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cytokine responses in primary chicken embryo intestinal cells infected with campylobacter jejuni strains of human and chicken origin and the expression of bacterial virulence-associated genes.campylobacter jejuni is a major cause of inflammatory diarrhoea in humans and is considered a commensal of the gastroenteric tract of the avian host. however, little is known about the interaction between c. jejuni and the avian host including the cytokine responses and the expression of the bacterial genes. we have investigated the invasiveness of primary chicken embryo intestinal cells (ceics) by c. jejuni strains of human and chicken origins and the production of pro-inflammatory cytokines as ...200818588667
host-pathogen interactions in campylobacter infections: the host perspective.campylobacter is a major cause of acute bacterial diarrhea in humans worldwide. this study was aimed at summarizing the current understanding of host mechanisms involved in the defense against campylobacter by evaluating data available from three sources: (i) epidemiological observations, (ii) observations of patients, and (iii) experimental observations including observations of animal models and human volunteer studies. analysis of available data clearly indicates that an effective immune syst ...200818625685
campylobacter-induced interleukin-8 secretion in polarized human intestinal epithelial cells requires campylobacter-secreted cytolethal distending toxin- and toll-like receptor-mediated activation of nf-kappab.campylobacter jejuni and campylobacter coli colonize and infect the intestinal epithelium and cause acute inflammatory diarrhea. the intestinal epithelium serves as a physical barrier to, and a sensor of, bacterial infection by secreting proinflammatory cytokines. this study examined the mechanisms for campylobacter-induced secretion of the proinflammatory chemokine interleukin-8 (il-8) by using polarized t84 human colonic epithelial cells as a model. c. jejuni increased the secretion of both il ...200818644884
the omp50 gene is transcriptionally controlled by a temperature-dependent mechanism conserved among thermophilic campylobacter species.the thermophilic campylobacters are enteropathogenic for humans. we recently showed that omp50 is a campylobacter species-specific porin produced in campylobacter jejuni and campylobacter lari but not in campylobacter coli. in the present study, we investigated regulation of the omp50 gene and found that its expression in c. jejuni was temperature-dependent, but independent of growth phase or medium viscosity. the use of rt-pcr and omp50::lacz fusions showed that growth temperature control occur ...200818485677
the haemophilus influenzae hmw1 adhesin is a glycoprotein with an unusual n-linked carbohydrate modification.the haemophilus influenzae hmw1 adhesin mediates adherence to respiratory epithelial cells, a critical early step in the pathogenesis of h. influenzae disease. in recent work, we demonstrated that hmw1 undergoes glycosylation. in addition, we observed that glycosylation of hmw1 is essential for hmw1 tethering to the bacterial surface, a prerequisite for hmw1-mediated adherence to host epithelium. in this study, we examined hmw1 proteolytic fragments by mass spectrometry, achieved 89% amino acid ...200818621734
diversity of pulsed-field gel electrophoresis profiles of campylobacter jejuni and campylobacter coli from broiler chickens in france.in 2003 to 2004, 26 free-range broilers flocks excreting campylobacter were studied for identification of campylobacter species and genotype diversity. seventeen flocks were sampled at the end of the indoor rearing period and 9 before departure to the slaughterhouse after access to an open area. out of 513 isolates, 315 were identified as campylobacter jejuni and 198 as campylobacter coli. pulsed-field gel electrophoresis analysis revealed 35 genotypes for c. jejuni and 43 genotypes for c. coli; ...200818648064
monoclonal antibody binding to the major outer membrane protein of campylobacter coli.campylobacter species are major enteric pathogens causing diarrhea illness in humans and animals. immunological tests are needed for accurate and rapid identification of c. coli, in conjunction with the use of standard biochemical tests. we initiated the creation of monoclonal antibodies (mabs) using whole c. coli cells as antigen. four positive clones were identified, namely mab2g6, mab3b9, mab4a10 and mab5b9. dot-blot assay and elisa revealed that only mab2g6 did not cross react with c. jejuni ...200818796306
occurrence and genotypes of campylobacter in broiler flocks, other farm animals, and the environment during several rearing periods on selected poultry farms.on 15 swiss poultry farms, broiler flocks, other farm animals, and the environment were examined during consecutive rearing periods to investigate the occurrence and genetic diversity of campylobacter. of the 5154 collected samples, 311 (6%) from 14 farms were campylobacter positive by culture. amongst the positive samples, 228 tested positive for campylobacter jejuni and 92 for campylobacter coli. positive samples originated from broilers, the broiler houses, cattle, pigs, bantams, laying hens, ...200818508146
flagellar glycosylation in clostridium botulinum.flagellins from clostridium botulinum were shown to be post-translationally modified with novel glycan moieties by top-down ms analysis of purified flagellin protein from strains of various toxin serotypes. detailed analyses of flagellin from two strains of c. botulinum demonstrated that the protein is modified by a novel glycan moiety of mass 417 da in o-linkage. bioinformatic analysis of available c. botulinum genomes identified a flagellar glycosylation island containing homologs of genes rec ...200818671733
genomic evidence for interspecies acquisition of chromosomal dna from campylobacter jejuni by campylobacter coli strains of a turkey-associated clonal group (cluster ii).previous multilocus sequence typing studies of campylobacter coli from meat animals identified an unusual cluster of strains, primarily from turkeys, termed "cluster ii" and characterized by the presence of the c. jejuni aspa103 allele. to characterize the extent of genomic input from c. jejuni in the aspa region of cluster ii c. coli, we sequenced the 6.1 kb genomic region upstream of and including aspa from two turkey-derived cluster ii strains (c. coli 6979 and c. coli 7474, of st-1150 and st ...200818673073
differential effects of temperature on natural transformation to erythromycin and nalidixic acid resistance in campylobacter coli.campylobacter jejuni and campylobacter coli are naturally competent, but limited information exists on the impact of environmental conditions on transformation. in this study, we investigated the impact of temperature and microaerobic versus aerobic atmosphere on transformation of c. coli to erythromycin and nalidixic acid resistance. frequency of transformation was not significantly different between microaerobic (5 to 10% co(2)) and aerobic conditions. however, c. coli was transformed to eryth ...200818708520
assessment of the stability of cell-surface components of microorganisms by maldi-tof-ms following preservation on lenticule discs.strains representing the species campylobacter coli, escherichia coli, listeria monocytogenes, pseudomonas aeruginosa, salmonella enterica, and staphylococcus aureus were randomly selected to assess the consistency of cells preserved on lenticule discs to those archived in traditional freeze-dried ampoules. each matched pair was cultured using identical conditions and analysed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (ms) to profile the surface-associated m ...200818717884
restoration of flagellar biosynthesis by varied mutational events in campylobacter jejuni.both a complex regulatory cascade involving the flgsr two-component system and phase variation control expression of sigma(54)-dependent flagellar genes in campylobacter jejuni. in this study, mutational mechanisms influencing production of the flgs histidine kinase were discovered. random non-motile, non-flagellated flgs variants were impaired for growth in the chick intestinal tract. spontaneous revertants restored for flagellar biosynthesis, gene expression, and motility identified by in vivo ...200818761684
identification of campylobacter jejuni proteins recognized by maternal antibodies of chickens.campylobacter jejuni is one of the leading bacterial causes of food-borne gastroenteritis. infection with c. jejuni is frequently acquired through the consumption of undercooked poultry or foods cross-contaminated with raw poultry. given the importance of poultry as a reservoir for campylobacter organisms, investigators have performed studies to understand the protective role of maternal antibodies in the ecology of campylobacter colonization of poultry. in a previous study, chicks with maternal ...200818805999
characterization of two campylobacter jejuni strains for use in volunteer experimental-infection studies.the development of vaccines against campylobacter jejuni would be facilitated by the ability to perform phase ii challenge studies. however, molecular mimicry of the lipooligosaccharide (los) of most c. jejuni strains with human gangliosides presents safety concerns about the development of guillain-barré syndrome. clinical isolates of c. jejuni that appeared to lack genes for the synthesis of ganglioside mimics were identified by dna probe analyses. two clinical isolates from southeast asia (st ...200818809665
[evaluation of antigenic properties of campylobacter jejuni and campylobacter coli proteins in a western-immunoblot].campylobacter jejuni and campylobacter coli are the most common bacterial cause for acute diarrheal illnesses in developed countries. the aim of this study was to evaluate the antigenic properties of campylobacterjejuni and campylobacter coli proteins in western-blot assay. whole-cell components of campulobacter jejuni and campylobacter coli were separated by sodium dodecyl sulfate-polyacrylamide gel electroforesis. using this method we detected in all seven c. jejuni strains 21 peptides migrati ...200818819448
development of a loop-mediated isothermal amplification assay for sensitive and rapid detection of vibrio parahaemolyticus.vibrio parahaemolyticus is a marine seafood-borne pathogen causing gastrointestinal disorders in humans. thermostable direct hemolysin (tdh) and tdh-related hemolysin (trh) are known as major virulence determinants of v. parahaemolyticus. most v. parahaemolyticus isolates from the environment do not produce tdh or trh. total v. parahaemolyticus has been used as an indicator for control of seafood contamination toward prevention of infection. detection of total v. parahaemolyticus using conventio ...200818823567
a temperature-regulated campylobacter jejuni gluconate dehydrogenase is involved in respiration-dependent energy conservation and chicken colonization.campylobacter jejuni is a gastrointestinal pathogen of humans but can asymptomatically colonize the avian gut. c. jejuni therefore grows at both 37 degrees c and 42 degrees c, the internal temperatures of humans and birds respectively. microarray and proteomic studies on temperature regulation in c. jejuni strain 81-176 revealed the upregulation at 42 degrees c of two proteins, cj0414 and cj0415, orthologous to gluconate dehydrogenase (gadh) from pectobacterium cypripedii. 81-176 demonstrated ga ...200818284594
longitudinal study of the molecular epidemiology of campylobacter jejuni in cattle on dairy farms.multilocus sequence typing (mlst), an accurate and phylogenetically robust characterization method for population studies of campylobacter, was applied to campylobacter jejuni isolates (n = 297) from the fecal samples of cattle from five dairy farms in cheshire, united kingdom, collected throughout 2003. the population dynamics of the c. jejuni strains, as identified by the occurrence of sequence types and clonal complexes, demonstrated variations within and between cattle populations over time. ...200818424539
identification of nine sequence types of the 16s rrna genes of campylobacter jejuni subsp. jejuni isolated from broilers.campylobacter is the most commonly reported bacterial cause of enteritis in humans in the eu member states and other industrialized countries. one significant source of infection is broilers and consumption of undercooked broiler meat. campylobacter jejuni is the campylobacter sp. predominantly found in infected humans and colonized broilers. sequence analysis of the 16s rrna gene is very useful for identification of bacteria to genus and species level. the objectives in this study were to deter ...200818492293
antimicrobial use through feed, water, and injection in 20 swine farms in alberta and saskatchewan.antimicrobial resistance (amr) is an emerging animal welfare and public health issue linked to antimicrobial use (amu) in livestock. this study was conducted in 2004 on 20 swine farms in alberta and saskatchewan. on-farm records and questionnaires were used to retrospectively describe the antimicrobial exposures of pigs through feed, water, and injection. antimicrobial use in all production categories was described over 12 months. on-farm records and questionnaires provided sufficient data to de ...200818505203
performance of a 70-mer oligonucleotide microarray for genotyping of campylobacter jejuni.campylobacter jejuni is widespread in the environment and is the major cause of bacterial gastroenteritis in humans. in the present study we use microarray-based comparative genomic hybridizations (cgh), pulsed-field gel electrophoresis (pfge) and multilocus sequence typing (mlst) to analyze closely related c. jejuni isolates from chicken and human infection.200818462507
detection of campylobacter jejuni and campylobacter coli from broiler chicken-related samples using bax pcr and conventional international organization for standardization culture.in this study, the conventional international organization for standardization (iso) culture method was compared with the dupont qualicon bax system, a high-throughput, rapid molecular assay that can be used to detect several bacterial species, including campylobacter jejuni and campylobacter coli in diverse sample types. standard enrichment culture is a time-consuming process, taking up to 6 days to obtain a confirmed result. rapid molecular assays have been developed that provide results withi ...200818468043
comparison of agar dilution and e-test for antimicrobial susceptibility testing of campylobacter coil isolates recovered from 80 ontario swine farms.the primary aim of this study was to evaluate the level of agreement of the e-test for in vitro antimicrobial susceptibility testing of campylobacter coli using the agar dilution technique, which is the approved method. a convenience sample of 80 ontario swine farms was chosen for this study; each farm was visited from january to june 2004. a total of 233 isolates of c. coli were tested for susceptibility to 10 antimicrobials by agar dilution and the e-test. performance of the tests was evaluate ...200818505206
the campylobacter jejuni thiol peroxidases tpx and bcp both contribute to aerotolerance and peroxide-mediated stress resistance but have distinct substrate specificities.the microaerophilic food-borne pathogen campylobacter jejuni experiences variable oxygen concentrations during its life cycle, especially during transitions between the external environment and the avian or mammalian gut. single knockout mutations in either one of two related thiol peroxidase genes, tpx and bcp, resulted in normal microaerobic growth (10% [vol/vol] oxygen) but poorer growth than that of the wild type under high-aeration conditions (21% [vol/vol] oxygen). however, a tpx/bcp doubl ...200818515414
development and evaluation of a loop-mediated isothermal amplification assay for rapid and simple detection of campylobacter jejuni and campylobacter coli.we developed a loop-mediated isothermal amplification (lamp) assay for the rapid and simple detection of campylobacter jejuni and campylobacter coli. the assay provides a specific lamp product for each of these two species. the assay correctly identified 65 c. jejuni and 45 c. coli strains, but not 75 non-c. jejuni/coli strains. the sensitivity of the lamp assay for c. jejuni and c. coli in spiked human stool specimens was 5.6 x 10(3) c.f.u. g(-1) (1.4 c.f.u. per test tube) and 4.8 x 10(3) c.f.u ...200818349363
viral, bacterial and parasitic etiology of pediatric diarrhea in gaza, palestine.to determine the etiology of acute diarrhea in palestinian children under 5 years of age and to improve knowledge of the etiology of gastrointestinal pathogens using traditional and molecular diagnostic techniques.200818523397
ccaat/enhancer binding protein beta is a major mediator of inflammation and viral replication in the gastrointestinal tract of simian immunodeficiency virus-infected rhesus macaques.the gastrointestinal tract (git) is a major target of infection with human immunodeficiency virus (hiv) and simian immunodeficiency virus (siv). chronic git disease and inflammation are common sequelae to hiv/siv infection. nonetheless, the molecular mechanisms that cause and maintain git dysfunction remain unclear. we investigated the contribution of ccaat/enhancer-binding protein beta (c/ebpbeta) to git disease and viral replication in jejunum and colon collected at necropsy from 12 siv-infect ...200818535173
key role of mfd in the development of fluoroquinolone resistance in campylobacter jejuni.campylobacter jejuni is a major food-borne pathogen and a common causative agent of human enterocolitis. fluoroquinolones are a key class of antibiotics prescribed for clinical treatment of enteric infections including campylobacteriosis, but fluoroquinolone-resistant campylobacter readily emerges under the antibiotic selection pressure. to understand the mechanisms involved in the development of fluoroquinolone-resistant campylobacter, we compared the gene expression profiles of c. jejuni in th ...200818535657
competing isogenic campylobacter strains exhibit variable population structures in vivo.consumption of poultry contaminated with campylobacter jejuni is a risk factor for human gastrointestinal disease. the rational development of control strategies for campylobacter within chickens requires an understanding of the colonization process at the molecular and population levels, both within and between hosts. experiments employing competing strains of campylobacter have been used to investigate colonization. implicit in these studies is the assumption that the behavior of competing str ...200818424530
prevalence and patterns of antimicrobial resistance of fecal escherichia coil among pigs on 47 farrow-to-finish farms with different in-feed medication policies in ontario and british columbia.the main objectives of this study were to determine the prevalence and patterns of antimicrobial resistance in pigs on farms that medicated swine ration and those that did not. a total of 940 isolates of escherichia coli from 188 pooled fecal samples obtained from weaner and finisher pigs on 47 farrow-to-finish swine farms (34 farms used in-feed medication and 13 did not) were tested for susceptibility to 21 antimicrobials using a breakpoint concentration method. the prevalence of resistance var ...200818505210
recombination-based in vivo expression technology identifies helicobacter pylori genes important for host colonization.here we undertook to identify colonization and gastric disease-promoting factors of the human gastric pathogen helicobacter pylori as genes that were induced in response to the stomach environment. using recombination-based in vivo expression technology (rivet), we identified six promoters induced in the host compared to laboratory conditions. three of these promoters, designated pivi10, pivi66, and pivi77, regulate genes that h. pylori may use to interact with other microbes or the host. pivi10 ...200818794279
helicobacter hepaticus hhgi1 is a pathogenicity island associated with typhlocolitis in b6.129-il10 tm1cgn mice.helicobacter hepaticus strain 3b1 (h. hepaticus) contains a genomic island of approximately 71 kb, hhgi1, with some of the common features shared among known bacterial pathogenicity islands. in this study, we characterized the pathogenic potential of hhgi1 by infecting b6.129-il10 tm1cgn (il10-/-) mice with an isogenic mutant (namely hhpaid1) lacking 19 predicted genes within hhgi1. in contrast to h. hepaticus (p<0.001), hhpaid1 did not cause typhlocolitis and hyperplasia in il10-/- mice. coloni ...200818538610
sensitive and rapid detection of cholera toxin-producing vibrio cholerae using a loop-mediated isothermal amplification.vibrio cholerae is widely acknowledged as one of the most important waterborne pathogen causing gastrointestinal disorders. cholera toxin (ct) is a major virulence determinant of v. cholerae. detection of ct-producing v. cholerae using conventional culture-, biochemical- and immunological-based assays is time-consuming and laborious, requiring more than three days. thus, we developed a novel and highly specific loop-mediated isothermal amplification (lamp) assay for the sensitive and rapid detec ...200818547441
expression, purification, and structural characterization of cfra, a putative iron transporter from campylobacter jejuni.the gene for the campylobacter ferric receptor (cfra), a putative iron-siderophore transporter in the enteric food-borne pathogen campylobacter jejuni, was cloned, and the membrane protein was expressed in escherichia coli, affinity purified, and then reconstituted into model lipid membranes. fourier transform infrared spectra recorded from the membrane-reconstituted cfra are similar to spectra that have been recorded from other iron-siderophore transporters and are highly characteristic of a be ...200818556796
comparative genomic analysis of clinical strains of campylobacter jejuni from south africa.campylobacter jejuni is a common cause of acute gastroenteritis and is also associated with the post-infectious neuropathies, guillain-barré and miller fisher syndromes. in the cape town area of south africa, c. jejuni strains with penner heat-stable (hs) serotype hs:41 have been observed to be overrepresented among cases of guillain-barré syndrome. the present study examined the genetic content of a collection of 32 south african c. jejuni strains with different serotypes, including 13 hs:41 st ...200818431496
influxed insects as vectors for campylobacter jejuni and campylobacter coli in danish broiler houses.the vector potential of flies (diptera: brachycera) for spread of campylobacter jejuni and campylobacter coli on 5 danish broiler farms was evaluated in a longitudinal field study from april to november 2004. first, the prevalence of c. jejuni- and c. coli-positive flies was determined in 2,816 flies captured from farm surroundings. each individual fly was macerated, preenriched in bolton broth for 24 h at 42 degrees c, streaked onto modified campylobater blood-free selective agar and incubated ...200818577626
effect of putative efflux pump inhibitors and inducers on the antimicrobial susceptibility of campylobacter jejuni and campylobacter coli.the cmeabc efflux pump plays an important role in the antimicrobial resistance of campylobacter jejuni and campylobacter coli. the aim of this investigation was to study the effect of putative efflux pump inhibitors, phenyl-arginine-beta-naphthylamide (pabetan) and 1-(1-naphthylmethyl)-piperazine (nmp), as well as the effect of putative efflux pump inducers, sodium salicylate and sodium deoxycholate, on the mic levels of erythromycin, ciprofloxacin, kanamycin, tetracycline and rifampicin for c. ...200818566143
comparative performance of different pcr assays for the identification of campylobacter jejuni and campylobacter coli.the sensitivity and specificity of 7 pcr assays described for the identification of campylobacter jejuni and campylobacter coli were examined using alkaline cell lysates from a collection of 100 well characterized reference strains of c. jejuni, c. coli, campylobacter lari and related campylobacter, helicobacter and arcobacter species. based on a preliminary evaluation, one multiplex test was excluded from further evaluation. the various assays differed considerably in sensitivity and specificit ...200818272350
high-resolution genotyping of campylobacter species by use of pcr and high-throughput mass spectrometry.in this work we report on a high-throughput mass spectrometry-based technique for the rapid high-resolution identification of campylobacter jejuni strain types. this method readily distinguishes c. jejuni from c. coli, has a resolving power comparable to that of multilocus sequence typing (mlst), is applicable to mixtures, and is highly automated. the strain typing approach is based on high-performance mass spectrometry, which "weighs" pcr amplicons with enough mass accuracy to unambiguously det ...200818272701
development of a strain-specific molecular method for quantitating individual campylobacter strains in mixed populations.the identification of sites resulting in cross-contamination of poultry flocks in the abattoir and determination of the survival and persistence of campylobacters at these sites are essential for the development of intervention strategies aimed at reducing the microbial burden on poultry at retail. a novel molecule-based method, using strain- and genus-specific oligonucleotide probes, was developed to detect and enumerate specific campylobacter strains in mixed populations. strain-specific oligo ...200818281428
development of a cytolethal distending toxin (cdt) gene-based species-specific multiplex pcr assay for the detection and identification of campylobacter jejuni, campylobacter coli and campylobacter fetus.a cytolethal distending toxin (cdt) gene-based species-specific multiplex pcr assay for the detection of cdta, cdtb or cdtc gene of campylobacter jejuni, campylobacter coli or campylobacter fetus, respectively, was developed and evaluated with 76 campylobacter strains belonging to seven different species and 131 other bacterial strains of eight different genera. the cdta, cdtb or cdtc gene of c. jejuni, c. coli or c. fetus, respectively, could be successfully amplified using the corresponding se ...200818248435
prevalence of gastrointestinal bacterial pathogens in a population of zoo animals.faecal prevalence of gastrointestinal bacterial pathogens, including campylobacter, escherichia coli o157:h7, salmonella, shigella, yersinia, as well as arcobacter, were examined in 317 faecal specimens from 44 animal species in belfast zoological gardens, during july-september 2006. thermophilic campylobacters including campylobacter jejuni, campylobacter coli and campylobacter lari, were the most frequently isolated pathogens, where members of this genus were isolated from 11 animal species (1 ...200818331520
differences in virulence attributes between cytolethal distending toxin positive and negative campylobacter jejuni strains.campylobacter jejuni is a common gastrointestinal bacterial pathogen. although cytolethal distending toxin (cdt) is proposed to be an important virulence determinant of this pathogen, how cdt(+) and cdt(-) strains differ in their biological properties remains largely unknown. the virulence properties of cdt(+) and cdt(-) strains were studied on hela cells and in the suckling mouse model. presence of the cdtb gene in campylobacter species was determined by pcr. five each of cdt(+) and cdt(-) c. j ...200818287287
prevalence of virulence genes and cytolethal distending toxin production in campylobacter jejuni isolates from diarrheal patients in bangladesh.from 300 stool samples, 58 campylobacter strains were isolated by standard microbiological and biochemical methods. of these, 40 strains were identified as campylobacter jejuni and 5 as campylobacter coli. the presence of flaa (100%), cadf (100%), racr (100%), dnaj (100%), plda (100%), ciab (95%), virb11 (0%), ceue (82.5%), cdta (97.5%), cdtb (97.5%), cdtc (97.5%), and wlan (7.5%) genes was detected in c. jejuni by pcr. all c. jejuni strains but one produced cytolethal distending toxin in a hela ...200818287317
antimicrobial resistance of campylobacter spp. isolated from broiler flocks in iceland 2001-2005.minimum inhibitory concentrations of six antimicrobial agents were determined for one campylobacter sp. isolate from each of the 362 campylobacter-positive commercial chicken flocks in iceland in the years 2001-2005. of all isolates tested, 6.9% were resistant, although none were multiresistant. resistance to ampicillin was most commonly observed (3.6%) followed by resistance to enrofloxacin (3%), nalidixic acid (1.9%), and oxytetracyclin (0.3%), with cross-resistance between enrofloxacin and na ...200818321203
evaluation of 16s rrna gene-based pcr assays for genus-level identification of helicobacter species.the inclusivity, exclusivity, and detection limit of six 16s rrna gene-based helicobacter genus-specific pcr assays were examined. five out of six assays were 100% inclusive, but the tests varied considerably in their exclusivity (9.1 to 95.5%). the clinical detection limit varied between 10(3) and 1 viable bacterial cell per reaction mixture.200818337392
sequence variability of campylobacter temperate bacteriophages.prophages integrated within the chromosomes of campylobacter jejuni isolates have been demonstrated very recently. prior work with campylobacter temperate bacteriophages, as well as evidence from prophages in other enteric bacteria, suggests these prophages might have a role in the biology and virulence of the organism. however, very little is known about the genetic variability of campylobacter prophages which, if present, could lead to differential phenotypes in isolates carrying the phages ve ...200818366706
comparison of antimicrobial resistance of campylobacter jejuni and campylobacter coli isolated from humans and chicken carcasses in poland.campylobacter-associated gastroenteritis remains an important cause of morbidity worldwide, and some evidence suggests that poultry is an important source of this foodborne infection in humans. this study was conducted to analyze the prevalence and genetic background of resistance of 149 campylobacter jejuni and 54 campylobacter coli strains isolated from broiler chicken carcasses and from stool samples of infected children in poland from 2003 through 2005. nearly all isolates were susceptible t ...200818389707
harmonised monitoring of antimicrobial resistance in salmonella and campylobacter isolates from food animals in the european union.many member states of the european union (eu) currently monitor antimicrobial resistance in zoonotic agents, including salmonella and campylobacter. according to directive 2003/99/ec, member states shall ensure that the monitoring provides comparable data on the occurrence of antimicrobial resistance. the european commission asked the european food safety authority to prepare detailed specifications for harmonised schemes for monitoring antimicrobial resistance. the objective of these specificat ...200818397331
convergence of campylobacter species: implications for bacterial evolution.the nature of species boundaries in bacteria remains controversial. in particular, the mechanisms of bacterial speciation and maintenance in the face of frequent genetic exchange are poorly understood. here, we report patterns of genetic exchange that show two closely related zoonotic pathogenic species, campylobacter jejuni and campylobacter coli, are converging as a consequence of recent changes in gene flow. population expansion into a novel ecological niche generated by human activity is the ...200818403712
comparison of campylobacter populations in wild geese with those in starlings and free-range poultry on the same farm.wild geese are a potential source of campylobacter infection for humans and farm animals and have been implicated in at least two large waterborne disease outbreaks. there have been few investigations into the population biology of campylobacter in geese, carriage rates are reported to vary (0 to 100%), and no genetic characterization of isolates has been performed. fecal samples collected from wild geese in oxfordshire, united kingdom, were culture positive for c. jejuni (50.2%) and c. coli (0. ...200818390684
campylobacter infection of broiler chickens in a free-range environment.campylobacter jejuni is the most common cause of bacterial gastroenteritis worldwide, with contaminated chicken meat considered to represent a major source of human infection. biosecurity measures can reduce c. jejuni shedding rates of housed chickens, but the increasing popularity of free-range and organic meat raises the question of whether the welfare benefits of extensive production are compatible with food safety. the widespread assumption that the free-range environment contaminates extens ...200818412548
risk factors for ciprofloxacin resistance in reported campylobacter infections in southern alberta.we conducted a case-control study examining risk factors for ciprofloxacin resistance in campylobacter infections that were reported in 2004 and 2005 in two health regions in southern alberta. the study questionnaire included questions about recent travel and antibiotic use, food consumption frequency, use of household and personal hygiene products with antibacterial agents, contact with animals, and potential misuse of antibiotics. of the 210 patients who participated, 31.0% had ciprofloxacin-r ...200817678560
risk factors for ciprofloxacin resistance in reported campylobacter infections in southern alberta.we conducted a case-control study examining risk factors for ciprofloxacin resistance in campylobacter infections that were reported in 2004 and 2005 in two health regions in southern alberta. the study questionnaire included questions about recent travel and antibiotic use, food consumption frequency, use of household and personal hygiene products with antibacterial agents, contact with animals, and potential misuse of antibiotics. of the 210 patients who participated, 31.0% had ciprofloxacin-r ...200817678560
wide geographical distribution of internationally rare campylobacter clones within new zealand.during the southern hemisphere winter of 2006 new zealand experienced a significant increase in the number of reported cases of campylobacter infection. in total, 112 campylobacter isolates from eight district health boards (dhbs) located across new zealand were submitted for pfge, mlst and penner serotyping analysis. distinct clusters of campylobacter isolates were identified, several of which were composed of isolates from up to five different dhbs located on both the north and south islands o ...200818028582
wide geographical distribution of internationally rare campylobacter clones within new zealand.during the southern hemisphere winter of 2006 new zealand experienced a significant increase in the number of reported cases of campylobacter infection. in total, 112 campylobacter isolates from eight district health boards (dhbs) located across new zealand were submitted for pfge, mlst and penner serotyping analysis. distinct clusters of campylobacter isolates were identified, several of which were composed of isolates from up to five different dhbs located on both the north and south islands o ...200818028582
production of a monoclonal antibody specific for the major outer membrane protein of campylobacter jejuni and characterization of the epitope.campylobacter species are important enteric pathogens causing disease in humans and animals. there is a lack of a good immunological test that can be used routinely to separate campylobacter jejuni from other campylobacter species. we produced monoclonal antibodies (mabs) directed against the major outer membrane protein (momp) of c. jejuni using recombinant momp as the antigen. one mab, designated mab5c4 and of the immunoglobulin g1 isotype, was found to be potentially specific for c. jejuni. d ...200818065632
production of a monoclonal antibody specific for the major outer membrane protein of campylobacter jejuni and characterization of the epitope.campylobacter species are important enteric pathogens causing disease in humans and animals. there is a lack of a good immunological test that can be used routinely to separate campylobacter jejuni from other campylobacter species. we produced monoclonal antibodies (mabs) directed against the major outer membrane protein (momp) of c. jejuni using recombinant momp as the antigen. one mab, designated mab5c4 and of the immunoglobulin g1 isotype, was found to be potentially specific for c. jejuni. d ...200818065632
survey of campylobacter jejuni in stray cats in southern italy.to evaluate the presence of thermotolerant campylobacter spp. in stray cats in southern italy.200818069980
lack of response of int-407 cells to the presence of non-culturable campylobacter jejuni.many contradictory articles on the infectivity of non-culturable campylobacter jejuni can be found. we studied the effect of non-culturable c. jejuni in an in vitro assay. to prevent the potential effect of a few culturable bacteria in the non-culturable suspension, int-407 cells, which mimic the outer cell layer in the small intestines, were exposed to culturable c. jejuni suspensions with or without non-culturable c. jejuni. the number of bacteria adhering to and/or invading int-407 cells and ...200818081950
lack of response of int-407 cells to the presence of non-culturable campylobacter jejuni.many contradictory articles on the infectivity of non-culturable campylobacter jejuni can be found. we studied the effect of non-culturable c. jejuni in an in vitro assay. to prevent the potential effect of a few culturable bacteria in the non-culturable suspension, int-407 cells, which mimic the outer cell layer in the small intestines, were exposed to culturable c. jejuni suspensions with or without non-culturable c. jejuni. the number of bacteria adhering to and/or invading int-407 cells and ...200818081950
fla-dgge analysis of campylobacter jejuni and campylobacter coli in cecal samples of broilers without cultivation.in a commercial broiler flock during rearing multiple genotypes of campylobacter jejuni may be present as well as in gastrointestinal tracts of individual birds. the aim of this study was to optimize and apply a denaturing gradient gel electrophoresis assay of the flagellin gene (fla-dgge) for analysis of c. jejuni and campylobacter coli in cecal samples of broilers without prior cultivation. one c. coli and 21 c. jejuni strains isolated from broiler flocks, of which 14 typed as unique by restri ...200817890022
sexual dimorphism in campylobacteriosis.sexual dimorphism in infectious diseases whereby disease incidence is more prevalent in one gender has been reported repeatedly in the scientific literature. both behavioural and physiological differences have been suggested as a cause of this gender bias but there is a paucity of data to support either of these viewpoints. here it is hypothesized that for campylobacteriosis physiological factors play an important role in the higher incidence in males. we demonstrate in the human population (fro ...200818062834
sexual dimorphism in campylobacteriosis.sexual dimorphism in infectious diseases whereby disease incidence is more prevalent in one gender has been reported repeatedly in the scientific literature. both behavioural and physiological differences have been suggested as a cause of this gender bias but there is a paucity of data to support either of these viewpoints. here it is hypothesized that for campylobacteriosis physiological factors play an important role in the higher incidence in males. we demonstrate in the human population (fro ...200818062834
characterization of two putative cytochrome c peroxidases of campylobacter jejuni involved in promoting commensal colonization of poultry.campylobacter jejuni is a leading cause of bacterial gastroenteritis in humans throughout the world, but infection of animals, especially poultry, results in a commensal colonization of the intestines. we previously found that a mutant lacking doca, which encodes a putative cytochrome c peroxidase (ccp), demonstrates up to a 10(5)-fold reduction in colonization of the chick cecum compared to wild-type c. jejuni strain 81-176. predictions from genomic sequences identified cjj0382 as a second locu ...200818086814
characterization of two putative cytochrome c peroxidases of campylobacter jejuni involved in promoting commensal colonization of poultry.campylobacter jejuni is a leading cause of bacterial gastroenteritis in humans throughout the world, but infection of animals, especially poultry, results in a commensal colonization of the intestines. we previously found that a mutant lacking doca, which encodes a putative cytochrome c peroxidase (ccp), demonstrates up to a 10(5)-fold reduction in colonization of the chick cecum compared to wild-type c. jejuni strain 81-176. predictions from genomic sequences identified cjj0382 as a second locu ...200818086814
role of campylobacter jejuni respiratory oxidases and reductases in host colonization.campylobacter jejuni is the leading cause of human food-borne bacterial gastroenteritis. the c. jejuni genome sequence predicts a branched electron transport chain capable of utilizing multiple electron acceptors. mutants were constructed by disrupting the coding regions of the respiratory enzymes nitrate reductase (napa::cm), nitrite reductase (nrfa::cm), dimethyl sulfoxide, and trimethylamine n-oxide reductase (termed cj0264::cm) and the two terminal oxidases, a cyanide-insensitive oxidase (cy ...200818192421
retrospective study of campylobacter infection in a zoological collection.little is known about the epidemiology of campylobacter spp. in wild animal populations. however, zoological collections can provide valuable insights. using records from the zoological society of london whipsnade zoo compiled between 1990 and 2003, the roles of a range of biotic and abiotic factors associated with the occurrence of campylobacteriosis were investigated. the occurrence of campylobacteriosis varied widely across host taxonomic orders. furthermore, in mammals, a combination of chan ...200818165368
retrospective study of campylobacter infection in a zoological collection.little is known about the epidemiology of campylobacter spp. in wild animal populations. however, zoological collections can provide valuable insights. using records from the zoological society of london whipsnade zoo compiled between 1990 and 2003, the roles of a range of biotic and abiotic factors associated with the occurrence of campylobacteriosis were investigated. the occurrence of campylobacteriosis varied widely across host taxonomic orders. furthermore, in mammals, a combination of chan ...200818165368
real-time pcr approach for detection of environmental sources of campylobacter strains colonizing broiler flocks.reducing colonization of poultry flocks by campylobacter spp. is a key strategy in the control and prevention human campylobacteriosis. horizontal transmission of campylobacters, from in and around the farm, is the presumed route of flock colonization. however, the identification and prioritization of sources are confounded by the ubiquitous nature of these organisms in the environment, their poor rates of recovery by standard culture methods, and the need for cost-effective and timely methods f ...200818203857
risk factors for infection with campylobacter jejuni flaa genotypes.we aimed to explore campylobacter genotype-specific risk factors in australia. isolates collected prospectively from cases recruited into a case-control study were genotyped using flaa restriction fragment-length polymorphism typing (flaa genotyping). exposure information for cases and controls was collected by telephone interview. risk factors were examined for major flaa genotypes using logistic and multinomial regression. five flaa genotypes accounted for 325 of 590 (55%) cases - flaa-6b (n=1 ...200818205975
campylobacter jejuni survives within epithelial cells by avoiding delivery to lysosomes.campylobacter jejuni is one of the major causes of infectious diarrhea world-wide, although relatively little is know about its mechanisms of pathogenicity. this bacterium can gain entry into intestinal epithelial cells, which is thought to be important for its ability to persistently infect and cause disease. we found that c. jejuni is able to survive within intestinal epithelial cells. however, recovery of intracellular bacteria required pre-culturing under oxygen-limiting conditions, suggesti ...200818225954
the origin of a novel gene through overprinting in escherichia coli.overlapped genes originate by a) loss of a stop codon among contiguous genes coded in different frames; b) shift to an upstream initiation codon of one of the contiguous genes; or c) by overprinting, whereby a novel open reading frame originates through point mutation inside an existing gene. although overlapped genes are common in viruses, it is not clear whether overprinting has led to new genes in prokaryotes.200818226237
functional characterization and mutagenesis of the proposed behavioral sensor tlpd of helicobacter pylori.helicobacter pylori requires flagellar motility and chemotaxis to establish and maintain chronic infection of the human stomach. the ph gradient in the stomach mucus is essential for bacterial orientation and guides the bacterium toward a narrow layer of the mucus, suggesting that h. pylori is capable of energy sensing or taxis. in the present study, h. pylori wild-type behavior in a temporal swimming assay could be altered by electron transport inhibitors, indicating that a connection between m ...200818245281
case-case analysis of enteric diseases with routine surveillance data: potential use and example results.case-control studies and outbreak investigations are the major epidemiological tools for providing detailed information on enteric disease sources and risk factors, but these investigations can be constrained by cost and logistics.200818976484
dna mimics for the rapid identification of microorganisms by fluorescence in situ hybridization (fish).fluorescence in situ hybridization (fish) is a well-established technique that is used for a variety of purposes, ranging from pathogen detection in clinical diagnostics to the determination of chromosomal stability in stem cell research. the key step of fish involves the detection of a nucleic acid region and as such, dna molecules have typically been used to probe for the sequences of interest. however, since the turn of the century, an increasing number of laboratories have started to move on ...200819325728
occurrence and characterization of campylobacter in the brazilian production and processing of broilers.twenty-two commercial broiler flocks and their carcasses, totaling 546 samples (450 collected from a poultry farm and 96 from a slaughterhouse), were surveyed for the presence of campylobacter. the positive results for campylobacter among the analyzed samples were homogeneous, yielding 81.8% for cecal droppings, 80.9% for feces, and 80.4% for cloacal swabs. pre-enrichment and direct plating showed that 77.85% and 81.8% of cloacal swabs, respectively, were positive for campylobacter compared to 9 ...200819166063
evidence of cross-contamination by campylobacter spp. of broiler carcasses using genetic characterization of isolates.campylobacter is recognized as one of the leading cause of gastroenteritis worldwide, and is frequently isolated from the small intestines and ceca microflora of chickens. twenty-one out of 81 campylobacter-positive poultry flocks were selected to evaluate the genetic diversity of campylobacter isolates and to study the distribution of genotypes among flocks. campylobacter isolates recovered from chicken carcasses and ceca were analyzed by pulsed-field gel electrophoresis (pfge). little diversit ...200819086371
antimicrobial resistance in selected bacteria from poultry.this paper reviews the present state of antimicrobial resistance (amr) in the zoonotic bacteria salmonella, campylobacter jejuni and campylobacter coli, and in escherichia coli from chickens and turkeys. for salmonella, the frequencies and patterns of amr vary depending on time, region, serovar, the particular farm, layers versus broilers, and the antimicrobial agent. there is usually a higher frequency of amr in salmonella from turkeys compared with salmonella from chickens. clonal and horizont ...200819102788
[annual changes in antimicrobial susceptibility of campylobacter jejuni and campylobacter coli isolated from acute diarrheic patients].two hundred thirty-one campylobacter were isolated from acute diarrheic patients between january 2001 and december 2005. we evaluated annual changes in identified species of campylobacter and their susceptibilities against antibiotics. campylobacter jejuni (219 strains; 94.8%) and campylobacter coli (12 strains; 5.2%) were identified to the species. susceptibilities to four antimicrobial agents, minocycline (mino), levofloxacin (lvfx), erythromycin (em) and clindamycin (cldm) were examined. the ...200819086420
trends of norfloxacin and erythromycin resistance of campylobacter jejuni/campylobacter coli isolates recovered from international travelers, 1994 to 2006.campylobacter sp. is a major cause of bacterial enterocolitis and travelers' diarrhea. empiric treatment regimens include fluoroquinolones and macrolides.200819090796
multilocus sequence typing, pulsed-field gel electrophoresis, and fla short variable region typing of clonal complexes of campylobacter jejuni strains of human, bovine, and poultry origins in luxembourg.campylobacter jejuni is the most common cause of bacterial gastroenteritis in luxembourg, with a marked seasonal peak during summer. the majority of these infections are thought to be sporadic, and the relative contribution of potential sources and reservoirs is still poorly understood. we monitored human cases from june to september 2006 (n = 124) by molecular characterization of isolates with the aim of rapidly detecting temporally related cases. in addition, isolates from poultry meat (n = 36 ...200818931296
genes and molecules of lactobacilli supporting probiotic action.lactobacilli have been crucial for the production of fermented products for centuries. they are also members of the mutualistic microbiota present in the human gastrointestinal and urogenital tract. recently, increasing attention has been given to their probiotic, health-promoting capacities. many human intervention studies demonstrating health effects have been published. however, as not all studies resulted in positive outcomes, scientific interest arose regarding the precise mechanisms of act ...200819052326
helicobacter pylori hopq outer membrane protein attenuates bacterial adherence to gastric epithelial cells.helicobacter pylori genomes contain about 30 hop genes that encode outer membrane proteins. helicobacter pylori hopq alleles exhibit a high level of genetic diversity, and two families of hopq alleles have been described. type i hopq alleles are found more commonly in cag-positive h. pylori strains from patients with peptic ulcer disease than in cag-negative strains from patients without ulcer disease. in this study, we mutated hopq in four h. pylori strains that each contained a type i hopq all ...200819065710
[evaluation of usefulness of commercial recomwell campylobacter enzyme--linked immunosorbent assays for routine serodiagnosis of campylobacter jejuni and campylobacter coli infections].the commercially available enzyme-linked immunosorbent assays (elisa recomwell campylobacter) from mikrogen was evaluated for the diagnosis of campylobacter jejuni and campylobacter coli infections. serum samples from 20 healthy controls, 44 persons with symptoms of primary campylobacter infection and 24 serum samples from patients with yersinia enterocolitica or salmonella infections were tested. this elisa assay detects iga and igg antibodies against three recombinant antigens of the campyloba ...200819382600
campylobacter enteritis in children in northern taiwan--a 7-year experience.campylobacter infection is recognized as a major cause of acute enteritis in humans. the clinical characteristics may vary in different countries. this study investigated the clinical manifestations of pediatric campylobacter enteritis in a medical center in northern taiwan.200819122923
assessing sensitivity to antibacterial topoisomerase ii inhibitors.both prokaryotes and eukaryotes have two major classes of topoisomerases that make transient single- or double-strand cuts in dna. while these enzymes play critical roles in cellular processes, they are also important targets of therapeutic agents. this unit describes assays to use in characterizing topoisomerase ii-targeting agents in vitro and in bacterial cells. it provides protocols for characterizing the action of small molecules against bacterial type ii topoisomerases in vitro and the in ...200721948169
the flik protein and flagellar hook-length control.the bacterial flagellum is a highly complex prokaryotic organelle. it is the motor that drives bacterial motility, and despite the large amount of energy required to make and operate flagella, motile organisms have a strong adaptive advantage. flagellar biogenesis is both complex and highly coordinated and it typically involves at least three two-component systems. part of the flagellum is a type iii secretion system, and it is via this structure that flagellar components are exported. the assem ...200717456739
antibiotic resistance and resistance mechanisms in campylobacter jejuni and campylobacter coli.campylobacter jejuni and campylobacter coli are recognized as the most common causative agents of bacterial gastroenteritis in the world and infections with these organisms occur more frequently than do infections due to salmonella species, shigella species, or escherichia coli 0157:h7. the incidence of human campylobacter infections has increased markedly in both developed and developing countries worldwide and, more significantly, so has the rapid emergence of antibiotic-resistant campylobacte ...200718031331
characterization of campylobacter phages including analysis of host range by selected campylobacter penner serotypes.the predominant food borne pathogen in the western world today is campylobacter. campylobacter specific bacteriophages (phages) have been proposed as an alternative agent for reducing the burden of campylobacter in broilers. one concern in relation to phage biocontrol is the narrow host range often displayed by phages. to identify the potential of phages as a campylobacter reducing agent we needed to determine their infectivity on a panel of isolates representing the campylobacter strains found ...200717945022
pulsed-plasma gas-discharge inactivation of microbial pathogens in chilled poultry wash water.a pulsed-plasma gas-discharge (ppgd) system was developed for the novel decontamination of chilled poultry wash water. treatment of poultry wash water in the plasma generation chamber for up to 24 s at 4 degrees c reduced escherichia coli nctc 9001, campylobacter jejuni atcc 33560, campylobacter coli atcc 33559, listeria monocytogenes nctc 9863, salmonella enterica serovar enteritidis atcc 4931, and s. enterica serovar typhimurium atcc 14028 populations to non-detectable levels (< or = 8 log cfu ...200718095434
the complete genome sequence and analysis of the epsilonproteobacterium arcobacter butzleri.arcobacter butzleri is a member of the epsilon subdivision of the proteobacteria and a close taxonomic relative of established pathogens, such as campylobacter jejuni and helicobacter pylori. here we present the complete genome sequence of the human clinical isolate, a. butzleri strain rm4018.200718159241
role of poultry meat in sporadic campylobacter infections in bosnia and herzegovina: laboratory-based study.to investigate genetic diversity and specificity of campylobacter jejuni and campylobacter coli strains isolated from humans, retail poultry meat, and live farm chickens in zenica-doboj canton, bosnia and herzegovina, and identify the role of poultry meat in sporadic campylobacter infections.200718074419
gastrointestinal disease in simian immunodeficiency virus-infected rhesus macaques is characterized by proinflammatory dysregulation of the interleukin-6-janus kinase/signal transducer and activator of transcription3 pathway.gastrointestinal disease and inflammation are common sequelae of human and simian immunodeficiency virus (siv) infection. nevertheless, the molecular mechanisms that lead to gastrointestinal dysfunction remain unclear. we investigated regulation of the interleukin (il)-6-jak-stat3 pathway in jejunum and colon, collected at necropsy, from 10 siv-infected macaques with diarrhea (group 1), 10 non-siv-infected macaques with diarrhea (group 2), and 7 control uninfected macaques (group 3). all group 1 ...200718055558
cryptic plasmids isolated from campylobacter strains represent multiple, novel incompatibility groups.three small, cryptic plasmids from the multi-drug-resistant (mdr) campylobacter coli strain rm2228 and one small, cryptic plasmid from the mdr campylobacter jejuni strain rm1170 were sequenced and characterized. pcc2228-1 has some similarity to firmicutes repl family plasmids that replicate via a rolling-circle mechanism. pcc2228-2 is a theta-replicating, iteron-containing plasmid (icp) that is a member of the same incompatibility (inc) group as previously described campylobacter shuttle vectors ...200717064774
longitudinal study of finnish campylobacter jejuni and c. coli isolates from humans, using multilocus sequence typing, including comparison with epidemiological data and isolates from poultry and cattle.we describe a study on the application of multilocus sequence typing for the analysis of campylobacter jejuni and c. coli isolates from human domestically acquired infections in the helsinki-uusimaa area of finland in 1996, 2002, and 2003. in addition, isolates from poultry meat and fecal samples of cattle from the seasonal peak (july to september) in 2003 were included in the study. in total, 361 finnish c. jejuni and c. coli strains were typed. sequence type 45 (st-45) (45%), st-21 (21%), and ...200717085689
prevalence of campylobacter jejuni and campylobacter coli in chicken hybrids with different growth rates, reared according to conventional and "free-range" production methods. 200717682919
(e)-methylisoeugenol and elemicin: antibacterial components of daucus carota l. essential oil against campylobacter jejuni.the essential oil of wild daucus carota l. obtained from aerial parts at the end of the flowering stage (dceo) was reported as antimicrobial against the human enteropathogen campylobacter jejuni. the aim of the present study was to extend this analysis to other campylobacter species and to identify the active compounds of the essential oil, subjected to gc, gc-ms, and (13)c nmr analysis. a minimum inhibitory concentration assay was used to quantify the antimicrobial activity of dceo and the majo ...200717685629
crystal structure of the transcriptional regulator cmer from campylobacter jejuni.the cmeabc multidrug efflux pump, which belongs to the resistance-nodulation-division (rnd) family, recognizes and extrudes a broad range of antimicrobial agents and is essential for campylobacter jejuni colonization of the animal intestinal tract by mediating the efflux of bile acids. the expression of cmeabc is controlled by the transcriptional regulator cmer, whose open reading frame is located immediately upstream of the cmeabc operon. to understand the structural basis of cmer regulation, w ...200717686491
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