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ribotyping characterisation of campylobacter isolates randomly collected from different sources in italy.in this study the potential for using the automated psti ribotyping as a primary library typing method to survey campylobacter and for identification of two thermophilic campylobacter jejuni and campylobacter coli species was evaluated. a total of 158 isolates randomly collected in italy from different sources were analyzed. a large percentage of chicken (28%), turkey (27%) and turkey meat (25%) isolates shared their ribotyping profiles (ribotypes) with those of humans, whereas the swine isolate ...200314522511
applicability of a rapid duplex real-time pcr assay for speciation of campylobacter jejuni and campylobacter coli directly from culture plates.a rapid duplex real-time polymerase chain reaction (pcr) assay for speciation of campylobacter jejuni and campylobacter coli using the abi prism 7700 sequence detection system (applied biosystems) was developed based on two of the genes used in a conventional multiplex pcr. a rapid turnaround time of 3 h was achieved with the use of boiled cell lysates. applicability of the assay was tested with 6015 random campylobacter strains referred to the campylobacter reference unit, with 97.6% being iden ...200314680705
usefulness of the microseq 500 16s ribosomal dna-based bacterial identification system for identification of clinically significant bacterial isolates with ambiguous biochemical profiles.due to the inadequate automation in the amplification and sequencing procedures, the use of 16s rrna gene sequence-based methods in clinical microbiology laboratories is largely limited to identification of strains that are difficult to identify by phenotypic methods. in this study, using conventional full-sequence 16s rrna gene sequencing as the "gold standard," we evaluated the usefulness of the microseq 500 16s ribosomal dna (rdna)-based bacterial identification system, which involves amplifi ...200312734240
sequence analysis of a cryptic plasmid pcj419 from campylobacter jejuni and construction of an escherichia coli-campylobacter shuttle vector.a small cryptic plasmid, pcj419, was identified in a human clinical isolate of campylobacter jejuni, cloned and sequenced. pcj419 is a circular molecule of 4013 bp with a g+c content of 27.1%. the products of four open reading frames (orfs) share significant sequence similarity with putative proteins from known c. jejuni and campylobacter coli plasmids. orf-1 encodes a putative mobilisation protein (mob). orf-2 and orf-3 encode proteins that have high identity to putative repa and repb proteins, ...200312932741
in vitro activities of new fluoroquinolones against campylobacter jejuni and campylobacter coli isolates obtained from humans in 1980 to 1982 and 1997 to 2001.the antibacterial activities of three newly developed fluoroquinolones (gatifloxacin, levofloxacin, and moxifloxacin) against a total of 307 gastrointestinal human isolates of campylobacter jejuni and campylobacter coli collected during 1980 to 1982 and 1997 to 2001 were examined and compared to those of ciprofloxacin and the unrelated antibacterial agents, clarithromycin, erythromycin, and tetracycline by using the agar plate dilution method. all of the fluoroquinolones exhibited a good activit ...200312936999
development of a sensitive dna microarray suitable for rapid detection of campylobacter spp.campylobacter is the most common cause of human acute bacterial gastroenteritis worldwide, widely distributed and isolated from human clinical samples as well as from many other different sources. to comply with the demands of consumers for food safety, there is a need for development of a rapid, sensitive and specific detection method for campylobacter. in this study, we present the development of a novel sensitive dna-microarray based detection method, evaluated on campylobacter and non-campyl ...200312944122
comparison of the bax system with a multiplex pcr method for simultaneous detection and identification of campylobacter jejuni and campylobacter coli in environmental samples.the campylobacter detection is performed by conventional culture methods and the identification of campylobacter jejuni and campylobacter coli is principally based on the hippurate hydrolysis test. the two major drawbacks of this biochemical test for species identification include the inconsistency of the results and the presence of atypical strains, which can lead to the misidentification of an isolate. as an alternative, multiplex polymerase chain reaction (mpcr) protocols for the simultaneous ...200314527799
microarray-based identification of thermophilic campylobacter jejuni, c. coli, c. lari, and c. upsaliensis.dna microarrays are an excellent potential tool for clinical microbiology, since this technology allows relatively rapid identification and characterization of microbial and viral pathogens. in the present study, an oligonucleotide microarray was developed and used for the analysis of thermophilic campylobacter spp., the primary food-borne pathogen in the united states. we analyzed four campylobacter species: campylobacter jejuni, c. coli, c. lari, and c. upsaliensis. our assay relies on the pcr ...200312958228
application of host-specific bacteriophages to the surface of chicken skin leads to a reduction in recovery of campylobacter jejuni.retail poultry products are widely purported as the major infection vehicle for human campylobacteriosis. numerous intervention strategies have sought to reduce campylobacter contamination on broiler carcasses in the abattoir. this study reports the efficacy of bacteriophage in reducing the number of recoverable campylobacter jejuni cells on artificially contaminated chicken skin.200314532096
antimicrobial resistance in campylobacter from broilers: association with production type and antimicrobial use.the isolation and antimicrobial resistance of campylobacter jejuni and campylobacter coli strains from broilers arriving in french slaughterhouses, were analysed according to production types (i.e. standard, export or free-range) and antimicrobial (i.e. coccidiostats, growth promoters or therapeutic agents) administration in flocks. prevalence was 56.6% in standard, 51.3% in export and 80.0% in free-range broilers. three hundred and ninety-three strains were identified. two-thirds of the strains ...200314559174
deletion of helicobacter pylori vacuolating cytotoxin gene by introduction of directed mutagenesis.to construct a vaca-knockout helicobacter pylori mutant strain, whose only difference from the wild strain is its disrupted vaca gene.200314562387
pcr-restriction fragment length polymorphism analysis for detection of point mutations associated with macrolide resistance in campylobacter spp.a 23s rrna gene fragment in domain v was sequenced from 30 clinical isolates of campylobacter spp., including 22 resistant to macrolides. two point mutations associated with erythromycin resistance were identified at positions 2074 and 2075 on the 23s rrna gene (homologous to a2142c and a2143g mutations in helicobacter pylori) in which an adenine residue is also replaced with a cytosine and a guanine residue, respectively. a combined pcr-restriction fragment length polymorphism technique was dev ...200312604552
rapid detection of campylobacter coli, c. jejuni, and salmonella enterica on poultry carcasses by using pcr-enzyme-linked immunosorbent assay.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, campylobacter ...200312788755
a real-time pcr assay for the detection of campylobacter jejuni in foods after enrichment culture.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 contami ...200312620820
detection and typing of campylobacter jejuni and campylobacter coli and analysis of indicator organisms in three waterborne outbreaks in finland.waterborne outbreaks associated with contamination of drinking water by campylobacter jejuni are rather common in the nordic countries sweden, norway, and finland, where in sparsely populated districts groundwater is commonly used without disinfection. campylobacters, escherichia coli, or other coliforms have rarely been detected in potential sources. we studied three waterborne outbreaks in finland caused by c. jejuni and used sample volumes of 4,000 to 20,000 ml for analysis of campylobacters ...200312620821
comparison of broth microdilution, e test, and agar dilution methods for antibiotic susceptibility testing of campylobacter jejuni and campylobacter coli.a standardized broth microdilution method was compared to the e test and an agar dilution method for the antimicrobial susceptibility testing of campylobacter jejuni and c. coli isolates. a group of 47 human clinical isolates, 37 isolates from retail poultry, and 29 isolates from living turkeys (total, 113 isolates) was included in the study. these encompassed 92 c. jejuni and 21 c. coli strains. the mics of six antimicrobial agents were determined by the broth microdilution and e test methods, ...200312624030
molecular typing of campylobacter jejuni and campylobacter coli. 200312624086
antimicrobial resistance of campylobacter jejuni and campylobacter coli isolates from broiler chickens isolated at an irish poultry processing plant.the antibiotic susceptibility of campylobacter jejuni and campylobacter coli isolates from broiler chickens were determined in order to evaluate the level of antibiotic resistance of campylobacter species in the irish poultry industry.200312680938
helicobacter pylori preferentially induces interleukin 12 (il-12) rather than il-6 or il-10 in human dendritic cells.dendritic cells are potent antigen-presenting cells that are present in the gastrointestinal tract and are required for the induction of a th1 t-cell acquired immune response. since infection with the gastric pathogen helicobacter pylori elicits a th1 cell response, the interaction of these organisms with dendritic cells should reflect the th1 bias. we incubated h. pylori with cultured human dendritic cells and measured the cytokine induction profile, comparing the response to that induced by sa ...200312819109
survival and resuscitation of ten strains of campylobacter jejuni and campylobacter coli under acid conditions.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 recovered, even when enrichment media were used. viable cells, however, could be detected by a double-staining (5-cyano-2,3-ditolyl tetrazolium chloride [ctc]-4',6'-diamidino-2-phenylindole [dapi]) te ...200312514068
molecular assays for detection of human metapneumovirus.the recent description of the respiratory pathogen human metapneumovirus (hmpv) has highlighted a deficiency in current diagnostic techniques for viral agents associated with acute lower respiratory tract infections. we describe two novel approaches to the detection of viral rna by use of reverse transcriptase pcr (rt-pcr). the pcr products were identified after capture onto a solid-phase medium by hybridization with a sequence-specific, biotinylated oligonucleotide probe. the assay was applied ...200312517833
evaluation of 11 pcr assays for species-level identification of campylobacter jejuni and campylobacter coli.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 referen ...200312517869
real-time pcr assay for rapid and accurate detection of point mutations conferring resistance to clarithromycin in helicobacter pylori.the main cause of failure of helicobacter pylori eradication therapy is resistance to clarithromycin. the resistance is due to three point mutations in two positions on the 23s rrna (a2142c, a2142g, and a2143g). our aim was to develop a rapid and accurate method to detect these mutations directly on biopsy specimens. we developed a real-time pcr that included a simultaneous detection of the amplicons by hybridization of two probes labeled with lc-red and fluorescein by using the fluorescence res ...200312517879
prevalence of cytolethal distending toxin production in periodontopathogenic bacteria.cytolethal distending toxin (cdt) is a newly identified virulence factor produced by several pathogenic bacteria implicated in chronic infection. seventy three strains of periodontopathogenic bacteria were examined for the production of cdt by a hela cell bioassay and for the presence of the cdt gene by pcr with degenerative oligonucleotide primers, which were designed based on various regions of the escherichia coli and campylobacter cdtb genes, which have been successfully used for the identif ...200312682119
investigation of infectious agents associated with arthritis by reverse transcription pcr of bacterial rrna.in reactive and postinfectious arthritis the joints are generally sterile but the presence of bacterial antigens and nucleic acids has been reported. to investigate whether organisms traffic to affected joints in these conditions, we performed reverse transcription pcr using universal primers to amplify any bacterial 16s rrna sequences present in synovial fluid. bacterial sequences were detected in most cases, even after treatment of the synovial fluid with dnase, implying the presence of bacter ...200312716447
prevalence of multiple antibiotic resistance in 443 campylobacter spp. isolated from humans and animals.in view of recent findings that a multidrug efflux pump cmeabc exists in campylobacter jejuni, 391 c. jejuni and 52 campylobacter coli of human and animal origin were examined for a multidrug resistance phenotype.200312917241
investigation of infectious agents associated with arthritis by reverse transcription pcr of bacterial rrna.in reactive and postinfectious arthritis the joints are generally sterile but the presence of bacterial antigens and nucleic acids has been reported. to investigate whether organisms traffic to affected joints in these conditions, we performed reverse transcription pcr using universal primers to amplify any bacterial 16s rrna sequences present in synovial fluid. bacterial sequences were detected in most cases, even after treatment of the synovial fluid with dnase, implying the presence of bacter ...200312716447
phenotypic and genotypic differentiation of campylobacter spp. isolated from austrian broiler farms: a comparison.an identification scheme based on restriction fragment length polymorphism of polymerase chain reaction products (pcr-rflp) was developed to differentiate isolates of the genera campylobacter, arcobacter and helicobacter. based on the 16s rrna gene of these genera, pcr amplified a 1216-bp fragment. the amplicons were digested with the restriction enzymes rsai and ecorv. additional differentiation was obtained using a pcr-assay based on the hippuricase gene. genotyping was performed on several re ...200312745378
comparison of recovery methods for freeze-injured listeria monocytogenes, salmonella typhimurium, and campylobacter coli in cell suspensions and associated with pork surfaces.cells injured as a result of freezing, heating, and acidification treatments may not grow during conventional microbiological procedures owing to the presence of selective agents, compounds, or dyes in the media, impairing the cell's ability to repair itself and grow. injured cells can be recovered by combining selective and nonselective media into a single system. with such combinations, the diffusion of the selective compounds or dyes is controlled, allowing for the resuscitation of injured ce ...200312747688
pcr detection of seven virulence and toxin genes of campylobacter jejuni and campylobacter coli isolates from danish pigs and cattle and cytolethal distending toxin production of the isolates.to study the prevalence of seven virulence and toxin genes, and cytolethal distending toxin (cdt) production of campylobacter jejuni and c. coli isolates from danish pigs and cattle.200312752808
campylobacter--a tale of two protein glycosylation systems.post-translational glycosylation is a universal modification of proteins in eukarya, archaea and bacteria. two recent publications describe the first confirmed report of a bacterial n-linked glycosylation pathway in the human gastrointestinal pathogen campylobacter jejuni. in addition, an o-linked glycosylation pathway has been identified and characterized in c. jejuni and the related species campylobacter coli. both pathways have similarity to the respective n- and o-linked glycosylation proces ...200312781527
natural transformation of campylobacter jejuni requires components of a type ii secretion system.the human pathogen campylobacter jejuni is one of more than 40 naturally competent bacterial species able to import macromolecular dna from the environment and incorporate it into their genomes. however, in c. jejuni little is known about the genes involved in this process. we used random transposon mutagenesis to identify genes that are required for the transformation of this organism. we isolated mutants with insertions in 11 different genes; most of the mutants are affected in the dna uptake ...200312949093
toward an international standard for pcr-based detection of food-borne thermotolerant campylobacters: assay development and analytical validation.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 an ...200312957958
toward an international standard for pcr-based detection of food-borne thermotolerant campylobacters: validation in a multicenter collaborative trial.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 nontarget strains was tested, and the results were reported as the presence of a positive signal after gel electrophoresis. the overall inclusivity (sensitivity) was 93.7%, and the exclusivity (specificity) w ...200312957959
use of pcr for direct detection of campylobacter species in bovine feces.this study reports on the use of pcr to directly detect and distinguish campylobacter species in bovine feces without enrichment. inhibitors present in feces are a major obstacle to using pcr to detect microorganisms. the qiaamp dna stool minikit was found to be an efficacious extraction method, as determined by the positive amplification of internal control dna added to bovine feces before extraction. with nested or seminested multiplex pcr, campylobacter coli, c. fetus, c. hyointestinalis, and ...200312788747
production of cytolethal distending toxins by pathogenic escherichia coli strains isolated from human and animal sources: establishment of the existence of a new cdt variant (type iv).three types of cytolethal distending toxin (cdt), namely, cdt-i, cdt-ii, and cdt-iii, have been described in escherichia coli. using primers designed for the detection of sequences common to the cdtb genes, we analyzed by pcr a set of 21 cdt-producing e. coli strains of intestinal and extraintestinal origins isolated from human and different animal species in several european countries and in the united states. on the basis of the existing differences in the cdtb genes, cdt-i-, cdt-ii-, and cdt- ...200312958258
molecular characterization of fluoroquinolone-resistant campylobacter spp. isolated from poultry.campylobacteriosis, an infectious disease caused by campylobacter jejuni and campylobacter coli, is treated by fluoroquinolone antibiotics in clinical practices. however, use of these drugs in animal husbandry may select for fluoroquinolone-resistant campylobacters and, thereby, compromise the clinical treatment of infection. in this study, 21 fluoroquinolone-resistant campylobacters were isolated from poultry samples. morphological and biochemical characteristics indicated that 19 isolates were ...200312619802
campylobacter coli - an important foodborne pathogen.campylobacters are the most common bacterial cause of infectious intestinal disease (iid) in temperate countries. c. jejuni is the predominant cause of campylobacter iid, but the impact of other, less prevalent species has largely been ignored. here, we present estimates of the burden of indigenously acquired foodborne disease (ifd) due to campylobacter coli, the second most common cause of human campylobacteriosis.200312850159
detection and enumeration of campylobacter jejuni and campylobacter coli by indirect impedimetry with an oxygen scavenging system.six strains of campylobacter jejuni and campylobacter coli were shown to grow in a variety of media, but, with one exception, they were unable to produce sufficient change in the electrical properties of the medium to allow their detection by impedance monitoring. with the use of an indirect method based on absorption of evolved carbon dioxide and a medium containing the oxygen scavenger oxyrase, all strains were detectable, and correlations between time to detection and the logarithm of the ino ...200314503735
inhibition of helicobacter pylori growth in vitro by bulgarian propolis: preliminary report.bee glue (propolis) possesses antimicrobial, anti-inflammatory, anaesthetic and immunostimulating activities. the aim of the study was to evaluate the inhibitory effect of bulgarian propolis on helicobacter pylori growth in vitro. activity of 30% ethanolic extract of propolis (eep) against 38 clinical isolates of h. pylori was evaluated by using the agar-well diffusion method. ethanol was used as a control. in addition, the effect of propolis on the growth of 26 h. pylori and 18 campylobacter st ...200312721318
antimicrobial-resistant campylobacter species from retail raw meats.the antimicrobial susceptibilities of 378 campylobacter isolates were determined. resistance to tetracycline was the most common (82%), followed by resistance to doxycycline (77%), erythromycin (54%), nalidixic acid (41%), and ciprofloxacin (35%). campylobacter coli displayed significantly higher rates of resistance to ciprofloxacin and erythromycin than campylobacter jejuni, and campylobacter isolates from turkey meat showed a greater resistance than those from chicken meat.200312732579
antimicrobial resistance of campylobacter jejuni subsp. jejuni strains isolated from humans in 1998 to 2001 in montréal, canada.the rates of resistance of 51 to 72 human strains of campylobacter jejuni subsp. jejuni isolated annually from 1998 to 2001 in montréal, québec, canada, varied from 1 to 12% for erythromycin, 43 to 68% for tetracycline, and 10 to 47% for ciprofloxacin. in the last years of the study, there was a significant increase in the rate of resistance to ciprofloxacin (p = 0.00003) but not in the rate of resistance to erythromycin (p = 0.056) or tetracycline (p = 0.095) compared to the rate obtained in th ...200312760892
the iron-responsive regulator fur is transcriptionally autoregulated and not essential in neisseria meningitidis.fur is a well-known iron-responsive repressor of gene transcription, which is used by many bacteria to respond to the low-iron environment that pathogens encounter during infection. the fur gene in neisseria meningitidis has been described as an essential gene that may regulate a broad array of genes. we succeeded in obtaining an n. meningitidis mutant with the fur gene knocked out and used it to undertake studies of fur-mediated iron regulation. we show that expression of both fur and the trans ...200314526014
specific detection of arcobacter and campylobacter strains in water and sewage by pcr and fluorescent in situ hybridization.the aim of this study was to evaluate pcr and fluorescent in situ hybridization (fish) techniques for detecting arcobacter and campylobacter strains in river water and wastewater samples. both 16s and 23s rrna sequence data were used to design specific primers and oligonucleotide probes for pcr and fish analyses, respectively. in order to assess the suitability of the methods, the assays were performed on naturally and artificially contaminated samples and compared with the isolation of cells on ...200312571045
epidemiology and antimicrobial susceptibilities of 111 campylobacter fetus subsp. fetus strains isolated in québec, canada, from 1983 to 2000.the epidemiology and antimicrobial susceptibilities of 111 campylobacter fetus subsp. fetus strains isolated from 103 patients from 1983 to 2000 in québec, canada, were determined. the median number of patients infected annually with this bacteria was seven, with an incidence of 0.1 per 100,000 population. the male-to-female ratio was 1.1 to 1.0. the patients originated from 13 of the 18 québec socioeconomic regions. the age range of the patients was 6 months to 90 years old, 53% being > or = 70 ...200312517895
the gam protein of bacteriophage mu is an orthologue of eukaryotic ku.mu bacteriophage inserts its dna into the genome of host bacteria and is used as a model for dna transposition events in other systems. the eukaryotic ku protein has key roles in dna repair and in certain transposition events. here we show that the gam protein of phage mu is conserved in bacteria, has sequence homology with both subunits of ku, and has the potential to adopt a similar architecture to the core dna-binding region of ku. through biochemical studies, we demonstrate that gam and the ...200312524520
characterization of helicobacter pylori nickel metabolism accessory proteins needed for maturation of both urease and hydrogenase.previous studies demonstrated that two accessory proteins, hypa and hypb, play a role in nickel-dependent maturation of both hydrogenase and urease in helicobacter pylori. here, the two proteins were purified and characterized. hypa bound two ni(2+) ions per dimer with positive cooperativity (hill coefficient, approximately 2.0). the dissociation constants k(1) and k(2) for ni(2+) were 58 and 1.3 microm, respectively. studies on purified site-directed mutant proteins in each of the five histidin ...200312533448
the iron-binding protein dps confers hydrogen peroxide stress resistance to campylobacter jejuni.we identified and characterized the iron-binding protein dps from campylobacter jejuni. electron microscopic analysis of this protein revealed a spherical structure of 8.5 nm in diameter, with an electron-dense core similar to those of other proteins of the dps (dna-binding protein from starved cells) family. cloning and sequencing of the dps-encoding gene (dps) revealed that a 450-bp open reading frame (orf) encoded a protein of 150 amino acids with a calculated molecular mass of 17,332 da. ami ...200312533477
in vivo tracking of campylobacter jejuni by using a novel recombinant expressing green fluorescent protein.campylobacter jejuni is a leading cause of food-borne disease in developed countries. the goal of this study was to develop a plasmid-based reporter system with green fluorescent protein (gfp) to facilitate the study of c. jejuni in a variety of niches. c. jejuni transformants harboring the pmek91 gfp gene (gfp)-containing vector were readily detectable by both fluorescence microscopy and flow cytometry. given the ease of detecting these organisms, additional experiments were performed in which ...200312732559
birds, migration and emerging zoonoses: west nile virus, lyme disease, influenza a and enteropathogens.wild birds are important to public health because they carry emerging zoonotic pathogens, either as a reservoir host or by dispersing infected arthropod vectors. in addition, bird migration provides a mechanism for the establishment of new endemic foci of disease at great distances from where an infection was acquired. birds are central to the epidemiology of west nile virus (wnv) because they are the main amplifying host of the virus in nature. the initial spread of wnv in the u.s. along the ea ...200315931279
comparison of campylobacter isolates from poultry and humans: association between in vitro virulence properties, biotypes, and pulsed-field gel electrophoresis clusters.the in vitro virulence properties of 197 temporally and geographically related campylobacter isolates from chicken broilers and humans were compared. comparisons of the virulence properties associated with genotypes and biotypes were made. all isolates adhered to, and 63% invaded, int-407 cells, whereas 13% were cytotoxic for cho cells. cho cell-cytotoxic extracts were also cytotoxic for int-407 cells, but the sensitivity for vero cells was variable. the proportion of isolates demonstrating a hi ...200314532099
a survey of campylobacter species shed in faeces of beef cattle using polymerase chain reaction.a polymerase chain reaction (pcr)-based survey of campylobacters associated with faeces collected from 382 beef cattle was undertaken. to ensure the removal of pcr inhibitors present in faeces and determine if adequate extraction was achieved, faeces were seeded with internal control dna (i.e., dna designed to amplify with the campylobacter genus primer set, but provide a smaller amplicon) before the extraction procedure. in only two samples (0.5%) were the internal control or campylobacter genu ...200314735214
antimicrobial resistance in campylobacter jejuni and campylobacter coli strains isolated in 1991 and 2001-2002 from poultry and humans in berlin, germany.the susceptibilities of 430 campylobacter jejuni strains and 79 c. coli strains to six antimicrobial agents were tested and analyzed. the two sets of strains originated from retail market chicken and turkey samples and from humans, respectively, in berlin, germany. two groups of isolates, one dating from 1991 and the other dating from 2001-2002, were tested. of the campylobacter sp. isolates recovered from humans in 2001-2002, 45.1% were resistant to ciprofloxacin, 37.8% were resistant to tetrac ...200314638490
antimicrobial susceptibility trends in campylobacter jejuni and campylobacter coli isolated from a rural egyptian pediatric population with diarrhea.comparative and trend analysis was conducted on annual prevalence of antimicrobial susceptibility among campylobacter jejuni and campylobacter coli recovered from rural egyptian children from 1995 through 2000. c. jejuni and c. coli demonstrated significant decreasing trends in ciprofloxacin susceptibility over the study period (p < 0.001 for both). in general, c. coli demonstrated a higher degree of susceptibility than c. jejuni, however, there was no statistical difference (p = 0.2) comparing ...200314711482
routes for campylobacter contamination of poultry meat: epidemiological study from hatchery to slaughterhouse.from april 1998 to march 2000, 18 broiler flocks were followed from the hatchery to the slaughterhouse. campylobacter was not found in the hatchery, 1-day-old chicks or in the rearing house before the arrival of the chicks. the infection of broiler flocks increased continuously during the rearing time, with a total of seven positive flocks at the end of rearing. farms with campylobacter-positive broilers were characterized by the circulation of campylobacter in the environment (puddles, dung hil ...200314959785
risk factors for sporadic campylobacter jejuni infections in rural michigan: a prospective case-control study.this case-control study investigated risk factors for campylobacteriosis in a rural population. exposure to live farm animals was hypothesized to increase the risk for campylobacter jejuni enteritis.200314652344
fluoroquinolone-resistant campylobacter in animal reservoirs: dynamics of development, resistance mechanisms and ecological fitness.thermophilic campylobacter species, including campylobacter jejuni and campylobacter coli, are responsible for foodborne campylobacteriosis in humans and are increasingly resistant to fluoroquinolone (fq) antimicrobials. the therapeutic use of fq antimicrobial agents in food animal production, particularly in poultry, has become a concern for public health, because the practice may promote the emergence of fq-resistant campylobacter that can be transmitted to humans through the food chain. recen ...200315134291
cecal colonization of chicks by bovine-derived strains of campylobacter.campylobacter jejuni and campylobacter coli strains were isolated from feces of dairy cattle at farms with no known problem due to campylobacteria. farms were located in the northeast, desert southwest, and pacific west. twenty isolates were identified by ribotyping with a riboprinter. the ability of these bovine isolates to colonize the ceca of chicks was determined by challenge inoculation and reisolation of the challenge strain from the ceca at 1 and 2 wk after challenge. isolates recovered f ...200314708992
characterization of a haemolytic phospholipase a(2) activity in clinical isolates of campylobacter concisus.a membrane-bound, haemolytic phospholipase a(2) (pla(2)) activity was detected in clinical strains of campylobacter concisus isolated from children with gastroenteritis. the clinical strains were assigned into two molecular groups (genomospecies) based on pcr amplification of their 23s rdna. this calcium-dependent, heat-stable, haemolytic pla(2) activity was detected in strains from both genomospecies. a crude haemolysin extract (che) was initially prepared from cellular outer-membrane proteins ...200415150326
prevalence and antimicrobial resistance of campylobacter coli isolated from fattening pigs in france.campylobacter are a leading cause of human diarrhea. the usual source of infection is contaminated food, particularly poultry but pork has also been described. the veterinary use of antimicrobial drugs has been suggested to be largely responsible for resistance in human isolates of this zoonotic pathogen. a study was carried out to investigate the occurrence and antimicrobial resistance of campylobacter isolated from french fattening pigs. from march 1998 to june 1999, stomach samples were colle ...200415172691
in vitro inhibitory activity of chinese leek extract against campylobacter species.three aqueous extracts of dietary materials of chinese leek (leek flower stem, soft leek and green leek) and two other reference extracts of allium plants were used to investigate the antibacterial effects against campylobacter jejuni and campylobacter coli. all the tested strains of campylobacter species were isolated from chickens. the minimum concentration of aqueous leek extracts (ales) required to inhibit the bacterial growth was 2.0 mg/ml. among the plant extracts tested, ales shared the l ...200415193803
genotyping of broiler-originated campylobacter jejuni and campylobacter coli isolates using fla typing and random amplified polymorphic dna methods.liver and intestine samples taken from 200 broilers at 20 flocks were inoculated onto preston enrichment broth and agar for selective isolation of campylobacter jejuni and campylobacter coli. the isolates were identified by both conventional and polymerase chain reaction (pcr) methods. campylobacter spp. were identified in 102 of 400 samples (200 liver and 200 intestine), 57 (14.25%) of which were identified as c. jejuni and 45 (11.25%) as c. coli. pcr-restriction fragment length polymorphism (r ...200415193806
swimming and campylobacter infections.a matched case-control study was conducted to study risk factors for domestically acquired sporadic campylobacter infections in finland. swimming in natural sources of water was a novel risk factor. eating undercooked meat and drinking dug-well water were also independent risk factors for campylobacter infection.200415496253
relapsing cellulitis associated with campylobacter coli bacteremia in an agammaglobulinemic patient.campylobacter coli rarely causes bacteremia or extraintestinal infection. we report herein a case of agammaglobulinemia in which cellulitis associated with c. coli bacteremia relapsed after a disease-free interval of >5 years. pulsed field gel electrophoresis revealed that the organisms in this patient were genetically identical, suggesting a latent c. coli infection.200415194845
adaptation of campylobacter jejuni nctc11168 to high-level colonization of the avian gastrointestinal tract.the genome sequence of the human pathogen campylobacter jejuni nctc11168 has been determined recently, but studies on colonization and persistence in chickens have been limited due to reports that this strain is a poor colonizer. experimental colonization and persistence studies were carried out with c. jejuni nctc11168 by using 2-week-old light sussex chickens possessing an acquired natural gut flora. after inoculation, nctc11168 initially colonized the intestine poorly. however, after 5 weeks ...200415213117
molecular characterization of benzimidazole resistance in helicobacter pylori.a family of benzimidazole derivatives (bi) was shown to possess potent and selective activity against helicobacter pylori, although the precise cellular target of the bis is unknown. spontaneous h. pylori mutants were isolated as resistant to a representative bi (compound a). genomic dna was isolated from a bi-resistant mutant, transformed into a bi-sensitive strain, and found to be sufficient to confer bi resistance. the resistance determinant was localized to a 17-kb clone after screening a la ...200415215104
iron acquisition and regulation in campylobacter jejuni.iron affects the physiology of bacteria in two different ways: as a micronutrient for bacterial growth and as a catalyst for the formation of hydroxyl radicals. in this study, we used dna microarrays to identify the c. jejuni genes that have their transcript abundance affected by iron availability. the transcript levels of 647 genes were affected after the addition of iron to iron-limited c. jejuni cells. several classes of affected genes were revealed within 15 min, including immediate-early re ...200415231804
longitudinal study of campylobacter jejuni bacteriophages and their hosts from broiler chickens.a longitudinal study of bacteriophages and their hosts was carried out at a broiler house that had been identified as having a population of campylobacter-specific bacteriophages. cloacal and excreta samples were collected from three successive broiler flocks reared in the same barn. campylobacter jejuni was isolated from each flock, whereas bacteriophages could be isolated from flocks 1 and 2 but were not isolated from flock 3. the bacteriophages isolated from flocks 1 and 2 were closely relate ...200415240258
identification of bacterial populations in dairy wastewaters by use of 16s rrna gene sequences and other genetic markers.hydraulic flush waste removal systems coupled to solid/liquid separators and circulated treatment lagoons are commonly utilized to manage the large amounts of animal waste produced on high-intensity dairy farms. although these systems are common, little is known about the microbial populations that inhabit them or how they change as they traverse the system. using culture-based and non-culture-based methods, we characterized the microbial community structure of manure, water from the separator p ...200415240310
campylobacter insulaenigrae sp. nov., isolated from marine mammals.phenotypic and phylogenetic studies were performed on four campylobacter-like organisms recovered from three seals and a porpoise. comparative 16s rrna gene sequencing studies demonstrated that the organisms represent a hitherto unknown subline within the genus campylobacter, associated with a subcluster containing campylobacter jejuni, campylobacter coli and campylobacter lari. dna-dna hybridization studies confirmed that the bacteria belonged to a single species, for which the name campylobact ...200415545485
multiplex pcr assay for detection of streptococcus suis species and serotypes 2 and 1/2 in tonsils of live and dead pigs.a pcr assay was developed for the detection of streptococcus suis serotypes 2 and 1/2. this multiplex pcr is based on the amplification of the gene coding for 16s rrna of s. suis and on the amplification of the cps2j gene coding for the capsule of s. suis serotypes 2 and 1/2. an internal control was constructed and added in this test to monitor the efficiency of amplification in each reaction. to evaluate the specificity of the test, 31 strains of other bacterial species related to s. suis or is ...200415243078
characterization of plasmid-mediated apha-3 kanamycin resistance in campylobacter jejuni.a total of 254 isolates of campylobacter jejuni and three isolates of campylobacter coli, isolated from sweden, canada, and egypt, were screened for kanamycin resistance. eight strains of c. jejuni contained large plasmids that carried the apha-3 kanamycin-resistance marker. in six plasmids, the apha-3 gene was located downstream of an apparent insertion sequence, designated is607*, which showed a considerable similarity to is607, characterized on the chromosome of some helicobacter pylori strai ...200415256024
campylobacter colonization of sibling turkey flocks reared under different management conditions.uncertainty exists concerning the key factors contributing to campylobacter colonization of poultry, especially the possible role of vertical transmission from breeder hens to young birds. a longitudinal study of campylobacter colonization was performed in two sibling pairs of turkey flocks (four flocks total). each pair of sibling flocks shared breeder hen populations and was obtained from the same hatchery. one flock of each pair was grown on a commercial farm, and the other was grown in an in ...200415270502
campylobacter prevalence in lactating dairy cows in the united states.the objective of the present study was to determine the prevalence of intestinal campylobacter in lactating dairy cows from various regions of the united states. participating commercial dairy farms were chosen at random and were part of a national survey to determine e. coli o157:h7 and salmonella prevalence in dairy cows. farms had no previous history of campylobacter problems. fecal samples were collected rectally from 720 cows on farms in the northeast (four farms), in the desert southwest ( ...200415270504
characterization of lactobacillus coryniformis dsm 20001t surface protein cpf mediating coaggregation with and aggregation among pathogens.phenotypic characterization of aggregation phenotypes of lactobacillus coryniformis revealed that strain dsm 20001t coaggregated with escherichia coli k88, campylobacter coli, and campylobacter jejuni but not with other human pathogens. in addition, cells of these pathogens aggregated in the presence of the spent culture supernatant (scs) of strain dsm 20001t. cells of e. coli k88 remained viable in the coaggregates and aggregates for up to 24 h. both coaggregation and aggregation (co/aggregatio ...200415574903
a real-time multiplexed pcr assay for rapid detection and differentiation of campylobacter jejuni and campylobacter coli.campylobacter species are the leading agents of bacterial gastroenteritis worldwide. c. jejuni and c. coli together are responsible for more than 95% of all cases of campylobacter-induced diarrheal disease in the united states. detection of campylobacteria in clinical samples by conventional culture is problematic and slow due to their complex taxonomy, fastidious growth requirements, and biochemical inertness. the current study describes a rapid, sensitive, and specific real-time polymerase cha ...200415271389
helicobacter pylori induces apoptosis of human monocytes but not monocyte-derived dendritic cells: role of the cag pathogenicity island.monocytes are circulating precursors of the dendritic cell subset, professional antigen-presenting cells with a unique ability to initiate the innate and adaptive immune response. in this study, we have investigated the effects of wild-type helicobacter pylori strains and their isogenic mutants with mutations in known bacterial virulence factors on monocytes and monocyte-derived dendritic cells. we show that h. pylori strains induce apoptosis of human monocytes by a mechanism that is dependent o ...200415271906
occurrence of campylobacter in retail foods in ireland.a surveillance study was carried out to determine the prevalence of campylobacter in a range of retail foods purchased in three irish cities over a 20-month period between march 2001 and october 2002. in total 2391 food samples were analysed during this period. campylobacter was isolated from 444 raw chicken (49.9%), 33 turkey (37.5%) and 11 duck samples (45.8%). lower isolation rates of 7/221 (3.2%), 10/197 (5.1%) and 31/262 (11.8%) were observed for raw beef, pork and lamb, respectively. one s ...200415282123
the pattern and kinetics of substrate metabolism of campylobacter jejuni and campylobacter coli.the main aim was to investigate the patterns and kinetics of substrate oxidation by campylobacter jejuni and c. coli.200415287872
role of an inducible single-domain hemoglobin in mediating resistance to nitric oxide and nitrosative stress in campylobacter jejuni and campylobacter coli.campylobacter jejuni expresses two hemoglobins, each of which exhibits a heme pocket and structural signatures in common with vertebrate and plant globins. one of these, designated cgb, is homologous to vgb from vitreoscilla stercoraria and does not possess the reductase domain seen in the flavohemoglobins. a cgb-deficient mutant of c. jejuni was hypersensitive to nitrosating agents (s-nitrosoglutathione [gsno] or sodium nitroprusside) and a nitric oxide-releasing compound (spermine nonoate). th ...200415292134
antimicrobial susceptibilities of campylobacter isolated from food-producing animals on farms (1999-2001): results from the japanese veterinary antimicrobial resistance monitoring program.nationwide monitoring of antimicrobial resistance in campylobacter isolated from food-producing animals on farms was performed in japan. a total of 468 campylobacter isolates were obtained during the period from june 1999 to march 2001. campylobacter species showed high frequencies of resistance to oxytetracycline and dihydrostreptomycin. the frequencies of resistance in campylobacter coli to aminoglycosides, macrolides, tetracycline and quinolones were higher than those in campylobacter jejuni ...200415325430
antigenicity of the campylobacter coli cjaa protein produced by escherichia coli.immunogenic c. coli cjaa protein is a candidate for a chicken anti-campylobacter subunit vaccine. in order to enhance its immunogenicity an antigenic determinant of the cjaa protein was identified. thereafter, two copies of the antigenic epitope were cloned in tandem directly or with a flexible hinge between them. all experiments documented that the cjaa protein contained not only linear antigenic epitope/s but also conformational ones.200415330269
use of midi-fatty acid methyl ester analysis to monitor the transmission of campylobacter during commercial poultry processing.the presence of campylobacter spp. on broiler carcasses and in scald water taken from a commercial poultry processing facility was monitored on a monthly basis from january through june. campylobacter agar, blaser, was used to enumerate campylobacter in water samples from a multiple-tank scalder; on prescalded, picked, eviscerated, and chilled carcasses; and on processed carcasses stored at 4 degrees c for 7 or 14 days. the midi sherlock microbial identification system was used to identify campy ...200415330523
species identification by genotyping and determination of antibiotic resistance in campylobacter jejuni and campylobacter coli from humans and chickens in sweden.campylobacter is today the most common cause of human bacterial enteritis in sweden, as well as in most other industrialized countries. common sources of infection are undercooked chicken meat, unpasteurized milk and contaminated drinking water. one aim with our present study was to identify the species campylobacter jejuni and campylobacter coli strains from humans and chickens using a polymerase chain reaction/restriction enzyme analysis (pcr/rea) method, as well as traditional hippurate hydro ...200415364471
use of culture, pcr analysis, and dna microarrays for detection of campylobacter jejuni and campylobacter coli from chicken feces.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 jejun ...200415364980
campylobacter, from obscurity to celebrity.after its successful isolation from stools in the 1970s, campylobacter jejuni has rapidly become the most commonly recognised cause of bacterial gastroenteritis in man. reported cases of human campylobacteriosis represent only a small fraction of the actual number. in industrialised countries, the incidence of c. jejuni/campylobacter coli infections peaks during infancy, and again in young adults aged 15-44 years. acute self-limited gastrointestinal illness, characterised by diarrhoea, fever and ...200415373879
a european survey of antimicrobial susceptibility among zoonotic and commensal bacteria isolated from food-producing animals.to study antimicrobial resistance in zoonotic bacteria isolated from food animals in different countries using uniform methodology.200415375107
antimicrobial resistance among enteric pathogens.diarrhea is a major cause of morbidity and mortality in infants and children worldwide. use of antimicrobial therapy in children with bacterial diarrhea involves consideration of advantages and limitations of use of appropriate agents in the general population and in specific hosts. antimicrobial agents for bacterial diarrhea should be prescribed with an appreciation of limitations including antimicrobial resistance. studies from many countries have reported resistance to campylobacter jejuni, c ...200415185189
nucleotide sequences and comparison of two large conjugative plasmids from different campylobacter species.two large tetracycline resistance (tcr) plasmids have been completely sequenced, the ptet plasmid (45.2 kb) from campylobacter jejuni strain 81-176 and a plasmid pcc31 (44.7 kb) from campylobacter coli strain cc31 that was isolated from a human case of severe gastroenteritis in the uk. both plasmids are mosaic in structure, having homologues of genes found in a variety of different commensal and pathogenic bacteria, but nevertheless, showed striking similarities in dna sequence and overall gene ...200415470128
antimicrobial resistance of campylobacter jejuni and campylobacter coli isolates in japan. 200415499812
occurrence of campylobacter jejuni in pets living with human patients infected with c. jejuni.campylobacter jejuni was recovered from four dogs (11%) and four cats (33%) living with danish human patients infected with c. jejuni. pulsed-field gel electrophoresis (pfge) analysis revealed the occurrence of the same quinolone-resistant strain in a girl and her dog. c. jejuni isolates with closely related (>95% similarity) pfge profiles occurred in humans and pets from different danish counties.200415004120
[the role of certain campylobacter types in the etiology of enterocolitis].in recent decades, medical community has increasingly been calling attention to the importance of campylobacter as an disease-causing agent in humans. nowadays, 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 ...200415022385
emergence of fluoroquinolone resistance in the native campylobacter coli population of pigs exposed to enrofloxacin.the effect of a single 5 day enrofloxacin treatment on the native campylobacter coli population in conventionally weaned 5-week-old pigs was investigated.materials: twelve pigs were split into two groups of six: one group was treated with a therapeutic dose (15 mg/pig/day) of enrofloxacin and the other remained untreated to act as the control. campylobacter coli were isolated from faecal samples and tested for ciprofloxacin resistance by measuring mic values. mutations in the quinolone resistanc ...200415028665
sequence polymorphism in the glycosylation island and flagellins of pseudomonas aeruginosa.a genomic island consisting of 14 open reading frames, orfa to orfn was previously identified in pseudomonas aeruginosa strain pak and shown to be essential for glycosylation of flagellin. dna microarray hybridization analysis of a number of p. aeruginosa strains from diverse origins showed that this island is polymorphic. pcr and sequence analysis confirmed that many p. aeruginosa strains carry an abbreviated version of the island (short island) in which orfd, -e and -h are polymorphic and orfi ...200415028697
development of a multiplex microarray microsystem.a hybrid multiplex microarray microsystem has been developed that consists of 32 individually addressable array reaction chambers, supporting the use of multichannel pipettes for addition of up to 8 samples simultaneously. discrimination between campylobacter jejuni and campylobacter coli bacteria was observed in dna samples containing campylobacter spp., with the same specificity and sensitivity as when compared to a full-size microarray. the spinloaded multiplex microarray microsystem describe ...200415052357
use of the omp50 gene for identification of campylobacter species by pcr.we studied the prevalence of the omp50 gene and the omp50 protein in campylobacter strains. immunodetection assays and dna-dna hybridizations showed that most c. coli strains tested were negative and most c. jejuni and c. lari strains tested were positive. a pcr assay was developed, using the omp50 gene as a species-specific target. we propose a combination of a hippurate test and an omp50 assay to perform identification of campylobacter species.200415131219
characterization of campylobacter isolates recovered from clinically healthy pigs and from sporadic cases of campylobacteriosis in humans.campylobacter spp. were recovered from 660 (77.6%) of 850 swine cecal contents at the abattoir and from 24 (8.6%) of 278 specimens from sporadic cases of human diarrhea during the same period in the same geographical area. campylobacter coli represented 95.7% of campylobacter isolates recovered from pigs and 8.3% of those isolated from humans. genetic profiles were determined by pulsed-field gel electrophoresis (pfge) using kpni enzyme to characterize the isolates in combination with phenotypic ...200414968951
dual control of helicobacter pylori heat shock gene transcription by hspr and hrca.the hspr repressor regulates transcription of the groesl, hrca-grpe-dnak, and cbpa-hspr-orf operons of helicobacter pylori. here we show that two of the hspr-regulated operons, namely, the groesl and dnak operons, encoding the major cellular chaperone machineries are also regulated by the h. pylori homologue of the hrca repressor. similarly to the hspr mutation, deletion of the hrca gene also leads to complete derepression of the pgro and phrc promoters. the presence of both hspr and hrca is the ...200415126455
optimization of pcr-based methods for rapid detection of campylobacter jejuni, campylobacter coli and yersinia enterocolitica serovar 0:3 in wastewater samples.pcr-based methods were evaluated for their adequacy to assess the removal of pathogens from wastewater samples. for the development and optimization of the methods, samples were taken at two different sites from two different constructed wetlands. campylobacter jejuni/coli and yersinia enterocolitica serogroup 0:3 were selected as model pathogens and enterococcus faecalis as a standard microbiological indicator. the chosen pcr protocols were optimized for wastewater dna extracts in order to obta ...200414975667
multiplex pcr assay for differentiation of helicobacter felis, h. bizzozeronii, and h. salomonis.helicobacter felis, helicobacter bizzozeronii, and helicobacter salomonis are frequently found in the gastric mucous membrane of dogs and cats. these large spiral organisms are phylogenetically highly related to each other. their fastidious nature makes it difficult to cultivate them in vitro, hampering traditional identification methods. we describe here a multiplex pcr test based on the trna intergenic spacers and on the urease gene, combined with capillary electrophoresis, that allows discrim ...200415004062
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