar to final determination. Our LAMP and PCR assays are rapid and practical tools for detection of CFV.

  7. Isolation, identification and differentiation of Campylobacter spp. using multiplex PCR assay from goats in Khartoum State, Sudan.

    Science.gov (United States)

    Elbrissi, Atif; Sabeil, Y A; Khalifa, Khalda A; Enan, Khalid; Khair, Osama M; El Hussein, A M

    2017-03-01

    The aim of this study was to identify and characterize thermophilic Campylobacter species in faecal samples from goats in Khartoum State, Sudan, by application of multiplex polymerase chain reaction. Campylobacteriosis is a zoonotic disease of global concern, and the organisms can be transmitted to human via food, water and through contact with farm animals and pets. There are five clinically related Campylobacter species: Campylobacter jejuni (C. jejuni). Campylobacter coli, Campylobacter lari, Campylobacter upsaliensis and Campylobacter fetus. Conventional cultural methods to diagnose campylobacteriosis are tedious and time consuming. Wide ranges of genes have been reported to be used for PCR-based identification of Campylobacter spp. We used a multiplex PCR assay to simultaneously detect genes from the major five clinically significant Campylobacter spp. The genes selected were hipO (hippuricase) and 23S rRNA from glyA (serine hydroxymethyl transferase) from each of C. jejuni. C. coli, C. lari, and C. upsaliensis; and sapB2 (surface layer protein) from C. fetus subsp. fetus. The assay was used to identify Campylobacter isolates recovered from 336 cultured faecal samples from goats in three localities in Khartoum State. C. coli was the most predominant isolate (234; 69.6%), followed by C. jejuni (19; 5.7%), C. upsaliensis (13; 3.9%), C. fetus subsp. fetus (7; 2.1%) and C. lari (6; 1.8%). Twenty-nine goats showed mixed infection with Campylobacter spp., 21 of which harbored two Campylobacter spp., while eight animals were infected with three species. Ten out of twelve goats that displayed diarrhea harbored C. coli only. C. coli, C. jejuni and C. upsaliensis showed significant variation with localities. The prevalence of C. coli was significantly higher (87; 25.9%) in goats from Omdurman, whereas C. jejuni and C. upsaliensis were significantly higher (11; 3.3%, 9; 2.7%) in goats from Khartoum. The multiplex PCR assay was found to be rapid and easy to perform and

  8. 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.

  9. 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.

  10. 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.

  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. 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.

  13. Genomic analysis of Campylobacter fetus subspecies: identification of candidate virulence determinants and diagnostic assay targets

    Directory of Open Access Journals (Sweden)

    Sanchez Daniel O

    2009-05-01

    Full Text Available Abstract Background Campylobacter fetus subspecies venerealis is the causative agent of bovine genital campylobacteriosis, asymptomatic in bulls the disease is spread to female cattle causing extensive reproductive loss. The microbiological and molecular differentiation of C. fetus subsp. venerealis from C. fetus subsp. fetus is extremely difficult. This study describes the analysis of the available C. fetus subsp. venerealis AZUL-94 strain genome (~75–80% to identify elements exclusively found in C. fetus subsp. venerealis strains as potential diagnostic targets and the characterisation of subspecies virulence genes. Results Eighty Kb of genomic sequence (22 contigs was identified as unique to C. fetus subsp. venerealis AZUL-94 and consisted of type IV secretory pathway components, putative plasmid genes and hypothetical proteins. Of the 9 PCR assays developed to target C. fetus subsp. venerealis type IV secretion system genes, 4 of these were specific for C. fetus subsp. venerealis biovar venerealis and did not detect C. fetus subsp. venerealis biovar intermedius. Two assays were specific for C. fetus subsp. venerealis AZUL-94 strain, with a further single assay specific for the AZUL-94 strain and C. fetus subsp. venerealis biovar intermedius (and not the remaining C. fetus subsp. venerealis biovar venerealis strains tested. C. fetus subsp. fetus and C. fetus subsp. venerealis were found to share most common Campylobacter virulence factors such as SAP, chemotaxis, flagellar biosynthesis, 2-component systems and cytolethal distending toxin subunits (A, B, C. We did not however, identify in C. fetus the full complement of bacterial adherence candidates commonly found in other Campylobacter spp. Conclusion The comparison of the available C. fetus subsp. venerealis genome sequence with the C. fetus subsp. fetus genome identified 80 kb of unique C. fetus subsp. venerealis AZUL94 sequence, with subsequent PCR confirmation demonstrating

  14. 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...

  15. A LightCycler real-time PCR hybridization probe assay for detecting food-borne thermophilic Campylobacter

    DEFF Research Database (Denmark)

    Perelle, S.; Josefsen, Mathilde Hartmann; Hoorfar, Jeffrey

    2004-01-01

    Cycler real-time PCR assay (LC-PCR), which used fluorescent hybridization probes was developed. The test incorporated an internal amplification control co-amplified with the 16S rRNA gene of Campylobacter to monitor potential PCR inhibitors and ensure successful amplifications. The specificity study involving...

  16. A novel real-time PCR assay for quantitative detection of Campylobacter fetus based on ribosomal sequences.

    Science.gov (United States)

    Iraola, Gregorio; Pérez, Ruben; Betancor, Laura; Marandino, Ana; Morsella, Claudia; Méndez, Alejandra; Paolicchi, Fernando; Piccirillo, Alessandra; Tomás, Gonzalo; Velilla, Alejandra; Calleros, Lucía

    2016-12-15

    Campylobacter fetus is a pathogen of major concern for animal and human health. The species shows a great intraspecific variation, with three subspecies: C. fetus subsp. fetus, C. fetus subsp. venerealis, and C. fetus subsp. testudinum. Campylobacter fetus fetus affects a broad range of hosts and induces abortion in sheep and cows. Campylobacter fetus venerealis is restricted to cattle and causes the endemic disease bovine genital campylobacteriosis, which triggers reproductive problems and is responsible for major economic losses. Campylobacter fetus testudinum has been proposed recently based on genetically divergent strains isolated from reptiles and humans. Both C. fetus fetus and C. fetus testudinum are opportunistic pathogens for immune-compromised humans. Biochemical tests remain as the gold standard for identifying C. fetus but the fastidious growing requirements and the lack of reliability and reproducibility of some biochemical tests motivated the development of molecular diagnostic tools. These methods have been successfully tested on bovine isolates but fail to detect some genetically divergent strains isolated from other hosts. The aim of the present study was to develop a highly specific molecular assay to identify and quantify C. fetus strains. We developed a highly sensitive real-time PCR assay that targets a unique region of the 16S rRNA gene. This assay successfully detected all C. fetus strains, including those that were negative for the cstA gene-based assay used as a standard for molecular C. fetus identification. The assay showed high specificity and absence of cross-reactivity with other bacterial species. The analytical testing of the assay was determined using a standard curve. The assay demonstrated a wide dynamic range between 10 2 and 107 genome copies per reaction, and a good reproducibility with small intra- and inter-assay variability. The possibility to characterize samples in a rapid, sensitive and reproducible way makes this assay

  17. 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.

  18. Development of cpn60-based real-time quantitative PCR assays for the detection of 14 Campylobacter species and application to screening of canine fecal samples.

    Science.gov (United States)

    Chaban, Bonnie; Musil, Kristyna M; Himsworth, Chelsea G; Hill, Janet E

    2009-05-01

    Campylobacter species are important organisms in both human and animal health. The identification of Campylobacter currently requires the growth of organisms from complex samples and biochemical identification. In many cases, the condition of the sample being tested and/or the fastidious nature of many Campylobacter species has limited the detection of campylobacters in a laboratory setting. To address this, we have designed a set of real-time quantitative PCR (qPCR) assays to detect and quantify 14 Campylobacter species, C. coli, C. concisus, C. curvus, C. fetus, C. gracilis, C. helveticus, C. hyointestinalis, C. jejuni, C. lari, C. mucosalis, C. rectus, C. showae, C. sputorum, and C. upsaliensis, directly from DNA extracted from feces. By use of a region of the cpn60 (also known as hsp60 or groEL) gene, which encodes the universally conserved 60-kDa chaperonin, species-specific assays were designed and validated. These assays were then employed to determine the prevalence of Campylobacter species in fecal samples from dogs. Fecal samples were found to contain detectable and quantifiable levels of C. fetus, C. gracilis, C. helveticus, C. jejuni, C. showae, and C. upsaliensis, with the majority of samples containing multiple Campylobacter species. This study represents the first report of C. fetus, C. gracilis, C. mucosalis, and C. showae detection in dogs and implicates dogs as a reservoir for these species. The qPCR assays described offer investigators a new tool to study many Campylobacter species in a culture-independent manner.

  19. PCR assay for the detection of Campylobacter in marinated and non-marinated poultry products.

    Science.gov (United States)

    Katzav, Marianne; Isohanni, Pauliina; Lund, Marianne; Hakkinen, Marjaana; Lyhs, Ulrike

    2008-10-01

    During a period of 9 months, 194 marinated and non-marinated poultry products were collected from retail shops in a defined area in Western Finland and tested for Campylobacter spp. using a conventional enrichment culture and Polymerase Chain Reaction (PCR) method. For marinated poultry products, the study involved modification of a commercial DNA isolation method. Using either a conventional culture or PCR method, a total of 25 (12.9%) of all investigated samples were Campylobacter positive. In marinated poultry products, Campylobacter was detected at a prevalence of 21.1% and 9.5% in turkey and chicken products, respectively. In August, there was a peak with 28.9% positive Campylobacter samples. Campylobacter inoculation tests were carried out to test the detection limit of both methods. The PCR method used is faster than microbiological analyses. However, enrichment of the samples is necessary due to the low occurrence of Campylobacter spp. in retail Finnish poultry products.

  20. Detection of virulence, antibiotic resistance and toxin (VAT) genes in Campylobacter species using newly developed multiplex PCR assays.

    Science.gov (United States)

    Laprade, Natacha; Cloutier, Michel; Lapen, David R; Topp, Edward; Wilkes, Graham; Villemur, Richard; Khan, Izhar U H

    2016-05-01

    Campylobacter species are one of the leading causes of bacterial gastroenteritis in humans worldwide. This twofold study was sought to: i) develop and optimize four single-tube multiplex PCR (mPCR) assays for the detection of six virulence (ciaB, dnaJ, flaA, flaB, pldA and racR), three toxin (cdtA, cdtB and cdtC) and one antibiotic resistance tet(O) genes in thermophilic Campylobacter spp. and ii) apply and evaluate the developed mPCR assays by testing 470 previously identified C. jejuni, C. coli and C. lari isolates from agricultural water. In each mPCR assay, a combination of two or three sets of primer pairs for virulence, antibiotic resistance and toxin (VAT) genes was used and optimized. Assay 1 was developed for the detection of dnaJ, racR and cdtC genes with expected amplification sizes of 720, 584 and 182bp. Assay 2 generated PCR amplicons for tet(O) and cdtA genes of 559 and 370bp. Assay 3 amplified cdtB ciaB, and pldA genes with PCR amplicon sizes of 620, 527 and 385bp. Assay 4 was optimized for flaA and flaB genes that generated PCR amplicons of 855 and 260bp. The primer pairs and optimized PCR protocols did not show interference and/or cross-amplification with each other and generated the expected size of amplification products for each target VAT gene for the C. jejuni ATCC 33291 reference strain. Overall, all ten target VAT genes were detected at a variable frequency in tested isolates of thermophilic Campylobacter spp. where cdtC, flaB, ciaB, cdtB, cdtA and pldA were commonly detected compared to the flaA, racR, dnaJ and tet(O) genes which were detected with less frequency. The developed mPCR assays are simple, rapid, reliable and sensitive tools for simultaneously assessing potential pathogenicity and antibiotic resistance profiling in thermophilic Campylobacter spp. The mPCR assays will be useful in diagnostic and analytical settings for routine screening of VAT characteristics of Campylobacter spp. as well as being applicable in epidemiological

  1. 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.

  2. 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.

  3. Large-Scale Survey of Campylobacter Species in Human Gastroenteritis by PCR and PCR–Enzyme-Linked Immunosorbent Assay

    Science.gov (United States)

    Lawson, A. J.; Logan, J. M. J.; O'neill, G. L.; Desai, M.; Stanley, J.

    1999-01-01

    A PCR-based study of the incidence of enteropathogenic campylobacter infection in humans was done on the basis of a detection and identification algorithm consisting of screening PCRs and species identification by PCR-enzyme-linked immunosorbent assay. This was applied to DNA extracted from 3,738 fecal samples from patients with sporadic cases of acute gastroenteritis, submitted by seven regional Public Health Laboratories in England and Wales over a 2-year period. The sending laboratories had cultured “Campylobacter spp.” from 464 samples. The PCR methodologies detected 492 Campylobacter-positive samples, and the combination of culture and PCR yielded 543 Campylobacter-positive samples. There was identity (overlap) for 413 samples, but 79 PCR-positive samples were culture negative, and 51 culture-positive samples were PCR negative. While there was no statistically significant difference between PCR and culture in detection of C. jejuni-C. coli (PCR, 478 samples; culture, 461 samples), PCR provided unique data about mixed infections and non-C. jejuni and non- C. coli campylobacters. Mixed infections with C. jejuni and C. coli were found in 19 samples, and mixed infection with C. jejuni and C. upsaliensis was found in one sample; this was not apparent from culture. Eleven cases of gastroenteritis were attributed to C. upsaliensis by PCR, three cases were attributed to C. hyointestinalis, and one case was attributed to C. lari. This represents the highest incidence of C. hyointestinalis yet reported from human gastroenteritis, while the low incidence of C. lari suggests that it is less important in this context. PMID:10565897

  4. Evaluation of Various Campylobacter-Specific Quantitative PCR (qPCR) Assays for Detection and Enumeration of Campylobacteraceae in Irrigation Water and Wastewater via a Miniaturized Most-Probable-Number-qPCR Assay.

    Science.gov (United States)

    Banting, Graham S; Braithwaite, Shannon; Scott, Candis; Kim, Jinyong; Jeon, Byeonghwa; Ashbolt, Nicholas; Ruecker, Norma; Tymensen, Lisa; Charest, Jollin; Pintar, Katarina; Checkley, Sylvia; Neumann, Norman F

    2016-08-01

    Campylobacter spp. are the leading cause of bacterial gastroenteritis worldwide, and water is increasingly seen as a risk factor in transmission. Here we describe a most-probable-number (MPN)-quantitative PCR (qPCR) assay in which water samples are centrifuged and aliquoted into microtiter plates and the bacteria are enumerated by qPCR. We observed that commonly used Campylobacter molecular assays produced vastly different detection rates. In irrigation water samples, detection rates varied depending upon the PCR assay and culture method used, as follows: 0% by the de Boer Lv1-16S qPCR assay, 2.5% by the Van Dyke 16S and Jensen glyA qPCR assays, and 75% by the Linton 16S endpoint PCR when cultured at 37°C. Primer/probe specificity was the major confounder, with Arcobacter spp. routinely yielding false-positive results. The primers and PCR conditions described by Van Dyke et al. (M. I. Van Dyke, V. K. Morton, N. L. McLellan, and P. M. Huck, J Appl Microbiol 109:1053-1066, 2010, http://dx.doi.org/10.1111/j.1365-2672.2010.04730.x) proved to be the most sensitive and specific for Campylobacter detection in water. Campylobacter occurrence in irrigation water was found to be very low (Campylobacter-specific qPCR was used, with the most commonly detected species being C. jejuni, C. coli, and C. lari Campylobacters in raw sewage were present at ∼10(2)/100 ml, with incubation at 42°C required for reducing microbial growth competition from arcobacters. Overall, when Campylobacter prevalence and/or concentration in water is reported using molecular methods, considerable validation is recommended when adapting methods largely developed for clinical applications. Furthermore, combining MPN methods with molecular biology-based detection algorithms allows for the detection and quantification of Campylobacter spp. in environmental samples and is potentially suited to quantitative microbial risk assessment for improved public health disease prevention related to food and water

  5. Evaluation of Various Campylobacter-Specific Quantitative PCR (qPCR) Assays for Detection and Enumeration of Campylobacteraceae in Irrigation Water and Wastewater via a Miniaturized Most-Probable-Number–qPCR Assay

    Science.gov (United States)

    Banting, Graham S.; Braithwaite, Shannon; Scott, Candis; Kim, Jinyong; Jeon, Byeonghwa; Ashbolt, Nicholas; Ruecker, Norma; Tymensen, Lisa; Charest, Jollin; Pintar, Katarina; Checkley, Sylvia

    2016-01-01

    ABSTRACT Campylobacter spp. are the leading cause of bacterial gastroenteritis worldwide, and water is increasingly seen as a risk factor in transmission. Here we describe a most-probable-number (MPN)–quantitative PCR (qPCR) assay in which water samples are centrifuged and aliquoted into microtiter plates and the bacteria are enumerated by qPCR. We observed that commonly used Campylobacter molecular assays produced vastly different detection rates. In irrigation water samples, detection rates varied depending upon the PCR assay and culture method used, as follows: 0% by the de Boer Lv1-16S qPCR assay, 2.5% by the Van Dyke 16S and Jensen glyA qPCR assays, and 75% by the Linton 16S endpoint PCR when cultured at 37°C. Primer/probe specificity was the major confounder, with Arcobacter spp. routinely yielding false-positive results. The primers and PCR conditions described by Van Dyke et al. (M. I. Van Dyke, V. K. Morton, N. L. McLellan, and P. M. Huck, J Appl Microbiol 109:1053–1066, 2010, http://dx.doi.org/10.1111/j.1365-2672.2010.04730.x) proved to be the most sensitive and specific for Campylobacter detection in water. Campylobacter occurrence in irrigation water was found to be very low (Campylobacter-specific qPCR was used, with the most commonly detected species being C. jejuni, C. coli, and C. lari. Campylobacters in raw sewage were present at ∼102/100 ml, with incubation at 42°C required for reducing microbial growth competition from arcobacters. Overall, when Campylobacter prevalence and/or concentration in water is reported using molecular methods, considerable validation is recommended when adapting methods largely developed for clinical applications. Furthermore, combining MPN methods with molecular biology-based detection algorithms allows for the detection and quantification of Campylobacter spp. in environmental samples and is potentially suited to quantitative microbial risk assessment for improved public health disease prevention related to

  6. PCR assay for the detection of Campylobacter in marinated and non-marinated poultry products

    DEFF Research Database (Denmark)

    Katzav, Marianne; Isohanni, Pauliina; Lund, Marianne

    2008-01-01

    During a period of 9 months, 194 marinated and non-marinated poultry products were collected from retail shops in a defined area in Western Finland and tested for Campylobacter spp. using a conventional enrichment culture and Polymerase Chain Reaction (PCR) method. For marinated poultry products......, the study involved modification of a commercial DNA isolation method. Using either a conventional culture or PCR method, a total of 25 (12.9%) of all investigated samples were Campylobacter positive. In marinated poultry products, Campylobacter was detected at a prevalence of 21.1% and 9.5% in turkey...

  7. Survival rates of thermotolerant Campylobacter species in a transport and enrichment medium under different environmental conditions Taxas de sobrevida de espécies termotolerantes de Campylobacter mantidas em um meio de transporte sob diferentes condições ambientais

    Directory of Open Access Journals (Sweden)

    A. Tresierra-Ayala

    2006-08-01

    Full Text Available Determinou-se a sobrevida de Campylobacter jejuni subsp. jejuni e C. coli no meio de transporte e enriquecimento TEC, mantido sob diferentes condições de temperatura e concentração de oxigênio. A sobrevida da maioria das amostras foi superior a cinco dias, obtendo-se os períodos de sobrevida mais prolongados (sete a 15 dias, quando o meio foi incubado em microaerofilia à temperatura ambiente, condições nas quais o tempo de sobrevida de C. coli foi superior ao de C. jejuni subsp. jejuni.

  8. PCR assay for the detection of Campylobacter in marinated and non-marinated poultry products

    OpenAIRE

    Katzav, Marianne; Isohanni, Pauliina; Lund, Marianne; Hakkinen, Marjaana; Lyhs, Ulrike

    2008-01-01

    During a period of 9 months, 194 marinated and non-marinated poultry products were collected from retail shops in a defined area in Western Finland and tested for Campylobacter spp. using a conventional enrichment culture and Polymerase Chain Reaction (PCR) method. For marinated poultry products, the study involved modification of a commercial DNA isolation method. Using either a conventional culture or PCR method, a total of 25 (12.9%) of all investigated samples were Campylobacter positive....

  9. A Cytolethal Distending Toxin Gene-Based Multiplex PCR Assay for Campylobacter jejuni, C. fetus, C. coli, C. upsaliensis, C. hyointestinalis, and C. lari.

    Science.gov (United States)

    Kamei, Kazumasa; Kawabata, Hiroki; Asakura, Masahiro; Samosornsuk, Worada; Hinenoya, Atsushi; Nakagawa, Shinsaku; Yamasaki, Shinji

    2016-05-20

    In this study, we devised a multiplex PCR assay based on the gene of cytolethal distending toxin (cdt) B subunit to simultaneously detect and discriminate Campylobacter jejuni, C. fetus, C. coli, C. upsaliensis, C. hyointestinalis, and C. lari. Species-specific PCR products were successfully obtained from all 38 C. jejuni, 12 C. fetus, 39 C. coli, 22 C. upsaliensis, 24 C. hyointestinalis, and 7 C. lari strains tested. On the other hand, no specific PCR products were obtained from other campylobacters and bacterial species tested (41 strains in total). The proposed multiplex PCR assay is a valuable tool for detection and descrimination of 6 major Campylobacter species, that are associated with gastrointestinal diseases in humans.

  10. Application of a multiplex PCR assay for Campylobacter fetus detection and subspecies differentiation in uncultured samples of aborted bovine fetuses.

    Science.gov (United States)

    Iraola, Gregorio; Hernández, Martín; Calleros, Lucía; Paolicchi, Fernando; Silveyra, Silvia; Velilla, Alejandra; Carretto, Luis; Rodríguez, Eliana; Pérez, Ruben

    2012-12-01

    Campylobacter (C.) fetus (epsilonproteobacteria) is an important veterinary pathogen. This species is currently divided into C. fetus subspecies (subsp.) fetus (Cff) and C. fetus subsp. venerealis (Cfv). Cfv is the causative agent of bovine genital Campylobacteriosis, an infectious disease that leads to severe reproductive problems in cattle worldwide. Cff is a more general pathogen that causes reproductive problems mainly in sheep although cattle can also be affected. Here we describe a multiplex PCR method to detect C. fetus and differentiate between subspecies in a single step. The assay was standardized using cultured strains and successfully used to analyze the abomasal liquid of aborted bovine fetuses without any pre-enrichment step. Results of our assay were completely consistent with those of traditional bacteriological diagnostic methods. Furthermore, the multiplex PCR technique we developed may be easily adopted by any molecular diagnostic laboratory as a complementary tool for detecting C. fetus subspecies and obtaining epidemiological information about abortion events in cattle.

  11. Bacterial cell-surface displaying of thermo-tolerant glutamate dehydrogenase and its application in L-glutamate assay.

    Science.gov (United States)

    Song, Jianxia; Liang, Bo; Han, Dongfei; Tang, Xiangjiang; Lang, Qiaolin; Feng, Ruirui; Han, Lihui; Liu, Aihua

    2015-03-01

    In this paper, glutamate dehydrogenase (Gldh) is reported to efficiently display on Escherichia coli cell surface by using N-terminal region of ice the nucleation protein as an anchoring motif. The presence of Gldh was confirmed by SDS-PAGE and enzyme activity assay. Gldh was detected mainly in the outer membrane fraction, suggesting that the Gldh was displayed on the bacterial cell surface. The optimal temperature and pH for the bacteria cell-surface displayed Gldh (bacteria-Gldh) were 70°C and 9.0, respectively. Additionally, the fusion protein retained almost 100% of its initial enzymatic activity after 1 month incubation at 4°C. Transition metal ions could inhibit the enzyme activity to different extents, while common anions had little adverse effect on enzyme activity. Importantly, the displayed Gldh is most specific to l-glutamate reported so far. The bacterial Gldh was enabled to catalyze oxidization of l-glutamate with NADP(+) as cofactor, and the resultant NADPH can be detected spectrometrically at 340nm. The bacterial-Gldh based l-glutamate assay was established, where the absorbance at 340nm increased linearly with the increasing l-glutamate concentration within the range of 10-400μM. Further, the proposed approach was successfully applied to measure l-glutamate in real samples. Copyright © 2014 Elsevier Inc. All rights reserved.

  12. Development of a novel triplex PCR assay for the detection and differentiation of thermophilic species of Campylobacter using 16S-23S rDNA internal transcribed spacer (ITS) region.

    Science.gov (United States)

    Khan, I U H; Edge, T A

    2007-12-01

    Campylobacter species are significantly implicated in human gastrointestinal infections. Of 20 species of Campylobacter, C. jejuni, C. coli and C. lari have been considered as the most important causative agents of human infections. In order to better understand the occurrence and epidemiology of these thermophilic Campylobacter species, an improved and rapid detection method is warranted. A novel triplex polymerase chain reaction (PCR) assay was developed based on the variable 16S-23S rDNA internal transcribed spacer (ITS) region to identify and discriminate between these species in water samples. Campylobacter species-specific primers for C. jejuni, C. coli and C. lari derived from highly variable sequences in the ITS region were used. Specificity of the newly designed primers and PCR conditions were verified using other species of Campylobacter as well as 31 different negative control species. The assay was further validated with 97 Campylobacter cultures from water samples. The assay was found to be simple, easy to perform, and had a high sensitivity, specificity and reproducibility. It enabled simultaneous detection and differentiation of multiple Campylobacter species in water samples. Use of the newly developed PCR assay, coupled with a previously developed rapid DNA template preparation step, will enable improved detection capabilities for Campylobacter species in environmental matrices.

  13. Evaluation of PCR assays for the detection of Campylobacter fetus in bovine preputial scrapings and the identification of subspecies in South African field isolates.

    Science.gov (United States)

    Schmidt, T; Venter, E H; Picard, J A

    2010-06-01

    As a result of the high lability and slow growth of Campylobacter fetus subspecies, the laboratory diagnosis of bovine genital campylobacteriosis has always been difficult. This is especially true under South African conditions, where farms are far apart, laboratories are only present in major centres and there are high ambient temperatures. In order to overcome the shortcomings associated with traditional diagnostic methods, the implementation of a molecular assay was sought. This work describes how a previously published PCR assay (MG3F/ MG4R primers) was adapted, optimised and applied in the diagnostic laboratory to test preputial samples directly for the presence of Campylobacter fetus. Field evaluation of the assay revealed an analytical sensitivity and specificity of 85.7% and 99%, respectively. Subsequent genotyping and phenotyping of a diverse collection of South African field isolates revealed that South Africa has an unexpected and previously unreported high incidence of Campylobacter fetus subsp. venerealis biovar intermedius strains. These strains were not identified correctly by the subspecies-specific primer set evaluated. Until such time that cost- effective genotyping methods are available to diagnostic laboratories in South Africa, and other countries with these atypical Campylobacter fetus subsp. venerealis strains, the need for bacterial culture will persist. Identification to subspecies level of isolates at present remains dependent upon a single phenotypic criterion, namely tolerance to 1% glycine.

  14. Evaluation of PCR assays for the detection of Campylobacter fetus in bovine preputial scrapings and the identification of subspecies in South African field isolates

    Directory of Open Access Journals (Sweden)

    T. Schmidt

    2010-05-01

    Full Text Available As a result of the high lability and slow growth of Campylobacter fetus subspecies, the laboratory diagnosis of bovine genital campylobacteriosis has always been difficult. This is especially true under South African conditions, where farms are far apart, laboratories are only present in major centres and there are high ambient temperatures. In order to overcome the shortcomings associated with traditional diagnostic methods, the implementation of a molecular assay was sought. This work describes how a previously published PCR assay (MG3F / MG4R primers was adapted, optimised and applied in the diagnostic laboratory to test preputial samples directly for the presence of Campylobacter fetus. Field evaluation of the assay revealed an analytical sensitivity and specificity of 85.7 % and 99 %, respectively. Subsequent genotyping and phenotyping of a diverse collection of South African field isolates revealed that South Africa has an unexpected and previously unreported high incidence of Campylobacter fetus subsp. venerealis biovar intermedius strains. These strains were not identified correctly by the subspecies-specific primer set evaluated. Until such time that cost-effective genotyping methods are available to diagnostic laboratories in South Africa, and other countries with these atypical Campylobacter fetus subsp. venerealis strains, the need for bacterial culture will persist. Identification to subspecies level of isolates at present remains dependent upon a single phenotypic criterion, namely tolerance to 1 % glycine.

  15. Development and evaluation of a PCR assay for rapid detection of azithromycin resistant Campylobacter isolated from diarrhoeal patients in Kolkata, India.

    Science.gov (United States)

    Mukherjee, Piyali; Dutta, Shanta; Mukhopadhyay, Asish K

    2017-01-01

    Campylobacter is a well-known bacterial pathogen for triggering acute gastroenteritis in humans both in developed and developing countries. This organism is highly resistant to fluoroquinolones. Macrolides are very much useful for the treatment of campylobacteriosis when clinical therapy is necessary. However, increasing resistance to azithromycin, a potent macrolide has been reported in Campylobacter in recent years. Macrolide resistance in Campylobacter is found mainly due to point mutation in V region of 23S rRNA. We have developed a PCR based assay, which can detect the azithromycin resistant and sensitive Campylobacter strains utilizing mutation responsible for the phenotype. This PCR was validated using 359 Campylobacter strains isolated from diarrhoeal patients at Kolkata, India. Antimicrobial resistance through disk diffusion method was also performed on these strains as a gold standard. Studies through sequencing analysis further confirmed the PCR result. This study describes a simple and rapid method for detection of mutation conferring macrolide resistance with additional feature of identification of sensitive strains.

  16. A cytolethal distending toxin gene-based multiplex PCR assay for detection of Campylobacter spp. in stool specimens and comparison with culture method.

    Science.gov (United States)

    Shiramaru, Sachi; Asakura, Masahiro; Inoue, Haruna; Nagita, Akira; Matsuhisa, Akio; Yamasaki, Shinji

    2012-07-01

    In this study, we evaluated the applicability of cytolethal distending toxin (cdt) gene-based species-specific multiplex PCR for the direct detection and identification of Campylobacter jejuni, C. coli and C. fetus from stool specimens of patients with gastroenteritis in comparison to culture methods. A total of 711 stool specimens were examined for the isolation or detection of campylobacters by using Skirrow's selective agar culture plates, a filtration method and the multiplex PCR assay. Forty-one and 36 C. jejuni strains were isolated by culture and filtration methods, respectively. In addition, 2 and 3 C. coli strains were isolated by Skirrow and the filtration methods, respectively. However, when the multiplex PCR was employed, the cdtB genes of C. jejuni and C. coli were detected in 45 and 4 stool samples, respectively, and 9 C. jejuni PCR-positive samples by multiplex PCR were negative by culture method. Sequence analysis of the PCR products obtained from 8 stool specimens from which campylobacters were not isolated by culture method but the sequences exactly matched with that of the cdtB gene of C. jejuni strain 81-176. None of the remaining stool samples which were culture negative for campylobacters produced any amplicon. Stool samples were defined as Campylobacter-positive if detected by any method. The sensitivity of the multiplex PCR was 83%, which was higher than Skirrow (74%) and filtration method (66%). These data indicate that cdtB gene-based multiplex PCR is a rapid and more sensitive method to identify the most important species of Campylobacter for human diseases. (248).

  17. 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.

  18. Multiplex PCR assay for the detection and quantification of Campylobacter spp., Escherichia coli O157:H7, and Salmonella serotypes in water samples.

    Science.gov (United States)

    Park, Si Hong; Hanning, Irene; Jarquin, Robin; Moore, Philip; Donoghue, Dan J; Donoghue, Ann M; Ricke, Steven C

    2011-03-01

    Three pathogens, Campylobacter, Salmonella, and Shiga-toxin-producing Escherichia coli, are leading causes of bacterial gastroenteritis in the United States and worldwide. Although these three bacteria are typically considered food-borne pathogens, outbreaks have been reported due to contaminated drinking water and irrigation water. The aim of this research was to develop two types of PCR assays that could detect and quantify three pathogens, Campylobacter spp., E. coli O157:H7, and Salmonella spp., in watershed samples. In conventional PCR, three target strains were detected by multiplex PCR (m-PCR) using each specific primer pair simultaneously. Under optimized m-PCR conditions, the assay produced a 90-bp product for Campylobacter jejuni, a 150-bp product for E. coli O157:H7, and a 262-bp product for Salmonella Typhimurium, and the limitation of detection was approximately 700 copies for all three bacteria. In addition, real-time PCR was performed to quantify the three pathogens using SYBR green fluorescence. The assay was designed so that each target had a different melting temperature [C. jejuni (80.1 °C), E. coli O157:H7 (83.3 °C), and S. Typhimurium (85.9 °C)]. Therefore, this system could quantify and distinguish three pathogens simultaneously in a single reaction. © 2011 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.

  19. Determination of ciprofloxacin and nalidixic acid resistance in Campylobacter jejuni with a fluorogenic polymerase chain reaction assay.

    Science.gov (United States)

    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).

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

    Directory of Open Access Journals (Sweden)

    Harkanwaldeep Singh

    2011-03-01

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

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

    Science.gov (United States)

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

    2011-01-01

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

  2. 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,

  3. 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.

  4. Evaluation of a cytolethal distending toxin (cdt) gene-based species-specific multiplex PCR assay for the identification of Campylobacter strains isolated from diarrheal patients in Japan.

    Science.gov (United States)

    Kabir, S M Lutful; Kikuchi, Ken; Asakura, Masahiro; Shiramaru, Sachi; Tsuruoka, Naoki; Goto, Aeko; Hinenoya, Atsushi; Yamasaki, Shinji

    2011-01-01

    We have developed a cytolethal distending toxin (cdt) gene-based species-specific multiplex PCR assay for the detection and identification of Campylobacter jejuni, C. coli, and C. fetus. The applicability of this assay was evaluated with 325 Campylobacter strains isolated from diarrheal patients in Japan and the results were compared with those obtained by other genetic methods, including hipO gene detection and 16S rRNA gene sequencing. Of the 325 strains analyzed, 314 and 11 were identified as C. jejuni and C. coli, respectively, by combination of hipO gene detection and 16S rRNA gene sequencing. When the multiplex PCR assay was employed, 309, 310, and 314 strains were identified as C. jejuni on the basis of cdtA, cdtB, and cdtC gene-specific primers, respectively. Similarly, 11, 11, and 10 strains were identified as C. coli on the basis of cdtA, cdtB, and cdtC gene-specific primers, respectively. Sequence analysis of the cdt gene region of 6 strains (5 C. jejuni and 1 C. coli) which did not yield specific PCR products in any of the cdt gene-based multiplex PCR assays revealed deletions or mutations of the cdt genes. Pulsed-field gel electrophoresis indicated that C. jejuni and C. coli strains were genetically diverse. Taken together, these findings suggest that the cdtC gene-based multiplex PCR seems to be a particularly simple and rapid method for differentiating between species of Campylobacter strains, such as C. jejuni and C. coli. However, combination of these multiplex PCR assays will allow more accurate identification.

  5. 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

  6. 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, ...

  7. Complete genome sequence of the hippuricase-positive Campylobacter avium type strain LMG 24591

    Science.gov (United States)

    Campylobacter avium is a hippurate-positive, thermotolerant campylobacter that has been isolated from poultry. Here we present the genome sequences of two C. avium strains isolated from broiler chickens: strains LMG 24591T (complete genome) and LMG 24592 (draft genome). The C. avium type strain geno...

  8. Comparative genomics of all three Campylobacter sputorum biovars and a novel cattle-associated C. sputorum clade

    Science.gov (United States)

    Campylobacter sputorum is a non-thermotolerant campylobacter that is primarily isolated from food animals such as cattle and sheep. C. sputorum is also infrequently associated with human illness. Based on catalase and urease activity, three biovars are currently recognized within C. sputorum: bv. sp...

  9. Detection of pathogenic Campylobacter, E. coli O157:H7 and Salmonella spp. in wastewater by PCR assay.

    Science.gov (United States)

    Bonetta, Si; Pignata, C; Lorenzi, E; De Ceglia, M; Meucci, L; Bonetta, Sa; Gilli, G; Carraro, E

    2016-08-01

    The aim of this study was the evaluation of the occurrence of pathogenic Campylobacter, Escherichia coli O157:H7, E. coli virulence genes and Salmonella spp. in different wastewater treatment plants (WWTPs) using a method based on an enrichment step and PCR. This method was sensitive enough to detect low levels (∼2 CFU100 ml(-1) of raw sewage) of all the investigated pathogens. In the WWTP samples, E. coli O157:H7 DNA and the eae gene were never found, but 33 % of influents and effluents exhibited amplicons corresponding to Shiga-like toxin I. Twenty-five percent of the influent and 8 % of the effluent exhibited the presence of Shiga-like toxin II. Campylobacter jejuni and C. coli DNA were identified in 50 and 25 % of the influents and in 8 and 25 % of the effluents, respectively. Salmonella spp. DNA was present in all the samples. Considering the results obtained, the method tested here offers a reliable and expeditious tool for evaluating the efficiency of the effluent treatment in order to mitigate contamination risk. Influent contamination by Salmonella spp. and Campylobacter spp. provides indirect information about their circulation; moreover, their presence in effluents underlines the role of WWTPs in the contamination of the receiving surface waters, which affects public health directly or indirectly.

  10. Pyrosequencing-based validation of a simple cell-suspension polymerase chain reaction assay for Campylobacter with application of high-processivity polymerase and novel internal amplification controls for rapid and specific detection.

    Science.gov (United States)

    Oakley, Brian B; Line, J Eric; Berrang, Mark E; Johnson, Jessica M; Buhr, R Jeff; Cox, Nelson A; Hiett, Kelli L; Seal, Bruce S

    2012-02-01

    Although Campylobacter is an important food-borne human pathogen, there remains a lack of molecular diagnostic assays that are simple to use, cost-effective, and provide rapid results in research, clinical, or regulatory laboratories. Of the numerous Campylobacter assays that do exist, to our knowledge none has been empirically tested for specificity using high-throughput sequencing. Here we demonstrate the power of next-generation sequencing to determine the specificity of a widely cited Campylobacter-specific polymerase chain reaction (PCR) assay and describe a rapid method for direct cell suspension PCR to quickly and easily screen samples for Campylobacter. We present a specific protocol which eliminates the need for time-consuming and expensive genomic DNA extractions and, using a high-processivity polymerase, demonstrate conclusive screening of samples in 99%) sensitive, and spike-back experiments demonstrated a detection threshold of <10(2) CFU mL(-1). Additionally, we present 2 newly designed broad-range bacterial primer sets targeting the 23S rRNA gene that have wide applicability as internal amplification controls. Empirical testing of putative taxon-specific assays using high-throughput sequencing is an important validation step that is now financially feasible for research, regulatory, or clinical applications. Published by Elsevier Inc.

  11. Development of Multiplex-Mismatch Amplification Mutation-PCR Assay for Simultaneous Detection of Campylobacter jejuni and Mutation in gyrA Gene Related to Fluoroquinolone Resistance.

    Science.gov (United States)

    Cui, Mingquan; Wu, Chenbin; Zhang, Peng; Wu, Congming

    2016-11-01

    Campylobacter jejuni, a foodborne pathogen, is the major cause of enteritis in humans worldwide, however, its increasing resistance to fluoroquinolones reported recently is of a major concern. In the present study, multiplex-mismatch amplification mutation assay-polymerase chain reaction (MMAMA-PCR) was developed for the first time with the aim to quickly identify C. jejuni and to detect the single nucleotide mutation (C-257 to T) frequently observed in gyrA gene, associated with the acquisition of resistance to fluoroquinolones. In this assay, mismatch amplification mutation primers for the detection of gyrA mutation in C. jejuni were coupled with primers for the hip gene encoding for hippuricase and 16S rRNA gene of C. jejuni, respectively, in the multiplex PCR assay. The specificity and accuracy of this method were analyzed by the use of 78 C. jejuni strains with previously confirmed resistance phenotypes and the mutation (C-257 to T) in gyrA gene, as well as 107 clinical isolates of various bacterial species, including 29 C. jejuni isolates. This study indicates that MMAMA-PCR is a promising assay for the rapid identification of C. jejuni with a specific mutation in gyrA gene, responsible for the resistance to fluoroquinolones.

  12. Danish strategies to control Campylobacter in broilers and broiler meat: facts and effects

    DEFF Research Database (Denmark)

    Rosenquist, Hanne; Boysen, Louise; Galliano, C.

    2009-01-01

    Thermotolerant Campylobacter spp. have been the most common bacterial cause of human gastrointestinal disease in Denmark since 1999. In 2003, the Danish voluntary strategy to control Campylobacter was intensified. The focus was on biosecurity, allocation of meat from Campylobacter-negative broilers...... to the production of chilled products, and consumer information campaigns. From 2002 to 2007, the percentage of Campylobacter-positive broiler flocks at slaughter decreased from 43% to 27%. After processing, Campylobacter-positive samples of chilled broiler meat fell from 18% in 2004 to 8% in 2007. Furthermore......, the number of registered human Campylobacter cases decreased by 12%; from 4379 cases in 2002 to 3865 cases in 2007. We believe that the observed decrease in the occurrence of Campylobacter in broilers and broiler meat and the coincidental fall in the number of registered human cases is, in part, a result...

  13. Development of multiplex real-time PCR assay for the detection of Brucella spp., Leptospira spp. and Campylobacter foetus.

    Science.gov (United States)

    Selim, Abdelfattah M; Elhaig, Mahmoud M; Gaede, Wolfgang

    2014-12-29

    Abortion among dairy cattle is one of the major causes of economic losses in the livestock industry. This study describes a 1-step multiplex real-time polymerase chain reaction (PCR) to detect Brucella spp., Leptospira spp. and Campylobacter foetus, these are significant bacteria commonly implicated in bovine abortion. ß-actin was added to the same PCR reaction as an internal control to detect any extraction failure or PCR inhibition. The detection limit of multiplex real-time PCR using purified DNA from cultured organisms was set to 5 fg for Leptospira spp. and C. foetus and to 50 fg for Brucella spp. The multiplex real-time PCR did not produce any non-specific amplification when tested with different strains of the 3 pathogens. This multiplex real-time PCR provides a valuable tool for diagnosis, simultaneous and rapid detection for the 3 pathogens causing abortion in bovine.

  14. Development of multiplex real-time PCR assay for the detection of Brucella spp., Leptospira spp. and Campylobacter foetus

    Directory of Open Access Journals (Sweden)

    Abdelfattah M. Selim

    2014-12-01

    Full Text Available Abortion among dairy cattle is one of the major causes of economic losses in the livestock industry. This study describes a 1-step multiplex real-time polymerase chain reaction (PCR to detect Brucella spp., Leptospira spp. and Campylobacter foetus, these are significant bacteria commonly implicated in bovine abortion. ß-actin was added to the same PCR reaction as an internal control to detect any extraction failure or PCR inhibition. The detection limit of multiplex real-time PCR using purified DNA from cultured organisms was set to 5 fg for Leptospira spp. and C. foetus and to 50 fg for Brucella spp. The multiplex real-time PCR did not produce any non-specific amplification when tested with different strains of the 3 pathogens. This multiplex real-time PCR provides a valuable tool for diagnosis, simultaneous and rapid detection for the 3 pathogens causing abortion in bovine.

  15. The complete genome sequence and analysis of the human pathogen Campylobacter lari

    DEFF Research Database (Denmark)

    Miller, WG; Wang, G; Binnewies, Tim Terence

    2008-01-01

    Campylobacter lari is a member of the epsilon subdivision of the Proteobacteria and is part of the thermotolerant Campylobacter group, a clade that includes the human pathogen C. jejuni. Here we present the complete genome sequence of the human clinical isolate, C. lari RM2100. The genome of strain...... RM2100 is approximately 1.53 Mb and includes the 46 kb megaplasmid pCL2100. Also present within the strain RM2100 genome is a 36 kb putative prophage, termed CLIE1, which is similar to CJIE4, a putative prophage present within the C. jejuni RM1221 genome. Nearly all (90%) of the gene content...... in strain RM2100 is similar to genes present in the genomes of other characterized thermotolerant campylobacters. However, several genes involved in amino acid biosynthesis and energy metabolism, identified previously in other Campylobacter genomes, are absent from the C. lari RM2100 genome. Therefore, C...

  16. Enzyme-linked immunosorbent assay for Campylobacter pyloridis: correlation with presence of C. pyloridis in the gastric mucosa.

    Science.gov (United States)

    Goodwin, C S; Blincow, E; Peterson, G; Sanderson, C; Cheng, W; Marshall, B; Warren, J R; McCulloch, R

    1987-03-01

    Antibody to Campylobacter pyloridis was measured by ELISA in the sera of 160 patients from whom gastric biopsy specimens were also obtained. The antigen was an acid-glycine extract of C. pyloridis, and titers ranged from 80 to 22,000 ELISA units (EU). Of 117 patients in whom C. pyloridis was detected microbiologically or histologically, 87 (74%) had a titer greater than or equal to 300 EU, and only one had a titer less than 150 EU. Of 43 patients in whom C. pyloridis was not detected, only two (5%) had a titer greater than 300 EU. Thus, for a titer of 300 EU the ELISA test had a specificity of 97% and a sensitivity of 81%. At 150 EU the specificity was 78%, and the sensitivity was 99%. Histological diagnosis of active chronic gastritis was associated with a high median ELISA titer (485 E), chronic gastritis with a much lower titer (150 EU), and normal histology with a titer of 110 EU. Discriminating use of this serological test could be of assistance to detect C. pyloridis in the gastric mucosa.

  17. Campylobacter Infections

    Science.gov (United States)

    Campylobacter infection is a common foodborne illness. You usually get it from eating contaminated food, especially raw ... You can also get it from drinking contaminated water or raw milk, or handling infected animal feces ( ...

  18. Molecular and Statistical Analysis of Campylobacter spp. and Antimicrobial-Resistant Campylobacter Carriage in Wildlife and Livestock from Ontario Farms.

    Science.gov (United States)

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

    2017-05-01

    The objectives of this study were to (i) compare the carriage of Campylobacter and antimicrobial-resistant Campylobacter among livestock and mammalian wildlife on Ontario farms, and (ii) investigate the potential sharing of Campylobacter subtypes between livestock and wildlife. Using data collected from a cross-sectional study of 25 farms in 2010, we assessed associations, using mixed logistic regression models, between Campylobacter and antimicrobial-resistant Campylobacter carriage and the following explanatory variables: animal species (beef, dairy, swine, raccoon, other), farm type (swine, beef, dairy), type of sample (livestock or wildlife) and Campylobacter species (jejuni, coli, other). Models included a random effect to account for clustering by farm where samples were collected. Samples were subtyped using a Campylobacter-specific 40 gene comparative fingerprinting assay. A total of 92 livestock and 107 wildlife faecal samples were collected, and 72% and 27% tested positive for Campylobacter, respectively. Pooled faecal samples from livestock were significantly more likely to test positive for Campylobacter than wildlife samples. Relative to dairy cattle, pig samples were at significantly increased odds of testing positive for Campylobacter. The odds of isolating Campylobacter jejuni from beef cattle samples were significantly greater compared to dairy cattle and raccoon samples. Fifty unique subtypes of Campylobacter were identified, and only one subtype was found in both wildlife and livestock samples. Livestock Campylobacter isolates were significantly more likely to exhibit antimicrobial resistance (AMR) compared to wildlife Campylobacter isolates. Campylobacter jejuni was more likely to exhibit AMR when compared to C. coli. However, C. jejuni isolates were only resistant to tetracycline, and C.  coli isolates exhibited multidrug resistance patterns. Based on differences in prevalence of Campylobacter spp. and resistant Campylobacter between

  19. [Campylobacter and campylobacteriosis: a view from South America].

    Science.gov (United States)

    Fernández, Heriberto

    2011-03-01

    The thermotolerant species of Campylobacter have become very important in public health, especially as agents of infectious diarrhea in human beings. In this brief revision we present part of the available information generated in South America about epidemiological, clinical and bacteriological aspects of campylobacteriosis and we identify some differences between the observed and documented campylobacteriosis in South America compared to those described in industrialized countries.

  20. Campylobacter Infections

    Science.gov (United States)

    ... Head Neck & Nervous System Heart Infections Learning Disabilities Obesity ... Body Campylobacter are a type of bacteria that produce infections in the GI tract. They are a major bacterial cause of diarrheal sickness among children in the United States. You may hear ...

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

    Science.gov (United States)

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

    2016-10-01

    Infections caused by thermotolerant Campylobacter spp. and Salmonella spp. are the leading causes of human gastroenteritis worldwide. Wild birds can act as reservoirs of both pathogens. A survey was carried out to determine the prevalence, genetic diversity and antimicrobial resistance of thermotolerant Campylobacter and Salmonella in waterfowl used as decoys and wild raptors in Andalusia (Southern Spain). The overall prevalence detected for Campylobacter was 5.9% (18/306; CI95%: 3.25-8.52) in decoys and 2.3% (9/387; CI95%: 0.82-3.83) in wild raptors. Isolates were identified as C. jejuni, C. coli and C. lari in both bird groups. Salmonella was isolated in 3.3% (10/306; CI95%: 2.3-4.3) and 4.6% (18/394; CI95%: 3.5-5.6) of the decoys and raptors, respectively. Salmonella Enteritidis and Typhimurium were the most frequently identified serovars, although Salmonella serovars Anatum, Bredeney, London and Mikawasima were also isolated. Pulsed-field gel electrophoresis analysis of isolates showed higher genetic diversity within Campylobacter species compared to Salmonella serovars. Campylobacter isolates showed resistance to gentamicin, ciprofloxacin and tetracycline, while resistance to erythromycin and tetracycline was found in Salmonella isolates. The results indicate that both decoys and raptors can act as natural carriers of Campylobacter and Salmonella in Spain, which may have important implications for public and animal health. Copyright © 2016 Elsevier Ltd. All rights reserved.

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

    Science.gov (United States)

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

    2015-05-01

    Diarrhea due to enteric bacterial pathogens causes significant morbidity and mortality in the United States and worldwide. However, bacterial pathogens may be infrequently identified. Currently, culture and enzyme immunoassays (EIAs) are the primary methods used by clinical laboratories to detect enteric bacterial pathogens. We conducted a multicenter evaluation of the BD Max enteric bacterial panel (EBP) PCR assay in comparison to culture for the detection of Salmonella spp., Shigella spp., Campylobacter jejuni, and Campylobacter coli and an EIA for Shiga toxins 1 and 2. A total of 4,242 preserved or unpreserved stool specimens, including 3,457 specimens collected prospectively and 785 frozen, retrospective samples, were evaluated. Compared to culture or EIA, the positive percent agreement (PPA) and negative percent agreement (NPA) values for the BD Max EBP assay for all specimens combined were as follows: 97.1% and 99.2% for Salmonella spp., 99.1% and 99.7% for Shigella spp., 97.2% and 98.4% for C. jejuni and C. coli, and 97.4% and 99.3% for Shiga toxins, respectively. Discrepant results for prospective samples were resolved with alternate PCR assays and bidirectional sequencing of amplicons. Following discrepant analysis, PPA and NPA values were as follows: 97.3% and 99.8% for Salmonella spp., 99.2% and 100% for Shigella spp., 97.5% and 99.0% for C. jejuni and C. coli, and 100% and 99.7% for Shiga toxins, respectively. No differences in detection were observed for samples preserved in Cary-Blair medium and unpreserved samples. In this large, multicenter study, the BD Max EBP assay showed superior sensitivity compared to conventional methods and excellent specificity for the detection of enteric bacterial pathogens in stool specimens. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  3. Quantification of Campylobacter spp. in chicken rinse samples by using flotation prior to real-time PCR.

    Science.gov (United States)

    Wolffs, Petra; Norling, Börje; Hoorfar, Jeffrey; Griffiths, Mansel; Rådström, Peter

    2005-10-01

    Real-time PCR is fast, sensitive, specific, and can deliver quantitative data; however, two disadvantages are that this technology is sensitive to inhibition by food and that it does not distinguish between DNA originating from viable, viable nonculturable (VNC), and dead cells. For this reason, real-time PCR has been combined with a novel discontinuous buoyant density gradient method, called flotation, in order to allow detection of only viable and VNC cells of thermotolerant campylobacters in chicken rinse samples. Studying the buoyant densities of different Campylobacter spp. showed that densities changed at different time points during growth; however, all varied between 1.065 and 1.109 g/ml. These data were then used to develop a flotation assay. Results showed that after flotation and real-time PCR, cell concentrations as low as 8.6 x 10(2) CFU/ml could be detected without culture enrichment and amounts as low as 2.6 x 10(3) CFU/ml could be quantified. Furthermore, subjecting viable cells and dead cells to flotation showed that viable cells were recovered after flotation treatment but that dead cells and/or their DNA was not detected. Also, when samples containing VNC cells mixed with dead cells were treated with flotation after storage at 4 or 20 degrees C for 21 days, a similar percentage resembling the VNC cell fraction was detected using real-time PCR and 5-cyano-2,3-ditolyl tetrazolium chloride-4',6'-diamidino-2-phenylindole staining (20% +/- 9% and 23% +/- 4%, respectively, at 4 degrees C; 11% +/- 4% and 10% +/- 2%, respectively, at 20 degrees C). This indicated that viable and VNC Campylobacter cells could be positively selected and quantified using the flotation method.

  4. Prevalence of Campylobacter spp. in poultry meat and meat products imported in Republic of Macedonia

    Directory of Open Access Journals (Sweden)

    Kostova Sandra

    2008-11-01

    Full Text Available Campylobacter spp. is leading bacterial cause of diarrhea in human population in all parts of the world. In most of the cases infection with Campylobacter spp. in humans originate from contaminated poultry meat and poultry meat products. This study was designed to estimate prevalence of Campylobacter spp. in meat and meat products imported in Republic of Macedonia. During the period of 8 months (January-August 2008 we tested 56 samples of meat and meat products (poultry meat, MDM, pork meat, beef meat and smoked beef. Samples were submitted to analysis for detection of thermo-tolerant Campylobacter spp. according to ISO 10272:1995. We determined among the analyzed samples highest prevalence of Campylobacter spp. in MDM with 84% positive samples, poultry meat with 81,8%, pork meat with 10%. We didn.t detect any positive samples in beef meat and smoked beef. Overall prevalence of Campylobacter spp. in all tested samples was 55,36%. This study shows that the high prevalence of Campylobacter spp. in tested samples and in correlation with severe symptoms in humans are reasons good enough for the producing and processing poultry meat industry and food business operators so they should take in consideration Campylobacter spp. in their risk assessment and preparation of HACCP plan.

  5. Campylobacter fetus

    Science.gov (United States)

    Ishihara, Ayaka; Hashimoto, Etaro; Ishioka, Haruhiko; Kobayashi, Hiroyuki; Gomi, Harumi

    2018-01-01

    Meningitis caused by the zoonotic pathogen Campylobacter fetus in immunocompetent adults is rare. We report a 48-year-old Japanese woman with no underlying disease who was found to have meningitis caused by C. fetus . Both C. fetus subsp. fetus and C. fetus subsp. venerealis were isolated from the cerebrospinal fluid culture. The mode of infection in our patient was considered to be associated with the consumption of raw beef and raw cattle liver on a regular basis. Public awareness and education to avoid the consumption of raw or undercooked meat might help prevent C. fetus meningitis.

  6. Veriflow Campylobacter. Performance tested method 101201.

    Science.gov (United States)

    Joelsson, Adam C; Brown, Ashley S; Puri, Amrita; Keough, Martin P; Pascal, Benjamin J; Gaudioso, Zara E

    2014-01-01

    Veriflow Campylobacter is a molecular based assay for the presumptive and qualitative detection of the most common occurring foodborne Campylobacter species: C. jejuni and C. coli. The assay utilizes a PCR detection method coupled with a rapid, visual, flow-based assay that develops in 3 min post PCR amplification and requires only 24 h of non-specialized enrichment for maximum sensitivity. The Veriflow Campylobacter system eliminates the need for microaerobic chambers, gel electrophoresis or fluorophore based detection of target amplification, and does not require complex data analysis. This Performance Tested Method validation study demonstrated the ability of the Veriflow method to detect naturally occurring Campylobacterfrom chicken carcass rinsates. In the reference comparison study, Chi-square and probability of detection analyses of two unpaired studies indicated that there was no significant difference between the Veriflow Campylobacter method and the U.S. Department of Agriculture (USDA)/Food Safety and Inspection Service (FSIS) reference method. There was no indication of false positive or false negative detection in the reference comparison study, and all 50 C. jejuni and C. coli strains were detected, while 35 nonspecific organisms were undetected in the exclusivity/ inclusivity study. The study results show that Veriflow Campylobacter is a sensitive, selective and robust assay for the detection of C. jejuni and C. coli in chicken carcass rinsates.

  7. Campylobacter serology test

    Science.gov (United States)

    ... this page: //medlineplus.gov/ency/article/003530.htm Campylobacter serology test To use the sharing features on this page, please enable JavaScript. Campylobacter serology test is a blood test to look ...

  8. 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 ...

  9. A longitudinal study of Campylobacter distribution in a turkey production chain

    DEFF Research Database (Denmark)

    Perko-Mäkelä, P.; Isohanni, P.; Katzav, M.

    2009-01-01

    . The objective of this study was to determine the occurrence of Campylobacter during a complete turkey production cycle which lasts for 1,5 years of time. For detection of Campylobacter, a conventional culture method was compared with a PCR method. Campylobacter isolates from different types of samples have been...... identified to the species level by a multiplex PCR assay. Methods: Samples (N = 456) were regularly collected from one turkey parent flock, the hatchery, six different commercial turkey farms and from 11 different stages at the slaughterhouse. For the detection of Campylobacter, a conventional culture...... and a PCR method were used. Campylobacter isolates (n = 143) were identified to species level by a multiplex PCR assay. Results: No Campylobacter were detected in either the samples from the turkey parent flock or from hatchery samples using the culture method. PCR detected Campylobacter DNA in five faecal...

  10. Virulence and antibiotic resistance genes in Campylobacter spp. in the Czech Republic.

    Science.gov (United States)

    Bardoň, J; Pudová, V; Koláčková, I; Karpíšková, R; Röderová, M; Kolář, M

    2017-01-01

    Thermotolerant species of the genus Campy-lobacter are the important agents causing human foodborne infections throughout the world. The aims of this study were to evaluate the presence of nine putative virulence genes in Campylobacter spp. isolated from patients and from foods (poultry meat, pork liver), to determine the resistance of Campylobacter isolates to eight antibiotic agents and to detect four resistance genes.Matherial and methods: The presence of the virulence genes cdtA, cdtB, cdtC, virB11, ciaB, wlaN, iam, dnaJ and racR was detected by polymerase chain reaction (PCR) in 94 Campylobacter spp. isolates from humans and 123 campylobacters from foods. The phenotypic resistance to selected antimicrobial agents was tested with microdilution method in 82 human isolates and 91 food isolates. The isolates with antibiograms were tested for the presence of blaOXA-61, tet(O), aph-3-1 and cmeB genes by PCR with specific primers. In both human and food C. jejuni isolates the preva-lence of the studied virulence genes, especially dnaJ, racR, ciaB genes and the toxigenic genes cdtA, cdtB, cdtC, was considerably higher than in C. coli isolates. The only exception was the iam gene identified in only C. coli. The tested isolates of both C. jejuni and C. coli were highly resistant to quinolone antibiotics. Additionally, C. coli was also more resistant to erythromycin, streptomycin and, in case of isolates from pork liver, to tetracycline. High prevalence rates of genes encoding antibiotic resistance was noted for the blaOXA-61 and tet(O) genes in both Campylobacter species. The presented study is the first to assess the presence of genes for virulence and resistance to antibiotics in thermotolerant Campylobacter spp. isolated from humans and foods in the Czech Republic. The resistance of Campylobacter isolates to eight antibiotic agents was also assessed. The prevalence of genes responsible for virulence and resistance is rather varied in thermotolerant Campylobacter spp.

  11. Campylobacter y campylobacteriosis: una mirada desde América del Sur Campylobacter and campylobacteriosis: a view from South America

    Directory of Open Access Journals (Sweden)

    Heriberto Fernández

    2011-03-01

    Full Text Available Las especies termotolerantes de Campylobacter han adquirido gran importancia en la salud pública, por ser considerados agentes de diarrea infecciosa para el ser humano. En esta breve revisión se presenta información sobre aspectos epidemiológicos, clínicos y bacteriológicos de campylobacteriosis en América del Sur. Asimismo, se señalan algunas diferencias con relación a su presentación en países industrializados.The thermotolerant species of Campylobacter have become very important in public health, especially as agents of infectious diarrhea in human beings. In this brief revision we present part of the available information generated in South America about epidemiological, clinical and bacteriological aspects of campylobacteriosis and we identify some differences between the observed and documented campylobacteriosis in South America compared to those described in industrialized countries.

  12. 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...

  13. Application of thermotolerant microorganisms for biofertilizer preparation.

    Science.gov (United States)

    Chen, Kuo-Shu; Lin, Yann-Shying; Yang, Shang-Shyng

    2007-12-01

    Intensive agriculture is practised in Taiwan, and compost application is very popular as a means of improving the soil physical properties and supplying plant nutrition. We tested the potential of inoculation with thermotolerant microorganisms to shorten the maturity and improve the quality of biofertilizer prepared by composting. Thermotolerant microorganisms were isolated from compost and reinoculated for the preparation of biofertilizer. The physical, chemical and biological properties of the biofertilizer were determined during composting. The effects of biofertilizer application on the growth and yield of rape were also studied. Among 3823 colonies of thermotolerant microorganisms, Streptomyces thermonitrificans NTU-88, Streptococcus sp. NTU-130 and Aspergillus fumigatus NTU-132 exhibited high growth rates and cellulolytic and proteolytic activities. When a mixture of rice straw and swine manure were inoculated with these isolates and composted for 61 days, substrate temperature increased initially and then decreased gradually during composting. Substrate pH increased from 7.3 to 8.5. Microbial inoculation enhanced the rate of maturity, and increased the content of ash and total and immobilized nitrogen, improved the germination rate of alfalfa seed, and decreased the content of total organic carbon and the carbon/nitrogen ratio. Biofertilizer application increased the growth and yield of rape. Inoculation of thermotolerant and thermophilic microorganisms to agricultural waste for biofertilizer preparation enhances the rate of maturity and improves the quality of the resulting biofertilizer. Inoculation of appropriate microorganisms in biofertilizer preparation might be usefully applied to agricultural situations.

  14. Use of cellulose filters to isolate Campylobacter spp. from naturally contaminated retail broiler meat.

    Science.gov (United States)

    Speegle, Leslie; Miller, Michael E; Backert, Steffen; Oyarzabal, Omar A

    2009-12-01

    Membrane filtration has been used to isolate Campylobacter spp. from feces, although approximately 5 log CFU/g must be present in the sample. Few studies have attempted to use filter membranes for the isolation of Campylobacter from foods. We investigated the minimum number of thermotolerant Campylobacter cells that pass through cellulose filters, the effect of different cell conditions on the rate of passage, and the minimum number of cells that could pass the filters from enriched broiler meat naturally contaminated with Campylobacter spp. Cellulose filters with 0.65-microm pore sizes retained fewer cells and were more effective than filters with 0.45-microm pore sizes. Scanning electron microscopy revealed that 15 min of contact of the filters with agar plates allowed for the passage of most bacteria. The minimum number of bacteria required to pass through the filters was contingent on cell conditions; nonmotile cells were retained more than motile cells (P blood showed that approximately 1.7 log CFU of Campylobacter can be filtered to pure colonies on agar plates. These results demonstrate that the motility of the bacteria influences passage through cellulose filters and that 0.65-mum-pore-size filters on agar plates help obtain pure Campylobacter colonies from enriched food samples.

  15. 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.

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

    Directory of Open Access Journals (Sweden)

    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.

  17. Fluoroquinolone resistance in Campylobacter

    Science.gov (United States)

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

  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. Flying insects and Campylobacter

    DEFF Research Database (Denmark)

    Hald, Birthe; Sommer, Helle Mølgaard; Skovgård, Henrik

    Campylobacter in flies Flies of the Muscidae family forage on all kind of faeces – various fly species have different preferences. M domestica prefer pigs, horses and cattle faeces, animals which are all known to frequently excrete Campylobacter. As a result, the insects pick up pathogenic micro...... organisms, which may collect on their bodies or survive passage through the fly gut. Campylobacter and other pathogens are then easily transferred to other surfaces, for instance peoples food – or to broiler houses where they may be swallowed by chickens or contaminate the environment. On a large material...... of several species of flies collected outside broiler houses, merely ~1% of the flies were found Campylobacter positive. However, the prevalence varied considerably with fly species, time of the year, and availability of Campylobacter sources. Influx of flies to broiler houses As the influx of flies...

  20. Quantitative and qualitative evaluation of Campylobacter spp. contamination of turkey cecal contents and carcasses during and following the slaughtering process.

    Science.gov (United States)

    Bily, Lise; Petton, Julie; Lalande, Françoise; Rouxel, Sandra; Denis, Martine; Chemaly, Marianne; Salvat, Gilles; Fravalo, Philippe

    2010-07-01

    The present study aimed to document quantitatively and qualitatively the contamination by thermotolerant Campylobacter spp. of turkey samples during slaughtering. Four Campylobacter-positive turkey flocks were investigated at the slaughterhouse at three different stages: evisceration (cecal content), after carcass rinses but before chilling (neck skin), and after breast meat cut (meat). In each case, the studied flock was slaughtered first thing in the morning any given day of the week. The efficiency of cleaning and disinfecting operations was examined in the facility prior to processing the studied flock. For each flock, 90 samples were collected from cecal contents, neck skins, and meat pieces and checked quantitatively and qualitatively for Campylobacter. Identification of Campylobacter species was determined by PCR, and genetic patterns were determined by pulsed-field gel electrophoresis. Campylobacter contamination levels of ceca range from 2 to more than 7 Log CFU/g, while those of neck skin range from 0.5 to 3.5 Log CFU/g and those of meat range from 0.1 to 1.9 Log CFU/g. These differences in Campylobacter counts were not associated with a modification of Campylobacter species ratio; however, in the Campylobacter jejuni population, four genetic groups identified from the ceca were not recovered during slaughtering operations and two other genetic groups were only detected after chilling at the cutting stage of the breast meat. The present study suggests that the slaughtering process did not affect Campylobacter species populations; however, it might have influenced the strain population. Finally, the Campylobacter populations found on breast meat were similar to those isolated from the digestive tract of the birds.

  1. Isolation, cloning and molecular characterization of a thermotolerant ...

    African Journals Online (AJOL)

    Isolation, cloning and molecular characterization of a thermotolerant xylanase from Streptomyces sp. THW31. Thayat Sriyapai, Peechapack Somyoonsap, Supatra Areekit, Paisarn Khawsak, Arda Pakpitcharoen, Kosum Chansiri ...

  2. Recent Advances in Screening of Anti-Campylobacter Activity in Probiotics for Use in Poultry

    Science.gov (United States)

    Saint-Cyr, Manuel J.; Guyard-Nicodème, Muriel; Messaoudi, Soumaya; Chemaly, Marianne; Cappelier, Jean-Michel; Dousset, Xavier; Haddad, Nabila

    2016-01-01

    Campylobacteriosis is the most common cause of bacterial gastroenteritis worldwide. Campylobacter species involved in this infection usually include the thermotolerant species Campylobacter jejuni. The major reservoir for C. jejuni leading to human infections is commercial broiler chickens. Poultry flocks are frequently colonized by C. jejuni without any apparent symptoms. Risk assessment analyses have identified the handling and consumption of poultry meat as one of the most important sources of human campylobacteriosis, so elimination of Campylobacter in the poultry reservoir is a crucial step in the control of this foodborne infection. To date, the use of probiotics has demonstrated promising results to reduce Campylobacter colonization. This review provides recent insights into methods used for probiotic screening to reduce the prevalence and colonization of Campylobacter at the farm level. Different eukaryotic epithelial cell lines are employed to screen probiotics with an anti-Campylobacter activity and yield useful information about the inhibition mechanism involved. These in vitro virulence models involve only human intestinal or cervical cell lines whereas the use of avian cell lines could be a preliminary step to investigate mechanisms of C. jejuni colonization in poultry in the presence of probiotics. In addition, in vivo trials to evaluate the effect of probiotics on Campylobacter colonization are conducted, taking into account the complexity introduced by the host, the feed, and the microbiota. However, the heterogeneity of the protocols used and the short time duration of the experiments lead to results that are difficult to compare and draw conclusions at the slaughter-age of broilers. Nevertheless, the combined approach using complementary in vitro and in vivo tools (cell cultures and animal experiments) leads to a better characterization of probiotic strains and could be employed to assess reduced Campylobacter spp. colonization in chickens if some

  3. Detection, identification and quantification of Campylobacter jejuni, coli and lari in food matrices all at once using multiplex qPCR.

    Science.gov (United States)

    Vondrakova, Lucie; Pazlarova, Jarmila; Demnerova, Katerina

    2014-01-01

    Thermotolerant Campylobacter jejuni, coli and lari are recognized as leading food-borne pathogens causing an acute bacterial enteritis worldwide. Due to narrow spectrum of their biochemical activity, it is very complicated to distinguish between individual species. For reliable risk assessment, proper incidence evaluation or swift sample analysis regarding individual species, a demand for simple and rapid method for their distinguishing is reasonable. In this study, we evaluated a reliable and simple approach for their simultaneous detection, species identification and quantification using multiplex qPCR. Species specific primers and hydrolysis probes are directed to hippuricase gene of C. jejuni, serine hydroxymethyltransferase gene of C. coli and peptidase T gene of C. lari. Efficiencies of reactions were 90.85% for C. jejuni, 96.97% for C. coli and 92.89% for C. lari. At 95.00% confidence level and when cut off is set to 38 cycles, limits of detection are in all cases under 10 genome copies per reaction which is very appreciated since it is known that infectious doses are very low. Proposed assay was positively validated on different food matrices (chicken wing rinses, chicken juice and homogenized fried chicken strips). No inhibition of PCR reaction occurred. Assay was evaluated in accordance with MIQE handbook.

  4. Real Time PCR to detect and differentiate Campylobacter fetus subspecies fetus and Campylobacter fetus subspecies venerealis.

    Science.gov (United States)

    McGoldrick, A; Chanter, J; Gale, S; Parr, J; Toszeghy, M; Line, K

    2013-09-01

    Bovine venereal campylobacter infection, caused by Campylobacter fetus venerealis, is of significant economic importance to the livestock industry. Unfortunately, the successful detection and discrimination of C. fetus venerealis from C. fetus fetus continue to be a limitation throughout the world. There are several publications warning of the problem with biotyping methods as well as with recent molecular based assays. In this study, assessed on 1071 isolates, we report on the successful development of two Real Time SYBR® Green PCR assays that will allow for the detection and discrimination of C. fetus fetus and C. fetus venerealis. The sensitivity reported here for the C. fetus (CampF4/R4) and the C. fetus venerealis (CampF7/R7) specific PCR assays are 100% and 98.7% respectively. The specificity for these same PCR assays are 99.6% and 99.8% respectively. © 2013. Published by Elsevier B.V. All rights reserved.

  5. Concentrations of faecal coliforms, Escherichia coli, enterococci and Campylobacter spp. in equine faeces.

    Science.gov (United States)

    Moriarty, E M; Downing, M; Bellamy, J; Gilpin, B J

    2015-03-01

    To determine the concentration of Campylobacter spp. as well as faecal indicator bacteria; faecal coliforms, Escherichia coli and enterococci in the faeces of healthy adult horses in a sample of properties in the Canterbury region of New Zealand. The faeces of healthy adult horses (n=59), including ponies, pleasure horses and Thoroughbreds, were collected from eight properties around Christchurch, New Zealand. The faeces were analysed for concentrations of Campylobacter spp and faecal indicator bacteria; faecal coliforms, Escherichia coli and enterococci. The presence of other animals on the properties sampled as well as the age, feed and health of the horses at the time of sampling was recorded. Enterococci and faecal coliforms were isolated from all samples, and E. coli was isolated from 58/59 samples. Mean concentrations of faecal coliforms and E. coli did not differ between properties, but there was a significant difference in mean concentration of enterococci between properties. Campylobacter spp. were detected in two faecal samples with one isolate being determined by PCR analysis to be a thermotolerant Campylobacter species, the other C. jejuni. This is the first known report quantifying the concentration of Campylobacter spp. present in healthy adult horses in New Zealand. The presence of equine faecal material in water could elevate concentrations of faecal bacteria and therefore needs to be considered as a source of water contamination. The access of horses to waterways and coastal environments may also need to be restricted to prevent transmission of faecal indicator bacteria and potentially zoonotic agents.

  6. Molecular Detection of Campylobacter spp. in California Gull (Larus californicus) Excreta

    Science.gov (United States)

    We examined the prevalence, quantity, and diversity of Campylobacter species in the excreta of 159 California gull samples using PCR and qPCR based detection assays. While Campylobacter prevalence and abundance was relatively high in the gull excreta examined, molecular data ind...

  7. Pleurotus sajor-caju HSP100 complements a thermotolerance ...

    Indian Academy of Sciences (India)

    To examine its function, PsHsp100 was transformed into a temperature-sensitive hsp104 deletion mutant Saccharomyces cerevisiae strain to test the hypothesis that PsHSP100 is an protein that functions in thermotolerance. Overexpression of PsHSP100 complemented the thermotolerance defect of the hsp104 mutant ...

  8. Isolation and characterization of thermotolerant ethanol-fermenting ...

    African Journals Online (AJOL)

    sunny t

    2016-02-10

    Feb 10, 2016 ... ethanol fermentation ability of the isolated strains were compared with those of the K. marxianus strain DMKU 3-. 1042 as a control, which is one of the most thermotolerant and efficient strains isolated in Thailand (Limtong et al.,. 2007). MATERIALS AND METHODS. Isolation of thermotolerant yeast strains.

  9. Campylobacter contamination and the relative risk of illness from organic broiler meat in comparison with conventional broiler meat

    DEFF Research Database (Denmark)

    Rosenquist, Hanne; Boysen, Louise; Krogh, Anne Louise

    2013-01-01

    Danish organic broiler meat, represented by carcasses sampled at the end of processing after chilling, was more frequently contaminated with thermotolerant Campylobacter spp. than conventional broiler carcasses; the yearly mean prevalence being 54.2% (CI: 40.9-67.5) for organic and 19.7% (CI: 14.......8-24.7) for conventional carcasses. Campylobacter jejuni was the most frequently isolated species. The difference in prevalence was obvious in all quarters of the year. Contamination of organic and conventional broiler carcasses was more likely to occur in the warmer summer months, in this case in the third quarter......, as also documented for conventional broiler flocks. When contaminated, the mean concentration of Campylobacter on neck skin samples of organic and conventional carcasses was not significantly different (P=0.428); 2.0±0.65log10cfu/g and 2.1±0.93log10cfu/g, respectively. Assessing the relative risk...

  10. [Genetics of Campylobacter phages].

    Science.gov (United States)

    Hammerl, Jens A; Jäckel, Claudia; Hertwig, Stefan

    2015-01-01

    The application of virulent (lytic) bacteriophages isa promising tool to reduce the number of Campylobacter along the food chain. However, only little is known aboutthe genetics of Campylobacter phages. To date, the nucleotide sequences of nine virulent Campylobacter phages have been published.The analysis of the sequences indicated that at the nucleotide level, phages of the same group (group II or group III) are closely related, but that similarities between the groups only exist at the protein level. Both groups of phages are distantly related to T4-like phages. The genomes of the studied Campylobacter phages contain numerous genes for homing endonucleases and transposases as well as repetitive sequences. These elements could be important for genomic rearrangements.

  11. 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.

  12. A longitudinal study of Campylobacter distribution in a turkey production chain.

    Science.gov (United States)

    Perko-Mäkelä, Päivikki; Isohanni, Pauliina; Katzav, Marianne; Lund, Marianne; Hänninen, Marja-Liisa; Lyhs, Ulrike

    2009-04-07

    Campylobacter is the most common cause of bacterial enteritis worldwide. Handling and eating of contaminated poultry meat has considered as one of the risk factors for human campylobacteriosis. Campylobacter contamination can occur at all stages of a poultry production cycle. The objective of this study was to determine the occurrence of Campylobacter during a complete turkey production cycle which lasts for 1,5 years of time. For detection of Campylobacter, a conventional culture method was compared with a PCR method. Campylobacter isolates from different types of samples have been identified to the species level by a multiplex PCR assay. Samples (N = 456) were regularly collected from one turkey parent flock, the hatchery, six different commercial turkey farms and from 11 different stages at the slaughterhouse. For the detection of Campylobacter, a conventional culture and a PCR method were used. Campylobacter isolates (n = 143) were identified to species level by a multiplex PCR assay. No Campylobacter were detected in either the samples from the turkey parent flock or from hatchery samples using the culture method. PCR detected Campylobacter DNA in five faecal samples and one fluff and eggshell sample. Six flocks out of 12 commercial turkey flocks where found negative at the farm level but only two were negative at the slaughterhouse. During the brooding period Campylobacter might have contact with the birds without spreading of the contamination within the flock. Contamination of working surfaces and equipment during slaughter of a Campylobacter positive turkey flock can persist and lead to possible contamination of negative flocks even after the end of the day's cleaning and desinfection. Reduction of contamination at farm by a high level of biosecurity control and hygiene may be one of the most efficient ways to reduce the amount of contaminated poultry meat in Finland. Due to the low numbers of Campylobacter in the Finnish turkey production chain

  13. A longitudinal study of Campylobacter distribution in a turkey production chain

    Directory of Open Access Journals (Sweden)

    Hänninen Marja-Liisa

    2009-04-01

    Full Text Available Abstract Background Campylobacter is the most common cause of bacterial enteritis worldwide. Handling and eating of contaminated poultry meat has considered as one of the risk factors for human campylobacteriosis.Campylobacter contamination can occur at all stages of a poultry production cycle. The objective of this study was to determine the occurrence of Campylobacter during a complete turkey production cycle which lasts for 1,5 years of time. For detection of Campylobacter, a conventional culture method was compared with a PCR method. Campylobacter isolates from different types of samples have been identified to the species level by a multiplex PCR assay. Methods Samples (N = 456 were regularly collected from one turkey parent flock, the hatchery, six different commercial turkey farms and from 11 different stages at the slaughterhouse. For the detection of Campylobacter, a conventional culture and a PCR method were used. Campylobacter isolates (n = 143 were identified to species level by a multiplex PCR assay. Results No Campylobacter were detected in either the samples from the turkey parent flock or from hatchery samples using the culture method. PCR detected Campylobacter DNA in five faecal samples and one fluff and eggshell sample. Six flocks out of 12 commercial turkey flocks where found negative at the farm level but only two were negative at the slaughterhouse. Conclusion During the brooding period Campylobacter might have contact with the birds without spreading of the contamination within the flock. Contamination of working surfaces and equipment during slaughter of a Campylobacter positive turkey flock can persist and lead to possible contamination of negative flocks even after the end of the day's cleaning and desinfection. Reduction of contamination at farm by a high level of biosecurity control and hygiene may be one of the most efficient ways to reduce the amount of contaminated poultry meat in Finland. Due to the low numbers of

  14. Epidemiology of Campylobacter in poultry

    NARCIS (Netherlands)

    Jacobs - Reitsma, W.

    1994-01-01

    Campylobacter , causing human infections with severe symptoms of diarrhoea, is mainly transmitted by food, especially poultry meat products.

    Several studies on Campylobacter colonization in breeders, laying hens, and broilers were carried

  15. Influence of physiological environment on the expression of thermotolerance in proliferating (P) and quiescent (Q) tumor cells

    International Nuclear Information System (INIS)

    Wallen, C.A.; Gutierrez, R.H.

    1987-01-01

    Alteration of the physiological environment of Q 66 and 67 mouse mammary tumor cells by placing them in either fresh, complete medium or a balanced salt solution supplemented with 24 mM glucose resulted in a significant increase in the time at 45 0 C necessary to measure cytotoxicity. The degree of increased resistance was dependent on the solution used to change the environment and the length of time the cells were allowed to equilibrate in this new environment. The aim of the present study is to determine if alterations in the Q cell environment has significant effects on the expression of thermotolerance. Pure P and Q cell populations of both 66 and 67 cell lines are exposed continuously to either 42 or 43 0 C and assayed for colony formation at various times for the development of thermotolerance. The comparison of thermotolerance development both in terms of time course and extent are measured in Q cells under 5 conditions: 1) normal, depleted medium (pH 6.8), 2) fresh, complete medium (pH 7.2), 3) balanced salt solution with 24 mM glucose (pH 7.2), 4) balanced salt solution with no glucose (pH 7.2), and 5) depleted medium supplemented with fresh serum (pH 6.8). These data have implications for the importance of Q cells in determining the outcome of clinical hyperthermia and the role of other stressors on the expression of thermotolerance

  16. 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

  17. Prevalence, antimicrobial resistance and risk factors for Campylobacter colonising dogs and cats in Greece

    Directory of Open Access Journals (Sweden)

    T. Lazou

    2017-09-01

    Full Text Available The study was conducted to determine the prevalence, antimicrobial resistance and risk factors for Campylobacter colonising dogs and cats in Greece. Faecal specimens were collected from 181 dogs and 132 cats. Culture methods were applied to detect Campylobacter spp. and a multiplex PCR assay to identify the isolates. The prevalence of Campylobacter spp. was 3.8% in dogs and 12.1% in cats. The most frequently identified Campylobacter species in dogs was C. jejuni (57.1% followed by C. coli (42.9%. All feline isolates were identified as C. jejuni apart from one isolate that was characterised as Campylobacter-like organism. Gender, age, breed, life style, diarrhoea and type of diet of dogs and cats did not significantly correlate (P>0.05 with Campylobacter isolation. Possible predictors regarding Campylobacter presence in dogs and cats were assessed by binary logistic regression. A tendency towards higher risk for Campylobacter contamination was observed in dogs consuming a homemade diet and in outdoor cats. Disk diffusion method revealed that all Campylobacter isolates exhibited susceptibility to erythromycin, gentamicin and streptomycin. Contrariwise, 66.7% of canine isolates were resistant concurrently to tetracycline and quinolones and 59.0%, 13.6% and 4.5% of feline isolates were resistant to quinolones, quinolones along with tetracycline and tetracycline alone, respectively

  18. Comparison of Antimicrobial Susceptibility of Campylobacter Strains Isolated from Food Samples and Patients with Diarrhea.

    Science.gov (United States)

    Bakhshi, Bita; Naseri, Amin; Alebouyeh, Masoud

    2016-01-01

    Campylobacter infections may lead to serious conditions, including septicemia or other invasive forms of the disease, which require rapid and accurate laboratory diagnosis and subsequently appropriate antimicrobial therapy. The aim of this study was to compare the species distribution and antimicrobial susceptibility pattern of Campylobacter spp. strains isolated from patients and food samples. Biochemical identification was performed on 15 clinical and 30 food isolates of Campylobacter recovered onto Brucella agar containing 5% sheep blood. PCR was carried out to confirm the identity of Campylobacter spp. using primers for cadF, hipO, and asp genes of Campylobacter. To determine antibiotic sensitivity of isolates, Kirby-Bauer assay was carried out using 16 different antibiotic discs. PCR assay and biochemical tests confirmed all 45 isolates as Campylobacter: 20 (44.44%) as C. jujeni, 10 (22.22%) as C. coli, and 15 (33.34%) as other Campylobacter strains. The maximum resistance was observed to cefotaxime and imipenem (each 86.49%) and the maximum sensitivity to erythromycin (48.65%). C. jujeni is dominant among isolates from clinical and food samples. In addition, tetracycline remains the first-line therapeutic agent against Campylobacter infections in Iran.

  19. Active Hexose Correlated Compound Extends the Lifespan and Increases the Thermotolerance of Nematodes

    Directory of Open Access Journals (Sweden)

    Tetsuya Okuyama

    2013-06-01

    Full Text Available ABSTRACTBackground: Active hexose correlated compound (AHCC is the extract from cultured mycelia of Lentinula edodes, a species of Basidiomycetes mushroom. AHCC contains various polysaccharides, including partially acylated -1,4-glucan, which is one of its major constituents. The application of AHCC has been markedly increased in complementary and alternative medicine as a functional food because AHCC improved the prognosis of postoperative hepatocellular carcinoma patients. AHCC has anti-inflammatory and antioxidant effects, such as the suppression of nitric oxide production in hepatocytes. AHCC might affect resistance to environmental stress, which is assumed to play a pivotal role in the longevity of many organisms.Objective: To investigate the effect of AHCC on longevity, we measured the lifespan of the nematode Caenorhabditis elegans, a model animal that is widely used to assess longevity. We also examined the effect of AHCC on resistance to heat stress, i.e., thermotolerance.Methods: The lifespan of C. elegans animals grown on media in the absence or presence of AHCC at 20°C was evaluated. Thermotolerance assays were performed at 35°C, the restrictive temperature of the animals. The effects of AHCC on lifespan and thermotolerance were analyzed with longevity mutants. Expression levels of stress-related genes, including heat shock genes, were measured by strand-specific reverse transcription-polymerase chain reaction after heat shock.Results: Wild-type C. elegans animals exhibited a longer mean lifespan by up to 10% in the presence of AHCC in the growth media than animals in the absence of AHCC. Furthermore, AHCC markedly increased thermotolerance at 35°C. Epistasis analyses showed that lifespan extension by AHCC at least partly required two longevity-promoting transcription factors: DAF-16 (C. elegans homolog of FOXO and HSF-1 (C. elegans homolog of heat shock transcription factor 1. After heat shock, AHCC activated the transcription

  20. Flying insects and Campylobacter

    DEFF Research Database (Denmark)

    Hald, Birthe; Sommer, Helle Mølgaard; Skovgård, Henrik

    organisms, which may collect on their bodies or survive passage through the fly gut. Campylobacter and other pathogens are then easily transferred to other surfaces, for instance peoples food – or to broiler houses where they may be swallowed by chickens or contaminate the environment. On a large material...... period was rather short, as even high doses of Campylobacter remained viable for less than 24 hours in flies, when they were incubated at temperatures from 20 ºC and higher. Lower temperatures are less- or irrelevant, as flies become slow or immobile below 15-20 ºC....

  1. Biofuels. Altered sterol composition renders yeast thermotolerant

    DEFF Research Database (Denmark)

    Caspeta, Luis; Chen, Yun; Ghiaci, Payam

    2014-01-01

    Ethanol production for use as a biofuel is mainly achieved through simultaneous saccharification and fermentation by yeast. Operating at ≥40°C would be beneficial in terms of increasing efficiency of the process and reducing costs, but yeast does not grow efficiently at those temperatures. We used...... adaptive laboratory evolution to select yeast strains with improved growth and ethanol production at ≥40°C. Sequencing of the whole genome, genome-wide gene expression, and metabolic-flux analyses revealed a change in sterol composition, from ergosterol to fecosterol, caused by mutations in the C-5 sterol...... desaturase gene, and increased expression of genes involved in sterol biosynthesis. Additionally, large chromosome III rearrangements and mutations in genes associated with DNA damage and respiration were found, but contributed less to the thermotolerant phenotype....

  2. 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.

  3. Campylobacter jejuni γ-glutamyltranspeptidase Activity Assay

    NARCIS (Netherlands)

    van der Stel, A.|info:eu-repo/dai/nl/12050200; Wösten, M.M.S.M.|info:eu-repo/dai/nl/164307249

    2016-01-01

    The enzyme γ-glutamyltranspeptidase (GGT, EC 2.3.2.2) is highly conserved among eukaryotic and prokaryotic organisms (Heisterkamp et al., 2008) and has a key function in glutathione metabolism. Although the enzyme is highly conserved and found throughout organisms ranging from bacteria to plants and

  4. Campylobacter hominis sp nov., from the human gastrointestinal tract

    DEFF Research Database (Denmark)

    Lawson, A.J.; On, Stephen L.W.; Logan, J.M.J.

    2001-01-01

    identified by a genus and taxon-specific PCR assay, and 16S rDNA nucleotide sequence analysis was carried out. All isolates exhibited the typical Campylobacter characteristics of being non-fermentative, oxidase-positive, catalase-negative and Gram-negative. Unusually, however, they were straight rods lacking...

  5. 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.

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

    Directory of Open Access Journals (Sweden)

    Jing eHan

    2012-03-01

    Full Text Available The prevalence of fluoroquinolone (FQ-resistant Campylobacter has become a concern for public health. To facilitate the control of FQ-resistant Campylobacter, it is necessary to understand the impact of FQ resistance on the fitness of Campylobacter in its natural hosts as understanding fitness will help to determine and predict the persistence of FQ-resistant Campylobacter. Previously it was shown that acquisition of resistance to FQ antimicrobials enhanced the in vivo fitness of FQ-resistant Campylobacter. In this study, we confirmed the role of the Thr-86-Ile mutation in GyrA in modulating Campylobacter fitness by reverting the mutation to the wild-type allele, which resulted in the loss of the fitness advantage. Additionally, we determined if the resistance-conferring GyrA mutations alter the enzymatic function of the DNA gyrase. Recombinant wild-type gyrase and mutant gyrases with three different types of mutations (Thr-86-Ile, Thr-86-Lys, and Asp-90-Asn, which are associated with FQ resistance in Campylobacter, were generated in E. coli and compared for their supercoiling activities using an in vitro assay. The mutant gyrase with the Thr-86-Ile change showed a greatly reduced supercoiling activity compared with the wild-type gyrase, while other mutant gyrases did not show an altered supercoiling. Furthermore, we measured DNA supercoiling within Campylobacter cells using a reporter plasmid. Consistent with the results from the in vitro supercoiling assay, the FQ-resistant mutant carrying the Thr-86-Ile change in GyrA showed much less DNA supercoiling than the wild-type strain and the mutant strains carrying other mutations. Together, these results indicate that the Thr-86-Ile mutation, which is predominant in clinical FQ-resistant Campylobacter, modulates DNA supercoiling homeostasis in FQ-resistant Campylobacter.

  7. Elucidation of thermotolerance diversity in cotton (Gossypium hirsutum L.) using physio-molecular approaches.

    Science.gov (United States)

    Rana, R M; Khan, S H; Ali, Z; Khan, A I; Khan, I A

    2011-06-14

    Cotton (Gossypium hirsutum) is an important cash crop, but high temperature during its growing season is one of the major factors that limit its productivity. This problem compels plant breeders to breed for heat tolerance, which can help to overcome this challenge. It is very important to make a comprehensive screening of heat-tolerant genotypes so that only the best are chosen. Here we report the combined use of several techniques that can help breeders to screen their germplasm. Twelve cultivated cotton genotypes were evaluated for thermotolerance, using assays that included electrolyte leakage, chlorophyll accumulation and protein profiling, as well as RAPDs to assess genetic diversity. Two genotypes (B-557 and NIAB-78) showed tolerant behavior in three thermotolerance assays. RAPD analysis results showed maximum similarity in a range of 86.7-66.7% between the genotypes MNH-554 and CIM-443. We conclude that combined use should be made of relative electrolyte leakage, chlorophyll stability and differential display with SDS-PAGE to aid in screening for stress tolerance. RAPD-based diversity analysis will further help to improve the efficiency of breeding programs.

  8. Antibiotic susceptibility pattern and genotyping of campylobacter species isolated from children suffering from gastroenteritis.

    Science.gov (United States)

    Abd El-Baky, R M; Sakhy, M; Gad, G F M

    2014-01-01

    To study the prevalence and the antimicrobial resistance of campylobacter species isolated from children suffering from gastroenteritis . A total of 125 stool samples were collected from children with gastroenteritis. The identification of isolates was performed with conventional methods as well as with molecular methods based on 16SrRNA species-specific gene amplification by PCR and product analysis. Resistance pattern of the isolated strains was determined using agar dilution method. Conventional methods including sodium hippurate hydrolysis revealed that 12 (9.6%) samples were positive for Campylobacter species. Ten out of 12 Campylobacter spp. were identified as Campylobacter jejuni and 2 as Campylobacter coli but PCR assay revealed that five samples only were positive for Campylobacter and all were C. jejuni. Antimicrobial susceptibility to 10 antimicrobials was performed and all isolates (five isolates of C. jejuni) were susceptible to chloramphenicol, gentamicin and amikacin but all were resistant to ceftriaxone. PCR assay method allows reliable detection of C. jejuni. C. jejuni was the most prevalent Campylobacter species. Gentamicin, amikacin and chloramphenicol were the most effective antibiotic.

  9. Antibiotic susceptibility pattern and genotyping of campylobacter species isolated from children suffering from gastroenteritis

    Directory of Open Access Journals (Sweden)

    R M Abd El-Baky

    2014-01-01

    Full Text Available Purpose: To study the prevalence and the antimicrobial resistance of campylobacter species isolated from children suffering from gastroenteritis . Materials and Methods: A total of 125 stool samples were collected from children with gastroenteritis. The identification of isolates was performed with conventional methods as well as with molecular methods based on 16SrRNA species-specific gene amplification by PCR and product analysis. Resistance pattern of the isolated strains was determined using agar dilution method. Results: Conventional methods including sodium hippurate hydrolysis revealed that 12 (9.6% samples were positive for Campylobacter species. Ten out of 12 Campylobacter spp. were identified as Campylobacter jejuni and 2 as Campylobacter coli but PCR assay revealed that five samples only were positive for Campylobacter and all were C. jejuni. Antimicrobial susceptibility to 10 antimicrobials was performed and all isolates (five isolates of C. jejuni were susceptible to chloramphenicol, gentamicin and amikacin but all were resistant to ceftriaxone. Conclusion: PCR assay method allows reliable detection of C. jejuni. C. jejuni was the most prevalent Campylobacter species. Gentamicin, amikacin and chloramphenicol were the most effective antibiotic.

  10. Thermotolerant cyclamen with reduced acrolein and methyl vinyl ketone.

    Science.gov (United States)

    Kai, Hiroomi; Hirashima, Keita; Matsuda, Osamu; Ikegami, Hidetoshi; Winkelmann, Traud; Nakahara, Takao; Iba, Koh

    2012-06-01

    Reduced levels of trienoic fatty acids (TAs) in chloroplast membranes induce thermotolerance in several plant species, but the underlying mechanisms remain unclear. TA peroxidation in plant cell membranes generates cytotoxic, TA-derived compounds containing α,β-unsaturated carbonyl groups. The relationship between low TA levels and the amounts of cytotoxic TA-derived compounds was examined using thermotolerant transgenic cyclamen (Cyclamen persicum Mill.) with low TA contents. Changes in the levels of the cytotoxic TA-derived acrolein (ACR), methyl vinyl ketone (MVK), (E)-2-hexenal, 4-hydroxy-2-nonenal, and malondialdehyde were analysed in the leaf tissues of wild-type (WT) and thermotolerant transgenic cyclamen under heat stress. Levels of ACR and MVK in the WT increased in parallel with the occurrence of heat-induced tissue damage, whereas no such changes were observed in the thermotolerant transgenic lines. Furthermore, exogenous ACR and MVK infiltrated into leaves to concentrations similar to those observed in heat-stressed WT leaves caused similar disease symptoms. These results suggest that thermotolerance in transgenic cyclamen depends on reduced production rates of ACR and MVK under heat stress, due to the low level of TAs in these plants.

  11. Thermotolerant yeasts and application for ethanol production

    Directory of Open Access Journals (Sweden)

    To-on, N.

    2007-07-01

    Full Text Available A total of 70 thermotolerant yeast strains were isolated at 40oC from 145 samples including fruit, leaves, flowers, soils and oil-palm fruits. Six isolates showed maximum growth at 40oC within 18 h. Three isolates (MIY1, MIY48 and MIY57 were selected based on their ability to ferment glucose and sucrose rapidly (24 h and showed the maximum temperature for growth at 42oC but it was good at 40oC. MIY57 produced 4.6% (v/v ethanol at 40oC from a medium containing 15% glucose. The optimum cultivation conditions for growth and ethanol production of MIY57 was 5% inoculum into the fermentation medium containing 15% glucose and 1% yeast extract with initial pH of 4.5 on a shaking incubator at 150 rpm at 40oC. MIY57, under these conditions, produced maximum ethanol of 5.0% (v/v after 48 h incubation while S. cerevisiae TISTR 5048 produced only 3.7% (v/v. Maximum cell dry weight was 7.2 g/L (at 18 h, again much higher than that of S. cerevisiae TISTR 5048 (4.1 g/L. Based on morphological, physiological and molecular studies, this strain (MIY57 was identified as Saccharomyces cerevisiae.

  12. 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.

  13. Campylobacter Risk Analysis

    DEFF Research Database (Denmark)

    Nauta, Maarten

    In several countries quantitative microbiological risk assessments (QMRAs) have been performed for Campylobacter in chicken meat. The models constructed for this purpose provide a good example of the development of QMRA in general and illustrate the diversity of available methods. Despite...... the differences between the models, the most prominent conclusions of the QMRAs are similar. These conclusions for example relate to the large risk of highly contaminated meat products and the insignificance of contamination from Campylobacter positive flocks to negative flocks during slaughter and processing....... Nonetheless, there seems to be a discrepancy between model predictions and the accumulating microbiological data. For example, a recent study in the Netherlands showed that model predictions on the efficacy of “testing and scheduling” of broiler flocks as a control strategy, could not be confirmed...

  14. Campylobacter hyointestinalis subsp hyointestinalis, a common Campylobacter species in reindeer

    DEFF Research Database (Denmark)

    Hanninen, M.L.; Sarelli, L.; Sukura, A.

    2002-01-01

    Aims: To study the prevalence of Campylobacter spp. in the faecal material of reindeer, and to identify the isolates by means of a polyphasic approach. In addition, to study the genetic diversity of Camp. hyointestinalis subsp. hyointestinalis reindeer isolates by pulsed-field gel electrophoresis...... slaughterhouses. Samples were cultured by methods suitable for isolation of fastidious Campylobacter species. Of all samples, 6% (24/399) were Campylobacter-positive. Phenotypic characteristics, SDS-PAGE protein patterns, dot blot DNA-DNA hybridization, 23S rDNA restriction fragment polymorphism analysis and PFGE...... identified the isolates as Camp. hyointestinalis subsp. kyointestinalis. Conclusions: Campylobacter hyointestinalis subsp. hyointestinalis was the only Campylobacter species isolated from reindeer in this study. The isolates showed high genomic diversity in PFGE with the restriction enzymes SmaI and Kpn...

  15. Gastroenteritis caused by Campylobacter concisus.

    Science.gov (United States)

    Hess, D L J; Pettersson, A M; Rijnsburger, M C; Herbrink, P; van den Berg, H P; Ang, C W

    2012-05-01

    We describe a case of gastroenteritis caused by Campylobacter concisus. The pathogenic potential of C. concisus has yet to be elucidated. Recent studies indicate an association with enteric disease in immunocompromised patients and inflammatory bowel disease in children. Molecular identification methods may be necessary for identifying certain Campylobacter species because of phenotypic similarity. © 2012 SGM

  16. High salinity conveys thermotolerance in the coral model Aiptasia

    KAUST Repository

    Gegner, Hagen M.

    2017-12-15

    The endosymbiosis between dinoflagellate algae of the genus Symbiodinium and stony corals provides the foundation of coral reef ecosystems. Coral bleaching, the expulsion of endosymbionts from the coral host tissue as a consequence of heat or light stress, poses a threat to reef ecosystem functioning on a global scale. Hence, a better understanding of the factors contributing to heat stress susceptibility and tolerance is needed. In this regard, some of the most thermotolerant corals also live in particularly saline habitats, but possible effects of high salinity on thermotolerance in corals are anecdotal. Here we test the hypothesis that high salinity may lead to increased thermotolerance. We conducted a heat stress experiment at low, intermediate, and high salinities using a set of host-endosymbiont combinations of the coral model Aiptasia. As expected, all host-endosymbiont combinations showed reduced photosynthetic efficiency and endosymbiont loss during heat stress, but the severity of bleaching was significantly reduced with increasing salinities for one of the host-endosymbiont combinations. Our results show that higher salinities can convey increased thermotolerance in Aiptasia, although this effect seems to be dependent on the particular host strain and/or associated symbiont type. This finding may help explain the extraordinarily high thermotolerance of corals in high salinity environments such as the Red Sea and the Persian/Arabian Gulf and provides novel insight regarding factors that contribute to thermotolerance. Since our results are based on a salinity effect in symbiotic sea anemones, it remains to be determined whether this salinity effect can also be observed in stony corals.

  17. Real-time PCR detection of Campylobacter spp. In free-ranging mountain gorillas (Gorilla beringei beringei).

    Science.gov (United States)

    Whittier, Christopher A; Cranfield, Michael R; Stoskopf, Michael K

    2010-07-01

    Health monitoring of wildlife populations can greatly benefit from rapid, local, noninvasive molecular assays for pathogen detection. Fecal samples collected from free-living Virunga mountain gorillas (Gorilla beringei beringei) between August 2002 and February 2003 were tested for Campylobacter spp. DNA using a portable, real-time polymerase chain reaction (PCR) instrument. A high prevalence of Campylobacter spp. was detected in both individually identified (22/26=85%) and nest-collected samples (68/114=59.6%), with no statistically significant differences among different gorilla sexes or age classes or between tourist-visited versus research gorilla groups. The PCR instrument was able to discriminate two distinct groups of Campylobacter spp. in positive gorilla samples based on the PCR product fluorescent-probe melting profiles. The rare type (6/90 positives, 7%, including three mixed cases) matched DNA sequences of Campylobacter jejuni and was significantly associated with abnormally soft stools. The more common type of positive gorilla samples (87/90 positives, 97%) were normally formed and contained a Campylobacter sp. with DNA matching no published sequences. We speculate that the high prevalence of Campylobacter spp. detected in gorilla fecal samples in this survey mostly reflects previously uncharacterized and nonpathogenic intestinal flora. The real-time PCR assay was more sensitive than bacterial culture with Campylobacter-specific media and commercially available, enzyme immunoassay tests for detecting Campylobacter spp. in human samples.

  18. 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.

  19. 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.

  20. [Evaluation of the role of Campylobacter spp. in the occurrence of foodborne diseases and modern methods to detect the pathogen].

    Science.gov (United States)

    Efimochkina, N R

    2015-01-01

    to quantify present in foods of thermotolerant Campylobacter, including C. jejuni.

  1. Effect of thermotolerance on thermal radiosensitization in hepatoma cells

    International Nuclear Information System (INIS)

    van Rijn, J.; van den Berg, J.; Schamhart, D.H.J.; van Wijk, R.

    1984-01-01

    The interaction between hyperthermia and X irradiation was determined in cultured Reuber H35 hepatoma cells with different states of thermosensitivity. Incubation at 41 0 C followed by 4-Gy X rays resulted after 2 hr in a stabilization of cell survival for heat or heat plus X rays, with a maximum synergism factor of 1.6. Thermotolerance did not develop during incubation at 41.7 or 42.5 0 C. When heat treatment of cells was followed by irradiation, the synergism factor for thermal radiosensitization increased with both the amount of thermal cell killing and the amount of X-ray cell killing; the influence of thermal exposure on the synergism factor was greater than that of the X-ray dose. Cells were made thermotolerant either by incubation at 42.5 0 C for 30 or 60 min followed by an interval at 37 0 C, or by continuous incubation at 41 0 C. In both cases thermotolerance was measured by incubation at 42.5 0 C. No difference was observed between the maximum thermotolerance achieved with both methods. When cells were irradiated in addition to the second heat treatment, thermal radiosensitization was strongly reduced concomitant with the decreased sensitivity to killing by heat

  2. Polyphasic identification of a new thermotolerant species of lactic ...

    African Journals Online (AJOL)

    Two thermotolerant and desiccation tolerant lactic acid bacteria (TDLAB) were pointed out from twenty isolated strains from soils and dried chicken faeces. Samples were collected in poultry farms in the vicinity of Dakar, Senegal (West Africa). The two new isolates were called Sp.4 (Sp.4=CWBI-B534=LMG7278) and Sp.20 ...

  3. Induction of thermotolerance through heat acclimation and salicylic ...

    African Journals Online (AJOL)

    High temperature stress is the second most important stress, which can strike crop plants at any time and impose severe limitations on crop growth and development. Developing crop plants with improved thermotolerance can mitigate the adverse effects of heat stress. However, a thorough understanding of physiological ...

  4. Isolation and characterization of thermotolerant ethanol-fermenting ...

    African Journals Online (AJOL)

    Thermotolerant yeasts, which are expected to be applicable for high-temperature fermentation as an economical process, were isolated from four provinces in Laos. Of these yeasts, five isolates exhibited stronger fermentation abilities in a 16% sugars-containing medium of glucose, sucrose, sugarcane or molasses at 40°C ...

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

    Directory of Open Access Journals (Sweden)

    Vardund T

    2006-06-01

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

  6. 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

  7. 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.

  8. Identification of Heat Shock Transcription Factor Genes Involved in Thermotolerance of Octoploid Cultivated Strawberry.

    Science.gov (United States)

    Liao, Wan-Yu; Lin, Lee-Fong; Jheng, Jing-Lian; Wang, Chun-Chung; Yang, Jui-Hung; Chou, Ming-Lun

    2016-12-17

    Heat shock transcription factors (HSFs) are mainly involved in the activation of genes in response to heat stress as well as other abiotic and biotic stresses. The growth, development, reproduction, and yield of strawberry are strongly limited by extreme temperatures and droughts. In this study, we used Illumina sequencing and obtained transcriptome data set from Fragaria × ananassa Duchessne cv. Toyonoka. Six contigs and three unigenes were confirmed to encode HSF proteins (FaTHSFs). Subsequently, we characterized the biological functions of two particularly selected unigenes, FaTHSFA2a and FaTHSFB1a , which were classified into class A2 and B HSFs, respectively. Expression assays revealed that FaTHSFA2a and FaTHSFB1a expression was induced by heat shock and correlated well with elevated ambient temperatures. Overexpression of FaTHSFA2a and FaTHSFB1a resulted in the activation of their downstream stress-associated genes, and notably enhanced the thermotolerance of transgenic Arabidopsis plants. Besides, both FaTHSFA2a and FaTHSFB1a fusion proteins localized in the nucleus, indicating their similar subcellular distributions as transcription factors. Our yeast one-hybrid assay suggested that FaTHSFA2a has trans-activation activity, whereas FaTHSFB1a expresses trans-repression function. Altogether, our annotated transcriptome sequences provide a beneficial resource for identifying most genes expressed in octoploid strawberry. Furthermore, HSF studies revealed the possible insights into the molecular mechanisms of thermotolerance, thus rendering valuable molecular breeding to improve the tolerance of strawberry in response to high-temperature stress.

  9. Prevalence and Antimicrobial Resistance of Campylobacter Isolated from Dressed Beef Carcasses and Raw Milk in Tanzania.

    Science.gov (United States)

    Kashoma, Isaac P; Kassem, Issmat I; John, Julius; Kessy, Beda M; Gebreyes, Wondwossen; Kazwala, Rudovick R; Rajashekara, Gireesh

    2016-01-01

    Campylobacter species are commonly transmitted to humans through consumption of contaminated foods such as milk and meat. The aim of this study was to investigate the prevalence, antimicrobial resistance, and genetic determinants of resistance of Campylobacter isolated from raw milk and beef carcasses in Tanzania. The antimicrobial resistance genes tested included blaOXA-61 (ampicillin), aph-3-1 (aminoglycoside), tet(O) (tetracycline), and cmeB (multi-drug efflux pump). The prevalence of Campylobacter was 9.5% in beef carcasses and 13.4% in raw milk, respectively. Using multiplex-polymerase chain reaction (PCR), we identified 58.1% of the isolates as Campylobacter jejuni, 30.7% as Campylobacter coli, and 9.7% as other Campylobacter spp. One isolate (1.6%) was positive for both C. jejuni and C. coli specific PCR. Antimicrobial susceptibility testing using the disk diffusion assay and the broth microdilution method showed resistance to: ampicillin (63% and 94.1%), ciprofloxacin (9.3% and 11.8%), erythromycin (53.7% and 70.6%), gentamicin (0% and 15.7%), streptomycin (35.2% and 84.3%), and tetracycline (18.5% and 17.7%), respectively. Resistance to azithromycin (42.6%), nalidixic acid (64.8%), and chloramphenicol (13%) was determined using the disk diffusion assay only, while resistance to tylosin (90.2%) was quantified using the broth microdilution method. The blaOXA-61 (52.6% and 28.1%), cmeB (26.3% and 31.3%), tet(O) (26.3% and 31.3%), and aph-3-1 (5.3% and 3.0%) were detected in C. coli and C. jejuni. These findings highlight the extent of antimicrobial resistance in Campylobacter occurring in important foods in Tanzania. The potential risks to consumers emphasize the need for adequate control approaches, including the prudent use of antimicrobials to minimize the spread of antimicrobial-resistant Campylobacter.

  10. Antimicrobial Resistance and Genotypic Diversity of Campylobacter Isolated from Pig, Dairy and Beef Cattle in Tanzania

    Directory of Open Access Journals (Sweden)

    Isaac eKashoma

    2015-11-01

    Full Text Available Foodborne Campylobacter infections pose a serious threat to public health worldwide. However, the occurrence and characteristics of Campylobacter in food animals and products remain largely unknown in Tanzania. The objective of this study was to determine the prevalence, antibiotic resistance, and genetic profiles (sequence types, STs of Campylobacter isolated from feces of pigs and dairy and beef cattle in Tanzania. Overall, 259 (~ 30% of 864 samples were positive for Campylobacter spp, which were detected in 32.5%, 35.4%, and 19.6% of the pig, dairy, and beef cattle samples, respectively. Multiplex PCR analysis identified 64.5% and 29.3% of the Campylobacter isolates as C. coli and C. jejuni, respectively. The majority (91.9% of the isolates from pig samples were identified as C. coli, while C. jejuni accounted for 65.5% of the isolates from cattle. Antimicrobial susceptibility testing using the disk diffusion assay and the broth microdilution method revealed resistance to: ampicillin (70% and 76%, gentamicin (1.8% and 12.6%, respectively, streptomycin (65.8% and 74.8%, erythromycin (41.4% and 48.7%, tetracycline (18.9% and 23.4%, and ciprofloxacin (14.4% and 7.2%. Resistance to nalidixic acid (39.6%, azithromycin (13.5%, and chloramphenicol (4.5% was determined using the disk diffusion assay only, while resistance to tylosin (38.7% was quantified using the broth microdilution method. Multilocus sequence typing of 111 Campylobacter isolates resulted in the identification of 48 STs (26 C. jejuni and 22 C. coli of which 7 were novel (6 C. jejuni and 1 C. coli. Taken together, this study revealed the high prevalence, genetic diversity and antimicrobial resistance of Campylobacter in important food animals in Tanzania, which highlights the urgent need for the surveillance and control of Campylobacter in this country.

  11. A pre-enrichment step is essential for detection of Campylobacter sp. in turbid pond water.

    Science.gov (United States)

    Abulreesh, H H; Paget, T A; Goulder, R

    2014-06-01

    This work aimed to detect Campylobacter species from naturally contaminated turbid pond water by PCR. A total of 16 water samples were collected from a turbid village pond. Four methods of DNA extraction were applied to centrifuge pellets from eight 100 ml pond water samples prior to attempted detection of Campylobacter by PCR without an enrichment step. These methods were (1) Tris-HCl and sodium dodecyl sulfate followed by phenol:chloroform:isoamylalcohol extraction followed by treatment with DNA clean up kit, (2) proteinase K, (3) Chelex® 100, and (4) boiling. The other eight pond water samples (10 ml and 100 ml) were filtered and filters were incubated overnight in Preston enrichment broth. The centrifuge pellets obtained from enrichment cultures were treated by proteinase K for DNA extraction. Primers CF03 and CF04 for the flagellin genes (flaA and flaB) of Campylobacter jejuni and Campylobacter coli were used for amplifying the extracted DNA. The DNA extracted from eight-100 ml pond water samples that were not subject to selective enrichment was never amplified with primers CF03 and CF04, hence Campylobacter was not detected. In contrast, the DNA that was from samples that were subjected to a selective enrichment step in Preston broth prior to PCR assay always gave amplified bands of 340-380 bp, therefore the presence of Campylobacter was confirmed. Detection of campylobacters from naturally contaminated, turbid, environmental water may not be feasible by direct PCR assay because of low numbers and the presence of high concentration of humic matter and other PCR inhibitors. The enrichment of water samples in selective broth, however, facilitated PCR detection of Campylobacter probably by increasing cell number and by diluting PCR inhibitors.

  12. Multicenter Evaluation of Clinical Diagnostic Methods for Detection and Isolation of Campylobacter spp. from Stool.

    Science.gov (United States)

    Fitzgerald, Collette; Patrick, Mary; Gonzalez, Anthony; Akin, Joshua; Polage, Christopher R; Wymore, Kate; Gillim-Ross, Laura; Xavier, Karen; Sadlowski, Jennifer; Monahan, Jan; Hurd, Sharon; Dahlberg, Suzanne; Jerris, Robert; Watson, Renee; Santovenia, Monica; Mitchell, David; Harrison, Cassandra; Tobin-D'Angelo, Melissa; DeMartino, Mary; Pentella, Michael; Razeq, Jafar; Leonard, Celere; Jung, Carrianne; Achong-Bowe, Ria; Evans, Yaaqobah; Jain, Damini; Juni, Billie; Leano, Fe; Robinson, Trisha; Smith, Kirk; Gittelman, Rachel M; Garrigan, Charles; Nachamkin, Irving

    2016-05-01

    The use of culture-independent diagnostic tests (CIDTs), such as stool antigen tests, as standalone tests for the detection of Campylobacter in stool is increasing. We conducted a prospective, multicenter study to evaluate the performance of stool antigen CIDTs compared to culture and PCR for Campylobacter detection. Between July and October 2010, we tested 2,767 stool specimens from patients with gastrointestinal illness with the following methods: four types of Campylobacter selective media, four commercial stool antigen assays, and a commercial PCR assay. Illnesses from which specimens were positive by one or more culture media or at least one CIDT and PCR were designated "cases." A total of 95 specimens (3.4%) met the case definition. The stool antigen CIDTs ranged from 79.6% to 87.6% in sensitivity, 95.9 to 99.5% in specificity, and 41.3 to 84.3% in positive predictive value. Culture alone detected 80/89 (89.9% sensitivity) Campylobacter jejuni/Campylobacter coli-positive cases. Of the 209 noncases that were positive by at least one CIDT, only one (0.48%) was positive by all four stool antigen tests, and 73% were positive by just one stool antigen test. The questionable relevance of unconfirmed positive stool antigen CIDT results was supported by the finding that noncases were less likely than cases to have gastrointestinal symptoms. Thus, while the tests were convenient to use, the sensitivity, specificity, and positive predictive value of Campylobacter stool antigen tests were highly variable. Given the relatively low incidence of Campylobacter disease and the generally poor diagnostic test characteristics, this study calls into question the use of commercially available stool antigen CIDTs as standalone tests for direct detection of Campylobacter in stool. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

  13. Antimicrobial Resistance and Genotypic Diversity of Campylobacter Isolated from Pigs, Dairy, and Beef Cattle in Tanzania

    Science.gov (United States)

    Kashoma, Isaac P.; Kassem, Issmat I.; Kumar, Anand; Kessy, Beda M.; Gebreyes, Wondwossen; Kazwala, Rudovick R.; Rajashekara, Gireesh

    2015-01-01

    Foodborne Campylobacter infections pose a serious threat to public health worldwide. However, the occurrence and characteristics of Campylobacter in food animals and products remain largely unknown in Tanzania. The objective of this study was to determine the prevalence, antibiotic resistance, and genetic profiles (sequence types, STs) of Campylobacter isolated from feces of pigs and dairy and beef cattle in Tanzania. Overall, 259 (~30%) of 864 samples were positive for Campylobacter spp, which were detected in 32.5, 35.4, and 19.6% of the pig, dairy, and beef cattle samples, respectively. Multiplex PCR analysis identified 64.5 and 29.3% of the Campylobacter isolates as C. coli and C. jejuni, respectively. The majority (91.9%) of the isolates from pig samples were identified as C. coli, while C. jejuni accounted for 65.5% of the isolates from cattle. Antimicrobial susceptibility testing using the disk diffusion assay and the broth microdilution method revealed resistance to: ampicillin (Amp) (70.3% and 75.7%, respectively), gentamicin (Gen) (1.8% and 12.6%), streptomycin (Str) (65.8 and 74.8%), erythromycin (Ery) (41.4 and 48.7%), tetracycline (Tet) (18.9 and 23.4%), and ciprofloxacin (Cip) (14.4 and 7.2%). Resistance to nalidixic acid (Nal) (39.6%), azithromycin (Azm) (13.5%), and chloramphenicol (Chl) (4.5%) was determined using the disk diffusion assay only, while resistance to tylosin (Tyl) (38.7%) was quantified using the broth microdilution method. Multilocus sequence typing of 111 Campylobacter isolates resulted in the identification of 48 STs (26 C. jejuni and 22 C. coli) of which seven were novel (six C. jejuni and one C. coli). Taken together, this study revealed the high prevalence, genetic diversity and antimicrobial resistance of Campylobacter in important food animals in Tanzania, which highlights the urgent need for the surveillance and control of Campylobacter in this country. PMID:26617582

  14. Campylobacter infection in children in Malawi is common and is frequently associated with enteric virus co-infections.

    Directory of Open Access Journals (Sweden)

    Jenifer Mason

    Full Text Available Campylobacter species are the most common cause of bacterial gastroenteritis in the developed world. However, comparatively few studies have determined the epidemiological features of campylobacteriosis in resource-poor settings.A total of 1,941 faecal specimens collected from symptomatic (diarrhoeic children and 507 specimens from asymptomatic (non-diarrhoeic children hospitalised in Blantyre, Malawi, between 1997 and 2007, and previously tested for the presence of rotavirus and norovirus, was analysed for C. jejuni and C. coli using a real time PCR assay.Campylobacter species were detected in 415/1,941 (21% of diarrhoeic children, with C. jejuni accounting for 85% of all cases. The median age of children with Campylobacter infection was 11 months (range 0.1-55 months, and was significantly higher than that for children with rotavirus and norovirus (6 months and 7 months respectively; P<0.001. Co-infection with either rotavirus or norovirus was noted in 41% of all cases in the diarrhoeic group. In contrast, the detection rate of Campylobacter in the non-diarrhoeic group was 14%, with viral co-infection identified in 16% of children with Campylobacter. There was no association between Campylobacter detection rate and season over the 10 year period.Using molecular detection methodology in hospitalised Malawian children, we have demonstrated a high prevalence of Campylobacter infection, with frequent viral co-infection. The burden of Campylobacter infection in young African children may be greater than previously recognised.

  15. Campylobacter bacteriophages and bacteriophage therapy.

    Science.gov (United States)

    Connerton, P L; Timms, A R; Connerton, I F

    2011-08-01

    Members of the genus Campylobacter are frequently responsible for human enteric disease with occasionally very serious outcomes. Much of this disease burden is thought to arise from consumption of contaminated poultry products. More than 80% of poultry in the UK harbour Campylobacter as a part of their intestinal flora. To address this unacceptably high prevalence, various interventions have been suggested and evaluated. Among these is the novel approach of using Campylobacter-specific bacteriophages, which are natural predators of the pathogen. To optimize their use as therapeutic agents, it is important to have a comprehensive understanding of the bacteriophages that infect Campylobacter, and how they can affect their host bacteria. This review will focus on many aspects of Campylobacter-specific bacteriophages including: their first isolation in the 1960s, their use in bacteriophage typing schemes, their isolation from the different biological sources and genomic characterization. As well as their use as therapeutic agents to reduce Campylobacter in poultry their future potential, including their use in bio-sanitization of food, will be explored. The evolutionary consequences of naturally occurring bacteriophage infection that have come to light through investigations of bacteriophages in the poultry ecosystem will also be discussed. © 2011 The Authors. Journal of Applied Microbiology © 2011 The Society for Applied Microbiology.

  16. Antimicrobial resistance mechanisms among Campylobacter.

    Science.gov (United States)

    Wieczorek, Kinga; Osek, Jacek

    2013-01-01

    Campylobacter jejuni and Campylobacter coli are recognized as the most common causative agents of bacterial gastroenteritis in the world. Humans most often become infected by ingesting contaminated food, especially undercooked chicken, but also other sources of bacteria have been described. Campylobacteriosis is normally a self-limiting disease. Antimicrobial treatment is needed only in patients with more severe disease and in those who are immunologically compromised. The most common antimicrobial agents used in the treatment of Campylobacter infections are macrolides, such as erythromycin, and fluoroquinolones, such as ciprofloxacin. Tetracyclines have been suggested as an alternative choice in the treatment of clinical campylobacteriosis but in practice are not often used. However, during the past few decades an increasing number of resistant Campylobacter isolates have developed resistance to fluoroquinolones and other antimicrobials such as macrolides, aminoglycosides, and beta-lactams. Trends in antimicrobial resistance have shown a clear correlation between use of antibiotics in the veterinary medicine and animal production and resistant isolates of Campylobacter in humans. In this review, the patterns of emerging resistance to the antimicrobial agents useful in treatment of the disease are presented and the mechanisms of resistance to these drugs in Campylobacter are discussed.

  17. Antimicrobial Resistance Mechanisms among Campylobacter

    Science.gov (United States)

    2013-01-01

    Campylobacter jejuni and Campylobacter coli are recognized as the most common causative agents of bacterial gastroenteritis in the world. Humans most often become infected by ingesting contaminated food, especially undercooked chicken, but also other sources of bacteria have been described. Campylobacteriosis is normally a self-limiting disease. Antimicrobial treatment is needed only in patients with more severe disease and in those who are immunologically compromised. The most common antimicrobial agents used in the treatment of Campylobacter infections are macrolides, such as erythromycin, and fluoroquinolones, such as ciprofloxacin. Tetracyclines have been suggested as an alternative choice in the treatment of clinical campylobacteriosis but in practice are not often used. However, during the past few decades an increasing number of resistant Campylobacter isolates have developed resistance to fluoroquinolones and other antimicrobials such as macrolides, aminoglycosides, and beta-lactams. Trends in antimicrobial resistance have shown a clear correlation between use of antibiotics in the veterinary medicine and animal production and resistant isolates of Campylobacter in humans. In this review, the patterns of emerging resistance to the antimicrobial agents useful in treatment of the disease are presented and the mechanisms of resistance to these drugs in Campylobacter are discussed. PMID:23865047

  18. Aspects of epidemiology of Campylobacter in poultry

    NARCIS (Netherlands)

    Jacobs-Reitsma, W.F.

    1997-01-01

    Campylobacter bacteria, which in humans cause infections with severe symptoms of diarrhoea, are mainly transmitted by food, especially poultry meat products. Several studies on Campylobacter colonization in breeders, laying hens, and broilers were carried out. Isolates were serotyped, using a

  19. Anti-Campylobacter Activities and Resistance Mechanisms of Natural Phenolic Compounds in Campylobacter

    OpenAIRE

    Klan?nik, Anja; Mo?ina, Sonja Smole; Zhang, Qijing

    2012-01-01

    BACKGROUND: Campylobacter is a major foodborne pathogen and alternative antimicrobials are needed to prevent or decrease Campylobacter contamination in foods or food producing animals. The objectives of this study are to define the anti-Campylobacter activities of natural phenolic compounds of plant origin and to determine the roles of bacterial drug efflux systems in the resistance to these natural phenolics in Campylobacter jejuni. METHODOLOGY/PRINCIPAL FINDINGS: Anti-Campylobacter activiti...

  20. 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.

  1. 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.

  2. Prevalence and antimicrobial resistance in Campylobacter from ...

    African Journals Online (AJOL)

    Poultry are recognized as a main reservoir of Campylobacter spp. However, longitudinal studies investigating the persistence of Campylobacter on broilers and retail chciekn meat in Tanzania are rare. The aim of the current work was to evaluate the prevalence and antimicrobial susceptibility of Campylobacter spp. isolated ...

  3. Kip, consument en Campylobacter: infectieziektebestrijding met consumentenvoorlichting

    NARCIS (Netherlands)

    Jonge, de R.; Fischer, A.R.H.; Nauta, M.J.

    2007-01-01

    In the Netherlands, Campylobacter is an important bacterial causative agent of gastroenteritis, with poultry as an important source. Measures aiming to reduce the number of Campylobacter on poultry meat however cannot guarantee the production of fresh meat free of Campylobacter. To minimize the

  4. Leaf thermotolerance in dry tropical forest tree species: relationships with leaf traits and effects of drought.

    Science.gov (United States)

    Sastry, Aniruddh; Guha, Anirban; Barua, Deepak

    2018-02-01

    Understanding how tropical trees will respond to extreme temperatures and drought is essential to predict how future increases in the severity, frequency and duration of extreme climatic events will affect tropical systems. In this study, we investigated leaf thermotolerance by quantifying the temperatures that resulted in a 50 % decrease in photosystem II function (T 50 ) in experimentally grown saplings of 12 tree species from a seasonally dry tropical forest. We examined the relationship of thermotolerance with leaf functional traits and photosynthetic rates. Additionally, we tested how water limitation altered thermotolerance within species, and examined the relationship between thermotolerance and drought tolerance among species. Thermotolerance ranged from 44.5 to 48.1 °C in the least and most thermotolerant species, respectively. The observed variation in thermotolerance indicates that the upper limits of leaf function are critically close to maximum temperatures in this region, and that these species will be vulnerable to, and differentially affected by, future warming. Drought increased temperature tolerance, and species that were more drought tolerant were also more thermotolerant. Importantly, thermotolerance was positively related to the key leaf functional trait-leaf mass per area (LMA), and congruent with this, negatively related to photosynthetic rates. These results indicate that more productive species with lower LMA and higher photosynthetic rates may be more vulnerable to heat and drought stress, and more likely to be negatively affected by future increases in extreme climatic events.

  5. Expression and thermotolerance of calreticulin during pollen development in tobacco

    Czech Academy of Sciences Publication Activity Database

    Hrubá, Petra; Honys, David; Tupý, Jaroslav

    2005-01-01

    Roč. 18, č. 3 (2005), s. 143-148 ISSN 0934-0882 R&D Projects: GA ČR GP522/02/D075; GA MŠk LZ1K03018; GA AV ČR KJB6038409 Institutional research plan: CEZ:AV0Z50380511 Keywords : tobacco * pollen development * thermotolerance Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 1.278, year: 2005

  6. Thermotolerant fermenting yeasts for simultaneous saccharification fermentation of lignocellulosic biomass

    Directory of Open Access Journals (Sweden)

    Jairam Choudhary

    2016-05-01

    Full Text Available Lignocellulosic biomass is the most abundant renewable source of energy that has been widely explored as second-generation biofuel feedstock. Despite more than four decades of research, the process of ethanol production from lignocellulosic (LC biomass remains economically unfeasible. This is due to the high cost of enzymes, end-product inhibition of enzymes, and the need for cost-intensive inputs associated with a separate hydrolysis and fermentation (SHF process. Thermotolerant yeast strains that can undergo fermentation at temperatures above 40°C are suitable alternatives for developing the simultaneous saccharification and fermentation (SSF process to overcome the limitations of SHF. This review describes the various approaches to screen and develop thermotolerant yeasts via genetic and metabolic engineering. The advantages and limitations of SSF at high temperatures are also discussed. A critical insight into the effect of high temperatures on yeast morphology and physiology is also included. This can improve our understanding of the development of thermotolerant yeast amenable to the SSF process to make LC ethanol production commercially viable.

  7. Cell wall anchoring of the Campylobacter antigens to Lactococcus lactis

    Directory of Open Access Journals (Sweden)

    Patrycja Anna Kobierecka

    2016-02-01

    Full Text Available Campylobacter jejuni is the most frequent cause of human food-borne gastroenteritis and chicken meat is the main source of infection. Recent studies showed that broiler chicken immunization against Campylobacter should be the most efficient way to lower the number of human infections by this pathogen. Induction of the mucosal immune system after oral antigen administration should provide protective immunity to chickens. In this work we tested the usefulness of Lactococcus lactis, the most extensively studied lactic acid bacterium, as a delivery vector for Campylobacter antigens. First we constructed hybrid protein – CjaA antigen presenting CjaD peptide epitopes on its surface. We showed that specific rabbit anti-rCjaAD serum reacted strongly with both CjaA and CjaD produced by a wild type Campylobacter jejuni strain. Next, rCjaAD and CjaA were fused to the C-terminus of the L. lactis YndF containing the LPTXG motif. The genes expressing these proteins were transcribed under control of the L. lactis Usp45 promoter and their products contain the Usp45 signal sequences. This strategy ensures a cell surface location of both analysed proteins, which was confirmed by immunofluorescence assay. In order to evaluate the impact of antigen location on vaccine prototype efficacy, a L. lactis strain producing cytoplasm-located rCjaAD was also generated. Animal experiments showed a decrease of Campylobacter cecal load in vaccinated birds as compared with the control group and showed that the L. lactis harboring the surface-exposed rCjaAD antigen afforded greater protection than the L. lactis producing cytoplasm-located rCjaAD. To the best of our knowledge, this is the first attempt to employ LAB (Lactic Acid Bacteria strains as a mucosal delivery vehicle for chicken immunization. Although the observed reduction of chicken colonization by Campylobacter resulting from vaccination was rather moderate, the experiments showed that LAB strains can be considered

  8. Detection of Campylobacter spp. in chilled and frozen broiler carcasses comparing immunoassay, PCR and real time PCR methods

    OpenAIRE

    Reis, Luciana Pimenta; Menezes, Liliane Denize Miranda; Lima, Graciela Kunrath; Santos, Ethiene Luiza de Souza; Dorneles, Elaine Maria Seles; Assis, Débora Cristina Sampaio de; Lage, Andrey Pereira; Cançado, Silvana de Vasconcelos; Figueiredo, Tadeu Chaves de

    2018-01-01

    ABSTRACT: In order to detect and identify Campylobacter spp. in broiler chicken carcasses, and to compare detection methods, 43 chilled and 43 frozen carcasses were collected and analyzed. Three methodologies were evaluated: an automated Enzyme Linked Fluorescent Assay (ELFA) VIDAS®30, Polymerase Chain Reaction (PCR) and real-time PCR. Only four chilled carcasses (4.6%) were considered positive for Campylobacter spp. by VIDAS®30 and no sample was positive when the conventional PCR technique w...

  9. Ganglioside GM1 mimicry in Campylobacter strains from sporadic infections in the United States.

    Science.gov (United States)

    Nachamkin, I; Ung, H; Moran, A P; Yoo, D; Prendergast, M M; Nicholson, M A; Sheikh, K; Ho, T; Asbury, A K; McKhann, G M; Griffin, J W

    1999-05-01

    To determine whether GM1-like epitopes in Campylobacter species are specific to O serotypes associated with Guillain-Barré syndrome (GBS) or whether they are frequent among random Campylobacter isolates causing enteritis, 275 random enteritis-associated isolates of Campylobacter jejuni were analyzed. To determine whether GM1-like epitopes in Campylobacter species are specific to O serotypes associated with Guillan-Barre syndrome (GBS) or whether they are frequent among random Campylobacter isolates causing enteritis, 275 enteritis-associated isolates, randomly collected in the United States, were analyzed using a cholera-toxin binding assay [corrected]. Overall, 26.2% of the isolates were positive for the GM1-like epitope. Of the 36 different O serotypes in the sample, 21 (58.3%) contained no strains positive for GM1, whereas in 6 serotypes (16.7%), >50% of isolates were positive for GM1. GBS-associated serotypes were more likely to contain strains positive for GM1 than were non-GBS-associated serotypes (37.8% vs. 15.1%, P=.0116). The results suggest that humans are frequently exposed to strains exhibiting GM1-like mimicry and, while certain serotypes may be more likely to possess GM1-like epitopes, the presence of GM1-like epitopes on Campylobacter strains does not itself trigger GBS.

  10. Campylobacter spp. and birds of prey.

    Science.gov (United States)

    Dipineto, Ludovico; De Luca Bossa, Luigi Maria; Russo, Tamara Pasqualina; Cutino, Eridania Annalisa; Gargiulo, Antonio; Ciccarelli, Francesca; Raia, Pasquale; Menna, Lucia Francesca; Fioretti, Alessandro

    2014-06-01

    A total of 170 birds of prey admitted to two Wildlife Rescue and Rehabilitation Centers of Italy were examined. Birds were divided by diurnal (n = 15) and nocturnal (n = 7) species, sampled by cloacal swabs, and examined for Campylobacter spp. by cultural and molecular methods. Campylobacter spp. were isolated in 43 out of the 170 (25.3%) birds of prey examined. Among these, 43/43 (100%) were identified as Campylobacter jejuni and 10/43 (23.3%) were identified as Campylobacter coli recovered from mixed infections. Diurnal birds of prey showed a significantly higher prevalence value (P = 0.0006) for Campylobacter spp. than did nocturnal birds of prey.

  11. Global Epidemiology of Campylobacter Infection

    Science.gov (United States)

    Kaakoush, Nadeem O.; Castaño-Rodríguez, Natalia; Mitchell, Hazel M.

    2015-01-01

    SUMMARY Campylobacter jejuni infection is one of the most widespread infectious diseases of the last century. The incidence and prevalence of campylobacteriosis have increased in both developed and developing countries over the last 10 years. The dramatic increase in North America, Europe, and Australia is alarming, and data from parts of Africa, Asia, and the Middle East indicate that campylobacteriosis is endemic in these areas, especially in children. In addition to C. jejuni, there is increasing recognition of the clinical importance of emerging Campylobacter species, including Campylobacter concisus and Campylobacter ureolyticus. Poultry is a major reservoir and source of transmission of campylobacteriosis to humans. Other risk factors include consumption of animal products and water, contact with animals, and international travel. Strategic implementation of multifaceted biocontrol measures to reduce the transmission of this group of pathogens is paramount for public health. Overall, campylobacteriosis is still one of the most important infectious diseases that is likely to challenge global health in the years to come. This review provides a comprehensive overview of the global epidemiology, transmission, and clinical relevance of Campylobacter infection. PMID:26062576

  12. Global Epidemiology of Campylobacter Infection.

    Science.gov (United States)

    Kaakoush, Nadeem O; Castaño-Rodríguez, Natalia; Mitchell, Hazel M; Man, Si Ming

    2015-07-01

    Campylobacter jejuni infection is one of the most widespread infectious diseases of the last century. The incidence and prevalence of campylobacteriosis have increased in both developed and developing countries over the last 10 years. The dramatic increase in North America, Europe, and Australia is alarming, and data from parts of Africa, Asia, and the Middle East indicate that campylobacteriosis is endemic in these areas, especially in children. In addition to C. jejuni, there is increasing recognition of the clinical importance of emerging Campylobacter species, including Campylobacter concisus and Campylobacter ureolyticus. Poultry is a major reservoir and source of transmission of campylobacteriosis to humans. Other risk factors include consumption of animal products and water, contact with animals, and international travel. Strategic implementation of multifaceted biocontrol measures to reduce the transmission of this group of pathogens is paramount for public health. Overall, campylobacteriosis is still one of the most important infectious diseases that is likely to challenge global health in the years to come. This review provides a comprehensive overview of the global epidemiology, transmission, and clinical relevance of Campylobacter infection. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  13. 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...

  14. Sensitization to hyperthermia (450C) of normal and thermotolerant CHO cells by anisotonic media

    International Nuclear Information System (INIS)

    Henle, K.J.; Dethlefsen, L.A.

    1979-01-01

    Asynchronous CHO cells heated in the presence of hypertonic medium (600 mosM) showed an increased sensitivity to 45 0 C hyperthermia and the extent varied as a function of the time of heating with respect to the hypertonic shock. Hypotonic medium (150 mosM) also reduced n without significantly reducing the D 0 , and in contrast to hypertonic medium, this effect was independent of the temporal relationship between the osmotic shock and heating. Sensitization of non-thermotolerant cells by hypotonic shock was possible but only when cells were heated immediately upon return to isotonic medium following a one-hour equilibration in hypotonic medium. Thermotolerant CHO cells were sensitized slightly more than non-thermotolerant cells (a factor of 2.2 versus 1.7) when heated immediately after the hypertonic shock, and heat conditioning in the presence of hypertonic medium also sensitized the cells but not as dramatically as when thermotolerant cells were reheated in the presence of hypertonic medium (thermotolerant control D 0 = 16.8 +- 1.3 min versus 11.4 +- 2.4 and 7.8 +- 0.2, respectively for the hypertonic studies). In contrast to the studies with non-thermotolerant cells, thermotolerant cells were sensitized when heated immediately after the hypotonic shock (D 0 = 10.7 +- 1.6 min). Also, incubating the heat-conditioned cells in hypertonic medium impaired the development of thermotolerance. These data suggest that: (1) anisotonic-media reduction of the extrapolation number is associated with osmotically induced membrane stress. (2) The difference in sensitization between thermotolerant and non-thermotolerant cells is apparently one of quantity and not quality, and (3) in contrast to bacterial data, these mammalian data do not suggest a possible role for the altered intracellular ion concentrations per se in the development of thermotolerance. (author)

  15. 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

  16. Antimicrobial resistance of thermophilic Campylobacter

    DEFF Research Database (Denmark)

    Aarestrup, Frank Møller; Engberg, J.

    2001-01-01

    Campylobacter has become the leading cause of zoonotic enteric infections in developed and developing countries world-wide. Antimicrobial resistance has emerged among Campylobacter mainly as a consequence of the use of antimicrobial agents in food animal production. Resistance to drugs of choice...... for the treatment of infections, macrolides and fluoroquinolones has emerged as a clinical problem and interventions to reduce this are recommended. Resistance to fluoroquinolones and macrolides is mediated by chromosomal mutations. Resistance to other relevant antimicrobial agents, mediated by acquired resistance...... genes, has not become widespread so far. However, resistance genes originating from both Gram-positive and Gram-negative bacterial species have been found, showing the potential for acquired resistance to emerge in Campylobacter....

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

    Directory of Open Access Journals (Sweden)

    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

  18. 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

  19. A comparative study of conventional and molecular techniques in diagnosis of campylobacter gastroenteritis in children.

    Science.gov (United States)

    Ghosh, Roumi; Uppal, Beena; Aggarwal, Prabhav; Chakravarti, Anita; Jha, Arun Kumar; Dubey, A P

    2014-01-01

    Campylobacter species are a significant cause of gastroenteritis among children worldwide. Conventional methods for detection of Campylobacter spp. based on cultural isolation and biochemical tests are cumbersome and time consuming. Because of their superior sensitivity and cost effectiveness, molecular methods are often used for identification of the pathogens. To evaluate different diagnostic methods for identification of Campylobacter. Faecal samples were collected from 585 children (age ≤ 12 years) with acute diarrhoea admitted in a tertiary-care hospital, excluding children already on antimicrobial therapy. All samples were examined by four methods: Grams' staining, culture methods, Enzyme-Immuno Assay, and Polymerase Chain Reaction (PCR). After Grams' staining, samples were inoculated on modified charcoal cefoperazone deoxycholate agar. ProSpecT™ Microplate Assay® and PCR assay using cadF gene was done for detection of Campylobacter specific antigen and DNA, respectively, in faecal samples. McNemar's test was used to compare the results wherever applicable. 197 cases (33.67%) were found to be positive for Campylobacter by at least one method. But only 121 (20.78%) out of the 585 stool specimens tested fulfilled the positivity criteria, i.e., positive either by culture or by any two tests among other three. Culture had very low sensitivity (37.19%), whereas PCR had the highest (96.69%) sensitivity but lowest positive predictive value (86.03%). Rapid Grams' staining technique (sensitivity 63.64%) was found to be better than culture. Detection by PCR and ELISA was significantly better than by culture on selective media and Grams' staining (pdetection rates of Campylobacter in children with diarrhoea. However, enzyme-immuno assay with high accuracy has the advantage of applicability in resource-poor settings.

  20. Detection of Campylobacter species and Arcobacter butzleri in stool samples by use of real-time multiplex PCR.

    Science.gov (United States)

    de Boer, Richard F; Ott, Alewijn; Güren, Pinar; van Zanten, Evert; van Belkum, Alex; Kooistra-Smid, Anna M D

    2013-01-01

    The presence of Campylobacter (or Campylobacter-like) species in stools from patients suspected of infectious gastroenteritis (n = 493) was investigated using real-time PCR for detection of Arcobacter butzleri (hsp60 gene), Campylobacter coli (ceuE gene), Campylobacter jejuni (mapA), five acknowledged pathogenic Campylobacter spp. (C16S_Lund assay), and the Campylobacter genus (C16S_LvI assay). In total, 71.4% of the samples were positive for Campylobacter DNA (n = 352) by a Campylobacter genus-specific (C16S_LvI) assay. A total of 23 samples (4.7%) were positive in the C16S_Lund assay, used for detection of C. jejuni, C. coli, C. lari, C. upsaliensis, and C. hyointestinalis. Subsequent identification of these samples yielded detection frequencies (DF) of 4.1% (C. jejuni), 0.4% (C. coli), and 0.4% (C. upsaliensis). The DF of A. butzleri was 0.4%. Interestingly, sequencing of a subgroup (n = 46) of C16S_LvI PCR-positive samples resulted in a considerable number of Campylobacter concisus-positive samples (n = 20). PCR-positive findings with the C16S_Lund and C. jejuni/C. coli-specific assays were associated with more serious clinical symptoms (diarrhea and blood). Threshold cycle (C(T)) values of C. jejuni/C. coli PCR-positive samples were comparable to those of the C16S_Lund PCR (P = 0.21). C(T) values for both assays were significantly lower than those of the C16S_LvI assay (P < 0.001 and P < 0.00001, respectively). In conclusion, this study demonstrated that in combination, the C. jejuni/C coli-specific assays and the C16S_Lund assay are both useful for routine screening purposes. Furthermore, the DF of the emerging pathogen C. concisus was at least similar to the DF of C. jejuni.

  1. Quantification of Campylobacter spp. in pig feces by direct real-time PCR with an internal control of extraction and amplification.

    Science.gov (United States)

    Leblanc-Maridor, Mily; Garénaux, Amélie; Beaudeau, François; Chidaine, Bérangère; Seegers, Henri; Denis, Martine; Belloc, Catherine

    2011-04-01

    The rapid and direct quantification of Campylobacter spp. in complex substrates like feces or environmental samples is crucial to facilitate epidemiological studies on Campylobacter in pig production systems. We developed a real-time PCR assay for detecting and quantifying Campylobacter spp. directly in pig feces with the use of an internal control. Campylobacter spp. and Yersinia ruckeri primers-probes sets were designed and checked for specificity with diverse Campylobacter, related organisms, and other bacterial pathogens before being used in field samples. The quantification of Campylobacter spp. by the real-time PCR then was realized on 531 fecal samples obtained from experimentally and naturally infected pigs; the numeration of Campylobacter on Karmali plate was done in parallel. Yersinia ruckeri, used as bacterial internal control, was added to the samples before DNA extraction to control DNA-extraction and PCR-amplification. The sensitivity of the PCR assay was 10 genome copies. The established Campylobacter real-time PCR assay showed a 7-log-wide linear dynamic range of quantification (R²=0.99) with a detection limit of 200 Colony Forming Units of Campylobacter per gram of feces. A high correlation was found between the results obtained by real-time PCR and those by culture at both qualitative and quantitative levels. Moreover, DNA extraction followed by real-time PCR reduced the time needed for analysis to a few hours (within a working day). In conclusion, the real-time PCR developed in this study provides new tools for further epidemiological surveys to investigate the carriage and excretion of Campylobacter by pigs. Copyright © 2011 Elsevier B.V. All rights reserved.

  2. Campylobacter bacteremia: A rare and under-reported event?

    NARCIS (Netherlands)

    Louwen, R.; Baarlen, van P.; Vliet, van A.H.M.; Belkum, van A.; Hays, J.P.; Endtz, H.P.

    2012-01-01

    Bacteria belonging to the species Campylobacter are the most common cause of bacterial diarrhoea in humans. The clinical phenotype associated with Campylobacter infections ranges from asymptomatic conditions to severe colitis and bacteremia. In susceptible patients, Campylobacter infections are

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

    Directory of Open Access Journals (Sweden)

    Ada Cean

    2013-10-01

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

  4. Resistance to β-lactam and tetracycline in Campylobacter spp.isolated from broiler slaughterhouses in southern Brazil

    Directory of Open Access Journals (Sweden)

    Yuli M. Sierra-Arguello

    2015-07-01

    Full Text Available Abstract The study was carried out to screen and analyze the genetic characteristics of antimicrobial resistance in Campylobacter spp. from poultry sources. A total of 141 strains of Campylobacter isolated from samples of broilers of slaughterhouses in southern Brazil was identified by phenotypic and genotypic methods. Campylobacter isolates were evaluated for its antimicrobial susceptibility and the presence of resistance genes. The strains were investigated for antimicrobial susceptibility against two agents (ampicillin and tetracycline by disk diffusion method. PCR assay was used to confirm the specie and the presence of ampicillin (blaOXA-61, tetracycline tet(O, and the energy-dependent multi-drug efflux pump (cmeB genes. Campylobacter jejuni was the most ubiquitous; its presence was determined in 140 samples out of 141 (99.3%, whereas Campylobacter coli was found only in one of the contaminated samples (0.70%. The results obtained showed 65% and 35.5% of Campylobacter isolates resistant to β-lactams and tetracyclines, respectively. The cmeB gene responsible for multidrug resistance was detected in 26 isolates out 141 strains (18.5%. Moreover, 36 out of 141 Campylobacter strains (25.6% were found to be resistant to at least two different antimicrobia resistance markers (β-lactams and tetracyclines.

  5. Acquired Thermotolerance and Heat Shock Proteins in Thermophiles from the Three Phylogenetic Domains

    DEFF Research Database (Denmark)

    Trent, Jonathan D.; Gabrielsen, Mette; Jensen, Bo

    1994-01-01

    Thermophilic organisms from each of the three phylogenetic domains (Bacteria, Archaea, and Eucarya) acquired thermotolerance after heat shock. Bacillus caldolyticus grown at 60 degrees C and heat shocked at 69 degrees C for 10 min showed thermotolerance at 74 degrees C, Sulfolobus shibatae grown...

  6. Photosynthetic thermotolerance of woody savanna species in China is correlated with leaf life span.

    NARCIS (Netherlands)

    Zhang, J.L.; Poorter, L.; Hao, G.Y.; Cao, K.F.

    2012-01-01

    Background and Aims Photosynthetic thermotolerance (PT) is important for plant survival in tropical and sub-tropical savannas. However, little is known about thermotolerance of tropical and sub-tropical wild plants and its association with leaf phenology and persistence. Longer-lived leaves of

  7. Complete genome sequence of the thermotolerant foodborne pathogen Salmonella enterica serovar Senftenberg ATCC 43845 and phylogenetic analysis of loci encoding thermotolerance

    Science.gov (United States)

    Introduction: Previous studies in Cronobacter sakazakii, Klebsiella spp., and Escherichia coli have identified a genomic island that confers thermotolerance to its hosts. This island has recently been identified in Salmonella enterica serovar Senfentenberg ATCC 43845, a historically important, heat ...

  8. Campylobacter hyoilei Alderton et al. 1995 and Campylobacter coli Veron and Chatelain 1973 are subjective synonyms

    DEFF Research Database (Denmark)

    Vandamme, P.; VanDoorn, L.J.; AlRashid, S.T.

    1997-01-01

    The taxonomic affiliation of Campylobacter hyoilei was reevaluated by examining a variety of phenotypic and genotypic criteria. Whole cell protein electrophoresis and a comparison of 66 phenotypic characters revealed that reference strains of C. hyoilei were indistinguishable from Campylobacter...

  9. 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

  10. Lectin typing of Campylobacter concisus

    DEFF Research Database (Denmark)

    Aabenhus, Rune Munck; Hynes, Sean O; Permin, Henrik

    2002-01-01

    A total of 44 clinical isolates and the type strain of the putative pathogen Campylobacter concisus were grouped based on their reactions with plant lectins. The optimized lectin typing system used C. concisus strains proteolytically pretreated and subsequently typed by using a panel of four...

  11. Campylobacter pylori. Diagnosis and treatment

    NARCIS (Netherlands)

    Tytgat, G. N.; Rauws, E. A.; de Koster, E. H.

    1989-01-01

    There is now a plethora of methods to diagnose colonization of the human stomach with Campylobacter pylori, such as microbiological culture on various media, identification of C. pylori in biopsies or biopsy smears using various stains or neurological methods, serological demonstration of specific

  12. Construction of a thermotolerant Saccharomyces cerevisiae strain for bioethanol production with reduced fermentation time and saccharifying enzyme dose.

    Science.gov (United States)

    Lim, Ji Sung; Jang, You Ri; Lim, Young Hoon; Kim, Keun

    2012-10-01

    A thermotolerant Saccharomyces cerevisiae mutant strain, TT6, was constructed after multi-parental hybridization of five mutant strains obtained by UV or NTG treatment of the original strain, S. cerevisiae KV1. When incubated at 40 degrees C in YPD broth, TT6 began to grow exponentially in 10 h, but KV1 did not show any noticeable growth even after 22 h. The thermotolerant growth of TT6 was confirmed by serial dilution assay at 42 degrees C; TT6 grew at a cell concentration (10(-5)) 10,000 times lower than that of KV1 (10(-1)). Whereas ethanol production from YP containing 23%; (w/v) glucose by KV1 decreased with increasing temperature from 30 degrees C to 36 degrees C, ethanol production by TT6 did not decrease at temperatures up to 37 degrees C. When TT6 was tested for ethanol production at 36 degrees C by simultaneous saccharification and fermentation (SSF) from 23% corn, 24 h of fermentation time or 50% of the glucoamylase dose was saved when compared with KV1 at 30 degrees C. The ethanol yield from corn by SSF with TT6 at 36 degrees C was 91.7% of the theoretical yield, whereas that of KV1 at 30 degrees C was 90.6%.

  13. 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.

  14. Isolation and Identification of Campylobacter spp. from Poultry and Poultry By-Products in Tunisia by Conventional Culture Method and Multiplex Real-Time PCR.

    Science.gov (United States)

    Jribi, Hela; Sellami, Hanen; Mariam, Siala; Smaoui, Salma; Ghorbel, Asma; Hachicha, Salma; Benejat, Lucie; Messadi-Akrout, Feriel; Mégraud, Francis; Gdoura, Radhouane

    2017-10-01

    Thermophilic Campylobacter spp. are one of the primary causes of bacterial human diarrhea. The consumption of poultry meats, by-products, or both is suspected to be a major cause of human campylobacteriosis. The aims of this study were to determine the prevalence of thermophilic Campylobacter spp. in fresh poultry meat and poultry by-products by conventional culture methods and to confirm Campylobacter jejuni and Campylobacter coli isolates by using the multiplex PCR assay. Two hundred fifty fresh poultry samples were collected from a variety of supermarkets and slaughterhouses located in Sfax, Tunisia, including chicken (n =149) and turkey (n =101). The samples were analyzed using conventional microbiological examinations according to the 2006 International Organization for Standardization method (ISO 10272-1) for Campylobacter spp. Concurrently, a real-time PCR was used for identification of C. jejuni and C. coli . Of the 250 samples of poultry meat and poultry by-products, 25.6% (n = 64) were contaminated with Campylobacter spp. The highest prevalence of Campylobacter spp. was found in chicken meat (26.8%) followed by turkey meat (23.7%). Among the different products, poultry breasts showed the highest contamination (36.6%) followed by poultry by-products (30%), poultry wings (28%) and poultry legs (26%) showed the lowest contamination, and no contamination was found on neck skin. Of the 64 thermophilic Campylobacter isolates, C. jejuni (59.7%) was the most frequently isolated species and 10.9% of the isolates were identified as C. coli . All of the 64 Campylobacter isolates identified by the conventional culture methods were further confirmed by PCR. The seasonal peak of Campylobacter spp. contamination was in the warm seasons (spring and summer). The study concluded that high proportions of poultry meat and poultry by-products marketed in Tunisia are contaminated by Campylobacter spp. Furthermore, to ensure food safety, poultry meats must be properly cooked

  15. Antimicrobial resistance of Campylobacter isolates from small scale and backyard chicken in Kenya.

    Science.gov (United States)

    Nguyen, Tuan Ngoc Minh; Hotzel, Helmut; Njeru, John; Mwituria, Joyce; El-Adawy, Hosny; Tomaso, Herbert; Neubauer, Heinrich; Hafez, Hafez M

    2016-01-01

    Thermophilic Campylobacter species are a major cause of bacterial foodborne diarrhoea in humans worldwide. Poultry and their products are the predominant source for human campylobacteriosis. Resistance of Campylobacter to antibiotics is increasing worldwide, but little is known about the antibiotic resistance in Campylobacter isolated from chicken in Kenya. In this study, 35 suspected Campylobacter strains isolated from faeces and cloacal swabs of chicken were tested for their susceptibility to seven antibiotics using a broth microdilution assay and molecular biological investigations. Overall, DNA of thermophilic Campylobacter was identified in 53 samples by PCR (34 C. jejuni, 18 C. coli and one mix of both species) but only 35 Campylobacter isolates (31 C. jejuni and 4 C. coli) could be re-cultivated after transportation to Germany. Isolates were tested for their susceptibility to antibiotics using a broth microdilution assay. Additionally, molecular biological detection of antibiotic resistance genes was carried out. C. jejuni isolates showed a high rate of resistance to nalidixic acid, tetracycline and ciprofloxacin of 77.4, 71.0 and 71.0 %, respectively. Low resistance (25.8 %) was detected for gentamicin and chloramphenicol. Multidrug resistance in C. jejuni could be detected in 19 (61.3 %) isolates. Resistance pattern of C. coli isolates was comparable. Resistance to ciprofloxacin was confirmed by MAMA-PCR and PCR-RFLP in all phenotypically resistant isolates. The tet(O) gene was detected only in 54.5 % of tetracycline resistant C. jejuni isolates. The tet(A) gene, which is also responsible for tetracycline resistance, was found in 90.3 % of C. jejuni and in all C. coli isolates. Thirteen phenotypically erythromycin-resistant isolates could not be characterised by using PCR-RFLP and MAMA-PCR. To the best of our knowledge, this study is the first report about resistance to antibiotics in thermophilic Campylobacter originating from chicken in Kenya

  16. 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

  17. Oxidative stress and antioxidant response in a thermotolerant yeast.

    Science.gov (United States)

    Mejía-Barajas, Jorge A; Montoya-Pérez, Rocío; Salgado-Garciglia, Rafael; Aguilera-Aguirre, Leopoldo; Cortés-Rojo, Christian; Mejía-Zepeda, Ricardo; Arellano-Plaza, Melchor; Saavedra-Molina, Alfredo

    Stress tolerance is a key attribute that must be considered when using yeast cells for industrial applications. High temperature is one factor that can cause stress in yeast. High environmental temperature in particular may exert a natural selection pressure to evolve yeasts into thermotolerant strains. In the present study, three yeasts (Saccharomyces cerevisiae, MC4, and Kluyveromyces marxianus, OFF1 and SLP1) isolated from hot environments were exposed to increased temperatures and were then compared with a laboratory yeast strain. Their resistance to high temperature, oxidative stress, and antioxidant response were evaluated, along with the fatty acid composition of their cell membranes. The SLP1 strain showed a higher specific growth rate, biomass yield, and biomass volumetric productivity while also showing lower duplication time, reactive oxygen species (ROS) production, and lipid peroxidation. In addition, the SLP1 strain demonstrated more catalase activity after temperature was increased, and this strain also showed membranes enriched in saturated fatty acids. It is concluded that the SLP1 yeast strain is a thermotolerant yeast with less oxidative stress and a greater antioxidant response. Therefore, this strain could be used for fermentation at high temperatures. Copyright © 2016 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.

  18. Oxidative stress and antioxidant response in a thermotolerant yeast

    Directory of Open Access Journals (Sweden)

    Jorge A. Mejía-Barajas

    Full Text Available Abstract Stress tolerance is a key attribute that must be considered when using yeast cells for industrial applications. High temperature is one factor that can cause stress in yeast. High environmental temperature in particular may exert a natural selection pressure to evolve yeasts into thermotolerant strains. In the present study, three yeasts (Saccharomyces cerevisiae, MC4, and Kluyveromyces marxianus, OFF1 and SLP1 isolated from hot environments were exposed to increased temperatures and were then compared with a laboratory yeast strain. Their resistance to high temperature, oxidative stress, and antioxidant response were evaluated, along with the fatty acid composition of their cell membranes. The SLP1 strain showed a higher specific growth rate, biomass yield, and biomass volumetric productivity while also showing lower duplication time, reactive oxygen species (ROS production, and lipid peroxidation. In addition, the SLP1 strain demonstrated more catalase activity after temperature was increased, and this strain also showed membranes enriched in saturated fatty acids. It is concluded that the SLP1 yeast strain is a thermotolerant yeast with less oxidative stress and a greater antioxidant response. Therefore, this strain could be used for fermentation at high temperatures.

  19. Effects of proliferation on the decay of thermotolerance in Chinese hamster cells.

    Science.gov (United States)

    Armour, E P; Li, G C; Hahn, G M

    1985-09-01

    Development and decay of thermotolerance were observed in Chinese hamster HA-1 cells. The thermotolerance kinetics of exponentially growing and fed plateau-phase cells were compared. Following a 10-min heat exposure at 45 degrees C, cells in both growth states had similar rates of development of tolerance to a subsequent 45-min exposure at 45 degrees C. This thermotolerant state started to decay between 12 and 24 hr after the initial heat exposure. The decay appeared to initiate slightly sooner in the exponentially growing cells when compared to the fed plateau-phase cells. During the decay phase, the rate of thermotolerance decay was similar in the two growth conditions. In other experiments, cells were induced to divide at a slower rate by chronic growth (3 months) in a low concentration of fetal calf serum. Under these low serum conditions cells became more sensitive to heat and the rate of decay of thermotolerance remained the same for exponentially growing cells. Plateau-phase cells were also more sensitive, but thermotolerance decayed more rapidly in these cells. Although dramatic cell cycle perturbations were seen in the exponentially growing cells, these changes appeared not to be related to thermotolerance kinetics.

  20. Genome Reduction for Niche Association in Campylobacter Hepaticus, A Cause of Spotty Liver Disease in Poultry

    Directory of Open Access Journals (Sweden)

    Liljana Petrovska

    2017-08-01

    Full Text Available The term “spotty liver disease” (SLD has been used since the late 1990s for a condition seen in the UK and Australia that primarily affects free range laying hens around peak lay, causing acute mortality and a fall in egg production. A novel thermophilic SLD-associated Campylobacter was reported in the United Kingdom (UK in 2015. Subsequently, similar isolates occurring in Australia were formally described as a new species, Campylobacter hepaticus. We describe the comparative genomics of 10 C. hepaticus isolates recovered from 5 geographically distinct poultry holdings in the UK between 2010 and 2012. Hierarchical gene-by-gene analyses of the study isolates and representatives of 24 known Campylobacter species indicated that C. hepaticus is most closely related to the major pathogens Campylobacter jejuni and Campylobacter coli. We observed low levels of within-farm variation, even between isolates collected over almost 3 years. With respect to C. hepaticus genome features, we noted that the study isolates had a ~140 Kb reduction in genome size, ~144 fewer genes, and a lower GC content compared to C. jejuni. The most notable reduction was in the subsystem containing genes for iron acquisition and metabolism, supported by reduced growth of C. hepaticus in an iron depletion assay. Genome reduction is common among many pathogens and in C. hepaticus has likely been driven at least in part by specialization following the occupation of a new niche, the chicken liver.

  1. Detection of Campylobacter from poultry carcass skin samples at slaughter in Southern Italy.

    Science.gov (United States)

    Pepe, Tiziana; De Dominicis, Rosaria; Esposito, Giuseppina; Ventrone, Iole; Fratamico, Pina M; Cortesi, Maria Luisa

    2009-08-01

    Campylobacter is a major foodborne pathogen responsible for acute gastroenteritis characterized by diarrhea that is sometimes bloody, fever, cramps, and vomiting. Campylobacter species are carried in the intestinal tracts of mammals and birds, and sources of human infection include raw milk, contaminated water, direct contact with pets, and foods, particularly poultry. Campylobacter jejuni and C. coli are the species that account for the majority of human infections. The aim of this work was to determine the prevalence of Campylobacter in 190 poultry carcasses sampled at slaughter and to use a multiplex PCR assay to determine if the isolates were C. jejuni or C. coli. C. coli was not isolated, while C. jejuni was recovered from 52 (37.1%) of 140 carcasses for which pools of four sampling sites (neck, cloaca, breast, and back) were examined. In the remaining 50 carcasses, the four sites were analyzed separately, and C. jejuni was recovered from the samples in the following order: neck (n = 20), cloaca (n = 16), breast (n = 14), and back (n = 11). The results are in agreement with those of other studies, which showed that C. jejuni is more commonly associated with poultry than is C. coli. Control strategies for Campylobacter should include interventions to eliminate C. jejuni in poultry at various stages of production and processing, including at slaughter.

  2. The structural-functional organization of thermotolerant complexes of actinomycetes in desert and volcanic soils

    Science.gov (United States)

    Zenova, G. M.; Kurapova, A. I.; Lysenko, A. M.; Zvyagintsev, D. G.

    2009-05-01

    It has been found that the number of thermotolerant actinomycetes in strongly heated soils of deserts and volcanic regions is comparable to or exceeds the number of mesophilic actinomycetes. Among the latter group, streptomyces usually predominate; among thermotolerant actinomycetes, representatives of the Micromonospora, Streptosporangium, Actinomadura, Saccharopolyspora, Microtetraspora, and Microbispora genera are identified. Thermotolerant actinomycetes display the full cycle of their development in these soils. The method of fluorescent in situ hybridization has made it possible to determine that mycelial forms predominate among the metabolically active representatives of Actinobacteria; their portion increases with the rise in the temperature of soil incubation.

  3. Thermotolerance-induced goblet cell activity confers protection in post-operative gut barrier dysfunction.

    LENUS (Irish Health Repository)

    Ali, Rohana

    2009-06-01

    There is evidence that some level of protection against the adverse sequelae of surgery is provided by induction of thermotolerance; this protective effect was explored by study of several indicators of bowel wall damage in animals exposed to surgical insults. It has been argued that the mechanism of the protective effect of thermotolerance involves heat shock proteins (HSPs). We hypothesized that the protective effect of thermotolerance may be due in part to changes in the bowel wall itself, and we investigated this hypothesis in an experimental rat model.

  4. Campylobacter virulence and survival factors.

    Science.gov (United States)

    Bolton, Declan J

    2015-06-01

    Despite over 30 years of research, campylobacteriosis is the most prevalent foodborne bacterial infection in many countries including in the European Union and the United States of America. However, relatively little is known about the virulence factors in Campylobacter or how an apparently fragile organism can survive in the food chain, often with enhanced pathogenicity. This review collates information on the virulence and survival determinants including motility, chemotaxis, adhesion, invasion, multidrug resistance, bile resistance and stress response factors. It discusses their function in transition through the food processing environment and human infection. In doing so it provides a fundamental understanding of Campylobacter, critical for improved diagnosis, surveillance and control. Copyright © 2014 Elsevier Ltd. All rights reserved.

  5. Lectin typing of Campylobacter concisus

    DEFF Research Database (Denmark)

    Aabenhus, Rune Munck; Hynes, Sean O; Permin, Henrik

    2002-01-01

    A total of 44 clinical isolates and the type strain of the putative pathogen Campylobacter concisus were grouped based on their reactions with plant lectins. The optimized lectin typing system used C. concisus strains proteolytically pretreated and subsequently typed by using a panel of four...... lectins. The system grouped all 45 strains into 13 lectin reaction patterns, leaving no strain untypeable due to autoagglutination. Lectin types were both stable and reproducible....

  6. Neutrophil activation by Campylobacter concisus

    OpenAIRE

    S?rensen, Nina B; Nielsen, Hans L; Varming, Kim; Nielsen, Henrik

    2013-01-01

    Background Campylobacter concisus is an emerging enteric pathogen associated with prolonged diarrhoea and possibly inflammatory bowel disease in children as well as adults, but the interaction with cells of the innate immune system is unclear. The magnitude of systemic immunoglobulin response in acute infection is unknown. Methods Neutrophils from healthy volunteers were activated with five faecal isolates of C. concisus from patients with gastroenteritis as well as the oral reference strain ...

  7. Prevalence of Thermophilic Campylobacter species in carcasses ...

    African Journals Online (AJOL)

    Background: Thermophilic Campylobacter spp. namely, Campylobacter jejuni and coli cause acute diarrheal diseases in humans worldwide; although these species are known to occur in the intestinal tract of a wide variety of domestic and wild animals. Objective: Little is known about the presence of these bacteria in ...

  8. 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

  9. Integrated approach leads to less Campylobacter

    DEFF Research Database (Denmark)

    Rosenquist, Hanne; Hald, Birthe; Borck Høg, Birgitte

    2012-01-01

    Methods of reducing the risk of Campylobacter infection during indoor broiler (chicken) production are discussed, including: risk management intiatives; biosecurity measures; scheduled slaughter; hygiene and decontamination; and improving consumer information.......Methods of reducing the risk of Campylobacter infection during indoor broiler (chicken) production are discussed, including: risk management intiatives; biosecurity measures; scheduled slaughter; hygiene and decontamination; and improving consumer information....

  10. Quantifying transmission of Campylobacter spp. among broilers

    NARCIS (Netherlands)

    Gerwe, van T.J.; Bouma, A.; Jacobs-Reitsma, W.F.; Broek, van den E.W.F.; Klinkenberg, D.; Stegeman, J.A.; Heesterbeek, J.A.P.

    2005-01-01

    Campylobacter species are frequently identified as a cause of human gastroenteritis, often from eating or mishandling contaminated poultry products. Quantitative knowledge of transmission of Campylobacter in broiler flocks is necessary, as this may help to determine the moment of introduction of

  11. Campylobacter prevalence in retail chicken liver

    Science.gov (United States)

    Foodborne campylobacteriosis has been linked to undercooked chicken liver. It is unknown how commonly chicken livers are contaminated with Campylobacter. The objective of this study was to determine the prevalence of Campylobacter on chicken livers available at retail. For each of five weeks, t...

  12. Flies and Campylobacter infection of broiler flocks

    DEFF Research Database (Denmark)

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

    2004-01-01

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

  13. 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

  14. 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.

  15. Campylobacter in poultry, pork and beef

    DEFF Research Database (Denmark)

    Josefsen, Mathilde Hasseldam; Carroll, C.; Rudi, K.

    2011-01-01

    Campylobacter infection has become one of the most important zoonoses worldwide. A low prevalence of Campylobacter is generally found in beef and pork at retail, although they may still be sources of infection. Based on the high prevalence of poultry-associated infections, this chapter mainly...... focuses on rapid methods for detection of Campylobacter in this particular production chain, and describes the routes of transmission and sampling in the different levels as well as intervention strategies. The chapter focuses on the introduction, infection dynamics, and sampling of Campylobacter...... throughout the poultry production chain, from farm to consumer level. It also describes culture-based, immunological, and molecular methods for rapid detection, characterization, and enumeration for Campylobacter. Rapid methods can generally be also more sensitive and specific than culture-based methods...

  16. 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.

  17. 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....

  18. The first closed genome sequence of Campylobacter fetus subsp. venerealis biovar intermedius

    Science.gov (United States)

    Campylobacter fetus venerealis biovar intermedius is a variant of Campylobacter fetus subsp. venerealis, the causative agent of Bovine Genital Campylobacteriosis. In contrast to Campylobacter fetus subsp. venerealis which is restricted to the genital tract of cattle, Campylobacter fetus subsp. vener...

  19. Detection of Campylobacter spp. in chicken fecal samples by real-time PCR

    DEFF Research Database (Denmark)

    Lund, Marianne; Nordentoft, Steen; Pedersen, Karl

    2004-01-01

    A real-time PCR assay for detecting thermophilic Campylobacter spp. directly in chicken feces has been developed. DNA was isolated from fecal material by using magnetic beads followed by PCR with a prealiquoted PCR mixture, which had been stored at -18degreesC. Campylobacter could be detected...... in less than 4 h, with a detection limit of 100 to 150 CFU/ml, in a fecal suspension. A bacterial internal control was added before DNA extraction to control both DNA isolation and the presence of PCR inhibitors in the samples. The assay was performed on 111 swab samples from a Danish surveillance program...... and compared to conventional culturing using selective enrichment. There was no statistically significant difference in performance between real-time PCR and culture by selective enrichment, and the diagnostic specificity was 0.96 with an agreement of 0.92. Therefore, the assay should be useful for screening...

  20. Prevalence and Antimicrobial Resistance of Campylobacter Isolated from Dressed Beef Carcasses and Raw Milk in Tanzania

    Science.gov (United States)

    Kashoma, Isaac P.; Kassem, Issmat I.; John, Julius; Kessy, Beda M.; Gebreyes, Wondwossen; Kazwala, Rudovick R.

    2016-01-01

    Campylobacter species are commonly transmitted to humans through consumption of contaminated foods such as milk and meat. The aim of this study was to investigate the prevalence, antimicrobial resistance, and genetic determinants of resistance of Campylobacter isolated from raw milk and beef carcasses in Tanzania. The antimicrobial resistance genes tested included blaOXA-61 (ampicillin), aph-3-1 (aminoglycoside), tet(O) (tetracycline), and cmeB (multi-drug efflux pump). The prevalence of Campylobacter was 9.5% in beef carcasses and 13.4% in raw milk, respectively. Using multiplex-polymerase chain reaction (PCR), we identified 58.1% of the isolates as Campylobacter jejuni, 30.7% as Campylobacter coli, and 9.7% as other Campylobacter spp. One isolate (1.6%) was positive for both C. jejuni and C. coli specific PCR. Antimicrobial susceptibility testing using the disk diffusion assay and the broth microdilution method showed resistance to: ampicillin (63% and 94.1%), ciprofloxacin (9.3% and 11.8%), erythromycin (53.7% and 70.6%), gentamicin (0% and 15.7%), streptomycin (35.2% and 84.3%), and tetracycline (18.5% and 17.7%), respectively. Resistance to azithromycin (42.6%), nalidixic acid (64.8%), and chloramphenicol (13%) was determined using the disk diffusion assay only, while resistance to tylosin (90.2%) was quantified using the broth microdilution method. The blaOXA-61 (52.6% and 28.1%), cmeB (26.3% and 31.3%), tet(O) (26.3% and 31.3%), and aph-3-1 (5.3% and 3.0%) were detected in C. coli and C. jejuni. These findings highlight the extent of antimicrobial resistance in Campylobacter occurring in important foods in Tanzania. The potential risks to consumers emphasize the need for adequate control approaches, including the prudent use of antimicrobials to minimize the spread of antimicrobial-resistant Campylobacter. PMID:26153978

  1. 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

  2. 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.

  3. 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.

  4. Thermotolerance and protein glycosylation: Inhibition studies with sodium fluoride, azauridine and tunicamycin

    International Nuclear Information System (INIS)

    Bursey, D.L.; Henle, K.J.; Nagle, W.A.; Moss, A.J.

    1987-01-01

    The glycosylation hypothesis predicts increased incorporation of monosaccharides into 0-linked glycoproteins during thermotolerance development and inhibition of thermotolerance when this process is blocked. Specific inhibitors of 0-linked glycosylation are not available. The authors examined the effect of non-specific inhibition of glycosylation on thermotolerance development by: 1. restriction of both exogenous sugars and endogeneous sugar synthesis with NaF to block glycolysis while providing L-glutamine as a substrate for ATP synthesis in the TCA cycle; or 2. inhibition of UDP-sugar synthesis using azauridine and tunicamycin. Inhibitors were added to cell cultures after heat conditioning (10 min, 45 0 ) and removed after 6 hr prior to 45 0 -test heating. Sugar deprivation was achieved with 10mM NaF in glucose-free EBSS, supplemented with 2mM L-glutamine. Synthesis of UDP-sugars was inhibited with 1mM azauridine + 1μg/ml tunicamycin. Thermotolerance development was inhibited 87% by NaF/glutamine and 47% by azauridine/tunicamycin. For example, the D/sub o/ of the thermotolerant cells was 42.5 min (control D/sub o/ = 3 min), but only 5.5 min with inhibition by the NaF solution. These results support the absolute requirement of sugar precursors for thermotolerance development as predicted by the glycosylation hypothesis

  5. Production of thermotolerant entomopathogenic Isaria fumosorosea SFP-198 conidia in corn-corn oil mixture.

    Science.gov (United States)

    Kim, Jae Su; Je, Yeon Ho; Roh, Jong Yul

    2010-04-01

    Low thermotolerance of entomopathogenic fungi is a major impediment to long-term storage and effective application of these biopesticides under seasonal high temperatures. The effects of high temperatures on the viability of an entomopathogenic fungus, Isaria fumosorosea SFP-198 (KCTC 0499BP), produced on different substrates amended with various additives were explored. Ground corn was found to be superior in producing the most thermotolerant conidia compared to yellow soybean, red kidney bean, and rice in a polyethylene bag production system. Using ground corn mixed with corn oil as a substrate resulted in only 7% reduction in germination compared to ground corn alone (67% reduction) after exposure of conidia to 50 degrees C for 2 h. Corn oil as an additive for ground corn was followed by inorganic salts (KCl and NaCl), carbohydrates (sucrose and dextrin), a sugar alcohol (sorbitol), and plant oils (soybean oil and cotton seed oil) in ability to improve conidial thermotolerance. Unsaturated fatty acids, such as linoleic acid and oleic acid, the main components of corn oil, served as effective additives for conidial thermotolerance in a dosage-dependent manner, possibly explaining the improvement by corn oil. This finding suggests that the corn-corn oil mixture can be used to produce highly thermotolerant SFP-198 conidia and provides the relation of unsaturated fatty acids as substrates with conidial thermotolerance.

  6. 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...

  7. Prevalence and antibiotic resistant of Campylobacter spp. isolated from different stages of sheep slaughterhouse

    Directory of Open Access Journals (Sweden)

    A Shakerian

    2012-02-01

    Full Text Available Campylobacter jejuni/coli are frequent causes of diarrhea in humans worldwide originating in foods of animal origin mainly from meat. The aim of this study was to determine the prevalence of Campylobacter spp. in lamb at different stages of the slaughter line including: after-skinning, after evisceration and the end of slaughter process. A total of 150 lamb samples (50 samples per each stage were collected over a period of 16-month between January 2006 and May 2008, and were analyzed for the presence of Campylobacter spp. According to the results, Campylobacter spp. were isolated from 11.3% (17/150 of the carcasses from the three sampling stages. Among the isolates, 76.5% were identified as C. jejuni and 23.1% as C. coli. Campylobacter spp. were isolated from 5%, 8% and 4% of carcasses during the stages of after-skinning, after-evisceration and the end of slaughter process, respectively. Antibiotics susceptibility of 17 isolates were determined for ten different antibiotics using the disk diffusion assay. Results revealed that 58/8% of the isolates were resistant to ciprofloxacin, while 47/1% of the isolates to nalidixic acid, 41/2% to tetracycline, 29/4% to enrofloxacin, 23/5% to ampicillin, 5/9% to amoxicillin, and 5/9% top streptomycine. None of the isolates was resistant to erythromycin, chloramphenicol and gentamicine. This study emphasizes the application of a preventive system such as HACCP (Hazard Analysis of Critical Control Points for the control of Campylobacter contamination in slaughterhouse.

  8. Antibacterial Activity of Glutathione-Stabilized Silver Nanoparticles Against Campylobacter Multidrug-Resistant Strains

    Directory of Open Access Journals (Sweden)

    Jose M. Silvan

    2018-03-01

    Full Text Available Campylobacter is the leading cause of bacterial diarrheal disease worldwide. Although most episodes of campylobacteriosis are self-limiting, antibiotic treatment is usually needed in patients with serious enteritis, and especially in childrens or the elderly. In the last years, antibiotic resistance in Campylobacter has become a major public health concern and a great interest exists in developing new antimicrobial strategies for reducing the impact of this food-borne pathogen on human health. Among them, the use of silver nanoparticles as antibacterial agents has taken on increased importance in the field of medicine. The aim of the present study was to evaluate the antimicrobial effectiveness of glutathione-stabilized silver nanoparticles (GSH-Ag NPs against multidrug resistant (MDR Campylobacter strains isolated from the chicken food chain (FC and clinical patients (C. The results obtained showed that GSH-Ag NPs were highly effective against all MDR Campylobacter strains tested. The minimal inhibitory concentration (MIC and minimal bactericidal concentration (MBC were in a range from 4.92 to 39.4 μg/mL and 9.85 to 39.4 μg/mL, respectively. Cytotoxicity assays were also assessed using human intestinal HT-29, Caco-2, and CCD-18 epithelial cells. Exposure of GSH-Ag NPs to intestinal cells showed a dose-dependent cytotoxic effect in all cell lines between 9.85 and 39.4 μg/mL. More than 60% of the tested Campylobacter strains were susceptible to GSH-Ag NPs concentrations ≤ 9.85 μg/mL, suggesting that practical inhibitory levels could be reached at low GSH-Ag NPs concentrations. Further works are needed with the purpose to evaluate the practical implications of the toxicity studies and to know more about other attributes linked to the biological compatibility. This behavior makes GSH-Ag NPs as a promising tool for the design of novel antibacterial agents for controlling Campylobacter.

  9. Molecular detection of Campylobacter spp. and fecal indicator bacteria during the northern migration of sandhill cranes (Grus canadensis) at the central Platte River.

    Science.gov (United States)

    Lu, Jingrang; Ryu, Hodon; Vogel, Jason; Santo Domingo, Jorge; Ashbolt, Nicholas J

    2013-06-01

    The risk to human health of the annual sandhill crane (Grus canadensis) migration through Nebraska, which is thought to be a major source of fecal pollution of the central Platte River, is unknown. To better understand potential risks, the presence of Campylobacter species and three fecal indicator bacterial groups (Enterococcus spp., Escherichia coli, and Bacteroidetes) was assayed by PCR from crane excreta and water samples collected during their stopover at the Platte River, Nebraska, in 2010. Genus-specific PCR assays and sequence analyses identified Campylobacter jejuni as the predominant Campylobacter species in sandhill crane excreta. Campylobacter spp. were detected in 48% of crane excreta, 24% of water samples, and 11% of sediment samples. The estimated densities of Enterococcus spp. were highest in excreta samples (mean, 4.6 × 10(8) cell equivalents [CE]/g), while water samples contained higher levels of Bacteroidetes (mean, 5.1 × 10(5) CE/100 ml). Enterococcus spp., E. coli, and Campylobacter spp. were significantly increased in river water and sediments during the crane migration period, with Enterococcus sp. densities (~3.3 × 10(5) CE/g) 2 to 4 orders of magnitude higher than those of Bacteroidetes (4.9 × 10(3) CE/g), E. coli (2.2 × 10(3) CE/g), and Campylobacter spp. (37 CE/g). Sequencing data for the 16S rRNA gene and Campylobacter species-specific PCR assays indicated that C. jejuni was the major Campylobacter species present in water, sediments, and crane excreta. Overall, migration appeared to result in a significant, but temporary, change in water quality in spring, when there may be a C. jejuni health hazard associated with water and crops visited by the migrating birds.

  10. 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...

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

    Science.gov (United States)

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

    2015-03-01

    During sampling of reptiles for members of the class Epsilonproteobacteria, strains representing a member of the genus Campylobacter not belonging to any of the established taxa were isolated from lizards and chelonians. Initial amplified fragment length polymorphism, PCR and 16S rRNA sequence analysis showed that these strains were most closely related to Campylobacter fetus and Campylobacter hyointestinalis. A polyphasic study was undertaken to determine the taxonomic position of five strains. The strains were characterized by 16S rRNA and atpA sequence analysis, matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry and conventional phenotypic testing. Whole-genome sequences were determined for strains 1485E(T) and 2463D, and the average nucleotide and amino acid identities were determined for these strains. The strains formed a robust phylogenetic clade, divergent from all other species of the genus Campylobacter. In contrast to most currently known members of the genus Campylobacter, the strains showed growth at ambient temperatures, which might be an adaptation to their reptilian hosts. The results of this study clearly show that these strains isolated from reptiles represent a novel species within the genus Campylobacter, for which the name Campylobacter iguaniorum sp. nov. is proposed. The type strain is 1485E(T) ( = LMG 28143(T) = CCUG 66346(T)). © 2015 IUMS.

  12. 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.

  13. 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.

  14. Campylobacter ureolyticus: an emerging gastrointestinal pathogen?

    LENUS (Irish Health Repository)

    Bullman, Susan

    2011-03-01

    A total of 7194 faecal samples collected over a 1-year period from patients presenting with diarrhoea were screened for Campylobacter spp. using EntericBio(®) , a multiplex-PCR system. Of 349 Campylobacter-positive samples, 23.8% were shown to be Campylobacter ureolyticus, using a combination of 16S rRNA gene analysis and highly specific primers targeting the HSP60 gene of this organism. This is, to the best of our knowledge, the first report of C. ureolyticus in the faeces of patients presenting with gastroenteritis and may suggest a role for this organism as an emerging enteric pathogen.

  15. Campylobacter in poultry, pork and beef

    DEFF Research Database (Denmark)

    Josefsen, Mathilde Hasseldam; Carroll, C.; Rudi, K.

    2011-01-01

    focuses on rapid methods for detection of Campylobacter in this particular production chain, and describes the routes of transmission and sampling in the different levels as well as intervention strategies. The chapter focuses on the introduction, infection dynamics, and sampling of Campylobacter...... throughout the poultry production chain, from farm to consumer level. It also describes culture-based, immunological, and molecular methods for rapid detection, characterization, and enumeration for Campylobacter. Rapid methods can generally be also more sensitive and specific than culture-based methods...

  16. Thermotolerant Yeasts for Bioethanol Production Using Lignocellulosic Substrates

    Science.gov (United States)

    Pasha, Chand; Rao, L. Venkateswar

    glucose without a physical and chemical pre-treatment. The pre-treatment processes normally applied on the different substrates are acidic hydrolysis, steam explosion and wet oxidation. A problem for most pretreatment methods is the generation of compounds that are inhibitory towards the fermenting microorganisms, primarily phenols. Degradation products that could have inhibitory action in later fermentation steps are avoided during pre-treatment by wet oxidation. Followed by pre treatment, hydrolysed with enzymes known as cellulases and hemicellulases, which hydrolyse cellulose and hemicellulose respectively. The production of bioethanol requires two steps, fermentation and distillation. Practically all ethanol fermentation is still based on Saccharomyces cerevisiae . The fermentation using thermotolerant yeasts has more advantageous in that they have faster fermentation rates, avoid the cooling costs, and decrease the over all fermentation costs, so that ethanol can be made available at cheaper rates. In addition they can be used for efficient simultaneous saccharification and fermentation of cellulose by cellulases because the temperature optimum of cellulase enzymes (about 40 ° C to 45 ° C) is close to the fermentation temperature of thermotolerant yeasts. Hence selection and improvement of thermotolerant yeasts for bioconversion of lignocellulosic substrates is very useful.

  17. Epidemiology and Impact of Campylobacter Infection in Children in 8 Low-Resource Settings: Results From the MAL-ED Study.

    Science.gov (United States)

    Amour, Caroline; Gratz, Jean; Mduma, Estomih; Svensen, Erling; Rogawski, Elizabeth T; McGrath, Monica; Seidman, Jessica C; McCormick, Benjamin J J; Shrestha, Sanjaya; Samie, Amidou; Mahfuz, Mustafa; Qureshi, Shahida; Hotwani, Aneeta; Babji, Sudhir; Trigoso, Dixner Rengifo; Lima, Aldo A M; Bodhidatta, Ladaporn; Bessong, Pascal; Ahmed, Tahmeed; Shakoor, Sadia; Kang, Gagandeep; Kosek, Margaret; Guerrant, Richard L; Lang, Dennis; Gottlieb, Michael; Houpt, Eric R; Platts-Mills, James A

    2016-11-01

     Enteropathogen infections have been associated with enteric dysfunction and impaired growth in children in low-resource settings. In a multisite birth cohort study (MAL-ED), we describe the epidemiology and impact of Campylobacter infection in the first 2 years of life.  Children were actively followed up until 24 months of age. Diarrheal and nondiarrheal stool samples were collected and tested by enzyme immunoassay for Campylobacter Stool and blood samples were assayed for markers of intestinal permeability and inflammation.  A total of 1892 children had 7601 diarrheal and 26 267 nondiarrheal stool samples tested for Campylobacter We describe a high prevalence of infection, with most children (n = 1606; 84.9%) having a Campylobacter-positive stool sample by 1 year of age. Factors associated with a reduced risk of Campylobacter detection included exclusive breastfeeding (risk ratio, 0.57; 95% confidence interval, .47-.67), treatment of drinking water (0.76; 0.70-0.83), access to an improved latrine (0.89; 0.82-0.97), and recent macrolide antibiotic use (0.68; 0.63-0.74). A high Campylobacter burden was associated with a lower length-for-age Z score at 24 months (-1.82; 95% confidence interval, -1.94 to -1.70) compared with a low burden (-1.49; -1.60 to -1.38). This association was robust to confounders and consistent across sites. Campylobacter infection was also associated with increased intestinal permeability and intestinal and systemic inflammation.  Campylobacter was prevalent across diverse settings and associated with growth shortfalls. Promotion of exclusive breastfeeding, drinking water treatment, improved latrines, and targeted antibiotic treatment may reduce the burden of Campylobacter infection and improve growth in children in these settings. © The Author 2016. Published by Oxford University Press for the Infectious Diseases Society of America.

  18. Effect of incubation temperature on the detection of thermophilic campylobacter species from freshwater beaches, nearby wastewater effluents, and bird fecal droppings.

    Science.gov (United States)

    Khan, Izhar U H; Hill, Stephen; Nowak, Eva; Edge, Thomas A

    2013-12-01

    This large-scale study compared incubation temperatures (37°C versus 42°C) to study the detection of thermophilic Campylobacter species, including Campylobacter jejuni, C. coli, and C. lari, in various surface water samples and bird fecal droppings around Hamilton Harbor, Lake Ontario. The putative culture isolates obtained from incubation temperatures of 37 and 42°C were confirmed by Campylobacter genus- and species-specific triplex PCR assays targeting the 16S rRNA gene and the 16S-23S rRNA gene internal transcribed spacer (ITS) region. A total of 759 water, wastewater, and bird fecal dropping samples were tested. Positive amplification reactions for the genus Campylobacter were found for 454 (60%) samples incubated at 37°C, compared to 258 (34%) samples incubated at 42°C. C. jejuni (16%) and C. lari (12%) were detected significantly more frequently at the 42°C incubation temperature than at 37°C (8% and 5%, respectively). In contrast, significantly higher rates of C. coli (14%) and other Campylobacter spp. (36%) were detected at the 37°C incubation temperature than at 42°C (8% and 7%, respectively). These results were consistent across surface water, wastewater, and bird fecal dropping samples. At times, Campylobacter spp. were recovered and detected at 37°C (3% for C. jejuni, 10% for C. coli, and 3% for C. lari) when the same samples incubated at 42°C were negative. A significantly higher rate of other Campylobacter spp. was detected only at 37°C (32%) than only at 42°C (3%). These results indicate that incubation temperature can significantly influence the culturability and detection of thermophilic and other fastidious Campylobacter spp. and that a comprehensive characterization of the Campylobacter spp. in surface water, wastewaters, or bird fecal droppings will require incubation at both 37 and 42°C.

  19. Detection and genotyping of Arcobacter and Campylobacter isolates from retail chicken samples by use of DNA oligonucleotide arrays.

    Science.gov (United States)

    Quiñones, Beatriz; Parker, Craig T; Janda, John M; Miller, William G; Mandrell, Robert E

    2007-06-01

    To explore the use of DNA microarrays for pathogen detection in food, we produced DNA oligonucleotide arrays to simultaneously determine the presence of Arcobacter and the presence of Campylobacter in retail chicken samples. Probes were selected that target housekeeping and virulence-associated genes in both Arcobacter butzleri and thermotolerant Campylobacter jejuni and Campylobacter coli. These microarrays showed a high level of probe specificity; the signal intensities detected for A. butzleri, C. coli, or C. jejuni probes were at least 10-fold higher than the background levels. Specific identification of A. butzleri, C. coli, and C. jejuni was achieved without the need for a PCR amplification step. By adapting an isolation method that employed membrane filtration and selective media, C. jejuni isolates were recovered from package liquid from whole chicken carcasses prior to enrichment. Increasing the time of enrichment resulted in the isolation of A. butzleri and increased the recovery of C. jejuni. C. jejuni isolates were further classified by using an additional subset of probes targeting the lipooligosaccharide (LOS) biosynthesis locus. Our results demonstrated that most of the C. jejuni isolates likely possess class B, C, or H LOS. Validation experiments demonstrated that the DNA microarray had a detection sensitivity threshold of approximately 10,000 C. jejuni cells. Interestingly, the use of C. jejuni sequence-specific primers to label genomic DNA improved the sensitivity of this DNA microarray for detection of C. jejuni in whole chicken carcass samples. C. jejuni was efficiently detected directly both in package liquid from whole chicken carcasses and in enrichment broths.

  20. The HaDREB2 transcription factor enhances basal thermotolerance and longevity of seeds through functional interaction with HaHSFA9.

    Science.gov (United States)

    Almoguera, Concepción; Prieto-Dapena, Pilar; Díaz-Martín, Juan; Espinosa, José M; Carranco, Raúl; Jordano, Juan

    2009-06-19

    Transcription factor HaDREB2 was identified in sunflower (Helianthus annuus L.) as a drought-responsive element-binding factor 2 (DREB2) with unique properties. HaDREB2 and the sunflower Heat Shock Factor A9 (HaHSFA9) co-activated the Hahsp17.6G1 promoter in sunflower embryos. Both factors could be involved in transcriptional co-activation of additional small heat stress protein (sHSP) promoters, and thus contribute to the HaHSFA9-mediated enhancement of longevity and basal thermotolerance of seeds. We found that overexpression of HaDREB2 in seeds did not enhance longevity. This was deduced from assays of basal thermotolerance and controlled seed-deterioration, which were performed with transgenic tobacco. Furthermore, the constitutive overexpression of HaDREB2 did not increase thermotolerance in seedlings or result in the accumulation of HSPs at normal growth temperatures. In contrast, when HaDREB2 and HaHSFA9 were conjointly overexpressed in seeds, we observed positive effects on seed longevity, beyond those observed with overexpression of HaHSFA9 alone. Such additional effects are accompanied by a subtle enhancement of the accumulation of subsets of sHSPs belonging to the CI and CII cytosolic classes. Our results reveal the functional interdependency of HaDREB2 and HaHSFA9 in seeds. HaDREB2 differs from other previously characterized DREB2 factors in plants in terms of its unique functional interaction with the seed-specific HaHSFA9 factor. No functional interaction between HaDREB2 and HaHSFA9 was observed when both factors were conjointly overexpressed in vegetative tissues. We therefore suggest that additional, seed-specific factors, or protein modifications, could be required for the functional interaction between HaDREB2 and HaHSFA9.

  1. The HaDREB2 transcription factor enhances basal thermotolerance and longevity of seeds through functional interaction with HaHSFA9

    Directory of Open Access Journals (Sweden)

    Carranco Raúl

    2009-06-01

    Full Text Available Abstract Background Transcription factor HaDREB2 was identified in sunflower (Helianthus annuus L. as a drought-responsive element-binding factor 2 (DREB2 with unique properties. HaDREB2 and the sunflower Heat Shock Factor A9 (HaHSFA9 co-activated the Hahsp17.6G1 promoter in sunflower embryos. Both factors could be involved in transcriptional co-activation of additional small heat stress protein (sHSP promoters, and thus contribute to the HaHSFA9-mediated enhancement of longevity and basal thermotolerance of seeds. Results We found that overexpression of HaDREB2 in seeds did not enhance longevity. This was deduced from assays of basal thermotolerance and controlled seed-deterioration, which were performed with transgenic tobacco. Furthermore, the constitutive overexpression of HaDREB2 did not increase thermotolerance in seedlings or result in the accumulation of HSPs at normal growth temperatures. In contrast, when HaDREB2 and HaHSFA9 were conjointly overexpressed in seeds, we observed positive effects on seed longevity, beyond those observed with overexpression of HaHSFA9 alone. Such additional effects are accompanied by a subtle enhancement of the accumulation of subsets of sHSPs belonging to the CI and CII cytosolic classes. Conclusion Our results reveal the functional interdependency of HaDREB2 and HaHSFA9 in seeds. HaDREB2 differs from other previously characterized DREB2 factors in plants in terms of its unique functional interaction with the seed-specific HaHSFA9 factor. No functional interaction between HaDREB2 and HaHSFA9 was observed when both factors were conjointly overexpressed in vegetative tissues. We therefore suggest that additional, seed-specific factors, or protein modifications, could be required for the functional interaction between HaDREB2 and HaHSFA9.

  2. Microscale electrodes integrated on COP for real sample Campylobacter spp. detection.

    Science.gov (United States)

    Morant-Miñana, M Carmen; Elizalde, J

    2015-08-15

    Campylobacter spp. are responsible for acute bacterial diseases in human worldwide. Nowadays campilobacteriosis is considered the most common foodborne illness in the European Union. In this paper the first electrochemical genosensor based on thin-film gold electrodes deposited onto Cyclo Olefin Polymer (COP) substrates was fabricated for the detection of Campylobacter spp in food matrices. The sensing element is characterized by several surface techniques and the sensitivity of the biosensor have been studied. A good linear relationship was obtained for the concentrations of PCR amplicon of Campylobacter spp. between 1 and 25 nM with a limit of detection (LOD) of 90 pM. Real samples have been validated with poultry meat samples and results were comparable with the PCR product samples. This is the last step for the fabrication of a Lab on a Chip (LOC), a biodevice integrating DNA sensor technology into microfluidic system, believed to perform an automated and complete assay, including sample preparation, PCR amplification, and electrochemical detection of Campylobacter spp. in raw poultry meat samples. Copyright © 2015 Elsevier B.V. All rights reserved.

  3. Changing risk of environmental Campylobacter exposure with emerging poultry production systems in Ethiopia.

    Science.gov (United States)

    Brena, M C; Mekonnen, Y; Bettridge, J M; Williams, N J; Wigley, P; Sisay Tessema, T; Christley, R M

    2016-02-01

    Campylobacter is a leading cause of diarrhoea, and its presence in chickens is a significant risk for zoonotic infection. Poultry production is becoming increasingly intensive in Ethiopia and is incorporating more high-producing breeds into traditionally managed smallholdings, especially in peri-urban areas. This cross-sectional study sampled 219 household environments in one peri-urban and two rural areas of Ethiopia, and an additional 20 semi-intensive farms in the peri-urban district. Campylobacter was detected by polymerase chain reaction (PCR)-specific assays in 44 samples; 16 of which could be identified as C. jejuni. Flocks in the peri-urban area were at significantly greater odds of detection, including those which only kept indigenous birds under a scavenging system. It was also noted that scavenging flocks of exotic high-production birds (Rhode Island Red) were at slightly greater risk, perhaps as exotic birds are under more stress when kept under traditional management systems. We suggest that changes to the system of chicken production may alter the ecology and epidemiology of Campylobacter in the environment, chickens and people, which may drive emergence of new epidemiological patterns of disease. Further research is needed to determine the extent to which the current management intensification and the distribution programmes of exotic and/or improved indigenous birds may alter Campylobacter epidemiology, ecology and public health risk, before their widespread adoption.

  4. Multiplex polymerase chain reaction for detection of Campylobacter from stool specimen.

    Science.gov (United States)

    Sarkar, S R; Ray, N C; Hossain, M A; Paul, S K; Sarkar, S; Kobayashi, N

    2014-07-01

    This was a study to prospectively evaluate the sensitivity and specificity of multiplex polymerase chain reaction (PCR) to identify Campylobacter. Multiplex polymerase chain reaction (PCR) based on cadF, hipO & asp gene for Campylobacter genus, C. jejuni & C. coli were tested for detection of Campylobacter jejuni & C. coli in naturally infected faecal samples of human. All the samples were subjected to the cultural isolation of organism and biochemical characterization. The samples resulted in the amplification of a DNA fragment of size 400 bp, 500 bp &735 bp in PCR assay. Two hundred faecal samples comprising diarrheal stools, 23(11.5%) could be detected by isolation whereas 24(12.0%) were found positive by PCR. All culture positive cases were positive by PCR and among 01 culture negative case, were positive by PCR. PCR was found to be more sensitive for Campylobacter detection in faecal samples 12.0% as relative to culture isolation which could detect the organism in 11.5% samples. Sensitivity and specificity of PCR were 100% and 99.4% respectively taking Culture as gold standard. The results depicted the superior efficacy of PCR for rapid screening of samples owing to its high sensitivity, specificity and automation potential.

  5. “Limits of Control” – Crucial Parameters for a Reliable Quantification of Viable Campylobacter by Real-Time PCR

    Science.gov (United States)

    Krüger, Nora-Johanna; Buhler, Christiane; Iwobi, Azuka N.; Huber, Ingrid; Ellerbroek, Lüppo; Appel, Bernd; Stingl, Kerstin

    2014-01-01

    The unsuitability of the “CFU” parameter and the usefulness of cultivation-independent quantification of Campylobacter on chicken products, reflecting the actual risk for infection, is increasingly becoming obvious. Recently, real-time PCR methods in combination with the use of DNA intercalators, which block DNA amplification from dead bacteria, have seen wide application. However, much confusion exists in the correct interpretation of such assays. Campylobacter is confronted by oxidative and cold stress outside the intestine. Hence, damage caused by oxidative stress probably represents the most frequent natural death of Campylobacter on food products. Treatment of Campylobacter with peroxide led to complete loss of CFU and to significant entry of any tested DNA intercalator, indicating disruption of membrane integrity. When we transiently altered the metabolic state of Campylobacter by abolishing the proton-motive force or by inhibiting active efflux, CFU was constant but enhanced entry of ethidium bromide (EtBr) was observed. Consistently, ethidium monoazide (EMA) also entered viable Campylobacter, in particular when nutrients for bacterial energization were lacking (in PBS) or when the cells were less metabolically active (in stationary phase). In contrast, propidium iodide (PI) and propidium monoazide (PMA) were excluded from viable bacterial cells, irrespective of their metabolic state. As expected for a diffusion-limited process, the extent of signal reduction from dead cells depended on the temperature, incubation time and concentration of the dyes during staining, prior to crosslinking. Consistently, free protein and/or DNA present in varying amounts in the heterogeneous matrix lowered the concentration of the DNA dyes at the bacterial membrane and led to considerable variation of the residual signal from dead cells. In conclusion, we propose an improved approach, taking into account principles of method variability and recommend the implementation of

  6. The clinical importance of emerging Campylobacter species.

    Science.gov (United States)

    Man, Si Ming

    2011-10-25

    A growing number of Campylobacter species other than C. jejuni and C. coli have been recognized as emerging human and animal pathogens. Although C. jejuni continues to be the leading cause of bacterial gastroenteritis in humans worldwide, advances in molecular biology and development of innovative culture methodologies have led to the detection and isolation of a range of under-recognized and nutritionally fastidious Campylobacter spp., including C. concisus, C. upsaliensis and C. ureolyticus. These emerging Campylobacter spp. have been associated with a range of gastrointestinal diseases, particularly gastroenteritis, IBD and periodontitis. In some instances, infection of the gastrointestinal tract by these bacteria can progress to life-threatening extragastrointestinal diseases. Studies have shown that several emerging Campylobacter spp. have the ability to attach to and invade human intestinal epithelial cells and macrophages, damage intestinal barrier integrity, secrete toxins and strategically evade host immune responses. Members of the Campylobacter genus naturally colonize a wide range of hosts (including pets, farm animals and wild animals) and are frequently found in contaminated food products, which indicates that these bacteria are at risk of zoonotic transmission to humans. This Review presents the latest information on the role and clinical importance of emerging Campylobacter spp. in gastrointestinal health and disease.

  7. 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

  8. The Metabolic Basis of Pollen Thermo-Tolerance: Perspectives for Breeding

    Directory of Open Access Journals (Sweden)

    Marine J. Paupière

    2014-09-01

    Full Text Available Crop production is highly sensitive to elevated temperatures. A rise of a few degrees above the optimum growing temperature can lead to a dramatic yield loss. A predicted increase of 1–3 degrees in the twenty first century urges breeders to develop thermo-tolerant crops which are tolerant to high temperatures. Breeding for thermo-tolerance is a challenge due to the low heritability of this trait. A better understanding of heat stress tolerance and the development of reliable methods to phenotype thermo-tolerance are key factors for a successful breeding approach. Plant reproduction is the most temperature-sensitive process in the plant life cycle. More precisely, pollen quality is strongly affected by heat stress conditions. High temperature leads to a decrease of pollen viability which is directly correlated with a loss of fruit production. The reduction in pollen viability is associated with changes in the level and composition of several (groups of metabolites, which play an important role in pollen development, for example by contributing to pollen nutrition or by providing protection to environmental stresses. This review aims to underline the importance of maintaining metabolite homeostasis during pollen development, in order to produce mature and fertile pollen under high temperature. The review will give an overview of the current state of the art on the role of various pollen metabolites in pollen homeostasis and thermo-tolerance. Their possible use as metabolic markers to assist breeding programs for plant thermo-tolerance will be discussed.

  9. Oxygen availability strongly affects chronological lifespan and thermotolerance in batch cultures of Saccharomyces cerevisiae

    Science.gov (United States)

    Bisschops, Markus M.; Vos, Tim; Martínez-Moreno, Rubén; Cortés, Pilar T.; Pronk, Jack T.; Daran-Lapujade, Pascale

    2015-01-01

    Stationary-phase (SP) batch cultures of Saccharomyces cerevisiae, in which growth has been arrested by carbon-source depletion, are widely applied to study chronological lifespan, quiescence and SP-associated robustness. Based on this type of experiments, typically performed under aerobic conditions, several roles of oxygen in aging have been proposed. However, SP in anaerobic yeast cultures has not been investigated in detail. Here, we use the unique capability of S. cerevisiae to grow in the complete absence of oxygen to directly compare SP in aerobic and anaerobic bioreactor cultures. This comparison revealed strong positive effects of oxygen availability on adenylate energy charge, longevity and thermotolerance during SP. A low thermotolerance of anaerobic batch cultures was already evident during the exponential growth phase and, in contrast to the situation in aerobic cultures, was not substantially increased during transition into SP. A combination of physiological and transcriptome analysis showed that the slow post-diauxic growth phase on ethanol, which precedes SP in aerobic, but not in anaerobic cultures, endowed cells with the time and resources needed for inducing longevity and thermotolerance. When combined with literature data on acquisition of longevity and thermotolerance in retentostat cultures, the present study indicates that the fast transition from glucose excess to SP in anaerobic cultures precludes acquisition of longevity and thermotolerance. Moreover, this study demonstrates the importance of a preceding, calorie-restricted conditioning phase in the acquisition of longevity and stress tolerance in SP yeast cultures, irrespective of oxygen availability. PMID:28357268

  10. Oxygen availability strongly affects chronological lifespan and thermotolerance in batch cultures of Saccharomyces cerevisiae

    Directory of Open Access Journals (Sweden)

    Markus M.M. Bisschops

    2015-10-01

    Full Text Available Stationary-phase (SP batch cultures of Saccharomyces cerevisiae, in which growth has been arrested by carbon-source depletion, are widely applied to study chronological lifespan, quiescence and SP-associated robustness. Based on this type of experiments, typically performed under aerobic conditions, several roles of oxygen in aging have been proposed. However, SP in anaerobic yeast cultures has not been investigated in detail. Here, we use the unique capability of S. cerevisiae to grow in the complete absence of oxygen to directly compare SP in aerobic and anaerobic bioreactor cultures. This comparison revealed strong positive effects of oxygen availability on adenylate energy charge, longevity and thermotolerance during SP. A low thermotolerance of anaerobic batch cultures was already evident during the exponential growth phase and, in contrast to the situation in aerobic cultures, was not substantially increased during transition into SP. A combination of physiological and transcriptome analysis showed that the slow post-diauxic growth phase on ethanol, which precedes SP in aerobic, but not in anaerobic cultures, endowed cells with the time and resources needed for inducing longevity and thermotolerance. When combined with literature data on acquisition of longevity and thermotolerance in retentostat cultures, the present study indicates that the fast transition from glucose excess to SP in anaerobic cultures precludes acquisition of longevity and thermotolerance. Moreover, this study demonstrates the importance of a preceding, calorie-restricted conditioning phase in the acquisition of longevity and stress tolerance in SP yeast cultures, irrespective of oxygen availability.

  11. Thermotolerance in preirradiated intestine and its influence on time-temperature relationships

    International Nuclear Information System (INIS)

    Hume, S.P.; Marigold, J.C.; Manjil, L.G.

    1988-01-01

    The crypt compartment of mouse jejunum showed a transient increase in thermal susceptibility approximately 10 days after moderate X-ray doses to the abdomen (9-10 Gy). The increase in response was manifest as an increase in slope of the crypt dose-response curve but was limited to temperatures below 43 0 C. As a result, the 43 0 C inflexion in the Arrhenius plot (the relationship between treatment time and temperature) for thermal sensitivity of crypts was eliminated in preirradiated tissue, and the curve became monophasic over the range 42.0-44.5 0 C. At temperatures below 42 0 C, the curve again deviated. At supranormal temperatures of 42 0 C and below, the durations of hyperthermia needed for measurable effect were sufficient to allow thermotolerance to be expressed within the heating period. Neither the threshold heating times nor this thermotolerance were affected by prior irradiation. In the temperature range 42-43 0 C, an earlier development of thermotolerance could be demonstrated in control tissue by challenging with an acute high-temperature heat treatment. This thermotolerance was eliminated in preirradiated tissue, resulting in the apparent increase in sensitivity. The findings support the view that the complex nature of the time-temperature relationship seen in normal tissue in vivo is a manifestation of the ability of the tissue to progressively acquire a thermotolerant state during treatment at temperatures below approximately 43 0 C, so that the intrinsic sensitivity is modulated while being assessed

  12. Robustness promotes evolvability of thermotolerance in an RNA virus

    Directory of Open Access Journals (Sweden)

    Turner Paul E

    2008-08-01

    Full Text Available Abstract Background The ability for an evolving population to adapt to a novel environment is achieved through a balance of robustness and evolvability. Robustness is the invariance of phenotype in the face of perturbation and evolvability is the capacity to adapt in response to selection. Genetic robustness has been posited, depending on the underlying mechanism, to either decrease the efficacy of selection, or increase the possibility of future adaptation. However, the true effect of genetic robustness on evolvability in biological systems remains uncertain. Results Here we demonstrate that genetic robustness increases evolvability of thermotolerance in laboratory populations of the RNA virus φ6. We observed that populations founded by robust clones evolved greater resistance to heat shock, relative to populations founded by brittle (less-robust clones. Thus, we provide empirical evidence for the idea that robustness can promote evolvability in this environment, and further suggest that evolvability can arise indirectly via selection for robustness, rather than through direct selective action. Conclusion Our data imply that greater tolerance of mutational change is associated with virus adaptability in a new niche, a finding generally relevant to evolutionary biology, and informative for elucidating how viruses might evolve to emerge in new habitats and/or overcome novel therapies.

  13. Directed evolution of thermotolerant malic enzyme for improved malate production.

    Science.gov (United States)

    Morimoto, Yumi; Honda, Kohsuke; Ye, Xiaoting; Okano, Kenji; Ohtake, Hisao

    2014-02-01

    The directed evolution of the thermotolerant NADP(H)-dependent malic enzyme from Thermococcus kodakarensis was conducted to alter the cofactor preference of the enzyme from NADP(H) to NAD(H). The construction and screening of two generations of mutant libraries led to the isolation of a triple mutant that exhibited 6-fold higher kcat/Km with NAD(+) than the wild type. We serendipitously found that, in addition to the change in the cofactor preference, the reaction specificity of the mutant enzyme was altered. The reductive carboxylation of pyruvate to malate catalyzed by the wild type enzyme is accompanied by HCO(3)(-)-independent reduction of pyruvate and gives lactate as a byproduct. The reaction specificity of the triple mutant was significantly shifted to malate production and the mutant gave a less amount of the byproduct than the wild type. When the triple mutant enzyme was used as a catalyst for pyruvate carboxylation with NADH, the enzyme gave 1.2 times higher concentration of malate than the wild type with NADPH. Single-point mutation analysis revealed that the substitution of Arg221 with Gly is responsible for the shift in reaction specificity. This finding may shed light on the catalytic mechanisms of malic enzymes and other related CO2- and/or HCO(3)(-)-fixing enzymes. Copyright © 2013 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  14. Selective medium for growth of Campylobacter in containers incubated aerobically

    Science.gov (United States)

    Introduction. Campylobacter are traditionally cultured in primary containers inside of secondary containers filled with microaerobic atmospheres. Recent findings indicated that media supplemented with optimal concentrations of amino acids, organic acids, and bicarbonate support Campylobacter growth ...

  15. Campylobacter bacteremia: a rare and under-reported event?

    Science.gov (United States)

    Louwen, R.; van Baarlen, P.; van Vliet, A. H. M.; van Belkum, A.; Hays, J. P.; Endtz, H. P.

    2012-01-01

    Bacteria belonging to the species Campylobacter are the most common cause of bacterial diarrhoea in humans. The clinical phenotype associated with Campylobacter infections ranges from asymptomatic conditions to severe colitis and bacteremia. In susceptible patients, Campylobacter infections are associated with significant morbidity and mortality, with both host factors and bacterial factors being involved in the pathogenesis of bacteremia. In the host, age, gender and immune-compromising conditions may predispose for Campylobacter infections, whilst the most important bacterial determinants mentioned in the literature are cytotoxin production and flagellar motility. The role of sialylated lipo-oligosaccharide (LOS) and serum resistance in bacteremia is inconclusive at this time, and the clinical significance of Campylobacter bacteremia is not yet fully understood. More emphasis on the detection of Campylobacter species from blood cultures in susceptible patients at risk for Campylobacter infections will increase our understanding of the pathogenesis and the relevance of Campylobacter bacteremia. PMID:24611124

  16. 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...

  17. Salicylate Functions as an Efflux Pump Inducer and Promotes the Emergence of Fluoroquinolone-Resistant Campylobacter jejuni Mutants▿

    Science.gov (United States)

    Shen, Zhangqi; Pu, Xiao-Ying; Zhang, Qijing

    2011-01-01

    Salicylate, a nonsteroidal anti-inflammatory compound, has been shown to increase the resistance of Campylobacter to antimicrobials. However, the molecular mechanism underlying salicylate-induced resistance has not yet been established. In this study, we determined how salicylate increases antibiotic resistance and evaluated its impact on the development of fluoroquinolone-resistant Campylobacter mutants. Transcriptional fusion assays, real-time quantitative reverse transcription-PCR (RT-PCR), and immunoblotting assays consistently demonstrated the induction of the CmeABC multidrug efflux pump by salicylate. Electrophoretic mobility shift assays further showed that salicylate inhibits the binding of CmeR (a transcriptional repressor of the TetR family) to the promoter DNA of cmeABC, suggesting that salicylate inhibits the function of CmeR. The presence of salicylate in the culture medium not only decreased the susceptibility of Campylobacter to ciprofloxacin but also resulted in an approximately 70-fold increase in the observed frequency of emergence of fluoroquinolone-resistant mutants under selection with ciprofloxacin. Together, these results indicate that in Campylobacter, salicylate inhibits the binding of CmeR to the promoter DNA and induces expression of cmeABC, resulting in decreased susceptibility to antibiotics and in increased emergence of fluoroquinolone-resistant mutants under selection pressure. PMID:21821741

  18. Detection of Campylobacter spp. in chilled and frozen broiler carcasses comparing immunoassay, PCR and real time PCR methods

    Directory of Open Access Journals (Sweden)

    Luciana Pimenta Reis

    2018-02-01

    Full Text Available ABSTRACT: In order to detect and identify Campylobacter spp. in broiler chicken carcasses, and to compare detection methods, 43 chilled and 43 frozen carcasses were collected and analyzed. Three methodologies were evaluated: an automated Enzyme Linked Fluorescent Assay (ELFA VIDAS®30, Polymerase Chain Reaction (PCR and real-time PCR. Only four chilled carcasses (4.6% were considered positive for Campylobacter spp. by VIDAS®30 and no sample was positive when the conventional PCR technique was used. However, real-time PCR showed a higher incidence of contamination by Campylobacter spp. in broiler carcasses, with 45 (52.3% positive samples. C. jejuni was the species most frequently reported in the samples (88.8%. No differences in the frequencies of Campylobacter spp. were observed between the chilled and frozen broiler carcasses. In conclusion, real-time PCR was the most sensitive method for the detection of Campylobacter spp. in chilled or frozen broiler carcasses, which were mainly contaminated by C. jejuni.

  19. 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).

  20. 21 CFR 866.3110 - Campylobacter fetus serological reagents.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Campylobacter fetus serological reagents. 866.3110... (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3110 Campylobacter fetus serological reagents. (a) Identification. Campylobacter fetus serological reagents are devices...

  1. 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

  2. Enhanced lipid production in thermo-tolerant mutants of Chlorella pyrenoidosa NCIM 2738.

    Science.gov (United States)

    Sachdeva, Neha; Gupta, Ravi Prakash; Mathur, Anshu Shankar; Tuli, Deepak Kumar

    2016-12-01

    The present study aimed to develop thermo-tolerant mutants of Chlorella pyrenoidosa NCIM 2738 for high lipids production. For this, ethyl methane sulfonate was used, which generated two effective thermo-tolerant mutants, M18 and M24 of Chlorella pyrenoidosa NCIM 2738, capable of surviving at temperature up to 47°C and showing improved lipid and biomass yields. They showed 59.62% and 50.75% increase, respectively in lipid content compared to wild type at 30°C, which could not grow at temperature above 35°C. The novelty of this study lied in incorporation of PAM Flurometry with mutagenesis to generate thermo-tolerant mutants of C. pyrenoidosa and investigating the reasons for increased yields of mutants at cellular and photosynthetic levels with the aim to use them for commercial biodiesel production. Copyright © 2016 Elsevier Ltd. All rights reserved.

  3. 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.

  4. Heat-treated Campylobacter spp. and mRNA stability as determined by reverse transcriptase-polymerase chain reaction.

    Science.gov (United States)

    Sung, Kidon; Hiett, Kelli L; Stern, Norman J

    2005-01-01

    The detection method of reverse transcriptase-polymerase chain reaction (RT-PCR), which specifically targets mRNA, was developed and tested for detection of cultivable Campylobacter spp. The expression of four DNA targets- flaA, tkt, porA, and a putative haem-copper oxidase domain-were assayed in heat-inactivated Campylobacter spp. to determine an optimum target for RT-PCR amplification. A diversity of Campylobacter spp. was tested; however, the presented RT-PCR technique was specific for C. jejuni, C. coli, and C. lari. The durability of mRNA species detected by our RT-PCR technique was dependent upon the individual Campylobacter spp. examined, the condition of heat treatment and post-treatment holding time, as well as the transcript targeted. The putative oxidase was determined to be the most stable mRNA species for this assay. The mRNA of the putative oxidase gene was detectable even after Campylobacter spp. had been treated at temperatures of 95-99 degrees C. Using DNA-based PCR, the four DNA targets could be amplified after heat inactivation followed by a 48 h holding time, indicating that the chromosomal DNA was not substantially degraded by the heat treatment. PCR products from the putative oxidase gene were detected at 10(2) to 10(3) C. jejuni CFU per mL, exhibiting the highest level of sensitivity among the genes tested. The results in the present study indicate that mRNA from Campylobacter spp. may persist in a form that is detectable by RT-PCR amplification for an extended period after heat treatment, demonstrating a poor correlation between mRNA detection and cell cultivability. With these results in mind, further investigations are necessary to determine the correlation between RT-PCR amplification and viability.

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

    Directory of Open Access Journals (Sweden)

    Tambur Zoran Ž.

    2010-01-01

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

  6. 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...

  7. Enhancing the Thermotolerance of Entomopathogenic Isaria fumosorosea SFP-198 Conidial Powder by Controlling the Moisture Content Using Drying and Adjuvants.

    Science.gov (United States)

    Kim, Jae Su; Lee, Se Jin; Lee, Hyang Burm

    2014-03-01

    Entomopathogenic fungi are promising pest-control agents but their industrial applicability is limited by their thermosusceptibility. With an aim to increase the thermotolerance of Isaria fumosorosea SFP-198, moisture absorbents were added to dried conidial powder, and the relationship between its water potential and thermotolerance was investigated. Mycotized rice grains were dried at 10℃, 20℃, 30℃, and 40℃ and the drying effect of each temperature for 24, 48, 96, and 140 hr was determined. Drying for 48 hr at 10℃ and 20℃ reduced the moisture content to < 5% without any significant loss of conidial thermotolerance, but drying at 30℃ and 40℃ reduced both moisture content and conidial thermotolerance. To maintain thermotolerance during storage, moisture absorbents, such as calcium chloride, silica gel, magnesium sulfate, white carbon, and sodium sulfate were individually added to previously dried-conidial powder at 10% (w/w). These mixtures was then stored at room temperature for 30 days and subjected to 50℃ for 2 hr. The white carbon mixture had the highest conidial thermotolerance, followed by silica gel, magnesium sulfate, and then the other absorbents. A significant correlation between the water potential and conidial thermotolerance was observed in all conidia-absorbent mixtures tested in this study (r = -0.945). Conidial thermotolerance in wet conditions was evaluated by adding moisturized white carbon (0~20% H2O) to conidia to mimic wet conditions. Notably, the conidia still maintained their thermotolerance under these conditions. Thus, it is evident that conidial thermotolerance can be maintained by drying mycotized rice grains at low temperatures and adding a moisture absorbent, such as white carbon.

  8. Characterization of Campylobacter phages including analysis of host range by selected Campylobacter Penner serotypes

    Directory of Open Access Journals (Sweden)

    Brown Stanley

    2007-10-01

    Full Text Available Abstract Background The predominant food borne pathogen in the western world today is Campylobacter. Campylobacter specific bacteriophages (phages have been proposed as an alternative agent for reducing the burden of Campylobacter in broilers. One concern in relation to phage biocontrol is the narrow host range often displayed by phages. To identify the potential of phages as a Campylobacter reducing agent we needed to determine their infectivity on a panel of isolates representing the Campylobacter strains found in broilers as well as humans. Results In this study, Campylobacter phages were isolated from the intestines of broilers and ducks and from abattoir sewage. Twelve phages were investigated to determine their ability to infect the Campylobacter Penner serotypes commonly present in Danish poultry and patients with campylobacteriosis. A total of 89% of the Campylobacter jejuni strains and 14% of the Campylobacter coli strains could be infected by at least one of the bacteriophages. The majority of the phages infected the most common serotypes in Danish broilers (O:1,44; O:2; O:4-complex, but showed limited ability to infect 21 of the less frequent Campylobacter serotypes. Pulse field gel electrophoresis (PFGE and restriction endonuclease analysis (REA were used to characterize the phage genomes. Three categories of bacteriophages were observed. I: a genome size of ~194 kb and refractory to digestion with HhaI; II: a genome size of ~140 kb and digestible by HhaI; and III: a genome size undeterminable in PFGE. The categorization of the phages correlated with the host range patterns displayed by the phages. Six phages were subjected to transmission electron microscopy (TEM. They all belonged to the family of Myoviridae. Conclusion We have characterized and identified the host range of 12 Danish Campylobacter phages. Due to their ability to infect the majority of the common serotypes in Denmark we suggest the phages can become an effective

  9. Characterization of Campylobacter phages including analysis of host range by selected Campylobacter Penner serotypes.

    Science.gov (United States)

    Hansen, Vinni Mona; Rosenquist, Hanne; Baggesen, Dorte Lau; Brown, Stanley; Christensen, Bjarke Bak

    2007-10-18

    The predominant food borne pathogen in the western world today is Campylobacter. Campylobacter specific bacteriophages (phages) have been proposed as an alternative agent for reducing the burden of Campylobacter in broilers. One concern in relation to phage biocontrol is the narrow host range often displayed by phages. To identify the potential of phages as a Campylobacter reducing agent we needed to determine their infectivity on a panel of isolates representing the Campylobacter strains found in broilers as well as humans. In this study, Campylobacter phages were isolated from the intestines of broilers and ducks and from abattoir sewage. Twelve phages were investigated to determine their ability to infect the Campylobacter Penner serotypes commonly present in Danish poultry and patients with campylobacteriosis. A total of 89% of the Campylobacter jejuni strains and 14% of the Campylobacter coli strains could be infected by at least one of the bacteriophages. The majority of the phages infected the most common serotypes in Danish broilers (O:1,44; O:2; O:4-complex), but showed limited ability to infect 21 of the less frequent Campylobacter serotypes. Pulse field gel electrophoresis (PFGE) and restriction endonuclease analysis (REA) were used to characterize the phage genomes. Three categories of bacteriophages were observed. I: a genome size of approximately 194 kb and refractory to digestion with HhaI; II: a genome size of approximately 140 kb and digestible by HhaI; and III: a genome size undeterminable in PFGE. The categorization of the phages correlated with the host range patterns displayed by the phages. Six phages were subjected to transmission electron microscopy (TEM). They all belonged to the family of Myoviridae. We have characterized and identified the host range of 12 Danish Campylobacter phages. Due to their ability to infect the majority of the common serotypes in Denmark we suggest the phages can become an effective agent in the effort to reduce

  10. Microarray on digital versatile disc for identification and genotyping of Salmonella and Campylobacter in meat products.

    Science.gov (United States)

    Tortajada-Genaro, Luis Antonio; Rodrigo, Alejandro; Hevia, Elizabeth; Mena, Salvador; Niñoles, Regina; Maquieira, Ángel

    2015-09-01

    Highly portable, cost-effective, and rapid-response devices are required for the subtyping of the most frequent food-borne bacteria; thereby the sample rejection strategies and hygienization techniques along the food chain can be tailor-designed. Here, a novel biosensor is presented for the generic detection of Salmonella and Campylobacter and the discrimination between their most prevalent serovars (Salmonella Enteritidis, Salmonella Typhimurium) and species (Campylobacter jejuni, Campylobacter coli), respectively. The method is based on DNA microarray developed on a standard digital versatile disc (DVD) as support for a hybridization assay and a DVD driver as scanner. This approach was found to be highly sensitive (detection limit down to 0.2 pg of genomic DNA), reproducible (relative standard deviation 4-19 %), and high working capacity (20 samples per disc). The inclusivity and exclusivity assays indicated that designed oligonucleotides (primers and probes) were able to discriminate targeted pathogens from other Salmonella serovars, Campylobacter species, or common food-borne pathogens potentially present in the indigenous microflora. One hundred isolates from meat samples, collected in a poultry factory, were analyzed by the DVD microarraying and fluorescent real-time PCR. An excellent correlation was observed for both generic and specific detection (relative sensitivity 93-99 % and relative specificity 93-100 %). Therefore, the developed assay has been shown to be a reliable tool to be used in routine food safety analysis, especially in settings with limited infrastructure due to the excellent efficiency-cost ratio of compact disc technology. Graphical Abstract DNA microarray performed by DVD technology for pathogen genotyping.

  11. The Prevalence of Multiple Antibiotic Resistance in Campylobacter spp. From Retail Poultry Meat

    Directory of Open Access Journals (Sweden)

    Marija Kurinčič

    2005-01-01

    Full Text Available Macrolides and fluoroquinolones are regarded as drugs of choice for the treatment of human Campylobacter infections. The use of antimicrobials for this purpose as well as in food animal production has resulted in the resistance of Campylobacter spp. to selected antibiotics. Since poultry is one of the most important sources of human Campylobacter infections the use of antibiotics in animal production can shorten the effective therapeutic life of antibiotics for human use. During 2001–2003, over 220 strains of C. jejuni and C. coli were isolated from 60 poultry meat samples from the retail market in Slovenia and further characterized by phenotypic and molecular methods. In this study, 55 sample-representative strains were tested for susceptibility to eight different antibiotics (ampicillin, amoxycillin/ clavulanic acid, ciprofloxacin, erythromycin, gentamicin, nalidixic acid, pefloxacin and tetracycline. Phenotypic procedures (disc diffusion test, E-test as well as molecular detection of mutations (mismatch amplification mutation assay (MAMA polymerase chain reaction (PCR in case of ciprofloxacin resistance were used. When assuming the results about antibiotic resistance, only 38.2 % of strains tested were susceptible to all antibiotics tested. Regarding ciprofloxacin, 58.2 % of tested strains were found to be resistant (minimal inhibitory concentration, MIC>4 mg/mL. The occurrence of resistance was much higher in C. coli (75.9 % than in C. jejuni (38.5 % isolates. The resistance rates to pefloxacin, nalidixic acid, erythromycin and tetracycline were 58.2, 49.1, 14.5 and 12.7 %, respectively. Eleven percent of strains were resistant to erythromycin and ciprofloxacin and 12.7 % of strains were resistant to tetracycline and quinolones. The results show the need for monitoring the prevalence and antibiotic resistance of zoonotic bacteria such as Campylobacter as well as the multiresistance phenomenon of Campylobacter isolates from food in our

  12. Thermotolerance of an inactivated rabies vaccine for dogs.

    Science.gov (United States)

    Lankester, Felix J; Wouters, Pieter A W M; Czupryna, Anna; Palmer, Guy H; Mzimbiri, Imam; Cleaveland, Sarah; Francis, Mike J; Sutton, David J; Sonnemans, Denny G P

    2016-11-04

    This study provides the first robust data that the antibody response of dogs vaccinated with Nobivac® Rabies vaccine stored for several months at high temperatures (up to 30°C) is not inferior to that of dogs vaccinated with vaccine stored under recommended cold-chain conditions (2-8°C). A controlled and randomized non-inferiority study was carried out comparing the four-week post vaccination serological responses of Tanzanian village dogs inoculated with vaccine which had been stored at elevated temperatures for different periods of time with those of dogs vaccinated with the same product stored according to label recommendations. Specifically, the neutralizing antibody response following the use of vaccine which had been stored for up to six months at 25°C or for three months at 30°C was not inferior to that following the use of cold-chain stored vaccine. These findings provide reassurance that the vaccine is likely to remain efficacious even if exposed to elevated temperatures for limited periods of time and, under these circumstances, it can safely be used and not necessarily destroyed or discarded. The availability of thermotolerant vaccines has been an important factor in the success of several disease control and elimination programs and could greatly increase the capacity of rabies vaccination campaigns to access hard to reach communities in Africa and Asia. We have not confirmed a 3-year duration of immunity for the high temperature stored vaccine, however because annual re-vaccination is usually practiced for dogs presented for vaccination during campaigns in Africa and Asia this should not be a cause for concern. These findings will provide confidence that, for rabies control and elimination programs using this vaccine in low-income settings, more flexible delivery models could be explored, including those that involve limited periods of transportation and storage at temperatures higher than that currently recommended. Copyright © 2016 The Authors

  13. Characterization of Campylobacter phages including analysis of host range by selected Campylobacter Penner serotypes

    DEFF Research Database (Denmark)

    Hansen, Vinni; Rosenquist, Hanne; Baggesen, Dorte Lau

    2007-01-01

    size undeterminable in PFGE. The categorization of the phages correlated with the host range patterns displayed by the phages. Six phages were subjected to transmission electron microscopy (TEM). They all belonged to the family of Myoviridae. Conclusion: We have characterized and identified the host......Background: The predominant food borne pathogen in the western world today is Campylobacter. Campylobacter specific bacteriophages (phages) have been proposed as an alternative agent for reducing the burden of Campylobacter in broilers. One concern in relation to phage biocontrol is the narrow host...

  14. 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...

  15. Faecal Campylobacter shedding among dogs in animal shelters across Texas.

    Science.gov (United States)

    Leahy, A M; Cummings, K J; Rodriguez-Rivera, L D; Hamer, S A; Lawhon, S D

    2017-12-01

    Epidemiologic studies on faecal Campylobacter shedding among dogs in the United States have been limited, despite evidence that the incidence of human campylobacteriosis has increased over the last decade. Our objectives were to estimate the prevalence of faecal Campylobacter shedding among shelter dogs in Texas, to estimate the specific prevalence of Campylobacter jejuni and Campylobacter coli shedding, and to identify risk factors for Campylobacter-positive status. Using a cross-sectional study design, we collected faecal samples from dogs in six animal shelters across Texas between May and December, 2014. Quantitative PCR protocols were used to detect Campylobacter in samples and to specifically identify C. jejuni and C. coli. The prevalence of faecal Campylobacter shedding among sampled dogs was 75.7% (140/185). Prevalence varied significantly by shelter (p = .03), ranging from 57% to 93%. There was a marginal association (p = .06) between abnormal faecal consistency and positive Campylobacter status, after controlling for shelter as a random effect. However, approximately 70% of Campylobacter-positive dogs had grossly normal faeces. Campylobacter prevalence did not vary significantly by age group or sex. The prevalence of C. jejuni-positive samples was 5.4% (10/185), but C. coli was not detected in any samples. Dogs are a potential source of zoonotic Campylobacter transmission. © 2017 Blackwell Verlag GmbH.

  16. Multiplex real-time PCR for detection of Campylobacter, Salmonella, and Shigella.

    Science.gov (United States)

    Barletta, F; Mercado, E H; Lluque, A; Ruiz, J; Cleary, T G; Ochoa, T J

    2013-09-01

    Infectious diarrhea can be classified based on its clinical presentation as noninflammatory or inflammatory disease. In developing countries, among inflammatory diarrhea cases, Shigella is the most common cause, followed by Campylobacter and Salmonella. Because the time frame in which treatment choices must be made is short and conventional stool cultures lack good sensitivity, there is a need for a rapid, sensitive, and inexpensive detection technique. The purpose of our study was to develop a multiplex real-time PCR procedure to simultaneously identify Campylobacter spp., Salmonella spp., and Shigella spp. Primers were designed to amplify the invA, ipaH, and 16S rRNA genes simultaneously in a single reaction to detect Salmonella, Shigella, and Campylobacter, respectively. Using this approach, we correctly identified 102 of 103 strains of the targeted enteropathogens and 34 of 34 other pathogens. The melting temperatures were 82.96 ± 0.05 °C for invA, 85.56 ± 0.28 °C for ipaH, and 89.21 ± 0.24 °C for 16S rRNA. The limit of accurate quantification for the assay in stool samples was 10(4) CFU g(-1); however, the limit of detection was 10(3) CFU g(-1). This assay is a simple, rapid, inexpensive, and reliable system for the practical detection of these three enteropathogens in clinical specimens.

  17. Metode Direct Polymerase Chain Reaction untuk Melacak Campylobacter sp. pada Daging Ayam (DIRECT POLYMERASE CHAIN REACTION METHOD FOR DETECTION CAMPYLOBACTER SP. OF POULTRY MEAT

    Directory of Open Access Journals (Sweden)

    Andriani .

    2013-08-01

    Full Text Available Campylobacter sp. is the most commonly reported as agent of foodborne zoonosis causing acutegastroenteritis in humans. Poultry meat is considered as a major source of C. jejuni infection in human.The conventional methods for detecting foodborne bacteria is time-consuming which rely on the of thebacteria in culture media, followed by biochemical identification. In this study polymerase chain reaction(PCR technique was used for rapid identification of the pathogenic Campylobacter sp. The samples usedwere 298 chicken carcass with sold in supermarkets and traditional markets, and were carried out inaccordance the isolation protocol ISO/ DIS 10272-1994. Identification was performed using biochemicalAPI Campy. The direct PCR (DPCR assay with two sets of primers was employed for isolation andidentification of C. jejuni and C. coli. The result of the isolation and identification both by conventional orPCR methods showed that chicken carcasses both from supermarket and traditional market werecontaminated with C. jejuni and or C. coli. Prevalence of Campylobacter sp. contamination in chicken meatwas higher by DPCR (62.6% than by conventional (19.8%, indicating that DPCR technique was moresensitive than conventional method with detection limit for C. jejuni was103 cfu/ml.

  18. 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...

  19. Foodborne Campylobacter: Infections, Metabolism, Pathogenesis and Reservoirs

    Science.gov (United States)

    Epps, Sharon V. R.; Harvey, Roger B.; Hume, Michael E.; Phillips, Timothy D.; Anderson, Robin C.; Nisbet, David J.

    2013-01-01

    Campylobacter species are a leading cause of bacterial-derived foodborne illnesses worldwide. The emergence of this bacterial group as a significant causative agent of human disease and their propensity to carry antibiotic resistance elements that allows them to resist antibacterial therapy make them a serious public health threat. Campylobacter jejuni and Campylobacter coli are considered to be the most important enteropathogens of this genus and their ability to colonize and survive in a wide variety of animal species and habitats make them extremely difficult to control. This article reviews the historical and emerging importance of this bacterial group and addresses aspects of the human infections they cause, their metabolism and pathogenesis, and their natural reservoirs in order to address the need for appropriate food safety regulations and interventions. PMID:24287853

  20. Campylobacter ureolyticus: an emerging gastrointestinal pathogen?

    Science.gov (United States)

    Bullman, Susan; Corcoran, Daniel; O'Leary, James; Lucey, Brigid; Byrne, Deirdre; Sleator, Roy D

    2011-03-01

    A total of 7194 faecal samples collected over a 1-year period from patients presenting with diarrhoea were screened for Campylobacter spp. using EntericBio(®) , a multiplex-PCR system. Of 349 Campylobacter-positive samples, 23.8% were shown to be Campylobacter ureolyticus, using a combination of 16S rRNA gene analysis and highly specific primers targeting the HSP60 gene of this organism. This is, to the best of our knowledge, the first report of C. ureolyticus in the faeces of patients presenting with gastroenteritis and may suggest a role for this organism as an emerging enteric pathogen. © 2010 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.

  1. Pleurotus sajor-caju HSP100 complements a thermotolerance defect in hsp104 mutant Saccharomyces cerevisiae.

    Science.gov (United States)

    Lee, Jin-Ohk; Jeong, Mi-Jeong; Kwon, Tack-Ryun; Lee, Seung-Kon; Byun, Myung-Ok; Chung, Ill-Min; Park, Soo-Chul

    2006-06-01

    A putative Hsp100 gene was cloned from the fungus Pleurotus sajor-caju. mRNA expression studies demonstrated that this gene (designated PsHsp100) is highly induced by high temperature,induced less strongly by exposure to ethanol, and not induced by drought or salinity. Heat shock induction is detectable at 37 degrees C and reaches a maximum level at 42 degrees C. PsHsp100 mRNA levels sharply increased within 15 min of exposure to high temperature, and reached a maximum expression level at 2 h that was maintained for several hours. These results indicate that PsHsp100 could work at an early step in thermotolerance. To examine its function, PsHsp100 was transformed into a temperature-sensitive hsp104 deletion mutant Saccharomycetes cerivisiae strain to test the hypothesis that PsHSP100 is an protein that functions in thermotolerance. Overexpression of PsHSP100 complemented the thermotolerance defect of the hsp104 mutant yeast, allowing them being survive even at 50 degree C for 4 h. These results indicate that PsHSP100 protein is functional as an HSP100 in yeast and could play and important role in thermotolerance in P. sajor-caju.

  2. 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...

  3. 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.

  4. Campylobacter gastroenteritis associated with Sweet's syndrome.

    Science.gov (United States)

    Pai, Sumita; Rytina, Ed; Sterling, Jane; Karas, J A; Aliyu, S H

    2012-10-01

    Sweet's syndrome or acute febrile neutrophilic dermatosis has been associated with underlying infection, malignancy, inflammatory disease and certain medications. The infection agents associated with this include Streptococcus species, Yersinia species, Chlamydia species, Salmonella species and Helicobacter pylori. We report a case of Sweet's syndrome in a 73-year-old woman following a 2 week course of severe gastroenteritis caused by Campylobacter species. Histological examination of skin lesions showed marked inflammatory infiltrate throughout the dermis, composed of neutrophils and histiocytes. The patient was successfully treated with topical and systemic steroids. To date, this is the first case of Sweet's syndrome to be reported linked to Campylobacter species to our knowledge.

  5. Development of transient phage resistance in Campylobacter coli against the group II phage CP84.

    Science.gov (United States)

    Orquera, Stefanie; Hertwig, Stefan; Alter, Thomas; Hammerl, Jens A; Jirova, Alice; Gölz, Greta

    2015-01-01

    Recently, there is a growing interest in the use of bacteriophages for pre- and post-harvest applications to reduce foodborne pathogens (including Campylobacter) along the food chain. Quantitative Campylobacter reductions of up to three log10 units have been achieved by phage application. However, possible phage resistance might limit this approach. In Campylobacter (C.) jejuni, phage resistance mechanisms have been described in detail but data on these mechanisms in C. coli are still missing. To study phage resistance in C. coli, strain NCTC 12668 was infected with the lytic phage CP84, belonging to group II of Campylobacter phages. Resistant and sensitive clones were analysed using phenotypic and genotypic assays. C. coli clones acquired only transient resistance against CP84. The resistance led to cross-protection to one out of five other group II phages tested. Phage resistance was apparently neither caused by large genomic rearrangements nor by a CRISPR system. Binding assays demonstrated that CP84 could not adsorb to resistant C. coli clones suggesting a bacterial phage receptor to be involved in resistance. However, phage resistant C. coli clones did not reveal an altered motility or modified flaA sequence. Considering the loss of binding capacity and the reversion to a phage sensitive phenotype we hypothesize that acquired resistance depends on temporal phase variable switch-off modifications of the phage receptor genes, even though the resistance mechanism could not be elucidated in detail. We further speculate that even closely related phages of the same group use different bacterial receptors for binding on C. coli.

  6. 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...

  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

    , 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...

  8. 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.

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

    DEFF Research Database (Denmark)

    Hald, Birthe; Madsen, Mogens

    1997-01-01

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

  10. 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...

  11. National surveillance of Campylobacter in broilers at slaughter in Denmark in 1998

    DEFF Research Database (Denmark)

    Wedderkopp, A.; Rattenborg, Erik; Madsen, Mogens

    2000-01-01

    was Campylobacter jejuni 86%, Campylobacter coli 11%, Campylobacter lari 1%, other not further diagnosed species 2%. The prevalence was significantly higher in the period from June to October (3.2 abattoir (OR

  12. 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.

  13. 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.

  14. Campylobacter Fetus Meningitis in Adults

    Science.gov (United States)

    van Samkar, Anusha; Brouwer, Matthijs C.; van der Ende, Arie; van de Beek, Diederik

    2016-01-01

    Abstract The zoonotic pathogen Campylobacter fetus is a rare cause of bacterial meningitis. Little is known about the clinical characteristics, predisposing factors and outcome of C fetus meningitis in adults. We report cases of C fetus meningitis in a nationwide cohort study of adult bacterial meningitis patients in the Netherlands and performed a review of the literature. Two patients with C fetus meningitis were identified from January 2006 through May 2015. The calculated annual incidence was 0.02 per million adults. Combined with the literature, we identified 22 patients with a median age of 48 years. An immunocompromised state was present in 16 patients (73%), mostly due to alcoholism (41%) and diabetes mellitus (27%). The source of infection was identified in 13 out of 19 patients (68%), consisting of regular contact with domestic animals in 5 and working on a farm in 4. Recurrent fever and illness was reported in 4 patients (18%), requiring prolonged antibiotic treatment. Two patients died (9%) and 3 survivors (15%) had neurological sequelae. C fetus is a rare cause of bacterial meningitis and is associated with an immunocompromised state. Based on the apparent slow clinical response seen in this limited number of cases, the authors of this study recommend a prolonged course of antimicrobial therapy when C fetus is identified as a causative agent of bacterial meningitis. Cases appeared to do best with carbapenem therapy. PMID:26937916

  15. Growth of non-Campylobacter, oxidase-positive bacteria on selective Campylobacter agar.

    OpenAIRE

    Moskowitz, L B; Chester, B

    1982-01-01

    A total of 67 oxidase-positive, gram-negative bacteria were tested for growth on selective Campylobacter agar (Blaser formulation, BBL Microbiology Systems, Cockeysville, Md.) at 42 degrees C under microaerophilic conditions. Although the growth of most of these bacteria was prevented, all strains of Achromobacter xylosoxidans, Pseudomonas aeruginosa, Pseudomonas putrefaciens, Pseudomonas alcaligenes, and Pseudomonas pseudoalcaligenes grew as well as Campylobacter fetus subsp. jejuni.

  16. Perspectives on deciphering mechanisms underlying plant heat stress response and thermotolerance

    Directory of Open Access Journals (Sweden)

    Kamila Lucia Bokszczanin

    2013-08-01

    Full Text Available Global warming is a major threat for agriculture and food safety and in many cases the negative effects are already apparent. The current challenge of basic and applied plant science is to decipher the molecular mechanisms of heat stress response and thermotolerance in detail and use this information to identify genotypes that will withstand unfavorable environmental conditions. Nowadays X-omics approaches complement the findings of previous targeted studies and highlight the complexity of heat stress response mechanisms giving information for so far unrecognized genes, proteins and metabolites as potential key players of thermotolerance. Even more, roles of epigenetic mechanisms and the involvement of small RNAs in thermotolerance are currently emerging and thus open new directions of yet unexplored areas of plant heat stress response. In parallel it is emerging that although the whole plant is vulnerable to heat, specific organs are particularly sensitive to elevated temperatures. This has redirected research from the vegetative to generative tissues. The sexual reproduction phase is considered as the most sensitive to heat and specifically pollen exhibits the highest sensitivity and frequently an elevation of the temperature just a few degrees above the optimum during pollen development can have detrimental effects for crop production. Compared to our knowledge on heat stress response of vegetative tissues, the information on pollen is still scarce. Nowadays, several techniques for high-throughput X-omics approaches provide major tools to explore the principles of pollen heat stress response and thermotolerance mechanisms in specific genotypes. The collection of such information will provide an excellent support for improvement of breeding programs to facilitate the development of tolerant cultivars. The review aims at describing the current knowledge of thermotolerance mechanisms and the technical advances which will foster new insights into

  17. Campylobacter Antimicrobial Resistance in Peru: A Ten-year Observational Study

    Science.gov (United States)

    2012-08-16

    Miller WG, Konkel ME: Differentiation of Campylobacter coli, Campylobacter jejuni, Campylobacter lari, and Campylobacter upsaliensis by a multiplex PCR ...van Pelt W, Wagenaar JA, Kuijper EJ: Inaccuracy of routine susceptibility tests for detection of erythromycin resistance of Campylobacter jejuni...available soon. Campylobacter antimicrobial resistance in Peru: a ten-year observational study BMC Infectious Diseases 2012, 12:193 doi:10.1186/1471-2334-12

  18. 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.

  19. Prevalence of Thermophilic Campylobacter species in carcasses ...

    African Journals Online (AJOL)

    Bernt Lindtjorn

    However, antimicrobial resistance to clinically important drugs used for treatment (especially macrolides and fluoroquinolones) is increasingly reported for campylobacters (9). There is growing scientific evidence that the use of antibiotics in food animals, particularly in developed countries, leads to the development of.

  20. 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

  1. Campylobacter enteritis among children in Dembia District ...

    African Journals Online (AJOL)

    Objective: To estimate the magnitude of Campylobacter enteritis in children below fifteen years of age. Design: A cross-sectional survey. Setting: Seven villages found in the outskirts of Kolla Diba town were covered. The town is located 35 kilometres away from Gondar teaching hospital. Participants: Stool specimens were ...

  2. CAMPYLOBACTER ENTERITIS AMONG CHILDREN IN DEMBIA ...

    African Journals Online (AJOL)

    hi-tech

    2000-12-12

    Dec 12, 2000 ... Objective: To estimate the magnitude of Campylobacter enteritis in children below fifteen years of age. Design: A cross-sectional survey. Setting: Seven villages found in the outskirts of Kolla Diba town were covered. The town is located 35 kilometres away from Gondar teaching hospital. Participants: Stool ...

  3. Molecular characterization of thermophilic Campylobacter species ...

    African Journals Online (AJOL)

    We identified two species of thermophilic Campylobacter in companion dogs in Jos. Majority of C. jejuni were isolated from mucoid faeces while mixed infections of the two species were more common among diarrhoeic dogs. Pet owners should observe strict hand hygiene especially after handling dogs or their faeces to ...

  4. Isolation and molecular characterization of Campylobacter coli

    African Journals Online (AJOL)

    Campylobacter coli is prevalent among trade pigs in Kafanchan, Nigeria and is distributed across four of the five states from which trade pigs were sourced. Adequate hand hygiene is recommended for farmers, traders and Veterinary professionals handling pigs to prevent the transmission of this zoonosis to humans.

  5. Campylobacter Antimicrobial Drug Resistance among Humans in ...

    African Journals Online (AJOL)

    Background: Though Campylobacter enteritis is a self-limiting disease, antimicrobial agents are recommended for extraintestinal infections and for treating immunocompromised persons. Erythromycin and ciprofloxacin are drugs of choice. The rate of resistance to these drugs is increasing in both developed and developing ...

  6. Prevalence of Campylobacter foetus and Trichomonas foetus ...

    African Journals Online (AJOL)

    Trichomoniasis and campylobacteriosis are diseases caused by Trichomonas foetus and Campylobacter foetus respectively. These diseases pose economic losses due to infertility and abortion. The aim of this retrospective study was to estimate the prevalence of C. foetus and T. foetus among southern African cattle.

  7. Campylobacter Spp. Epidemiology and Antimicrobial Susceptibility ...

    African Journals Online (AJOL)

    The isolation rate of Campylobacter was 2.3%,comprising of the following species C. jejuni (51.8%), C. coli (13.8%), and C. upsaliensis (3.5%). However, 30.9% of the isolates were unidentified. No resistant strain was found to gentamicin. The resistance to amoxicillin+clavulanic acid (3.4%) was lower than those ...

  8. 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

  9. Enhanced ethanol production from sugarcane juice by galactose adaptation of a newly isolated thermotolerant strain of Pichia kudriavzevii

    NARCIS (Netherlands)

    Dhaliwal, S.S.; Oberoi, H.S.; Sandhu, S.K.; Nanda, D.; Kumar, D.; Uppal, S.K.

    2011-01-01

    The thermotolerant yeast strain isolated from sugarcane juice through enrichment technique was identified as a strain of Pichiakudriavzevii (Issatchenkiaorientalis) through molecular characterization. The P. kudriavzevii cells adapted to galactose medium produced about 30% more ethanol from

  10. Detection of Campylobacter concisus and other Campylobacter species in colonic biopsies from adults with ulcerative colitis.

    Directory of Open Access Journals (Sweden)

    Indrani Mukhopadhya

    Full Text Available INTRODUCTION: The critical role of bacteria in the pathogenesis of ulcerative colitis (UC is well recognized, but an individual causative microorganism has not been singled out so far. Campylobacter concisus and other non-jejuni species of Campylobacter have been implicated as putative aetiological agents in inflammatory bowel disease in children, but such studies have not been addressed in adults. This study investigated the prevalence of Campylobacter species in colonic biopsy samples from adults with UC and healthy controls. METHODS: Adult patients who were undergoing diagnostic colonoscopy were recruited for the study, which included 69 patients with histologically proven UC and 65 healthy controls. DNA was extracted from the biopsy samples and subjected to Campylobacter genus specific and Campylobacter concisus specific polymerase chain reaction and sequencing. RESULTS: Detection of all Campylobacter DNA utilising genus specific primers was significantly higher in cases of UC, with a prevalence of 73.9% (51/69 compared to 23.1% (15/65 in controls (p = 0.0001. Nested PCR for C. concisus DNA was positive in 33.3% (23/69 of biopsy samples from subjects with UC, which was significantly higher than the prevalence rate of 10.8% (7/65 from controls (p = 0.0019. Sequencing of the remaining Campylobacter positive samples revealed that Campylobacter ureolyticus was positive in 21.7% (15/69 of samples from UC subjects as opposed to 3.1% (2/65 in controls (p = 0.0013. Mixed Campylobacter species were more common in UC patients, 20.3% (14/69 as compared to controls 4.6% (3/65 (p = 0.0084. CONCLUSION: The higher prevalence of Campylobacter genus and more specifically C. concisus and C. ureolyticus in biopsy samples from adults with UC suggests these genera of bacteria may be involved in the chronic inflammation that is characteristically seen in UC. To the best of our knowledge this is the first report of this association of C. concisus

  11. Detection of Campylobacter concisus and Other Campylobacter Species in Colonic Biopsies from Adults with Ulcerative Colitis

    Science.gov (United States)

    Mukhopadhya, Indrani; Thomson, John M.; Hansen, Richard; Berry, Susan H.; El-Omar, Emad M.; Hold, Georgina L.

    2011-01-01

    Introduction The critical role of bacteria in the pathogenesis of ulcerative colitis (UC) is well recognized, but an individual causative microorganism has not been singled out so far. Campylobacter concisus and other non-jejuni species of Campylobacter have been implicated as putative aetiological agents in inflammatory bowel disease in children, but such studies have not been addressed in adults. This study investigated the prevalence of Campylobacter species in colonic biopsy samples from adults with UC and healthy controls. Methods Adult patients who were undergoing diagnostic colonoscopy were recruited for the study, which included 69 patients with histologically proven UC and 65 healthy controls. DNA was extracted from the biopsy samples and subjected to Campylobacter genus specific and Campylobacter concisus specific polymerase chain reaction and sequencing. Results Detection of all Campylobacter DNA utilising genus specific primers was significantly higher in cases of UC, with a prevalence of 73.9% (51/69) compared to 23.1% (15/65) in controls (p = 0.0001). Nested PCR for C. concisus DNA was positive in 33.3% (23/69) of biopsy samples from subjects with UC, which was significantly higher than the prevalence rate of 10.8% (7/65) from controls (p = 0.0019). Sequencing of the remaining Campylobacter positive samples revealed that Campylobacter ureolyticus was positive in 21.7% (15/69) of samples from UC subjects as opposed to 3.1% (2/65) in controls (p = 0.0013). Mixed Campylobacter species were more common in UC patients, 20.3% (14/69) as compared to controls 4.6% (3/65) (p = 0.0084). Conclusion The higher prevalence of Campylobacter genus and more specifically C. concisus and C. ureolyticus in biopsy samples from adults with UC suggests these genera of bacteria may be involved in the chronic inflammation that is characteristically seen in UC. To the best of our knowledge this is the first report of this association of C. concisus and C

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

    Directory of Open Access Journals (Sweden)

    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.

  13. 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.

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

    DEFF Research Database (Denmark)

    Hald, Birthe; Pedersen, Karl; Wainø, Michael

    2004-01-01

    The Campylobacter excretion patterns of 26 domestic pet dogs were described in a longitudinal study. The dogs entered the study between 3 and 8 months of age and were monitored until 2 years of age. They were tested monthly for Campylobacter carriage in stool samples that were cultured...... on the Campylobacter-selective media CAT and modified CCDA agar at 37 and 42 C. This study comprised 366 fecal swab samples, of which 278 (76.2%) were found to be Campylobacter positive, with the following distribution of species: 75.0% Campylobacter upsaliensis, 19.4% Campylobacter jejuni, 2.1% Campylobacter lari, 0.......7% Campylobacter coli, and 2.8% Campylobacter spp. Isolates were typed by pulsed-field gel electrophoresis (PFGE) to elucidate the strain excretion pattern. All study dogs excreted Campylobacter spp. during the study period. At 3 months of age, 60% of the dogs carried Campylobacter, increasing to nearly 100...

  15. 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.

  16. Campylobacter in the environment: A major threat to public health

    Directory of Open Access Journals (Sweden)

    Hussein Hasan Abulreesh

    2017-06-01

    Full Text Available Epidemiological data suggest that Campylobacter remains a worldwide leading cause of gastrointestinal infections. Improperly prepared meat products, unpasteurized milk as well as non chlorinated drinking water were shown to be the main sources of campylobacteriosis. The Campylobacter survival mechanism in various environments facilitated the transmission of Campylobacter-associated infections; however the exact mode of transmission remains to be elucidated. This review aims to summarize recent insights on the incidence and survival of Campylobacter in the environment. Besides, methods of detection and risk assessment for public health safety are also addressed.

  17. 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.

  18. 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....

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

    Directory of Open Access Journals (Sweden)

    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.

  20. Non-stochastic sampling error in quantal analyses for Campylobacter species on poultry products.

    Science.gov (United States)

    Irwin, Peter; Reed, Sue; Brewster, Jeffrey; Nguyen, Ly; He, Yiping

    2013-03-01

    Using primers and fluorescent probes specific for the most common food-borne Campylobacter species (Campylobacter jejuni and Campylobacter coli), we developed a multiplex, most probable number (MPN) assay using quantitative PCR (qPCR) as the determinant for binomial detection: i.e., number of p positive pathogen growth responses out of n = 6 observations each of 4 mL (V) per dilution. Working with media washes of thrice frozen-thawed chicken pieces which had been spiked with known levels of C. jejuni and C. coli, we found that about 20% of the experiments had a significant amount of error in the form of either greater than 25% MPN calculation error (Δε) and/or a low apparent recovery rate (R less than 1 = MPN observed ÷ CFU spiked). Assuming such errors were exacerbated by an excessively small n, we examined computer-generated MPN enumeration data from the standpoint of stochastic sampling error (Δ) and found that such binomial-based assays behaved identically to Poisson-based methods (e.g., counting data) except that fewer technical replicates (n) appeared to be required for the same number of cells per test volume (μ). This result implies that the qPCR detection-based MPN protocol discussed herein should accurately enumerate a test population with a μ ≥ 1 using n = 6 observations per dilution. For our protocol, this equates to ≥ 8 cells per 400-500 g of sampled product. Based on this analysis, the error rate we saw in spiked experiments (where μ > 1) implied a non-stochastic source. In other experiments we present evidence that this source was, at least in part, related to the cell concentration step (i.e., centrifugation). We also demonstrate that the error rate lessened (from ~38% to ~13%) at lower Campylobacter levels (μ ≤ 40) as would most likely exist in nature. Using this protocol, we were able to quantify 14 to 1,226 MPN per 450 g of naturally contaminated chicken for skinless pieces and 11 to 244 MPN per 450 g for wings, breasts, legs, and

  1. The detection of hipO gene by real-time PCR in thermophilic Campylobacter spp. with very weak and negative reaction of hippurate hydrolysis.

    Science.gov (United States)

    Caner, Vildan; Cokal, Yavuz; Cetin, Cengiz; Sen, Aysin; Karagenc, Nedim

    2008-11-01

    A total of 190 Campylobacter spp. isolates, of which 34 gave the result of very weak activity, and 156 gave the negative activity in the test for hippurate hydrolysis were characterized. The genomic DNA was isolated from a fresh culture of each isolate and the real-time PCR, targeting the hipO gene, was used to confirm the species distribution of Campylobacter isolates. The hipO gene was detected in 17 isolates (11%) within the total of 156 negative isolates for hippurate hydrolysis. Out of 34 isolates with very weak activity, 19 isolates (56%) were also found to be positive for hipO gene and characterized as C. jejuni. The real-time PCR assay used in this study could be employed for more accurate diagnosis of Campylobacter infections at species level after the biochemical characterization based on hippuricase activity of the isolates. This could also provide important data for the epidemiology of infections associated with these zoonotic pathogens.

  2. Genetic manipulation of Bacillus methanolicus, a gram-positive, thermotolerant methylotroph.

    OpenAIRE

    Cue, D; Lam, H; Dillingham, R L; Hanson, R S; Flickinger, M C

    1997-01-01

    We report the fist genetic transformation system, shuttle vectors, and integrative vectors for the thermotolerant, methylotrophic bacterium Bacillus methanolicus. By using a polyethylene glycol-mediated transformation procedure, we have successfully transformed B. methanolicus with both integrative and multicopy plasmids. For plasmids with a single BmeTI recognition site, dam methylation of plasmid DNA (in vivo or in vitro) was found to enhance transformation efficiency from 7- to 11-fold. Tw...

  3. Salinity modulates thermotolerance, energy metabolism and stress response in amphipods Gammarus lacustris

    OpenAIRE

    Vereshchagina, Kseniya P.; Lubyaga, Yulia A.; Shatilina, Zhanna; Bedulina, Daria; Gurkov, Anton; Axenov-Gribanov, Denis V.; Baduev, Boris; Kondrateva, Elizaveta S.; Gubanov, Mikhail; Zadereev, Egor; Sokolova, Inna; Timofeyev, Maxim

    2016-01-01

    Temperature and salinity are important abiotic factors for aquatic invertebrates. We investigated the influence of different salinity regimes on thermotolerance, energy metabolism and cellular stress defense mechanisms in amphipods Gammarus lacustris Sars from two populations. We exposed amphipods to different thermal scenarios and determined their survival as well as activity of major antioxidant enzymes (peroxidase, catalase, glutathione S-transferase) and parameters of energy metabolism (c...

  4. Consortium inoculum of five thermo-tolerant phosphate solubilizing Actinomycetes for multipurpose biofertilizer preparation.

    Science.gov (United States)

    Nandimath, Arusha P; Karad, Dilip D; Gupta, Shantikumar G; Kharat, Arun S

    2017-10-01

    Alkaline pH of the soil facilitates the conversion of phosphate present in phosphate fertilizer applied in the field to insoluble phosphate which is not available to plants. Problem of soluble phosphate deficiency arises, primarily due to needless use of phosphate fertilizer. We sought to biofertilizer with the thermo-tolerant phosphate solubilizing actinomycetes consortium that could convert insoluble phosphate to soluble phosphate at wider temperature range. In the present investigation consortium of five thermo-tolerant phosphate solubilizing actinomycetes was applied for preparation of inoculum to produce multipurpose bio-fertilizer. Phosphates solubilizing thermo-tolerant 32 actinomycetes strains were processed for identification with the use of PIBWIN software and were screened for phosphate solubilizing activity. Amongst these five actinomycetes were selected on the basis of their ability to produce cellulase, chitinase, pectinase, protease, lipase, amylase and phosphate solubilizing enzymes. Ability to produce these enzymes at 28°C and 50°C were examined. Biofertilizer was prepared by using agricultural waste as a raw material. While preparation of bio-fertilizer the pH decreased from 7.5 to 4.3 and temperature increased up to 74°C maximum at the end of 4 th week and in subsequent week it started to decline gradually till it reached around 50°C, which was found to be stable up to eighth week. This thermo-tolerant actinomycetes consortium released soluble phosphate of up to 46.7 μg ml -1 . As the mesophilic organisms die out at high temperature of composting hence thormo-tolerant actinomycetes would be the better substitute for preparation of phosphate solubilizing bio-fertilizer with added potential to degrade complex macromolecules in composting.

  5. Comparison of thermotolerant coliforms and Escherichia coli densities in freshwater bodies.

    Science.gov (United States)

    Hachich, Elayse M; Di Bari, Marisa; Christ, Ana Paula G; Lamparelli, Cláudia C; Ramos, Solange S; Sato, Maria Inês Z

    2012-04-01

    Fecal bacterial indicator analyses have been widely used for monitoring the water quality. This study was designed to determine the ratio between the density of Escherichia coli and other Thermotolerant Coliforms (TtC) bacteria from freshwater samples collected for a two-year period of monitoring. TtC were enumerated by membrane filtration on mFC agar. E. coli enumeration was done by two methods: TtC colonies identified in mFC were inoculated in EC-MUG or water samples were filtered and inoculated in modified mTEC agar media, and both methods were compared for quantitative recovery of E. coli. The results pointed out a mean percentage of E. coli among other thermotolerant coliforms (E. coli/TtC ratio) of 84.3% in mFC media. Taking these results into account, a mandatory standard of 1000 thermotolerant coliforms would correspond to 800 E. coli and the adoption of these E. coli based standards will represent a major improvement for the monitoring of freshwater quality.

  6. Trampling Impacts on Thermotolerant Vegetation of Geothermal Areas in New Zealand

    Science.gov (United States)

    Burns, Bruce R.; Ward, Jonet; Downs, Theresa M.

    2013-12-01

    Geothermal features such as geysers, mud pools, sinter terraces, fumaroles, hot springs, and steaming ground are natural attractions often visited by tourists. Visitation rates for such areas in the Taupo Volcanic Zone of New Zealand are in the order of hundreds of thousands annually. These areas are also habitat for rare and specialized plant and microbial communities that live in the steam-heated soils of unusual chemical composition. We evaluated historical and current trampling impacts of tourists on the thermotolerant vegetation of the Waimangu and Waiotapu geothermal areas near Rotorua, and compared the results to experimental trampling at a third site (Taheke) not used by tourists. Historical tourism has removed vegetation and soil from around key features, and remaining subsoil is compacted into an impervious pavement on which vegetation recolonization is unlikely in the short term. Social tracks made by tourists were present at both tourist sites often leading them onto hotter soils than constructed tracks. Vegetation height and cover were lower on and adjacent to social tracks than further from them. Thermotolerant vegetation showed extremely low resistance to experimental trampling. This confirms and extends previous research that also shows that thallophytes and woody shrubs, life forms that dominate in thermotolerant vegetation, are vulnerable to trampling damage. Preservation of these vulnerable ecosystems must ensure that tourist traffic is confined to existing tracks or boardwalks, and active restoration of impacted sites may be warranted.

  7. Trampling impacts on thermotolerant vegetation of geothermal areas in New Zealand.

    Science.gov (United States)

    Burns, Bruce R; Ward, Jonet; Downs, Theresa M

    2013-12-01

    Geothermal features such as geysers, mud pools, sinter terraces, fumaroles, hot springs, and steaming ground are natural attractions often visited by tourists. Visitation rates for such areas in the Taupo Volcanic Zone of New Zealand are in the order of hundreds of thousands annually. These areas are also habitat for rare and specialized plant and microbial communities that live in the steam-heated soils of unusual chemical composition. We evaluated historical and current trampling impacts of tourists on the thermotolerant vegetation of the Waimangu and Waiotapu geothermal areas near Rotorua, and compared the results to experimental trampling at a third site (Taheke) not used by tourists. Historical tourism has removed vegetation and soil from around key features, and remaining subsoil is compacted into an impervious pavement on which vegetation recolonization is unlikely in the short term. Social tracks made by tourists were present at both tourist sites often leading them onto hotter soils than constructed tracks. Vegetation height and cover were lower on and adjacent to social tracks than further from them. Thermotolerant vegetation showed extremely low resistance to experimental trampling. This confirms and extends previous research that also shows that thallophytes and woody shrubs, life forms that dominate in thermotolerant vegetation, are vulnerable to trampling damage. Preservation of these vulnerable ecosystems must ensure that tourist traffic is confined to existing tracks or boardwalks, and active restoration of impacted sites may be warranted.

  8. Screening of thermotolerant and thermophilic fungi aiming β-xylosidase and arabinanase production

    Science.gov (United States)

    Benassi, Vivian Machado; de Lucas, Rosymar Coutinho; Jorge, João Atílio; Polizeli, Maria de Lourdes Teixeira de Moraes

    2014-01-01

    Plant cell wall is mainly composed by cellulose, hemicellulose and lignin. The heterogeneous structure and composition of the hemicellulose are key impediments to its depolymerization and subsequent use in fermentation processes. Thus, this study aimed to perform a screening of thermophilic and thermotolerant filamentous fungi collected from different regions of the São Paulo state, and analyze the production of β-xylosidase and arabinanase at different temperatures. These enzymes are important to cell wall degradation and synthesis of end products as xylose and arabinose, respectively, which are significant sugars to fermentation and ethanol production. A total of 12 fungal species were analyzed and 9 of them grew at 45 °C, suggesting a thermophilic or thermotolerant character. Additionally Aspergillus thermomutatus anamorph of Neosartorya and A. parasiticus grew at 50 °C. Aspergillus niger and Aspergillus thermomutatus were the filamentous fungi with the most expressive production of β-xylosidase and arabinanase, respectively. In general for most of the tested microorganisms, β-xylosidase and arabinanase activities from mycelial extract (intracellular form) were higher in cultures grown at high temperatures (35–40 °C), while the correspondent extracellular activities were favorably secreted from cultures at 30 °C. This study contributes to catalogue isolated fungi of the state of São Paulo, and these findings could be promising sources for thermophilic and thermotolerant microorganisms, which are industrially important due to their enzymes. PMID:25763055

  9. Microbiological aspects of Helicobacter pylori (Campylobacter pylori).

    Science.gov (United States)

    Goodwin, C S; Armstrong, J A

    1990-01-01

    The human gastric pathogen Campylobacter pylori has recently been reclassified as Helicobacter pylori, and a related spiral bacterium found in the stomach of ferrets has been designated Helicobacter mustelae. The general microbiological features of Helicobacter pylori are delineated here, with details of phenotypic differences between Helicobacter pylori and Helicobacter mustelae; comparisons are made with Wolinella succinogenes and Campylobacter jejuni. The Helicobacter organisms possess an external glycocalyx which can be visualised by electron microscopy, and which may be involved in bacterial adherence. The finding of soluble and cell-associated haemagglutinins of Helicobacter pylori is reported. Detection of Helicobacter pylori in clinical specimens, susceptibility of the organism to antibacterial agents, and other aspects of practical and clinical significance are briefly reviewed.

  10. Infeções alimentares por Campylobacter

    OpenAIRE

    Neto, Carlota Duarte Castro

    2017-01-01

    Dissertação para obtenção do grau de Mestre no Instituto Superior de Ciências da Saúde Egas Moniz Após o isolamento bem-sucedido em fezes na década de setenta do século passado (1972), Campylobacter tornou-se rapidamente a espécie bacteriana mais comumente associada a doenças diarreicas em todo o mundo. A infeção por Campylobacter provoca um espectro de doenças, incluindo enterite aguda, infeções extra-gastrointestinais e complicações pós-infeciosas. A doença gastrointestinal autolimita...

  11. 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.

  12. 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.

  13. 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,

  14. Thermotolerance and responses to short duration heat stress in tropical and temperate species

    Science.gov (United States)

    Marias, D.; Meinzer, F. C.; Still, C. J.

    2017-12-01

    Temperature and heat waves are predicted to increase throughout the 21st century in both tropical and temperate regions. Tropical species are vulnerable to heat stress because of the higher radiation load and the narrower distribution of temperatures typically experienced compared to extratropical species. Germinant seedlings are also vulnerable to heat stress because they inhabit the boundary layer close to the soil surface where intense heating occurs. We quantified the effect of leaf age and heat stress duration (45 min, 90 min) on leaf thermotolerance and whole plant physiological responses to heat stress in Coffea arabica (COAR) saplings. We also evaluated leaf thermotolerance and whole plant responses to heat stress of seedlings in two populations each of Pinus ponderosa (PIPO) and Pseudotsuga menziesii (PSME) from contrasting climates. Thermotolerance of detached leaves/needles was evaluated using chlorophyll fluorescence (FV/FM, FO) and electrolyte leakage. After exposure of whole plants to a simulated heat wave in a growth chamber, we monitored FV/FM, photosynthesis (A), stomatal conductance (gs), non-structural carbohydrates (NSCs), and carbon isotope ratios (δ13C). In COAR, thermotolerance and rate of recovery increased with leaf age. Following heat treatment, reductions in A and gs led to reduced intrinsic water use efficiency (iWUE) and increased leaf temperatures. NSC results suggested that starch was converted to sugars for recovery from heat stress and phloem transport was inhibited. Plants failed to flower in both heat stress duration treatments. In PIPO and PSME, heat treatment induced significant reductions in FV/FM and A. NSC results suggested that starch was converted to glucose + fructose to aid recovery from heat-induced damage. Populations from drier sites had greater δ13C values than those from wetter sites, consistent with higher iWUE of populations from drier climates. Thermotolerance and heat stress responses appeared to be

  15. Prevalence of Campylobacter spp. in skinless, boneless retail broiler meat from 2005 through 2011 in Alabama, USA

    Directory of Open Access Journals (Sweden)

    Williams Aretha

    2012-08-01

    Full Text Available Abstract Background The prevalence of Campylobacter spp. in 755 skinless, boneless retail broiler meat samples (breast, tenderloins and thighs collected from food stores in Alabama, USA, from 2005 through 2011 was examined. Campylobacter spp. were isolated using enrichment and plate media. Isolates were identified with multiplex PCR assays and typed with pulsed field gel electrophoresis (PFGE. Data were analyzed by nominal variables (brand, plant, product, season, state and store that may affect the prevalence of these bacteria. Results The average prevalence of Campylobacter spp. in retail broiler meat for these years was 41%, with no statistical differences in the prevalence by year (P > 0.05. Seasons did not affect the prevalence of C. jejuni but statistically affected the prevalence of C. coli (P P P C. coli and C. jejuni had an average prevalence of 28% and 66%, respectively. The prevalence of C. coli varied by brand, plant, season, state, store and year, while the prevalence of C. jejuni varied by brand, product, state and store. Tenderloins had a lower prevalence of Campylobacter spp. than breasts and thighs (P P > 0.05 were observed in the prevalence of C. jejuni by season, the lowest prevalence of C. coli was recorded from October through March. A large diversity of PFGE profiles was found for C. jejuni, with some profiles from the same processing plants reappearing throughout the years. Conclusions The prevalence of Campylobacter spp. did not change during the seven years of the study; however, it did change when analyzed by brand, product and state. Seasons did not affect the prevalence of C. jejuni, but they did affect the prevalence of C. coli. Larger PFGE databases are needed to assess the temporal reoccurrence of PFGE profiles to help predict the risk associated with each profile.

  16. Novel anti-campylobacter compounds identified using high throughput screening of a pre-selected enriched small molecules library

    Directory of Open Access Journals (Sweden)

    Anand eKumar

    2016-04-01

    Full Text Available Campylobacter is a leading cause of foodborne bacterial gastroenteritis worldwide and infections can be fatal. The emergence of antibiotic-resistant Campylobacter spp. necessitates the development of new antimicrobials. We identified novel anti-Campylobacter small molecule inhibitors using a high throughput growth inhibition assay. To expedite screening, we made use of a bioactive library of 4,182 compounds that we have previously shown to be active against diverse microbes. Screening for growth inhibition of Campylobacter jejuni, identified 781 compounds that were either bactericidal or bacteriostatic at a concentration of 200 µM. Seventy nine of the bactericidal compounds were prioritized for secondary screening based on their physico-chemical properties. Based on the minimum inhibitory concentration against a diverse range of C. jejuni and a lack of effect on gut microbes, we selected 12 compounds. No resistance was observed to any of these 12 lead compounds when C. jejuni was cultured with lethal or sub-lethal concentrations suggesting that C. jejuni is less likely to develop resistance to these compounds. Top 12 compounds also possessed low cytotoxicity to human intestinal epithelial cells (Caco-2 cells and no hemolytic activity against sheep red blood cells. Next, these 12 compounds were evaluated for ability to clear C. jejuni in vitro. A total of 10 compounds had an anti-C. jejuni effect in Caco-2 cells with some effective even at 25 µM concentrations. These novel 12 compounds belong to five established antimicrobial chemical classes; piperazines, aryl amines, piperidines, sulfonamide and pyridazinone. Exploitation of analogues of these chemical classes may provide Campylobacter specific drugs that can be applied in both human and animal medicine.

  17. 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

  18. Campylobacter jejuni seroepidemiology in native chicken

    Directory of Open Access Journals (Sweden)

    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.

  19. Molecular Diagnosis of Shigella, Salmonella and Campylobacter by ...

    African Journals Online (AJOL)

    infections, enteric bacteria such as Campylobacter, Salmonella and Shigella are impor- tant because of the frequency and severity of the symptoms they could cause. Thus,. Campylobacter is currently considered the leading cause of intestinal bacterial infec- tions in humans worldwide with an increasing incidence in ...

  20. Foodborne disease prevention and broiler chickens with reduced Campylobacter infection

    DEFF Research Database (Denmark)

    Bahrndorff, Simon; Rangstrup-Christensen, Lena; Nordentoft, Steen

    2013-01-01

    Studies have suggested that flies play a linking role in the epidemiology of Campylobacter spp. in broiler chickens and that fly screens can reduce the prevalence of Campylobacter spp. We examined the year-round and long-term effects of fly screens in 10 broiler chicken houses (99 flocks...... broiler chicken flocks....

  1. Campylobacter: animal reservoirs, human infections, and options for control

    NARCIS (Netherlands)

    Wagenaar, Jaap; Newell, D.G.; Kalupahana, R.S.; Mughini Gras, Lapo

    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

  2. Campylobacter infections in fattening pigs; excretion pattern and genetic diversity

    NARCIS (Netherlands)

    Weijtens, M.J.B.M.; Reinders, R.D.; Urlings, H.A.P.; Plas, van der J.

    1999-01-01

    The excretion of campylobacter by eight individually housed fattening pigs was monitored during 15 weeks. Rectal faeces samples were collected six times from these pigs and twice from their mothers (seven sows). Campylobacter was cultured from these samples on Preston medium. In some pigs, samples

  3. Host-pathogen interactions in Campylobacter infections: the host perspective

    NARCIS (Netherlands)

    Janssen, R.; Krogfelt, K.A.; Cawthraw, S.A.; Pelt, van W.; Wagenaar, J.A.; Owen, R.J.

    2008-01-01

    Campylobacter is a major cause of acute bacterial diarrhea in humans worldwide. This study was aimed at summarizing the current understanding of host mechanisms involved in the defense against Campylobacter by evaluating data available from three sources: (i) epidemiological observations, (ii)

  4. Campylobacter infections in fattening pigs; Excretion pattern and genetic diversity

    NARCIS (Netherlands)

    Weijtens, M.J.B.M.; Reinders, R.D.; Urlings, H.A.P.; Plas, J. van der

    1999-01-01

    The excretion of campylobacter by eight individually housed fattening pigs was monitored during 15 weeks. Rectal faeces samples were collected six times from these pigs and twice from their mothers (seven sows). Campylobacter was cultured from these samples on Preston medium. In some pigs, samples

  5. 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...

  6. 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

  7. Campylobacter pylori as possible factor in peptic ulcer recurrence

    NARCIS (Netherlands)

    Rauws, E. A.

    1989-01-01

    The author reviews the literature up to 1988 about the close association of Campylobacter pylori with chronic active gastritis, duodenitis and peptic ulcer disease. No firm data however demonstrate that Campylobacter pylori causes duodenal ulcer but long term eradication of this bacterium prevents

  8. 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...

  9. Effect of Noradrenaline on the Virulence Properties of Campylobacter Species

    Directory of Open Access Journals (Sweden)

    Sree V. Aroori

    2014-01-01

    Full Text Available Campylobacter species cause a spectrum of illnesses in humans. The type of illness and the outcome is dependent on the virulence of the infecting pathogen strain and host immune status. Acute stress can seriously compromise host immunity and increase susceptibility to infection. Noradrenaline (NA is a stress hormone. Several studies have shown that it stimulated growth and increased the pathogenicity of organisms including E. coli and Campylobacter jejuni. However, the effect of NA on other Campylobacter species is unknown. We have examined the effect of NA on growth rate, motility, invasion of T84 epithelial cells, and colonisation of chickens by diverse Campylobacter species. Campylobacter cultures grown with NA had reduced lag phases, increased growth rates, and higher final optical densities than controls. The motility of Campylobacter was also significantly increased in the presence of noradrenaline. Some of the Campylobacter strains tested also showed increased invasion of T84 epithelial cells, greater breakdown of tight junctions, and an enhanced potential to colonise chickens. Our results show that noradrenaline-induced enhancement of virulence of Campylobacter can influence the outcome of infection.

  10. 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

  11. Campylobacter spp among Children with acute diarrhea attending ...

    African Journals Online (AJOL)

    Isolation rate in developing countries is between 5-35%. This study aimed at finding prevalence of children with campylobacter infection among children with acute diarrhea attending Mulago hospital. Objective: The objective was to establish the proportion of children infected with Campylobacter spp among children with ...

  12. Preventing Campylobacter at the Source: Why Is It So Difficult?

    NARCIS (Netherlands)

    Wagenaar, J.A.; French, N.P.; Havelaar, A.H.

    2013-01-01

    Campylobacteriosis in humans, caused by Campylobacter jejuni and Campylobacter coli, is the most common recognized bacterial zoonosis in the European Union and the United States. The acute phase is characterized by gastrointestinal symptoms. The long-term sequelae (Guillain-Barre syndrome, reactive

  13. 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

  14. Enzyme assays.

    Science.gov (United States)

    Brodelius, P E

    1991-02-01

    The past year or so has seen the development of new enzyme assays, as well as the improvement of existing ones. Assays are becoming more rapid and sensitive as a result of modifications such as amplification of the enzyme product(s). Recombinant DNA technology is now being recognized as a particularly useful tool in the search for improved assay systems.

  15. 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

  16. Restaurant Cooking Trends and Increased Risk for Campylobacter Infection.

    Science.gov (United States)

    Jones, Anna K; Rigby, Dan; Burton, Michael; Millman, Caroline; Williams, Nicola J; Jones, Trevor R; Wigley, Paul; O'Brien, Sarah J; Cross, Paul

    2016-07-01

    In the United Kingdom, outbreaks of Campylobacter infection are increasingly attributed to undercooked chicken livers, yet many recipes, including those of top chefs, advocate short cooking times and serving livers pink. During 2015, we studied preferences of chefs and the public in the United Kingdom and investigated the link between liver rareness and survival of Campylobacter. We used photographs to assess chefs' ability to identify chicken livers meeting safe cooking guidelines. To investigate the microbiological safety of livers chefs preferred to serve, we modeled Campylobacter survival in infected chicken livers cooked to various temperatures. Most chefs correctly identified safely cooked livers but overestimated the public's preference for rareness and thus preferred to serve them more rare. We estimated that 19%-52% of livers served commercially in the United Kingdom fail to reach 70°C and that predicted Campylobacter survival rates are 48%-98%. These findings indicate that cooking trends are linked to increasing Campylobacter infections.

  17. Antimicrobial resistance in campylobacter: susceptibility testing methods and resistance trends.

    Science.gov (United States)

    Ge, Beilei; Wang, Fei; Sjölund-Karlsson, Maria; McDermott, Patrick F

    2013-10-01

    Most Campylobacter infections are self-limiting but antimicrobial treatment (e.g., macrolides, fluoroquinolones) is necessary in severe or prolonged cases. Susceptibility testing continues to play a critical role in guiding therapy and epidemiological monitoring of resistance. The methods of choice for Campylobacter recommended by the Clinical and Laboratory Standards Institute (CLSI) are agar dilution and broth microdilution, while a disk diffusion method was recently standardized by the European Committee on Antimicrobial Susceptibility Testing (EUCAST). Macrolides, quinolones, and tetracyclines are among the common antimicrobials recommended for testing. Molecular determination of Campylobacter resistance via DNA sequencing or PCR-based methods has been performed. High levels of resistance to tetracycline and ciprofloxacin are frequently reported by many national surveillance programs, but resistance to erythromycin and gentamicin in Campylobacter jejuni remains low. Nonetheless, variations in susceptibility observed over time underscore the need for continued public health monitoring of Campylobacter resistance from humans, animals, and food. Published by Elsevier B.V.

  18. Strategies for the inclusion of an internal amplification control in conventional and real time PCR detection of Campylobacter spp. in chicken fecal samples

    DEFF Research Database (Denmark)

    Lund, Marianne; Madsen, Mogens

    2006-01-01

    To illustrate important issues in optimization of a PCR assay with an internal control four different primer combinations for conventional PCR, two non-competitive and two competitive set-ups for real time PCR were used for detection of Campylobacter spp. in chicken faecal samples. In the convent......To illustrate important issues in optimization of a PCR assay with an internal control four different primer combinations for conventional PCR, two non-competitive and two competitive set-ups for real time PCR were used for detection of Campylobacter spp. in chicken faecal samples....... In the conventional PCR assays the internal control was genomic DNA from Yersinia ruckeri, which is not found in chicken faeces. This internal control was also used in one of the set LIPS in real time PCR. In the three other set-ups different DNA fragments of 109 bp length prepared from two oligos of each 66 bp...

  19. 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

  20. Detection of Campylobacter Bacteria in Air Samples for Continuous Real-Time Monitoring of Campylobacter Colonization in Broiler Flocks

    DEFF Research Database (Denmark)

    Olsen, Katja Nyholm; Lund, Marianne; Skov, J.

    2009-01-01

    Improved monitoring tools are important for the control of Campylobacter bacteria in broiler production. In this study, we compare the sensitivities of detection of Campylobacter by PCR with feces, dust, and air samples during the lifetimes of broilers in two poultry houses and conclude that the ...

  1. Superior thermotolerance of Saccharomyces cerevisiae for efficient bioethanol fermentation can be achieved by overexpression of RSP5 ubiquitin ligase.

    Science.gov (United States)

    Shahsavarani, Hosein; Sugiyama, Minetaka; Kaneko, Yoshinobu; Chuenchit, Boonchird; Harashima, Satoshi

    2012-01-01

    The simultaneous saccharification and fermentation process requires thermo-tolerant yeast to facilitate the enzymatic hydrolysis of cellulose. In this paper, we describe a Htg+ strain that exhibits confluent growth at high temperature (41 °C) and resistance to heat shock, ethanol, osmotic, oxidative and DNA damage stresses. HTG6, one of the six genes responsible for the thermotolerant phenotype was identified to be the gene RSP5 encoding a ubiquitin ligase. The RSP5 allele of the Htg+ strain, designated RSP5-C, possessed five, one and two base changes in the promoter, open reading frame and terminator region, respectively. The base changes in the promoter region of the RSP5-C allele were found to be responsible for the thermotolerant phenotype by strongly increasing transcription of the RSP5 gene and consequently causing a rise in the ubiquitination of cell proteins. Overexpression of the RSP5-BY allele present in the htg6 host strain (Htg-) conferred thermotolerance at 41°C, to this strain as in the case of RSP5-C allele. We also discovered that an Htg+ strain overexpressing the RSP5-C allele exhibits a more robust Htg+ phenotype against higher temperature (43 °C). The data presented here also suggest that overexpression of RSP5 could be applied to raise the upper limit of thermotolerance in S. cerevisiae strain used for industrial bioethanol production. Copyright © 2011 Elsevier Inc. All rights reserved.

  2. Cost-effective optimization of real-time PCR based detection of Campylobacter and Salmonella with inhibitor tolerant DNA polymerases

    DEFF Research Database (Denmark)

    Fachmann, Mette Sofie Rousing; Josefsen, Mathilde Hasseldam; Hoorfar, Jeffrey

    2015-01-01

    bacterial cells in two validated real-time PCR assays for Campylobacter and Salmonella. The five best performing (based on: limit of detection (LOD), maximum fluorescence, shape of amplification curves, and amplification efficiency) were subsequently applied to meat and fecal samples. The VeriQuest q...... (LOD=103 -106 CFU ml-1 /not detected) with fecal samples. CONCLUSIONS: Applying the VeriQuest qPCR master mix in the two tested real-time PCR assays could allow for simpler sample preparation and thus a reduction in cost. SIGNIFICANCE AND IMPACT OF STUDY: This work exemplifies a cost-effective strategy...

  3. 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.

  4. A multiplex real-time PCR for the detection and differentiation of Campylobacter phages.

    Directory of Open Access Journals (Sweden)

    Claudia Jäckel

    Full Text Available Campylobacter jejuni and C. coli are important food-borne pathogens that are widespread in animal husbandry. To combat Campylobacter along the food chain, the application of lytic phages has been shown to be a promising tool. Campylobacter phages are currently classified into three groups, of which group II and group III phages are the most common. Members of each group are closely related, whereas the two groups share only little DNA similarity. Moreover, while group III phages are specific for C. jejuni, group II phages additionally infect C. coli. Phage cocktails intended to be used for applications should be composed of various phages that differ in their host range and growth kinetics. The isolation of phages is generally performed by plaque assays. This approach has the limitation that phages are merely identified by their lytic activity on certain indicator strains and that relatively high numbers of phages must be present in a tested sample. Therefore, a more sensitive molecular detection system would be beneficial, which allows a pre-screening of samples and the quick detection and discrimination of group II and group III phages. New phages can then be isolated by use of indicator strains that may be different from those typically applied. On the basis of available Campylobacter phage genome sequences, we developed a multiplex PCR system for group II and group III phages selecting the tail tube gene and the gene for the base plate wedge, respectively, as target. Phages of both groups could be identified with primers deduced from the putative tail fiber gene. Efficient release of phage DNA from capsids was achieved by an extended heat treatment or digestion of phage particles with proteinase K/SDS yielding a detection limit of 1 pfu/ml. Individual detection of group II phages, group III phages and of both groups was studied with artificially contaminated chicken skin. To recover phages that had strongly adhered to the skin, stomaching

  5. A multiplex real-time PCR for the detection and differentiation of Campylobacter phages.

    Science.gov (United States)

    Jäckel, Claudia; Hammerl, Jens A; Rau, Jörg; Hertwig, Stefan

    2017-01-01

    Campylobacter jejuni and C. coli are important food-borne pathogens that are widespread in animal husbandry. To combat Campylobacter along the food chain, the application of lytic phages has been shown to be a promising tool. Campylobacter phages are currently classified into three groups, of which group II and group III phages are the most common. Members of each group are closely related, whereas the two groups share only little DNA similarity. Moreover, while group III phages are specific for C. jejuni, group II phages additionally infect C. coli. Phage cocktails intended to be used for applications should be composed of various phages that differ in their host range and growth kinetics. The isolation of phages is generally performed by plaque assays. This approach has the limitation that phages are merely identified by their lytic activity on certain indicator strains and that relatively high numbers of phages must be present in a tested sample. Therefore, a more sensitive molecular detection system would be beneficial, which allows a pre-screening of samples and the quick detection and discrimination of group II and group III phages. New phages can then be isolated by use of indicator strains that may be different from those typically applied. On the basis of available Campylobacter phage genome sequences, we developed a multiplex PCR system for group II and group III phages selecting the tail tube gene and the gene for the base plate wedge, respectively, as target. Phages of both groups could be identified with primers deduced from the putative tail fiber gene. Efficient release of phage DNA from capsids was achieved by an extended heat treatment or digestion of phage particles with proteinase K/SDS yielding a detection limit of 1 pfu/ml. Individual detection of group II phages, group III phages and of both groups was studied with artificially contaminated chicken skin. To recover phages that had strongly adhered to the skin, stomaching was the most efficient

  6. Molecular and functional characterization of two pyruvate decarboxylase genes, PDC1 and PDC5, in the thermotolerant yeast Kluyveromyces marxianus.

    Science.gov (United States)

    Choo, Jin Ho; Han, Changpyo; Lee, Dong Wook; Sim, Gyu Hun; Moon, Hye Yun; Kim, Jae-Young; Song, Ji-Yoon; Kang, Hyun Ah

    2018-04-01

    Pyruvate decarboxylase (Pdc) is a cytosolic enzyme located at the branch point between fermentative and respiratory sugar catabolism. Here, we identified and functionally characterized KmPDC1 and KmPDC5 encoding two homologs of Pdc in the thermotolerant yeast Kluyveromyces marxianus KCTC 17555. Despite the conservation of important Pdc domains, a few amino acid sequences essential for enzymatic activity are not conserved in KmPdc5p. Deletion of KmPDC1 alone eliminated most of Pdc activity, but the growth of the Kmpdc1Δ strain on glucose was comparable to that of the wild type (WT) strain under aerobic conditions. In contrast to the WT, Kmpdc1Δ could not grow on glucose under oxygen-limited conditions. The KmPDC5 deletion did not generate any apparent change in Pdc activity or growth patterns under several tested conditions. Whereas the expression of KmPDC1 was enhanced by glucose, the basic expression levels of KmPDC5 were very low, without a detectable difference between glucose and nonfermentable carbon sources. Moreover, KmPDC5 overexpression was unable to complement the growth defect of Kmpdc1Δ in the presence of antimycin A, and the purified recombinant KmPdc5p was inactive in Pdc activity assay, supporting the notion that KmPdc5p may lack Pdc enzymatic activity. Notably, compared to the WT, Kmpdc1Δ single and Kmpdc1Δpdc5Δ double mutants produced significantly less glycerol, acetate, and ethanol while accumulating pyruvate. Altogether, our data indicate that a single deletion of KmPDC1 is sufficient in Crabtree-negative K. marxianus strains to generate a starting host strain for engineering of production of high-value biomaterials derived from pyruvate without byproduct formation.

  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. 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...

  9. 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

  10. 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.

  11. 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

  12. 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

  13. 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.

  14. Occurrence of Co-Infection of Helicobacter pullorum and Campylobacter spp. in Broiler and Village (Indigenous Chickens

    Directory of Open Access Journals (Sweden)

    Soe Soe Wai, A. A. Saleha*, Z. Zunita, L. Hassan and A. Jalila

    2012-10-01

    Full Text Available The reports on prevalence of Helicobacter pullorum in broiler chickens are rather limited and lacking in village chickens. This study aimed to determine the occurrence of H. pullorum in broiler and village chickens in Selangor, Malaysia and to report the detection of co-infection of H. pullorum and Campylobacter spp. in these chickens. Village (indigenous chickens were sampled in five markets and broiler chickens from six farms in different localities. Cecal contents were aseptically obtained from the chickens and subjected to three cultural methods. The isolates were identified by biochemical tests and confirmed using a species-specific PCR assay. Helicobacter pullorum were isolated from 25% village chickens and 24.6% broiler chickens, with an overall occurrence of 24.7%. Eleven (50% of these positive chickens (nine in broiler and two in village chickens showed co-infection with Campylobacter spp.

  15. Participation of some campylobacter species in the etiology of enterocolitis

    Directory of Open Access Journals (Sweden)

    Otašević Marica M.

    2004-01-01

    Full Text Available Background. In recent decades, medical community has increasingly been calling attention to the importance of Campylobacter as an disease-causing agent in humans. Nowdays, Campylobacter jejuni (C. jejuni is known as the most frequent bacterial cause of diarrhea worldwide. Epidemiological differences of the infections caused by Campylobacter, present in the developed and the developing countries, are attributed to the differences of the types of virulence. Due to the specificity, and the demanding features of Campylobacter, as well as poorly equipped microbiological laboratories, campylobacteriosis is insufficiently studied in our country. This investigation aimed to determine the participation of some Campylobacter species in the etiology of diarrheal diseases in our population. Methods. The four-years continuous monitoring of Campylobacter presence was performed in the faeces of 12 605 patients with enterocolitis. The control group included 5 774 examinees of healthy children and youth. Faeces samples were cultivated on Skirrow's selective medium, and further incubated according to effective methodology for Campylobacter. Identification of strains was based on morphological, cultural and physiologic features of strains (oxidase test, catalase test, susceptibility to nalidixic acid, and hypurate hydrolysis. As a statistical method, for data processing, c2 test and Fisher’s exact test were used. Results. Campylobacter was proven in 3.86% of enterocolitis patients, and in 0.71% of healthy population. Out of 518 Campylobacter isolates, 86.48% belonged to enterocolitis outpatients, and 13,51% to inpatients. Predominant symptoms of the disease were diarrhea (81.83%, increased temperature (34.71%, vomiting (19.77%, and stomach pain (15.17%. The diseased were predominantly infants in the first year of life. Out of 300 Campylobacter isolates, 75% were identified as Campylobacer jejuni, 23% as Campylobacter coli (C. coli, and 2% as Campylobacter lari

  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. High Prevalence of Campylobacter ureolyticus in Stool Specimens of Children with Diarrhea in Japan.

    Science.gov (United States)

    Hatanaka, Noritoshi; Shimizu, Akinori; Somroop, Srinuan; Li, Yiming; Asakura, Masahiro; Nagita, Akira; Prasad Awasthi, Sharda; Hinenoya, Atsushi; Yamasaki, Shinji

    2017-07-24

    Campylobacter ureolyticus has been considered as a potentially pathogenic bacterium. In this study, a total of 586 stool samples were collected from 0-12-year-old children with diarrhea between November 2013 and April 2015 and examined with microbiological tests in the hospital for the diagnosis of common enteric pathogens including C. jejuni and C. coli. Then in our laboratory, these samples were analyzed by 16S rRNA sequence-based Campylobacter genus-specific PCR (C16S PCR); 283 (48.3%) samples showed positive results with this PCR assay. Furthermore, C. ureolyticus was screened in these 283 samples by PCR assay, which can detect this species specifically. Surprisingly, C. ureolyticus was detected in 147 of the 283 C16S PCR-positive diarrheal stool samples (51.9%), which is much higher than the prevalence of C. jejuni and C. coli (15.5%), and 96 samples out of 147 were negative for any of the other enteric pathogens tested in the hospital; namely, C. ureolyticus was detected as a single pathogen in 96 samples. This finding suggests that C. ureolyticus may be a pathogen associated with diarrhea in children in Japan. To the best of our knowledge, this is the first report in which C. ureolyticus was detected among Japanese children with diarrhea.

  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. 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.

  20. Host-Pathogen Interactions in Campylobacter Infections: the Host Perspective

    Science.gov (United States)

    Janssen, Riny; Krogfelt, Karen A.; Cawthraw, Shaun A.; van Pelt, Wilfrid; Wagenaar, Jaap A.; Owen, Robert J.

    2008-01-01

    Campylobacter is a major cause of acute bacterial diarrhea in humans worldwide. This study was aimed at summarizing the current understanding of host mechanisms involved in the defense against Campylobacter by evaluating data available from three sources: (i) epidemiological observations, (ii) observations of patients, and (iii) experimental observations including observations of animal models and human volunteer studies. Analysis of available data clearly indicates that an effective immune system is crucial for the host defense against Campylobacter infection. Innate, cell-mediated, and humoral immune responses are induced during Campylobacter infection, but the relative importance of these mechanisms in conferring protective immunity against reinfection is unclear. Frequent exposure to Campylobacter does lead to the induction of short-term protection against disease but most probably not against colonization. Recent progress in the development of more suitable animal models for studying Campylobacter infection has opened up possibilities to study the importance of innate and adaptive immunity during infection and in protection against reinfection. In addition, advances in genomics and proteomics technologies will enable more detailed molecular studies. Such studies combined with better integration of host and pathogen research driven by epidemiological findings may truly advance our understanding of Campylobacter infection in humans. PMID:18625685

  1. Campylobacter species in animal, food, and environmental sources, and relevant testing programs in Canada.

    Science.gov (United States)

    Huang, Hongsheng; Brooks, Brian W; Lowman, Ruff; Carrillo, Catherine D

    2015-10-01

    Campylobacter species, particularly thermophilic campylobacters, have emerged as a leading cause of human foodborne gastroenteritis worldwide, with Campylobacter jejuni, Campylobacter coli, and Campylobacter lari responsible for the majority of human infections. Although most cases of campylobacteriosis are self-limiting, campylobacteriosis represents a significant public health burden. Human illness caused by infection with campylobacters has been reported across Canada since the early 1970s. Many studies have shown that dietary sources, including food, particularly raw poultry and other meat products, raw milk, and contaminated water, have contributed to outbreaks of campylobacteriosis in Canada. Campylobacter spp. have also been detected in a wide range of animal and environmental sources, including water, in Canada. The purpose of this article is to review (i) the prevalence of Campylobacter spp. in animals, food, and the environment, and (ii) the relevant testing programs in Canada with a focus on the potential links between campylobacters and human health in Canada.

  2. Preliminary physiological characteristics of thermotolerant Saccharomyces cerevisiae clinical isolates identified by molecular biology techniques.

    Science.gov (United States)

    Siedlarz, P; Sroka, M; Dyląg, M; Nawrot, U; Gonchar, M; Kus-Liśkiewicz, M

    2016-03-01

    The aim of the study was a molecular identification and physiological characteristic of the five Saccharomyces cerevisiae strains isolated from patients. The tested isolates were compared with control strains (which are of laboratory or commercial origin). The relation of the isolates to baker's yeast S. cerevisiae was studied using species-specific primers in PCR analysis of the ITS-26S region of DNA. Five isolates were genetically identified as the yeast belonging to the genus S. cerevisiae. The effects of temperature and carbon sources on the growth of the yeast strains were analysed. A quantitative characterization of growth kinetics approve that some tested isolates are thermotolerant and are able to grow at range 37-39°C. Among them, one representative is characterized by the highest specific growth rate (0·637 h(-1) ). In conclusions, some strains are defined as potential candidates to use in the biotechnology due to a higher growth rate at elevated temperatures. Screening for further evaluation of biotechnological significance of the tested isolates will be done (e.g. ethanol and trehalose production at higher temperatures). The physiological characterization and confirmation of species identification by molecular methods for yeasts important in the context of biotechnology industry were demonstrated. Thermotolerant microbial strains are required in various industrial applications, for improving productivity and for decreasing the risk of undesirable contaminations when higher temperatures are used. It is important to search for such strains in extreme environments or exotic niches. In this paper, new thermotolerant strains were identified belonging to the Saccharomyces cerevisiae, but differed from typical bakers' yeast, essentially by their growth rate at higher temperature. The described yeast strains are promising for using in biotechnological industry, especially, for production of ethanol and other products at higher temperatures. © 2015 The

  3. High-temperature ethanol production using thermotolerant yeast newly isolated from Greater Mekong Subregion.

    Science.gov (United States)

    Techaparin, Atiya; Thanonkeo, Pornthap; Klanrit, Preekamol

    The application of high-potential thermotolerant yeasts is a key factor for successful ethanol production at high temperatures. Two hundred and thirty-four yeast isolates from Greater Mekong Subregion (GMS) countries, i.e., Thailand, The Lao People's Democratic Republic (Lao PDR) and Vietnam were obtained. Five thermotolerant yeasts, designated Saccharomyces cerevisiae KKU-VN8, KKU-VN20, and KKU-VN27, Pichia kudriavzevii KKU-TH33 and P. kudriavzevii KKU-TH43, demonstrated high temperature and ethanol tolerance levels up to 45°C and 13% (v/v), respectively. All five strains produced higher ethanol concentrations and exhibited greater productivities and yields than the industrial strain S. cerevisiae TISTR5606 during high-temperature fermentation at 40°C and 43°C. S. cerevisiae KKU-VN8 demonstrated the best performance for ethanol production from glucose at 37°C with an ethanol concentration of 72.69g/L, a productivity of 1.59g/L/h and a theoretical ethanol yield of 86.27%. The optimal conditions for ethanol production of S. cerevisiae KKU-VN8 from sweet sorghum juice (SSJ) at 40°C were achieved using the Box-Behnken experimental design (BBD). The maximal ethanol concentration obtained during fermentation was 89.32g/L, with a productivity of 2.48g/L/h and a theoretical ethanol yield of 96.32%. Thus, the newly isolated thermotolerant S. cerevisiae KKU-VN8 exhibits a great potential for commercial-scale ethanol production in the future. Copyright © 2017 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.

  4. Bioprospecting thermotolerant ethanologenic yeasts for simultaneous saccharification and fermentation from diverse environments.

    Science.gov (United States)

    Choudhary, Jairam; Singh, Surender; Nain, Lata

    2017-03-01

    Lignocellulosic biomass, a promising renewable energy source, can be used for the production of second generation bioethanol. Simultaneous saccharification and fermentation (SSF), the process which alleviates the problem of separate hydrolysis and fermentation (SHF), requires thermotolerant ethanologenic yeast for bioethanol production. Therefore, ten yeast strains isolated from diverse sources, belonging to various genera like Saccharomyces, Candida, Pichia and Wickerhamomyces were evaluated for their thermotolerance, sugar utilization pattern, inhibitor tolerance and ethanol production potential with glucose, xylose and alkali pretreated paddy straw. All the tested strains were found to be thermotolerant, capable of significant growth at 40°C. Candida tropicalis Y6 was capable of utilizing a wide range of sugars as compared with other yeast isolates. Strains of Candida showed better inhibitor tolerance as compared to Saccharomyces and Pichia strains and exhibited only 5.1-18.8% and 4.7-7.9% reduction in growth with furfural and 5-hydroxymethyl furfural, respectively. Saccharomyces cerevisiae JRC6, isolated from distillery waste, produced ethanol with 88.3% and 89.1% theoretical efficiency at 40°C and 42°C, respectively, from glucose. This strain also produced significantly higher amount of ethanol (3.8 g/L) with better fermentation efficiency (87.9%) from alkali pretreated paddy straw at 40°C, as compared with the other yeast strains. Therefore, S. cerevisiae JRC6, based on its ability to ferment sugars at a higher temperature, can be a promising candidate for production of ethanol from lignocellulosic biomass via SSF process. Copyright © 2016 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  5. High-temperature ethanol production using thermotolerant yeast newly isolated from Greater Mekong Subregion

    Directory of Open Access Journals (Sweden)

    Atiya Techaparin

    Full Text Available Abstract The application of high-potential thermotolerant yeasts is a key factor for successful ethanol production at high temperatures. Two hundred and thirty-four yeast isolates from Greater Mekong Subregion (GMS countries, i.e., Thailand, The Lao People's Democratic Republic (Lao PDR and Vietnam were obtained. Five thermotolerant yeasts, designated Saccharomyces cerevisiae KKU-VN8, KKU-VN20, and KKU-VN27, Pichia kudriavzevii KKU-TH33 and P. kudriavzevii KKU-TH43, demonstrated high temperature and ethanol tolerance levels up to 45 °C and 13% (v/v, respectively. All five strains produced higher ethanol concentrations and exhibited greater productivities and yields than the industrial strain S. cerevisiae TISTR5606 during high-temperature fermentation at 40 °C and 43 °C. S. cerevisiae KKU-VN8 demonstrated the best performance for ethanol production from glucose at 37 °C with an ethanol concentration of 72.69 g/L, a productivity of 1.59 g/L/h and a theoretical ethanol yield of 86.27%. The optimal conditions for ethanol production of S. cerevisiae KKU-VN8 from sweet sorghum juice (SSJ at 40 °C were achieved using the Box-Behnken experimental design (BBD. The maximal ethanol concentration obtained during fermentation was 89.32 g/L, with a productivity of 2.48 g/L/h and a theoretical ethanol yield of 96.32%. Thus, the newly isolated thermotolerant S. cerevisiae KKU-VN8 exhibits a great potential for commercial-scale ethanol production in the future.

  6. Production of pullulan by a thermotolerant Aureobasidium pullulans strain in non-stirred fed batch fermentation process

    OpenAIRE

    Singh, Ranjan; Gaur, Rajeeva; Tiwari, Soni; Gaur, Manogya Kumar

    2012-01-01

    Total 95 isolates of Aureobasidium pullulans were isolated from different flowers and leaves samples, out of which 11 thermotolerant strains produced pullulan. One thermotolerant non-melanin pullulan producing strain, designated as RG-5, produced highest pullulan (37.1±1.0 g/l) at 42ºC, pH 5.5 in 48h of incubation with 3% sucrose and 0.5% ammonium sulphate in a non-stirred fed batch fermentor of 6 liters capacity. The two liters of initial volume of fermentation medium was further fed with th...

  7. Thermotolerant yeasts selected by adaptive evolution express heat stress response at 30ºC

    DEFF Research Database (Denmark)

    Caspeta, Luis; Chen, Yun; Nielsen, Jens

    2016-01-01

    to grow at increased temperature, activated a constitutive heat stress response when grown at the optimal ancestral temperature, and that this is associated with a reduced growth rate. This preventive response was perfected by additional transcriptional changes activated when the cultivation temperature...... temperatures, but this also causes a trade-off in the growth rate at the optimal ancestral temperature.......Exposure to long-term environmental changes across >100s of generations results in adapted phenotypes, but little is known about how metabolic and transcriptional responses are optimized in these processes. Here, we show that thermotolerant yeast strains selected by adaptive laboratory evolution...

  8. Sea urchin development in a global change hotspot, potential for southerly migration of thermotolerant propagules

    Science.gov (United States)

    Byrne, M.; Selvakumaraswamy, P.; Ho, M. A.; Woolsey, E.; Nguyen, H. D.

    2011-03-01

    The distribution of the sea urchin Heliocidaris erythrogramma coincides with the southeast Australia global change hot spot where marine ecosystems are warming significantly due to changes in ocean circulation. To address questions on future vulnerabilities, the thermotolerance of the planktonic life phase of H. erythrogramma was investigated in the climate and regionally relevant setting of projected near-future (2100) ocean warming. Experimental treatments ranged from 18 to 26 °C, with 26 °C representing +3-4 °C above recent ambient sea-surface temperatures. Developmental success across all stages (gastrula, 24 h; larva, 72 h; juvenile, 120 h) decreased with increasing temperature. Development was tolerant to a +1-2 °C increase above ambient, but significant deleterious effects were evident at +3-4 °C. However, larvae that developed through the early bottleneck of normal development at 26 °C metamorphosed successfully. The inverse relationship between temperature and planktonic larval duration (PLD) was seen in a 25% decrease in the PLD of H. erythrogramma at 24 and 26 oC. Ocean warming may be advantageous to a subset of larvae through early settlement and reduction of the vulnerable planktonic period. This positive effect of temperature may help buffer the negative effects of ocean warming. In parallel studies with progeny derived from northern (Coffs Harbour) and southern (Sydney) H. erythrogramma, northern embryos had significantly higher thermotolerance. This provides the possibility that H. erythrogramma populations might keep up with a warming world through poleward migration of thermotolerant propagules, facilitated by the strong southward flow of the East Australian Current. It is uncertain whether H. erythrogramma populations at the northern range of this species, with no source of immigrants, will have the capacity to persist in a warm ocean. Due to its extensive latitudinal distribution, its potential developmental thermotolerance and

  9. Prevalence of Campylobacter concisus in diarrhoea of immunocompromised patients

    DEFF Research Database (Denmark)

    Aabenhus, Rune Munck; Permin, Henrik; On, Stephen L W

    2002-01-01

    The importance of Campylobacter species other than C. jejuni/coli in diarrhoeal disease is largely unknown. We wished to determine the prevalence and clinical presentation of C. concisus infection in patients with enteric disease in a tertiary hospital. Stool specimens were routinely tested...... for the presence of Campylobacter species, by use of the filter isolation method. The medical records of the C. concisus-positive patients were reviewed. Of 224 Campylobacter isolates obtained, 110 were identified as C. concisus. Concomitant infection occurred in only 27% of cases. By means of protein profiling we...

  10. Prevalence of Campylobacter concisus in diarrhoea of immunocompromised patients

    DEFF Research Database (Denmark)

    Aabenhus, Rune; Permin, Henrik; On, Stephen L.W.

    2002-01-01

    The importance of Campylobacter species other than C. jejuni/coli in diarrhoeal disease is largely unknown. We wished to determine the prevalence and clinical presentation of C. concisus infection in patients with enteric disease in a tertiary hospital. Stool specimens were routinely tested...... for the presence of Campylobacter species, by use of the filter isolation method. The medical records of the C. concisus-positive patients were reviewed. Of 224 Campylobacter isolates obtained, 110 were identified as C. concisus. Concomitant infection occurred in only 27% of cases. 13 means of protein profiling we...

  11. Current methods for molecular typing of Campylobacter species.

    Science.gov (United States)

    Taboada, Eduardo N; Clark, Clifford G; Sproston, Emma L; Carrillo, Catherine D

    2013-10-01

    Campylobacter remains one of the most common bacterial causes of gastroenteritis worldwide. Tracking sources of this organism is challenging due to the large numbers of human cases, and the prevalence of this organism throughout the environment due to growth in a wide range of animal species. Many molecular subtyping methods have been developed to characterize Campylobacter species, but only a few are commonly used in molecular epidemiology studies. This review examines the applicability of these methods, as well as the role that emerging whole genome sequencing technologies will play in tracking sources of Campylobacter spp. infection. © 2013.

  12. Campylobacter spp. as a foodborne pathogen: a review

    Directory of Open Access Journals (Sweden)

    Joana eSilva

    2011-09-01

    Full Text Available Campylobacter is well recognized as the leading cause of bacterial foodborne diarrheal disease worldwide, causing mild to severe symptoms including serious infections of the extremities and permanent neurological symptoms. The organism is a cytochrome oxidase positive, microaerophilic, curved Gram-negative rod exhibiting corkscrew motility and is carried in the intestine of many wild and domestic animals, particularly avian species including poultry, where the intestine is colonized resulting in healthy animals as carriers. This review aims to elucidate and discuss the i genus Campylobacter, growth and survival characteristics; ii detection, isolation and confirmation of Campylobacter; iii campylobacteriosis and presence of virulence factors and iv colonization of poultry and control strategies.

  13. 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.

  14. Soluble and cell-associated haemagglutinins of Helicobacter (Campylobacter) pylori.

    Science.gov (United States)

    Robinson, J; Goodwin, C S; Cooper, M; Burke, V; Mee, B J

    1990-12-01

    Some plate-grown strains of Helicobacter (Campylobacter) pylori that were harvested into phosphate-buffered saline and left for 1 h released soluble haemagglutinins. These caused high-titre agglutination of human and guinea-pig erythrocytes, whereas chicken, sheep and bovine erythrocytes were agglutinated at various titres. Six of 10 strains which had been subcultured repeatedly did not possess soluble haemagglutinins. Slide agglutination of bacterial suspensions demarcated the strains into two groups; Group 1 gave strong agglutination with most types of erythrocyte, Group 2 did not. By microtitration assay, all Group-1 strains but only two Group-2 strains produced a soluble haemagglutinin. Cell-associated haemagglutinins were found by microtitration assay in all strains of H. pylori, but higher titres were found within Group-1 strains. The supernates of broth-grown, shaken cultures also showed the presence of soluble haemagglutinins, with higher titres for recently isolated strains. Pre-treatment of human erythrocytes with neuraminidase from Arthrobacter ureafaciens and Clostridium perfringens abolished haemagglutination by the soluble, but not by the cell-associated haemagglutinin. The soluble haemagglutinin was inhibited by sialoproteins containing predominantly the N-acetylneuraminyl (2-3) galactopyranosyl [NeuAc(2-3)Gal] structure, fetuin, glycophorin and bovine N-acetylneuraminyl-lactose (NeuAc-Lac). Transferrin and human NeuAc-Lac, which contain predominantly the N-acetylneuraminyl (2-6) galactopyranosyl [NeuAc(2-6)Gal] structure were not inhibitory. However, bovine submaxillary mucin (BSM) was strongly inhibitory; it contains several structures with sialic acid linked 2-6 to oligosaccharides. These results suggest that the soluble haemagglutinin recognises a NeuAc(2-3)Gal structure, but has high affinity for another, as yet undetermined, sialic acid-containing structure.

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

    Science.gov (United States)

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

  16. Molecular Typing ofCampylobacter jejuniandCampylobacter coliIsolated from Various Retail Meats by MLST and PFGE.

    Science.gov (United States)

    Noormohamed, Aneesa; Fakhr, Mohamed K

    2014-01-08

    Campylobacter species are one of the leading causes of foodborne disease in the United States. Campylobacter jejuni and Campylobacter coli are the two main species of concern to human health and cause approximately 95% of human infections. Molecular typing methods, such as pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) are often used to source track foodborne bacterial pathogens. The aim of the present study was to compare PFGE and MLST in typing strains of C. jejuni and C. coli that were isolated from different Oklahoma retail meat sources. A total of 47 Campylobacter isolates (28 C. jejuni and 19 C. coli ) isolated from various retail meat samples (beef, beef livers, pork, chicken, turkey, chicken livers, and chicken gizzards) were subjected to pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). PFGE was able to group the 47 Campylobacter isolates into two major clusters (one for C. jejuni and one for C. coli ) but failed to differentiate the isolates according to their source. MLST revealed 21 different sequence types (STs) that belonged to eight different clonal complexes. Twelve of the screened Campylobacter isolates (8 C. jejuni and 4 C. coli ) did not show any defined STs. All the defined STs of C. coli isolates belonged to ST-828 complex. The majority of C. jejuni isolates belonged to ST-353, ST-607, ST-52, ST-61, and ST-21 complexes. It is worthy to mention that, while the majority of Campylobacter isolates in this study showed STs that are commonly associated with human infections along with other sources, most of the STs from chicken livers were solely reported in human cases. In conclusion, retail meat Campylobacter isolates tested in this study particularly those from chicken livers showed relatedness to STs commonly associated with humans. Molecular typing, particularly MLST, proved to be a helpful tool in suggesting this relatedness to Campylobacter human isolates.

  17. 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.

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

    Directory of Open Access Journals (Sweden)

    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.

  19. Same-Day Subtyping of Campylobacter jejuni and C. coli Isolates by Use of Multiplex Ligation-Dependent Probe Amplification–Binary Typing

    Science.gov (United States)

    Vandenberg, Olivier; Robson, Beth; Gilpin, Brent J.; Brandt, Stephanie M.; Scholes, Paula; Martiny, Delphine; Carter, Philip E.; van Vught, Paul; Schouten, Jan; On, Stephen L. W.

    2014-01-01

    Campylobacteriosis is the most commonly reported form of human bacterial gastroenteritis in the world. Sound identification of infectious sources requires subtyping, but the most widely used methods have turnaround times measured in days and require specialist equipment and skills. A multiplex ligation-dependent probe amplification-binary typing (MBiT) assay was developed for subtyping Campylobacter jejuni and Campylobacter coli. It was tested on 245 isolates, including recent isolates from Belgium and New Zealand, and compared to multilocus sequence typing (MLST). When used in an outbreak setting, MBiT identified the predominant genotype and possible additional cases days before pulsed-field gel electrophoresis (PFGE) results were available. MBiT was more discriminatory than MLST and, being a single assay with results produced within 6 h, was more rapid and cost-effective than both MLST and PFGE. In addition, MBiT requires only basic molecular biology equipment and skills. PMID:24989612

  20. Estimating sensitivity and specificity of a PCR for boot socks to detect Campylobacter in broiler primary production using Bayesian latent class analysis

    DEFF Research Database (Denmark)

    Matt, Monika; Nordentoft, Steen; Ian, Kopacka

    2016-01-01

    The present study compares three different assays for sample collection and detection of Campylobacter spp. in broiler flocks, based on (i) the collection of faecal samples from intestinal organs (caecum), (ii) individual faecal droppings collected from the bedding and (iii) faecal material...... collected by socks placed on the outside of a pair of boots (boot socks) and used for walking around in the flock. The two first methods are examined for Campylobacter using a culture method (ISO-10272-2:2006), while the boot socks are tested using PCR. The PCR-assay is a genus specific multiplex PCR...... samples were collected at slaughter.The results were evaluated in the absence of a gold standard using a Bayesian latent class model. Austrian results showed higher sensitivity for PCR detection in sock samples (0.98; Bayesian credible interval (BCI) [0.93-1]) than for culture of faecal droppings (0...

  1. Thermo-tolerant phosphate-solubilizing microbes for multi-functional biofertilizer preparation.

    Science.gov (United States)

    Chang, Cheng-Hsiung; Yang, Shang-Shyng

    2009-02-01

    In order to prepare the multi-functional biofertilizer, thermo-tolerant phosphate-solubilizing microbes including bacteria, actinomycetes, and fungi were isolated from different compost plants and biofertilizers. Except Streptomycesthermophilus J57 which lacked pectinase, all isolates possessed amylase, CMCase, chitinase, pectinase, protease, lipase, and nitrogenase activities. All isolates could solubilize calcium phosphate and Israel rock phosphate; various isolates could solubilize aluminum phosphate, iron phosphate, and hydroxyapatite. During composting, biofertilizers inoculated with the tested microbes had a significantly higher temperature, ash content, pH, total nitrogen, soluble phosphorus content, and germination rate than non-inoculated biofertilizer; total organic carbon and carbon-to-nitrogen ratio showed the opposite pattern. Adding these microbes can shorten the period of maturity, improve the quality, increase the soluble phosphorus content, and enhance the populations of phosphate-solubilizing and proteolytic microbes in biofertilizers. Therefore, inoculating thermo-tolerant phosphate-solubilizing microbes into agricultural and animal wastes represents a practical strategy for preparing multi-functional biofertilizer.

  2. The growth profile, thermotolerance and biofilm formation of Enterobacter sakazakii grown in infant formula milk.

    Science.gov (United States)

    Iversen, C; Lane, M; Forsythe, S J

    2004-01-01

    To study the growth, thermotolerance and biofilm formation of the emergent pathogen Enterobacter sakazakii in infant formula milk (IFM). The temperature range, death kinetics and biofilm formation of E. sakazakii were determined using impedance microbiology and conventional methods. In IFM the organism grew as low as 6 degrees C and optimally at 37-43 degrees C. In faecal coliform tests, 23% of strains (n = 70) produced gas from lauryl sulphate broth (LSB) at 44 degrees C after 48 h incubation. Three strains failed to grow in LSB at any of the temperatures. The D-value of cells suspended in IFM was determined between 54 and 62 degrees C. The resultant z-value was 5.7 degrees C. The organism was able to adhere and grow on latex, polycarbonate, silicon and to a lesser extent stainless steel. Enterobacter sakazakii was able to grow at refrigeration temperatures and on infant-feeding equipment. The thermotolerance of the organism was similar to other Enterobacteriaceae and should be killed during standard pasteurization treatment. Enterobacter sakazakii has been associated with infant meningitis through consumption of contaminated IFM. Enterobacter sakazakii is able to grow in IFM during storage at refrigeration temperatures and attach to infant-feeding equipment, which may become reservoirs of infection.

  3. Changes in Salicylic Acid and Antioxidants during Induced Thermotolerance in Mustard Seedlings

    Science.gov (United States)

    Dat, James F.; Foyer, Christine H.; Scott, Ian M.

    1998-01-01

    Heat-acclimation or salicylic acid (SA) treatments were previously shown to induce thermotolerance in mustard (Sinapis alba L.) seedlings from 1.5 to 4 h after treatment. In the present study we investigated changes in endogenous SA and antioxidants in relation to induced thermotolerance. Thirty minutes into a 1-h heat-acclimation treatment glucosylated SA had increased 5.5-fold and then declined during the next 6 h. Increases in free SA were smaller (2-fold) but significant. Changes in antioxidants showed the following similarities after either heat-acclimation or SA treatment. The reduced-to-oxidized ascorbate ratio was 5-fold lower than the controls 1 h after treatment but recovered by 2 h. The glutathione pool became slightly more oxidized from 2 h after treatment. Glutathione reductase activity was more than 50% higher during the first 2 h. Activities of dehydroascorbate reductase and monodehydroascorbate reductase decreased by at least 25% during the first 2 h but were 20% to 60% higher than the control levels after 3 to 6 h. One hour after heat acclimation ascorbate peroxidase activity was increased by 30%. Young leaves appeared to be better protected by antioxidant enzymes following heat acclimation than the cotyledons or stem. Changes in endogenous SA and antioxidants may be involved in heat acclimation. PMID:9847121

  4. Salinity modulates thermotolerance, energy metabolism and stress response in amphipodsGammarus lacustris.

    Science.gov (United States)

    Vereshchagina, Kseniya P; Lubyaga, Yulia A; Shatilina, Zhanna; Bedulina, Daria; Gurkov, Anton; Axenov-Gribanov, Denis V; Baduev, Boris; Kondrateva, Elizaveta S; Gubanov, Mikhail; Zadereev, Egor; Sokolova, Inna; Timofeyev, Maxim

    2016-01-01

    Temperature and salinity are important abiotic factors for aquatic invertebrates. We investigated the influence of different salinity regimes on thermotolerance, energy metabolism and cellular stress defense mechanisms in amphipods Gammarus lacustris Sars from two populations. We exposed amphipods to different thermal scenarios and determined their survival as well as activity of major antioxidant enzymes (peroxidase, catalase, glutathione S-transferase) and parameters of energy metabolism (content of glucose, glycogen, ATP, ADP, AMP and lactate). Amphipods from a freshwater population were more sensitive to the thermal challenge, showing higher mortality during acute and gradual temperature change compared to their counterparts from a saline lake. A more thermotolerant population from a saline lake had high activity of antioxidant enzymes. The energy limitations of the freshwater population (indicated by low baseline glucose levels, downward shift of the critical temperature of aerobic metabolism and inability to maintain steady-state ATP levels during warming) was observed, possibly reflecting a trade-off between the energy demands for osmoregulation under the hypo-osmotic condition of a freshwater environment and protection against temperature stress.

  5. Salinity modulates thermotolerance, energy metabolism and stress response in amphipods Gammarus lacustris

    Directory of Open Access Journals (Sweden)

    Kseniya P. Vereshchagina

    2016-11-01

    Full Text Available Temperature and salinity are important abiotic factors for aquatic invertebrates. We investigated the influence of different salinity regimes on thermotolerance, energy metabolism and cellular stress defense mechanisms in amphipods Gammarus lacustris Sars from two populations. We exposed amphipods to different thermal scenarios and determined their survival as well as activity of major antioxidant enzymes (peroxidase, catalase, glutathione S-transferase and parameters of energy metabolism (content of glucose, glycogen, ATP, ADP, AMP and lactate. Amphipods from a freshwater population were more sensitive to the thermal challenge, showing higher mortality during acute and gradual temperature change compared to their counterparts from a saline lake. A more thermotolerant population from a saline lake had high activity of antioxidant enzymes. The energy limitations of the freshwater population (indicated by low baseline glucose levels, downward shift of the critical temperature of aerobic metabolism and inability to maintain steady-state ATP levels during warming was observed, possibly reflecting a trade-off between the energy demands for osmoregulation under the hypo-osmotic condition of a freshwater environment and protection against temperature stress.

  6. Salinity modulates thermotolerance, energy metabolism and stress response in amphipods Gammarus lacustris

    Science.gov (United States)

    Vereshchagina, Kseniya P.; Lubyaga, Yulia A.; Shatilina, Zhanna; Bedulina, Daria; Gurkov, Anton; Axenov-Gribanov, Denis V.; Baduev, Boris; Kondrateva, Elizaveta S.; Gubanov, Mikhail; Zadereev, Egor; Sokolova, Inna

    2016-01-01

    Temperature and salinity are important abiotic factors for aquatic invertebrates. We investigated the influence of different salinity regimes on thermotolerance, energy metabolism and cellular stress defense mechanisms in amphipods Gammarus lacustris Sars from two populations. We exposed amphipods to different thermal scenarios and determined their survival as well as activity of major antioxidant enzymes (peroxidase, catalase, glutathione S-transferase) and parameters of energy metabolism (content of glucose, glycogen, ATP, ADP, AMP and lactate). Amphipods from a freshwater population were more sensitive to the thermal challenge, showing higher mortality during acute and gradual temperature change compared to their counterparts from a saline lake. A more thermotolerant population from a saline lake had high activity of antioxidant enzymes. The energy limitations of the freshwater population (indicated by low baseline glucose levels, downward shift of the critical temperature of aerobic metabolism and inability to maintain steady-state ATP levels during warming) was observed, possibly reflecting a trade-off between the energy demands for osmoregulation under the hypo-osmotic condition of a freshwater environment and protection against temperature stress. PMID:27896024

  7. Production of Bioethanol from Carrot Pomace Using the Thermotolerant Yeast Kluyveromyces marxianus

    Energy Technology Data Exchange (ETDEWEB)

    Chi-Yang Yu; Bo-Hong Jiang; Kow-Jen Duan [Tatung University, Tapei, Taiwan (China). Department of Bioengineering

    2013-03-15

    Carrot pomace, a major agricultural waste from the juice industry, was used as a feedstock for bioethanol production by fermentation with the thermotolerant yeast Kluyveromyces marxianus. Treatment of the carrot pomace with Accellerase(TM) 1000 and pectinase at 50 °C for 84 h, resulted in conversion of 42% of its mass to fermentable sugars, mainly glucose, fructose, and sucrose. Simultaneous saccharification and fermentation (SSF) at 42 °C was performed on 10% (w/v) carrot pomace; the concentration of ethanol reached 18 g/L and the yield of ethanol from carrot pomace was 0.18 g/g. The highest ethanol concentration of 37 g/L was observed with an additional charge of 10% supplemented to the original 10% of carrot pomace after 12 h; the corresponding yield was 0.185 g/g. Our results clearly demonstrated the potential of combining a SSF process with thermotolerant yeast for the production of bioethanol using carrot pomace as a feedstock.

  8. Campylobacter fetus infections in humans : exposure and disease

    NARCIS (Netherlands)

    Wagenaar, Jaap A; van Bergen, Marcel A P; Blaser, Martin J; Tauxe, Robert V; Newell, Diane G; van Putten, Jos P M

    Campylobacter fetus can cause intestinal illness and, occasionally, severe systemic infections. Infections mainly affect persons at higher risk, including elderly and immunocompromised individuals and those with occupational exposure to infected animals. Outbreaks are infrequent but have provided

  9. Spondylodiscitis and an aortic aneurysm due to Campylobacter coli

    Directory of Open Access Journals (Sweden)

    Bournet Béatrice

    2010-02-01

    Full Text Available Abstract Campylobacter coli is a rare cause of bacteremia. We report here the first case of C.coli spondylodiscitis complicated by an aortic aneurysm. Outcome was favourable with surgery and antibiotic therapy.

  10. Salmonella and Campylobacter: Antimicrobial resistance and bacteriophage control in poultry.

    Science.gov (United States)

    Grant, Ar'Quette; Hashem, Fawzy; Parveen, Salina

    2016-02-01

    Salmonella and Campylobacter are major causes of foodborne related illness and are traditionally associated with consuming undercooked poultry and/or consuming products that have been cross contaminated with raw poultry. Many of the isolated Salmonella and Campylobacter that can cause disease have displayed antimicrobial resistance phenotypes. Although poultry producers have reduced on-the-farm overuse of antimicrobials, antimicrobial resistant Salmonella and Campylobacter strains still persist. One method of bio-control, that is producing promising results, is the use of lytic bacteriophages. This review will highlight the current emergence and persistence of antimicrobial resistant Salmonella and Campylobacter recovered from poultry as well as bacteriophage research interventions and limitations. Copyright © 2015 Elsevier Ltd. All rights reserved.

  11. 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.

  12. Biodiversity, ecology, and evolution of Campylobacter in reptiles

    NARCIS (Netherlands)

    Gilbert, M.J.

    2015-01-01

    Species of the Epsilonproteobacteria genera Campylobacter, Arcobacter, and Helicobacter are frequently isolated from endothermic mammals and birds. However, little information was available about the presence of Epsilonproteobacteria in ectothermic reptiles and no comprehensive studies had been

  13. Sensitivity of Campylobacter spp. to irradiation in poultry meat

    International Nuclear Information System (INIS)

    Patterson, M.F.

    1995-01-01

    The sensitivity of Campylobacter jejuni (three strains), Camp. coli (three strains), Camp. fetus (one strain) and Camp. lari (one strain) to irradiation in poultry meat was investigated. There was no significant difference in the counts obtained on Blood or Skirrows agar. Preston agar gave a significantly lower recovery of the pathogens after irradiation so these results were not included in calculations of D10 values. The D10 values ranged from 0.12 to 0.25 kGy and there was a significant difference in the radiation sensitivity between different Campylobacter spp. and within strains of the same species. These values indicate that Campylobacter spp. are more radiation-sensitive than Salmonella and Listeria monocytogenes irradiated under similar conditions. Therefore irradiation treatments suggested to eliminate the latter from poultry carcasses would also be sufficient to remove Campylobacter

  14. Campylobacter spp. as a Foodborne Pathogen: A Review

    Science.gov (United States)

    Silva, Joana; Leite, Daniela; Fernandes, Mariana; Mena, Cristina; Gibbs, Paul Anthony; Teixeira, Paula

    2011-01-01

    Campylobacter is well recognized as the leading cause of bacterial foodborne diarrheal disease worldwide. Symptoms can range from mild to serious infections of the children and the elderly and permanent neurological symptoms. The organism is a cytochrome oxidase positive, microaerophilic, curved Gram-negative rod exhibiting corkscrew motility and is carried in the intestine of many wild and domestic animals, particularly avian species including poultry. Intestinal colonization results in healthy animals as carriers. In contrast with the most recent published reviews that cover specific aspects of Campylobacter/campylobacteriosis, this broad review aims at elucidating and discussing the (i) genus Campylobacter, growth and survival characteristics; (ii) detection, isolation and confirmation of Campylobacter; (iii) campylobacteriosis and presence of virulence factors; and (iv) colonization of poultry and control strategies. PMID:21991264

  15. Occurrence and genotypes of Campylobacter species in broilers during the rearing period.

    Science.gov (United States)

    Zhang, Xiaoyan; Yin, Tiantian; Du, Xueqing; Yang, Wenbin; Huang, Jinlin; Jiao, Xinan

    2017-04-01

    Poultry are the main source of Campylobacter infection worldwide. To obtain information on Campylobacter-infected flocks and create a reference for preventing and controlling Campylobacter at farm level, Campylobacter isolates were recovered from broilers and the environments of nine chicken flocks in two farms during growth. The genetic relationship between the Campylobacter isolates was determined using multilocus sequence typing. Flocks were colonized as early as 3 weeks after introduction to the farm. The highest colonization rate was more than 90% and occurred 4-6 weeks after introduction to the farm. Quantitative data showed that the highest Campylobacter loads appeared at 1-2 weeks after initial colonization. Campylobacter loads in cloacal swabs in four flocks were significantly higher at 5 weeks than at 3 weeks (P Campylobacter jejuni and eight for Campylobacter coli isolates. The STs of the Campylobacter isolates recovered from farm 1 were more diversified than those from farm 2. The STs of environmental samples were highly consistent with those of the cloacal swab samples. The consistency between Campylobacter STs in the environmental and cloacal swab samples suggested that the environment might be one of the main sources of infection. Thus, our study highlights the prevalence and contamination load of Campylobacter in broilers during their rearing period and emphasizes the need for control and prevention measures for Campylobacter infection in broilers, which is also important for human health.

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

    Directory of Open Access Journals (Sweden)

    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%.

  17. "Campylobacter upsaliensis" isolated from blood cultures of pediatric patients.

    OpenAIRE

    Lastovica, A J; Le Roux, E; Penner, J L

    1989-01-01

    Seventeen campylobacters isolated from cultures of blood samples of 16 bacteremic patients were susceptible to cephalothin and were either catalase negative or had only weak catalase activity (CNW strains) and were classified as "Campylobacter upsaliensis" (K. Sandstedt and J. Ursing, XIV Int. Congr. Microbiol., p. B8-17, 1986). All of the patients had predisposing conditions, and 10 patients were less than or equal to 12 months old, indicating that "C. upsaliensis" is an opportunistic pathog...

  18. 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...

  19. 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....

  20. Detection and survival of Campylobacter in chicken eggs.

    Science.gov (United States)

    Sahin, O; Kobalka, P; Zhang, Q

    2003-01-01

    Campylobacter jejuni, a food-borne human pathogen, is widespread in poultry; however, the sources of infection and modes of transmission of this organism on chicken farms are not well understood. The objective of this study was to determine if vertical transmission of C. jejuni occurs via eggs. Using a temperature differential method, it was shown that Campylobacter had limited ability to penetrate the eggshell. When C. jejuni was directly inoculated into the egg yolk and the eggs were stored at 18 degrees C, the organism was able to survive for up to 14 days. However, viability of C. jejuni was dramatically shortened when injected into the albumen or the air sac. When freshly laid eggs from Campylobacter-inoculated specific pathogen-free (SPF) layers were tested, C. jejuni-contamination was detected in three of 65 pooled whole eggs (5-10 eggs in each pool) via culture and PCR. However, the organism was not detected from any of the 800 eggs (80 pools), collected from the same SPF flock, but kept at 18 degrees C for 7 days before testing. Likewise, Campylobacter was not recovered from any of 500 fresh eggs obtained from commercial broiler-breeder flocks that were actively shedding Campylobacter in faeces. Also, none of the 1000 eggs from broiler breeders obtained from a commercial hatchery were positive for Campylobacter. These results suggest that vertical transmission of C. jejuni through the egg is probably a rare event and does not play a major role in the introduction of Campylobacter to chicken flocks. Control of Campylobacter transmission to chicken flocks should focus on sources of infection that are not related to eggs.

  1. Thermotolerance and heat stress responses of Douglas-fir and ponderosa pine seedling populations from contrasting climates

    Science.gov (United States)

    Danielle E. Marias; Frederick C. Meinzer; David R. Woodruff; Katherine A. McCulloh; David Tissue

    2016-01-01

    Temperature and the frequency and intensity of heat waves are predicted to increase throughout the 21st century. Germinant seedlings are expected to be particularly vulnerable to heat stress because they are in the boundary layer close to the soil surface where intense heating occurs in open habitats. We quantified leaf thermotolerance and whole-plant physiological...

  2. Genetically altering the expression of neutral trehalase gene affects conidiospore thermotolerance of the entomopathogenic fungus Metarhizium acridum

    Directory of Open Access Journals (Sweden)

    Peng Guoxiong

    2011-02-01

    Full Text Available Abstract Background The entomopathogenic fungus Metarhizium acridum has been used as an important biocontrol agent instead of insecticides for controlling crop pests throughout the world. However, its virulence varies with environmental factors, especially temperature. Neutral trehalase (Ntl hydrolyzes trehalose, which plays a role in environmental stress response in many organisms, including M. acridum. Demonstration of a relationship between Ntl and thermotolerance or virulence may offer a new strategy for enhancing conidiospore thermotolerance of entomopathogenic fungi through genetic engineering. Results We selected four Ntl over-expression and four Ntl RNA interference (RNAi transformations in which Ntl expression is different. Compared to the wild-type, Ntl mRNA expression was reduced to 35-66% in the RNAi mutants and increased by 2.5-3.5-fold in the over-expression mutants. The RNAi conidiospores exhibited less trehalase activity, accumulated more trehalose, and were much more tolerant of heat stress than the wild-type. The opposite effects were found in conidiospores of over-expression mutants compared to RNAi mutants. Furthermore, virulence was not altered in the two types of mutants compared to the wild type. Conclusions Ntl controlled trehalose accumulation in M. acridum by degrading trehalose, and thus affected conidiospore thermotolerance. These results offer a new strategy for enhancing conidiospore thermotolerance of entomopathogenic fungi without affecting virulence.

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

    Science.gov (United States)

    Chansiripornchai, Niwat; Sasipreeyajan, Jiroj

    2009-06-01

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

  4. 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.

  5. [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.

  6. 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.

  7. 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.

  8. Prevalence and pathogen load of Campylobacter spp., Salmonella enterica and Escherichia coli O157/O145 serogroup in sheep faeces collected at sale yards and in abattoir effluent in Western Australia.

    Science.gov (United States)

    Yang, R; Abraham, S; Gardner, G E; Ryan, U; Jacobson, C

    2017-05-01

    Develop a multiplex quantitative PCR assay to investigate the prevalence and shedding of Escherichia coli O157/O145, Salmonella spp. and Campylobacter spp. in sheep at sale yards and abattoirs. A qPCR for E. coli O157/O145 was developed, validated and multiplexed with an existing qPCR for Campylobacter and Salmonella enterica. The absolute numbers of E. coli O157/O145, Campylobacter and Salmonella in control samples was determined using droplet digital PCR. These were then used as the controls in the multiplex qPCR on a total of 474 sheep faecal samples collected from two saleyards over a 4-month period (April-July 2014) and 96 effluent samples from an abattoir. The mutiplex qPCR was specific with a sensitivity of 5 organisms/μL faecal DNA extract for Campylobacter, S. enterica and E. coli O157/O145. The overall prevalence of Campylobacter, S. enterica and E. coli O157/O145 in faecal samples was 5.7%, 3.6% and 8.4% and in effluent samples was 18.8%, 6.3% and 5.2%, respectively. The pathogen loads of Campylobacter, S. enterica and E. coli O157/O145 in faecal and effluent samples was also determined via mutiplex qPCR. The overall prevalences of Campylobacter, S. enterica and E. coli O157/O145 were generally low (<6%), but point prevalences ranged considerably in healthy sheep (up to 26% for E. coli O157/O145). Further work to determine risk factors for shedding of bacterial organisms in meat sheep in the pre-slaughter period (on-farm, sale yards and lairage at abattoirs) could further reduce the risk of contamination of meat products. © 2017 Australian Veterinary Association.

  9. [A nutrient medium for the isolation and cultivation of Campylobacter].

    Science.gov (United States)

    Temirkhanova, Z U; Gashimova, P Sh; Safonova, N V; Moroz, A F; Khazenson, L V

    1999-01-01

    Campylobacter agar, nutrient medium intended for the isolation of bacteria of the genus Campylobacter from clinical material, has been developed. The composition of the medium includes sprat hydrolysate, aerotolerant additive (ferric sulfate--oxide, sodium pyruvate, sodium pyrosulfite), sodium glutaminate, agar. The selective properties of the medium are ensured by introducing the mixture of antibiotics consisting of polymyxin B, rifampicin, amphotericin B, ristomycin. The balanced composition of Campylobacter agar ensures the aerotolerance of Campylobacter organisms and gives the optimal conditions for their growth when the inoculated material is cultivated in the atmosphere made up of the mixture of three gases (5% of oxygen, 10% of carbon dioxide, 85% of nitrogen), as well as under the conditions of a "candle vessel". The medium suppresses the development of the associative microflora diluted 10(-1). As shown in the trial of the quality of Campylobacter agar by the inoculation of material taken from patients with acute enteric infections, agricultural animals and monkeys, the medium has pronounced selective, properties with regard to extraneous microflora, while ensuring the isolation of Campylobacter on the level of the control medium.

  10. Phylogenetic diversity and position of the genus Campylobacter

    Science.gov (United States)

    Lau, P. P.; DeBrunner-Vossbrinck, B.; Dunn, B.; Miotto, K.; MacDonnell, M. T.; Rollins, D. M.; Pillidge, C. J.; Hespell, R. B.; Colwell, R. R.; Sogin, M. L.; hide

    1987-01-01

    RNA sequence analysis has been used to examine the phylogenetic position and structure of the genus Campylobacter. A complete 5S rRNA sequence was determined for two strains of Campylobacter jejuni and extensive partial sequences of the 16S rRNA were obtained for several strains of C. jejuni and Wolinella succinogenes. In addition limited partial sequence data were obtained from the 16S rRNAs of isolates of C. coli, C. laridis, C. fetus, C. fecalis, and C. pyloridis. It was found that W. succinogenes is specifically related to, but not included, in the genus Campylobacter as presently constituted. Within the genus significant diversity was noted. C. jejuni, C. coli and C. laridis are very closely related but the other species are distinctly different from one another. C. pyloridis is without question the most divergent of the Campylobacter isolates examined here and is sufficiently distinct to warrant inclusion in a separate genus. In terms of overall position in bacterial phylogeny, the Campylobacter/Wolinella cluster represents a deep branching most probably located within an expanded version of the Division containing the purple photosynthetic bacteria and their relatives. The Campylobacter/Wolinella cluster is not specifically includable in either the alpha, beta or gamma subdivisions of the purple bacteria.

  11. 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

  12. Antibiotic resistance trends and mechanisms in the foodborne pathogen, Campylobacter.

    Science.gov (United States)

    Tang, Yizhi; Fang, Liangxing; Xu, Changyun; Zhang, Qijing

    2017-11-23

    Campylobacter is a major foodborne pathogen and is commonly present in food producing animals. This pathogenic organism is highly adaptable and has become increasingly resistant to various antibiotics. Recently, both the Centers for Disease Control and Prevention and the World Health Organization have designated antibiotic-resistant Campylobacter as a serious threat to public health. For the past decade, multiple mechanisms conferring resistance to clinically important antibiotics have been described in Campylobacter, and new resistance mechanisms constantly emerge in the pathogen. Some of the recent examples include the erm(B) gene conferring macrolide resistance, the cfr(C) genes mediating resistance to florfenicol and other antimicrobials, and a functionally enhanced variant of the multidrug resistance efflux pump, CmeABC. The continued emergence of new resistance mechanisms illustrates the extraordinary adaptability of Campylobacter to antibiotic selection pressure and demonstrate the need for innovative strategies to control antibiotic-resistant Campylobacter. In this review, we will briefly summarize the trends of antibiotic resistance in Campylobacter and discuss the mechanisms of resistance to antibiotics used for animal production and important for clinical therapy in humans. A special emphasis will be given to the newly discovered antibiotic resistance.

  13. Detection of Campylobacter in human faecal samples in Fiji.

    Science.gov (United States)

    Devi, Aruna; Wilkinson, Jenny; Mahony, Timothy; Vanniasinkam, Thiru

    2014-01-01

    Data on campylobacteriosis in developed countries are well documented; in contrast, few studies on campylobacteriosis have been conducted in developing countries. This study was undertaken to test for Campylobacter in human faecal samples sent to the two major pathology laboratories in Fiji. A total of 408 diarrhoeal faecal samples were collected from the two major hospital pathology laboratories in Central Fiji (Suva) and Western Fiji (Lautoka) between December 2012 and February 2013 and from June to July 2013. Samples were analysed for the presence of Campylobacter using polymerase chain reaction (PCR) based methods. Campylobacter was detected in 241/408 (59.1%) of samples tested using PCR. Samples from children aged less than five accounted for 21.6% of positive cases. Campylobacter was detected in 59.1% of diarrhoeal samples collected from the two main laboratories in Fiji. A high proportion of children under five years with Campylobacter has been reported in other countries and could be due to parents being more likely to seek medical attention. Further studies are required to confirm the species of Campylobacter that are predominantly associated with gastroenteritis in Fiji.

  14. Detection of Campylobacter in human faecal samples in Fiji

    Directory of Open Access Journals (Sweden)

    Aruna Devi

    2014-12-01

    Full Text Available Introduction: Data on campylobacteriosis in developed countries are well documented; in contrast, few studies on campylobacteriosis have been conducted in developing countries. This study was undertaken to test for Campylobacter in human faecal samples sent to the two major pathology laboratories in Fiji. Methods: A total of 408 diarrhoeal faecal samples were collected from the two major hospital pathology laboratories in Central Fiji (Suva and Western Fiji (Lautoka between December 2012 and February 2013 and from June to July 2013. Samples were analysed for the presence of Campylobacter using polymerase chain reaction (PCR based methods. Results: Campylobacter was detected in 241/408 (59.1% of samples tested using PCR. Samples from children aged less than five accounted for 21.6% of positive cases. Discussion: Campylobacter was detected in 59.1% of diarrhoeal samples collected from the two main laboratories in Fiji. A high proportion of children under five years with Campylobacter has been reported in other countries and could be due to parents being more likely to seek medical attention. Further studies are required to confirm the species of Campylobacter that are predominantly associated with gastroenteritis in Fiji.

  15. 76 FR 15282 - New Performance Standards for Salmonella and Campylobacter in Young Chicken and Turkey Slaughter...

    Science.gov (United States)

    2011-03-21

    ... Performance Standards for Salmonella and Campylobacter in Young Chicken and Turkey Slaughter Establishments... young chicken (broiler) and turkey slaughter establishments. The new performance standards were... Salmonella and new Campylobacter performance standards for young chickens and turkeys will take effect with...

  16. A comparison of molecular technologies and genomotyping for tracing and strain characterization of Campylobacter isolates

    NARCIS (Netherlands)

    Vossen, J. van der; Keijser, B.; Schuren, F.; Nocker, A.; Montijn, R.

    2011-01-01

    Thermophilic Campylobacter species are the most common cause of gastroenteritis in developed countries. Campylobacter spp. are ubiquitous in nature and widespread in livestock. Infections occur sporadically, and are believed to occur through consumption of contaminated meat products and from

  17. Characterization and subgrouping of Campylobacter concisus strains using protein profiles, conventional biochemical testing and antibiotic susceptibility

    DEFF Research Database (Denmark)

    Aabenhus, Rune Munck; Permin, Henrik; Andersen, Leif P

    2005-01-01

    To characterize and subgroup clinical strains of Campylobacter concisus isolated from patients with gastrointestinal disease.......To characterize and subgroup clinical strains of Campylobacter concisus isolated from patients with gastrointestinal disease....

  18. Evaluation of PCR for detection of Campylobacter in a national broiler surveillance programme in Denmark

    DEFF Research Database (Denmark)

    Lund, Marianne; Wedderkopp, A; Wainø, M

    2003-01-01

    To develop and evaluate a rapid and sensitive PCR method for detection of Campylobacter spp. directly from chicken faeces.......To develop and evaluate a rapid and sensitive PCR method for detection of Campylobacter spp. directly from chicken faeces....

  19. Mg2+ improves the thermotolerance of probiotic Lactobacillus rhamnosus GG, Lactobacillus casei Zhang and Lactobacillus plantarum P-8.

    Science.gov (United States)

    Yang, Y; Huang, S; Wang, J; Jan, G; Jeantet, R; Chen, X D

    2017-04-01

    Food-related carbohydrates and proteins are often used as thermoprotectants for probiotic lactobacilli during industrial production and processing. However, the effect of inorganic salts is rarely reported. Magnesium is the second-most abundant cation in bacteria, and commonly found in various foods. Mg 2+ homeostasis is important in Salmonella and has been reported to play a critical role in their thermotolerance. However, the role of Mg 2+ in thermotolerance of other bacteria, in particular probiotic bacteria, still remains a hypothesis. In this study, the effect of Mg 2+ on thermotolerance of probiotic lactobacilli was investigated in three well-documented probiotic strains, Lactobacillus rhamnosus GG, Lactobacillus casei Zhang and Lactobacillus plantarum P-8, in comparison with Zn 2+ and Na + . Concentrations of Mg 2+ between 10 and 50 mmol l -1 were found to increase the bacterial survival upon heat challenge. Remarkably, Mg 2+ addition at 20 mmol l -1 led to a 100-fold higher survival of L. rhamnosus GG upon heat challenge. This preliminary study also showed that Mg 2+ shortened the heat-induced extended lag time of bacteria, which indicated the improvement in bacterial recovery from thermal injury. In order to improve the productivity and stability of live probiotics, extensive investigations have been carried out to improve thermotolerance of probiotics. However, most of these studies focused on the effects of carbohydrates, proteins or amino acids. The roles of inorganic salts in various food materials, which have rarely been reported, should be considered when incorporating probiotics into these foods. In this study, Mg 2+ was found to play a significant role in the thermotolerance of probiotic lactobacilli. A novel strategy may be available in the near future by employing magnesium salts as protective agents of probiotics during manufacturing process. © 2017 The Society for Applied Microbiology.

  20. 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%).

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

    Directory of Open Access Journals (Sweden)

    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.

  2. LlHSFA1, a novel heat stress transcription factor in lily (Lilium longiflorum), can interact with LlHSFA2 and enhance the thermotolerance of transgenic Arabidopsis thaliana.

    Science.gov (United States)

    Gong, Benhe; Yi, Jin; Wu, Jian; Sui, Juanjuan; Khan, Muhammad Ali; Wu, Ze; Zhong, Xionghui; Seng, Shanshan; He, Junna; Yi, Mingfang

    2014-09-01

    A heat stress transcription factor LlHSFA1 in lily and its relationship with LlHSFA2 was investigated, and its function in enhancing thermotolerance was confirmed by analyzing transgenic Arabidopsis thaliana overexpressed LlHSFA1. A large family of heat stress transcription factors that are involved in the heat stress response in plants can induce the expression of multiple genes related to thermotolerance including heat-shock proteins. In this study, a novel class A1 HSF named LlHSFA1 was isolated from leaves of lily (Lilium longiflorum cv. 'White Heaven') using the rapid amplification of cDNA ends technique. Analysis of the deduced amino acid sequence and construction of a phylogenetic tree showed that LlHSFA1 contained five critical domains and motifs and belonged to the A1 family of HSFs. Following the heat treatment of lily leaves, transcription of LlHSFA1 was induced to a varying extent, related to the time of measurement. The induced expression peak of LlHSFA1 occurred prior to that of LlHSFA2, during the early phase of heat stress. Following transient expression of LlHSFA1 in Nicotiana benthamiana, LlHSFA1 was found to be localized in both the nucleus and the cytoplasm. Analysis using bimolecular fluorescence complementation and a yeast two-hybrid assay demonstrated that LlHSFA1 could interact with LlHSFA2. Use of a yeast one-hybrid assay confirmed that LlHSFA1 had transcriptional activation activity. In transgenic Arabidopsis lines overexpressing LlHSFA1 under unstressed conditions, the expression of some putative target genes was up-regulated, in comparison with expression in wild-type plants, and furthermore, the thermotolerance of the transgenic lines was enhanced. Overall, LlHSFA1 was demonstrated to play an important role in the heat stress response of lily and to be a novel candidate gene for application in lily breeding, using genetic modification approaches.

  3. 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

  4. An efficient thermotolerant and halophilic biosurfactant-producing bacterium isolated from Dagang oil field for MEOR application

    Science.gov (United States)

    Wu, Langping; Richnow, Hans; Yao, Jun; Jain, Anil

    2014-05-01

    Dagang Oil field (Petro China Company Limited) is one of the most productive oil fields in China. In this study, 34 biosurfactant-producing strains were isolated and cultured from petroleum reservoir of Dagang oil field, using haemolytic assay and the qualitative oil-displacement test. On the basis of 16S rDNA analysis, the isolates were closely related to the species in genus Pseudomonas, Staphylococcus and Bacillus. One of the isolates identified as Bacillus subtilis BS2 were selected for further study. This bacterium was able to produce a type of biosurfactant with excessive foam-forming properties at 37ºC as well as at higher temperature of 55ºC. The biosurfactant produced by the strain BS2 could reduce the surface tension of the culture broth from 70.87 mN/m to 28.97 mN/m after 8 days of incubation at 37ºC and to 36.15 mN/m after 20 days of incubation at 55ºC, respectively. The biosurfactant showed stability at high temperature (up to 120ºC), a wide range of pH (2 to 12) and salt concentrations (up to 12%) offering potential for biotechnology. Fourier transform infrared (FT-IR) spectrum of extracted biosurfactant tentatively characterized the produced biosurfactant as glycolipid derivative. Elemental analysis of the biosurfactant by energy dispersive X-ray spectroscopy (EDS) reveals that the biosurfactant was anionic in nature. 15 days of biodegradation of crude oil suggested a preferential usage of n-alkane upon microbial metabolism of BS2 as a carbon substrate and consequently also for the synthesis of biosurfactants. Core flood studies for oil release indicated 9.6% of additional oil recovery over water flooding at 37ºC and 7.2% of additional oil recovery at 55 ºC. Strain BS2 was characterized as an efficient biosurfactant-producing, thermotolerant and halophillic bacterium and has the potential for application for microbial enhanced oil recovery (MEOR) through water flooding in China's oil fields even in situ as adapted to reservoir chemistry and

  5. Impact of technical and economic performance on costs of campylobacter spp. interventions on broiler farms in six European countries

    NARCIS (Netherlands)

    Wagenberg, van C.P.A.; Horne, van P.L.M.

    2016-01-01

    Campylobacter spp. is one of the leading causes of acute diarrheal disease in humans worldwide and human Campylobacter spp. infections can result in severe sequelae. Because broilers are an important reservoir for human Campylobacter spp. infections, it is relevant to control Campylobacter spp.

  6. Detection of Campylobacter species and Arcobacter butzleri in stool samples by use of real-time multiplex PCR

    NARCIS (Netherlands)

    R.F. de Boer (Richard); A. Ott (Alewijn); P. Güren (Pinar); E. van Zanten; A.F. van Belkum (Alex); A.M.D. Kooistra-Smid

    2013-01-01

    textabstractThe presence of Campylobacter (or Campylobacter-like) species in stools from patients suspected of infectious gastroenteritis (n = 493) was investigated using real-time PCR for detection of Arcobacter butzleri (hsp60 gene), Campylobacter coli (ceuE gene), Campylobacter jejuni (mapA),

  7. Hormone assay

    International Nuclear Information System (INIS)

    Eisentraut, A.M.

    1977-01-01

    An improved radioimmunoassay is described for measuring total triiodothyronine or total thyroxine levels in a sample of serum containing free endogenous thyroid hormone and endogenous thyroid hormone bound to thyroid hormone binding protein. The thyroid hormone is released from the protein by adding hydrochloric acid to the serum. The pH of the separated thyroid hormone and thyroid hormone binding protein is raised in the absence of a blocking agent without interference from the endogenous protein. 125 I-labelled thyroid hormone and thyroid hormone antibodies are added to the mixture, allowing the labelled and unlabelled thyroid hormone and the thyroid hormone antibody to bind competitively. This results in free thyroid hormone being separated from antibody bound thyroid hormone and thus the unknown quantity of thyroid hormone may be determined. A thyroid hormone test assay kit is described for this radioimmunoassay. It provides a 'single tube' assay which does not require blocking agents for endogenous protein interference nor an external solid phase sorption step for the separation of bound and free hormone after the competitive binding step; it also requires a minimum number of manipulative steps. Examples of the assay are given to illustrate the reproducibility, linearity and specificity of the assay. (UK)

  8. Analysis of the baseline survey on the prevalence of Campylobacter in broiler batches and of Campylobacter and Salmonella on broiler carcasses in the EU, 2008, Part A: Campylobacter and Salmonella prevalence estimates

    DEFF Research Database (Denmark)

    Hald, Tine

    A European Union-wide baseline survey on Campylobacter in broiler batches and on Campylobacter and Salmonella on broiler carcasses was carried out in 2008. A total of 10,132 broiler batches were sampled from 561 slaughterhouses in 26 European Union Member States and two countries not belonging......-contaminated broiler carcasses was 75.8%. The Member State prevalence varied from 2.0% to 100.0% and from 4.9% to 100.0%, for caecal contents and carcasses, respectively. The results of the counts of Campylobacter on broiler carcasses showed substantial variation among the countries in contamination levels. About two......-thirds of the Campylobacter isolates from the pooled caecal contents as well as from the broiler carcasses were identified as Campylobacter jejuni, while one-third was Campylobacter coli. Twenty-two Member States and one non-Member State isolated Salmonella on the broiler carcasses, with a Community prevalence of 15...

  9. Campylobacter epidemiology from breeders to their progeny in Eastern Spain.

    Science.gov (United States)

    Ingresa-Capaccioni, S; Jiménez-Trigos, E; Marco-Jiménez, F; Catalá, P; Vega, S; Marin, C

    2016-03-01

    While horizontal transmission is a route clearly linked to the spread of Campylobacter at the farm level, few studies support the transmission of Campylobacter spp. from breeder flocks to their offspring. Thus, the present study was carried out to investigate the possibility of vertical transmission. Breeders were monitored from the time of housing day-old chicks, then throughout the laying period (0 to 60 wk) and throughout their progeny (broiler fattening, 1 to 42 d) until slaughter. All samples were analyzed according with official method ISO 10272:2006. Results revealed that on breeder farms, Campylobacter isolation started from wk 16 and reached its peak at wk 26, with 57.0% and 93.2% of positive birds, respectively. After this point, the rate of positive birds decreased slightly to 86.0% at 60 wk. However, in broiler production all day-old chicks were found negative for Campylobacter spp, and the bacteria was first isolated at d 14 of age (5.0%), with a significant increase in detection during the fattening period with 62% of Campylobacter positive animals at the end of the production cycle. Moreover, non-positive sample was determined from environmental sources. These results could be explained because Campylobacter may be in a low concentration or in a non-culturable form, as there were several studies that successfully detected Campylobacter DNA, but failed to culture. This form can survive in the environment and infect successive flocks; consequently, further studies are needed to develop more modern, practical, cost-effective and suitable techniques for routine diagnosis. © 2016 Poultry Science Association Inc.

  10. Understanding the mechanisms of ATPase beta family genes for cellular thermotolerance in crossbred bulls.

    Science.gov (United States)

    Deb, Rajib; Sajjanar, Basavaraj; Singh, Umesh; Alex, Rani; Raja, T V; Alyethodi, Rafeeque R; Kumar, Sushil; Sengar, Gyanendra; Sharma, Sheetal; Singh, Rani; Prakash, B

    2015-12-01

    Na+/K+-ATPase is an integral membrane protein composed of a large catalytic subunit (alpha), a smaller glycoprotein subunit (beta), and gamma subunit. The beta subunit is essential for ion recognition as well as maintenance of the membrane integrity. Present study was aimed to analyze the expression pattern of ATPase beta subunit genes (ATPase B1, ATPase B2, and ATPase B3) among the crossbred bulls under different ambient temperatures (20-44 °C). The present study was also aimed to look into the relationship of HSP70 with the ATPase beta family genes. Our results demonstrated that among beta family genes, transcript abundance of ATPase B1 and ATPase B2 is significantly (P ATPase Β1, ATPase B2, and ATPase B3 is highly correlated (P ATPase beta family genes for cellular thermotolerance in cattle.

  11. High alcohol production by solid substrate fermentation from starchy substrates using thermotolerant Saccharomyces cerevisiae

    Energy Technology Data Exchange (ETDEWEB)

    Sree, N.K.; Sridhar, M.; Suresh, K.; Rao, L.V. [Department of Microbiology, Osmania University, Hyderabad 500007, Andhra Pradesh (India)

    1999-06-01

    Solid Substrate Fermentation system (SSF) was used to produce ethanol from various starchy substrates like sweet sorghum, sweet potato, wheat flour, rice starch, soluble starch and potato starch using thermotolerant yeast isolate (VS{sub 3}) by simultaneous saccharification and fermentation process. Alcohol produced was estimated by gas chromatography after an incubation time of 96 hrs at 37 C and 42 C. More ethanol was produced from rice starch and sweet sorghum. The maximum amount of ethanol produced from these substrates using VS{sub 3} was 10 g/100 g and 3.5 g/100 g substrate (rice starch) and 8.2 g and 7.5 g/100 g substrate (sweet sorghum) at 37 C and 42 C respectively. (orig.) With 2 figs., 1 tab., 12 refs.

  12. Screening and application of thermotolerant microorganisms and their flocculant for treatment of palm oil mill effluent

    Directory of Open Access Journals (Sweden)

    Saithong Kaewchai

    2002-07-01

    Full Text Available Among fifteen thermotolerant polymer-producing isolates, three strains SM 29, WD 90, and SM 38 produced polymer posessing very high flocculating activities (24.81, 14.63 and 10.84, respectively and flocculation rates (94.29, 90.69 and 87.84, respectively. These three strains were identified to be Bacillus subtilis WD90, Bacillus subtilis SM 29, and Enterobacter agglomerans SM 38. Treatment of palm oil mill effluent (POME by these three selected strains under aerobic condition at 45ºC for 48 h revealed that neither oil separation nor flocculation of solids was observed. However, all three strains were able to decolorize the POME from dark brown to very light yellow. Flocculant produced from the three selected isolates could not separate the suspended solids and oil from the POME.

  13. A simplified and cost-effective enrichment protocol for the isolation of Campylobacter spp. from retail broiler meat without microaerobic incubation

    Science.gov (United States)

    2011-01-01

    Background To simplify the methodology for the isolation of Campylobacter spp. from retail broiler meat, we evaluated 108 samples (breasts and thighs) using an unpaired sample design. The enrichment broths were incubated under aerobic conditions (subsamples A) and for comparison under microaerobic conditions (subsamples M) as recommended by current reference protocols. Sensors were used to measure the dissolved oxygen (DO) in the broth and the percentage of oxygen (O2) in the head space of the bags used for enrichment. Campylobacter isolates were identified with multiplex PCR assays and typed using pulsed-field gel electrophoresis (PFGE). Ribosomal intergenic spacer analyses (RISA) and denaturing gradient gel electrophoresis (DGGE) were used to study the bacterial communities of subsamples M and A after 48 h enrichment. Results The number of Campylobacter positive subsamples were similar for A and M when all samples were combined (P = 0.81) and when samples were analyzed by product (breast: P = 0.75; thigh: P = 1.00). Oxygen sensors showed that DO values in the broth were around 6 ppm and O2 values in the head space were 14-16% throughout incubation. PFGE demonstrated high genomic similarity of isolates in the majority of the samples in which isolates were obtained from subsamples A and M. RISA and DGGE results showed a large variability in the bacterial populations that could be attributed to sample-to-sample variations and not enrichment conditions (aerobic or microaerobic). These data also suggested that current sampling protocols are not optimized to determine the true number of Campylobacter positive samples in retail boiler meat. Conclusions Decreased DO in enrichment broths is naturally achieved. This simplified, cost-effective enrichment protocol with aerobic incubation could be incorporated into reference methods for the isolation of Campylobacter spp. from retail broiler meat. PMID:21812946

  14. A simplified and cost-effective enrichment protocol for the isolation of Campylobacter spp. from retail broiler meat without microaerobic incubation

    LENUS (Irish Health Repository)

    Zhou, Ping

    2011-08-03

    Abstract Background To simplify the methodology for the isolation of Campylobacter spp. from retail broiler meat, we evaluated 108 samples (breasts and thighs) using an unpaired sample design. The enrichment broths were incubated under aerobic conditions (subsamples A) and for comparison under microaerobic conditions (subsamples M) as recommended by current reference protocols. Sensors were used to measure the dissolved oxygen (DO) in the broth and the percentage of oxygen (O2) in the head space of the bags used for enrichment. Campylobacter isolates were identified with multiplex PCR assays and typed using pulsed-field gel electrophoresis (PFGE). Ribosomal intergenic spacer analyses (RISA) and denaturing gradient gel electrophoresis (DGGE) were used to study the bacterial communities of subsamples M and A after 48 h enrichment. Results The number of Campylobacter positive subsamples were similar for A and M when all samples were combined (P = 0.81) and when samples were analyzed by product (breast: P = 0.75; thigh: P = 1.00). Oxygen sensors showed that DO values in the broth were around 6 ppm and O2 values in the head space were 14-16% throughout incubation. PFGE demonstrated high genomic similarity of isolates in the majority of the samples in which isolates were obtained from subsamples A and M. RISA and DGGE results showed a large variability in the bacterial populations that could be attributed to sample-to-sample variations and not enrichment conditions (aerobic or microaerobic). These data also suggested that current sampling protocols are not optimized to determine the true number of Campylobacter positive samples in retail boiler meat. Conclusions Decreased DO in enrichment broths is naturally achieved. This simplified, cost-effective enrichment protocol with aerobic incubation could be incorporated into reference methods for the isolation of Campylobacter spp. from retail broiler meat.

  15. Tuning Chocolate Flavor through Development of Thermotolerant Saccharomyces cerevisiae Starter Cultures with Increased Acetate Ester Production.

    Science.gov (United States)

    Meersman, Esther; Steensels, Jan; Struyf, Nore; Paulus, Tinneke; Saels, Veerle; Mathawan, Melissa; Allegaert, Leen; Vrancken, Gino; Verstrepen, Kevin J

    2016-01-15

    Microbial starter cultures have extensively been used to enhance the consistency and efficiency of industrial fermentations. Despite the advantages of such controlled fermentations, the fermentation involved in the production of chocolate is still a spontaneous process that relies on the natural microbiota at cocoa farms. However, recent studies indicate that certain thermotolerant Saccharomyces cerevisiae cultures can be used as starter cultures for cocoa pulp fermentation. In this study, we investigate the potential of specifically developed starter cultures to modulate chocolate aroma. Specifically, we developed several new S. cerevisiae hybrids that combine thermotolerance and efficient cocoa pulp fermentation with a high production of volatile flavor-active esters. In addition, we investigated the potential of two strains of two non-Saccharomyces species that produce very large amounts of fruity esters (Pichia kluyveri and Cyberlindnera fabianii) to modulate chocolate aroma. Gas chromatography-mass spectrometry (GC-MS) analysis of the cocoa liquor revealed an increased concentration of various flavor-active esters and a decrease in spoilage-related off-flavors in batches inoculated with S. cerevisiae starter cultures and, to a lesser extent, in batches inoculated with P. kluyveri and Cyb. fabianii. Additionally, GC-MS analysis of chocolate samples revealed that while most short-chain esters evaporated during conching, longer and more-fat-soluble ethyl and acetate esters, such as ethyl octanoate, phenylethyl acetate, ethyl phenylacetate, ethyl decanoate, and ethyl dodecanoate, remained almost unaffected. Sensory analysis by an expert panel confirmed significant differences in the aromas of chocolates produced with different starter cultures. Together, these results show that the selection of different yeast cultures opens novel avenues for modulating chocolate flavor. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

  16. Screening of thermotolerant microorganisms and application for oil separation from palm oil mill wastewater

    Directory of Open Access Journals (Sweden)

    Aran H-Kittikun

    2007-05-01

    Full Text Available The characteristics of palm oil mill wastewater (POMW were brown color, pH 3.8-4.3, temperature 48-55oC, total solids 68.2-82.1 g/l, suspended solids 26.2-65.6 g/l, oil and grease 19.1-25.1 g/l, COD 49.9-160.7g/l and BOD 32.5-75.3 g/l. After centrifugation (3,184 xg of 50 ml POMW for 10 min, the POMW was separated into 3 layers: top (oil, middle (supernatant and bottom layer (sediment. The sediment containeddry weight 1.19 g and oil and grease 1.07 g. In order to release oil and grease trapped in palm fiber debris in the POMW, cellulase- and/or xylanase-enzyme-producing and thermotolerant microorganisms wereisolated. The isolates SO1 and SO2 were isolated from soil near the first anaerobic pond of the palm oil mill. They were aerobic, Gram positive, rod shaped, thermotolerant microorganisms and produced cellulase 12.11 U/ml (3 days and 7.2 U/ml (4 days, and xylanase 50.98 U/ml (4 days and 20.42 U/ml (4 days, respectivelyin synthetic medium containing carboxymethycellulose as a carbon source. When these 2 isolates were added into the steriled POMW under shaking condition for 7 days, after centrifugation at 3,184 xg the isolate SO1gave the better % reduction of dry weight (64.66 % and of oil and grease in the bottom layer (85.32 % of the POMW.

  17. Tuning Chocolate Flavor through Development of Thermotolerant Saccharomyces cerevisiae Starter Cultures with Increased Acetate Ester Production

    Science.gov (United States)

    Meersman, Esther; Steensels, Jan; Struyf, Nore; Paulus, Tinneke; Saels, Veerle; Mathawan, Melissa; Allegaert, Leen; Vrancken, Gino

    2015-01-01

    Microbial starter cultures have extensively been used to enhance the consistency and efficiency of industrial fermentations. Despite the advantages of such controlled fermentations, the fermentation involved in the production of chocolate is still a spontaneous process that relies on the natural microbiota at cocoa farms. However, recent studies indicate that certain thermotolerant Saccharomyces cerevisiae cultures can be used as starter cultures for cocoa pulp fermentation. In this study, we investigate the potential of specifically developed starter cultures to modulate chocolate aroma. Specifically, we developed several new S. cerevisiae hybrids that combine thermotolerance and efficient cocoa pulp fermentation with a high production of volatile flavor-active esters. In addition, we investigated the potential of two strains of two non-Saccharomyces species that produce very large amounts of fruity esters (Pichia kluyveri and Cyberlindnera fabianii) to modulate chocolate aroma. Gas chromatography-mass spectrometry (GC-MS) analysis of the cocoa liquor revealed an increased concentration of various flavor-active esters and a decrease in spoilage-related off-flavors in batches inoculated with S. cerevisiae starter cultures and, to a lesser extent, in batches inoculated with P. kluyveri and Cyb. fabianii. Additionally, GC-MS analysis of chocolate samples revealed that while most short-chain esters evaporated during conching, longer and more-fat-soluble ethyl and acetate esters, such as ethyl octanoate, phenylethyl acetate, ethyl phenylacetate, ethyl decanoate, and ethyl dodecanoate, remained almost unaffected. Sensory analysis by an expert panel confirmed significant differences in the aromas of chocolates produced with different starter cultures. Together, these results show that the selection of different yeast cultures opens novel avenues for modulating chocolate flavor. PMID:26590272

  18. Evaluation of Aegilops tauschii and Aegilops speltoides for acquired thermotolerance: Implications in wheat breeding programmes.

    Science.gov (United States)

    Hairat, Suboot; Khurana, Paramjit

    2015-10-01

    Severe and frequent heat waves are predicted in the near future having dramatic and far-reaching ecological and social impact. The aim of this study was to examine acquired thermotolerance of two Aegilops species: Aegilops tauschii and Aegilops speltoides and study their potential adaptive mechanisms. The effect of two episodes of high heat stress (45 °C/12 h) with a day of recovery period was investigated on their physiology. As compared to A. speltoides, A. tauschii suffered less inhibition of photosystem II efficiency and net photosynthetic rate (Pn). Although A. tauschii showed nearly complete recovery of PSII, the adverse effect was more pronounced in A. speltoides. Measurement of the minimum fluorescence (Fo) versus temperature curves revealed a higher inflection temperature of Fo for A. tauschii than A. speltoides, reflecting greater thermo stability of the photosynthetic apparatus. Absorbed light energy distribution revealed that A. speltoides showed increased steady state fluorescence and a lower absorbed light allocated to photosynthetic chemistry (ɸPSII) relative to A. tauschii. However, A. tauschii showed higher ability to scavenge free radicals as compared to A. speltoides. This was further validated by higher expression of ascorbate peroxidase gene. These results suggest that A. tauschii showed faster recovery and a better thermostability of its photosynthetic apparatus under severe stress conditions along with a better regulation of energy channeling of PSII complexes to minimize oxidative damage and thus retains greater capability of carbon assimilation. These factors aid in imparting a greater heat tolerance to A. tauschii as compared to A. speltoides and thus make it a better candidate for alien species introgression in wheat breeding programs for thermotolerance in wheat. Copyright © 2015 Elsevier Masson SAS. All rights reserved.

  19. Root Antioxidant Mechanisms in Relation to Root Thermotolerance in Perennial Grass Species Contrasting in Heat Tolerance.

    Directory of Open Access Journals (Sweden)

    Yi Xu

    Full Text Available Mechanisms of plant root tolerance to high temperatures through antioxidant defense are not well understood. The objective of this study was to investigate whether superior root thermotolerance of heat-tolerant Agrostis scabra relative to its congeneric heat-sensitive Agrostis stolonifera was associated with differential accumulation of reactive oxygen species and antioxidant scavenging systems. A. scabra 'NTAS' and A. stolonifera 'Penncross' plants were exposed to heat stress (35/30°C, day/night in growth chambers for 24 d. Superoxide (O2(- content increased in both A. stolonifera and A. scabra roots under heat stress but to a far lesser extent in A. scabra than in A. stolonifera. Hydrogen peroxide (H2O2 content increased significantly in A. stolonifera roots but not in A. scabra roots responding to heat stress. The content of antioxidant compounds (ascorbate and glutathione did not differ between A. stolonifera and A. scabra under heat stress. Enzymatic activity of superoxide dismutase was less suppressed in A. scabra than that in A. stolonifera under heat stress, while peroxidase and catalase were more induced in A. scabra than in A. stolonifera. Similarly, their encoded transcript levels were either less suppressed, or more induced in A. scabra roots than those in A. stolonifera during heat stress. Roots of A. scabra exhibited greater alternative respiration rate and lower cytochrome respiration rate under heat stress, which was associated with suppression of O2(- and H2O2 production as shown by respiration inhibitors. Superior root thermotolerance of A. scabra was related to decreases in H2O2 and O2(- accumulation facilitated by active enzymatic antioxidant defense systems and the maintenance of alternative respiration, alleviating cellular damages by heat-induced oxidative stress.

  20. Effects of dry whey powder and calcium butyrate supplementation of corn/soybean-based diets on productive performance, duodenal histological integrity, and Campylobacter colonization in broilers.

    Science.gov (United States)

    Ocejo, Medelin; Oporto, Beatriz; Juste, Ramón A; Hurtado, Ana

    2017-06-26

    were observed during the assay. Beneficial effects on performance and intestinal health were observed, particularly during the starter period, when chickens were fed a diet supplemented with both whey and coated calcium butyrate. However, none of the tested diets provided the chicks any differential degree of protection against Campylobacter infection.

  1. 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...

  2. Bibliometric analysis of publications on Campylobacter: (2000-2015).

    Science.gov (United States)

    Sweileh, Waleed M; Al-Jabi, Samah W; Sawalha, Ansam F; AbuTaha, Adham S; Zyoud, Sa'ed H

    2016-11-29

    Campylobacter species are widespread zoonotic pathogens. Campylobacter jejuni causes a form of gastroenteritis called campylobacteriosis. Campylobacter drug resistance is considered a serious threat. In order to better understand national and international research output on Campylobacter, we conducted this bibliometric overview of publications on Campylobacter. This study can be used to assess extent of interaction and response of researchers, food regulators, and health policy makers to global burden of campylobacateriosis. Scopus database was used to retrieve publications with the following keywords (Campylobacter/campylobacteriosis, C. jejuni, C. coli). The study period was set from 2000 to 2015. All types of journal documents, excluding errata, were considered. Bibliometric indicators such as annual growth of publications, country contribution, international collaboration, and citation analysis were presented. The quality of retrieved data was indirectly assessed by Hirsch index and impact factor of journals. A total of 5522 documents were retrieved with median (Q1-Q3) citations of 9 (2-23) and h-index of 113. Annual number of publications showed a fluctuating increase. The core leading journals were Applied and Environmental Microbiology journal and Journal of Food Protection with 246 (4.46%) publications for each. The USA (1309; 23.6%) was the most productive country while Danmarks Tekniske Universitet (150; 2.7%) was the most productive institution. Half of the top ten productive countries were European. France had the lowest percentage (33.5%) of articles with international collaboration while Netherlands (57.7%) had the highest percentage of articles with international collaboration. Approximately half (50.1%) of retrieved articles were published in journals under the subject area of "immunology/microbiology". Main themes in highly cited articles were molecular biology/genetics and public health burden of campylobacteriosis. There were 728 (13

  3. 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.

  4. 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.

  5. Molecular subtyping and erythromycin resistance of Campylobacter in China.

    Science.gov (United States)

    Zhang, A; Song, L; Liang, H; Gu, Y; Zhang, C; Liu, X; Zhang, J; Zhang, M

    2016-07-01

    To investigate the erythromycin resistance patterns and mechanism for Campylobacter isolates in China. The minimum inhibitory concentrations of erythromycin on 858 Chinese Campylobacter isolates were analysed. PCR and DNA sequencing were used to identify mutations in the 23S rRNA and the presence of the ermB gene in the 158 erythromycin resistance isolates (18·4%). About 83% (131/158) had A2075G mutation in their 23S rRNA; no A2074C/G mutants were found. The ermB gene was identified in 30 Campylobacter coli isolates (19%). Four types of multidrug-resistant gene islands (MDRGIs) were found. Fifty-three types were identified by multilocus sequence typing among the resistant isolates. All isolates of STs 6322 and 1145 had the ermB gene. The erythromycin resistance rate of Camp. coli (58·56%) was much higher than Campylobacter jejuni (0·67%). The insertion sites between cadF and CCO1582 and between nfsB and cinA on the chromosome might be hot spots for MDRGI transformation. Point mutation in domain V of the 23S rRNA and the ermB gene accounted for 100% of the erythromycin resistance of Campylobacter in China. Journal of Applied Microbiology © 2016 The Society for Applied Microbiology.

  6. Survival with a helping hand: Campylobacter and microbiota

    Directory of Open Access Journals (Sweden)

    Ivana eIndikova

    2015-11-01

    Full Text Available Campylobacteriosis is the most important bacterial food-borne disease in the developed world. Consumption of chicken meat, beef or raw milk, direct contact with ruminants and exposure to contaminated surface water or even consumption of tap water have been identified as risk factors for human disease. However, the most important risk factor is consumption of and/or handling contaminated chicken. Campylobacter spp. are fastidious microorganisms but must somehow survive outside the host, especially in food and agricultural environments and also resist the innate and humoral immune responses inside the host. In this paper we hypothesize that other microorganisms in mixed populations with Campylobacter may act to improve survival outside the host and may also protect the pathogen against the intestinal immune system. Our evidence for this hypothesis is based on: 1. newly generated microbial community analysis; 2. the prolonged survival of Campylobacter in mixed species biofilms and in co-culture with environmental bacteria; 3. improved survival in amoebae and rumen fluid; 4. sulphur release and iron uptake systems within the intestinal lumen. This would make Campylobacter an exceptional food-borne pathogen. With this in mind, new strategies are necessary to combat Campylobacter along the total food chain.

  7. QUANTITATIVE ASSESSMENT OF CAMPYLOBACTER SPP. ON POULTRY CARCASSES

    Directory of Open Access Journals (Sweden)

    L. Alberghini

    2011-01-01

    Full Text Available Campylobacter spp. are bacterial pathogens associated with human gastroenteritis worldwide. In Europe, campylobacteriosis is one of the leading food-borne bacterial diseases and the consumption of poultry meats is suspected to be one of the major causes of illness. The aim of our research was to determine the number of Campylobacter spp. in poultry carcasses and in poultry meat samples during their storage till to retail markets. The study was conducted from February 2009 to February 2010 at slaughterhouse in Veneto region, followed by a test of fresh poultry meat placed on the market for sale. A total of 90 poultry carcass and 90 samples of poultry meat were examined. The quantitative examination resulted in Campylobacter spp. counts (mean: for carcasses between 2,0 ∙101 ufc/g and 1,5 ∙103 ufc/g (4,2 ∙102 and poultry meat between 2,0 ∙101 ufc/g and 3,7 ∙102 ufc/g (8,1 ∙101. The majority of isolates were classified as Campylobacter jejuni (58,3%, Campylobacter coli (22,9% or Arcobacter cryaerophilus (4,2%. Acknowledgments: The project was funded with grants from Fondazione Cariverona 2007.

  8. [Campylobacter and Salmonella acute gastroenteritis: epidemiology and health care utilization].

    Science.gov (United States)

    Sala Farré, Maria Rosa; Osorio Sánchez, Dimelza; Arias Varela, Cesar; Simó Sanahuja, Maria; Recasens Recasens, Assumpta; Pérez Jové, Josefa

    2015-10-05

    In Catalonia the current surveillance systems do not allow to know the true incidence or the health care utilization of acute gastroenteritis (AGE) caused by Campylobacter and Salmonella infections. The aim of this study is to analyze these characteristics. Descriptive study of Campylobacter and Salmonella infections reported in 2002 and 2012 in Catalonia, Spain. We included cases isolated and reported by the laboratory to a regional Surveillance Unit. The estimated incidence of Salmonella and Campylobacter AGE decreased by almost 50% and 20% respectively in 2012. Children between one and 4 years old were the most affected in both years. Significant differences in the clinical characteristics and disease duration were observed between Campylobacter and Salmonella. Visits to the Emergency Department and hospitalization rates were 63.7% and 15%, being more frequent among salmonellosis cases. The estimated incidence of Campylobacter and Salmonella infections has decreased, however rates are still important, as well as it is the health care utilization in both diseases. Current surveillance systems need appropriateness improvements to reach a better control of these infections. Copyright © 2014 Elsevier España, S.L.U. All rights reserved.

  9. Rapidly decreased serum IgG to Campylobacter pylori following elimination of Campylobacter in histological chronic biopsy Campylobacter-positive gastritis

    NARCIS (Netherlands)

    van Bohemen, C. G.; Langenberg, M. L.; Rauws, E. A.; Oudbier, J.; Weterings, E.; Zanen, H. C.

    1989-01-01

    The anaerobic bacterium Campylobacter pylori (Cp) is thought to be associated with chronic gastritis. This paper presents clinical data underpinning this view. Five patients with histological chronic gastritis as determined by diagnostic endoscopy, which was associated with Cp as determined by

  10. Quantitative Microbial Risk Assessment for Campylobacter spp. on Ham in Korea.

    Science.gov (United States)

    Lee, Jeeyeon; Ha, Jimyeong; Kim, Sejeong; Lee, Heeyoung; Lee, Soomin; Yoon, Yohan

    2015-01-01

    The objective of this study was to evaluate the risk of illness from Campylobacter spp. on ham. To identify the hazards of Campylobacter spp. on ham, the general characteristics and microbial criteria for Campylobacter spp., and campylobacteriosis outbreaks were investigated. In the exposure assessment, the prevalence of Campylobacter spp. on ham was evaluated, and the probabilistic distributions for the temperature of ham surfaces in retail markets and home refrigerators were prepared. In addition, the raw data from the Korea National Health and Nutrition Examination Survey (KNHNES) 2012 were used to estimate the consumption amount and frequency of ham. In the hazard characterization, the Beta-Poisson model for Campylobacter spp. infection was used. For risk characterization, a simulation model was developed using the collected data, and the risk of Campylobacter spp. on ham was estimated with @RISK. The Campylobacter spp. cell counts on ham samples were below the detection limit (ham was 23.93 g per person, and the consumption frequency was 11.57%. The simulated mean value of the initial contamination level of Campylobacter spp. on ham was -3.95 Log CFU/g, and the mean value of ham for probable risk per person per day was 2.20×10(-12). It is considered that the risk of foodborne illness for Campylobacter spp. was low. Furthermore, these results indicate that the microbial risk assessment of Campylobacter spp. in this study should be useful in providing scientific evidence to set up the criteria of Campylobacter spp..

  11. Polymerase chain reaction detection of naturally occurring Campylobacter in commercial broiler chicken embryos.

    Science.gov (United States)

    Hiett, K L; Cox, N A; Rothrock, M J

    2013-04-01

    Campylobacter, a foodborne pathogen closely associated with poultry, is recognized as a leading bacterial etiologic agent of human gastroenteritis in the United States. In this investigation, 2 trials were performed where tissues from 7-, 14/15-, and 19-d-old commercial broiler chicken embryos were tested for the presence of Campylobacter using both culturing methodology and PCR. Conventional culturing methods failed to detect Campylobacter from any samples tested during this investigation. Using a set of primers specific for the Campylobacter flagellinA short variable region (flaA SVR), Campylobacter DNA was amplified in 100, 80, and 100% of gastrointestinal tracts from 7-, 15-, and 19-d-old embryos, respectively, in the first trial. Similarly, Campylobacter DNA was detected in 100, 70, and 60% of gastrointestinal tracts of 7-, 14-, and 18-d-old embryos, respectively, in the second trial. In both trials, yolk sac, albumin, and liver/gallbladder samples from 19-d-old embryos all failed to produce amplicons indicative of Campylobacter DNA. Subsequent DNA sequence analyses of the flaA SVR PCR products were consistent with the amplicon arising from Campylobacter. Although a determination of whether the Campylobacter was living or dead within the embryos could not be made, these results demonstrate that Campylobacter-specific DNA is present within the gastrointestinal tract of broiler chicken embryos; however, the means by which it is present and the relative contribution to subsequent Campylobacter contamination of poultry flocks requires further investigation.

  12. 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

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

    Directory of Open Access Journals (Sweden)

    Sunyoung Hwang

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

  14. Campylobacter ornithocola sp. nov., a novel member of the Campylobacter lari group isolated from wild bird faecal samples.

    Science.gov (United States)

    Cáceres, Alberto; Muñoz, Ivo; Iraola, Gregorio; Díaz-Viraqué, Florencia; Collado, Luis

    2017-06-01

    During a study on the prevalence and diversity of campylobacteria in wild birds faecal samples from the city of Valdivia (southern Chile) 17 Gram-stain-negative, curved-rod-shaped isolates, were initially identified as Campylobacter lari by PCR-RFLP. Further identification by 16S rRNA sequence analysis revealed that they formed a distinct group in the genus Campylobacter. This unique position was confirmed by the results of analysis of rpoB, atpA and cpn60 gene sequences. The average nucleotide identity between the representative strain WBE38T and the type strain of the most closely related taxon C. larisubsp.concheus (LMG 11760) was 90.8 %. The oxidase and urease activity of the novel isolates enabled them to be phenotypically differentiated from species of the genus Campylobacter with validly published names. Therefore, on the basis of phenotypic, genetic and genomic characterizations, the results of this study clearly indicate that these strains represent a novel species within the genus Campylobacter, for which the name Campylobacter ornithocola sp. nov. is proposed, with the type strain WBE38T (=CECT 9147T=LMG 29815T).

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

    Directory of Open Access Journals (Sweden)

    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.

  16. 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.

  17. 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.

  18. Production of pullulan by a thermotolerant aureobasidium pullulans strain in non-stirred fed batch fermentation process.

    Science.gov (United States)

    Singh, Ranjan; Gaur, Rajeeva; Tiwari, Soni; Gaur, Manogya Kumar

    2012-07-01

    Total 95 isolates of Aureobasidium pullulans were isolated from different flowers and leaves samples, out of which 11 thermotolerant strains produced pullulan. One thermotolerant non-melanin pullulan producing strain, designated as RG-5, produced highest pullulan (37.1±1.0 g/l) at 42(o)C, pH 5.5 in 48h of incubation with 3% sucrose and 0.5% ammonium sulphate in a non-stirred fed batch fermentor of 6 liters capacity. The two liters of initial volume of fermentation medium was further fed with the 2 liters in two successive batches at 5 h interval into the fermentor. The sterile air was supplied only for 10h at the rate of 0.5 vvm.

  19. Production of pullulan by a thermotolerant Aureobasidium pullulans strain in non-stirred fed batch fermentation process

    Directory of Open Access Journals (Sweden)

    Ranjan Singh

    2012-09-01

    Full Text Available Total 95 isolates of Aureobasidium pullulans were isolated from different flowers and leaves samples, out of which 11 thermotolerant strains produced pullulan. One thermotolerant non-melanin pullulan producing strain, designated as RG-5, produced highest pullulan (37.1±1.0 g/l at 42ºC, pH 5.5 in 48h of incubation with 3% sucrose and 0.5% ammonium sulphate in a non-stirred fed batch fermentor of 6 liters capacity. The two liters of initial volume of fermentation medium was further fed with the 2 liters in two successive batches at 5 h interval into the fermentor. The sterile air was supplied only for 10h at the rate of 0.5 vvm.

  20. Campylobacter pinnipediorum sp. nov., isolated from pinnipeds, comprising Campylobacter pinnipediorum subsp. pinnipediorum subsp. nov. and Campylobacter pinnipediorum subsp. caledonicus subsp. nov.

    Science.gov (United States)

    Gilbert, Maarten J; Miller, William G; Leger, Judy St; Chapman, Mary H; Timmerman, Arjen J; Duim, Birgitta; Foster, Geoffrey; Wagenaar, Jaap A

    2017-06-01

    During independent diagnostic screenings of otariid seals in California (USA) and phocid seals in Scotland (UK), Campylobacter-like isolates, which differed from the established taxa of the genus Campylobacter, were cultured from abscesses and internal organs of different seal species. A polyphasic study was undertaken to determine the taxonomic position of these six isolates. The isolates were characterized by 16S rRNA gene and AtpA sequence analysis and by conventional phenotypic testing. The whole-genome sequences were determined for all isolates, and the average nucleotide identity (ANI) was determined. The isolates formed a separate phylogenetic clade, divergent from all other taxa of the genus Campylobacter and most closely related to Campylobactermucosalis. Although all isolates showed 100 % 16S rRNA gene sequence homology, AtpA and ANI analyses indicated divergence between the otariid isolates from California and the phocid isolates from Scotland, which warrants subspecies status for each clade. The two subspecies could also be distinguished phenotypically on the basis of catalase activity. This study shows clearly that the isolates obtained from pinnipeds represent a novel species within the genus Campylobacter, for which the name Campylobacter pinnipediorum sp. nov. is proposed. Within this novel species, the Californian isolates represent a separate subspecies, for which the name C. pinnipediorum subsp. pinnipediorum subsp. nov. is proposed. The type strain for both this novel species and subspecies is RM17260T (=LMG 29472T=CCUG 69570T). The Scottish isolates represent another subspecies, for which the name C. pinnipediorum subsp. caledonicus subsp. nov. is proposed. The type strain of this subspecies is M302/10/6T (=LMG 29473T=CCUG 68650T).