J Appl Microbiol, 2004, 96(4), 795 - 802 Genetic relatedness among Campylobacter jejuni serotyped isolates of diverse origin as determined by numerical analysis of amplified fragment length polymorphism (AFLP) profiles; Siemer BL et al.; AIMS: To use amplified fragment length polymorphism (AFLP) analysis to evaluate the genetic relatedness among 254 Campylobacter jejuni reference and field strains of diverse origin representing all defined 'Penner' serotypes for this species . METHODS AND RESULTS: Field strains (n = 207) from human diarrhoea and diverse animal and environmental sources were collected mainly through a National surveillance programme in Denmark and serotyped by use of the established 'Penner' scheme . Genetic relationships among these isolates, and the archetypal serotype reference strains, were assessed by numerical analysis of AFLP profiles derived from genomic DNA . Extensive genetic diversity was seen among the strains examined; however, 43 groups of isolates were identified at the 92% similarity (S-) level . Thirteen groups contained isolates from a single host, possibly representing genotypes of 'low risk' to human health . The remaining 30 groups contained isolates from humans, chickens and associated food products, cattle, sheep, turkeys, ostriches and/or dogs . Strains assigned to serotypes 2, 6/7, 11 and 12 formed major clusters at the 77.6% S-level . Most other serotypes did not form homogeneous clusters . CONCLUSIONS: High-resolution genotyping applied to strains from a comprehensive range of sources provides evidence for multiple sources of sporadic C . jejuni infection . The results suggest that public health protection measures should be directed at all foods of animal origin . SIGNIFICANCE AND IMPACT OF THE STUDY: The genetic relatedness among all 'Penner' serotypes of C . jejuni is assessed by AFLP analysis . In addition, further evidence of epidemic and host-specific clones of C . jejuni is provided. J Appl Microbiol, 2004, 96(4), 664 - 70 Enhanced recovery and isolation of Campylobacter spp . from water using a novel device; Baserisalehi M et al.; AIM: To design a special device which can be used with nonselective blood-free nutrient agar without enrichment for detection of campylobacters from water . METHODS AND RESULTS: The Kapadnis-Baseri device (KB device) was designed and evaluated in comparison with the conventional method (C method) for detection of Campylobacter spp . from river water samples . The results indicated that the recovery of Campylobacter spp . by KB device was relatively more than by C method . CONCLUSIONS: To date, the methods for recovery of campylobacters are time consuming and involve use of special culture media, which is cost ineffective . The KB device is designed based on two important characters of Campylobacter, viz . motility and activity at low temperature . With this device we isolated Campylobacter spp . from river water on nonselective media without enrichment . Thus this device is as effective as the use of antibiotic media for the isolation of campylobacters . SIGNIFICANCE AND IMPACT OF THE STUDY: The KB device will be useful for isolation of Campylobacter spp . from water and other environmental samples, which is less time consuming and inexpensive . Besides, this device allows isolation of antibiotic sensitive campylobacters. Neurology, 2004 Mar 9, 62(5), 825 - 7 Acute facial diplegia and hyperreflexia: A Guillain-Barré syndrome variant; Susuki K et al.; Two patients with acute facial diplegia and hyperreflexia are described . Both patients had serologic evidence of preceding Campylobacter jejuni infection and antiganglioside IgG antibodies as well as other laboratory and electrophysiologic findings suggesting Guillain-Barre syndrome (GBS) . IV immunoglobulin produced recovery . Hyperreflexia does not necessarily exclude the diagnosis of a GBS variant . Antiganglioside antibodies can help with diagnosis in difficult cases. Rev Sci Tech, 2003 Dec, 22(3), 1013 - 20 {Campylobacteriosis in a low-income community in Buenos Aires, Argentina}; Lopez C et al.; The authors describe the problem of campylobacteriosis in a low-income community located in southern Buenos Aires . Homes in the area were classified according to their total number of inhabitants, the number of children and animals living in the house and the amount of poultry meat consumed . Samples were taken from all the different types of homes that had been identified in order to isolate, identify and type Campylobacter sp., using the modified Skirrow technique, bio-chemical tests and the Lior technique respectively . The prevalences found were 16.96% in dogs, 20% in cats and 40% in poultry . The pathogens isolated were Campylobacter jejuni type II in 94.44% of the cases and Campylobacter upsaliensis in 5.55% of the cases . In poultry meat, 92.85% of the samples were positive (23.07% for type I and 76.92% for type II) . The incidence of diarrhoea in children caused by C . jejuni was 0.4/1000 children/month (type I in all cases) . A high prevalence of C . jejuni type II was detected in pets and in poultry meat . The source of infection in children may have been poultry meat. J Clin Microbiol, 2004 Mar, 42(3), 1363 - 4 Occurrence of Campylobacter jejuni in pets living with human patients infected with C . jejuni; Damborg P et al.; 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. J Clin Microbiol, 2004 Mar, 42(3), 1313 - 5 Correlation between detection rates of periodontopathic bacterial DNA in carotid coronary stenotic artery plaque and in dental plaque samples; Ishihara K et al.; Utilizing PCR, the 16S rRNA detection rates for Porphyromonas gingivalis, Actinobacillus actinomycetemcomitans, Bacteroides forsythus, Treponema denticola, and Campylobacter rectus in samples of stenotic coronary artery plaques were determined to be 21.6, 23.3, 5.9, 23.5, and 15.7%, respectively . The detection rates for P . gingivalis and C . rectus correlated with their presence in subgingival plaque. Plasmid, 2004 Mar, 51(2), 101 - 7 Construction of replicative and integrative plasmids for setting up the in vivo expression technology in Helicobacter pylori; Angelini F et al.; Some pathogenic factors of Helicobacter pylori, a bacterium involved in peptic ulcer and gastric cancer, have already been identified using either global or particular approach, but there are still some orphan genes and unidentified pathogenic factors . One of the methods used successfully for the identification of virulence genes of many pathogens is the in vivo expression technology . We describe here the construction and sequences of three different plasmids, one integrative and two replicatives, for the identification of virulence genes by using in vivo expression technology in H . pylori, and of potential use in other bacteria such as Campylobacter spp . Moreover, the use of the green fluorescent protein could allow to classify the genes according to the strength of their expression and to identify those which are repressed upon interaction with gastric mucosa. J Infect, 2004 Apr, 48(3), 236 - 44 Induction of alpha and beta chemokines by intestinal epithelial cells stimulated with Campylobacter jejuni; Bakhiet M et al.; OBJECTIVES: To investigate the production of dynamic alpha and beta chemokines represented by interleukin-8 (IL-8) as alpha chemokine and CCL2 (monocyte-chemoattractant protein-1, CCR2 ligand), CCL4 (macrophage-inflammatory protein-1beta, CCR5 ligand), CCL3 (macrophage-inflammatory protein-1alpha, CCR1/5 ligand), (CCL5, regulated upon activation, normal T-cell expressed and secreted (RANTES, CCR5 ligand) as beta chemokines by the human intestinal cell line INT407 stimulated with factors produced by living Campylobacter jejuni (C . jejuni) and those present within sonicated and filtrated bacteria . METHODS: We used immunohistochemical technique modified to detect intracellular production of cytokines protein and RT-PCR to read RNA messages for evaluation of de novo cytokine synthesis . RESULTS: Living bacteria induced increased numbers of IL-8, CCL4 and CCL2 but not CCL3 or CCL5 producing cells . Low numbers of IL-8, CCL4 and CCL2 producing cells were detected with filtrated supernatant compared to living and sonicated bacteria . A non-significant low number of chemokine producing cells was noted when comparing numbers of chemokine producing cells stimulated with living C . jejuni to those stimulated with sonicated bacteria, indicating that the triggering factors involved in stimulation with living bacteria were still active after sonication, but they were largely lost upon filtration . The mRNA signals for IL-8 were noted in conformity with its protein levels as increased IL-8 mRNA signals were registered after stimulation with living and sonicated bacteria but not with filtrated supernatant . CONCLUSIONS: Preferential production of chemokines probably induced by membrane associate factors of C . jejuni acting on intestinal epithelial cells is presented . These chemokines are suggested to be part of an inflammatory network affecting cell types that contribute to initiation and/or resolution of the infection. Microb Drug Resist, 2003 Winter, 9(4), 373 - 9 Public health consequences of use of antimicrobial agents in food animals in the United States; Anderson AD et al.; The use of antimicrobial agents in food animals has caused concern regarding the impact these uses have on human health . Use of antimicrobial agents in animals and humans results in the emergence and dissemination of resistant bacteria . Resistant bacteria from food animals may be passed through the food chain to humans resulting in resistant infections . Increasing resistance to antimicrobial agents that are important in the treatment of human diseases, such as fluoroquinolones and third-generation cephalosporins for the treatment of Salmonella and Campylobacter infections, has significant public health implications . Efforts to mitigate the effects of increasing resistance require collaboration by several partners, including the farming, veterinary, medical, and public health communities. J Rheumatol, 2004 Mar, 31(3), 528 - 30 Reactive arthritis following an outbreak of Campylobacter jejuni infection; Hannu T et al.; OBJECTIVE: To study the occurrence and the clinical picture of musculoskeletal (MSK) complications including reactive arthritis (ReA) following an outbreak of Campylobacter jejuni . METHODS: An outbreak of C . jejuni infection occurred in 2000 in Asikkala, Finland, during which 350 exposed subjects contacted the Municipal Health Centre (MHC) . All primary care physicians in the MHC were advised to refer patients with acute MSK complications to the Rheumatism Foundation Hospital (RFH) for a specialist clinical examination, which was performed <or= 3 months after the onset of the outbreak . RESULTS: Fifteen subjects with acute MSK complaints (11 women, 4 men; mean age 58 yrs) were examined in the RFH, where the following MSK diagnoses were assessed: ReA (9 patients), reactive arthralgia (2) . exacerbation of previous rheumatoid arthritis (3) . and previous fibromyalgia (1) . In the patients with ReA, all adults, the arthritis was oligoarticular in 6 patients and polyarticular in 3; one patient had monoarthritis . The most frequently affected joints were knees and ankles . Besides peripheral arthritis, one patient had clinical sacroiliitis . Of the ReA patients, the antigen HLA-B27 was positive in 33%, including the patient with sacroiliitis . At the clinical examination, 6 ReA patients had subsiding signs of synovitis, 2 had only arthralgia, and one was symptom-free . CONCLUSION: The frequency of ReA following an outbreak of C . jejuni was low: 2.6% (9 of 350) . In the ReA patients, the clinical picture was mild, the primary outcome good, and the association with HLA-B27 not high. Microbiology, 2004 Mar, 150(Pt 3), 561 - 9 Campylobacter jejuni infection of differentiated THP-1 macrophages results in interleukin 1 beta release and caspase-1-independent apoptosis; Siegesmund AM et al.; Apoptosis induction of host macrophages has emerged as a common virulence mechanism among bacterial pathogens . Infection with Campylobacter jejuni is a leading cause of gastroenteritis worldwide and is characterized by an acute inflammatory response in the small intestine . The authors used the human monocytic cell line THP-1 to examine apoptosis induction and pro-inflammatory cytokine production during C . jejuni infection . Flow cytometric analysis revealed that 48 h after inoculation, a C . jejuni wild-type isolate induced apoptosis in 63 % of THP-1 cells while only 34 % of cells inoculated with a ciaB mutant, which does not secrete the Cia (Campylobacter invasion antigens) proteins, underwent apoptosis . Complementation of the ciaB mutant resulted in levels of apoptosis similar to those induced by the C . jejuni wild-type isolate, suggesting that the Cia proteins have a role in apoptosis induction . It was shown that a proteinase K- and heat-stable component of C . jejuni also stimulated THP-1 apoptosis . Inoculation with a C . jejuni gmhD mutant indicated that lipooligosaccharide was not the stimulatory molecule . Immunoblot and ELISA analyses revealed that C . jejuni infection stimulated the synthesis, processing and secretion of interleukin 1 beta (IL-1 beta) . Inhibition of caspase 1 activity eliminated IL-1 beta processing and secretion, but did not affect apoptosis induction . In addition, treatment of cells with a caspase-9-specific inhibitor did not affect apoptosis induction, arguing against activation of an apoptotic pathway dependent on either caspase 1 or 9 activation . Collectively, these data suggest that the inoculation of macrophages with C . jejuni results in the processing of IL-1 beta and apoptosis through different regulatory pathways . Furthermore, these data argue that C . jejuni may use a mechanism distinct from Salmonella typhimurium and Shigella flexneri to initiate macrophage apoptosis and release of IL-1 beta. Community Dent Oral Epidemiol, 2003 Dec, 31(6), 417 - 25 Occurrence of periodontal bacteria in healthy children: a 2-year longitudinal study; Ooshima T et al.; OBJECTIVES: To examine the occurrence of specific periodontal bacteria in children and adolescents . METHODS: Ten putative periodontal bacteria were longitudinally examined in plaque and saliva samples from 119 periodontally healthy children (2-15 years old) using a polymerase chain reaction method . RESULTS: Capnocytophaga ochracea, C . sputigena, and Actinobacillus actinomycetemcomitans were frequently found in saliva, and tended to persist in saliva for the remainder of the study, whereas Porphyromonas gingivalis, Treponema denticola, and Prevotella intermedia were rarely detected . P . nigrescens was more frequently detected in plaque and its prevalence increased with age . Eikenella corrodens and Campylobacter rectus were sometimes detected in both plaque and saliva, while Tannerella forsythensis was occasionally detected in saliva . CONCLUSION: A . actinomycetemcomitans, C . ochracea, C . sputigena, P . nigrescens, C . rectus, and E . corrodens are common members of the oral microbial flora of healthy children, whereas P . gingivalis, P . intermedia, and T . denticola appear to be transient organisms. J Biol Chem, 2004 May 7, 279(19), 20327 - 38 Epub 2004 Feb 25. Genome-wide expression analyses of Campylobacter jejuni NCTC11168 reveals coordinate regulation of motility and virulence by flhA; Carrillo CD et al.; We examined two variants of the genome-sequenced strain, Campylobacter jejuni NCTC11168, which show marked differences in their virulence properties including colonization of poultry, invasion of Caco-2 cells, and motility . Transcript profiles obtained from whole genome DNA microarrays and proteome analyses demonstrated that these differences are reflected in late flagellar structural components and in virulence factors including those involved in flagellar glycosylation and cytolethal distending toxin production . We identified putative sigma(28) and sigma(54) promoters for many of the affected genes and found that greater differences in expression were observed for sigma(28)-controlled genes . Inactivation of the gene encoding sigma(28), fliA, resulted in an unexpected increase in transcripts with sigma(54) promoters, as well as decreased transcription of sigma(28)-regulated genes . This was unlike the transcription profile observed for the attenuated C . jejuni variant, suggesting that the reduced virulence of this organism was not entirely due to impaired function of sigma(28) . However, inactivation of flhA, an important component of the flagellar export apparatus, resulted in expression patterns similar to that of the attenuated variant . These findings indicate that the flagellar regulatory system plays an important role in campylobacter pathogenesis and that flhA is a key element involved in the coordinate regulation of late flagellar genes and of virulence factors in C . jejuni. Infect Immun, 2004 Mar, 72(3), 1715 - 24 Conservation and diversity of sap homologues and their organization among Campylobacter fetus isolates; Tu ZC et al.; Campylobacter fetus surface layer proteins (SLPs), encoded by sapA homologues, are important in virulence . In wild-type C . fetus strain 23D, all eight sapA homologues are located in the 54-kb sap island, and SLP expression reflects the position of a unique sapA promoter in relation to the sapA homologues . The extensive homologies in the sap island include both direct and inverted repeats, which allow DNA rearrangements, deletion, or duplication; these elements confer substantial potential for genomic plasticity . To better understand C . fetus sap island diversity and variation mechanisms, we investigated the organization and distribution of sapA homologues among 18 C . fetus strains of different subspecies, serotypes, and origins . For all type A strains, the boundaries of the sap island were relatively consistent . A 187-bp noncoding DNA insertion near the upstream boundary of the sap island was found in two of three reptile strains studied . The sapA homologue profiles were strain specific, and six new sapA homologues were recognized . Several homologues from reptile strains are remarkably conserved in relation to their corresponding mammalian homologues . In total, the observed differences suggest that the sap island has evolved differing genotypes that are plastic, perhaps enabling colonization of varied niches, in addition to antigenic variation. J Formos Med Assoc, 2003 Dec, 102(12), 857 - 62 Comparison of rabeprazole-based four- and seven-day triple therapy and omeprazole-based seven-day triple therapy for Helicobacter pylori infection in patients with peptic ulcer; Yang KC et al.; BACKGROUND AND PURPOSE: Rabeprazole is a new proton pump inhibitor producing rapid inhibition of gastric acid secretion . This may potentiate the inhibitory effect of antibiotics against Helicobacter pylori . This study compared the efficacy, safety, and tolerability of 4- and 7-day rabeprazole-based triple therapies versus 7-day omeprazole-based triple therapy . METHODS: A total of 70 H . pylori-infected peptic ulcer patients were randomly assigned to 1 of 3 groups: RAC4 (rabeprazole 20 mg, amoxicillin 1000 mg, and clarithromycin 500 mg twice daily for 4 days), RAC7 (rabeprazole 20 mg, amoxicillin 1000 mg, and clarithromycin 500 mg twice daily for 7 days), and OAC7 (omeprazole 20mg, amoxicillin 1000 mg, and clarithromycin 500 mg twice daily for 7 days) . Endoscopy, Campylobacter-like organism (CLO) test, H . pylori culture, and 13C-urea breath test were performed before randomization and 8 weeks after the start of triple therapy . RESULTS: Intention-to-treat (ITT) eradication rates for the RAC4, RAC7, and OAC7 groups were 87% (20/23), 83%(19/23), and 88% (21/24), respectively, and per-protocol (PP) eradication rates were 91% (20/22), 95% (19/20), and 100% (21/21), respectively . There was no significant difference among the ITT or PP eradication rates of the 3 groups . All 3 regimens were well tolerated and compliance was excellent . CONCLUSIONS: One-week RAC and 1-week OAC are equally effective for H . pylori eradication in peptic ulcer patients . The duration of RAC triple therapy can be shortened to 4 days without compromising its efficacy. Water Res, 2004 Mar, 38(5), 1113 - 20 Die-off of enteric bacterial pathogens during mesophilic anaerobic digestion; Horan NJ et al.; Conventionally treated sewage sludge may contain high concentrations of potentially pathogenic microorganisms and additional treatment is required to minimise the risks to health if it is to be recycled to agricultural land . Mesophilic anaerobic digestion (MAD) is the most widely used process in the UK for stabilising sludge prior to agricultural recycling, but little is known about the fate of a number of enteric pathogens as the sludge passes through the treatment processes . The aim of this study was to determine the efficiency of MAD in removing the bacterial enteric pathogens, Salmonella senftenberg, Listeria monocytogenes and Campylobacter jejuni which were added as a spike to the digester feedstock, together with the die-off of indigenous Escherichia coli already present in the sludge . The primary sludge digestion stage of MAD was found to achieve a log removal of 1.66 for E . coli, 2.23 for L . monocytogenes and 2.23 for S . senftenberg . However, the extent of die-off was a function of the numbers of pathogens in the feed and as these increased the log removal also increased . The numbers of C . jejuni were not affected by primary sludge digestion . Additional die-off was provided by secondary sludge digestion with log removals of 1.70 for E . coli, 2.10 for S . senftenberg and 0.36 for C . jejuni. J Bacteriol, 2004 Mar, 186(5), 1518 - 30 eBURST: inferring patterns of evolutionary descent among clusters of related bacterial genotypes from multilocus sequence typing data; Feil EJ et al.; The introduction of multilocus sequence typing (MLST) for the precise characterization of isolates of bacterial pathogens has had a marked impact on both routine epidemiological surveillance and microbial population biology . In both fields, a key prerequisite for exploiting this resource is the ability to discern the relatedness and patterns of evolutionary descent among isolates with similar genotypes . Traditional clustering techniques, such as dendrograms, provide a very poor representation of recent evolutionary events, as they attempt to reconstruct relationships in the absence of a realistic model of the way in which bacterial clones emerge and diversify to form clonal complexes . An increasingly popular approach, called BURST, has been used as an alternative, but present implementations are unable to cope with very large data sets and offer crude graphical outputs . Here we present a new implementation of this algorithm, eBURST, which divides an MLST data set of any size into groups of related isolates and clonal complexes, predicts the founding (ancestral) genotype of each clonal complex, and computes the bootstrap support for the assignment . The most parsimonious patterns of descent of all isolates in each clonal complex from the predicted founder(s) are then displayed . The advantages of eBURST for exploring patterns of evolutionary descent are demonstrated with a number of examples, including the simple Spain(23F)-1 clonal complex of Streptococcus pneumoniae, "population snapshots" of the entire S . pneumoniae and Staphylococcus aureus MLST databases, and the more complicated clonal complexes observed for Campylobacter jejuni and Neisseria meningitidis. Public Health Nutr, 2004 Feb, 7(1), 85 - 9 Consumption of foods by young children with diagnosed campylobacter infection - a pilot case-control study; Cameron S et al.; OBJECTIVE: To determine whether parentally reported habitual intake of specific foods differed between children with diagnosed Campylobacter jejuni infection and children of a comparison group without diagnosed infection . DESIGN, SETTING AND SUBJECTS: Information was collected from the parents or primary caregivers of South Australian children aged 1-5 years with diagnosed C . jejuni (cases, n=172) and an age- and gender-matched group of uninfected children (controls, n=173) . Frequency of consumption of 106 food and drink items was determined for the preceding two months by food-frequency questionnaire . Four children in the control group had recorded diarrhoeal episodes during the assessment period and were excluded, so 169 responses were evaluated for this group . Information was gathered on possible confounders including socio-economic status . Response frequencies were classified into three levels of consumption (rarely, weekly or daily) and statistical comparison was made by frequency of consumption of foods versus the 'rarely' classification for cases and controls, respectively . RESULTS: Frequency of consumption of most foods, including starchy foods and fruits and vegetables, did not differ between cases and controls . However, reported consumption of eight food items (block and processed cheese (slices and spread), salami/fritz (a form of processed sausage), chicken nuggets, pasteurised milk, fish (canned or fresh) and hot French fries) was significantly higher by controls . CONCLUSIONS: The hypothesis that reported consumption of starchy foods was lower by cases than by controls was not supported by the data . However, consumption of some processed and unprocessed foods was higher by controls . Some of these foods have established bactericidal actions in vitro that may indicate a possible mechanism for this apparent protection. J Food Prot, 2004 Feb, 67(2), 239 - 45 Distribution and characterization of Campylobacter spp . from Russian poultry; Stern NJ et al.; The distribution of Campylobacter spp . on 13 poultry farms (broiler chicken, quail, pheasant, peacock, and turkey) from eight regions (Vladimir, Vologda, Voronezh, Kaluga, Liptsk, Moscow, Orenburg, and Orel) in Russia was surveyed . Intestinal materials were plated onto Campylobacter-selective medium and plates were incubated microaerobically at 42 degrees C for 24 or 48 h . Identification was based on colonial morphology, microscopic examination, and biochemical tests; latex agglutination assays were used for confirmation . In total, 116 isolates were derived from 370 samples . Isolation rates were similar, regardless of whether the birds were from small or large broiler production farms . Susceptibility of 48 representative (from these production sources) strains of Campylobacter spp . to 38 antimicrobial compounds was determined by disk diffusion assays . All strains tested were sensitive to amikacin, gentamycin, sisomycin, chloramphenicol, imipenem, oleandomycin, erythromycin, azitromycin, and ampicillin . The strains were also sensitive to 100 microg/disk of carbenicillin, fluoroquinolones, and to nitrofurans . Fluoroquinolone sensitivity was most notable and may be related to its limited application in poultry production within Russia . Hippurate and ribosomal RNA gene primers were developed and used to distinguish Campylobacter jejuni and Campylobacter coli and to provide a measure of strain discrimination . The combination of PCR analysis and randomly amplified polymorphic DNA (RAPD) typing were conducted for selected isolates . The various poultry species and the different locations yielded Campylobacter isolates with discrete randomly amplified polymorphic DNA patterns . The distribution and substantial diversity of Campylobacter spp . isolates appears similar to that previously reported in other countries. J Food Prot, 2004 Feb, 67(2), 235 - 8 Effect of intestinal content contamination on broiler carcass Campylobacter counts; Berrang ME et al.; Intestinal contents may contaminate broiler carcasses during processing . The objective of this study was to determine what effect various levels of intestinal contents had on the numbers of Campylobacter detected in broiler carcass rinse samples . Eviscerated broiler carcasses were collected from the shackle line in a commercial processing plant immediately after passing through an inside/outside washer . Broiler carcasses were cut longitudinally into contralateral halves using a sanitized saw . Cecal contents from the same flock were collected, pooled, homogenized, and used to contaminate carcass halves . Paired carcass halves were divided into groups of eight each, and then cecal contents (2, 5, 10, 50, or 100 mg) were placed onto one randomly selected half of each carcass, while the corresponding half of the same broiler carcass received no cecal contents . Campylobacter counts from carcass halves with cecal contamination were compared to the uncontaminated halves of the same carcasses using a paired t test . Carcass halves with 5 mg or more of surface cecal contamination had significantly higher numbers of Campylobacter than those without (P < 0.01) . Carcass halves contaminated with only 5 mg of cecal contents had an average of 3.3 log CFU Campylobacter per ml of rinse, while corresponding uncontaminated carcass halves had 2.6 log CFU Campylobacter per ml of rinse . These data indicate that even small (5 mg) amounts of cecal contents can cause a significant increase in the numbers of Campylobacter on eviscerated broiler carcasses . Therefore, it is important to keep such contamination to a minimum during processing. J Food Prot, 2004 Feb, 67(2), 228 - 34 Characterization of Campylobacter isolates recovered from clinically healthy pigs and from sporadic cases of campylobacteriosis in humans; Guevremont E et al.; 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 assays to detect production of cytotoxins, enterotoxins, and hemolysins . Among a subset of isolates (n = 10), up to five colonies from the same animal were characterized by PFGE . In 5 (50%) of 10 of the isolates, more than one genetic profile was observed per pig . Among the 100 isolates from pigs selected for further analysis, 81 different genetic profiles were observed, whereas 20 different genetic profiles were found among the 24 isolates of human origin . Cytotoxicity on Chinese hamster ovary cells was observed in 11 (11%) of 98 isolates from pigs and in 5 (21%) of 24 Campylobacter isolates from humans . No enterotoxin production was detected in Campylobacter isolates in this study, but 17 (71%) of 24 human and 61 (63%) of 97 pig isolates showed hemolytic activity . The study of genotypic and phenotypic profiles of swine and human isolates revealed no epidemiological relationship between isolates . The low genomic relatedness observed between groups of isolates and the weak toxicity level of swine isolates suggest that the hazard of contamination of humans by Campylobacter associated with swine production is low. J Microbiol Methods, 2004 Mar, 56(3), 395 - 400 Detection of PCR amplicons from bacterial pathogens using microsphere agglutination; Wu SJ et al.; For rapid and inexpensive detection of polymerase chain reaction (PCR) amplicons, a novel microsphere agglutination assay has been developed . PCR is carried out using biotinylated forward and reverse primers, and the amplified DNA fragments are able to agglutinate streptavidin-coated microspheres (5.7 microm in diameter) . Purification of PCR amplicons is unnecessary when initial primer concentrations are 250 nM . Agglutination can be identified visually within 2 min without any additional equipment or reagents . Using listeriolysin (lisA)-specific biotinylated primers, we have successfully detected and identified Listeria monocytogenes lisA+ cells among Salmonella typhimurium, Staphylococcus aureus, Campylobacter jejuni and Escherichia coli O157:H7 cells . The simplicity of this protocol considerably reduces the time and cost of diagnostic PCR experiments . This procedure is potentially useful for various studies and field applications. J Appl Microbiol, 2004, 96(3), 499 - 509 Microbiological examination of ready-to-eat cold sliced meats and pâté from catering and retail premises in the UK; Elson R et al.; AIMS: To establish the microbiological quality of cold ready-to-eat sliced meats and pate from catering and retail premises, and investigate links hypothesized between foodborne Campylobacter infection and the consumption of cold sliced meats . METHODS AND RESULTS: A total of 4078 cold meat and pate samples were collected and examined according to a standardized protocol . Comparison with published microbiological guidelines revealed that most ready-to-eat meat and pate samples (75%) were of satisfactory/acceptable microbiological quality and 25% were of unsatisfactory/unacceptable quality . Two cold meat samples (<1%) were of unacceptable microbiological quality because of the presence of Campylobacter jejuni in 25 g and Listeria monocytogenes at 3.4 x 104 CFU g-1 . CONCLUSIONS: Acceptable microbiological quality was associated with premises where the management was trained in food hygiene and those that had hazard analysis in place . Poor microbiological quality was associated with storage above 8 degrees C, presliced meats, infrequent cleaning of slicing equipment and poor control of practices that may lead to cross contamination . SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides important information about the microbiological quality of cold ready-to-eat meats and pate . It also assists caterers, retailers, enforcement officers and policy makers to understand how different food safety practices affect microbiological quality. J Biol Chem, 2004 Apr 16, 279(16), 16214 - 22 Epub 2004 Feb 11. The FlgS/FlgR two-component signal transduction system regulates the fla regulon in Campylobacter jejuni; Wosten MM et al.; The human pathogen Campylobacter jejuni is a highly motile organism that carries a flagellum on each pole . The flagellar motility is regarded as an important trait in C . jejuni colonization of the intestinal tract, however, the knowledge of the regulation of this important colonization factor is rudimentary . We demonstrate by phosphorylation assays that the sensor FlgS and the response regulator FlgR form a two-component system that is on the top of the Campylobacter flagellum hierarchy . Phosphorylated FlgR is needed to activate RpoN-dependent genes of which the products form the hook-basal body filament complex . By real-time reverse transcriptase-PCR we identified that FlgS, FlgR, RpoN, and FliA belong to the early flagellar genes and are regulated by sigma70 . FliD and the putative anti-sigma-factor FlgM are regulated by a sigma54- and sigma28-dependent promoters . Activation of the fla regulon is growth phase-dependent, a 100-fold rpoN mRNA reduction is seen in the early stationary phase compared with the early logarithmic phase . Whereas flaB transcription decreases, flaA transcription increases in early stationary phase . Our data show that the C . jejuni flagellar hierarchy largely differs from that of other bacteria . Phenotypical analysis revealed that unflagellated C . jejuni mutants grow three times faster in broth medium compared with wild-type bacteria . In vivo the C . jejuni flagella are needed to pass the gastrointestinal tract of chickens, but not to colonize the ceaca of the chicken. FEBS Lett, 2004 Feb 13, 559(1-3), 136 - 40 Characterization of CJ1293, a new UDP-GlcNAc C6 dehydratase from Campylobacter jejuni; Creuzenet C; Campylobacter jejuni encodes numerous sugar-nucleotide-modifying enzymes potentially involved in the biosynthesis of surface carbohydrates . One of them, CJ1293, is involved in flagellin glycosylation but its biochemical activity remains unknown . Using over-expressed and purified protein, we demonstrate that CJ1293 has UDP-GlcNAc-specific C(6) dehydratase activity . Catalysis occurs without addition of cofactor, suggesting internal recycling of NAD(P)(+) . The K(m) for UDP-GlcNAc of 50 microM indicates that CJ1293 has higher affinity for its substrate than previously characterized homologues . Based on enzymatic data, we propose that CJ1293 catalyzes the first step in the biosynthesis of bacillosamine, a sugar found in C . jejuni's protein glycosylation motifs. Epidemiol Infect, 2003 Dec, 131(3), 1181 - 6 Antibiotic resistance of Campylobacter in raw retail chickens and imported chicken portions; Wilson IG; Campylobacter isolates from raw retail chickens (n = 434) sampled between 1998 and 2000 were tested for resistance to 12 antibiotics . Among 208 campylobacters tested, more than 90% of isolates were susceptible to 4 out of 9 antibiotics (nalidixic acid, erythromycin, chloramphenicol and gentamicin) . Most campylobacters were resistant to 3 antibiotics and multiple resistance was found in 4% . Ciprofloxacin resistance was 11% . Campylobacter contamination (28%) in imported chickens (n = 150) was almost half that found in local whole chickens (50%), but the resistance of imported isolates (n = 42) was similar to that of local campylobacters . Resistance in isolates from imported chicken breasts was generally more common, but to only 4 antibiotics . Resistance patterns of chicken isolates were compared to human clinical isolates (n = 494), and a greater similarity was found between the clinical and local isolates than with imported campylobacters . Lower chloramphenicol resistance was found in clinical Campylobacter isolates than in those from chicken sources. Epidemiol Infect, 2003 Dec, 131(3), 1169 - 80 Routes for Campylobacter contamination of poultry meat: epidemiological study from hatchery to slaughterhouse; Herman L et al.; 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 hill) and on the footwear of the farmer . The administration of antibiotics did not significantly reduce Campylobacter shedding . With the exception of one flock during rearing and a few flocks in the slaughterhouse with a mixed Campylobacter coli-Campylobacter jejuni infection, C . jejuni exclusively was found both during rearing and on the carcasses . A significant correlation exits between the contamination of the broilers during rearing and the carcasses after processing . No slaughterhouse was able to avoid contamination of carcasses when status-positive animals were delivered . Moreover, six negatively delivered flocks yielded positive carcasses, the result of a supplementary contamination, which occurred during transport and slaughtering. Eur J Oral Sci, 2004 Feb, 112(1), 33 - 41 Gingival crevice microbiota from Chinese patients with gingivitis or necrotizing ulcerative gingivitis; Gmur R et al.; The objective of this study was to quantitatively compare the bacterial population structure in plaque from the gingival margin of two groups of 21 Chinese patients with gingivitis or necrotizing ulcerative gingivitis (NUG) . Subjects were recruited in four dental clinics in Eastern China . Samples were quantitatively assessed by immunofluorescence and fluorescent in situ hybridization for taxa known to be associated with periodontal diseases . The analyses showed that the fusiform taxa (Fusobacterium nucleatum/Fusobacterium periodonticum, Leptotrichia buccalis, Tannerella forsythensis, and Capnocytophaga sp.), Campylobacter rectus, Prevotella intermedia, Prevotella nigrescens, Selenomonas sputigena, and treponemes were present in both groups with high prevalence . Porphyromonas gingivalis and Actinomyces gerencseriae were much more prevalent in the NUG group . Quantitatively, most taxa, including P . gingivalis, F . nucleatum and the treponemes, accounted, on average, for < 3% of the total bacterial cell number . Only P . intermedia/P . nigrescens, P . gingivalis, S . sputigena, A . gerencseriae, and the sum of all monitored suspected periodontal pathogens were significantly increased in the NUG group . The present study demonstrates for both groups a highly diverse plaque composition and suggests that, etiologically, the overall concentration and the concerted effects of the entire group of opportunistic pathogens thriving in NUG-associated plaque are of prime importance. Am J Epidemiol, 2004 Feb 15, 159(4), 406 - 12 Household outbreaks among culture-confirmed cases of bacterial gastrointestinal disease; Ethelberg S et al.; To examine the general frequency of household outbreaks, the authors performed a retrospective search among cases of the five most frequent gastrointestinal bacterial pathogens in Denmark, a country of 5.3 million inhabitants . This was done for 57,667 cases registered from 1991 to 2001 by finding all cases that shared addresses and became infected within 3 weeks of one another . The percentage of cases that were part of household outbreaks was found to be 3.2% for Campylobacter, 13.3% for Salmonella serotype Enteritidis, 5.6% for Salmonella serotype Typhimurium, 2.0% for Yersinia enterocolitica, and 10.4% for Shigella sonnei . The vast majority of the outbreaks had not previously been registered . The wide variation in the ability to cause household outbreaks among the different types of bacteria reflects differences in their epidemiology and most likely also mirrors their overall outbreak potential . Differences in the time occurring between infections of household members may also indicate differences in the importance of person-to-person transmission for the different types of bacteria . The fact that household outbreaks occur with a relatively high frequency may be utilized in future analyses of sources of infection, in particular of Campylobacter, for which more household outbreaks than expected were identified. Joint Bone Spine, 2004 Jan, 71(1), 63 - 5 Campylobacter fetus septic arthritis revealing a malignancy; Rollot K et al.; Campylobacter fetus is a rare cause of bone and joint infection . We report a new case in a patient who also had endocarditis due to the same organism . This type of infection usually occurs in immunocompromised hosts . Investigations in our patient found a cancer of the prostate. J Clin Microbiol, 2004 Feb, 42(2), 819 - 21 Simultaneous presence of multiple Campylobacter species in dogs; Koene MG et al.; The prevalence of coinfection of Campylobacter species in dogs was determined using four isolation methods . In 26% of the positive-testing stools, multiple Campylobacter species were identified . The use of multiple isolation methods as well as the time lapse between sampling and processing are important for detection of coinfection. J Clin Microbiol, 2004 Feb, 42(2), 700 - 6 Genetic characterization and antibiotic resistance of Campylobacter jejuni isolated from meats, water, and humans in Sweden; Lindmark H et al.; The incidence of Campylobacter jejuni has increased during the last decade, and today it is the leading cause of bacterial enteritis in most developed countries . Still, there is a lack of knowledge about infection routes and to what extent identified sources are responsible for spreading the bacterium to humans . The major objective of this work was to explore the genetic similarity between C . jejuni isolated from different sources . C . jejuni isolated from patients (n = 95), five types of meat (n = 71), and raw water (n = 11) during the year 2000 were subtyped by pulsed-field gel electrophoresis (PFGE) . The pulsotypes obtained after digestion with SmaI revealed not only that C . jejuni is genetically diverse but also that specific pulsotypes occur frequently . Five clusters comprising 88 of the 162 SmaI-digested isolates were obtained . After digestion with KpnI most isolates in four of the five clusters were still indistinguishable, while the fifth cluster was strongly dissolved . The clusters comprised high frequencies of human and meat isolates, while only one of nine water isolates belonged to a cluster . The largest cluster comprised 21 human isolates, one raw water isolate, and seven chicken meat isolates, originating from at least six different broiler flocks . Low frequencies of antibiotic resistance were revealed when the meat and water isolates were tested for sensitivity to six antibiotics . Interestingly, the five isolates resistant to quinolones displayed similar or identical pulsotypes . The results showed that PFGE has proved useful in identifying clones and will be used in future work focusing on identification and eradication of the major reservoirs for common clones. J Clin Microbiol, 2004 Feb, 42(2), 627 - 30 Enzyme-linked immunosorbent assay for Helicobacter pylori needs adjustment for the population investigated; Hoang TT et al.; Helicobacter pylori infection and peptic ulcer disease are common in developing countries, e.g., Vietnam . An enzyme-linked immunosorbent assay (ELISA) for screening of patients and for seroepidemiology is a useful tool but needs to be validated in the population studied . We used in-house ELISA with sonicated Swedish and Vietnamese strains as antigens to measure immunoglobulin G antibodies after absorption with sonicated Campylobacter jejuni in sera from 270 H . pylori culture-confirmed peptic ulcer patients, 128 Swedish urea-breath test and immunoblot-positive healthy controls, and 432 Vietnamese immunoblot-positive population controls . Sonicated whole-cell antigen based on the local strains showed a significantly better performance . Immunoblot-positive peptic ulcer patients had significantly higher antibody concentrations than immunoblot-positive population controls, necessitating a lower cutoff level if serology is used for screening or epidemiological purposes . The study shows that the parameters of ELISA for H . pylori need to be adjusted for the population being investigated. Appl Environ Microbiol, 2004 Feb, 70(2), 1182 - 9 Comparison of survival of Campylobacter jejuni in the phyllosphere with that in the rhizosphere of spinach and radish plants; Brandl MT et al.; Campylobacter jejuni has been isolated previously from market produce and has caused gastroenteritis outbreaks linked to produce . We have tested the ability of this human pathogen to utilize organic compounds that are present in leaf and root exudates and to survive in the plant environment under various conditions . Carbon utilization profiles revealed that C . jejuni can utilize many organic acids and amino acids available on leaves and roots . Despite the presence of suitable substrates in the phyllosphere and the rhizosphere, C . jejuni was unable to grow on lettuce and spinach leaves and on spinach and radish roots of plants incubated at 33 degrees C, a temperature that is conducive to its growth in vitro . However, C . jejuni was cultured from radish roots and from the spinach rhizosphere for at least 23 and 28 days, respectively, at 10 degrees C . This enteric pathogen also persisted in the rhizosphere of spinach for prolonged periods of time at 16 degrees C, a temperature at which many cool-season crops are grown . The decline rate constants of C . jejuni populations in the spinach and radish rhizosphere were 10- and 6-fold lower, respectively, than on healthy spinach leaves at 10 degrees C . The enhanced survival of C . jejuni in soil and in the rhizosphere may be a significant factor in its contamination cycle in the environment and may be associated with the sporadic C . jejuni incidence and campylobacteriosis outbreaks linked to produce. Appl Environ Microbiol, 2004 Feb, 70(2), 822 - 30 Genotypic and antibiotic susceptibility characteristics of a Campylobacter coli population isolated from dairy farmland in the United Kingdom; Leatherbarrow AJ et al.; Campylobacter infections are the most common cause of bacterial enteritis in humans, and nearly 8% of such infections are caused by Campylobacter coli . Most studies have concentrated on Campylobacter jejuni, frequently isolated from intensively farmed poultry and livestock production units, and few studies have examined the spread and relatedness of Campylobacter across a range of geographical and host boundaries . Systematic sampling of a 100-km2 area of mixed farmland in northwest England yielded 88 isolates of C . coli from a range of sample types and locations, and water was heavily represented . Screening for antibiotic resistance revealed a very low prevalence of resistance, while genotyping performed by using three methods (flaA PCR restriction fragment length polymorphism {RFLP}, pulsed-field gel electrophoresis {PFGE}, and fluorescent amplified fragment length polymorphism {fAFLP}) provided insights into the genomic relatedness of isolates from different locations and hosts . Isolates were classified into 23 flaA groups, 34 PFGE groups, and five major fAFLP clusters . PFGE banding analysis revealed a high level of variability and no clustering by sample type . fAFLP and flaA analyses successfully grouped the isolates by sample type . We report preliminary findings suggesting that there is a strain of C . coli which may have become adapted to survival or persistence in water and that there is a group of mainly water-derived isolates from which unusual flaA PCR fragments were recovered. J Clin Periodontol, 2003 Nov, 30(11), 1003 - 10 Relationship between periodontal pocket sulfide levels and subgingival species; Torresyap G et al.; BACKGROUND: Many species implicated in the pathogenesis of periodontal disease produce volatile sulfur compounds (VSC) . This investigation examined the relationship between levels of sulfide and subgingival bacterial species in the same periodontal pockets . MATERIAL AND METHODS: Twenty chronic periodontitis subjects were measured clinically at six sites per tooth for plaque, gingivitis, bleeding on probing, suppuration, pocket depth and attachment level . Subgingival plaque samples, taken from the mesial aspect of each tooth, were individually analyzed for their content of 40 bacterial species using checkerboard DNA-DNA hybridization . Sulfide levels were measured at the same sites using a Diamond Probe/Perio 2000 system . Clinical and microbiological data were averaged for sulfide-positive and -negative sites separately in each subject and then averaged across subjects . Significance differences in clinical and microbial parameters between sulfide-positive and -negative sites were sought using the Wilcoxon signed ranks test . RESULTS: Mean total DNA probe counts (x10(5), +/-SEM) at sulfide-negative and -positive sites were 44.0 +/- 9.9 and 65.0 +/- 13.3, respectively (p < 0.01) . Seventeen species were found at significantly higher levels in sulfide-positive than -negative sites . These included abundant producers of VSC such as members of the genera Fusobacterium, Campylobacter, Prevotella, Treponema and Eubacterium, and Bacteriodes forsythus, Selenomonas noxia and Propionibacterium acnes . Prevotella intermedia, Bacteriodes forsythus, Prevotella nigrescens, Fusobacterium nucleatum ss vincentii and Treponema denticola exhibited the greatest difference in mean counts between sulfide-negative and -positive sites . Orange and red complex species were at higher counts at shallow (< 4 mm) sulfide-positive than shallow sulfide-negative sites . Although not statistically significant, mean clinical parameters were somewhat higher at sulfide-positive than sulfide-negative sites . CONCLUSIONS: Intra-pocket sulfide levels reflect the levels of sulfide-producing species and may provide useful diagnostic information. J Clin Periodontol, 2003 Nov, 30(11), 990 - 5 Bacterial colonization during de novo plaque formation; Ramberg P et al.; OBJECTIVE: To determine microbial changes that occur during plaque formation in a dentition free of gingival inflammation . MATERIAL AND METHODS: Ten subjects were recruited . The study included one preparatory period (2 weeks) and a plaque accumulation period (4 days) . The volunteers exercised proper tooth cleaning methods, were scaled and received repeated professional mechanical tooth cleaning during the preparatory period . During the plaque accumulation period, the participants abstained from plaque control measures . Plaque was scored on the approximal surfaces of maxillary and mandibular premolars on Days 0, 1, 2 and 4 using a scale from 0 to 5 and according to the criteria of the Quigley and Hein Plaque Index (QHI) . Supragingival plaque samples were obtained from the same intervals and surfaces and evaluated using a checkerboard DNA-DNA hybridization technique . RESULTS: The mean QHI increased from 0 to 1.6 (Day 4) . The total number of organisms on Day 0 averaged 140 x 10(5) and increased to about 210 x 10(5) after 4 days without oral hygiene . The most dominant species on Day 0 were members of the genus Actinomyces . These organisms comprised almost 50% of the microbiota evaluated . None of the Actinomyces species increased significantly during the 4 days . Some Streptococcus species increased significantly over time as well as species of the genera Capnocytophaga, Campylobacter, Fusobacteria and Actinomyces actinomycetemcomitans . CONCLUSION: In the present investigation, the preparatory phase established a situation with minimal gingival inflammation and close to zero amounts of dental plaque . The Day 0 plaque samples exhibited high proportions of Actinomyces species . During the 4 days of no oral hygiene, there was a small increase in total numbers of organisms as well as a modest increase in the proportion of "disease-associated" taxa such as species of the "orange complex" species. Int J Food Microbiol, 2004 Feb 1, 90(3), 321 - 9 Emergence of variants with altered survival properties in stationary phase cultures of Campylobacter jejuni; Martinez-Rodriguez A et al.; During the stationary phase of Campylobacter jejuni NCTC 11351 viable numbers fluctuate in a characteristic fashion . After reaching the maximum cell count (ca . 2 x 10(9) CFU/ml) in early stationary phase (denoted phase 1), viable numbers subsequently decrease to about 10(6) CFU/ml after 48 h and then increase again to about 10(8) CFU/ml (denoted phase 2) before decreasing once more to a value intermediate between the previous maximum and minimum values . To investigate whether the increase in viable numbers following the initial decline was due to the emergence of a new strain with a growth advantage in stationary phase analogous to the 'GASP' phenotype described in Escherichia coli {Science 259 (1993) 1757}, we conducted mixed culture experiments with cells from the original culture and antibiotic-resistant marked organisms isolated from the re-growth phase . In many experiments of this type, strains isolated from phase 2 failed to out-compete the original strain and we have thus been unable to demonstrate a convincing GASP phenotype . However, strains isolated from phase 2 showed a much lower rate of viability loss in early stationary phase and a small increase in resistance to aeration, peroxide challenge and heat, indicating that the emergent strain was different from the parent . These results support the view that dynamic population changes occur during the stationary phase of C . jejuni that may play a role in the survival of this organism. Anal Chem, 2004 Feb 1, 76(3), 619 - 26 Selective detection and identification of sugar nucleotides by CE-electrospray-MS and its application to bacterial metabolomics; Soo EC et al.; A novel method employing CE-ESMS and precursor ion scanning was developed for the selective detection of nucleotide-activated sugars . By using precursor ion scanning for fragment ions specific to the different nucleotide carriers, i.e., ions at m/z 322 for cytidine monophosphate, m/z 323, 385, and 403 for uridine diphosphate, m/z 362, 424, and 442, for guanosine diphosphate, and m/z 346, 408, and 426 for adenosine diphosphate, it was possible to selectively detect sugar nucleotides involved in the biosynthesis of glycoconjugates such as glycoproteins and lipopolysaccharides . Enhancement of sensitivity was achieved using N-(2-hydroxyethyl)piperazine-N'-(2-ethanesulfonic acid) as a sample stacking buffer and provided detection limits between 0.2 and 3.8 pmol.mL(-)(1) . The present CE-ESMS method provided linear dynamic ranges over the concentrations 0.2-164 nM (r(2) = 0.952-0.997) for different nucleotide sugar standards . The application of this method is demonstrated for the identification of intracellular pools of sugar nucleotides in wild type and isogenic mutants from the bacterial pathogen Campylobacter jejuni . By using product ion scanning (with and without front-end collision-induced dissociation), it was possible to determine the precise nature of unexpected sugar nucleotides involved in the biosynthesis of pseudaminic acid, a sialic acid-like sugar previously observed on the flagellin of some pathogenic bacteria. Infect Immun, 2004 Feb, 72(2), 1199 - 203 Comparison of Campylobacter jejuni isolates implicated in Guillain-Barré syndrome and strains that cause enteritis by a DNA microarray; Leonard EE 2nd et al.; We asked whether Campylobacter jejuni isolated from patients with Guillain-Barre syndrome (GBS) differ from isolates isolated from patients with uncomplicated gastrointestinal infection using DNA microarray analysis . We found that specific GBS genes or regions were not identified, and microarray analysis confirmed significant genomic heterogeneity among the isolates. Infect Immun, 2004 Feb, 72(2), 1162 - 5 Evidence for acquisition of the lipooligosaccharide biosynthesis locus in Campylobacter jejuni GB11, a strain isolated from a patient with Guillain-Barré syndrome, by horizontal exchange; Gilbert M et al.; Campylobacter jejuni GB11, a strain isolated from a patient with Guillain-Barre syndrome, has been shown to be genetically closely related to the completely sequenced strain C . jejuni NCTC 11168 by various molecular typing and serotyping methods . However, we observed that the lipooligosaccharide (LOS) biosynthesis genes strongly diverged between GB11 and NCTC 11168 . We sequenced the LOS biosynthesis locus of GB11 and found that it was nearly identical to the class A LOS locus from the C . jejuni HS:19 Penner serotype strain (ATCC 43446) . Analysis of the DNA sequencing data showed that a horizontal exchange event involving at least 14.26 kb had occurred in the LOS biosynthesis locus of GB11 between galE (Cj1131c in NCTC 11168) and gmhA (Cj1149 in NCTC 11168) . Mass spectrometry of the GB11 LOS showed that GB11 expressed an LOS outer core that mimicked the carbohydrate portion of the gangliosides GM1a and GD1a, similar to C . jejuni ATCC 43446 . The serum from the GB11-infected patient was shown to react with the LOS from both GB11 and ATCC 43446 but not with that from NCTC 11168 . These data indicate that the antiganglioside response in the GB11-infected patient was raised against the structures synthesized by the acquired class A LOS locus. Infect Immun, 2004 Feb, 72(2), 1116 - 25 Gastroenteritis in NF-kappaB-deficient mice is produced with wild-type Camplyobacter jejuni but not with C . jejuni lacking cytolethal distending toxin despite persistent colonization with both strains; Fox JG et al.; Campylobacter jejuni continues to be a leading cause of bacterial enteritis in humans . However, because there are no readily available animal models to study the pathogenesis of C . jejuni-related diseases, the significance of potential virulence factors, such as cytolethal distending toxin (CDT), in vivo are poorly understood . Mice deficient in NF-kappaB subunits (p50(-/-) p65(+/-)) in a C57BL/129 background are particularly susceptible to colitis induced by another enterohepatic microaerobe, Helicobacter hepaticus, which, like C . jejuni, produces CDT . Wild-type C . jejuni 81-176 and an isogenic mutant lacking CDT activity (cdtB mutant) were inoculated into NF-kappaB-deficient (3X) and C57BL/129 mice . Wild-type C . jejuni colonized 29 and 50% of the C57BL/129 mice at 2 and 4 months postinfection (p.i.), respectively, whereas the C . jejuni cdtB mutant colonized 50% of the C57BL/129 mice at 2 p.i . but none of the mice at 4 months p.i . Although the C57BL/129 mice developed mild gastritis and typhlocolitis, they had robust immunoglobulin G (IgG) and Th1-promoted IgG2a humoral responses to both the wild-type strain and the C . jejuni cdtB mutant . In contrast, 75 to 100% of the 3X mice were colonized with both the wild type and the C . jejuni cdtB mutant at similar levels at all times examined . Wild-type C . jejuni caused moderately severe gastritis and proximal duodenitis in 3X mice that were more severe than the gastrointestinal lesions caused by the C . jejuni cdtB mutant . Persistent colonization of NF-kappaB-deficient mice with the wild type and the C . jejuni cdtB mutant was associated with significantly impaired IgG and IgG2a humoral responses (P < 0.001), which is consistent with an innate or adaptive immune system defect(s) . These results suggest that the mechanism of clearance of C . jejuni is NF-kappaB dependent and that CDT may have proinflammatory activity in vivo, as well as a potential role in the ability of C . jejuni to escape immune surveillance . NF-kappaB-deficient mice should be a useful model to further study the role of CDT and other aspects of C . jejuni pathogenesis. Infect Immun, 2004 Feb, 72(2), 916 - 22 In vivo phase variation and serologic response to lipooligosaccharide of Campylobacter jejuni in experimental human infection; Prendergast MM et al.; Some Campylobacter jejuni strains which exhibit mimicry of gangliosides in their lipooligosaccharides (LOSs) are associated with development of Guillain-Barre syndrome, which complicates the selection of a suitable C . jejuni strain in a live-attenuated vaccine . C . jejuni 81-176 is the most well characterized strain available, but structurally, LOS of C . jejuni 81-176 exhibits mimicry of predominantly GM2 and GM3 gangliosides . We compared the antiganglioside human serologic responses of 22 volunteers post-oral vaccination (two-dose series, 14 days apart) with a killed whole-cell C . jejuni vaccine, those of volunteers (22 following initial challenge and 5 upon rechallenge) experimentally infected with the homologous C . jejuni vaccine strain 81-176, and those of 12 volunteers used as controls (placebo recipients) . All volunteers were evaluated using thin-layer chromatography immuno-overlay and a panel of nine gangliosides at days 0, 21, and 28 either postvaccination or postinoculation . Antiganglioside antibodies were identified at baseline in 6 of the 61 volunteers (9.8%) . There were no antiganglioside antibodies observed following vaccination or experimental infection rechallenge . Evidence of seroconversion was observed in 2 of 22 (9.1%) in the initial infection challenge group, comparable to 1 of 12 (8.3%) in the placebo recipients . Additional testing of seven selected volunteers in the initial challenge group at days 0, 3, 7, 10, 21, 28, and 60 showed that when antiganglioside antibodies occurred (mostly anti-GM1 and -GM2), responses were weak and transient . Furthermore, evidence from serologic probing of LOSs of isolates recovered from stools of six volunteers indicated that the isolates had undergone antigenic phase variation in ganglioside mimicry during passage in vivo . Collectively, with the exception of one volunteer with anti-GM2 antibodies at day 60, the results show an absence of persistent antiganglioside antibodies after experimental infection with C . jejuni or following administration of a killed C . jejuni whole-cell oral vaccine, although LOS phase variation occurred. J Dairy Sci, 2003 Dec, 86(12), 3849 - 56 Microbiological quality of raw goat's and ewe's bulk-tank milk in Switzerland; Muehlherr JE et al.; A total of 407 samples of bulk-tank milk (344 of goat's milk and 63 of ewe's milk) collected from 403 different farms throughout Switzerland, was examined . The number of farms investigated in this study represents 8% of the country's dairy-goat and 15% of its dairy-sheep farms . Standard plate counts and Enterobacteriaceae counts were performed on each sample . Furthermore, the prevalence of Staphylococcus aureus, Campylobacter spp., Shiga toxin-producing Escherichia coli, Salmonella spp., and Mycobacterium avium ssp . paratuberculosis was studied . The median standard plate count for bulk-tank milk from small ruminants was 4.70 log cfu/ml (4.69 log cfu/ml for goat's milk and 4.78 log cfu/ml for ewe's milk), with a minimum of 2.00 log cfu/ml and a maximum of 8.64 log cfu/ml . Enterobacteriaceae were detected in 212 (61.6%) goat's milk and 45 (71.4%) ewe's milk samples, whereas S . aureus was detected in 109 (31.7%) samples of goat's milk and 21 (33.3%) samples of ewe's milk . Campylobacter spp . and Salmonella spp . were not isolated from any of the samples . However, 16.3% of the goat's milk and 12.7% of the ewe's milk samples were polymerase chain reaction (PCR)-positive for Shiga toxin-producing E . coli . Seventy-nine (23.0%) goat's tank-milk and 15 (23.8%) ewe's tank-milk samples were PCR-positive for insertion sequence 900, providing presumptive evidence for the presence of M . avium ssp . paratuberculosis . These results form the basis for determining the microbiological quality standards for goat's and ewe's milk . Moreover, the data presented form part of the risk assessment program for raw milk from small ruminants in Switzerland. J Pak Med Assoc, 2003 Nov, 53(11), 517 - 20 Frequency of Campylobacter jejuni in diarrhoea/dysentery in children in Rawalpindi and Islamabad; Ali AM et al.; OBJECTIVE: To determine the frequency of Campylobacter jejuni infection in children suffering from diarrhoea/dysentery in the Department of Microbiology, Army Medical College and Military Hospital, Rawalpindi, from 29 August 2002 to 29 November 2002 . METHODS: The study was carried out on one hundred stool samples of children up to the age of twelve years admitted with diarrhoea/dysentery in Military hospital, Rawalpindi . The samples were collected in clean polypropylene containers containing Cary Blair medium . These were transported to the Microbiology Department, Army Medical College, Rawalpindi within 1-2 hours . The samples were inoculated on Modified Preston (Oxoid) and Karmali media (Oxoid) beside other routine stool culture media . The cultures were incubated at 42 degrees C under microaerophilic conditions . The growth after 48 hours was provisionally identified by colonial morphology, oxidase test, Gram staining and motility . The organisms were identified to species level by hippurate hydrolysis, urease test, nitrate reduction, catalase test, H2S production and resistance to cephalothin . RESULTS: Eighteen percent of samples yielded the growth of Campylobacter jejuni . Mean age of children with Campylobacter jejuni infection was 18 months with peak incidence from 12 to 21 months . Male female ratio was 1.7:1 . All the children had loose motions . Seven out 18 (39%) had a combination of symptoms of loose motions, vomiting and pain abdomen . Those having fever with or without other complaints constituted 11 out of 18 (61.11%) i.e . more than 50% of all the children yielding C . jejuni had fever . About 90% of diarrhoeal stools had blood and fifty percent also had mucous . There was either history of chicken meat consumption or contact with cattle and pets in most of the cases and both in some of them . CONCLUSION: Campylobacter jejuni is a frequent cause of diarrhoea/dysentery in children in our set up . In children it is often related to pets keeping and chicken meat consumption . In the remaining, untreated drinking water may be the source . Campylobacter jejuni frequently presents with blood and mucous in stools with sporadic cases presenting with watery diarrhoea. Can J Microbiol, 2003 Nov, 49(11), 727 - 31 FeoB is not required for ferrous iron uptake in Campylobacter jejuni; Raphael BH et al.; Among strains of Campylobacter jejuni, levels of ferrous iron (Fe2+) uptake was comparable . However, C . jejuni showed a lower level of ferrous iron uptake than Escherichia coli . Consistent with studies of E . coli, Fe2+ uptake in C . jejuni was significantly enhanced by low Mg2+ concentration . The C . jejuni genome sequence contains a single known ferrous iron uptake gene, feoB, whose product shares 50% amino acid identity to Helicobacter pylori FeoB and 29% identity to E . coli FeoB . However, Fe2+ uptake could not be attributed to FeoB for several reasons . Site-directed mutations in feoB caused no defect in 55Fe2+ uptake . Among C . jejuni strains, various nucleotide alterations were found in feoB, indicating that some C . jejuni feoB genes are defective . In addition, uptake could not be attributed to the magnesium transporter CorA, since no reduction in 55Fe2+ uptake was observed in the presence of a CorA-specific inhibitor. Zhonghua Er Ke Za Zhi, 2003 Sep, 41(9), 684 - 7 {Experimental study on the therapeutic mechanism of high dose intravenous immunoglobulin in treatment of immune-mediated peripheral neuropathy}; Gao YX et al.; OBJECTIVE: To explore the therapeutic basis of high dose intravenous immunoglobulin (IVIg) in treatment of peripheral neuropathy induced by Campylobacter jejuni lipopolysaccharide (CJ LPS) . METHOD: (1) IVIg (400 mg/kg x d) was given to the rats on the different days respectively during the immunization with CJ LPS . Histological study of sciatic nerve was performed on the 35 th day after immunization . The titer of anti-CJ LPS antibody in sera of immunized rats was measured by ELISA; IgG deposition was detected by immunohistochemistry and expression of TNF-alpha mRNA in the pathological nerves by in situ hybridization histochemistry . (2) When PBMCs were stimulated by CJ LPS in vitro, IVIg was added into culture medium at the doses of 1, 2.5, 5, and 10 mg/ml, respectively . Pathological examination of sciatic nerve was performed on the 7th day after perineural injection of the supernatants . Expression of TNF-alpha mRNA in PBMCs stimulated by CJ LPS in medium was detected by in situ hybridization histochemistry after adding IVIg . RESULTS: (1) The rate of abnormal fibers appearance in IVIg group (1.0%) was much lower than that of the control group (15.0%) after immunization with CJ LPS, P < 0.01 . The titer of antibody in control group was 9 times higher than that of IVIg group . There was no expression of immunoglobulin and TNF-alphamRNA in peripheral nerves in IVIg group, but high expression was found in control group in which no IVIg was injected . (2) The expression rates of TNF-alphamRNA on the PBMCs in IVIg group (1.0%) was much lower than that of control group (9.5%) . (3) When the PBMCs of normal rats were stimulated by CJ LPS, the expression rates of TNF-alphamRNA in PBMCs of 5 mg/ml IVIg group (3.0%) or 10 mg/ml IVIg group (2.0%) were much lower than that of 1 mg/ml IVIg group (15.0%) or 2.5 mg/ml IVIg group (11.5%), P < 0.01 . The rate of abnormal fibers appearance in 5 mg/ml IVIg group (9.8%) or 10 mg/ml IVIg group (8.5%) was much lower than that of 1 mg/ml IVIg group (50.0%), 2.5 mg/ml IVIg group (41.0%) or control group (50.8%) after the perineural injection with the supernatants, respectively, P < 0.01 . CONCLUSION: The therapeutic effect of high dose IVIg might be associated with inhibition of the humoral and cellular immunity simultaneously in peripheral neuropathy induced by CJ LPS. Nat Struct Mol Biol, 2004 Feb, 11(2), 163 - 70 Epub 2004 Jan 18. Structural analysis of the sialyltransferase CstII from Campylobacter jejuni in complex with a substrate analog; Chiu CP et al.; Sialic acid terminates oligosaccharide chains on mammalian and microbial cell surfaces, playing critical roles in recognition and adherence . The enzymes that transfer the sialic acid moiety from cytidine-5'-monophospho-N-acetyl-neuraminic acid (CMP-NeuAc) to the terminal positions of these key glycoconjugates are known as sialyltransferases . Despite their important biological roles, little is understood about the mechanism or molecular structure of these membrane-associated enzymes . We report the first structure of a sialyltransferase, that of CstII from Campylobacter jejuni, a highly prevalent foodborne pathogen . Our structural, mutagenesis and kinetic data provide support for a novel mode of substrate binding and glycosyl transfer mechanism, including essential roles of a histidine (general base) and two tyrosine residues (coordination of the phosphate leaving group) . This work provides a framework for understanding the activity of several sialyltransferases, from bacterial to human, and for the structure-based design of specific inhibitors. Shi Yan Sheng Wu Xue Bao, 2003 Dec, 36(6), 453 - 8 {Effects of serum from a patient with acute motor axonal neuropathy on spinal motor neurons cultured in vitro}; Wu SY et al.; The effect of serum from a patient with acute motor axonal neuropathy (AMAN) on cultured motor neurons was studied . The ventral spinal ventral tissue was isolated from embryonic rats and digested into dissociated cell suspension for culture in vitro . The cultured cells were stained with SMI-32, a non-phosphorylated neurofilment marker monoclonal antibody to identify motor neurons . The 6 days' cultured cells were exposed to the AMAN patient serum in a concentration of 25%, and to the normal human serum as the control . Positive PennerO:19 Campylobacter jejuni lipopolysaccharide antibody in the AMAN serum used in this experiment had been testified . The serum-cultured motor neurons were observed morphologically and also stained by Guillery Shirra and Webster method . With this staining, degenerated nerve fibers were brown-black and normal nerve fibers were brown-yellow . At the 9th h after the AMAN serum exposure, the axon degenerated and was stained brown-black due to increased silver-phile property . At the 12th h, the neuron soma began to swell and nuclear deviation with silver granules depositing in the cytoplasm . At last, the neurons began to die from the 16th h of the exposure . However, the control motor neurons did not show these alterations in the same period of culture . The serum of AMAN patient may be toxic to the neurite of motor neuron and thus cause axon degeneration, then soma alterations and death followed . It is suggested that Campylobacter jejuni lipopolysaccharide antibody may play an important role in this process without the participation of macrophages and complements. Gastroenterology, 2003 Dec, 125(6), 1651 - 9 Relative importance of enterochromaffin cell hyperplasia, anxiety, and depression in postinfectious IBS; Dunlop SP et al.; BACKGROUND & AIMS: Both psychological and mucosal changes (increased enterochromaffin {EC} cells and T lymphocytes) have been associated with postinfectious irritable bowel syndrome (PI-IBS) . However, previous studies have been underpowered to determine the relative importance of these changes in predicting the development of PI-IBS . Our aim was to prospectively determine the relative importance of both psychological and histologic factors in the development of PI-IBS after Campylobacter infection . METHODS: Questionnaires detailing psychological and bowel symptoms were sent to 1977 patients 3 months after infection . Twenty-eight patients with new-onset PI-IBS, 28 age- and sex-matched patient controls who were asymptomatic after infection, and 34 healthy volunteers underwent rectal biopsy, which was assessed for serotonin-containing EC cells, mast cells, and lamina propria T lymphocytes . RESULTS: PI-IBS, predominantly of the diarrhea-predominant subtype, occurred in 103 of 747 (13.8%) of those infected . EC cell counts per high-power field (hpf) were higher in patients with PI-IBS (35.8 +/- 1.2) compared with patient controls (30.6 +/- 1.9; P = 0.022) and volunteers (29.1 +/- 1.8; P = 0.006) . Lamina propria T lymphocytes per hpf were higher in patients with PI-IBS (127.1 +/- 8.7) and patient controls (113.4 +/- 6.2) in contrast to healthy volunteers (97.1 +/- 5.7) (P = 0.006 and P = 0.058, respectively) . Anxiety, depression, and fatigue were significantly increased in patients with PI-IBS compared with patient controls . Multivariate analysis indicated that increased EC cell counts and depression were equally important predictors of developing PI-IBS (relative risk, 3.8 and 3.2 for each standard deviation increase in respective values) . CONCLUSIONS: Both increased EC cells and depression are important independent predictors of developing PI-IBS. N Engl J Med, 2004 Jan 15, 350(3), 239 - 48 Immunoproliferative small intestinal disease associated with Campylobacter jejuni; Lecuit M et al.; BACKGROUND: Immunoproliferative small intestinal disease (also known as alpha chain disease) is a form of lymphoma that arises in small intestinal mucosa-associated lymphoid tissue (MALT) and is associated with the expression of a monotypic truncated immunoglobulin alpha heavy chain without an associated light chain . Early-stage disease responds to antibiotics, suggesting a bacterial origin . We attempted to identify a causative agent . METHODS: We performed polymerase chain reaction (PCR), DNA sequencing, fluorescence in situ hybridization, and immunohistochemical studies on intestinal-biopsy specimens from a series of patients with immunoproliferative small intestinal disease . RESULTS: Analysis of frozen intestinal tissue obtained from an index patient with immunoproliferative small intestinal disease who had a dramatic response to antibiotics revealed the presence of Campylobacter jejuni . A follow-up retrospective analysis of archival intestinal-biopsy specimens disclosed campylobacter species in four of six additional patients with immunoproliferative small intestinal disease . CONCLUSIONS: These results indicate that campylobacter and immunoproliferative small intestinal disease are associated and that C . jejuni should be added to the growing list of human pathogens responsible for immunoproliferative states . Zhonghua Er Ke Za Zhi, 2003 Dec, 41(12), 934 - 9 {Damage to peripheral nerves induced by Campylobacter jejuni exotoxin}; Xie LS et al.; OBJECTIVE: To explore the pathogenesis of the damage to peripheral nerves induced by Campylobacter jejuni exotoxin (CJT) . METHODS: (1) Animal models: (1) The CJT was extracted from PEN 19-CJ and injected perineurally and intravenously to Wistar rats . (2) The sera and the supernatants of peripheral blood mononuclear cells (PBMCs), taken from the rats immunized with the CJT, were injected perineurally at sciatic nerves of experimental rats and intravenously, respectively . (2) Histopathologic study of sciatic nerves: the animals were sacrificed and their sciatic nerves were examined for tease fibers, transverse section with toluidine blues staining and electron microscopy . (3) Immunohistochemistry: sections of sciatic nerves of either normal rats or human which were incubated with CJT and the sciatic nerves with pathological changes induced by CJT were obtained for observation of the binding capability of CJT with peripheral nerves by SABC and FITC-immunofluorescence methods, and nucleic acid hybridization techniques for detection of TNF-alpha mRNA expression in pathological sciatic nerves samples . RESULTS: (1) Remarkable peripheral neuropathies with axon degeneration and/or demyelination were found in the nerves induced by both CJT injection perineurally and intravenously . The axon degeneration was more obvious . Pathological changes were identified in 76.8% (2,763/3,600) of teasing fibers after perineural injection, but only 9.6% (230/2,400) of fibers were damaged in control group (P < 0.01) . The peak severity of fiber damage was found on the 3rd day after CJT intravenous injection with the incidence of abnormal fibers was 19.5% (390/2,000), and abnormalities of 15.5% (310/2000) on the 14th day . However, no abnormal changes were demonstrated in control group (P < 0.01) . So was in the groups injected with anti-CJT sera and the supernatants of PBMCs compared with control (P > 0.05) . (2) Binding of CJT to the nerve was found dominant in the sciatic nerves taken from normal rats or human either incubated with CJT or in the pathological sciatic nerves induced by CJT to various degrees . The binding of CJT to all these nerves was determined . (3) After intravenous injection with CJT, no histopathologic change could be found in the other viscera of the rats, with the exception of remarkable pathological change in peripheral nerves . CONCLUSIONS: (1) CJT could remarkably damage the peripheral nerves in rats . Specific pathogenicity of CJT to peripheral nerves was well shown, because no histopathologic abnormalities could be found in the other viscera, such as brain, liver and kidney etc . although there was remarkable pathological change along the peripheral nerve in the animals . (2) No immunological pathogenicity of CJT could be demonstrated in the nerves of rats after immunization with CJT. Anal Chem, 2004 Jan 15, 76(2), 433 - 40 Detection of campylobacter and Shigella species in food samples using an array biosensor; Sapsford KE et al.; Campylobacter and Shigella bacteria are common causes of food- and water-borne illness worldwide . There is a current need in food, medical, environmental, and military markets for a rapid and user-friendly method of detecting such pathogens . The array biosensor developed at the NRL encompasses these qualities . In this study, 25-min, sandwich immunoassays were developed for the detection of Campylobacter and Shigella species in both buffer and a variety of food and beverage samples . The limit of detection for Shigella dysenteriae in buffer and chicken carcass wash was 4.9 x 10(4) cfu mL(-)(1), whereas Campylobacter jejuni could be measured at concentrations as low as 9.7 x 10(2) cfu mL(-)(1) . The limits of detection and dynamic range were found to vary depending on the sample matrix, but could be improved by running the sample over the waveguide surface for longer periods of time . Samples were run with no preconcentration or enrichment steps and little-to-no sample pretreatment prior to analysis. Emerg Infect Dis, 2003 Nov, 9(11), 1482 - 3 Fluoroquinolone resistance in Campylobacter absent from isolates, Australia; Unicomb L et al.; Fluoroquinolone resistance was detected in 12 of 370 Australian human Campylobacter isolates; 10 of these were travel-associated, and for 2 isolates travel status was unknown . No resistance was found in isolates known to be locally acquired . In Australia, fluoroquinolones have not been licensed for use in food production animals, a policy that may have relevance for countries with fluoroquinolone-resistant Campylobacter. Emerg Infect Dis, 2003 Nov, 9(11), 1479 - 81 Fluoroquinolone susceptibility of Campylobacter strains, Senegal; Cardinale E et al.; To assess fluoroquinolone susceptibility of Campylobacter strains in Senegal, skin samples were collected from 250 chicken carcasses from January 2001 to October 2002 . Among 205 isolated Campylobacter strains, 59% and 41% were identified as Campylobacter jejuni and C . coli, respectively; the overall ciprofloxacin-resistance rate was 34%. Poult Sci, 2003 Dec, 82(12), 1995 - 9 Presence of Campylobacter inthe respiratory tract of broiler carcasses before and after commercial scalding; Berrang ME et al.; Campylobacter could be detected in the thoraco-abdominal cavity of broiler carcasses even if they were carefully eviscerated by hand with no evidence of intestinal rupture or leakage . If Campylobacter is present in the air sacs, which are unavoidably torn during evisceration, it could contaminate the thoraco-abdominal cavity of the eviscerated carcass . This study was done to determine if Campylobacter contamination is present in the respiratory tract of broilers prior to evisceration . Whole carcass rinses and respiratory tract washes were done on broiler carcasses collected at a commercial processing plant just before and just after scalding . Samples were cultured for presence and numbers of Campylobacter, Escherichia coli, coliforms, and total aerobic bacteria . Campylobacter isolates were subtyped by sequencing the short variable region of the flaA gene . The same subtypes of Campylobacter were detected in whole carcass rinse samples as in respiratory tract wash samples from individual broilers . Furthermore, the same numbers and subtypes of Campylobacter were recovered from respiratory tracts of carcasses collected before scalding and those collected after scalding . However, respiratory tracts of carcasses after scalding had higher numbers of E . coli, coliforms, and total aerobic bacteria than those tested before scalding . Although some bacterial counts were higher in the respiratory tracts of carcasses after scalding, Campylobacter counts were not . It appears that Campylobacter is present in the respiratory tracts of broilers as they enter processing, and contamination may be due to airborne bacteria during production or transport. J Food Prot, 2004 Jan, 67(1), 64 - 70 Development of predictive models for the survival of Campylobacter jejuni (ATCC 43051) on cooked chicken breast patties and in broth as a function of temperature; Yoon KS et al.; The objective of this study was to model the kinetics of the survival of Campylobacter jejuni on cooked chicken breast patties and in broth as a function of temperature . Both patties and broth were inoculated with 10(6) stationary-phase cells of a single strain of C . jejuni (ATCC 43051) and incubated at constant temperatures from 4 to 30 degrees C in 2 degrees C increments under aerobic conditions . In most cases, a three-phase linear model fit the primary survival curves well (r2 = 0.97 to 0.99) at all incubation temperatures regardless of model medium, indicating the presence of a resistant subpopulation of C . jejuni that would not be eliminated without thermal processing . Secondary models predicting lag time (LT) and specific death rate (SDR) as functions of temperature were also developed . The Davey and Boltzmann models were identified as appropriate secondary models for LT and SDR, respectively, on the basis of goodness of fit (Boltzmann model, r2 = 0.96; Davey model, r2 = 0.93) and prediction bias and accuracy factor tests . The results obtained indicate that C . jejuni can survive well at both refrigeration and ambient temperatures regardless of model medium . Reduced survival of C . jejuni, characterized by shorter lag times and faster death rates, was observed both on patties and in broth at ambient temperatures . In addition, the average maximum reduction of C . jejuni at 4 to 30 degrees C was 1.5 log units regardless of storage temperature or model medium . These findings suggest that C . jejuni found on contaminated poultry products has the potential to survive under conditions that are not permissive for growth and thus could cause foodborne illness if the poultry is not sufficiently cooked. Zh Mikrobiol Epidemiol Immunobiol, 2003 Nov-Dec, (6), 86 - 8 {Experience in the use of a new test system for the rapid indication of the causative agents of campylobacter infection under the conditions of an emergency situation}; Alieva EV; A new test system on the basis of magnetoimmunosorbents for the rapid indication of capmpylobacteriosis has been developed . Information on its use under the conditions of emergency situations is presented . Good prospects of this new test system in the study of environmental objects have been demonstrated. J Clin Microbiol, 2004 Jan, 42(1), 311 - 9 Serotypes, virulence genes, and intimin types of Shiga toxin (verotoxin)-producing Escherichia coli isolates from human patients: prevalence in Lugo, Spain, from 1992 through 1999; Blanco JE et al.; We have analyzed the prevalence of Shiga toxin-producing Escherichia coli (STEC) in stool specimens of patients with diarrhea or other gastrointestinal alterations from the Xeral-Calde Hospital of Lugo City (Spain) . STEC strains were detected in 126 (2.5%) of 5,054 cases investigated, with a progressive increase in the incidence from 0% in 1992 to 4.4% in 1999 . STEC O157:H7 was isolated in 24 cases (0.5%), whereas non-O157 STEC strains were isolated from 87 patients (1.7%) . STEC strains were (after Salmonella and Campylobacter strains) the third most frequently recovered enteropathogenic bacteria . A total of 126 human STEC isolates were characterized in this study . PCR showed that 43 (34%) isolates carried stx(1) genes, 45 (36%) possessed stx(2) genes and 38 (30%) carried both stx(1) and stx(2) . A total of 88 (70%) isolates carried an ehxA enterohemolysin gene, and 70 (56%) isolates possessed an eae intimin gene (27 isolates with type gamma1, 20 with type beta1, 8 with type zeta, 5 with type gamma2, and 3 with type epsilon) . STEC isolates belonged to 41 O serogroups and 66 O:H serotypes, including 21 serotypes associated with hemolytic uremic syndrome and 30 new serotypes not previously reported among human STEC strains in other studies . Although the 126 STEC isolates belonged to 81 different seropathotypes (associations between serotypes and virulence genes), only four accounted for 31% of isolates . Seropathotype O157:H7 stx(1) stx(2) eae-gamma1 ehxA was the most common (13 isolates) followed by O157:H7 stx(2) eae-gamma1 ehxA (11 isolates), O26:H11 stx(1) eae-beta1 ehxA (11 isolates), and O111:H- stx(1) stx(2) eae-gamma2 ehxA (4 isolates) . Our results suggest that STEC strains are a significant cause of human infections in Spain and confirm that in continental Europe, infections caused by STEC non-O157 strains are more common than those caused by O157:H7 isolates . The high prevalence of STEC strains (both O157:H7 and non-O157 strains) in human patients, and their association with serious complications, strongly supports the utilization of protocols for detection of all serotypes of STEC in Spanish clinical microbiology laboratories. J Clin Microbiol, 2004 Jan, 42(1), 229 - 35 Fluorescent amplified fragment length polymorphism genotyping of Campylobacter jejuni and Campylobacter coli strains and its relationship with host specificity, serotyping, and phage typing; Hopkins KL et al.; Fluorescent amplified fragment length polymorphism (FAFLP) analysis was applied to 276 Campylobacter jejuni strains and 87 Campylobacter coli strains isolated from humans, pigs, cattle, poultry, and retail meats to investigate whether certain FAFLP genotypes of C . jejuni and C . coli are associated with a particular host and to determine the degree of association between FAFLP-defined genotypes and heat-stable serotypes and/or phage types . Within C . coli, the poultry strains clustered separately from those of porcine origin . In contrast, no evidence of host specificity was detected among C . jejuni strains . While C . coli strains show host specificity by FAFLP genotyping, C . jejuni strains that are genotypically similar appear to colonize a range of hosts, rather than being host adapted . Some serotypes and/or phage types (C . jejuni serotype HS18, phage type PT6, and serophage type HS19/PT2 and C . coli HS66, PT2, and HS56/PT2) were the most homogeneous by FAFLP genotyping, while others were more heterogeneous (C . jejuni HS5 and PT39, and C . coli HS24 and PT44) and therefore poor indicators of genetic relatedness between strains . The lack of host specificity in C . jejuni suggests that tracing the source of infection during epidemiological investigations will continue to be difficult . The lack of congruence between some serotypes and/or phage types and FAFLP genotype underlines the need for phenotypic testing to be supplemented by genotyping . This study also demonstrates how, in general, FAFLP generates "anonymous" genetic markers for strain characterization and epidemiological investigation of Campylobacter in the food chain. Arch Microbiol, 2004 Feb, 181(2), 89 - 96 Epub 2004 Jan 09. Genomics of the ccoNOQP-encoded cbb3 oxidase complex in bacteria; Cosseau C et al.; Many bacteria adapt to microoxic conditions by synthesizing a particular cytochrome c oxidase (cbb3) complex with a high affinity for O2, encoded by the ccoNOQP operon . A survey of genome databases indicates that ccoNOQP sequences are widespread in all sub-branches of Proteobacteria but otherwise are found only in bacteria of the CFB group ( Cytophaga, Flexibacter, Bacteroides) . Our analysis of available genome sequences suggests four major strategies of regulating ccoNOQP expression in response to O2 . The most widespread strategy involves direct regulation by the O2-responsive protein Fnr . The second strategy involves an O2-insensitive paralogue of Fnr, FixK, whose expression is regulated by the O2-responding FixLJ two-component system . A third strategy of mixed regulation operates in bacteria carrying both fnr and fixLJ-fixKgenes . Another, not yet identified, strategy is likely to operate in the epsilon-Proteobacteria Helicobacter pylori and Campylobacter jejuni which lack fnr and fixLJ-fixK genes . The FixLJ strategy appears specific for the alpha-subclass of Proteobacteria but is not restricted to rhizobia in which it was originally discovered. Tidsskr Nor Laegeforen, 2003 Dec 4, 123(23), 3410 - 3 {Water-borne disease outbreaks in Norway}; Nygard K et al.; BACKGROUND: The drinking water in Norway has traditionally been considered being of good quality . However, outbreaks related to drinking water are reported every year . We review waterborne outbreaks in Norway over the last 15 years, and describe the aetiology of and contributory factors in these outbreaks . MATERIALS AND METHODS: We compiled data on waterborne outbreaks reported to the Norwegian Institute of Public Health and Norwegian Food Control Authority during 1988-2002 . We included all events in which two or more people fell ill and water was the suspected source of infection . RESULTS: Over the 15-year period . 72 outbreaks were reported, affecting a total of 10 616 persons . Campylobacter was the cause in 26% (19/72) of the outbreaks, norovirus in 18% (13/72) . The causative organism was unknown in 46% (33/72) . The water came from public waterworks in 32 of the 54 outbreaks for which this information was available (59%); from a private supply in the remaining 22 . For 62% (16/26) of the outbreaks related to waterworks, the water was not disinfected before distribution . None of the private water supplies were disinfected . Over the last five years, there were more outbreaks related to private supplies . INTERPRETATION: The most important contributory factor to waterborne outbreaks in Norway is contamination of the raw water combined with missing or faulty disinfecting procedures . To prevent future outbreaks, a continuous upgrading of small and private water supplies is needed . Reporting of outbreaks is important for the implementation of targeted and effective preventive measures. Appl Environ Microbiol, 2004 Jan, 70(1), 152 - 8 Detection of Salmonella enterica serovar typhimurium by using a rapid, array-based immunosensor; Taitt CR et al.; The multianalyte array biosensor (MAAB) is a rapid analysis instrument capable of detecting multiple analytes simultaneously . Rapid (15-min), single-analyte sandwich immunoassays were developed for the detection of Salmonella enterica serovar Typhimurium, with a detection limit of 8 x 10(4) CFU/ml; the limit of detection was improved 10-fold by lengthening the assay protocol to 1 h . S . enterica serovar Typhimurium was also detected in the following spiked foodstuffs, with minimal sample preparation: sausage, cantaloupe, whole liquid egg, alfalfa sprouts, and chicken carcass rinse . Cross-reactivity tests were performed with Escherichia coli and Campylobacter jejuni . To determine whether the MAAB has potential as a screening tool for the diagnosis of asymptomatic Salmonella infection of poultry, chicken excretal samples from a private, noncommercial farm and from university poultry facilities were tested . While the private farm excreta gave rise to signals significantly above the buffer blanks, none of the university samples tested positive for S . enterica serovar Typhimurium without spiking; dose-response curves of spiked excretal samples from university-raised poultry gave limits of detection of 8 x 10(3) CFU/g. Diagn Microbiol Infect Dis, 2003 Dec, 47(4), 601 - 8 Antimicrobial susceptibility trends in Campylobacter jejuni and Campylobacter coli isolated from a rural Egyptian pediatric population with diarrhea; Putnam SD et al.; 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 the linear trends over the duration of the study . There was no indication of frank macrolide (erythromycin or azithromycin) resistance among any Campylobacter . Moreover, there were statistically significant positive trends in both the MIC(50) and MIC(90) values for the erythromycin and azithromycin during the study period, suggesting a possible decreasing trend in susceptibility among Campylobacter . This study demonstrated that antimicrobial susceptibility in Campylobacter has significantly decreased from 1995 through 2000 among pediatric diarrhea cases in rural Egypt. Diagn Microbiol Infect Dis, 2003 Dec, 47(4), 551 - 6 Laribacter hongkongensis: a potential cause of infectious diarrhea; Woo PC et al.; In this study, we describe the isolation of Laribacter hongkongensis, a recently described genus and species of bacterium, in pure culture on charcoal cefoperazone deoxycholate agar from the stool of six patients with diarrhea . Three patients were residents of Hong Kong, and three of Switzerland . In none of the stool samples obtained from these six patients was Salmonella, Shigella, enterohemorrhagic Escherichia coli, Vibrio, Aeromonas, Plesiomonas, or Campylobacter recovered . Rotavirus antigen detection, electron microscopic examination for viruses, and microscopic examinations for ova and cysts were all negative for the stool samples obtained from the three patients in Hong Kong . Enterotoxigenic E . coli was recovered from one of the patients in Hong Kong . Unlike L . hongkongensis type strain HKU1, all the six strains were motile with bipolar flagellae . Sequencing of the 16S ribosomal RNA genes of the six strains showed that they all had sequences with only 0-2 base differences to that of the type strain . Pulsed field gel electrophoresis of the SpeI digested genomic DNA of the six isolates and that of the type strain revealed that the seven isolates were genotypically unrelated strains . More extensive epidemiologic studies should be carried out to ascertain the causative association between L . hongkongensis and diarrhea and to define the reservoir and modes of transmission of L . hongkongensis. Avian Dis, 2003 Oct-Dec, 47(4), 1429 - 33 Cecal colonization of chicks by bovine-derived strains of Campylobacter; Ziprin RL et al.; 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 from chick ceca were examined by ribotyping to assure they matched the challenge strain . One hundred percent of the bovine-derived challenge strains were capable of colonizing chicks . These results indicate that dairy cattle may be asymptomatic Campylobacter carriers and potential sources of campylobacteria contamination of poultry facilities. J Neurol Sci, 2004 Feb 15, 217(2), 225 - 8 Bickerstaff's brainstem encephalitis associated with IgM antibodies to GM1b and GalNAc-GD1a; Matsuo M et al.; This is the first report of a case of Bickerstaff's brainstem encephalitis (BBE) associated with IgM antibodies to GM1b and GalNAc-GD1a . Subsequent to Campylobacter jejuni enteritis, the patient rapidly developed consciousness disturbance and hyperreflexia in addition to external ophthalmoplegia and cerebellar-like ataxia . EEG showed transient 7 Hz monorhythmic theta activities, predominantly in the front-central area . He received high doses of immunoglobulin intravenously and had completely recovered 3 months later . High anti-GM1b and anti-GalNAc-GD1a IgM antibody titers present during the acute phase decreased with his clinical improvement . An absorption study showed the anti-GM1b and anti-GalNAc-GD1a IgM antibodies to be cross-reactive . Anti-GM1b and anti-GalNAc-GD1a antibodies have been detected in some patients who developed Guillain-Barre syndrome after C . jejuni enteritis, whereas the anti-GQ1b IgG antibody is associated with BBE . Infection by C . jejuni bearing a GM1b-like or GalNAc-GD1a-like lipooligosaccharide may trigger the production of anti-GalNAc-GD1a and anti-GM1b IgM antibodies . It is not clear why our patient developed BBE rather than Guillain-Barre syndrome . These antibodies may, however, prove useful serological markers for identifying BBE patients who do not have the anti-GQ1b IgG antibody. J Bacteriol, 2004 Jan, 186(2), 503 - 17 The genome-sequenced variant of Campylobacter jejuni NCTC 11168 and the original clonal clinical isolate differ markedly in colonization, gene expression, and virulence-associated phenotypes; Gaynor EC et al.; The genome sequence of the enteric bacterial pathogen Campylobacter jejuni NCTC 11168 (11168-GS) was published in 2000, providing a valuable resource for the identification of C . jejuni-specific colonization and virulence factors . Surprisingly, the 11168-GS clone was subsequently found to colonize 1-day-old chicks following oral challenge very poorly compared to other strains . In contrast, we have found that the original clinical isolate from which 11168-GS was derived, 11168-O, is an excellent colonizer of chicks . Other marked phenotypic differences were also identified: 11168-O invaded and translocated through tissue culture cells far more efficiently and rapidly than 11168-GS, was significantly more motile, and displayed a different morphology . Serotyping, multiple high-resolution molecular genotyping procedures, and subtractive hybridization did not yield observable genetic differences between the variants, suggesting that they are clonal . However, microarray transcriptional profiling of these strains under microaerobic and severely oxygen-limited conditions revealed dramatic expression differences for several gene families . Many of the differences were in respiration and metabolism genes and operons, suggesting that adaptation to different oxygen tensions may influence colonization potential . This correlates biologically with our observation that anaerobically priming 11168-GS or aerobically passaging 11168-O caused an increase or decrease, respectively, in colonization compared to the parent strain . Expression differences were also observed for several flagellar genes and other less well-characterized genes that may participate in motility . Targeted sequencing of the sigma factors revealed specific DNA differences undetected by the other genomic methods {corrected}. J Neuroimmunol, 2004 Jan, 146(1-2), 63 - 75 Induction of human IgM and IgG anti-GM1 antibodies in transgenic mice in response to lipopolysaccharides from Campylobacter jejuni; Lee G et al.; Campylobacter jejuni lipopolysaccharides (LPS) are implicated in the development of autoantibodies to GM1 ganglioside in patients with neuropathy following C . jejuni infection . CjLPS bears oligosaccharides that are cross reactive with GM1 ganglioside and presumably exerts its effects via molecular mimicry . To study the mechanisms that are involved in development of the autoantibody response, a transgenic mouse line was developed that expresses an IgM anti-GM1 antibody derived from a patient with multifocal motor neuropathy (MMN) . In vivo stimulation of the transgenic mice with C . jejuni lipopolysaccharides (CjLPS), but not of wild-type mice readily elicited high serum titers of anti-GM1 IgM antibodies, followed by IgG anti-GM1 antibodies after two booster injections . In in vitro experiments, CjLPS stimulated the transgenic B-cells at lower concentration than control LPS . The increased sensitivity to CjLPS and the induction of IgG anti-GM1 by CjLPS but not control LPS are consistent with a mechanism of B-cell activation that involves both the LPS and the antigen-specific surface Ig receptors, with possible participation of T-cells. J Infect Chemother, 2003 Dec, 9(4), 344 - 7 Isolation of Helicobacter cinaedi from blood of an immunocompromised patient in Japan; Murakami H et al.; We report the isolation of Helicobacter cinaedi (previously called " Campylobacter-like organism") from the blood of an immunosuppressed Japanese patient receiving immunosuppressive therapy after renal transplantation . The identification of H . cinaedi was based on the findings of microscopic examinations, biochemical properties, and 16S rRNA gene nucleotide sequences . H . cinaedi bacteremia should be considered as one of the critical infectious diseases in immunocompromised patients, and the sequencing of 16S rRNA may be a useful method to confirm the identification of this organism. Infect Immun, 2004 Jan, 72(1), 260 - 8 L-serine catabolism via an oxygen-labile L-serine dehydratase is essential for colonization of the avian gut by Campylobacter jejuni; Velayudhan J et al.; Campylobacter jejuni is a microaerophilic, asaccharolytic bacterium . The identity of the carbon and energy sources used by C . jejuni in vivo is unknown, but the genome sequence of strain NCTC11168 indicates the presence of genes for catabolism of a limited range of amino acids, including serine . Specific omission of L-serine from a defined medium containing a mixture of amino acids led to a dramatic decrease in cell yields . As C . jejuni does not have a biosynthetic serine requirement, this supports earlier suggestions that L-serine is a preferentially catabolized amino acid . Serine transport was found to be mediated by at least two systems in strain 11168; a high-capacity, low-affinity L-serine-specific system encoded by Cj1625c (sdaC) and a higher-affinity L-serine/L-threonine system responsible for residual L-serine transport in an sdaC mutant . Catabolism of L-serine to pyruvate and ammonia is carried out by SdaA (encoded by Cj1624c), which was overexpressed, purified, and shown to be an oxygen-labile iron-sulfur enzyme . L-Serine dehydratase activity in an sdaA mutant was reduced 10-fold compared to that in the wild type, but the residual activity (due to the anabolic L-threonine dehydratase) could not support either growth on or utilization of L-serine in defined media . However, although sdaA mutants showed no obvious growth defect in complex media, they completely failed to colonize 3-week-old chickens as assayed both by cloacal swabs taken over a 6-week period and by cecal colony counts postmortem . In contrast, the isogenic parent strain colonized chickens to high levels within 1 week of inoculation . The results show that an active SdaA is essential for colonization of the avian gut by C . jejuni and imply that catabolism of L-serine is crucially important for the growth of this bacterium in vivo. Lett Appl Microbiol, 2004, 38(1), 66 - 71 Chicken juice, a food-based model system suitable to study survival of Campylobacter jejuni; Birk T et al.; AIMS: The purpose of this study was to develop a food-based model system that resembles the environment that Campylobacter jejuni experiences on raw poultry products and use this model system to investigate growth and survival of the bacterium . METHODS AND RESULTS: Chicken juice was collected from frozen chickens and subsequently cleared by centrifugation and subjected to sterile filtration . At low temperatures (5 and 10 degrees C) C . jejuni NCTC11168 remained viable in chicken juice for a remarkably longer period of time than in the reference medium BHI . When exposed to heat stress (48 degrees C) C . jejuni NCTC11168 also showed increased viability in chicken juice compared with the reference medium . Furthermore, agar plates made with chicken juice supported growth of four clinical isolates of C . jejuni and a C . jejuni strain obtained from chicken at both 37 and 42 degrees C . CONCLUSIONS: Our work shows that minimal processed and sterilized chicken juice is an ideal environment for survival of C . jejuni and that it is useful as a food-based model system . SIGNIFICANCE AND IMPACT OF THE STUDY: The developed model system may contribute to the understanding of C . jejuni viability on poultry products and can be instrumental in the developme