Infect Immun, 1999 Aug, 67(8), 4171 - 82 Campylobacter jejuni 81-176 associates with microtubules and dynein during invasion of human intestinal cells; Hu L et al.; Campylobacter jejuni uptake into cultured INT407 cells was analyzed kinetically over a wide range of starting multiplicities of infection (MOI; from 0.02 to 20,000 bacteria/epithelial cell) . The efficiency of internalization was the highest at MOI of 0.02 and decreased steadily at higher MOIs, presumably due to reported C . jejuni autoagglutination at higher densities . Total internalized Campylobacter CFU increased gradually from an MOI of 0.02 to a peak at an MOI of 200 (reaching an average of two bacteria internalized per epithelial cell) and decreased at higher MOIs . The invasion process was apparently saturated within 2 h at an MOI of 200, indicating stringent host cell limitations on this entry process . Furthermore, whereas control Salmonella typhi invaded all monolayer cells within 1 h, only two-thirds of monolayer cells were infected after 2 h with C . jejuni at MOIs of 200 to 2,000 . The percentage of Campylobacter-infected host cells gradually increased to 85% after 7 h of infection, suggesting that C . jejuni entry may be host cell cycle dependent . Direct evidence of the involvement of microtubules in C . jejuni internalization, suggested previously by biochemical inhibitor studies, was obtained by time course immunofluorescence microscopic analyses . Bacteria initially bound to the tips of host cell membrane extensions containing microtubules, then aligned in parallel with microtubules during entry, colocalized specifically with microtubules and dynein but not with microfilaments, and moved over 4 h, presumably via microtubules to the perinuclear region of host cells . Orthovanadate, which inhibits dynein activity, specifically reduced C . jejuni 81-176 entry, suggesting that this molecular motor is involved in entry and endosome trafficking during this novel bacterial internalization process . Collectively, these data suggest that C . jejuni enters host cells in a targeted and tightly controlled process leading to uptake into an endosomal vacuole which apparently moves intracellularly along microtubules via the molecular motor, dynein, to the perinuclear region. Zentralbl Veterinarmed B, 1999 Jun, 46(5), 311 - 21 Health status of bulls used for natural breeding on farms in south west Scotland; McGowan AC et al.; One hundred and nine breeding bulls were examined during the period November 1992 to June 1993 on farms in south west Scotland for evidence of infectious diseases associated with breeding . Preputial washings were collected to screen for Campylobacter fetus venerialis, together with serial blood samples to assess their seroprevalence to Bovine Virus Diarrhoea virus (BVDv), Bovine Herpes Virus-1 (BHV-1), Leptospira hardjo and Bovine Herpes Virus-4 (BHV-4) . The possible impact of natural mating on the epidemiology of these diseases is described . Evidence of infections with Campylobacter fetus and BVH-4 were not found in this sample . The overall seroprevalence to BVDv was 78%, for BHV-1 49%, and L . hardjo 27% at titres of > or = 1/400 . This study shows that bulls may be responsible for the introduction and dissemination of these diseases when moved from farm to farm as part of normal cattle breeding in this area . Young unproven bulls may be particularly susceptible to endemic diseases associated with lowered reproductive performance. Am Fam Physician, 1999 Jul, 60(1), 119 - 24, 135-6 Prevention and treatment of traveler's diarrhea; Juckett G; Common pathogens in traveler's diarrhea include enterotoxigenic Escherichia coli, Campylobacter, Shigella, Salmonella, Yersinia and many other species . Viruses and protozoa are the cause in many cases . Fortunately, traveler's diarrhea can usually be avoided by carefully selecting foods and beverages . Although drug prophylaxis is now discouraged, treatment with loperamide (in the absence of dysentery) and a fluoroquinolone, such as ciprofloxacin (500 mg twice daily for one to three days), is usually safe and effective in adults with traveler's diarrhea . Trimethoprim-sulfamethoxazole and doxycycline are alternatives, but resistance increasingly limits their usefulness . Antibiotic treatment is best reserved for cases that fail to quickly respond to loperamide . Antibiotic resistance is now widespread . Nonabsorbable antibiotics, immunoprophylaxis with vaccines and biotherapeutic microbes that inhibit pathogen infection may eventually supplant antibiotic treatment . In the meantime, azithromycin and new fluoroquinolones show promise as possible replacements for the older agents . Ultimately, the best solution is improvements in sanitary engineering and the development of safe water supplies. J Neurol Neurosurg Psychiatry, 1999 Aug, 67(2), 180 - 4 Hyperreflexia in Guillain-Barré syndrome: relation with acute motor axonal neuropathy and anti-GM1 antibody; Kuwabara S et al.; OBJECTIVES: To investigate the incidence of hyperreflexia in patients with Guillain-Barre syndrome (GBS), and its relation with electrodiagnosis of acute motor axonal neuropathy (AMAN), antiganglioside GM1 antibody, and Campylobacter jejuni infection . It was reported that patients with AMAN in northern China often had hyperreflexia in the recovery phase . METHODS: In 54 consecutive Japanese patients with GBS, sequential findings of tendon reflexes were reviewed . By electrodiagnostic criteria, patients were classified as having AMAN or acute inflammatory demyelinating polyneuropathy (AIDP) . Anti-GM1 and anti-C jejuni antibodies were measured by enzyme linked immunosorbent assays . RESULTS: Seven (13%) patients developed hyperreflexia with the spread of the myotatic reflex to other segments in the early recovery phase, one of whom already had hyperreflexia in the acute progressive phase . Of the seven patients, six had AMAN and all seven had anti-GM1 antibodies, whereas only two had anti-C jejuni antibodies . Hyperreflexia was more often found in patients with AMAN than AIDP (6/23 v 1/18, p=0 . 002), and in patients with anti-GM1 antibodies than without them (7/26 v 0/28, p=0.01) . Hyperreflexic patients had milder peak disabilities than patients without hyperreflexia (p=0.03) . Increased motor neuron excitability in the hyperreflexic patients was supported by increased soleus H-reflex amplitudes and the appearance of H-reflexes in the small hand or foot muscles . CONCLUSIONS: Hyperreflexia often occurs in patients with GBS especially with AMAN, anti-GM1 antibodies, and milder disease . Increased motor neuron excitability further characterises the subgroup of patients with GBS with AMAN and anti-GM1 antibodies. J Chromatogr B Biomed Sci Appl, 1999 May 28, 728(2), 273 - 7 High-performance liquid chromatography determination of residue levels on chicken carcasses treated with cetylpyridinium chloride; Zhou X et al.; Cetylpyridinium chloride (CPC) has been found to be effective in reducing contamination of chicken carcasses from a variety of microorganisms, including Escherichia coli O157:H7, Salmonella typhimurium, Campylobacter jejuni, Aeromonas hydrophila, Listeria monocytogenes, and Staphylococcus aureus . A procedure has been developed to determine residue levels on chicken carcasses after CPC treatment . For the analysis, chicken carcasses were extracted with 95% ethanol . The CPC concentration in the extract was measured by high-performance liquid chromatography (HPLC) with ultraviolet detection using dodecylpyridinium chloride (DPC) as an internal standard . The method was validated in the concentration range of 3-200 microg/ml CPC in ethanolic extract . This assay is rapid, precise, and accurate. J Clin Microbiol, 1999 Aug, 37(8), 2656 - 62 Disseminated zygomycosis due to Rhizopus schipperae after heatstroke; Anstead GM et al.; A 21-year-old woman suffered heatstroke and developed diarrhea while trekking across south Texas . The heatstroke was complicated by seizures, rhabdomyolysis, pneumonia, renal failure, and disseminated intravascular coagulation . The patient's stool and blood cultures grew Campylobacter jejuni . The patient subsequently developed paranasal and gastrointestinal zygomycosis and required surgical debridement and a prolonged course of amphotericin B . The zygomycete cultured was Rhizopus schipperae . This is only the second isolate of R . schipperae that has been described . R . schipperae is characterized by the production of clusters of up to 10 sporangiophores arising from simple but well-developed rhizoids . These asexual reproductive propagules are produced on Czapek Dox agar but are absent on routine mycology media, where only chlamydospores are observed . Despite multiorgan failure, bacteremia, and disseminated zygomycosis, the patient survived and had a good neurological outcome . Heatstroke has not been previously described as a risk factor for the development of disseminated zygomycosis. J Med Microbiol, 1999 Jul, 48(7), 617 - 21 Comparison of a commercial test for serotyping heat-stable antigens of Campylobacter jejuni with genotyping by pulsed-field gel electrophoresis; Rautelin H et al.; A new commercial serotyping set based on heat-stable Penner's antigens was compared with pulsed-field gel electrophoresis (PFGE) with SmaI and SacII restriction endonucleases . Among 50 isolates of Campylobacter jejuni from Finnish patients, which represented predominant PFGE patterns selected from isolates from sporadic cases and isolates associated with small outbreaks, 11 different serotypes were demonstrated from 43 typable isolates . Several PFGE patterns could be found within one serotype; on the other hand, several serotypes could be demonstrated within one PFGE type . Most isolates originated from sporadic cases; however, some isolates were epidemiologically associated and showed identical serotypes and PFGE patterns . Although the new serotyping set would have been useful in the few epidemic cases studied, several isolates (14%) representing the major PFGE patterns remained untypable or gave weakly positive agglutination reactions only suggesting a plausible serotype (18%) . This might restrict the use of the novel serotyping set, at least in Finland. Arthritis Rheum, 1999 Jul, 42(7), 1386 - 96 No benefit of long-term ciprofloxacin treatment in patients with reactive arthritis and undifferentiated oligoarthritis: a three-month, multicenter, double-blind, randomized, placebo-controlled study; Sieper J et al.; OBJECTIVE: To investigate the effect of long-term antibiotic treatment in patients with reactive arthritis (ReA) and undifferentiated oligoarthritis . METHODS: One hundred twenty-six patients were treated with ciprofloxacin (500 mg twice a day) or placebo for 3 months, in a double-blind, randomized study . Of these patients, 104 (48 treated with ciprofloxacin and 56 treated with placebo) were valid for clinical evaluation: 55 were diagnosed as having ReA with a preceding symptomatic urogenic or enteric infection and 49 as having undifferentiated oligoarthritis . These 2 groups were randomized separately . The triggering bacterium was sought by serology and/or culture . The percentage of patients in remission after 3 months of treatment was chosen as the primary efficacy parameter . RESULTS: A triggering bacterium could be identified in 52 patients (50%): Chlamydia trachomatis in 13, Yersinia in 14, and Salmonella in 25 . No patient was positive for Campylobacter jejuni or for Shigella . No difference in outcome was found between treatment with ciprofloxacin or placebo in the whole group or in subgroups of patients with ReA or undifferentiated oligoarthritis . No difference was seen in patients with a disease duration <3 months . Ciprofloxacin was not effective in Yersinia- or Salmonella-induced arthritis but seemed to be better than placebo in Chlamydia-induced arthritis . This difference was not significant, however, which might be due to the small sample size . CONCLUSION: Long-term treatment of ReA with ciprofloxacin is not effective; however, it might be useful in the subgroup of patients who have Chlamydia-induced arthritis . This has to be proven in a bigger study focusing on patients with Chlamydia-induced arthritis. Commun Dis Public Health, 1999 Jun, 2(2), 114 - 8 Microbiological investigation of halal butchery products and butchers' premises; Little C et al.; Halal butchers' premises were investigated as they had not been represented in a recent study of butchery products and butchers' premises conducted by the Local Authorities Coordinating Body on Food and Trading Standards and the PHLS . This study examined 183 raw prepared meats and 212 environmental samples from 105 halal butcher premises . Only raw meats were prepared on 97 of the premises visited; and the types of meat prepared on the remaining eight premises was not specified . Four halal butchers sold cooked meats prepared elsewhere . Salmonella spp . and Campylobacter spp . were detected in 12 (7%) and 52 (28%) of the 183 raw meat products, respectively . Five raw prepared meats (3%) contained both salmonella and campylobacter . Vero cytotoxin producing Escherichia coli O157 was isolated from a raw meat product that also contained campylobacter . No cooked meat products were available for collection . The physical separation of raw and unwrapped cooked meat products in premises that prepared raw and sold cooked meats was not recorded . Apron cloths were the most heavily contaminated environmental samples examined; hygiene indicator microorganisms indicated an increased risk of cross contamination . Managers in 85 premises had received no food hygiene training and 88 premises had no hazard analysis system in place . Improvements are needed to reduce the risk of cross contamination. Commun Dis Public Health, 1999 Jun, 2(2), 108 - 13 A study of infectious intestinal disease in England: microbiological findings in cases and controls; Tompkins DS et al.; A study was undertaken to identify the microorganisms and toxins in stool specimens associated with infectious intestinal disease (IID) among cases in the community and presenting to general practitioners (GPs) and in asymptomatic controls . Population based cohorts were recruited from practice lists in 70 practices and followed for 26 weeks (cohort component) . Seven hundred and sixty-one cases of IID identified from the cohorts, 2893 cases who presented to GPs in 34 of the practices (GP component), and age/sex matched control subjects (555 and 2264, respectively) submitted stool specimens by post for comprehensive microbiological examination . Campylobacter spp (12.2% of stools tested), rotavirus group A (7.7%), and small round structured virus (SRSV) (6.5%) were the organisms most commonly detected in the GP component . SRSV was identified in 7.0% of cases in the community cohort . No target microorganisms or toxins were identified in 45.1% and 63.1% of cases in the two components . Aeromonas spp, Yersinia spp, and some enterovirulent groups of Escherichia coli were detected as frequently in controls as in cases . The higher frequency of detection of campylobacter, salmonella, and rotavirus among cases who presented to GPs than among those in the community suggests that those pathogens cause more severe illness . No enteropathogens were detected from a large proportion of cases although comprehensive standard methods were used to seek them. J Neurol Sci, 1999 Apr 1, 164(2), 134 - 8 Autoantibodies to GM1b and GalNAc-GD1a: relationship to Campylobacter jejuni infection and acute motor axonal neuropathy in China; Yuki N et al.; IgG antibodies to the minor gangliosides GM1b and GalNAc-GD1a frequently are present in sera of Japanese patients with Guillain-Barre syndrome . The relationship between these autoantibodies and Campylobacter jejuni infection, the type of disease (acute motor axonal neuropathy {AMAN}, or acute inflammatory demyelinating polyneuropathy {AIDP}) has yet to be established . Sera samples were obtained from 55 Chinese patients with clinically defined Guillain-Barre syndrome . An electrophysiology study showed nine AIDP, 28 had AMAN, and 18 unclassified . C . jejuni serology was positively correlated with anti-GM1b and anti-GalNAc-GD1a IgG antibodies (respective P values, 0.007 and 0.02) . The frequencies of positive anti-GM1b and anti-GalNAc-GD1a serology were greater in AMAN (32 and 21%) than in AIDP (11 and 0%), but the differences were not significant . Infection by C . jejuni may induce IgG anti-GM1b antibody in some patients and IgG anti-GalNAc-GD1a antibody in others . A larger population of patients must be studied to show whether there is a definite correlation. Avian Dis, 1999 Apr-Jun, 43(2), 245 - 50 Inhibition of Salmonella typhimurium in the chicken intestinal tract by a transformed avirulent avian Escherichia coli; Wooley RE et al.; An avirulent, wild-type avian Escherichia coli (E . coli Av) was electrotransformed with a plasmid coding for the production of microcin 24 (pGOB18) and was designated E . coli AvGOB18 . The transformant inhibited the growth of seven serotypes of Salmonella commonly associated with colonization and contamination of poultry products and seven strains of E . coli O157:H7 in the in vitro colicin/microcin assay . The transformant did not inhibit the replication of multiple isolates of Listeria monocytogenes or Campylobacter jejuni in similar assays . The transformant is nonconjugative, indicating that the plasmid would not be transmitted to other intestinal microflora in the environment . The transformant also survived in sterile tap and deionized water incubated at 25 C and 37 C in the laboratory for 30 days and was recovered from drinkers and birds in in vivo floor pen studies . In in vivo studies, E . coli AvGOB18 did not colonize the intestinal tract of broiler chicks when given as a single or multiple dose and did not reduce the Salmonella load in the broilers . But Salmonella typhimurium was reduced significantly in the intestinal tracts of broiler chickens when E . coli AvGOB18 was administered continually in the water supply. FEMS Immunol Med Microbiol, 1999 Jun, 24(2), 189 - 92 Susceptibility in vitro of Helicobacter pylori to cetylpyridinium chloride; Bereswill S et al.; The antimicrobial agent cetylpyridinium chloride (CPC) which is used in therapy of oro-pharyngeal infections and for antiseptic treatment of the oral cavity is active against different bacterial species . Determination of the minimal inhibitory concentration (MIC) using the agar dilution technique revealed that the gastric pathogen Helicobacter pylori in vitro is highly susceptible to CPC as indicated by an MIC of 10 microM (3.4 microg ml(-1)) which was significantly lower than the MIC of CPC against other bacterial species, which were analyzed in comparison to H . pylori . Bacteria of the genus Campylobacter, various Streptococcus spp., Staphylococcus aureus and Escherichia coli showed higher MICs ranging from 100 microM to 2 mM . In summary, this finding renders CPC-containing drugs candidates possibly useful for eradication or for the prevention of transmission of the gastric pathogen. Rinsho Shinkeigaku, 1999 Jan, 39(1), 17 - 8 {Campylobacter jejuni enteritis and Guillain-Barré syndrome}; Yuki N; Guillain-Barre syndrome (GBS) is the most common cause of acute neuromuscular paralysis . Sera from patients with GBS following Campylobacter jejuni infection frequently have autoantibody to GM 1 ganglioside in the acute phase of the illness . We revealed that the lipopolysaccharide (LPS) of C . jejuni that was isolated from a GBS patient has the oligosaccharide structure {Gal beta 1-3 GalNAc beta 1-4 (NeuAc alpha 2-3) Gal beta 1-}, which is identical to the terminal tetrasaccharide of GM 1 ganglioside . (1) Infection by C . jejuni that bears the GM 1-like lipopolysaccharide associated with the serotypic determinant of PEN 19 induces high production of IgG 1 and IgG 3 anti-GM 1 antibodies with help of T cells . (2) IgG anti-GM 1 antibody binds to motor nerve terminal axons, inhibits motoneuron excitability, and produces the development of GBS. Infect Immun, 1999 Jul, 67(7), 3698 - 701 Human monoclonal immunoglobulin M antibodies to ganglioside GM1 show diverse cross-reactivities with lipopolysaccharides of Campylobacter jejuni strains associated with Guillain-Barré syndrome; Prendergast MM et al.; We examined the reactivity of a panel of anti-GM1 immunoglobulin M monoclonal antibodies (MAbs) cloned from multifocal motor neuropathy patients with lipopolysaccharides (LPSs) of Campylobacter jejuni strains, including serotype O:41 strains associated with Guillain-Barre syndrome . The MAbs reacted with ganglioside GM1 to different degrees, and these differences in fine specificities for GM1 were reflected in the different degrees of reactivity with each of the C . jejuni LPSs tested . Antibodies could also be discriminated by the varying patterns of inhibition by cholera toxin (a GM1 ligand) in LPS binding studies . These results indicate that there is a substantial heterogeneity among C . jejuni O:41 strains in their expression of GM1-like epitopes and among the fine specificities of different neuropathy-associated anti-GM1 antibodies. Res Microbiol, 1999 May, 150(4), 247 - 55 Campylobacter coli strains with enlarged flagellin genes isolated from river water; Linton D et al.; A group of campylobacters isolated from river water were found to possess unusually large flagellin genes . Both phenotype and serology were consistent with identification as Campylobacter coli . Phylogenetic analysis of small (16S, rrs) and large subunit (23S, rrl) rRNA genes of a representative strain, NCTC 13006, demonstrated high levels of relatedness with C . jejuni and C . coli (99.1 and 98.3% similarity for 16S; 99.3 and 99.4% similarity for 23S) . Large flagellin proteins were demonstrated by SDS-PAGE analysis . The flaA and flaB genes were sequenced and aligned with known campylobacter flagellin amino acid sequences . The encoded FlaA protein of the new group exhibited a high degree of divergence from other Campylobacter species . Within the central variable region of FlaA, a further hypervariable domain was identified containing characteristic repeated motifs . Separate pairwise alignments performed for the variable regions of the polypeptide indicated these large fla genes were more closely related to those of C . upsaliensis than to those of C . coli or C . jejuni. Rheumatology (Oxford), 1999 May, 38(5), 411 - 4 Chlamydia pneumoniae as a triggering infection in reactive arthritis; Hannu T et al.; OBJECTIVE: To determine the role of Chlamydia pneumoniae as a triggering infection in reactive arthritis (ReA) . METHODS: Sixty patients with acute arthritis were screened for the evidence of triggering infections . In all patients, bacterial stool cultures, culture of Chlamydia trachomatis in urethra/cervix, and/or bacterial serology were studied . Chlamydia pneumoniae antibodies were measured by specific microimmunofluorescence test . RESULTS: Thirty-five of 60 patients fulfilled the diagnostic criteria for ReA . Thirty-one patients had microbial/serological evidence of preceding infection due to Salmonella, Yersinia, Campylobacter or Chlamydia trachomatis, or they had enteritis or urethritis prior to arthritis . Four additional patients had high antibody titre for C . pneumoniae . Three of these four patients had preceding lower respiratory symptoms, and were positive for HLA-B27 . The clinical picture of C . pneumoniae-positive ReA patients was similar to that of ReA patients with other definite aetiology . CONCLUSION: Chlamydia pneumoniae is a triggering factor in approximately 10% of patients with acute ReA. J Bacteriol, 1999 Jun, 181(12), 3857 - 9 Fused and overlapping rpoB and rpoC genes in Helicobacters, Campylobacters, and related bacteria; Zakharova N et al.; The genes coding for the beta (rpoB) and beta' (rpoC) subunits of RNA polymerase are fused in the gastric pathogen Helicobacter pylori but separate in other taxonomic groups . To better understand how the unique fused structure evolved, we determined DNA sequences at and around the rpoB-rpoC junction in 10 gastric and nongastric species of Helicobacter and in members of the related genera Wolinella, Arcobacter, Sulfurospirillum, and Campylobacter . We found the fusion to be specific to Helicobacter and Wolinella genera; rpoB and rpoC overlap in the other genera . The fusion may have arisen by a frameshift mutation at the site of rpoB and rpoC overlap . Loss of good Shine-Dalgarno sequences might then have fixed the fusion in the Helicobacteraceae, even if fusion itself did not confer a selective advantage. J Clin Microbiol, 1999 Jul, 37(7), 2376 - 7 Evaluation of the AnaeroPack Campylo system for growth of microaerophilic bacteria; Van Horn K et al.; Growth of microaerophilic bacteria in the AnaeroPack Campylo (Mitsubishi Gas Chemical America, Inc., New York, N.Y.) atmosphere generation system was compared to growth in the CampyPak Plus jar and CampyPak pouch (Becton-Dickinson Microbiology Systems {BDMS}, Cockeysville, Md.) . Growth in the AnaeroPack Campylo system was considered equivalent to or better than growth obtained in the CampyPak Plus and CampyPak pouch systems for 48 of the 50 Helicobacter pylori strains and for all 28 Campylobacter species tested . All of the 78 organisms tested were recovered in each system in equivalent colony counts . Two strains of H . pylori grown in the AnaeroPack Campylo system were observed to have colony morphology growth discrepancies when compared to growth in the two BDMS systems . Atmosphere failure with the AnaeroPack Campylo was not detected with Campylobacter jejuni ATCC 33291 used as a growth control . The AnaeroPack Campylo system is easy to use and supports the growth of campylobacters and H . pylori. Mol Microbiol, 1999 May, 32(4), 691 - 701 Bacterial secreted proteins are required for the internaliztion of Campylobacter jejuni into cultured mammalian cells; Konkel ME et al.; Presented here is the first evidence that Campylobacter jejuni secrete proteins upon co-cultivation with host cells and in INT 407 cell-conditioned medium . A C . jejuni gene designated ciaB for Campylobacter invasion antigen B was identified, using a differential screening technique, which is required for this secretion process and the efficient entry of this bacterium into a host cell . The C . jejuni ciaB gene encodes a protein of 610 amino acids with a calculated molecular mass of 73 154 Da . The deduced amino acid sequence of the CiaB protein shares similarity with type III secreted proteins associated with the invasion of host cells from other more extensively characterized bacterial pathogens . In vitro binding and internalization assays revealed that the binding of C . jejuni ciaB null mutants was indistinguishable from that of the parental isolate, whereas a significant reduction was noted in internalization . Confocal microscopic examination of C . jejuni-infected cells revealed that CiaB was translocated into the cytoplasm of the host cells . Culturing C . jejuni with INT 407 cells or in INT 407-conditioned medium resulted in the secretion of at least eight proteins, ranging in size from 12.8 to 108 kDa, into the culture medium . C . jejuni ciaB null mutants were deficient in the secretion of all eight proteins, indicating that CiaB is required for the secretion process . The identification of the C . jejuni ciaB gene represents a significant advance in understanding the molecular mechanism of C . jejuni internalization and the pathogenesis of C . jejuni-mediated enteritis. Mol Microbiol, 1999 Jun, 32(5), 1022 - 30 Evidence for a system of general protein glycosylation in Campylobacter jejuni; Szymanski CM et al.; A genetic locus from Campylobacter jejuni 81-176 (O:23, 36) has been characterized that appears to be involved in glycosylation of multiple proteins, including flagellin . The lipopolysaccharide (LPS) core of Escherichia coli DH5alpha containing some of these genes is modified such that it becomes immunoreactive with O:23 and O:36 antisera and loses reactivity with the lectin wheat germ agglutinin (WGA) . Site-specific mutation of one of these genes in the E . coli host causes loss of O:23 and O:36 antibody reactivity and restores reactivity with WGA . However, site-specific mutation of each of the seven genes in 81-176 failed to show any detectable changes in LPS . Multiple proteins from various cellular fractions of each mutant showed altered reactivity by Western blot analyses using O:23 and O:36 antisera . The changes in protein antigenicity could be restored in one of the mutants by the presence of the corresponding wild-type allele in trans on a shuttle vector . Flagellin, which is known to be a glycoprotein, was one of the proteins that showed altered reactivity with O:23 and O:36 antiserum in the mutants . Chemical deglycosylation of protein fractions from the 81-176 wild type suggests that the other proteins with altered antigenicity in the mutants are also glycosylated. Curr Opin Biotechnol, 1999 Jun, 10(3), 273 - 8 Novel detection techniques for human pathogens that contaminate poultry; Mandrell RE et al.; Poultry products are presumed to be a major contributor to human foodborne illness due to their high frequency of contamination with pathogens Salmonella spp . and Campylobacter spp . This has stimulated the development of more sensitive and rapid methods for identifying pathogens present in poultry . These new methods include immunomagnetic separation of pathogen, PCR amplification of pathogen-specific sequences, pathogen-specific DNA and RNA probes, and identification of pathogen-specific ions by mass spectrometry. Ann Neurol, 1999 Jun, 45(6), 758 - 68 Anti-GalNAc-GD1a antibody-associated Guillain-Barré syndrome with a predominantly distal weakness without cranial nerve impairment and sensory disturbance; Hao Q et al.; The serum antibodies to N-acetylgalactosaminyl GD1a (GalNAc-GD1a) and other gangliosides as well as to Campylobacter jejuni were determined in 147 patients with Guillain-Barre syndrome (GBS) . We found a distinctive clinical pattern in patients with anti-GalNAc-GD1a antibodies compared with those without the antibodies, that is, lack of cranial nerve involvement (87% versus 38%), distal-dominant weakness (80% versus 25%), and no sensory disturbance (73% versus 22%) . The frequency of distal-dominant weakness was significantly higher in patients with both C . jejuni infection and anti-GalNAc-GD1a positivity (100%) than in C . jejuni-negative/anti-GalNAc-GD1a-positive (25%), C . jejuni-positive/anti-GalNAc-GD1a-negative (32%) and C . jejuni-negative/anti-GalNAc-GD1a-negative patients (20%) . Lack of cranial nerve involvement and sensory disturbance were found in most C . jejuni-positive/anti-GalNAc-GD1a-positive and C . jejuni-negative/anti-GalNAc-GD1a-positive patients, but not in C . jejuni-positive/anti-GalNAc-GD1a-negative and C . jejuni-negative/anti-GalNAc-GD1a-negative patients . Although the anti-GM1-positive/anti-GalNAc-GD1a-negative patients mostly (75%) lacked cranial nerve involvement, distal-dominant weakness (38%) and lack of sensory disturbance (13%) were infrequent . These results may indicate that (1) the combination of C . jejuni infection and anti-GalNAc-GD1a antibodies, but not anti-GalNAc-GD1a, anti-GM1, or C . jejuni infection alone, is associated with a predominantly distal weakness, (2) the presence of anti-GalNAc-GD1a, rather than C . jejuni infection or anti-GM1 antibody, is associated with a lack of sensory disturbance, (3) both anti-GalNAc-GD1a and anti-GM1 antibodies are independently associated with a lack of cranial nerve impairment. J Med Microbiol, 1999 Jun, 48(6), 601 - 3 Fatal Campylobacter jejuni bacteraemia in patients with AIDS; Manfredi R et al.; Two fatal cases of Campylobacter jejuni septicaemia in patients with AIDS were characterised by severe HIV-related immunodeficiency, negative stool cultures and presentation during hospitalisation, developing a clinical picture of fulminant septic shock despite therapy with appropriate antibiotics . Campylobacter spp . are important opportunist pathogens in HIV disease and may cause a septicaemic illness in the absence of enteric disease. J Med Microbiol, 1999 Jun, 48(6), 523 - 6 Significance of Cryptosporidium in acute diarrhoea in North-Eastern India; Nath G et al.; In a hospital-based study, stool samples from 2095 patients of all ages were examined for different fungal, protozoal and bacterial enteropathogens over a period of 2 years (July 1994-June 1996) . Cryptosporidium was detected in 151 specimens (7.2%) and was the third commonest pathogen found . The highest prevalence of this organism was in the group aged 16-45 years and during the rainy months (July-Oct.) . Diarrhoea caused by the protozoon was of mild to moderate severity and features of dysentery were absent . Amongst other enteropathogens, Candida albicans was the most frequently isolated, followed by enteropathogenic and enterotoxigenic Escherichia coli, Salmonella spp., Campylobacter jejuni, Entamoeba histolytica, Giardia duodenalis (lamblia), Shigella spp., Vibrio cholerae and Aeromonas spp. Microbiol Res, 1999 May, 154(1), 23 - 6 Identification of bacteria using two degenerate 16S rDNA sequencing primers; Boye K et al.; Two degenerate 16S rDNA primers have been designed for broad-range identification of eubacteria by PCR and automated sequencing . Using a simple method, the primers have proven useful in identification of proteobacteria (Campylobacter, Enterobacter, Escherichia, Helicobacter, Klebsiella), gram-positive bacteria (Mycobacterium, Staphylococcus, Streptococcus) and spirochetes (Borrelia) derived from clinical samples . In several cases, the samples could be identified at the species level. Diagn Microbiol Infect Dis, 1999 Jun, 34(2), 99 - 102 Comparative antimicrobial activity of gatifloxacin tested against Campylobacter jejuni including fluoroquinolone-resistant clinical isolates; Hayward CL et al.; Campylobacter jejuni is an important pathogen that causes gastroenteritis, as well as other disease states such as meningitis and septic arthritis . In this study, the Etest (AB BIODISK, Solna, Sweden) results were compared to a reference agar dilution method using gatifloxacin, a new 8-methoxyfluoroquinolone . A total of 53 strains of C . jejuni initially isolated from patients in California and Mexico were tested . Results demonstrated a high correlation (r = 0.88) between the two utilized in vitro dilution methods . In addition, gatifloxacin activity was compared to that of ciprofloxacin, metronidazole, amoxicillin, erythromycin, chloramphenicol, gentamicin, tetracycline, and trimethoprim/sulfamethoxazole using the Etest . Gatifloxacin (MIC90, 4 micrograms/ml) was approximately eight- to 16-fold more potent than ciprofloxacin (Mic90, > 32 micrograms/ml), a commonly used fluoroquinolone for Campylobacter infections . Eight strains highly resistant to ciprofloxacin (MIC90, > 32 micrograms/ml) were tested for cross resistance against the newer fluoroquinolones (gatifloxacin, levofloxacin, trovafloxacin) and the rank order of potency was: gatifloxacin (MIC50, 16 micrograms/ml) > trovafloxacin = levofloxacin (MIC50, > 32 micrograms/mL) . However, only 25% ciprofloxacin-resistant strains were inhibited by < or = 1 microgram/mL of gatifloxacin or trovafloxacin . These results for gatifloxacin against C . jejuni strains must be further assessed in the context of in vivo trials before the clinical role of this new fluoroquinolone can be determined . The Etest appears to be a simple and precise susceptibility test method for testing C . jejuni isolates against fluoroquinolones and other alternative therapeutic agents. Pediatrics, 1999 Jun, 103(6 Pt 1), 1189 - 92 Impact of simple screening criteria on utilization of low-yield bacterial stool cultures in a Children's Hospital; Zaidi AK et al.; OBJECTIVE: To determine diagnostic yield of stool cultures for Salmonella, Shigella, Campylobacter jejuni, Yersinia enterocolitica, and Escherichia coli O157:H7 (SSCYE) among hospitalized children and to develop guidelines for appropriate use of these tests . Setting . Tertiary care pediatric hospital . DESIGN: Computerized records from the Microbiology Laboratory from January 1992 to December 1996 were reviewed retrospectively to collect data on the number of stool cultures performed in inpatients and outpatients, the length of hospital stay at the time cultures were sent, and diagnostic yield of cultures in hospitalized patients . A detailed review of medical records of all patients with a stool pathogen isolated after 3 days of hospitalization was also undertaken . The results from this retrospective analysis were used to develop guidelines to reduce unwarranted stool cultures and to educate medical care providers in the appropriate use of these tests . The impact of these guidelines on reduction in the volume of stool cultures performed on hospitalized patients was measured prospectively from January 1998 to June 1998 . RESULTS: A total of 27 110 stool cultures for SSCYE were performed in the 5-year study period . Of the 14 125 cultures from inpatients, 174 (1.2%) were positive . Among the cultures from inpatients, 9378 (66%) were from patients hospitalized for >3 days . Only 13 (.14%) were positive . Of these 13 cultures, 4 represented nosocomial infections, whereas the remaining 9 cultures either were sent to document clearance from a patient known previously to be infected with an enteric pathogen (7), or were attributed to delayed testing in individuals admitted with a diarrheal illness (2) . Introduction of guidelines to reject all SSCYE cultures from patients hospitalized for >3 days who did not meet specified criteria was associated with an overall reduction of 689 (43%) in the volume of tests performed in the 6-month period evaluated . This included 497 fewer cultures ordered and 192 cultures that were ordered but rejected because screening criteria were not met . Only 11 (5.4%) of 203 cultures sent >3 days after admission were processed because they met clinical criteria for testing . None were positive . Estimated cost savings were $50 163/year . CONCLUSIONS: Stool cultures for SSCYE among hospitalized patients have very low diagnostic yield and are extremely overutilized . Simple guidelines, such as rejecting (with few exceptions) cultures from patients hospitalized for >3 days, can reduce substantially such unnecessary testing. Can J Microbiol, 1999 Jan, 45(1), 23 - 30 Isolation and purification of a Campylobacter upsaliensis autolysin; Santiwatanakul S et al.; Autolytic activity in the soluble and sediment fractions of sonicates of the spiral and the coccoid form of Campylobacter upsaliensis could not be demonstrated by native (nondenaturing) polyacrylamide gel electrophoresis (PAGE) . Autolysins were detected, however, by using denaturing sodium dodecyl sulfate (SDS)-PAGE gels containing either purified Escherichia coli peptidoglycan or whole cells of Micrococcus luteus (Micrococcus lysodeikticus) as the turbid substrate, with subsequent renaturation by treatment with Triton X-100 buffer . In renaturing gels that contained Escherichia coli peptidoglycan, 14 putative autolytic bands ranging from 200 to 12 kDa were detected . In similar gels containing whole cells of M . luteus, only a single band appeared with a molecular mass of 34 kDa . This band corresponded to one of the bands present in the gels containing Escherichia coli peptidoglycan . This common autolysin was isolated by adsorbing it from Campylobacter upsaliensis soluble fractions onto M . luteus cells and then subjecting these cells to renaturing SDS-PAGE in gels containing Escherichia coli peptidoglycan . The 34-kDa autolysin differed from a single 51-kDa autolysin unique to the M . luteus cells, and when isolated from an SDS-PAGE gel, was pure when tested by isoelectric focusing . The N-terminal amino acid sequence analysis showed the first 15 amino acids of the 34-kDa autolysin to have 67% identity to a part of antigenic protein PEB4 of Campylobacter jejuni . The purified autolysin was used to immunize rabbits and the antibodies produced precipitated autolytic activity from cell lysates . The specificity of the antibodies was shown by Western blotting: only a single specific band occurred, with a molecular mass of 34 kDa, and thus it seems unlikely that the 34-kDa autolysin was derived from any of the other autolysins that were detected. J Med Liban, 1998 Nov-Dec, 46(6), 310 - 6 Prevalence, antimicrobial susceptibility and molecular characterization of Campylobacter isolates recovered from humans and poultry in Lebanon; Talhouk RS et al.; Recovery of Campylobacter was attempted from 281 consecutive non selected out-patients diarrheic stools, 150 individual ceca collected from meat chicken breeder farms and 31 slaughtered marketed chicken obtained from shops in Lebanon . Campylobacter isolates were recovered from 2 (0.7%) human stool specimens, 34 (22.7%) chicken ceca and 3 (9.7%) raw chicken carcasses . Speciation of these isolates revealed 2 C . jejuni from humans diarrheic stools, 16 C . coli, 10 C . jejuni, 3 C . fetus, 2 C . fennelliae (Helicobacter fennelliae, new taxon), 2 C . upsaliensis, 1 C . cryaerophila (Archobacter cryaerophilus, new taxon) from chicken ceca and 2 C . coli and 1 C . fennelliae (H . fennelliae) from raw chicken carcasses . Antimicrobial susceptibility testing of the different isolates against 9 antimicrobial agents was performed using the E-test . Overall, most isolates showed high to moderate susceptibility to gentamicin (97%), amoxicillin/clavulanate (95%), clindamycin (77%), chloramphenicol (77%), and ampicillin (69%) . Lower susceptibility were observed against tetracycline (49%), erythromycin (47%), ciprofloxacin (39%), and norfloxacin (36%) . This overall susceptibility profile generally applied to C . coli and C . jejuni, as well, although C . coli mostly showed higher susceptibility than C . jejuni . beta-lactamase production was detected in 59% of all the isolates, being higher in C . coli (72%) than C . jejuni (33%) . Whole cell protein profile analysis of 18 C . coli and 12 C . jejuni by SDS-PAGE revealed 6 different patterns . In both species, major variations existed in the region between mol wt 45-60 and all protein profiles were dominated by the presence of 5 major bands of mol wt: 61 (doublet), 45, 31 and an approximate 24 . Differences in banding patterns within and between both species indicated diversity and heterogeneity of strains . This study shows that despite high prevalence and diversity of strains in chicken, Campylobacter in Lebanon is rare in human diarrheic stools compared to Salmonella (3.2%) and Shigella (1.4%). Appl Environ Microbiol, 1999 Jun, 65(6), 2540 - 6 Generation of a superoxide dismutase (SOD)-deficient mutant of Campylobacter coli: evidence for the significance of SOD in Campylobacter survival and colonization; Purdy D et al.; The microaerophilic nature of Campylobacter species implies an inherent sensitivity towards oxygen and its reduction products, particularly the superoxide anion . The deleterious effects of exposure to superoxide radicals are counteracted by the activity of superoxide dismutase (SOD) . We have shown previously that Campylobacter coli possesses an iron cofactored SOD . The sodB gene of C . coli UA585 was insertionally inactivated by the site-specific insertion of a tetO cassette . Organisms harboring the inactivated gene failed to produce a biologically functional form of the enzyme . While the ability of this mutant to grow in aerobic conditions was unchanged relative to the parental strain, its survival was severely compromised when nongrowing cells were exposed to air . Accordingly, the SOD-deficient mutant was unable to survive for prolonged periods in model foods . Furthermore, inactivation of the sodB gene decreased the colonization potential in an experimental infection of 1-day-old chicks . In contrast, strain CK100, which is deficient in catalase activity, showed the same survival and colonization characteristics as the parental strain . These results indicate that SOD, but not catalase, is an important determinant in the ability of C . coli to survive aerobically and for optimal colonization within the chicken gut. Appl Environ Microbiol, 1999 Jun, 65(6), 2369 - 75 High-resolution genotyping of Campylobacter strains isolated from poultry and humans with amplified fragment length polymorphism fingerprinting; Duim B et al.; For epidemiological studies of Campylobacter infections, molecular typing methods that can differentiate campylobacters at the strain level are needed . In this study we used a recently developed genotyping method, amplified fragment length polymorphism (AFLP), which is based on selective amplification of restriction fragments of chromosomal DNA, for genetic typing of Campylobacter jejuni and Campylobacter coli strains derived from humans and poultry . We developed an automated AFLP fingerprinting method in which restriction endonucleases HindIII and HhaI were used in combination with one set of selective PCR primers . This method resulted in evenly distributed band patterns for amplified fragments ranging from 50 to 500 bp long . The discriminatory power of AFLP was assessed with a C . jejuni strain, an isogenic flagellin mutant, and distinct C . jejuni strains having known pulsed-field gel electrophoresis and fla PCR-restriction fragment length polymorphism genotypes . Unrelated C . jejuni strains produced heterogeneous patterns, whereas genetically related strains produced similar AFLP patterns . Twenty-five Campylobacter strains obtained from poultry farms in The Netherlands grouped in three C . jejuni clusters that were separate from a C . coli cluster . The band patterns of 10 C . jejuni strains isolated from humans were heterogeneous, and most of these strains grouped with poultry strains . Our results show that AFLP analysis can distinguish genetically unrelated strains from genetically related strains of Campylobacter species . However, desirable genetically related strains can be differentiated by using other genotyping methods . We concluded that automated AFLP analysis is an attractive tool which can be used as a primary method for subtyping large numbers of Campylobacter strains and is extremely useful for epidemiological investigations. J Food Prot, 1999 May, 62(5), 456 - 62 Development of a new medium for the isolation of Arcobacter spp; Johnson LG et al.; Arcobacter, the newly reclassified Campylobacter species, has been shown to cause diarrhea in both humans and animals . Few studies have been conducted regarding its occurrence in foods because of the lack of effective isolation and identification methods . The purpose of this study was to develop a plating medium that would be selective for the three most commonly found Arcobacter species . The effect of common components used in media intended for the isolation of Campylobacter, Helicobacter, and other gram-negative rods was examined . These components were divided into five distinct groups: (1) basic growth nutrients, (2) reducing and growth-promoting agents, (3) detoxifying agents, (4) antibiotics, and (5) color-enhancing compounds . Components from each of these groups were tested for their ability to recover Arcobacter on a solid medium when incubated aerobically at 30 degrees C for up to 72 h . Growth was evaluated by the ecometric technique, colony size, and differential colony morphology after incubation . After initial evaluations, five formulas showing the best results were selected and tested in detail and compared with brucella agar . A medium containing a basal nutrient mix along with 0.05% thioglycolic acid, 0.05% sodium pyruvate, and 5% sheep's blood (pH 6.9+/-0.2) was found to be the most effective for the growth of A . butzleri, A . cryaerophilus, and A . nitrofigilis . In addition to superior growth characteristics, a deep red color around the colonies also was observed with this formulation. J Neuroimmunol, 1999 May 3, 96(2), 245 - 50 Subclass distribution and the secretory component of serum IgA anti-ganglioside antibodies in Guillain-Barré syndrome after Campylobacter jejuni enteritis; Koga M et al.; Previously, we reported that IgA anti-GM1 antibody is more closely associated with preceding Campylobacter jejuni enteritis in Guillain-Barre syndrome (GBS) than are IgG and IgM antibodies . However, the mechanism of the induction of IgA anti-ganglioside antibodies is not clear . In this study, serum IgA antibodies against GM1, GM1b, and GD1a, and GalNAc-GD1a were examined in 152 GBS patients . In GBS, antecedent C . jejuni infection is closely associated with IgA antibodies, other than GM1, against GM1b . The IgA subclass distribution is completely restricted to IgA1, no secretory IgA anti-ganglioside antibody being detected . This result does not support the hypothesis that the serum IgA antibodies present in GBS after C . jejuni enteritis originate at mucosal sites, such as the gut mucosal immune system . Seventeen (85%) of 20 patients with IgA anti-ganglioside antibodies had serological evidence of C . jejuni infection and/or a history of antecedent diarrhea . Moreover, a motor nerve conduction study showed that patients with IgA antibodies frequently had axonal neuropathy, whereas none had demyelinating neuropathy . This may support the previous report that IgA isotype anti-GM1 antibodies are more closely associated with poor outcome than are the IgG or IgM isotypes . The induction mechanism of IgA anti-ganglioside antibodies must be clarified by determining whether concentrations of cytokines, which increase the IgA class switch, are elevated in patients with GBS after C . jejuni enteritis. Microbiol Immunol, 1999, 43(3), 241 - 4 Serum antibody level against GroEL type heat-shock protein of Campylobacter jejuni in patients with Guillain-Barré syndrome; Fujimoto S et al.; Recently there has been an increase in the number of cases reported of Guillain-Barre syndrome (GBS) developed after Campylobacter jejuni infection . To investigate the role of a C.jejuni GroEL-type heat-shock protein (CjHsp60) in the infection and induction of GBS, we examined the antibody level against CjHsp60 in 27 human sera, including GBS and non-GBS patients, by an enzyme-linked immunosorbent assay . Sera from patients with C . jejuni infection, despite the development of GBS, had a higher titer of anti-CjHsp60 antibody than those of patients without the infection and healthy control subjects . The patients with C . jejuni infection followed by GBS had slightly higher levels of this antibody than did the patients with infection who did not develop GBS, but there was no statistical significance . In conclusion, CjHsp60 is found to be one of the major immunogenic antigens in actual C . jejuni infection, but no evidence that supports the direct relationship between this protein and C . jejuni-associated GBS was found in this study. Zentralbl Veterinarmed B, 1999 Apr, 46(3), 181 - 8 Antibodies against Helicobacter felis in sera of cats and dogs; Seidel KE et al.; Serum samples from 61 dogs and 49 cats were screened for circulating antibodies against Helicobacter felis by an enzyme-linked immunosorbent assay (ELISA) using sonicated bacteria as an antigen . To improve the specificity of the ELISA, sera were absorbed with Campylobacter jejuni subsp . jejuni H . pylori as well as H . felis . Sera from 26 dogs (43%) and 19 cats (39%) revealed clear positive absorbance readings determined as an optical density (OD) that was statistically significant above the OD mean value {P < 0.025 (one-tailed); log10} . The absorbance pattern of ELISA-positive sera corresponded to results obtained with bovine and human reference sera . Furthermore, a correlation between the immune response and results from histopathological examination of gastric specimens from 22 dogs was demonstrated . It could be shown that antibodies against H . felis in sera of cats and dogs can easily be detected using an ELISA . The diagnostic value of this test must be evaluated in further investigations. Zentralbl Veterinarmed B, 1999 Apr, 46(3), 163 - 71 Biotin-streptavidin enzyme-linked immunosorbent assay for the detection of antibodies to Campylobacter jejuni and C . coli in chickens; Haas B et al.; An enzyme-linked immunosorbent assay (ELISA) was developed in a homologous system with bacterial ultrasonic-treated proteins as the antigen and antisera from chickens infected orally and subcutaneously with the strain Campylobacter jejuni serovar 6 (CJ 6) . The cut-off level was determined using antisera from non-infected specific-pathogen-free chickens up to the age of 10 weeks . The suitability of the ELISA system was verified using antisera taken from chickens orally infected at the age of 4 weeks with CJ 1, 6, 28 or 36 or with Campylobacter coli serovar 28 (CC 28) . The development of antibodies was monitored up to 6 weeks post-infection (p.i.) . Sera from chickens infected with CJ 1, 6, 36 or CC 28 contained specific antibodies to Campylobacter, whereas in those infected with CJ 28 no specific antibodies were found . Distinct cross-reactions were observed between CJ 6, 28 and CC 28 antigens and their antisera 6 weeks p.i., while poor cross-reactions were found with antisera to CJ 1 and 28 . Antibodies to strains of all heterologous serovars were successfully detected with an antigen pool comprised of CJ 1, 6 and 36 antigens . In 11 out of the 12 field sera obtained from 5- and 9-week-old broiler chickens suffering from campylobacteriosis, high specific antibody titres to Campylobacter jejuni were found. N Engl J Med, 1999 May 20, 340(20), 1525 - 32 Quinolone-resistant Campylobacter jejuni infections in Minnesota, 1992-1998 . Investigation Team; Smith KE et al.; BACKGROUND: Increasing resistance to quinolones among campylobacter isolates from humans has been reported in Europe and Asia, but not in the United States . We evaluated resistance to quinolones among campylobacter isolates from Minnesota residents during the period from 1992 through 1998 . METHODS: All 4953 campylobacter isolates from humans received by the Minnesota Department of Health were tested for resistance to nalidixic acid . Resistant isolates and selected sensitive isolates were tested for resistance to ciprofloxacin . We conducted a case-comparison study of patients with ciprofloxacin-resistant Campylobacter jejuni isolated during 1996 and 1997 . Domestic chicken was evaluated as a potential source of quinolone-resistant campylobacter . RESULTS: The proportion of quinolone-resistant C . jejuni isolates from humans increased from 1.3 percent in 1992 to 10.2 percent in 1998 (P<0.001) . During 1996 and 1997, infection with quinolone-resistant C . jejuni was associated with foreign travel and with the use of a quinolone before the collection of stool specimens . However, quinolone use could account for no more than 15 percent of the cases from 1996 through 1998 . The number of quinolone-resistant infections that were acquired domestically also increased during the period from 1996 through 1998 . Ciprofloxacin-resistant C . jejuni was isolated from 14 percent of 91 domestic chicken products obtained from retail markets in 1997 . Molecular subtyping showed an association between resistant C . jejuni strains from chicken products and domestically acquired infections in Minnesota residents . CONCLUSIONS: The increase in quinolone-resistant C . jejuni infections in Minnesota is largely due to infections acquired during foreign travel . However, the number of quinolone-resistant infections acquired domestically has also increased, largely because of the acquisition of resistant strains from poultry . The use of fluoroquinolones in poultry, which began in the United States in 1995, has created a reservoir of resistant C . jejuni. J Clin Microbiol, 1999 Jun, 37(6), 2084 - 6 Septic shock due to Helicobacter fennelliae in a non-human immunodeficiency virus-infected heterosexual patient; Hsueh PR et al.; Helicobacter fennelliae (formerly Campylobacter fennelliae) has been reported to cause bacteremia in homosexual men with or without human immunodeficiency virus (HIV) infection . We report here a 48-year-old, non-HIV-infected, heterosexual man with diabetes mellitus and cirrhosis of the liver who developed bacteremia and septic shock due to H . fennelliae . The patient was treated successfully initially with intravenous ampicillin-sulbactam and ceftazidime, followed by ampicillin-sulbactam only . These agents were active in vitro against the isolate by E-test results . To our knowledge, this is the first documented case of septic shock due to H . fennelliae in a non-HIV-infected, heterosexual, immunocompromised patient. J Clin Microbiol, 1999 Jun, 37(6), 1790 - 6 Rapid identification of thermotolerant Campylobacter jejuni, Campylobacter coli, Campylobacter lari, and Campylobacter upsaliensis from various geographic locations by a GTPase-based PCR-reverse hybridization assay; van Doorn LJ et al.; Recently, a gene from Campylobacter jejuni encoding a putative GTPase was identified . Based on two semiconserved GTP-binding sites encoded within this gene, PCR primers were selected that allow amplification of a 153-bp fragment from C . jejuni, C . coli, C . lari, and C . upsaliensis . Sequence analysis of these PCR products revealed consistent interspecies variation, which allowed the definition of species-specific probes for each of the four thermotolerant Campylobacter species . Multiple probes were used to develop a line probe assay (LiPA) that permits analysis of PCR products by a single reverse hybridization step . A total of 320 reference strains and clinical isolates from various geographic origins were tested by the GTP-based PCR-LiPA . The PCR-LiPA is highly specific in comparison with conventional identification methods, including biochemical and whole-cell protein analyses . In conclusion, a simple method has been developed for rapid and highly specific identification of thermotolerant Campylobacter species. J Bacteriol, 1999 May, 181(10), 3298 - 302 A novel Campylobacter jejuni two-component regulatory system important for temperature-dependent growth and colonization; Br inverted question markas AM et al.; Campylobacter jejuni colonizes the intestines of domestic and wild animals and is a common cause of human diarrheal disease . We identified a two-component regulatory system, designated the RacR-RacS (reduced ability to colonize) system, that is involved in a temperature-dependent signalling pathway . A mutation of the response regulator gene racR reduced the organism's ability to colonize the chicken intestinal tract and resulted in temperature-dependent changes in its protein profile and growth characteristics. Poult Sci, 1999 Apr, 78(4), 536 - 45 Natural and experimental infections of Arcobacter in poultry; Wesley IV et al.; Arcobacter butzleri causes human enteritis and is frequently recovered from poultry carcasses . The purpose of this study was to determine 1) the natural distribution of A . butzleri in poultry and 2) its relative pathogenicity in experimentally infected poultry . Cloacal samples (n = 407) were collected on four occasions from three flocks of chickens . Overall, Arcobacter spp . were recovered from 15% of the birds; A . butzleri was identified in 1% of cloacal samples . Three experimental trials were conducted to determine the susceptibility of birds . In Trial 1, 3-d-old chicks (n = 62) were divided into three groups and infected per os with 1) A . butzleri American Type Culture Collection (ATCC) 49616, 2) a suspension of four field strains of A . butzleri isolated from retail purchased chicken, and 3) Campylobacter jejuni (positive control) . Arcobacter was not detected in cloacal swabs or in cecal samples of chicks through Day 5 postinfection; C . jejuni was detected in cloacal swabs of all positive control birds . In Trial 2, 5-d-old outbred turkey poults (n = 88) were infected as described above with the addition of a group infected with a suspension of four field strains of A . butzleri from turkey meat . Arcobacter butzleri was recovered from either cloacal swabs or cecal contents of only 6.0% of birds (4 of 67); C . jejuni was recovered from 100% of the positive control birds (n = 21) . In Trial 3, 3-d-old turkey poults of the highly inbred Beltsville White strain (n = 141) were experimentally inoculated . In contrast to earlier trials, A . butzleri was recovered overall from the cloacal swabs or tissues of 65% of the turkeys. Res Microbiol, 1999 Apr, 150(3), 213 - 9 Study of the presence of Campylobacter jejuni and C . coli in sand samples from four Swiss chicken farms; Studer E et al.; Chicken farms are frequently infected with Campylobacter jejuni and Campylobacter coli . The objective of the present study was to investigate environmental samples from chicken farms for the presence of C . jejuni and C . coli . Every week between July and November 1997, three sand samples from the runs of four chicken farms were analyzed by culture and directly by polymerase chain reaction (PCR) . These two detection methods were compared to each other . A total of 231 samples were tested . Eleven samples (4%) were found to contain Campylobacter cells by culture, whereas 157 samples (68%) were positive by PCR . All samples which were positive by culture were also positive by PCR . All direct PCR products were further typed by restriction fragment length polymorphism (RFLP) . Three different RFLP types and mixtures of these types were observed . Direct PCR products of one chicken farm were further typed by direct sequencing and two temporally separated sequence types could be distinguished . Campylobacter strains isolated by culture were also typed by RFLP and direct sequencing revealing close accordance with the corresponding direct PCR products. J Med Microbiol, 1999 May, 48(5), 461 - 9 Different invasion phenotypes of Campylobacter isolates in Caco-2 cell monolayers; Harvey P et al.; The pathogenesis of campylobacter enteritis is not well understood, but invasion into and translocation across intestinal epithelial cells may be involved in the disease process, as demonstrated for a number of other enteric pathogens . However, the mechanisms involved in these processes are not clearly defined for campylobacters . In this study, isolates were compared quantitatively in established assays with the enterocyte-like cell line, Caco-2, to determine the extent to which intracellular invasion contributes to translocation across epithelial cell monolayers, and whether isolates vary in this respect . Ten fresh Campylobacter isolates were compared and shown to differ in invasiveness by a factor of 10-fold by following their recovery from gentamicin-treated Caco-2 cells grown on nonpermeable tissue-culture wells . Four of these isolates with contrasting invasive ability were also shown to vary in their ability to translocate across Caco-2 cells grown on semipermeable Transwell inserts by a factor >10 . However, translocation did not quantitatively correlate with the intracellular invasiveness of these isolates . Isolate no . 9752 was poorly invasive but had modest translocation ability, isolate no . 10392 was very invasive but did not translocate significantly and remained within the monolayer, isolate no . 9519 both translocated and invaded well, whereas, isolate no . 235 translocated very efficiently but was poorly invasive . Isolate no . 9519 also uniquely caused a transitory flattening of the Caco-2 cells and a possible drop in trans-epithelial electrical resistance (TEER) of the Transwell monolayers, whereas isolate no . 235 did not show these effects . Together these data demonstrate that there are significantly different 'invasion' phenotypes among Campylobacter strains involving different degrees of intracellular invasion, and either different rates of transcellular trafficking or, alternatively, paracellular trafficking. Infect Immun, 1999 May, 67(5), 2433 - 40 Identification of virulence genes of Helicobacter pylori by random insertion mutagenesis; Bijlsma JJ et al.; The complete genome of the gram-negative bacterial pathogen Helicobacter pylori, an important etiological agent of gastroduodenal disease in humans, has recently been published . This sequence revealed that the putative products of roughly one-third of the open reading frames (ORFs) have no significant homology to any known proteins . To be able to analyze the functions of all ORFs, we constructed an integration plasmid for H . pylori and used it to generate a random mutant library in this organism . This integration plasmid, designated pBCalpha3, integrated randomly into the chromosome of H . pylori . To test the capacity of this library to identify virulence genes, subsets of this library were screened for urease-negative mutants and for nonmotile mutants . Three urease-negative mutants in a subset of 1,251 mutants (0.25%) and 5 nonmotile mutants in a subset of 180 mutants (2.7%) were identified . Analysis of the disrupted ORFs in the urease-negative mutants revealed that two had disruptions of genes of the urease locus, ureB and ureI, and the third had a disruption of a unrelated gene; a homologue of deaD, which encodes an RNA helicase . Analysis of the disrupted ORFs in the nonmotile mutants revealed one ORF encoding a homologue of the paralyzed flagellar protein, previously shown to be involved in motility in Campylobacter jejuni . The other four ORFs have not been implicated in motility before . Based on these data, we concluded that we have generated a random insertion library in H . pylori that allows for the functional identification of genes in H . pylori. Appl Environ Microbiol, 1999 May, 65(5), 2272 - 5 Stability of related human and chicken Campylobacter jejuni genotypes after passage through chick intestine studied by pulsed-field gel electrophoresis; Hanninen ML et al.; The genomic stability of 12 Campylobacter jejuni strains consisting of two groups of human and chicken isolates was studied by analysis of their PFGE (pulsed-field gel electrophoresis) patterns after passage through newly hatched chicks' intestines . The patterns of SmaI, SalI, and SacII digests remained stable after intestinal passage, except for those of two strains . One originally human strain, FB 6371, changed its genotype from II/A (SmaI/SacII) to I/B . Another strain, BTI, originally isolated from a chicken, changed its genotype from I/B to a new genotype . The genomic instability of the strains was further confirmed by SalI digestion and ribotyping of the HaeIII digests . In addition, heat-stable serotype 57 of strain FB 6371 changed to serotype 27 in all isolates with new genotypes but remained unchanged in an isolate with the original genotype . Serotype 27 of strain BTI remained stable . Our study suggests that during intestinal colonization, genomic rearrangement, as demonstrated by changed PFGE and ribopatterns, may occur. J Neurol Sci, 1999 Feb 1, 163(1), 53 - 7 Are Campylobacter curvus and Campylobacter upsaliensis antecedent infectious agents in Guillain-Barré and Fisher's syndromes? Koga M, Yuki N, Takahashi M, Saito K, Hirata K. Campylobacter curvus and Campylobacter upsaliensis were isolated from stools of patients with Guillain-Barre (GBS) or Fisher's (FS) syndromes . Whether these microorganisms are pathogens of antecedent diarrhea in GBS and FS is not clear, therefore, we made a serological examination . There were no differences in antibody titer to these organisms among the patients with GBS, FS, and the controls . Some patients had elevated antibodies to the bacteria, but most also had serological evidence of C . jejuni infection . Moreover, the patients from whom C . curvus had been isolated did not have antibodies to the bacterium, indicative that they were healthy carriers of C . curvus or that the isolates were the product of contamination . We conclude that neither C . curvus nor C . upsaliensis is the major agent of antecedent diarrhea in GBS and FS. FEMS Microbiol Lett, 1999 Apr 1, 173(1), 77 - 84 High-resolution genomic fingerprinting of Campylobacter jejuni and Campylobacter coli by analysis of amplified fragment length polymorphisms; Kokotovic B et al.; A method for high-resolution genomic fingerprinting of the enteric pathogens Campylobacter jejuni and Campylobacter coli, based on the determination of amplified fragment length polymorphism, is described . The potential of this method for molecular epidemiological studies of these species is evaluated with 50 type, reference, and well-characterised field strains . Amplified fragment length polymorphism fingerprints comprised over 60 bands detected in the size range 35-500 bp . Groups of outbreak strains, replicate subcultures, and 'genetically identical' strains from humans, poultry and cattle, proved indistinguishable by amplified fragment length polymorphism fingerprinting, but were differentiated from unrelated isolates . Previously unknown relationships between three hippurate-negative C . jejuni strains, and two C . coli var . hyoilei strains, were identified . These relationships corresponded to available epidemiological data . We conclude that this amplified fragment length polymorphism fingerprinting method may be a highly effective tool for molecular epidemiological studies of Campylobacter spp. Mol Microbiol, 1999 Apr, 32(1), 131 - 8 Helicobacter pylori with separate beta- and beta'-subunits of RNA polymerase is viable and can colonize conventional mice; Raudonikiene A et al.; The genes encoding the beta- and beta'-subunits of RNA polymerase (rpoB and rpoC respectively) are fused as one continuous open reading frame in Helicobacter pylori and in other members of this genus, but are separate in other bacterial taxonomic groups, including the closely related genus Campylobacter . To test whether this beta-beta' tethering is essential, we used polymerase chain reaction-based cloning to separate the rpoB and rpoC moieties of the H . pylori rpoB-rpoC fusion gene with a non-polar chloramphenicol resistance cassette containing a new translational start, and introduced this construct into H . pylori by electro-transformation . H . pylori containing these separated rpoB and rpoC genes in place of the native fusion gene produced non-tethered beta and beta' RNAP subunits, grew well in culture and colonized and proliferated well in conventional C57BL/6 mice . Thus, the extraordinary beta-beta' tethering is not essential for H . pylori viability and gastric colonization. Zentralbl Veterinarmed B, 1999 Mar, 46(2), 141 - 4 Survival times of Campylobacter coli in sterilized buffalo milk; Tresierra-Ayala A et al.; The survival level of five Campylobacter coli strains isolated from bovine faeces was studied in sterilized buffalo milk kept at 4 degrees C under aerobic conditions . All strains died between 21 and 30 h of storage. Lett Appl Microbiol, 1999 Apr, 28(4), 285 - 90 The use of hipO, encoding benzoylglycine amidohydrolase (hippuricase), as a reporter of gene expression in Campylobacter coli; Park SF; A novel integrative promoter probe vector which utilizes the Campylobacter jejuni hipO gene as a reporter of gene expression was developed as a genetic tool in Campylobacter coli . The utility of the system was demonstrated by coupling expression of the hipO reporter to the promoters of flaA, flaB and katA . Subsequently, expression of these genes could be monitored accurately from chromosomally-borne transcriptional fusions using a simple non-destructive colorimetric assay . The system should serve as a useful tool for studying the response of Camp . coli to environmental stresses. MMWR Morb Mortal Wkly Rep, 1999 Mar 12, 48(9), 189 - 94 Incidence of foodborne illnesses: preliminary data from the Foodborne Diseases Active Surveillance Network (FoodNet)--United States, 1998; Differentiation of Campylobacter coli and C . jejuni by length and DNA sequence of the 16S-23S rRNA internal spacer region; Department of Veterinary Microbiology and The Danish Centre for Advanced Food Studies, Royal Veterinary and Agricultural University, 1870 Frederiksberg C . kvlhc@unidph.uni-c.dk The internal spacer region (ISR) between the 16S and 23S rRNA genes of Campylobacter was investigated by PCR fragment length typing and DNA sequencing of clinical and chicken wild-type isolates . PCR fragment length typing showed one fragment of 859 nt in length for the 12 strains of Campylobacter coli investigated . Thirty-six of the Campylobacter jejuni subsp . jejuni strains possessed one fragment, which varied in size between 727 and 802 nt . Three strains showed two fragments between 501 and 923 nt . Strains of C . jejuni subsp . doylei, Campylobacter lari and Campylobacter upsaliensis possessed one or two fragments with lengths different from those of C . coli and C . jejuni subsp . jejuni . DNA sequences were obtained from 54 nt downstream of rrs up to rrl of four strains of C . coli, eight strains of C . jejuni subsp . jejuni, and one strain each of C . jejuni subsp . doylei and C . lari, selected to represent the different biotypes of Campylobacter . ISR lengths determined by PCR fragment length typing and DNA sequencing corresponded for 12 strains . For two strains of C . coli, PCR fragment length typing underestimated ISR lengths by 159 and 193 nt, probably related to incomplete resolution of the distal helical structures, which were not fully denatured during PAGE . For the 14 strains and the published C . jejuni subsp . jejuni sequence, the first 206-211 nt were conserved and included the two tRNA genes in the characteristic tRNA(Ala) to tRNA(Ile) order separated by a short 8-9 nt spacer region . Within the region downstream of tRNA(Ile) conserved regions were identified which allowed a separation of C . lari from C . coli and C . jejuni but not separation of C . coli from C . jejuni . The 69-282 nt longer variable regions in C . coli strains allowed separation of this species from C . jejuni, confirming results obtained by PCR typing . Certain nucleic acid positions in variable regions were related to the Lior biotypes . Sequence information from ISRs of more strains is needed to ascertain if separation of species and biotypes will be possible for diagnostic purposes. Microbiology, 1999 Jan, 145 ( Pt 1), 89 - 98 Cloning, sequencing and molecular analysis of the Campylobacter jejuni groESL bicistronic operon; Thies FL et al.; The groESL bicistronic operon from the enteric pathogen Campylobacter jejuni was cloned and sequenced . It consists of two ORFs encoding proteins with molecular masses of 9.5 and 57.9 kDa, which showed a high degree of homology to other bacterial GroES and GroEL proteins . Northern blot analysis suggested that the groESL operon is transcribed as a bicistronic mRNA, and its steady-state level was markedly increased after temperature upshift . By primer extension assay, one potential transcription start point preceding the groESL genes could be demonstrated, and a putative promoter region compatible with both Escherichia coli and C . jejuni sigma70 consensus sequences was identified . A conserved inverted repeat, which is believed to be involved in the regulation of the groESL genes, was found between the -10 promoter box and the groES translation start site . The complete coding region of groEL was fused with pET-22b(+) and expressed in E . coli as a His6-tagged recombinant protein (rCjHsp60-His) . After purification, the protein was recognized by an anti-HSP60 monoclonal antibody . ELISA and Western immunoblotting experiments showed that IgG and IgA antibody responses against rCjHsp60-His were not significantly increased in sera from 24 patients with sporadic Campylobacter infection when compared to sera from 16 healthy controls. BMJ, 1999 Apr 17, 318(7190), 1046 - 50 Study of infectious intestinal disease in England: rates in the community, presenting to general practice, and reported to national surveillance . The Infectious Intestinal Disease Study Executive; Wheeler JG et al.; OBJECTIVE: To establish the incidence and aetiology of infectious intestinal disease in the community and presenting to general practitioners . Comparison with incidence and aetiology of cases reaching national laboratory based surveillance . DESIGN: Population based community cohort incidence study, general practice based incidence studies, and case linkage to national laboratory surveillance . SETTING: 70 general practices throughout England . PARTICIPANTS: 459 975 patients served by the practices . Community surveillance of 9776 randomly selected patients . MAIN OUTCOME MEASURES: Incidence of infectious intestinal disease in community and reported to general practice . RESULTS: 781 cases were identified in the community cohort, giving an incidence of 19.4/100 person years (95% confidence interval 18.1 to 20.8) . 8770 cases presented to general practice (3.3/100 person years (2.94 to 3.75)) . One case was reported to national surveillance for every 1.4 laboratory identifications, 6.2 stools sent for laboratory investigation, 23 cases presenting to general practice, and 136 community cases . The ratio of cases in the community to cases reaching national surveillance was lower for bacterial pathogens (salmonella 3.2:1, campylobacter 7.6:1) than for viruses (rotavirus 35:1, small round structured viruses 1562:1) . There were many cases for which no organism was identified . CONCLUSIONS: Infectious intestinal disease occurs in 1 in 5 people each year, of whom 1 in 6 presents to a general practitioner . The proportion of cases not recorded by national laboratory surveillance is large and varies widely by microorganism . Ways of supplementing the national laboratory surveillance system for infectious intestinal diseases should be considered. J Clin Microbiol, 1999 May, 37(5), 1646 - 50 Cytolethal distending toxin genes in Campylobacter jejuni and Campylobacter coli isolates: detection and analysis by PCR; Eyigor A et al.; Campylobacter jejuni produces a toxin called cytolethal distending toxin (CDT) . Knowledge of the prevalence and homogeneity of Campylobacter sp . cdt genes is incomplete . In this work, we identified four PCR primer pairs that collectively amplified cdt genes in all of the C . jejuni and Campylobacter coli strains tested . Restriction analyses of the cdt PCR products showed clear differences between the cdt genes of these two species, yet there were few heterogeneities noted between members of the same species . Consequently, it may be possible to speciate C . jejuni and C . coli isolates on the basis of restriction patterns within their cdt genes. J Clin Microbiol, 1999 May, 37(5), 1319 - 23 Recurrent "Flexispira rappini" bacteremia in an adult patient undergoing hemodialysis: case report; Sorlin P et al.; A blood culture from a 65-year-old febrile man undergoing hemodialysis revealed, 5 days after inoculation, an unusual gram-negative fusiform rod with darting motility . During another episode of fever 21 days later, this Campylobacter-like organism was again recovered from three blood cultures and subcultured under an H2-enriched microaerobic atmosphere . The organism was catalase negative and oxidase positive and hydrolyzed urea rapidly . Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of whole-cell proteins was indistinguishable from that of "Flexispira rappini" LMG 8738 described by Archer et al . in 1988 (J . R . Archer, S . Romero, A . E . Ritchier, M . E . Hamacher, B . M . Steiner, J . H . Bryner, and R . F . Schell, J . Clin . Microbiol . 26:101-105, 1988) . The analysis of the 16S ribosomal DNA sequence revealed a similarity of 99.3% between the two strains . The patient recovered completely after a 4-week course of meropenem therapy . This is the first reported case of a recurrent "F . rappini" bacteremia in an adult patient, which confirms that this organism may be an invasive pathogen in immunocompromised patients, like other newly described Helicobacter species. J Immunol, 1999 Apr 15, 162(8), 4773 - 80 Actinobacillus actinomycetemcomitans immunosuppressive protein is a member of the family of cytolethal distending toxins capable of causing a G2 arrest in human T cells; Shenker BJ et al.; We have previously shown that Actinobacillus actinomycetecomitans produces an immunosuppressive factor (ISF) capable of impairing human lymphocyte function by perturbing cell cycle progression . We now report that ISF is the product of the cdtB gene, one of three genes encoding the family of cytolethal distending toxins (Cdt) . The ISF polypeptide exhibits >/=95% identity with Hemophilus ducreyi CdtB protein and </=60% homology with Escherichia coli or Campylobacter jejuni CdtB . Pretreatment of PHA-activated lymphocytes with 5-25 ng ISF results in G2 arrest of CD4+ and CD8+ T cells . Similarly, treatment of HeLa cells results in G2 arrest and cell elongation and distension . However, lymphocytes are at least 5 times more sensitive to ISF than HeLa cells and do not undergo the elongation and distension that characterizes interactions of Cdts with cell lines . ISF-treated lymphocytes express normal cyclin A and B1 levels, but contain reduced levels of cell cycle-dependent kinase-1 (Cdk1) . Additionally, the majority of Cdk1 is in the hyperphosphorylated, inactive, form . In contrast, PHA-induced G2 cells contain elevated levels of the hypophosphorylated, active Cdk1 . Failure of ISF-treated cells to dephosphorylate Cdk1 is not associated with decreased availability of Cdc25 . These studies suggest that the CdtB protein alone is capable of inducing G2 arrest in lymphocytes and cell cycle arrest, elongation, and distension of HeLa cells . Our studies also suggest that lymphocytes may be primary targets for A . actinomycetemcomitans CdtB (ISF) and possibly for other Cdt family members as well . Thus, Cdts may function to impair host immunity and contribute to the pathogenesis of disease associated with Cdt-producing organisms. J Bacteriol, 1999 Apr, 181(8), 2501 - 6 Cloning and characterization of two bistructural S-layer-RTX proteins from Campylobacter rectus; Braun M et al.; Campylobacter rectus is an important periodontal pathogen in humans . A surface-layer (S-layer) protein and a cytotoxic activity have been characterized and are thought to be its major virulence factors . The cytotoxic activity was suggested to be due to a pore-forming protein toxin belonging to the RTX (repeats in the structural toxins) family . In the present work, two closely related genes, csxA and csxB (for C . rectus S-layer and RTX protein) were cloned from C . rectus and characterized . The Csx proteins appear to be bifunctional and possess two structurally different domains . The N-terminal part shows similarity with S-layer protein, especially SapA and SapB of C . fetus and Crs of C . rectus . The C-terminal part comprising most of CsxA and CsxB is a domain with 48 and 59 glycine-rich canonical nonapeptide repeats, respectively, arranged in three blocks . Purified recombinant Csx peptides bind Ca2+ . These are characteristic traits of RTX toxin proteins . The S-layer and RTX domains of Csx are separated by a proline-rich stretch of 48 amino acids . All C . rectus isolates studied contained copies of either the csxA or csxB gene or both; csx genes were absent from all other Campylobacter and Helicobacter species examined . Serum of a patient with acute gingivitis showed a strong reaction to recombinant Csx protein on immunoblots. Lett Appl Microbiol, 1999 Mar, 28(3), 233 - 7 Identification of a gene encoding an immuno-reactive membrane protein from Campylobacter jejuni; Connerton PL et al.; A gene encoding a putative membrane protein has been identified from Campylobacter jejuni NCTC 11168 following an immuno-screen of a lambda ZAP II genomic DNA library with antiserum raised against glycine-extractable proteins . The nucleotide sequence of the entire genomic insert revealed six open reading frames, all but one of which have sequence homologues in the complete genome sequence of Helicobacter pylori . The gene encoding the immuno-reactive protein was further identified by independent expression of these reading frames in Escherichia coli . The gene encodes an integral membrane protein, expression of which in E . coli results in a profound filamentous phenotype. Lett Appl Microbiol, 1999 Mar, 28(3), 194 - 8 Morphological changes of synchronized Campylobacter jejuni populations during growth in single phase liquid culture; Thomas C et al.; Campylobacter jejuni strains demonstrate a variety of growth phase-linked distinct morphological forms when grown in liquid culture . The typical spiral form of the organism, evident during logarithmic phase, undergoes elongation during stationary phase before becoming coccoid via the formation of membrane blebs and budded forms in decline phase . Cellular elongation and coccoid formation occurred despite the inhibition of protein synthesis and without a detectable change in the protein components of the inner and outer cell membranes. Gene, 1999 Apr 1, 230(1), 61 - 7 The ClpB protein from Campylobacter jejuni: molecular characterization of the encoding gene and antigenicity of the recombinant protein; Thies FL et al.; The ClpB heat-shock protein is necessary for the survival of Escherichia coli cells upon sudden increase of temperature . Using a PCR-based genomic walking method, the nucleotide sequence of a clpB homolog from Campylobacter jejuni was determined . The clpB gene encodes a protein of 857 amino acid (aa) residues, with a predicted molecular mass of 95.3kDa . Alignment of the deduced aa sequence with other known bacterial ClpB proteins revealed overall identity from 47% (E . coli) to 61% (Helicobacter pylori) . Within the clpB promoter region, as indicated by primer extension analysis, we identified a sequence identical to the E . coli sigma70 consensus promoter . Northern blot analysis confirmed that clpB is heat-inducible in C . jejuni . The ClpB protein, fused to a 6xHis tag, was synthesized in E . coli and purified by metal-affinity and size exclusion chromatography . In ELISA studies, IgA levels reactive to recombinant ClpB were significantly higher in sera of patients with prior C . jejuni infections than in sera obtained from healthy control persons. J Infect Dis, 1999 May, 179(5), 1183 - 9 Ganglioside GM1 mimicry in Campylobacter strains from sporadic infections in the United States; Nachamkin I et al.; To determine whether GM1-like epitopes in Campylobacter species are specific to O serotypes associated with Guillain-Barre syndrome (GBS) or whether they are frequent among random Campylobacter isolates causing enteritis, 275 random enteritis-associated isolates of Campylobacter jejuni were analyzed . To determine whether GM1-like epitopes in Campylobacter species are specific to O serotypes associated with Guillan-Barre syndrome (GBS) or whether they are frequent among random Campylobacter isolates causing enteritis, 275 enteritis-associated isolates, randomly collected in the United States, were analyzed using a cholera-toxin binding assay {corrected} . Overall, 26.2% of the isolates were positive for the GM1-like epitope . Of the 36 different O serotypes in the sample, 21 (58.3%) contained no strains positive for GM1, whereas in 6 serotypes (16.7%), >50% of isolates were positive for GM1 . GBS-associated serotypes were more likely to contain strains positive for GM1 than were non-GBS-associated serotypes (37.8% vs . 15.1%, P=.0116) . The results suggest that humans are frequently exposed to strains exhibiting GM1-like mimicry and, while certain serotypes may be more likely to possess GM1-like epitopes, the presence of GM1-like epitopes on Campylobacter strains does not itself trigger GBS. Appl Environ Microbiol, 1999 Apr, 65(4), 1636 - 43 Detection of small numbers of Campylobacter jejuni and Campylobacter coli cells in environmental water, sewage, and food samples by a seminested PCR assay; Waage AS et al.; A rapid and sensitive assay was developed for detection of small numbers of Campylobacter jejuni and Campylobacter coli cells in environmental water, sewage, and food samples . Water and sewage samples were filtered, and the filters were enriched overnight in a nonselective medium . The enrichment cultures were prepared for PCR by a rapid and simple procedure consisting of centrifugation, proteinase K treatment, and boiling . A seminested PCR based on specific amplification of the intergenic sequence between the two Campylobacter flagellin genes, flaA and flaB, was performed, and the PCR products were visualized by agarose gel electrophoresis . The assay allowed us to detect 3 to 15 CFU of C . jejuni per 100 ml in water samples containing a background flora consisting of up to 8, 700 heterotrophic organisms per ml and 10,000 CFU of coliform bacteria per 100 ml . Dilution of the enriched cultures 1:10 with sterile broth prior to the PCR was sometimes necessary to obtain positive results . The assay was also conducted with food samples analyzed with or without overnight enrichment . As few as </=3 CFU per g of food could be detected with samples subjected to overnight enrichment, while variable results were obtained for samples analyzed without prior enrichment . This rapid and sensitive nested PCR assay provides a useful tool for specific detection of C . jejuni or C . coli in drinking water, as well as environmental water, sewage, and food samples containing high levels of background organisms. Appl Environ Microbiol, 1999 Apr, 65(4), 1501 - 5 Detection of cytolethal distending toxin activity and cdt genes in Campylobacter spp . isolated from chicken carcasses; Eyigor A et al.; This study was designed to determine whether isolates from chicken carcasses, the primary source of Campylobacter jejuni and Campylobacter coli in human infections, commonly carry the cdt genes and also whether active cytolethal distending toxin (CDT) is produced by these isolates . Campylobacter spp . were isolated from all 91 fresh chicken carcasses purchased from local supermarkets . Campylobacter spp . were identified on the basis of both biochemical and PCR tests . Of the 105 isolates, 70 (67%) were identified as C . jejuni, and 35 (33%) were identified as C . coli . PCR tests amplified portions of the cdt genes from all 105 isolates . Restriction analysis of PCR products indicated that there appeared to be species-specific differences between the C . jejuni and C . coli cdt genes, but that the restriction patterns of the cdt genes within strains of the same species were almost invariant . Quantitation of active CDT levels produced by the isolates indicated that all C . jejuni strains except four (94%) had mean CDT titers greater than 100 . Only one C . jejuni strain appeared to produce no active CDT . C . coli isolates produced little or no toxin . These results confirm the high rate of Campylobacter sp . contamination of fresh chicken carcasses and indicate that cdt genes may be universally present in C . jejuni and C . coli isolates from chicken carcasses. J Periodontol, 1999 Feb, 70(2), 131 - 8 Microbiota of successful osseointegrated dental implants; Lee KH et al.; BACKGROUND: The long-term survival of dental implants depends, in part, on control of bacterial infection in the peri-implant region . Periodontal pathogens colonized implants symptomatic through infection, whereas the microbiota of successful implants was similar to that of periodontal health . This study examined the impact on the peri-implant microbiota of crown restorations; implant type; length of time of loading; history of implant or periodontal infections; and whether implants replaced single or multiple teeth . It was of particular interest to evaluate implant colonization by species in a newly described red complex of periodontal pathogens, Porphyromonas gingivalis and Bacteroides forsythus . METHODS: This study sampled 43 partially edentulous subjects with successfully osseointegrated titanium root-form dental implants . Eighty-one (81) non-submerged and 20 submerged asymptomatic implants, 83 crowned, and 36 uncrowned teeth were sampled from peri-implant or subgingival sites . The microbiota of samples was evaluated using whole genomic DNA probes in a checkerboard assay to 23 subgingival species . RESULTS: Implants were colonized principally by oral streptococci, capnocytophagae, Veillonella parvula, Peptostreptococcus micros, and Fusobacterium nucleatum . The periodontal species, P . gingivalis, B . forsythus, Prevotella intermedia, Prevotella nigrescens, and Campylobacter rectus were detected in a few subjects . The microbiota around crowned implants and crowned teeth was similar . Streptococcus oralis, P . intermedia, and Selenomonas noxia were elevated in samples from uncrowned teeth compared to crowned teeth and implants . Microbial complexity increased as loading time increased, but colonization by periodontal pathogens, including red complex species, was higher in subjects with previous periodontal disease . No differences were observed in the microbiota of 1- and 2-stage implants, or between implants supporting single or multiple restorations . CONCLUSIONS: While presence of crowns had only a minor impact on the peri-implant microbiota, microbial changes were observed the longer the implants had been in function and in those patients with a history of periodontal or peri-implant infections . A history of periodontitis had a greater impact on the peri-implant microbiota than implant loading time . The major influence on the peri-implant microbiota was, however, the microbiota on remaining teeth . P . gingivalis and B . forsythus, red complex periodontal pathogens, colonized several implants, although all implants were successfully osseointegrated. Spine, 1999 Mar 15, 24(6), 582 - 4 Pyogenic vertebral osteomyelitis caused by Campylobacter fetus subspecies fetus . A case report; Yamashita K et al.; STUDY DESIGN: Clinical observation of a patient . OBJECTIVES: To present the clinical features of an unusual infection of the spine caused by Campylobacter fetus subspecies fetus and to suggest treatment . SUMMARY OF BACKGROUND DATA: This is only the second reported case of pyogenic vertebral osteomyelitis caused by Campylobacter fetus subspecies fetus . METHODS: A 66-year-old man had pain of the left lower extremity . Radiologic examination revealed an epidural mass associated with destruction of the L5-S1 vertebral bodies . RESULTS: Biopsy of the epidural mass was performed, and culture yielded Campylobacter fetus subspecies fetus . After intravenous antibiotics, oral doxycycline and erythromycin were given for 5 months . At 9 months after antibiotic treatment was completed, the patient's condition was stable . CONCLUSIONS: Prolonged oral administration of doxycycline and erythromycin was curative in this patient. Cent Afr J Med, 1998 Sep, 44(9), 223 - 9 Distributional patterns of bacterial diarrhoeagenic agents and antibiograms of isolates from diarrhoeaic and non-diarrhoeaic patients in urban and rural areas of Nigeria; Obi CL et al.; OBJECTIVES: To determine the prevalence of bacteria that could cause diarrhoea in stool specimens of individuals with and without diarrhoea in both urban and rural areas of Nigeria . To ascertain the antibiotic susceptibilities of the bacterial diarrhoeagenic agents isolated . To document the predominant signs and symptoms associated with the various bacterial agents of diarrhoea . DESIGN: Prospective study . SETTING: Patients/individuals attending government and private clinics in Lagos, Edo and Cross-River States of Nigeria . SUBJECTS: A total of 1,200 stool samples were collected from patients with diarrhoea . Another total of 1,200 stool specimens were obtained from controls . RESULTS: For diarrhoea cases in urban areas Campylobacter spp . were more predominant (28%) and were followed by enteropathogenic Escherichia coli (EPEC) (28%) whereas in rural areas, EPEC were the most commonly isolated bacteria (18%), closely followed by Salmonella spp . (16%) . Controls had a similar distribution pattern . Higher rates of isolation of these enteric bacteria were recorded among diarrhoea cases than in controls (p < 0.05) . Diarrhoea due to Vibrio, Yersinia, Aeromonas, Plesiomonas and EPEC was mainly watery whereas it mainly consisted of blood/mucus for Shigella and Salmonella . All were associated with abdominal pain and fever . Results presented also indicate that over 80% of Shigella species, Salmonella, EPEC and P . shigelloides were susceptible to nalidixic acid and nitrofurantoin . Virtually all the enteropathogens were resistant to commonly used antibiotics such ampicillin, erythromycin, tetracyclines and streptomycin . CONCLUSION: Results show that distributional patterns of bacterial agents of diarrhoea may vary in urban and rural areas and have revealed the effectiveness of nalidixic acid, gentamicin and nitrofurantoin, in that order, against these enteropathogens. Epidemiol Infect, 1999 Feb, 122(1), 175 - 82 Clonality of Campylobacter sputorum bv . paraureolyticus determined by macrorestriction profiling and biotyping, and evidence for long-term persistent infection in cattle; On SL et al.; Eighteen strains of Campylobacter sputorum bv . paraureolyticus (isolated over a 12-month period from seven dairy cows contained in a single herd) were examined by resistotyping, and macrorestriction profiling using pulsed field gel electrophoresis (PFGE) . The resistotypes of these strains were identical, although repeat testing indicated resistance to metronidazole was not a reliable trait for typing purposes . Five SmaI-derived genotypes were identified among the 18 strains . In 5 of 7 cows, isolates obtained from the same animal, but from different time periods, were genotypically indistinguishable, indicating persistence of infection . Macrorestriction profiles of 5 strains representing the 5 SmaI genotypes and 8 other strains of C . sputorum from various sources, were prepared using 4 endonucleases (SmaI, SalI, BamHI and KpnI) . The only other strain of C . sputorum bv . paraureolyticus examined (a Canadian isolate from human faeces), was found to have a SmaI macrorestriction profile identical with one of the five clones isolated from the cattle . Moreover, SalI and BamHI profiles of all bv . paraureolyticus strains were similar, while digestion with KpnI was not observed . By contrast, the seven strains of C . sputorum bv . sputorum yielded various macrorestriction profiles with all the enzymes used, and features distinguishing the two biovars studied could be identified . This study indicates that C . sputorum can persist in cattle for at least 12 months and exhibits a clonal population genetic structure. Epidemiol Infect, 1999 Feb, 122(1), 15 - 7 The risk of Guillain-Barré syndrome following infection with Campylobacter jejuni; McCarthy N et al.; To estimate the incidence of Guillain-Barre syndrome (GBS) following Campylobacter jejuni infection (CI) we studied three populations where outbreaks of CI had occurred involving an estimated 8000 cases . No case of GBS was detected in the 6 months following the outbreaks in the local populations . The point estimate for the risk of GBS following CI estimated in this study was 0 in 8000 (95% confidence interval 0-3). Commun Dis Intell, 1999 Jan 21, 23(1), 1 - 27 Australia's notifiable diseases status, 1997 . Annual report of the National Notifiable Diseases Surveillance System; O'Brien E et al.; In 1997 there were 89,579 notifications to the National Notifiable Diseases Surveillance System . A notable feature of 1997 was the pertussis outbreak which peaked towards the end of the year and resulted in 10,668 cases being notified . The highest number of notifications received was for hepatitis C (unspecified) with 19,692 notifications; this is the first year for which data have been reported for New South Wales and South Australia for this disease category . The number of measles cases rose after the low number reported in 1996 but is still well below the number reported in the outbreak years of 1993 and 1994 . Rubella notifications continued to decline in 1997 . Notifications of Haemophilus influenzae type b appeared to have stabilised at a low rate, having declined markedly after introduction of the conjugated vaccine in 1992 . The number of cases of campylobacteriosis remained steady after having risen for several years . Notifications of hepatitis A cases rose considerably, much of this being due to one outbreak in New South Wales . The number of cases of salmonellosis rose while shigellosis numbers dropped slightly . Notifications for chlamydial infection and gonococcal infection continued to rise, whilst those for syphilis continued to fall. J Commun Dis, 1998 Sep, 30(3), 159 - 62 Isolation of nalidixic acid resistant Campylobacters from cases of paediatric diarrhoea in Chennai; Ananthan S et al.; A short term investigation on the Campylobacter enteritis among children under 10 years of age was carried out in Chennai . The study revealed an isolation rate of 11 per cent in 100 patients suffering from acute diarrhoea comprising C . jejuni (8%) and C . coli . (3%) . Among the two culture methods used, the candle jar method was found to be superior to plastic bag incubation system in recovering campylobacters on charcoal cefeperazone deoxycholate agar . While all the isolates were sensitive to ciprofloxacin, all of them exhibited resistance to nalidixic acid. Diagn Microbiol Infect Dis, 1999 Mar, 33(3), 181 - 6 Spectrum and antimicrobial activity of alexomycin (PNU-82, 127), a peptide compound projected for use in animal health; Marshall SA et al.; Alexomycin (PNU-82, 127) is a thiopeptide antimicrobial complex intended for veterinary practice that belongs to a series of cyclic peptides produced by Streptomyces arginensis . MICs against selected routine and fastidious clinical isolates of animal and human origin were determined by broth microdilution or agar dilution reference methods . Alexomycin was active against Gram-positive pathogens such as oxacillin-susceptible and -resistant Staphylococcus aureus and coagulase-negative staphylococci (260 strains; MIC90, 0.5 microgram/mL), as well as Enterococcus species (95 strains; MIC90, 0.25 to 0.5 microgram/mL), and generally inactive against Gram-negative aerobes . Alexomycin had more potent activity against Streptococcus bovis (MIC90, 0.12 microgram/mL), S . agalactiae (MIC90, 0.12 microgram/mL), Corynebacterium species (MIC90, 0.06-0.12 microgram/mL), and Listeria monocytogenes (MIC90, 0.5 microgram/mL) . Alexomycin activity was limited against Bacillus species (MIC90, 1 microgram/mL), Neisseria meningiditis (MIC90, 2 micrograms/mL), Haemophilus influenzae (MIC90, 8 micrograms/mL), Moraxella catarrhalis (MIC90, 16 micrograms/mL), and Campylobacter jejuni (MIC90, 32 micrograms/mL) . This thiopeptide complex was also found to be stable at low concentrations (0.015-32 micrograms/mL) in Mueller-Hinton broth for up to 24 h, possesses static antimicrobial activity and did not produce resistant mutants after multiple passages at subinhibitory drug concentrations . Alexomycin seems to have potential for use as a feed additive to increase feed efficiency and promote growth in poultry and swine as well as other applications against Gram-positive pathogens. Acta Crystallogr D Biol Crystallogr, 1999 Jan, 55 ( Pt 1), 299 - 301 Epub 1999 Jan 01. Expression, purification, crystallization and preliminary X-ray diffraction results from Campylobacter jejuni ferritin; Clerte S et al.; The prokaryotic ferritin gene of Campylobacter jejuni was overexpressed in Escherichia coli under control of the bacteriophage T7 promoter and the protein (Cj-FTN) purified . Preliminary crystallization experiments have been performed using the hanging-drop vapour-diffusion method with ammonium sulfate as the precipitant . Diffraction studies show the crystals belong to the I432 space group (a = 151.52 A) . Structure solution by molecular replacement is in progress while crystal quality improvement is carried out. Plasmid, 1999 Mar, 41(2), 97 - 109 Molecular characterization and interstrain variability of pHPS1, a plasmid isolated from the Sydney strain (SS1) of Helicobacter pylori; De Ungria MC et al.; The 5846-bp circular plasmid pHPS1 of Helicobacter pylori Sydney strain, SS1, was cloned, sequenced, and structurally characterized . The SS1 strain is widely used in animal studies of H . pylori infection . The sequence of pHPS1 revealed three open reading frames (ORFs), all of which are transcribed . Two ORFs encode putative plasmid replication proteins, RepA and RepB, similar to replicases resident on theta plasmids . In contrast, the function of ORF2 remains cryptic due to the absence of sequence similarity with any known protein in sequence databases . In addition, species specificity of these three coding regions was shown using DNA dot blot hybridization in 57 diverse clinical H . pylori isolates and 32 Helicobacter and Campylobacter strains . RepA appears to be the predominant plasmid replication protein of H . pylori and the deduced amino acid sequence was highly conserved (76-96%) in 8 H . pylori isolates, including SS1 . RepB was detected in 3 H . pylori isolates examined in this study, 2 of which possess only the repB gene . Analysis of the protein sequences of these two replicases, together with previously characterized H . pylori plasmid replication proteins, supports the formation of a distinct class of H . pylori plasmid proteins . Moreover, comprehensive analysis of the whole genome sequence of H . pylori strain 26695, pHPS1, and other H . pylori plasmid sequences that are available revealed interesting insights as to the occurrence of plasmid-mediated recombination within H . pylori . Common regions between plasmids and chromosome sequences of H . pylori were identified in this study which could only have arisen by genetic recombination, thus providing the first line of evidence, albeit indirectly, of the contribution of H . pylori plasmids in generating an extensive genetic heterogeneity characteristic of this important gastroduodenal pathogen . J Periodontal Res, 1999 Jan, 34(1), 25 - 33 Clinical and microbiological characteristics of smokers with early onset periodontitis; Kamma JJ et al.; Cigarette smoking is a potential risk factor which has recently been associated with periodontal disease progression . The objective of this study was to compare the microbial profile of smokers and non-smokers in a group of patients with early onset periodontitis . The study population consisted of 60 healthy individuals, 40 males and 20 females aged 22 to 35 yr, exhibiting early onset periodontitis . Thirty patients were smokers (30.9 cigarettes/d) and 30 non-smokers . Smokers had a higher proportion of deep pockets (PD >5 mm), especially in the maxilla anterior and premolar regions (p < 0.001) and presented a significantly greater mean probing depth and attachment loss (p <0.05) in diseased sites and a significantly greater alveolar bone loss (p <0.01) compared to non-smokers . Two pooled bacterial samples were obtained from each patient . Samples were collected from the deepest periodontal pockets of each quadrant . The samples were cultured anaerobically and in 10% CO2 plus air for bacterial isolation using selective and non-selective media . Isolates were characterized to species level by conventional biochemical tests and various identification kits . Smokers harboured a greater number of bacteria in total . Analysis of bacterial counts using the ANOVA (Mann-Whitney U-test) showed that Staphylococcus aureus, Peptostreptococcus micros, Campylobacter concisus, Escherichia coli, Bacteroides forsythus, C . gracilis, C . rectus, Porphyromonas gingivalis, Selenomonas sputigena, Candida albicans and Aspergillus fumigatus were found in significantly higher numbers and more frequently in smokers while Streptococcus intermedius, A . naeslundii, A . israelii and Eubacterium lentum were detected more frequently and in significantly higher proportions in non-smokers . The isolation of bacteria belonging to the exogenous flora such as E . coli, C . albicans, A . fumigatus and S . aureus in smokers' microbiota underscores the importance of the host that is adversely affected by cigarette smoking. An Esp Pediatr, 1999 Jan, 50(1), 25 - 8 {Lumbar puncture in a pediatric emergency department: something more than a diagnostic technic}; Mintegui Raso S et al.; OBJECTIVE: The aim of this study was to know the incidence of serious bacterial infections (SBI) in children without sepsis or intracranial infection in which spinal puncture (LP) was performed in an Emergency Department . PATIENTS AND METHODS: A retrospective study of all 471 previously healthy children between 1 month and 14 years of age in which a lumbar puncture was performed between July 1995 and March 1997 in the Emergency Department of our hospital was performed . RESULTS: Two hundred and three children (43%) had sepsis, meningitis or encephalitis (aseptic meningitis 149, 31.6%; sepsis-bacterial meningitis 26, 5.5%; nonspecific meningitis 26, 5.5%; encephalitis 2, 0.4%) and 14 (5.2%) had pneumonia . Of the other 254 children, 36 (14.1%) had a SBI: 19 urinary tract infections (E . coli), 11 bacteremia (Streptococcus pneumoniae 8, Salmonella enteritidis 1, Proteus mirabilis 1, E . coli 1, the latter two also having a positive urine culture) and 6 bacterial gastroenteritis (salmonella 5, Campylobacter jejuni 1) . The incidence of SBI was significantly higher in the group of children younger than 5 years old (32/175, 18.2%) than in the older group (4/79, 5.0%, p = 0.009) . Two patients died (one with pneumococcal meningitis and one with meningococcal sepsis) . CONCLUSIONS: Children with fever and a normal result in the