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multi-locus sequence types of campylobacter carried by flies and slugs acquired from local ruminant faeces.to assess whether flies and slugs acquire strains of campylobacter jejuni and campylobacter coli present in local ruminant faeces.201020337762
virulence and immunomodulatory roles of bacterial outer membrane vesicles.outer membrane (om) vesicles are ubiquitously produced by gram-negative bacteria during all stages of bacterial growth. om vesicles are naturally secreted by both pathogenic and nonpathogenic bacteria. strong experimental evidence exists to categorize om vesicle production as a type of gram-negative bacterial virulence factor. a growing body of data demonstrates an association of active virulence factors and toxins with vesicles, suggesting that they play a role in pathogenesis. one of the most ...201020197500
modification of the campylobacter jejuni flagellin glycan by the product of the cj1295 homopolymeric-tract-containing gene.the campylobacter jejuni flagellin protein is o-glycosylated with structural analogues of the nine-carbon sugar pseudaminic acid. the most common modifications in the c. jejuni 81-176 strain are the 5,7-di-n-acetylated derivative (pse5ac7ac) and an acetamidino-substituted version (pse5am7ac). other structures detected include o-acetylated and n-acetylglutamine-substituted derivatives (pse5am7ac8oac and pse5am7ac8glnnac, respectively). recently, a derivative of pseudaminic acid modified with a di ...201020338909
the internal transcribed spacer region, a new tool for use in species differentiation and delineation of systematic relationships within the campylobacter genus.the campylobacter genus consists of a number of important human and animal pathogens. although the 16s rrna gene has been used extensively for detection and identification of campylobacter species, there is currently limited information on the 23s rrna gene and the internal transcribed spacer (its) region that lies between the 16s and 23s rrna genes. we examined the potential of the 23s rrna gene and the its region to be used in species differentiation and delineation of systematic relationships ...201020348308
evidence for a lineage of virulent bacteriophages that target campylobacter.our understanding of the dynamics of genome stability versus gene flux within bacteriophage lineages is limited. recently, there has been a renewed interest in the use of bacteriophages as 'therapeutic' agents; a prerequisite for their use in such therapies is a thorough understanding of their genetic complement, genome stability and their ecology to avoid the dissemination or mobilisation of phage or bacterial virulence and toxin genes. campylobacter, a food-borne pathogen, is one of the organi ...201020353581
growth phase-dependent activation of the dccrs regulon of campylobacter jejuni.two-component systems are widespread prokaryotic signal transduction devices which allow the regulation of cellular functions in response to changing environmental conditions. the two-component system dccrs (cj1223c-cj1222c) of campylobacter jejuni is important for the colonization of chickens. here, we dissect the dccrs system in more detail and provide evidence that the sensor dccs selectively phosphorylates the cognate effector, dccr. microarray expression profiling, real-time reverse transcr ...201020348251
mutations in the rpsl gene are involved in streptomycin resistance in campylobacter coli.to characterize the mechanisms of streptomycin (str) resistance in campylobacter coli, we chose 17 isolates that were resistant to str, erythromycin (ery), or both, and the putative str resistance target genes rpsl, rrs, and gidb were analyzed for mutations. the presence of the aade gene encoding aminoglycoside 6-adenylyltransferase was also evaluated. to reveal putative connection between ery and str resistance mechanisms, 13 c. coli isolates initially susceptible to str and ery were exposed to ...201020370506
antimicrobial resistance, virulence, and genotypic profile comparison of campylobacter jejuni and campylobacter coli isolated from humans and retail meats.a total of 360 spatially and temporally related campylobacter isolates, including 168 from clinical human cases (campylobacter jejuni n = 148; campylobacter coli n = 20) and 192 from retail meats (c. jejuni n = 114; c. coli n = 78), were analyzed for antimicrobial susceptibilities, virulence, and genotypic profiles. ciprofloxacin-resistant c. jejuni was observed in 13.5% and 19% of the isolates from humans and retail chicken breasts, respectively. antimicrobial resistance to ciprofloxacin and er ...201020367499
prevalence and biomolecular characterization of campylobacter spp. isolated from retail meat.we estimated the prevalence of campylobacter spp. in retail meat (n = 352 samples; 104 chicken, 106 pork, and 142 beef) collected in campobasso, italy, comparing two microbiological methods. all the isolates were characterized by biomolecular techniques for epidemiological purposes. campylobacter isolation was performed by selective culture and membrane filtration methods. phenotypic and genotypic methods for genus and species identification were evaluated together with antimicrobial resistance ...201020377962
phenotypic and genotypic antimicrobial resistance profiles of campylobacter jejuni isolated from cattle, sheep, and free-range poultry faeces.minimum inhibitory concentrations (mic) of 13 antimicrobial agents were determined by broth microdilution for 72 campylobacter jejuni strains from livestock. twenty-three (31.9%) isolates were fully susceptible; all isolates were susceptible to erythromycin, chloramphenicol, streptomycin, gentamicin, sulfamethoxazole, and meropenem, and all but one to kanamycin. resistance to quinolones was highest (52.8%), reaching similar values among poultry, dairy cattle, and sheep, but lower in beef cattle. ...201020224816
phenotypic and genotypic antimicrobial resistance profiles of campylobacter jejuni isolated from cattle, sheep, and free-range poultry faeces.minimum inhibitory concentrations (mic) of 13 antimicrobial agents were determined by broth microdilution for 72 campylobacter jejuni strains from livestock. twenty-three (31.9%) isolates were fully susceptible; all isolates were susceptible to erythromycin, chloramphenicol, streptomycin, gentamicin, sulfamethoxazole, and meropenem, and all but one to kanamycin. resistance to quinolones was highest (52.8%), reaching similar values among poultry, dairy cattle, and sheep, but lower in beef cattle. ...201020224816
fluorescence in situ hybridization method using a peptide nucleic acid probe for identification of salmonella spp. in a broad spectrum of samples.a fluorescence in situ hybridization (fish) method for the rapid detection of salmonella spp. using a novel peptide nucleic acid (pna) probe was developed. the probe theoretical specificity and sensitivity were both 100%. the pna-fish method was optimized, and laboratory testing on representative strains from the salmonella genus subspecies and several related bacterial species confirmed the predicted theoretical values of specificity and sensitivity. the pna-fish method has been successfully ad ...201020453122
prevalence and antimicrobial resistance of campylobacter isolates from humans and chickens in bahir dar, ethiopia.in this study, the isolation and antimicrobial resistance of campylobacter jejuni and campylobacter coli strains from chickens and humans in bahir dar, ethiopia, were analyzed. two hundred and ten human and 220 chicken samples were analyzed between october 2007 and april 2008. seventeen human and 160 chicken campylobacter species were isolated. the overall prevalence of thermophilic campylobacters was 8% and 72.7% in humans and chickens, respectively. in humans, 94.1% of the isolates were c. jej ...201020482228
occurrence of thermotolerant campylobacter species in surface waters of a mediterranean area and in its prevailing pollution sources.to determine the prevalence of campylobacter in surface waters of a highly populated mediterranean area.201020408911
repeated therapeutic dosing selects macrolide-resistant campylobacter spp. in a turkey facility.this study assessed the effects of the therapeutic use of tylan® in a large-scale turkey production facility on the selection of macrolide-resistant campylobacter.201020497488
fluoroquinolone resistance detection in campylobacter coli and campylobacter jejuni by luminex xmap technology.the proportion of campylobacter spp. isolates that are resistant to fluoroquinolones, the drugs of choice for campylobacteriosis, has been increasing worldwide. we developed an innovative method based on a luminex xmap dna suspension array that allows the identification of campylobacter species and, simultaneously, the detection of the most common point mutation in the gyra gene (substitution from threonine 86 to isoleucine 86) that is responsible for fluoroquinolone resistance. ninety-six campy ...201020500084
prevalence and antimicrobial resistance of campylobacter jejuni and campylobacter coli isolated from cattle between 2002 and 2006 in france.feces from 2,255 cattle (calves, young beef cattle, and culled cows) were collected at slaughter from nine departments across france. campylobacter was recovered from 16.5% of the 2,255 samples (c. jejuni from 12.8% and c. coli from 3.7%), predominantly from calves. antimicrobial resistance to six antibiotics of medical and/or veterinary interest was tested with the e-test. resistance to tetracycline was found in most isolates (52.8% of c. jejuni isolates and 88.1% of c. coli isolates) in contra ...201020501032
investigation of water washes suitable for very small meat plants to reduce pathogens on beef surfaces.water washing with a handheld hose was performed on beef surfaces to ascertain the most effective combination of methods needed to remove potentially harmful microorganisms. for these experiments, beef brisket surfaces were experimentally inoculated with a fecal slurry containing escherichia coli o157:h7, salmonella typhimurium, campylobacter coli, and campylobacter jejuni. in a pilot study, surfaces were washed with cold water (15 degrees c) at various water pressures, spray distances, applicat ...201020501042
detection of campylobacter jejuni by culture and real-time pcr in a french cohort of patients with guillain-barre syndrome.bacteriological culture and real-time pcr (rt-pcr) were used to detect campylobacter jejuni in fecal samples from a french cohort of 237 patients with guillain-barré syndrome (gbs). we provide evidence that diverse serotypes and genotypes of c. jejuni are a major trigger of gbs in france.201020410350
three-hour molecular detection of campylobacter, salmonella, yersinia, and shigella species in feces with accuracy as high as that of culture.campylobacter jejuni and salmonella, shigella, and yersinia species (along with shiga toxin-producing escherichia coli) are the most common causes of acute bacterial diarrheal disease in the united states. current detection techniques are time-consuming, limiting usefulness for patient care. we developed and validated a panel of rapid pcr assays for the detection and identification of c. jejuni, c. coli, salmonella, and yersinia species and shigella and enteroinvasive e. coli in stool samples. a ...201020519461
host association of campylobacter genotypes transcends geographic variation.genetic attribution of bacterial genotypes has become a major tool in the investigation of the epidemiology of campylobacteriosis and has implicated retail chicken meat as the major source of human infection in several countries. to investigate the robustness of this approach to the provenance of the reference data sets used, a collection of 742 campylobacter jejuni and 261 campylobacter coli isolates obtained from united kingdom-sourced chicken meat was established and typed by multilocus seque ...201020525862
molecular typing of campylobacter jejuni and c. coli from chickens and patients with gastritis or guillain-barré syndrome based on multilocus sequence types and pulsed-field gel electrophoresis patterns.campylobacter jejuni has recently been noted as the most common cause of bacterial foodborne diseases in japan. in the present study, we determined st types of c. jejuni and campylobacter coli isolated from chickens and patients with enteritis or gbs in japan and thailand. c. jejuni from chickens, enteritis, and gbs exhibited divergent st types and included several novel types in addition to worldwide common types. c. coli from enteritis was also divergent. novel st types may represent unidentif ...201020536735
comparison of real-time pcr assays for detection, quantification, and differentiation of campylobacter jejuni and campylobacter coli in broiler neck skin samples.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 betwe ...201020537260
fluoroquinolone resistance in campylobacter.campylobacter is a commensal in poultry, and therefore, poultry and poultry products are major sources of campylobacter infections in humans. fluoroquinolones inhibit the growth of campylobacter and other microorganisms by binding to bacterial dna gyrase and dna topoisomerase iv. these enzymes are associated with bacterial transcription, replication, and chromosome condensation and segregation. selection pressure in the presence of fluoroquinolones rapidly leads to resistance in campylobacter, d ...201020537275
campylobacter jejuni and campylobacter coli in children from communities in northeastern brazil: molecular detection and relation to nutritional status.this study determined the prevalence of campylobacter jejuni/coli and its relation with nutritional status in children from northeastern brazil. this was a case-control study design. stool samples were evaluated for hipo (c. jejuni), ask (c. coli), and cdtabc (c. jejuni's cytolethal distending toxin) genes. the nutritional status from these children was assessed by anthropometric measures and z-scores. c. jejuni and c. coli were detected in 9.6% (8/83) and 6.0% (5/83) in the diarrhea group and i ...201020542202
performance of the oxoid m.i.c.evaluator strips compared with the etest assay and bsac agar dilution.the oxoid m.i.c.evaluator (m.i.c.e; thermo fisher scientific) comprises an antibiotic gradient on a plastic support. we compared its performance with etest--a similar product--using bsac agar dilution as a reference.201020562429
prevalence and antimicrobial resistance of campylobacter species isolated from chicken carcasses during processing in iran.the objective of this study was to determine the prevalence and antimicrobial resistance of campylobacter spp. isolated from chicken carcasses during different stages of broiler processing in a major commercial poultry processing plant in southwestern iran. overall, 84 chicken carcasses were sampled from 4 sites along the processing line during a total of 7 visits. in addition, 14 water samples from the chiller tank were taken. using the cultural method, 186 of 336 (55.4%) carcasses were positiv ...201020371855
identification of campylobacter jejuni genes involved in its interaction with epithelial cells.campylobacter jejuni is the leading cause of infectious gastroenteritis in industrialized nations. its ability to enter and survive within nonphagocytic cells is thought to be very important for pathogenesis. however, little is known about the c. jejuni determinants that mediate these processes. through an extensive transposon mutagenesis screen, we have identified several loci that are required for c. jejuni efficient entry and survival within epithelial cells. among these loci, insertional mut ...201020515930
identification and characterization of a new ferric enterobactin receptor, cfrb, in campylobacter.the ferric enterobactin (feent) receptor cfra is present in the majority of campylobacter jejuni isolates and is responsible for high-affinity iron acquisition. our recent work and that of others strongly suggested the existence of another feent uptake system in campylobacter. here we have identified and characterized a new feent receptor (designated cfrb) using both in vitro and in vivo systems. cfrb, a homolog of c. jejuni nctc 11168 cj0444, shares approximately 34% of amino acid identity with ...201020585060
campylobacter spp among children with acute diarrhea attending mulago hospital in kampala--uganda.campylobacter infections occur worldwide. a recent study in kampala, uganda, found that 87% of broiler chickens had campylobacter jejuni; these are potential source of human infection. 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.200920589152
prevalence and antimicrobial resistance in campylobacter spp. isolated from retail chicken in two health units in ontario.campylobacter is an important enteric pathogen of humans and can cause diarrhea, fever, and abdominal pain. campylobacter infections have frequently been associated with the handling and consumption of raw and undercooked poultry. antimicrobial resistance among campylobacter strains is of concern in the treatment of campylobacteriosis in vulnerable populations. a 2-year multidisciplinary study was conducted in the perth and wellington-dufferin-guelph public health units in ontario, canada, to in ...201020615345
carry-over of thermophilic campylobacter spp. between sequential and adjacent poultry flocks.nineteen flocks of four poultry species were monitored at a veterinary field station to investigate the distribution and spread of campylobacter genotypes between sequential and adjacent flocks. caecal and liver samples were obtained at frequent intervals from birds of all flocks and examined for campylobacter. amplified fragment length polymorphism (aflp) analysis was performed to genotype campylobacter isolates. of the 1643 caecal and liver samples investigated, 452 (27.5%) caecal samples and ...201120619560
screening for several potential pathogens in feral pigeons (columba livia) in madrid.pathogens with the zoonotic potential to infect humans, such as campylobacter jejuni, campylobacter coli and chlamydophila psittaci, can be found in feral pigeons (columba livia). given the high density of these birds in the public parks and gardens of most cities, they may pose a direct threat to public health.201020569487
isolation and characterization of campylobacter spp. from antarctic fur seals (arctocephalus gazella) at deception island, antarctica.the presence of campylobacter spp. was investigated in 41 antarctic fur seals (arctocephalus gazella) and 9 weddell seals (leptonychotes weddellii) at deception island, antarctica. infections were encountered in six antarctic fur seals. the isolates, the first reported from marine mammals in the antarctic region, were identified as campylobacter insulaenigrae and campylobacter lari.201020639356
survival of campylobacter jejuni under conditions of atmospheric oxygen tension with the support of pseudomonas spp.campylobacter jejuni is a major food-borne pathogen. despite causing enteritis in humans, it is a well-adapted intestinal microorganism in animals, hardly ever generating disease symptoms. nevertheless, as a true microaerophilic microorganism it is still puzzling how campylobacter cells can survive on chicken meat, the main source of human infection. in this study, we demonstrate that c. jejuni is able to withstand conditions of atmospheric oxygen tension when cocultured with pseudomonas species ...201020639377
genotyping and pcr detection of potential virulence genes in campylobacter jejuni and campylobacter coli isolates from different sources in poland.the prevalence of potential virulence markers was determined among the population of polish campylobacter jejuni and campylobacter coli isolates from children, chickens, pigs and dogs. the presence of the flaa, flab, cdta, cdtb, cdtc, cdtabc, virb11, and cj0588 genes among 74 c. jejuni and 15 c. coli isolates was detected by pcr. high prevalence of five different putative virulence and toxin genes (flaa, cdta, cdtb, cdtc, and cj0588) was found among isolates obtained from children, chickens and ...201020490760
characterization of n-linked protein glycosylation in helicobacter pullorum.the first bacterial n-linked glycosylation system was discovered in campylobacter jejuni, and the key enzyme involved in the coupling of glycan to asparagine residues within the acceptor sequon of the glycoprotein is the oligosaccharyltransferase pglb. emerging genome sequence data have revealed that pglb orthologues are present in a subset of species from the deltaproteobacteria and epsilonproteobacteria, including three helicobacter species: h. pullorum, h. canadensis, and h. winghamensis. in ...201020581208
simple and rapid detection of campylobacter spp. in naturally contaminated chicken-meat samples by combination of a two-step enrichment method with an immunochromatographic assay.a simple and rapid method to detect campylobacter spp. in chicken-meat samples was established. this method consisted of a combination of a two-step enrichment method with a commercially available immunochromatographic assay, named nh immunochromato campylobacter (nh ic campy, nippon meat packers, ibaraki, japan), which is able to detect campylobacter antigen in an enrichment culture within 15 min. the enrichment method did not require much blood or a particular system of generating a microaerob ...201020621373
antimicrobial resistance in campylobacter coli selected by tylosin treatment at a pig farm.limited knowledge is available regarding the dynamics of macrolide resistance under farm conditions with natural campylobacter populations. we examined the dynamics of antimicrobial resistance in campylobacter coli at a large pig farm. faeces were sampled from untreated sows and piglets (n=57), weaned pigs treated with tylosin (n=68) and pigs of the same group 3-5 weeks after withdrawal of tylosin (n=15). additionally, 48 weaned pigs were sampled after tylosin had not been administered for 7 mon ...201020684867
evolutionary dynamics of complete campylobacter pan-genomes and the bacterial species concept.defining bacterial species and understanding the relative cohesiveness of different components of their genomes remains a fundamental problem in microbiology. bacterial species tend to be comprised of both a set of core and dispensable genes, with the sum of these two components forming the species pan-genome. the role of the core and dispensable genes in defining bacterial species and the question of whether pan-genomes are finite or infinite remain unclear. here we demonstrate, through the ana ...201020688752
genotype and antibiotic resistance analyses of campylobacter isolates from ceca and carcasses of slaughtered broiler flocks.to obtain genetic information about campylobacter jejuni and campylobacter coli from broilers and carcasses at slaughterhouses, we analyzed and compared 340 isolates that were collected in 2008 from the cecum right after slaughter or from the neck skin after processing. we performed rpob sequence-based identification, multilocus sequence typing (mlst), and flab sequence-based typing; we additionally analyzed mutations within the 23s rrna and gyra genes that confer resistance to macrolide and qui ...201020709846
occurrences of thermophilic campylobacter in pigs slaughtered at morogoro slaughter slabs, tanzania.occurrences of thermophlic campylobacter in pigs and pig carcasses was investigated in a cross-sectional study that was carried out in three selected slaughter slabs in morogoro municipality, tanzania. before sampling, the slab hygiene, slaughter, carcass dressing, and meat handling was assessed. fecal samples were collected from 66 slaughter pigs at the kill floor. after slaughter, a 100-cm(2) area on medial surface of the thigh muscles of dressed carcasses was sampled using sterile cotton swab ...201120730490
characterization of erythromycin-resistant porcine isolates of campylobacter coli.erythromycin-resistant campylobacter organisms were isolated from swine, and their resistance to the antibiotic was characterized. one hundred fourteen campylobacter organisms were isolated from 572 swine intestinal samples. all isolates were identified as campylobacter coli by sequence analysis of 16s rrna gene and polymerase chain reactions with primers specific to hippuricase gene in campylobacter jejuni and aspartokinase gene in c. coli. minimal inhibitory concentrations (mics) of erythromyc ...201020735175
study of the molecular mechanisms involved in high-level macrolide resistance of spanish campylobacter jejuni and campylobacter coli strains.to investigate the molecular mechanisms involved in the high-level erythromycin resistance of clinical spanish campylobacter jejuni and campylobacter coli strains.201020647243
implications of faecal indicator bacteria for the microbiological assessment of roof-harvested rainwater quality in southeast queensland, australia.the study aimed to evaluate the suitability of escherichia coli, enterococci, and clostridium perfringens for assessing the microbiological quality of roof-harvested rainwater and assessing whether the concentrations of these faecal indicators can be used to predict the presence or absence of specific zoonotic bacterial or protozoan pathogens. from a total of 100 samples tested, 58%, 83%, and 46% of samples were found to be positive for, respectively, e. coli, enterococci, and clostridium perfri ...201020657617
the in vivo efficacy of two administration routes of a phage cocktail to reduce numbers of campylobacter coli and campylobacter jejuni in chickens.poultry meat is one of the most important sources of human campylobacteriosis, an acute bacterial enteritis which is a major problem worldwide. campylobacter coli and campylobacter jejuni are the most common campylobacter species associated with this disease. these pathogens live in the intestinal tract of most avian species and under commercial conditions they spread rapidly to infect a high proportion of the flock, which makes their treatment and prevention very difficult. bacteriophages (phag ...201020809975
accuracy of the api campy system, the vitek 2 neisseria-haemophilus (nh) card and the matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (maldi-tof ms) for the identification of campylobacter and related organisms.abstract biochemical identification of campylobacter and related organisms is not always specific and may lead to diagnostic errors. the api campy, the vitek 2 system and matrix-assisted desorption/ionisation time-of-flight mass spectrometry (maldi-tof ms) are commercially available methods that are routinely used for the identification of these microorganisms. in the present study, we used 224 clinical isolates and 10 reference strains previously identified by multiple pcr assays, whole-cell pr ...201020673261
obstacles of multiplex real-time pcr for bacterial 16s rdna: primer specifity and dna decontamination of taq polymerase.background: the detection of a broad range of bacteria by pcr is applied for the screening of blood and blood products with special attention to platelet concentrates. for practical use it is desirable that detection systems include gram-positive, gram-negative and non-gram-stainable bacteria. it is quite challenging to achieve high sensitivity along with a clear negative control with pcr reagents, because especially taq polymerase is contaminated with traces of bacterial dna. methods: bacterial ...201020737013
comparison of premier campy enzyme immunoassay (eia), prospect campylobacter eia, and immunocard stat! campy tests with culture for laboratory diagnosis of campylobacter enteric infections.campylobacter enteritis is a food-borne or waterborne illness caused almost exclusively by campylobacter jejuni and, to a lesser extent, by campylobacter coli. these organisms produce indistinguishable clinical diseases and together represent the second most common cause of bacterial diarrhea in the united states and the leading cause of enteric infection throughout the world. the conventional approach to the laboratory diagnosis of campylobacter enteritis is based on the recovery of the organis ...201020810765
prevalence, antibiograms, and transferable tet(o) plasmid of campylobacter jejuni and campylobacter coli isolated from raw chicken, pork, and human clinical cases in korea.the antibiotic resistance patterns and prevalence of the transferable tet(o) plasmid were investigated in campylobacter jejuni and campylobacter coli isolates from raw chicken, pork, and humans with clinical campylobacteriosis. a total of 180 c. jejuni and c. coli isolates were identified, and the prevalence rates of c. jejuni and c. coli in raw chicken samples were 83% (83 of 100) and 73% (73 of 100), respectively. twelve percent (6 of 50) and 10% (5 of 50) of pork samples were contaminated wit ...201020819352
survival of campylobacter jejuni and campylobacter coli on retail broiler meat stored at -20, 4, or 12 degrees c and development of weibull models for survival.survival of campylobacter jejuni and campylobacter coli isolated from broiler meat was investigated and modeled on retail breast meat. meat portions were inoculated with c. jejuni or c. coli at 6.4 to 6.8 log cfu/g followed by storage at -20 degrees c for 84 days or at 4 or 12 degrees c for 14 days. kinetic data within a species and temperature were fitted to the weibull model. when >or=70% of the residuals were in an acceptable prediction zone from -1 (fail-safe) to 0.5 (fail-dangerous) log uni ...201020819353
differences in methylation at gatc sites in genomic dna of campylobacter coli from turkeys and swine.a significant fraction (46/108, 43%) of swine isolates of campylobacter coli but none of 81 isolates of c. coli from turkeys had genomic dna that was resistant to digestion by mboi, suggesting methylation of adenines at gatc sites. no consistent association was noted between antimicrobial resistance and mboi resistance. seven swine-associated multilocus sequence typing-based sequence types (sts) were detected among multiple isolates with mboi-resistant dna. the data suggest host-associated dna m ...201020833787
ribosomal operon intergenic sequence region (isr) heterogeneity in campylobacter coli and campylobacter jejuni.the intergenic sequence regions (isr) between the 16s and 23s genes of campylobacter jejuni and campylobacter coli are markedly different for each species. however, in the genomic sequence for camp. coli rm2228, two rrna operons have an isr that is characteristic of camp. coli, and the third operon is characteristic of camp. jejuni. the aim of this study was to determine the prevalence of isr heterogeneity in these organisms.201020849393
direction of neutrophil movements by campylobacter-infected intestinal epithelium.campylobacter jejuni and campylobacter coli together represent the leading bacterial cause of human enteritis. however, the pathogenesis of this disease is poorly understood. infection results in the formation of crypt abscesses resulting from the migration of neutrophils across the intestinal epithelium and into the intestinal crypts. in this study, we model this process in vitro and show that campylobacter infection of epithelium results in a quantifiable increase in the directed movement of n ...201020934530
fragmentation of the large subunit ribosomal rna gene in oyster mitochondrial genomes.discontinuous genes have been observed in bacteria, archaea, and eukaryotic nuclei, mitochondria and chloroplasts. gene discontinuity occurs in multiple forms: the two most frequent forms result from introns that are spliced out of the rna and the resulting exons are spliced together to form a single transcript, and fragmented gene transcripts that are not covalently attached post-transcriptionally. within the past few years, fragmented ribosomal rna (rrna) genes have been discovered in bilatera ...201020813041
prevalence of campylobacter jejuni, campylobacter coli and enteric helicobacter in domestic and free living birds in north-western italy.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 carr ...201020814861
the glycerophospho metabolome and its influence on amino acid homeostasis revealed by brain metabolomics of gde1(-/-) mice.gde1 is a mammalian glycerophosphodiesterase (gde) implicated by in vitro studies in the regulation of glycerophophoinositol (gropins) and possibly other glycerophospho (grop) metabolites. here, we show using untargeted metabolomics that gropins is profoundly (>20-fold) elevated in brain tissue from gde1(-/-) mice. furthermore, two additional grop metabolites not previously identified in eukaryotic cells, glycerophosphoserine (gropser) and glycerophosphoglycerate (gropgate), were also highly ele ...201020797612
antimicrobial resistance of campylobacter isolates from retail meat in the united states between 2002 and 2007.the emergence of antimicrobial resistance in campylobacter spp. has been a growing public health concern globally. the objectives of this study were to determine the prevalence, antimicrobial susceptibility, and genetic relatedness of campylobacter spp. recovered by the national antimicrobial resistance monitoring system (narms) retail meat program. retail meat samples (n = 24,566) from 10 u.s. states collected between 2002 and 2007, consisting of 6,138 chicken breast, 6,109 ground turkey, 6,171 ...201020971875
evolution and population structure of salmonella enterica serovar newport.salmonellosis caused by salmonella enterica serovar newport is a major global public health concern, particularly because s. newport isolates that are resistant to multiple drugs (mdr), including third-generation cephalosporins (mdr-ampc phenotype), have been commonly isolated from food animals. we analyzed 384 s. newport isolates from various sources by a multilocus sequence typing (mlst) scheme to study the evolution and population structure of the serovar. these were compared to the populatio ...201020935094
detection of gyra mutation among clinical isolates of campylobacter jejuni isolated in egypt by mama-pcr.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 posi ...201021045366
prevalence and characterization of campylobacter spp. isolated from domestic and imported poultry meat in korea, 2004–2008.campylobacteriosis in humans is primarily caused by handling or consuming contaminated poultry or their products. the aims of this study were to estimate the prevalence of campylobacter spp. in domestic and imported poultry meat in korea and to further characterize the obtained isolates. from 2004 to 2008, a total of 475 domestic and 867 imported raw poultry meat samples were examined for the presence of campylobacter spp. among 475 domestic poultry meat samples, campylobacter jejuni and campylo ...201020939742
temperature affects sole carbon utilization patterns of campylobacter coli 49941.campylobacter spp. are small, asaccharolytic bacteria exhibiting unique nutritional and environmental requirements. campylobacter spp. exist as commensal organisms in some animal species, yet are estimated to be the most common causative agents of foodborne illness in humans. c. jejuni is most often associated with poultry, while c. coli are more frequently associated with swine. temperature has been suggested to trigger potential colonization or virulence factors in c. jejuni, and recent studie ...201020981547
molecular characterization and environmental mapping of campylobacter isolates in a subset of intensive poultry flocks in ireland.irish breeder and intensive broiler flocks together with the corresponding poultry farm environment were sampled for the presence of campylobacter with the aim of identifying potential sources and transmission routes of poultry flock contamination. the genetic diversity of a subset of campylobacter isolates was examined by analysis of the flaa-short variable region (svr). additional discrimination for a further subset of these isolates was achieved using multilocus sequence typing (mlst) analysi ...201021039133
microbiological quality of blue mussels (mytilus edulis) in nunavik, quebec: a pilot study.this pilot study was aimed at documenting the presence of fecal indicators and enteric pathogens in blue mussels (mytilus edulis) from 6 communities in nunavik, quebec. one to four 2 kg samples of mussels were collected at low tide in each community. samples were investigated by enumeration methods for the fecal indicators enterococci, escherichia coli, f-specific coliphages, clostridium perfringens, and by molecular identification for the pathogens norovirus, salmonella spp., campylobacter jeju ...201021076488
prevalence, numbers and antimicrobial susceptibilities of salmonella serovars and campylobacter spp. in retail poultry in phnom penh, cambodia.salmonella and campylobacter are common bacterial pathogens associated with human gastro-enteritis; and raw poultry is considered to be an important source of these bacteria. to evaluate whether the salmonella serovars and campylobacter spp. bacteria could be monitored for the purpose of microbial presence, enumeration and antimicrobial resistance in raw poultry, 152 poultry carcasses were randomly selected from 10 markets in retail outlets of phnom penh during march 2006 to february 2007. the m ...201021060246
campylobacter genotypes from poultry transportation crates indicate a source of contamination and transmission.crates used to transport live poultry can be contaminated with campylobacter, despite periodic sanitization, and are potential vectors for transmission between flocks. we investigated the microbial contamination of standard and silver ion containing crates in normal use and the genetic structure of associated campylobacter populations.201021059158
campylobacter genotypes from poultry transportation crates indicate a source of contamination and transmission.crates used to transport live poultry can be contaminated with campylobacter, despite periodic sanitization, and are potential vectors for transmission between flocks. we investigated the microbial contamination of standard and silver ion containing crates in normal use and the genetic structure of associated campylobacter populations.201021059158
phenotypic and genotypic heterogeneity of campylobacter coli within individual pigs at farm and slaughter in the us.the aim of this study was to determine the phenotypic and genotypic diversity of multiple campylobacter isolates (n = 3 per sample) present within individual (heterogeneity) pig faecal and carcass samples at farm and slaughter, respectively. we isolated 1459 campylobacter coli (1110 on farm and 349 from slaughter) from 908 pigs and 757 carcasses and characterized them for their antimicrobial susceptibility profile to a panel of six antimicrobials using the agar dilution method. overall, we detec ...201021083823
interbacterial macromolecular transfer by the campylobacter fetus subsp. venerealis type iv secretion system.we report here the first demonstration of intra- and interspecies conjugative plasmid dna transfer for campylobacter fetus. gene regions carried by a campylobacter coli plasmid were identified that are sufficient for conjugative mobilization to escherichia coli and c. fetus recipients. a broader functional range is predicted. efficient dna transfer involves the virb9 and vird4 genes of the type iv bacterial secretion system encoded by a pathogenicity island of c. fetus subsp. venerealis. complem ...201021115658
typing of campylobacter jejuni and campylobacter coli isolated from live broilers and retail broiler meat by flaa-rflp, mlst, pfge and rep-pcr.we analyzed 100 campylobacter spp. isolates (c. jejuni and c. coli) from grenada, puerto rico and alabama, which were collected from live broilers or retail broiler meat. we analyzed these isolates with four molecular typing methods: restriction fragment length polymorphism of the flaa gene (flaa-rflp), multilocus sequence typing (mlst), pulsed-field gel electrophoresis (pfge), and automated repetitive extragenic palindromic polymerase chain reaction (rep-pcr) using the diversilab system. all me ...201021130125
independent evolution of the core and accessory gene sets in the genus neisseria: insights gained from the genome of neisseria lactamica isolate 020-06.the genus neisseria contains two important yet very different pathogens, n. meningitidis and n. gonorrhoeae, in addition to non-pathogenic species, of which n. lactamica is the best characterized. genomic comparisons of these three bacteria will provide insights into the mechanisms and evolution of pathogenesis in this group of organisms, which are applicable to understanding these processes more generally.201021092259
prevalence and antimicrobial resistance profile of campylobacter spp. isolated from conventional and antimicrobial-free swine production systems from different u.s. regions.we conducted a study to compare the prevalence and antimicrobial resistance profile of campylobacter isolated from 34 farm-slaughter pair cohorts of pigs raised in conventional and antimicrobial-free (abf) production systems. isolates originated from four different states of two geographic regions (region 1--ohio and michigan; region 2--wisconsin and iowa). a total of 838 fecal and 1173 carcass samples were examined. campylobacter isolates were speciated using multiplex polymerase chain reaction ...201021133777
in vitro activity and single-step mutational analysis of rifamycin sv tested against enteropathogens associated with traveler's diarrhea and clostridium difficile.rifamycin sv is a broad-spectrum, poorly absorbed antimicrobial agent that, when coupled with mmx technology, is being targeted for the oral treatment of traveler's diarrhea (td) and clostridium difficile-associated disease (cdad). rifamycin sv was tested for activity against 911 td-associated enteropathogens and 30 c. difficile isolates collected from several global surveillance studies. rifamycin sv demonstrated similar antimicrobial activity levels against the enterobacteriaceae, with mic₅₀ v ...201021149623
in vitro activity and single-step mutational analysis of rifamycin sv tested against enteropathogens associated with traveler's diarrhea and clostridium difficile.rifamycin sv is a broad-spectrum, poorly absorbed antimicrobial agent that, when coupled with mmx technology, is being targeted for the oral treatment of traveler's diarrhea (td) and clostridium difficile-associated disease (cdad). rifamycin sv was tested for activity against 911 td-associated enteropathogens and 30 c. difficile isolates collected from several global surveillance studies. rifamycin sv demonstrated similar antimicrobial activity levels against the enterobacteriaceae, with mic₅₀ v ...201021149623
prevalence and risk factors associated with campylobacter infections in broiler flocks in shiraz, southern iran.campylobacter species are among the most common bacterial causes of human gastroenteritis in many countries, and poultry meat is considered as a major source of human campylobacteriosis. the present study was conducted to determine the prevalence of infection by campylobacter jejuni and campylobacter coli in broiler flocks in shiraz and to investigate the possible risk factors for the campylobacter infections in this area. for detection of campylobacter, multiplex polymerase chain reaction (mpcr ...201021131089
enteric campylobacteria and rna viruses associated with healthy and diarrheic humans in the chinook health region of southwestern alberta, canada.the presence of campylobacter species and enteric rna viruses in stools from diarrheic (n = 442) and healthy (n = 58) humans living in southwestern alberta was examined (may to october 2005). a large number of diarrheic individuals who were culture negative for c. jejuni (n = 54) or c. coli (n = 19) were pcr positive for these taxa. overall detection rates for c. jejuni and c. coli in diarrheic stools were 29% and 5%, respectively. in contrast, 3% and 0% of stools from healthy humans were positi ...201021106791
enteric campylobacteria and rna viruses associated with healthy and diarrheic humans in the chinook health region of southwestern alberta, canada.the presence of campylobacter species and enteric rna viruses in stools from diarrheic (n = 442) and healthy (n = 58) humans living in southwestern alberta was examined (may to october 2005). a large number of diarrheic individuals who were culture negative for c. jejuni (n = 54) or c. coli (n = 19) were pcr positive for these taxa. overall detection rates for c. jejuni and c. coli in diarrheic stools were 29% and 5%, respectively. in contrast, 3% and 0% of stools from healthy humans were positi ...201021106791
evaluation of salmonella-vectored campylobacter peptide epitopes for reduction of campylobacter jejuni in broiler chickens.campylobacter is a leading cause of bacterial gastroenteritis in humans and is often linked to contaminated poultry products. live salmonella vectors expressing three linear peptide epitopes from campylobacter proteins cj0113 (omp18/cjad), cj0982c (cjaa), and cj0420 (ace393) were administered to chicks by oral gavage on the day of hatch, and the chicks were challenged with campylobacter jejuni on day 21. all three candidate vaccines produced consistent humoral immune responses with high levels o ...201021177910
evaluation of salmonella-vectored campylobacter peptide epitopes for reduction of campylobacter jejuni in broiler chickens.campylobacter is a leading cause of bacterial gastroenteritis in humans and is often linked to contaminated poultry products. live salmonella vectors expressing three linear peptide epitopes from campylobacter proteins cj0113 (omp18/cjad), cj0982c (cjaa), and cj0420 (ace393) were administered to chicks by oral gavage on the day of hatch, and the chicks were challenged with campylobacter jejuni on day 21. all three candidate vaccines produced consistent humoral immune responses with high levels o ...201021177910
novel glycosylation sites localized in campylobacter jejuni flagellin flaa by liquid chromatography electron capture dissociation tandem mass spectrometry.glycosylation of flagellin in campylobacter jejuni is essential for motility and virulence. it is well-known that flagellin from c. jejuni 81-176 is glycosylated by pseudaminic acid and its acetamidino derivative, and that campylobactor coli vc167 flagellin is glycosylated by legionaminic acid and its derivatives. recently, it was shown, by use of a metabolomics approach, that c. jejuni 11168 is glycosylated by dimethyl glyceric acid derivatives of pseudaminic acid, but the sites of glycosylatio ...201121158479
drinking water as the source of campylobacter coli infection in grandparent heavy breeders.the aim of the present study was the molecular identification of a common source of infection of campylobacter coli in two grandparent breeder farms. campylobacter jejuni and c. coli were isolated from well water and cloacal swabs from grandparent chickens. colonies were genotyped using restriction fragment length polymorphism-flaa gene, pulsed field gel electrophoresis and multi-locus sequence typing. the same genotype of c. coli was found in both farms and in the well from which drinking water ...201021154058
new methods for detection of campylobacters in stool samples in comparison to culture.campylobacter species, especially campylobacter jejuni and campylobacter coli, are a major cause of human bacterial enteritis. current detection in stools is done essentially by culture on selective and nonselective media with filtration. these methods were compared to 2 molecular biology methods, an in-house real-time pcr and a multiplex pcr named seeplex diarrhea ace detection, and 3 immunoenzymatic methods, premier campy, ridascreen campylobacter, and immunocard stat!campy. out of 242 stool s ...201121209172
novel glycosylation sites localized in campylobacter jejuni flagellin flaa by liquid chromatography electron capture dissociation tandem mass spectrometry.glycosylation of flagellin in campylobacter jejuni is essential for motility and virulence. it is well-known that flagellin from c. jejuni 81-176 is glycosylated by pseudaminic acid and its acetamidino derivative, and that campylobactor coli vc167 flagellin is glycosylated by legionaminic acid and its derivatives. recently, it was shown, by use of a metabolomics approach, that c. jejuni 11168 is glycosylated by dimethyl glyceric acid derivatives of pseudaminic acid, but the sites of glycosylatio ...201121158479
description and burden of travel-related cases caused by enteropathogens reported in a canadian community.risk of infections by enteropathogens among individuals traveling outside their country of residence is considered important. such travel-related cases (trc) have been poorly estimated and described in canada.201021199137
evolution of an agriculture-associated disease causing campylobacter coli clade: evidence from national surveillance data in scotland.the common zoonotic pathogen campylobacter coli is an important cause of bacterial gastroenteritis worldwide but its evolution is incompletely understood. using multilocus sequence type (mlst) data of 7 housekeeping genes from a national survey of campylobacter in scotland (2005/6), and a combined population genetic-phylogenetics approach, we investigated the evolutionary history of c. coli. genealogical reconstruction of isolates from clinical infection, farm animals and the environment, reveal ...201021179537
detection, isolation, and characterization of helicobacter species from the gastrointestinal tract of the brushtail possum.the presence of helicobacter species in australian marsupials was examined systematically using microscopy, culture, and pcr in different regions of the gastrointestinal tract (git) and in the liver of brushtail possums (btps) (trichosurus vulpecula), a common australian marsupial that feeds on eucalyptus leaves. the spatial distribution of helicobacter species in the git sections also was examined microscopically in silver-stained sections and by fluorescent in situ hybridization (fish) using a ...201121216901
arcobacter population dynamics in pigs on farrow-to-finish farms.healthy pigs are an important reservoir for the emerging human pathogen arcobacter which can result in contamination of porcine carcasses and pork and the spread of arcobacters into the environment. up to now, the excretion of arcobacters by pigs has been studied, but information about the transmission routes in fattening pigs is lacking. the present study aimed to elucidate the arcobacter population dynamics in pigs during the fattening period on four farrow-to-finish farms. on each farm, 30 cl ...201121216902
in vivo recognition of the feca3 target promoter by helicobacter pylori nikr.in helicobacter pylori, the transcriptional regulator hpnikr represses transcription of the feca3 gene by binding to two adjacent operators spanning a region of almost 80 nucleotides along the feca3 promoter in a nickel-dependent manner. by employing hydroxyl radical footprinting, we mapped the protected nucleotides within each operator. three short sequences rich in a and t nucleotides were identified within each operator, comprising just 24 bases for both operators, with 4 or 5 protected bases ...201121216998
adherence, invasion, toxigenic, and chemotactic properties of mexican campylobacter strains.to determine the virulence factors of mexican wild-type strains of campylobacter jejuni and campylobacter coli, 31 wild-type strains were isolated from food and from humans. the production of cytolethal distending toxin and the adherence and invasion capabilities of these strains were assayed in vero cells. hard agar plugs with repellents and attractants were used to examine chemotaxis. mueller-hinton agar with supplements was used for motility analysis and to measure hemolytic activity. nine st ...201021219724
introgression in the genus campylobacter: generation and spread of mosaic alleles.horizontal genetic exchange strongly influences the evolution of many bacteria, substantially contributing to difficulties in defining their position in taxonomic groups. in particular, how clusters of related bacterial genotypes - currently classified as microbiological species - evolve and are maintained remains controversial. the nature and magnitude of gene exchange between two closely related (approx. 15 % nucleotide divergence) microbiologically defined species, campylobacter jejuni and ca ...201121212120
influence of tetracycline resistance on the transport of manure-derived escherichia coli in saturated porous media.in this research, tetracycline resistant (tet(r)) and tetracycline susceptible (tet(s)) escherichia coli isolates were retrieved from dairy manure and the influence of tetracycline resistance on the transport of e. coli in saturated porous media was investigated through laboratory column transport experiments. experimental results showed that tet(r)e. coli strains had higher mobility than the tet(s) strains in saturated porous media. measurements of cell surface properties suggested that tet(r)e ...201021211813
survival of escherichia coli, enterococci, and campylobacter spp. in sheep feces on pastures.the survival of enteric bacteria in 10 freshly collected sheep fecal samples on pastures was measured in each of four seasons. ten freshly collected feces were placed on pasture, and concentrations of escherichia coli, enterococci, and campylobacter spp. were monitored until exhaustion of the fecal samples. in all four seasons, there was an increase in enterococcal concentrations by up to 3 orders of magnitude, with peak concentrations recorded between 11 and 28 days after deposition. e. coli co ...201121239546
persistence of free-living protozoan communities across rearing cycles in commercial poultry houses.the introduction and survival of zoonotic bacterial pathogens in poultry farming have been linked to bacterial association with free-living protozoa. to date, however, no information is available on the persistence of protozoan communities in these environments across consecutive rearing cycles and how it is affected by farm- and habitat-specific characteristics and management strategies. we therefore investigated the spatial and temporal dynamics of free-living protozoa in three habitats (pipel ...201121239551
influence of tetracycline resistance on the transport of manure-derived escherichia coli in saturated porous media.in this research, tetracycline resistant (tet(r)) and tetracycline susceptible (tet(s)) escherichia coli isolates were retrieved from dairy manure and the influence of tetracycline resistance on the transport of e. coli in saturated porous media was investigated through laboratory column transport experiments. experimental results showed that tet(r)e. coli strains had higher mobility than the tet(s) strains in saturated porous media. measurements of cell surface properties suggested that tet(r)e ...201021211813
do patients with recurrent episodes of campylobacteriosis differ from those with a single disease event?although campylobacter is the leading cause of reported bacterial gastro-enteritis in industrialized countries, little is known on its recurrence. the objective of this study is to describe the risk and the patient characteristics of recurrent episodes of human campylobacteriosis reported in quebec.201121226938
taxonomy, epidemiology, and clinical relevance of the genus arcobacter.the genus arcobacter, defined almost 20 years ago from members of the genus campylobacter, has become increasingly important because its members are being considered emergent enteropathogens and/or potential zoonotic agents. over recent years information that is relevant for microbiologists, especially those working in the medical and veterinary fields and in the food safety sector, has accumulated. recently, the genus has been enlarged with several new species. the complete genomes of arcobacte ...201121233511
neurologic symptoms associated with raising poultry and swine among participants in the agricultural health study.guillain-barré syndrome (gbs) is the leading cause of acute peripheral neuropathy worldwide, often associated with recent foodborne infection with campylobacter jejuni. in this cross-sectional analysis of data from the agricultural health study, we tested whether swine and poultry exposure were associated with increased prevalence of gbs-like neurologic symptoms.201121270654
campylobacter troglodytis sp. nov., isolated from feces of human-habituated wild chimpanzees (pan troglodytes schweinfurthii) in tanzania.the transmission of simian immunodeficiency and ebola viruses to humans in recent years has heightened awareness of the public health significance of zoonotic diseases of primate origin, particularly from chimpanzees. in this study, we analyzed 71 fecal samples collected from 2 different wild chimpanzee (pan troglodytes) populations with different histories in relation to their proximity to humans. campylobacter spp. were detected by culture in 19/56 (34%) group 1 (human habituated for research ...201121278267
prevalence of putative virulence markers in campylobacter jejuni and campylobacter coli isolated from hospitalized children, raw chicken, and raw beef in tehran, iran.the incidence of the virulence-associated genes cdta, cdtb, cdtc, cadf, dnaj, racr, and plda has been investigated in campylobacter jejuni and campylobacter coli collected from raw chicken and beef from retailers in tehran, iran, and from hospitalized children (age, =14 years) suffering from diarrhea. campylobacter spp. were collectively identified by morphological and biochemical methods. campylobacter jejuni and c. coli were discriminated from other campylobacter spp. by amplification of a spe ...201121326356
a charcoal- and blood-free enrichment broth for isolation and pcr detection of campylobacter jejuni and campylobacter coli in chicken.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 form ...201121333141
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