J Clin Microbiol, 2000 Oct, 38(10), 3800 - 10 Evaluation of phenotypic and genotypic methods for subtyping Campylobacter jejuni isolates from humans, poultry, and cattle; Nielsen EM et al.; Six methods for subtyping of Campylobacter jejuni were compared and evaluated with a collection of 90 isolates from poultry, cattle, and sporadic human clinical cases as well as from a waterborne outbreak . The applied methods were Penner heat-stable serotyping; automated ribotyping (RiboPrinting); random amplified polymorphic DNA typing (RAPD); pulsed-field gel electrophoresis (PFGE); restriction fragment length polymorphisms of the flagellin gene, flaA (fla-RFLP); and denaturing gradient gel electrophoresis of flaA (fla-DGGE) . The methods were evaluated and compared on the basis of their abilities to identify isolates from one outbreak and discriminate between unrelated isolates and the agreement between methods in identifying clonal lines . All methods identified the outbreak strain . For a collection of 80 supposedly unrelated isolates, RAPD and PFGE were the most discriminatory methods, followed by fla-RFLP and RiboPrinting . fla-DGGE and serotyping were the least discriminative . All isolates included in this study were found to be typeable by each of the methods . Thirteen groups of potentially related isolates could be identified using a criterion that at least four of the methods agreed on clustering of isolates . None of the subtypes could be related to only one source; rather, these groups represented isolates from different sources . Furthermore, in two cases isolates from cattle and human patients were found to be identical according to all six methods. J Clin Microbiol, 2000 Oct, 38(10), 3550 - 4 Diarrheagenic Escherichia coli and acute and persistent diarrhea in returned travelers; Schultsz C et al.; To determine the role of diarrheagenic Escherichia coli in acute and persistent diarrhea in returned travelers, a case control study was performed . Enterotoxigenic E . coli (ETEC) was detected in stool samples from 18 (10.7%) of 169 patients and 4 (3.7%) of 108 controls . Enteroaggregative E . coli (EAggEC) was detected in 16 (9.5%) patients and 7 (6.5%) controls . Diffuse adherent E . coli strains were commonly present in both patients (13%) and controls (13.9) . Campylobacter and Shigella species were the other bacterial enteropathogens most commonly isolated (10% of patients, 2% of controls) . Multivariate analysis showed that the presence of ETEC was associated with acute diarrhea (odds ratio {OR}, 6.7; 95% confidence interval {CI}, 1.5 to 29.1; P = 0.005), but not with persistent diarrhea (OR, 1.6; 95% CI, 0.4 to 7.4) . EAggEC was significantly more often present in patients with acute diarrhea than in controls (P = 0.009), but no significant association remained after multivariate analysis . ETEC and EAggEC are frequently detected in returned travelers with diarrhea . The presence of ETEC strains is associated with acute but not with persistent diarrhea. J Health Popul Nutr, 2000 Jun, 18(1), 33 - 8 Isolation and antibiotic susceptibility of Salmonella, Shigella, and Campylobacter from acute enteric infections in Egypt; Wasfy MO et al.; While Campylobacter, Salmonella, and Shigella remain major contributors to acute enteric infections, few studies on these pathogens have been conducted in Egypt . From January 1986 to December 1993, 869 Salmonella, Shigella and Campylobacter strains were isolated from stool specimens from 6,278 patients, presenting to the Abbassia Fever Hospital, Cairo, Egypt, with acute enteric infections . Salmonella predominated, totalling 465 isolates, followed by Shigella with 258 isolates, and Campylobacter with 146 isolates . Of the Shigella isolates, 124 were Shigella flexneri, 49 were S . sonnei, 47 were S . dysenteriae (mainly serotype 1, 2, and 3), and 38 were S . boydii . Campylobacter spp . comprised 92 Campylobacter jejuni and 54 C . coli isolates . Isolation of Salmonella was highest during the months of February-March, June-July, and October-November, while that of Shigella was maximal from July to October . Isolation of Campylobacter increased during May-June and again during August-October . Although Salmonella was sensitive to amikacin, aztreonam, ceftriaxone, and nalidixic acid, it was, however, resistant to erythromycin, streptomycin, ampicillin, chloramphenicol, and tetracycline . Shigella (> 80%) was sensitive to amikacin, ceftriaxone, cephalothin, sulphamethoxazole-trimethoprim (except S . sonnei), aztreonam, and nalidixic acid . Resistance (> 50%) was noted only for ampicillin, chloramphenicol, and tetracycline . C . jejuni and C . coli were resistant to cephalothin, aztreonam, and streptomycin . Some of the above antibiotics were employed to characterize the Egyptian isolates, but did not have any clinical utility in the treatment of diarrhoea . Significant differences (p < 0.05) were observed in the resistance profiles of Shigella and Salmonella between late 1980s and early 1990s . The results suggest the use of fluoroquinolones or a third-generation cephalosporin as an empirical treatment of enteric diseases . However, alternative control strategies, including the aggressive development of broadly protective vaccines, may be more effective approaches to curbing morbidity and mortality due to acute enteric infections. J Health Popul Nutr, 2000 Jun, 18(1), 23 - 6 Identification of enteric pathogens in HIV-positive patients with diarrhoea in northern India; Prasad KN et al.; Enteric pathogens associated with chronic diarrhoea in HIV-positive patients were studied . The study was conducted during January 1995-December 1998 . Stool specimens from all diarrhoea patients (n = 26) were examined microscopically for ova and parasites using wet preparations and stained smears . Stool samples from diarrhoea patients were also cultured on appropriate media to isolate enteric bacterial pathogens . Of the 59 patients, 26 (44%) had prolonged diarrhoea for more than 4 weeks . Enteric pathogens were detected in 19 (73%) of the 26 patients: 17 patients harboured a single pathogen, and 2 patients had mixed pathogens . The detection rate of emerging parasites, including Isospora, Cryptosporidium, Blastocystis hominis, and Strongyloides stercoralis as a single agent, was significantly higher than conventional pathogens (50% vs 19.2%; p < 0.05) . Only one patient harboured both conventional and emerging pathogens (Entamoeba histolytica and Cryptosporidium) . Isospora belli was detected in 8 (31%) of the 26 diarrhoea patients: in 7 (27%) patients as a single agent and in one patient with S . stercoralis . Cryptosporidium was identified in 3 (11%) diarrhoea patients: in 2 (8%) patients as a single agent and in one patient with E . histolytica, followed by B . hominis in 2 (8%) patients . E . histolytica was most commonly isolated (3/26; 11.5%), followed by Giardia lamblia, enteropathogenic Escherichia coli, and Campylobacter jejuni (one patient each) . Parasitic pathogens were frequently associated with HIV-positive patients with diarrhoea in northern India . I . belli was the most frequent parasite isolated, followed by Cryptosporidium . Stools of all HIV-positive patients with diarrhoea should thoroughly be investigated to identify aetiologic agents for proper management. Brain, 2000 Oct, 123 ( Pt 10), 2171 - 8 Haemophilus influenzae infection and Guillain-Barré syndrome; Mori M et al.; It has been reported recently that Haemophilus influenzae can elicit an axonal form of Guillain-Barre syndrome . To investigate the incidence and features of H . influenzae-related Guillain-Barre syndrome, anti-H . influenzae antibody titres were measured by enzyme-linked immunosorbent assay (ELISA) in 46 consecutive Japanese patients with Guillain-Barre syndrome, 49 normal controls, 24 patients with multiple sclerosis and 27 patients with amyotrophic lateral sclerosis (ALS) . Whole bacteria of non-encapsulated (non-typable) H . influenzae isolated from one of the Guillain-Barre syndrome patients was the antigen used . Elevated anti-H . influenzae antibodies for two or three classes of IgG, IgM and IgA were found in six (13%) Guillain-Barre syndrome patients, but not in the normal controls and patients with multiple sclerosis or ALS . The incidence was significantly higher in patients with Guillain-Barre syndrome than in the normal controls (P = 0.01) and patients with multiple sclerosis or ALS (P = 0.009) . Western blot analysis confirmed that the H . influenzae-positive patients' IgG recognized the lipopolysaccharides of H . influenzae . Guillain-Barre syndrome patients with anti-H . influenzae antibodies showed relatively uniform clinical and laboratory features: prodromal respiratory infection, less frequent cranial and sensory nerve involvement, pure motor axonal degeneration on electrophysiology, and positivity for IgG anti-GM1 antibodies . Although the features were similar to those in Guillain-Barre syndrome patients infected by Campylobacter jejuni, the recoveries seemed to be better in patients with H . influenzae-related Guillain-Barre syndrome . It is concluded that a form of Guillain-Barre syndrome occurs after respiratory infection by H . influenzae in the Japanese population . A particular strain of non-typable H . influenzae has a ganglioside GM1-like structure and elicits axonal Guillain-Barre syndrome similar to C . jejuni-related Guillain-Barre syndrome. No To Shinkei, 2000 Aug, 52(8), 715 - 7 {A case report of Bickerstaff's brainstem encephalitis with positive anti GQ 1 b, GT 1 a, GM 1 ganglioside antibodies}; Nemoto H et al.; Patient was an 18-year-old female student . After she had symptoms of common cold for 3 days, she developed somnolence, diplopia, dysarthria, urinary disturbance and ataxia . On admission neurological examination revealed coma with mydriasis, ophthalmoplegia, ptosis and weakness of the upper limbs . Light reflex, corneal reflex and oculocephalic test were all negative . Deep tendon reflexes were brisk and extensor toe signs were positive bilaterally . She did not have nuchal rigidity . Laboratory test revealed normal cerebrospinal fluid with negative myelin basic protein . Brain MRI, brainstem evoked potentials presented no abnormality . EMG revealed normal conduction velocity and no conduction block . EEG had diffuse theta and delta slowing . Culture of the stool represented no Campylobacter jejuni . At the fifth day of admission consciousness level improved, and other neurological findings disappeared in about 6 weeks . She had anti GQ 1 b, GT 1 a(IgG, IgM) and anti GM 1(IgM) antibodies in the serum . We made a diagnosis of Bickerstaff's brainstem encephalitis from these neurological symptoms and clinical course . The main lesion was present in the brainstem from midbrain to medulla oblongata in the midline . High titer of anti GT 1a antibody may be related to the ophthalmoplegia as noted in Miller Fisher syndrome . As a result of EMG and stool culture, it denied the complication of Guillain-Barre syndrome . We had no proof of the reason of the presentation of anti GM 1 antibody. Eur J Radiol, 2000 Sep, 35(3), 154 - 67 The epidemiology and the pathogenesis of inflammatory bowel disease; Karlinger K et al.; The etiology of inflammatory bowel disease (IBD) is still unknown . However, a satisfactory solution cannot be far away . IBD actually encompasses two diseases, i.e . Crohn's disease (CD) and ulcerous colitis (UC) . These diseases resemble each other so closely that they cannot be distinguished even pathologically, but differ from each other sufficiently to regard them as independent entities . Epidemiological observations may be helpful in identifying the true causative factors of this evasive disease . Geographically, the prevalence of the disease has a slope from North to South and, to a lesser degree, from West to East . The Western-Eastern discrepancy can be attributed to a difference in Western life styles . The incidence of the disease has been increasing world-wide of late, but its spread has been slowing down in highly affected countries . Racial and ethnic relations in different populations and immigration studies offer interesting data which can reflect genetic, inherited, environmental and behavioural factors . The disease seems to have a characteristic racial-ethnic distribution: the Jewish population is highly susceptible everywhere, but its prevalence in that population nears that of the domestic society in which they live . In Hungary, the Roma (Gypsies) have a considerably lower prevalence than the average population . This can be attributed to a genetic or environmental influence . According to age, the onset of the disease occurs more often in the second or the third decade of life, but there also is another peak in the 60s . Regarding sexual distribution, there is a slight preponderance of colitis ulcerosa in men and of Crohn's disease in women . It may correspond to the stronger auto-immune affection in the process of Crohn's disease . Environmental factors and behavioural influences also are investigated . Diet, the role of the early ages, smoking habits and the influence of hormonal status and drugs are viewed as useful contributing factors in the manifestation of the disease . Genetic studies show that one-fourth of IBD patients have an affected family member . HLAB27 histocombatibility also plays an important, but not determining role in the development of the disease . Genetic factors seem to have a stronger influence in Crohn's disease than ulcerative colitis . The existence of multiple sclerosis-IBD families may reflect the common genetic background or the similar microbial effect as well . A great number of bacterial and viral factors has been suspected of being infectious factors in IBD, mostly in CD . Mycobacteria, Yersinia, Campylobacter, Clostridium, Clamidias, etc . as well as bacteria and some viruses such as herpes and rotavirus and the primary measles virus . None of them has been proven as a real and exclusively pathogenic factor . Immunological background has an important function in the manifestation of the disease . If an individual has a genetic susceptibility to infections, the down regulation of an inflammation in the bowel wall does not occur in a proper way . This initiates the auto-immune process which is a self-increasing cycle . Extra-intestinal manifestations of IBD are of high importance because they can not only follow intestinal symptoms, but precede them by years . Hepatic and biliary disturbances (primary sclerosing cholangitis), are the most serious complications . Mucocutaneous manifestations can be the first appearance of the main disease (in the mouth) . Auto-immune consequences (erythema nodosum) or complications caused even by the therapy can occur . Ocular and musculoskeletal manifestations supposedly have the same genetic background and often precede the intestinal symptoms . Considering the epidemiological, genetic and immunological data, we can conclude that ulcerative colitis and Crohn's disease are heterogeneous disorders of mutifactorial etiology in which hereditary (genetic) and environmental (microbial, behaviour) factors interact to produce the disease. Berl Munch Tierarztl Wochenschr, 2000 Jul-Aug, 113(7-8), 289 - 94 {Peracute mortality in common cranes (Grus grus)}; Prange H et al.; Out of a nonbreeding group of cranes, 10 birds died peracutely at the end of April 1998 . The pathological investigation showed changes in the intestine, liver and kidneys caused probably by an intoxication; but corresponding analyses did not result in a specified poison . The proof of E . coli, Cl . perfringens and Campylobacter jejuni is to be interpreted as a subordinate result . 7 of 8 cranes had a low to high infestation with endoparasites (Porrocaeum spp., Eimeria pusilla, Echinostoma spp.) . 5 of 8 birds showed leaness, possibly as a result of the migration exertion . Further on, the analysis results of a 9th crane found at another place are included in this paper. Infect Immun, 2000 Oct, 68(10), 5679 - 89 Sequence polymorphism, predicted secondary structures, and surface-exposed conformational epitopes of Campylobacter major outer membrane protein; Zhang Q et al.; The major outer membrane protein (MOMP), a putative porin and a multifunction surface protein of Campylobacter jejuni, may play an important role in the adaptation of the organism to various host environments . To begin to dissect the biological functions and antigenic features of this protein, the gene (designated cmp) encoding MOMP was identified and characterized from 22 strains of C . jejuni and one strain of C . coli . It was shown that the single-copy cmp locus encoded a protein with characteristics of bacterial outer membrane proteins . Prediction from deduced amino acid sequences suggested that each MOMP subunit consisted of 18 beta-strands connected by short periplasmic turns and long irregular external loops . Alignment of the amino acid sequences of MOMP from different strains indicated that there were seven localized variable regions dispersed among highly conserved sequences . The variable regions were located in the putative external loop structures, while the predicted beta-strands were formed by conserved sequences . The sequence homology of cmp appeared to reflect the phylogenetic proximity of C . jejuni strains, since strains with identical cmp sequences had indistinguishable or closely related macrorestriction fragment patterns . Using recombinant MOMP and antibodies recognizing linear or conformational epitopes of the protein, it was demonstrated that the surface-exposed epitopes of MOMP were predominantly conformational in nature . These findings are instrumental in the design of MOMP-based diagnostic tools and vaccines. Infect Immun, 2000 Oct, 68(10), 5663 - 7 Campylobacter fetus sap inversion occurs in the absence of RecA function; Ray KC et al.; Phase variation of Campylobacter fetus surface layer proteins (SLPs) occurs by inversion of a 6.2-kb DNA segment containing the unique sap promoter, permitting expression of a single SLP-encoding gene . Previous work has shown that the C . fetus sap inversion system is RecA dependent . When we challenged a pregnant ewe with a recA mutant of wild-type C . fetus (strain 97-211) that expressed the 97-kDa SLP, 15 of the 16 ovine-passaged isolates expressed the 97-kDa protein . However, one strain (97-209) expressed a 127-kDa SLP, suggesting that chromosomal rearrangement may have occurred to enable SLP switching . Lack of RecA function in strains 97-211 and 97-209 was confirmed by their sensitivity to the DNA-damaging agent methyl methanesulfonate . Southern hybridization and PCR of these strains indicated that the aphA insertion into recA was stably present . However, Southern hybridizations demonstrated that in strain 97-209 inversion had occurred in the sap locus . PCR data confirmed inversion of the 6.2-kb DNA element and indicated that in these recA mutants the sap inversion frequency is reduced by 2 to 3 log(10) units compared to that in the wild type . Thus, although the major sap inversion pathway in C . fetus is RecA dependent, alternative lower-frequency, RecA-independent inversion mechanisms exist. J Neurol Neurosurg Psychiatry, 2000 Oct, 69(4), 522 - 4 Unusual T cell receptor phenotype V gene usage of gamma delta T cells in a line derived from the peripheral nerve of a patient with Guillain-Barré syndrome; Cooper JC et al.; Guillain-Barre syndrome is considered to be an immune mediated disorder but the relative role of T cells and antibodies in its pathogenesis is unclear . As gut infection with Campylobacter jejuni is the most common antecedent infection it is possible that gut derived T lymphocytes might play a part in the development of the syndrome.The T cell receptor phenotype (TCR) of a nerve gamma delta T cell line obtained from a sural nerve biopsy taken from a patient with a demyelinating form of GBS was determined using polymerase chain reaction (PCR) and flow cytometry (FACS) . This TCR was compared with the phenotype preferentially expressed in the peripheral blood of the same patient.The T cell nerve line was found to express V gamma 8/delta 1 which represents an unusual T cell subset normally found on lymphocytes resident in epithelial tissue such as the gut . The peripheral blood gamma delta T lymphocytes from the patient were of the V gamma 9/delta 2 subset, which is the phenotype predominantly expressed in the peripheral blood of healthy subjects . In conclusion, the presence of this unusual population of V gamma 8/delta 1(+) lymphocytes in nerve would be consistent with a pathogenetic role for gut associated lymphocytes in the pathogenesis of Guillain-Barre syndrome associated with C jejuni. Microb Drug Resist, 2000 Summer, 6(2), 91 - 8 An integron cassette carrying dfr1 with 90-bp repeat sequences located on the chromosome of trimethoprim-resistant isolates of Campylobacter jejuni; Gibreel A et al.; The frequent occurrence of high-level trimethoprim resistance in clinical isolates of Campylobacter jejuni was shown to be related to the acquisition of foreign resistance genes (dfrl or dfr9 or both) coding for resistant variants of the enzyme dihydrofolate reductase, the target of trimethoprim . The dfr1 gene detected on the chromosome of 40 different clinical strains of C . jejuni was studied further regarding structure and genetic organization . Most of the dfr1 genes were found as integron cassettes inserted in the chromosome . In 36% of the examined isolated, the dfr1 gene showed identity to that previously characterized in trimethoprim-resistant Escherichia coli . In 40% of the cases, however, a variant of the dfr1 gene containing a 90-bp direct repeat was detected, and in 5% of the isolates, the repeat-containing dfr1 variant was found to occur in the form of two cassettes in tandem in an integron context . The existence of the 90-bp repeat within the coding sequence of the dfr1gene was found to play a role in the adaptation of C . jejuni to ambient concentrations of trimethoprim. Exp Toxicol Pathol, 2000 Aug, 52(4), 287 - 96 Susceptibility to secondary bacterial infections in growing pigs as an early response in ochratoxicosis; Stoev SD et al.; Mycotoxic nephropathy was induced in twelve 14 kg pigs fed a dietary component, moulded by Aspergillus ochraceus and contributing ochratoxin A at 1 or 3 ppm for up to 3 weeks . Concurrently, salmonellosis arose spontaneously in all six animals treated at 3 ppm and all died between days 15 and 17 . Two of the six pigs in the 1 ppm group died similarly but the rest, and all of six control animals, were unaffected . Clinical biochemistry and histology revealed changes typical of renal ochratoxicosis in all ochratoxin-treated pigs . Clinical and pathomorphological changes typical of salmonellosis were evident in all those that died and Salmonella choleraesuis was consistently isolated from their faeces and liver . In a further experiment at 1 ppm ochratoxin A in animals immunised against S . choleraesuis haemorrhagic diarrhoea resulted instead, associated with Serpulina hyodysenteriae and Campylobacter coli . There was concomitant evidence of immunosuppression and delayed response to immunization . For the first time, susceptibility to natural infectious disease has been demonstrated in pigs exposed to the immunotoxicity of ochratoxin A . Differentiation of biochemical and histological changes attributable to ochratoxicosis or to secondary disease may require reinterpretation of a classical description of experimental porcine ochratoxicosis. J Periodontal Res, 2000 Aug, 35(4), 232 - 41 Prevalence of periodontal pathogens in localized and generalized forms of early-onset periodontitis; Mullally BH et al.; The primary objectives of this study were to investigate the prevalence of 8 putative periodontal pathogens in subjects with early-onset periodontitis (EOP) and to evaluate the microbial differences between localized and generalized forms of this periodontal disease condition . Thirty-one females and 11 males with a mean age of 30.3 (s.d . 4.0) years were examined . Seventeen subjects had generalized (GEOP) and 25 had localized early-onset periodontitis (LEOP) . Subgingival plaque samples were assayed using PCR which provided subject prevalence data for the pathogens; Bacteroides forsythus 78.6%, Treponema denticola 88.1%, Actinobacillus actinomycetemcomitans 19.0%, Porphyromonas gingivalis 16.7%, Prevotella intermedia 40.4%, Prevotella nigrescens 61.9%, Eikenella corrodens 42.3% and Campylobacter rectus 92.8% . Only 3 healthy sites harbored one or more of these periodontal pathogens . Seven of the 8 subjects positive for A . actinomycetemcomitans had LEOP . P . intermedia was present in 58.8% of GEOP compared with 28% of LEOP subjects (p=0.046) . At 82.4% of GEOP sites P . nigrescens was present while this bacteria was detected at 52% of LEOP (p=0.044) . P . gingivalis was isolated from 22.6% of females but no male subjects (p=0.084) . C . rectus was recovered from all female subjects compared to 72.7% of males (p=0.014) . A . actinomycetemcomitans (37.5%) and C . rectus (86.5%) were more frequently identified in non-smokers compared to 7.6% and 68.8% of smokers, respectively (p <0.05) . Microbial associations coincided with the clinical division of the cases into LEOP and GEOP in 83% of the subjects. J Clin Periodontol, 2000 Sep, 27(9), 648 - 57 Comparison of the microbiota of supra- and subgingival plaque in health and periodontitis; Ximenez-Fyvie LA et al.; BACKGROUND, AIMS: The purpose of the present investigation was to compare the microbial composition of supra and subgingival plaque in 22 periodontally healthy (mean age 32+/-16 years) and 23 adult periodontitis subjects (mean age 51+/-14 years) . METHODS: A total of 2358 supra and separately subgingival plaque samples were collected from the mesial aspect of all teeth excluding 3rd molars in each subject . Samples were examined for the presence and levels of 40 bacterial taxa using whole genomic DNA probes and checkerboard DNA-DNA hybridization . Clinical assessments including dichotomous measures of gingival redness, bleeding on probing, plaque accumulation and suppuration, as well as duplicate measures of pocket depth and attachment level, were made at 6 sites per tooth . Mean counts (x10(5), % DNA probe count and % sites colonized for each species were determined separately for supra and subgingival samples in each subject and then averaged across subjects in the 2 clinical groups . Significance of differences between healthy and periodontitis subjects was determined using the Mann-Whitney test and adjusted for multiple comparisons . RESULTS: Mean total DNA probe counts (x10(5), +/-SEM) for healthy and periodontitis subjects in supragingival plaque were 72.1+/-11 and 132+/-17.5, respectively (p<0.01), and in subgingival plaque 22.1+/-6.6 and 100.3+/-18.4, (p<0.001) . Porphyromonas gingivalis, Bacteroides forsythus and Treponema denticola could be detected in supragingival plaque samples of both healthy and periodontitis subjects . Actinomyces species were the dominant taxa in both supra- and subgingival plaque from healthy and periodontitis subjects . 4 Actinomyces species accounted for 63.2%, of supragingival and 47.2% of subgingival plaque in healthy subjects and 48.% and 37.8% in periodontitis subjects respectively . Increased proportions of P . gingivalis, B . forsythus, and species of Prevotella, Fusobacterium, Campylobacter and Treponema were detected subgingivally in the periodontitis subjects . P . gingivalis, B . forsythus and T . denticola were significantly more prevalent in both supra- and subgingival plaque samples from periodontitis subjects . CONCLUSIONS: The main differences between supra and subgingival plaque as well as between health and disease were in the proportions and to some extent levels of Actinomyces, "orange" and "red" complex species. FEMS Microbiol Lett, 2000 Sep 1, 190(1), 1 - 7 Characterization of gyrA mutations associated with fluoroquinolone resistance in Campylobacter coli by DNA sequence analysis and MAMA PCR; Zirnstein G et al.; Increasing numbers of fluoroquinolone-resistant Campylobacter coli isolates received at the Minnesota State Public Health Laboratory and at the Centers for Disease Control and Prevention have been a cause for concern . The gyrA quinolone resistance-determining regions of several fluoroquinolone-resistant isolates were sequenced to examine the mechanism of resistance . Ciprofloxacin-resistant C . coli isolates examined by DNA sequencing had a Thr-86 to Ile (ACT-->ATT) gyrA mutation, leading to resistance to fluoroquinolone antibiotics . A mismatch amplification mutation assay polymerase chain reaction protocol was developed to detect this gyrA mutation. Am J Reprod Immunol, 2000 Jul, 44(1), 30 - 40 Endocrine-immune interaction: alteractions in immune function resulting from neonatal treatment with a GnRH antagonist and seasonality in male primates; Mann DR et al.; PROBLEM: The effect of neonatal gonadotropin releasing hormone (GnRH) antagonist (Ant) treatment and seasonality on immune system development and function was investigated in male primates . METHOD OF STUDY: Neonatal male rhesus monkeys and marmosets were treated with Ant, and its effect on immune system morphology, circulating lymphocyte subsets, and cell- and humorally-mediated immune responses was assessed during development . In adult rhesus monkeys, we correlated seasonal changes in immune function with circannual fluctuations in immunoactive hormones . RESULTS: In neonatal marmosets, Ant reduced the number of B cells and T cells in the thymic medulla and T cells in the periarterial lymphatic sheaths (PALS) of the spleen . Ant also altered the development of, but did not permanently impair, the proliferative index (PI) of blood lymphocytes to mitogens . In vitro treatment of control lymphocytes with GnRH analogues altered their response to these proliferative agents . In neonatal rhesus monkeys, Ant treatment increased the frequency of clinical problems, lowered circulating levels of lymphocytes, total T cells, CD8+ T cells and B cells, and altered the PI of lymphocytes to mitogens . As adults, the cell- and humorally-mediated immune responses remained impaired . We also documented seasonal fluctuations in the prevalence of diseases, circulating immune cells and immune function in rhesus monkeys . The number of cases of campylobacteriosis and shigellosis was lowest in the winter and highest in the spring . Circulating numbers of white blood cells (WBC) and neutrophils and the PI of lymphocytes to mitogens were higher in the winter than in the summer . Natural killer cell activity also varied with season . Cortisol and leptin secretion exhibited circannual rhythms, rising in concert with decreasing photoperiod and increasing testicular activity in the fall . Conversely, prolactin levels declined with decreasing photoperiod and then rose in the spring . CONCLUSION: Neonatal exposure of male primates to Ant appears to alter early postnatal programming of immune function . In the rhesus monkey, immune function shows seasonal fluctuations that may be driven by circannual changes in the secretion of immunoactive hormones. Microbiology, 2000 Sep, 146 ( Pt 9), 2283 - 90 Concerted evolution of duplicate fla genes in Campylobacter; Meinersmann RJ et al.; Campylobacters have two similar copies (flaA and flaB) of their flagellin gene . It has been hypothesized that the two copies can serve for antigenic phase variation . Analysis of polymorphisms within aligned multiple DNA sequences of the Campylobacter flagellin genes revealed high pairwise homoplasy indexes between flaB/flaB pairs that were not observed between any flaA/flaA pairings or flaA/flaB pairings . Thus it seems there are constraints on the sequence of flaB that distinguish it from flaA . Nevertheless, segments of the two genes that are highly variable between strains are conserved between the flaA and flaB copies of the genes within a strain . The patterns of synonymous and non-synonymous differences suggest that one segment of the flagellin sequence is under selective pressure at the amino acid sequence level . Another segment of the protein is maintained within a strain by conversion or recombination . Comparisons of strict consensus amino acid sequences did not reveal any motifs that are uniquely FlaA or FlaB, but there are differences between FlaA and FlaB in those amino acids available for post-translational modification . The observed pattern of concerted evolution of portions of a structural gene is an unusual finding in bacteria and should be searched for with other duplicated genes . Concerted evolution was unexpected for genes involved in phase variation since it minimizes the antigenic repertoire that can be expressed by a single clone in the face of the host immune response. Lett Appl Microbiol, 2000 Sep, 31(3), 209 - 12 The incidence and PCR detection of Campylobacter upsaliensis in dogs and cats; Steinhauserova I et al.; A study was made of the incidence of Campylobacter upsaliensis among dogs and cats suffering from acute or chronic diarrhoea; 225 dogs and cats were examined and 51 strains were identified, 16 (7%) of which were Camp . upsaliensis . When rectal swabs were taken from a control group of 126 dogs and cats without clinical symptoms, 19 Campylobacter spp . and four Camp . upsaliensis were identified . All the Camp . upsaliensis strains were isolated in dogs . The Campylobacter strains were identified on the basis of their biochemical characteristics and by PCR (polymerase chain reaction). Helicobacter, 2000 Sep, 5(3), 142 - 7 Differentiation of clinical Helicobacter pullorum isolates from related Helicobacter and Campylobacter species; Melito PL et al.; BACKGROUND: Helicobacter pullorum, first detected in the liver and intestinal contents of poultry, was defined as a new species in 1994 . This organism has since been isolated from humans with gastroenteritis . Phenotypic as well as genotypic methods have been used to identify H . pullorum associated with cases of human disease . MATERIALS AND METHODS: Clinical isolates were submitted for identification to the National Laboratory for Enteric Pathogens by Provincial Public Health Laboratories within Canada . Phenotypic characterization was conducted using a variety of growth and biochemical tests including oxidase, catalase, indoxyl acetate, H2S production in triple sugar iron (TSI) agar, antimicrobial susceptibility testing, and fatty acid analysis . Genotypic identification was performed using a polymerase chain reaction-restriction fragment-length polymorphism (PCR-RFLP) analysis of a 1-kb fragment of the Helicobacter 16S rRNA gene . RESULTS: During the last 7 years (1993-1999) a total of 11 isolates of H . pullorum were detected from patients with gastroenteritis for inclusion in this study . Typically, these isolates were oxidase and catalase positive, produced optimal growth at 42 degrees C, and produced H2S in TSI . Of these 11 isolates, 1 showed DNase activity, while another did not produce H2S in TSI, and only 2 showed tolerance to 1% bile . Antimicrobial susceptibility assays indicated that 6 of the 11 strains were resistant to nalidixic acid . The fatty acid profiles of the isolates were similar to each other and provided a distinguishing profile from the other related species . Genetically identical and distinct species-specific restriction fragment-length polymorphism (RFLP) patterns were produced using the restriction enzymes Bsr I and Dde I . CONCLUSION: Phenotypic and genotypic procedures were used to identify H . pullorum . Interspecies phenotypic variability was apparent and supported the use of a polyphasic approach for identification . Similarities to the more prominent human pathogens Campylobacter coli and C . lari were also noted . The use of a combination of phenotypic and, in particular, genotypic markers for H . pullorum should prove valuable both for epidemiological investigations and for the diagnosis of disease related to this emerging human pathogen. Mol Cell Probes, 2000 Aug, 14(4), 233 - 40 Rapid detection of Campylobacter fetus by polymerase chain reaction combined with non-radioactive hybridization using an oligonucleotide covalently bound to microwells; Casademont I et al.; Campylobacter fetus is recognized as a human and animal pathogen . The isolation and differentiation of C . fetus in diagnostic laboratories is hindered by its relatively slow growth and lack of distinguishing biochemical characteristics . We cloned and sequenced a 1581-bp DNA fragment, IG02, isolated from a C . fetus genomic library . This fragment was used as a probe on DNAs extracted from C . fetus strains and other Campylobacter species: IG02 hybridized only with DNAs from C . fetus strains . A PCR-based test was developed for the detection of C . fetus . A pair of oligonucleotide primers was designed to amplify a 141-bp fragment of IG02 . The amplified product was analysed by a non-radioactive sandwich hybridization in microtiter plate using a capture oligonucleotide and a biotin-labelled oligonucleotide for the detection . The combination of PCR and non-radioactive microplate hybridization is a convenient method for the rapid detection of C . fetus . J Clin Microbiol, 2000 Sep, 38(9), 3379 - 87 Comparative fingerprinting analysis of Campylobacter jejuni subsp . jejuni strains by amplified-fragment length polymorphism genotyping; Lindstedt BA et al.; Amplified-fragment length polymorphism (AFLP) analysis with the endonucleases BglII and MfeI was used to genotype 91 Campylobacter jejuni subsp . jejuni strains from outbreaks and sporadic cases . AFLP-generated fragments were labeled with fluorescent dye and separated by capillary electrophoresis . The software packages GeneScan and GelCompar II were used to calculate AFLP pattern similarities and to investigate phylogenetic relationships among the genotyped strains . The AFLP method was compared with two additional DNA-based typing methods, pulsed-field gel electrophoresis (PFGE) using SmaI and restriction fragment length polymorphism analysis on PCR products (PCR-RFLP) of the flaA and flaB genes . We found that AFLP analysis of C . jejuni strains is a rapid method that offers better discriminatory power than do both PFGE and PCR-RFLP . AFLP and, to a lesser extent, PCR-RFLP could differentiate strains within the same PFGE profiles, which also makes PCR-RFLP an alternative to PFGE . We were able to clearly distinguish 9 of 10 recognized outbreaks by AFLP and to identify similarities among outbreak and sporadic strains . Therefore, AFLP is suitable for epidemiological surveillance of C . jejuni and will be an excellent tool for source identification in outbreak situations. J Formos Med Assoc, 2000 Aug, 99(8), 612 - 7 Bacteremia due to Campylobacter species: high rate of resistance to macrolide and quinolone antibiotics; Lu PL et al.; BACKGROUND AND PURPOSE: Although the rate of isolation of Campylobacter from stool specimens in Taiwan is similar to those in other developed countries, Campylobacter bacteremia has rarely been reported in Taiwan, and the patterns of antimicrobial susceptibility of blood isolates to various antimicrobial agents remain unknown in the Taiwanese population . The purpose of this study was to determine the clinical characteristics of patients with Campylobacter infection in a university hospital in Taiwan and the antimicrobial susceptibility patterns of the Campylobacter isolates . METHODS: We retrospectively reviewed medical records of all patients with Campylobacter bacteremia treated in a university hospital between January 1991 and March 1999 . Minimum inhibitory concentrations of 13 antimicrobial agents to 10 stored blood isolates were determined using the E-test . RESULTS: Approximately half (52%) of the 21 patients had chronic liver disease and one-quarter had hepatobiliary or gastrointestinal malignancies . Thirteen (62%) patients had conditions that were associated with gastroenteritis . Other clinical manifestations associated with Campylobacter infection included cellulitis, perinatal sepsis, peritonitis, vascular catheter-related infection, and primary bacteremia . The duration of illness was generally short: approximately half (52%) of the 21 patients had fever lasting for only 1 day . Antimicrobial susceptibility testing of the 10 isolates revealed that most of the blood isolates were resistant to erythromycin and nalidixic acid (100% and 90%, respectively), while the rate of cross-resistance between erythromycin and azithromycin was 70%, and that between nalidixic acid and ciprofloxacin was 67% . CONCLUSIONS: Our observations suggest that Campylobacter bacteremia should be included in the differential diagnosis of patients with chronic liver disease or malignancies involving the hepatobiliary system or gastrointestinal tract who present with fever and gastroenteritis . Clinicians in Taiwan should be alert to the high rate of resistance of Campylobacter isolates to macrolide and quinolone antibiotics. Eur J Clin Microbiol Infect Dis, 2000 Jul, 19(7), 542 - 4 Outbreak of Campylobacter infection in a subartic community; Melby KK et al.; A presumably waterborne outbreak of Campylobacter jejuni/coli infection in a subarctic community is described . Drinking water supplied to residents was delivered unchlorinated during a 4-week period . No Campylobacter sp . was recovered from the water supply . Three hundred thirty individuals (15% of the 2,200 exposed) became ill . Diarrhoea, abdominal pain, fever, nausea and joint pain occurred in 81%, 30%, 29%, 43% and 21%, respectively . Nine percent reported swelling of joints, and two cases of reactive arthritis occurred . A Campylobacter sp . was isolated from 9 of 33 individuals who became ill and from 1 of 33 healthy controls . All culture-positive individuals, 46% of culture-negative ill persons and 27% of healthy controls were seropositive . All strains recovered had an identical DNA profile. Appl Environ Microbiol, 2000 Sep, 66(9), 4115 - 8 Quantitative immunocapture PCR assay for detection of Campylobacter jejuni in foods; Waller DF et al.; The rapid detection of food-borne bacterial pathogens as part of a quality control program is necessary for the maintenance of a safe food supply . In this report, we present our findings for an immunocapture PCR method for the detection of Campylobacter jejuni in foods . The method permits direct detection of the pathogen without an enrichment step and can be performed in approximately 8 h . Assay results are quantitative, and one cell in a milliliter sample can be detected . Application of the method to spiked milk samples and chicken skin washes did not affect the sensitivity of the assay. Appl Environ Microbiol, 2000 Sep, 66(9), 4029 - 36 Application of the 5'-nuclease PCR assay in evaluation and development of methods for quantitative detection of Campylobacter jejuni; Nogva HK et al.; Campylobacter jejuni is recognized as a leading human food-borne pathogen . Traditional diagnostic testing for C . jejuni is not reliable due to special growth requirements and the possibility that this bacterium can enter a viable but nonculturable state . Nucleic acid-based tests have emerged as a useful alternative to traditional enrichment testing . In this article, we present a 5'-nuclease PCR assay for quantitative detection of C . jejuni and describe its evaluation . A probe including positions 381121 to 381206 of the published C . jejuni strain NCTC 11168 genome sequence was identified . When this probe was applied, the assay was positive for all of the isolates of C . jejuni tested (32 isolates, including the type strain) and negative for all other Campylobacter spp . (11 species tested) and several other bacteria (41 species tested) . The total assay could be completed in 3 h with a detection limit of approximately 1 CFU . Quantification was linear over at least 6 log units . Quantitative detection methods are important for both research purposes and further development of C . jejuni detection methods . In this study, we used the assay to investigate to what extent the PCR signals generated by heat-killed bacteria interfere with the detection of viable C . jejuni after exposure at elevated temperatures for up to 5 days . An approach to the reduction of the PCR signal generated by dead bacteria was also investigated by employing externally added DNases to selectively inactivate free DNA and exposed DNA in heat-killed bacteria . The results indicated relatively good discrimination between exposed DNA from dead C . jejuni and protected DNA in living bacteria. Appl Environ Microbiol, 2000 Sep, 66(9), 3917 - 23 Amplified fragment length polymorphism analysis of Campylobacter jejuni strains isolated from chickens and from patients with gastroenteritis or Guillain-Barré or Miller Fisher syndrome; Duim B et al.; The high-resolution genotyping method of amplified fragment length polymorphism (AFLP) analysis was used to study the genetic relationships between Campylobacter jejuni strains infecting chickens (n = 54) and those causing gastroenteritis in humans (n = 53) . In addition, C . jejuni strains associated with the development of Guillain-Barre syndrome (GBS) (n = 14) and Miller Fisher syndrome (MFS) (n = 4), two related acute paralytic syndromes in human, were included . Strains were isolated between 1989 and 1998 in The Netherlands . The AFLP banding patterns were analyzed with correlation-based and band-based similarity coefficients and UPGMA (unweighted pair group method using average linkages) cluster analysis . All C . jejuni strains showed highly heterogeneous fingerprints, and no fingerprints exclusive for chicken strains or for human strains were obtained . All strains were separated in two distinct genetic groups . In group A the percentage of human strains was significantly higher and may be an indication that genotypes of this group are more frequently associated with human diseases . We conclude that C . jejuni from chickens cannot be distinguished from human strains and that GBS or MFS related strains do not belong to a distinct genetic group. J Periodontol, 2000 Jul, 71(7), 1144 - 50 Fusobacterium nucleatum involvement in adult periodontitis and possible modification of strain classification; Roques CG et al.; BACKGROUND: This investigation was designed to evaluate the involvement of Fusobacterium nucleatum clinical strains in adult periodontitis by subspecies and expression of hemagglutination activity . METHODS: Forty-nine Fusobacterium strains were isolated from 40 sites in 40 subjects presenting with adult periodontitis . F . nucleatum subspecies identification was based on the electrophoretic migration of glutamate dehydrogenase and 2-oxoglutarate reductase . Hemagglutination activity and inhibition by galactose were tested on sheep erythrocytes . RESULTS: The 49 isolates belonged to the F . nucleatum species with a predominance of the nucleatum (34.7%) followed by the vincentii (26.5%) subspecies . In parallel, 71% of the strains belonging to the nucleatum subspecies were preferentially associated with Porphyromonas gingivalis . Prevotella intermedia/nigrescens detection was essentially correlated with identification of Fusobacterium nucleatum subspecies vincentii . No correlation was established between any particular subspecies and the pathogenicity factors tested (hemagglutination and production of short-chain fatty acids) . On the other hand, significant predominance (65%, P= 0.017) of strongly hemagglutinating strains (titre > or =8 U) was observed in the sites where Porphyromonas gingivalis, Prevotella intermedia/nigrescens and/or Campylobacter rectus were not detected . These strains also showed higher butyric acid production . CONCLUSION: The importance of the adherence factors for Fusobacterium nucleatum strains and their multimodal aspect may indicate a higher pathogenicity or a higher involvement of certain strains and could lead to a classification of these strains, which is more closely related to their implication in the development of periodontal disease. J Peripher Nerv Syst, 1998, 3(1), 3 - 18 Anti-ganglioside antibody and neuropathy: review of our research; Yuki N; Some patients developed Guillain-Barre syndrome (GBS) after the administration of bovine gangliosides . Patients with GBS subsequent to Campylobacter jejuni enteritis frequently have IgG antibody to GM1 ganglioside . Fisher's syndrome (FS), a variant of GBS, is associated with IgG antibody to GQ1b ganglioside . We showed the existence of molecular mimicry between GM1 and lipopolysaccharide (LPS) of C . jejuni isolated from a GBS patient, and that between GQ1b and C . jejuni LPSs from FS patients . Several lines of evidence suggest a pathogenic role for anti-ganglioside antibodies . Some patients developed sensorimotor polyneuropathy after anti-GD2 antibody administration . Anti-GM1 antibody can block motor nerve conduction . The molecular mimicry between infectious agents and gangliosides may function in the production of anti-ganglioside antibodies and the development of GBS and FS . Anti-GQ1b IgG antibody is detected also in Bickerstaff's brainstem encephalitis and acute ophthalmoparesis, which suggests that these conditions are categorized as autoimmune diseases related to FS . Since a tryptophan-immobilized column effectively adsorb anti-GQ1b IgG antibody, immunoadsorption with the column should be considered as an alternative form of plasmapheresis for the anti-GQ1b IgG antibody syndrome. Microbios, 2000, 102(403), 159 - 64 Detection and investigation of Campylobacter jejuni by polymerase chain reaction-restriction fragment length polymorphism analysis; Kiuchi A et al.; A molecular typing approach for Campylobacter jejuni with restriction fragment length polymorphism (RFLP) analysis of the flagellin gene flaA in C . jejuni, was generated and studied . Using polymerase chain reaction (PCR)-RFLP with the restriction endonuclease Mbo I, it was demonstrated that C . jejuni could be divided into four types . Genotypic analysis of C . jejuni by PCR-RFLP is a valuable technique for epidemiological typing. Can Fam Physician, 2000 Aug, 46, 1634 - 8 Preparing patients to travel abroad safely . Part 4: Reducing risk of accidents, diarrhea, and sexually transmitted diseases; Thomas RE; OBJECTIVE: To present evidence-based recommendations on traveling abroad safely so family physicians can advise travelers on how to reduce risk of accidents, diarrhea, and sexually transmitted diseases (STDs) and how to treat diarrhea themselves if medical care is unavailable . QUALITY OF EVIDENCE: A MEDLINE search from 1990 to November 1998 found 163 articles on travel and accidents, 504 on travel and diarrhea, and 42 on travel and STDs . Titles and abstracts were reviewed, and randomized controlled trials (RCTs) and systematic reviews were sought . The Cochrane Collaboration database of systematic reviews and meta-analyses was searched for studies relevant to family physicians . MAIN MESSAGE: For preventing diarrhea, RCTs demonstrate that bismuth subsalicylate, doxycycline, ciprofloxacin, and trimethoprim-sulfamethoxazole are useful prophylactics . Once travelers have diarrhea, RCTs show that loperamide and zaldaride reduce symptoms and duration; quinolones, ciprofloxacin, norfloxacin, and oral aztreonam reduce abdominal symptoms and time to last liquid stool by several days; azithromycin is effective in treatment of ciprofloxacin-resistant Campylobacter, and trimethoprim-sulfamethoxazole is effective in treating cyclospora . There are no RCTs of preventing accidents and STDs abroad . Health Canada has issued a statement summarizing the risks of acquiring STDs abroad . CONCLUSION: Family physicians can advise their patients on how to reduce risk of travelers' diarrhea and how to treat it themselves on holiday . There is expert advice on how to reduce risk of STDs. Acta Neurol Scand, 2000 Aug, 102(2), 132 - 4 Is Campylobacter lipopolysaccharide bearing a GD3 epitope essential for the pathogenesis of Guillain-Barré syndrome? Yuki N, Koga M, Hirata K. The hypothesis has been proposed that the GD3 ganglioside-like lipopolysaccharide (LPS) is essential for and functions in the development of Guillain-Barre syndrome (GBS) and Miller Fisher syndrome (MFS) subsequent to Campylobacter jejuni enteritis . Our study showed that patients with GBS or MFS who had previously suffered diarrhea had anti-GD3 antibodies less often than those who had not had diarrhea . Sera from patients who showed GBS or MFS with the serologic evidence of prior C . jejuni infection had anti-GD3 antibodies less frequently than sera from those without evidence of infection . Statistical analysis showed that anti-GD3 antibodies were less frequent in patients with GBS or MFS from whom C . jejuni had been isolated than were other antiganglioside antibodies, such as anti-GM1 antibodies . These results could not support the above hypothesis. Infect Immun, 2000 Sep, 68(9), 5450 - 3 Random transposon mutagenesis of Campylobacter jejuni; Golden NJ et al.; Genetic studies of Campylobacter jejuni have been limited due to the lack of a transposon mutagenesis method . Here, we describe a novel technique for random transposon mutagenesis using a mariner-based transposon into C . jejuni strain 480 . Insertions were random, as demonstrated by Southern blot analysis and insertional junction sequencing . We have demonstrated, for the first time, random in vivo transposon mutagenesis of C . jejuni. J Immunol, 2000 Sep 1, 165(5), 2612 - 8 Expression of the cytolethal distending toxin (Cdt) operon in Actinobacillus actinomycetemcomitans: evidence that the CdtB protein is responsible for G2 arrest of the cell cycle in human T cells; Shenker BJ et al.; We have previously shown that Actinobacillus actinomycetemcomitans produces an immunosuppressive factor that is encoded by the cdtB gene, which is homologous to a family of cytolethal distending toxins (Cdt) expressed by several gram-negative bacteria . In this study, we report that the cdt locus in A . actinomycetemcomitans is composed of five open reading frames, designated orf1, orf2, cdtA, cdtB, and cdtC . The deduced amino acid sequences of the five open reading frames are highly conserved among A . actinomycetemcomitans strains 652, Y4, 29522, and HK1651 . There is also strong homology with the Cdt proteins of Haemophilus ducreyi (87-91%), but only partial homology with that of Campylobacter jejuni and Escherichia coli (29-48%) . Analysis of A . actinomycetemcomitans mRNA by RT-PCR suggests that the two small open reading frames upstream of cdtA are coexpressed with cdtA, cdtB, and cdtC . We next utilized a series of plasmids that express various combinations of the cdt genes to determine their requirement for expression of immunoinhibitory activity . Cell extracts of E . coli transformed with each of the plasmids were tested for their capacity to induce G2 arrest in the cell cycle of PHA-activated human T cells . These experiments suggest that expression of cdtB alone is sufficient to induce G2 arrest in human T cells, but do not exclude the possibility that cdtC also contributes to cell cycle arrest . The implications of our results with respect to the function of the individual Cdt proteins are discussed. J Food Prot, 2000 Aug, 63(8), 1043 - 8 Rapid detection of Salmonella typhimurium in chicken carcass wash water using an immunoelectrochemical method; Che YH et al.; An immunoelectrochemical method coupled with immunomagnetic separation was developed for rapid detection of Salmonella Typhimurium in chicken carcass wash water . Samples of chicken carcass wash water were inoculated with Salmonella Typhimurium at different cell numbers . Possible nonspecified inhibitors in the wash water were minimized by filtration and centrifugation . An approximately 9.4% loss of Salmonella cells was found after filtration (P < 0.01) . The samples were mixed with anti-Salmonella-coated magnetic beads (ASCMB) and alkaline phosphatase-labeled anti-Salmonella (APLAS) to form ASCMB-Salmonella-APLAS conjugates . The conjugates were separated from the solution using a magnetic separator and then incubated with phenylphosphate substrate to produce phenol . The number of Salmonella was determined by measuring the phenol concentration using an amperometric tyrosinase carbon paste electrode in a flow injection analysis system . Under optimized parameters (1 mM MgCl2, 0.2 microg/ml APLAS, and 1 mM phenylphosphate in pH 7.0 Tris buffer solution), Salmonella Typhimurium in chicken carcass wash water could be identified and enumerated within 2.5 h with a detection limit of 5 x 10(3) CFU/ml . A linear relationship on a log-log scale was found between Salmonella cell number and the peak current ratio for Salmonella concentrations ranging from 10(3) to 10(7) CFU/ml (R2 = 0.963) . The peak currents of multibacteria samples, containing Salmonella Typhimurium, Listeria monocytogenes, and Campylobacter jejuni, were not significantly different from Salmonella-only samples (P > 0.01). N Z Med J, 2000 Jul 14, 113(1113), 281 - 4 Economic cost to New Zealand of foodborne infectious disease; Scott WG et al.; AIMS: To estimate the annual economic cost to New Zealand of foodborne infectious disease . METHODS: Annual incidence rates were combined with unit cost data to derive estimates of the annual economic cost to society of each foodborne infectious disease . Market prices and wages were used as proxies for the unit costs of resource utilisations . A decision analytic model was developed to estimate the costs of each disease and to undertake sensitivity analysis . RESULTS: There are an estimated 119 320 episodes of foodborne infectious disease per year in New Zealand (3241 per 100 000 population) . The total cost of these cases was $55.1 million ($462 per case) made up of direct medical costs of $2.1 million, direct non-medical costs of $0.2 million, indirect cost of lost productivity of $48.1 million, and intangible cost of loss of life of $4.7 million . Campylobacteriosis generated most of the costs . Lost productivity was the major cost component for all diseases . The total cost of potentially foodborne infectious disease was estimated to be $88.8 million . Broad estimates of additional costs due to cases of infectious intestinal diseases caused by non-foodborne pathogens or for which no pathogen is identified could raise the cost to $215.7 million . CONCLUSION: The findings imply that resources of $55 million could be devoted to prevention of foodborne infectious disease . Efforts should focus on lowering the incidence of campylobacteriosis as this disease accounts for most of foodborne illness costs. Rev Sci Tech, 2000 Aug, 19(2), 638 - 61 Ostrich diseases; Verwoerd DJ; Scientific knowledge of ostrich diseases is incomplete and very fragmented, with specific details on technical aspects of diagnostic and/or screening tests completely absent in most cases . Salmonella Typhimurium is common in multispecies collections and causes mortality in chicks younger than three months on commercial farms, but is rarely found in chicks older than six months, or slaughter birds of twelve to fourteen months in southern Africa . Campylobacter jejuni and Chlamydia psittaci are occasionally reported, mainly in young ostriches, but both remain a diagnostic challenge . Crimean-Congo haemorrhagic fever is transmitted to domestic animals including ostriches, principally by ticks of the genus Hyalomma . In the ostrich, the disease causes no clinical symptoms during a viraemia of approximately four days . Spongiform encephalopathy has not been reliably reported in ostriches, while anthrax has occurred rarely in modern times but was reportedly an important cause of death approximately 100 years ago in South Africa . Salmonella Gallinarum and S . Pullorum are unknown in ostriches . Pasteurella multocida occurs but is easily contained with antibiotics . Mycoplasma spp . are regularly found in an upper respiratory disease syndrome complicated by opportunistic bacterial pathogens . Ostriches of all ages are susceptible to challenge by velogenic Newcastle disease virus (NDV), but standard inactivated La Sota poultry vaccines can stimulate protective immunity lasting over six months . The viraemic period in vaccinated slaughter ostriches is between nine and eleven days and there are no indications of a carrier state or presence of the virus in the meat or any other tissues after this period, with peak immunoglobulin G response reached on day fourteen post infection . Haemagglutination inhibition tests are significantly less sensitive and less specific than enzyme-linked immunosorbent assays . Cloacal and choanal swabs used for direct virological screening in clinically affected cases (field and experimental) could not detect NDV . All avian influenza isolates reported from ostriches have been non-pathogenic to poultry, even the H5 and H7 subtypes . Some of the latter have been associated with mortality of ostrich chicks in localised outbreaks during periods of inclement weather and with significant wild bird (waterfowl) contact . Borna disease causes a nervous syndrome in ostrich chicks, but to date, has only been reported in Israel . Eastern and Western equine encephalomyelitides cause fatal disease in ostriches and other ratites, with mortality ranging from less than 20% to over 80% in affected flocks . These diseases are present in North, Central and South America where the associated ornithophilic mosquito vectors occur . Equine and human vaccines are apparently safe and efficacious in ratites . Wesselsbron disease, infectious bursal disease (type 2), adenovirus and coronavirus infections have been reported from ostriches but the significance of these diseases is unclear . Due to the paucity of data regarding ostrich diseases and the unvalidated state of most poultry tests in this unique group of birds, strict observation of a pre-slaughter quarantine of thirty days is strongly advised, whilst live exports and fertile eggs should be screened through the additional use of sentinel chickens and/or young ostriches. Rev Sci Tech, 2000 Aug, 19(2), 376 - 95 Campylobacter infection of commercial poultry; Shane SM; Campylobacter jejuni, a widespread food-borne pathogen is responsible for enteritis in the populations of both industrialised and developing nations and is acquired by consumption of contaminated water, milk and food products . Contaminated poultry meat is regarded as an important source of campylobacteriosis, with both commercial broiler and turkey growing flocks infected at two to three weeks of age by direct and indirect horizontal exposure . Non-chlorinated water is regarded as a vehicle of infection, followed by rapid intraflock dissemination . Intensification in the poultry industry has contributed to the increased prevalence rates on carcasses associated with increased stocking density and mechanized processing which are inherent to the high efficiency dictated by a competitive market . Currently, pre- and post-harvest control measures may ameliorate the problem of Campylobacter infection in consumers . Refrigerated storage and transport of red meat and poultry, appropriate handling and food preparation, and thorough cooking reduce the possibility of food-borne infection . In view of the world-wide distribution of C . jejuni infection and the multiplicity of sources, including non-pasteurised milk and contaminated water, it is inappropriate to impose trade restrictions on poultry meat based on the detection of campylobacters. J Antimicrob Chemother, 2000 Aug, 46(2), 303 - 6 Activity of antibiotics used in human medicine for Campylobacter jejuni isolated from farm animals and their environment in Lancashire, UK; Piddock LJ et al.; A retrospective study of 96 Campylobacter jejuni isolated from farm animals and the environment showed that most were less susceptible than the NCTC type strain to nalidixic acid (MICs 4-32 mg/L), ciprofloxacin (MICs 1-2 mg/L) and erythromycin (MICs 16-64 mg/L), but had similar susceptibility to tetracycline (MICs 4-8 mg/L) and kanamycin (MICs 4-8 mg/L) . None had the high MICs of ciprofloxacin (>32 mg/L) or erythromycin (1024 mg/L) typically associated with clinical resistance in this species . Some farms used antimicrobial agents, but there was no obvious association between the use of agents and the susceptibility of the isolates. Mol Microbiol, 2000 Aug, 37(3), 501 - 14 Phase variation of a beta-1,3 galactosyltransferase involved in generation of the ganglioside GM1-like lipo-oligosaccharide of Campylobacter jejuni; Linton D et al.; Ganglioside mimicry by Campylobacter jejuni lipo-oligosaccharide (LOS) is thought to be a critical factor in the triggering of the Guillain-Barre and Miller-Fisher syndrome neuropathies after C . jejuni infection . The combination of a completed genome sequence and a ganglioside GM1-like LOS structure makes C . jejuni NCTC 11168 a useful model strain for the identification and characterization of the genes involved in the biosynthesis of ganglioside-mimicking LOS . Genome analysis identified a putative LOS biosynthetic cluster and, from this, we describe a putative gene (ORF Cj1139c), which we have termed wlaN, with a significant level of similarity to a number of bacterial glycosyltransferases . Mutation of this gene in C . jejuni NCTC 11168 resulted in a LOS molecule of increased electrophoretic mobility, which also failed to bind cholera toxin . Comparison of LOS structural data from wild type and the mutant strain indicated lack of a terminal beta-1,3-linked galactose residue in the latter . The wlaN gene product was demonstrated unambiguously as a beta-1,3 galactosyltransferase responsible for converting GM2-like LOS structures to GM1-like by in vitro expression . We also show that the presence of an intragenic homopolymeric tract renders the expression of a functional wlaN gene product phase variable, resulting in distinct C . jejuni NCTC 11168 cell populations with alternate GM1 or GM2 ganglioside-mimicking LOS structures . The distribution of wlaN among a number of C . jejuni strains with known LOS structure was determined and, for C . jejuni NCTC 12500, similar wlaN gene phase variation was shown to occur, so that this strain has the potential to synthesize a GM1-like LOS structure as well as the ganglioside GM2-like LOS structure proposed in the literature. J Clin Microbiol, 2000 Aug, 38(8), 3076 - 9 Rapid detection of Campylobacter jejuni in stool specimens by an enzyme immunoassay and surveillance for Campylobacter upsaliensis in the greater Salt Lake City area; Hindiyeh M et al.; The Alexon-Trend, Inc . (Ramsey, Minn.), ProSpecT Campylobacter microplate assay was compared with culture on a Campy-CVA plate (Remel, Lenexa, Kans.) and blood-free campylobacter agar with cefoperazone (20 microg/ml), amphotericin B (10 microg/ml), and teicoplanin (4 microg/ml) (CAT medium; Oxoid Limited, Hampshire, England) with 631 patient stool samples . The CAT medium was used to isolate Campylobacter upsaliensis . The enzyme immunoassay (EIA) had a sensitivity and a specificity of 89 and 99%, respectively, and the positive and negative predictive values were 80 and 99%, respectively . Even though we extensively looked for C . upsaliensis in stool samples from patients from the greater Salt Lake City area, we did not isolate this species during the study period . The overall excellent specificity of the EIA allows rapid detection and treatment of positive patients; however, a negative result should be confirmed by culture when clinical suspicion is high. Adolesc Med, 2000 Jun, 11(2), 263 - 78 Infectious diseases of gastrointestinal tract in adolescents; Marsh WW; This article reviews the following gastrointestinal infections: esophagitis, gastritis, duodenitis including duodenal ulcers, and enteritis (gastroenteritis) . The epidemiology, risk factors, microbiology and pathogenesis, diagnosis, treatment, morbidity/mortality, and prevention are discussed in relation to the most important pathogens . The symptoms and pathogenesis of esophagitis caused by Candida albicans and herpes simplex are contrasted with the symptoms of esophagitis caused by Helicobacter pylori and gastroesophageal reflux disease (GERD) . The incidence of gastritis and gastric and duodenal ulcers caused by H . pylori is discussed . The treatment regimens of H . pylori infection recommended by the CDC are presented . Endoscopic findings in esophagitis, gastritis, and duodenal ulcers are presented and discussed . The difference in symptoms caused by viral agents (Norwalk virus), bacterial agents (enterotoxigenic E . coli), and parasites (Giardia lamblia and Cryptosporidium parvum) are compared and contrasted . The symptoms of infections of the terminal small bowel caused by Salmonella and Campylobacter jejuni and the symptoms of pure colonic infection, dysentery, caused by Shigella and enteroinvasive E . coli and Entamoeba histolytica are discussed . The treatment regimens for enteritis are presented. J Infect Dis, 2000 Aug, 182(2), 620 - 3 Epub 2000 Jul 19. Cytolethal distending toxin in avian and human isolates of Helicobacter pullorum; Young VB et al.; Helicobacter pullorum has been isolated from the feces and livers of poultry and is associated with human gastroenteritis . Discrimination of this organism from other enterohepatic Helicobacter species and Campylobacter species has proven difficult . H . pullorum from both avian and human clinical sources has DNA sequence homology and cytotoxic activity that represent a new member of the cytolethal distending toxin (CDT) family of bacterial toxins . CDT is a potential virulence factor in H . pullorum that may serve as a distinguishing phenotype and aid in identification of this organism in veterinary and human clinical samples. J Periodontol, 2000 Jun, 71(6), 885 - 97 "Checkerboard" assessments of periodontal microbiota and serum antibody responses: a case-control study; Papapanou PN et al.; BACKGROUND: We explored the association between subgingival microbial profiles and serum IgG responses to periodontal microbiota in relation to clinical periodontal status . METHODS: One hundred thirty-one (131) periodontitis patients aged 29 to 74 years (mean 51.8) were age- and gender-matched with 74 periodontally intact controls (range 26 to 77, mean 49.3) . Smoking habits and health history were recorded and assessments of plaque, bleeding on probing, probing depth, and attachment level were performed at 6 sites per tooth on all present teeth, excluding third molars . Subgingival plaque samples were obtained from each tooth in one upper and one lower quadrant (maximum 14 samples/subject; 2,440 samples total) and analyzed with respect to 19 species by means of whole genomic DNA probes . Serum IgG antibodies against the same 19 species were assessed by an immunoassay . RESULTS: Cases displayed an average of 22.7 teeth, 20.3 sites with probing depth > or =6 mm, and 18.9 sites with attachment loss > or =6 mm . Corresponding figures for controls were 27.1, 0.1, and 1.0, respectively . Heavy smoking was 3 times more frequent among cases than controls (32.1% versus 9.6%) . Higher levels of Porphyromonas gingivalis, Porphyromonas endodontalis, Prevotella intermedia, Prevotella nigrescens, Prevotella melaninogenica, Bacteroides forsythus, Fusobacterium nucleatum, Treponema denticola, Eubacterium nodatum, Peptostreptococcus micros, and Campylobacter rectus were found in cases and higher levels of Eikenella corrodens, Veillonella parvula, and Actinomyces naeslundii in controls . Cases displayed higher IgG levels against P . gingivalis and Actinobacillus actinomycetemcomitans, while controls displayed higher levels against F . nucleatum, T . denticola, E . nodatum, and Capnocytophaga ochracea . Positive correlations between bacterial colonization and antibody responses were identified for 9 species in controls . In cases, however, statistically significant correlations were observed for only 3 species out of which only one was positive (V . parvula) . Both bacterial levels and antibody responses declined in ages over 55 years . A logistic regression employing selected elements of bacterial colonization and antibody responses as independent variables resulted in 81.1% correct diagnosis, with sensitivity of 83.1%, specificity of 77.8%, positive predictability of 86%, and negative predictability of 73.7% . Smoking did not reach statistical significance in this model . CONCLUSION: A combined microbial colonization/antibody response profile can effectively discriminate between periodontitis patients and periodontally intact controls. FEMS Microbiol Lett, 2000 Aug 1, 189(1), 19 - 24 A variation of the amplified-fragment length polymorphism (AFLP) technique using three restriction endonucleases, and assessment of the enzyme combination BglII-MfeI for AFLP analysis of Salmonella enterica subsp . enterica isolates; Lindstedt BA et al.; We have performed amplified-fragment length polymorphism (AFLP) fingerprinting on a collection of Salmonella enterica subsp . enterica serovar typhimurium strains with a restriction endonuclease combination (BglII and MfeI) that has previously been used successfully for typing Campylobacter jejuni isolates with high resolution . Additionally, a variation of the AFLP assay in which two rare cutting restriction enzymes (XbaI and BsrGI) in combination with the frequent cutter (HinP1I) was examined . The BglII and MfeI enzyme combination offered low resolution for genotyping Salmonella typhimurium isolates and is not recommended for this common serovar . The three-enzyme combination gave a higher discrimination, and is thus a new alternate way of performing AFLP fingerprinting of S . typhimurium. Prev Vet Med, 2000 Aug 10, 46(3), 209 - 23 A longitudinal study of campylobacter infection of broiler flocks in Great Britain; Evans SJ et al.; One hundred flocks associated with five integrated poultry companies were monitored for one production cycle to investigate risk factors for campylobacter infection of poultry broiler flocks . Bacteriological samples were collected from one house of birds on each site at weekly intervals from 3 to 4 weeks of age until the birds were infected with campylobacter or the flock was depopulated (whichever was sooner) . Environmental samples were obtained from 20 houses after cleansing and disinfection of the site before chick arrival . Conventional methods were used for the isolation of campylobacter . Questionnaires were used to collect information on potential risk factors for campylobacter infection . Discrete-time survival analysis was used to assess the influence of various exposures on the age at which the flock was infected with campylobacter.More than 40% of flocks were infected with campylobacter by the time the chicks were 4 weeks old and >90% by 7 weeks . Infection spread rapidly to most birds in a flock . Infection was not predictable by campylobacter status of the last flock reared on the site . (However, because most flocks were infected, the power to detect such an association was poor.) There was no evidence of environmental survival of campylobacters in broiler houses after adequate cleansing and disinfection . The most important predictors of protection from campylobacter were related to effective hygiene barriers (such as housing birds in buildings in a good state of repair, appropriate usage of disinfectant boot dips and a high standard of cleansing and disinfection of the drinking-water equipment) . There was no evidence that rodents were a source of infection (but most sites operated effective vermin-control programmes). Acta Trop, 2000 Jul 21, 76(1), 59 - 63 Zoonotic infections in Nigeria: overview from a medical perspective; Coker AO et al.; Infections of domestic and wild animals that are transmitted directly or by an arthropod vector to humans are a major cause of morbidity and mortality worldwide and particularly in Nigeria . With a population of over 100 million and the need for improved health care delivery, Nigerians are at considerable risk considering the seriousness of these infections . Zoonotic infections that are endemic in Nigeria include tuberculosis, trypanosomiasis, toxoplasmosis, taeniasis, rabies, lassa fever and yellow fever . Zoonotic food-borne infections (caused by Campylobacter, Salmonella and Escherichia coli O157:H7) and cryptosporidiosis are emerging . Sporadic cases such as strongyloidiasis, ascariasis, leptospirosis, scabies, pentastomiasis and African histoplasmosis have been reported . There is a need to determine the prevalence of tick-borne zoonoses . Prevention and control of zoonoses in humans is by vaccination, treatment and health education . As a first measure to improve control, the link between veterinary and medical officers, which is presently very weak, needs to be strengthened . Furthermore, regional multidisciplinary approaches to the control of zoonotic infections should be adopted in West Africa, which take into consideration the huge inter-border traffic. FEMS Microbiol Lett, 2000 Jul 15, 188(2), 115 - 8 The iron-responsive regulator Fur of Campylobacter jejuni is expressed from two separate promoters; van Vliet AH et al.; A lacZ-based reporter gene system was used to identify the promoter of the Campylobacter jejuni iron-responsive gene regulator Fur . In other Gram-negative bacteria, the fur promoter is usually located directly upstream of the fur gene and is often autoregulated in response to iron . In this study we demonstrate that expression of the C . jejuni fur gene is controlled from two promoters located in front of the first and second open reading frames upstream of fur . Neither of these promoters was iron-regulated, and the presence of both promoters in front of fur gives higher expression of the lacZ reporter than with either promoter alone . Expression from two distal promoters might be a mechanism for regulating the level of the C . jejuni Fur protein in response to unknown stimuli. Clin Infect Dis, 2000 Jul, 31(1), 192 - 6 Acute community-acquired diarrhea requiring hospital admission in Swiss children; Essers B et al.; In order to ascertain the prevalence of agents that cause childhood diarrheal illness, stool specimens of 312 consecutive children with community-acquired diarrhea requiring admission were evaluated . Pathogens were detected in 166 (53%) of the 312 children (>/=2 pathogens in 28 children): Rotavirus (n=75), Salmonella spp . (n=37), Campylobacter spp . (n=24), Shigella spp . (n=5), Giardia spp . (n=4), Yersinia spp . (n=2), Aeromonas spp . (n=15), Cryptosporidium (n=15), enteropathogenic Escherichia coli (n=13), enterotoxigenic E . coli (n=7), and enterohemorrhagic E . coli (n=5) . In conclusion, acute childhood diarrheal illness pathogens, such as Aeromonas, Cryptosporidium, and diarrheagenic E . coli, account for a large proportion of patients with a microbiologically positive stool specimen. Br J Biomed Sci, 2000, 57(2), 137 - 41 Deoxyribonucleic acid restriction digest patterns in Campylobacter species: a comparison with Penner serotype; Smith SI et al.; Diversity, based on restriction fragment length polymorphism (RFLP) analysis, was studied in 48 strains of Campylobacter, comprising 27 chicken and 21 human strains of C . jejuni and C . coli, using genomic Southern hybridisation . Restriction digests of chromosomal DNA were prepared by treating with HaeIII and probed using a C . jejuni DNA probe . Nineteen distinct hybridisation patterns were identified, and differences in hybridisation pattern between members of the two species, and in individual strains of the same species, were seen . The method described proved more discriminatory than the Penner serotype, as strains from the same serotype were distinguished . The relative simplicity of the patterns obtained, together with the apparent diversity identified among individual strains and species, suggests that DNA fingerprinting using the C . jejuni DNA probe could be a useful identification method in epidemiological studies of Campylobacter infection in Nigeria. Emerg Infect Dis, 2000 Jul-Aug, 6(4), 412 - 4 Primary care surveillance for acute bloody diarrhea, Wales; Chalmers RM et al.; A sentinel group of primary-care physicians in Wales actively reported cases of acute bloody diarrhea from February 1997 through December 1998 . The estimated annual rate was 18 cases per 100,000 population . Most (80%) cases were due to Campylobacter or Salmonella; however, 18% were undiagnosed. Commun Dis Public Health, 2000 Jun, 3(2), 101 - 5 Survey of local authority approaches to investigating sporadic cases of suspected food poisoning; Rooney R et al.; Local authorities in England were surveyed in 1997 to document their approach to investigating sporadic cases of suspected food poisoning, the data they collect, and their objectives for doing so . Three hundred and nineteen of the 359 authorities (89%) responded . The main objectives stated were to prevent spread of infection, detect outbreaks, educate the public, and identify risk factors for infection . Most outbreaks of food poisoning were detected as a result of complaints from members of the public . Investigation of Salmonella spp and Vero cytotoxin producing Escherichia coli O157 appeared to have a higher priority than campylobacter infection and viral gastroenteritis . Eighty-five per cent (266/311) of local authorities had agreed timescales for investigating cases and 54% (165/306) reported using computer technology to collate and analyse data on sporadic cases of food poisoning . Almost all (243/244) respondents supported the idea of having a new national system for the collation of such data and 79% (244/309) expressed interest in piloting a new standard form for enhanced surveillance of food poisoning. Infect Immun, 2000 Aug, 68(8), 4384 - 90 Involvement of a plasmid in virulence of Campylobacter jejuni 81-176; Bacon DJ et al.; Campylobacter jejuni strain 81-176 contains two, previously undescribed plasmids, each of which is approximately 35 kb in size . Although one of the plasmids, termed pTet, carries a tetO gene, conjugative transfer of tetracycline resistance to another strain of C . jejuni could not be demonstrated . Partial sequence analysis of the second plasmid, pVir, revealed the presence of four open reading frames which encode proteins with significant sequence similarity to Helicobacter pylori proteins, including one encoded by the cag pathogenicity island . All four of these plasmid-encoded proteins show some level of homology to components of type IV secretion systems . Mutation of one of these plasmid genes, comB3, reduced both adherence to and invasion of INT407 cells to approximately one-third that seen with wild-type strain 81-176 . Mutation of comB3 also reduced the natural transformation frequency . A mutation in a second plasmid gene, a virB11 homolog, resulted in a 6-fold reduction in adherence and an 11-fold reduction in invasion compared to the wild type . The isogenic virB11 mutant of strain 81-176 also demonstrated significantly reduced virulence in the ferret diarrheal disease model . The virB11 homolog was detected on plasmids in 6 out of 58 fresh clinical isolates of C . jejuni, suggesting that plasmids are involved in the virulence of a subset of C . jejuni pathogens. Infect Immun, 2000 Aug, 68(8), 4378 - 83 Helicobacter pylori urease suppresses bactericidal activity of peroxynitrite via carbon dioxide production; Kuwahara H et al.; Helicobacter pylori can produce a persistent infection in the human stomach, where chronic and active inflammation, including the infiltration of phagocytes such as neutrophils and monocytes, is induced . H . pylori may have a defense system against the antimicrobial actions of phagocytes . We studied the defense mechanism of H . pylori against host-derived peroxynitrite (ONOO(-)), a bactericidal metabolite of nitric oxide, focusing on the role of H . pylori urease, which produces CO(2) and NH(3) from urea and is known to be an essential factor for colonization . The viability of H . pylori decreased in a time-dependent manner with continuous exposure to 1 microM ONOO(-), i.e., 0.2% of the initial bacteria remained after a 5-min treatment without urea . The bactericidal action of ONOO(-) against H . pylori was significantly attenuated by the addition of 10 mM urea, the substrate for urease, whereas ONOO(-)-induced killing of a urease-deficient mutant of H . pylori or Campylobacter jejuni, another microaerophilic bacterium lacking urease, was not affected by the addition of urea . Such a protective effect of urea was potentiated by supplementation with exogenous urease, and it was almost completely nullified by 10 microM flurofamide, a specific inhibitor of urease . The bactericidal action of ONOO(-) was also suppressed by the addition of 20 mM NaHCO(3) but not by the addition of 20 mM NH(3) . In addition, the nitration of L-tyrosine of H . pylori after treatment with ONOO(-) was significantly reduced by the addition of urea or NaHCO(3), as assessed by high-performance liquid chromatography with electrochemical detection . These results suggest that H . pylori-associated urease functions to produce a potent ONOO(-) scavenger, CO(2)/HCO(3)(-), that defends the bacteria from ONOO(-) cytotoxicity . The protective effect of urease may thus facilitate sustained bacterial colonization in the infected gastric mucosa. Curr Biol, 2000 Jun 29, 10(13), R498 - 501 The controlled chaos of shifty pathogens; Faguy DM; Bacterial pathogens use novel mechanisms to vary their surface structures . Three new genome sequences provide a perspective on these mechanisms in Borrelia burgdorferi, Neisseria meningitidis, and Campylobacter jejuni, which cause lyme disease, meningitis and gastroenteritis, respectively. Eur J Surg Suppl, 1999, (584), 17 - 25 Efficacy and potential clinical applications of Pentaglobin, an IgM-enriched immunoglobulin concentrate suitable for intravenous infusion; Lissner R et al.; OBJECTIVE: Characterisation of the antibodies against important human pathogens in two immunoglobulin preparations: Intraglobin F and IgM-enriched Pentaglobin . DESIGN: In vitro assay of antibody titre using bacterial outer-membrane proteins, lipopolysaccharides (LPS), and exotoxins of clinically relevant bacteria . METHODS: Antibody reactivities measured by ELISA and immunoblot techniques against antigens from bacteria that cause sepsis, antibiotic-resistant nosocomial pathogens, and enteric pathogens . RESULTS: IgG anti-LPS reactivity was present in both study drugs . Specific IgM antibodies against LPS of gram-negative bacteria that cause sepsis were also detected in the IgM-enriched Pentaglobin . IgG-reactivity against gram-positive multiresistant strains of Staphylococcus aureus (S . aureus) were detectable in both preparations . IgG and IgM antibodies present against Yersinia outer proteins and Campylobacter jejuni (C . jejuni) outer membrane proteins were detected in Pentaglobin . Both preparations reacted against alpha toxin of S . aureus and streptolysin of Streptococcus pyogenes . Pentaglobin showed a strong IgM-reactivity against alpha-haemolysin . CONCLUSION: Our data suggest that infusion of well characterised immunoglobulin preparations might be beneficial for patients with severe infections . This is highly relevant in view of the high pathogenicity of bacteria that cause infections in patients in hospital and the continually increasing antibiotic resistance (particularly methicillin-resistant S . aureus). Mol Biol Evol, 2000 Jul, 17(7), 989 - 1000 Enolase from Trypanosoma brucei, from the amitochondriate protist Mastigamoeba balamuthi, and from the chloroplast and cytosol of Euglena gracilis: pieces in the evolutionary puzzle of the eukaryotic glycolytic pathway; Hannaert V et al.; Genomic or cDNA clones for the glycolytic enzyme enolase were isolated from the amitochondriate pelobiont MASTIGAMOEBA: balamuthi, from the kinetoplastid TRYPANOSOMA: brucei, and from the euglenid EUGLENA: gracilis . Clones for the cytosolic enzyme were found in all three organisms, whereas EUGLENA: was found to also express mRNA for a second isoenzyme that possesses a putative N-terminal plastid-targeting peptide and is probably targeted to the chloroplast . Database searching revealed that ARABIDOPSIS: also possesses a second enolase gene that encodes an N-terminal extension and is likely targeted to the chloroplast . A phylogeny of enolase amino acid sequences from 6 archaebacteria, 24 eubacteria, and 32 eukaryotes showed that the MASTIGAMOEBA: enolase tended to branch with its homologs from TRYPANOSOMA: and from the amitochondriate protist Entamoeba histolytica . The compartment-specific isoenzymes in EUGLENA: arose through a gene duplication independent of that which gave rise to the compartment-specific isoenzymes in Arabidopsis, as evidenced by the finding that the EUGLENA: enolases are more similar to the homolog from the eubacterium Treponema pallidum than they are to homologs from any other organism sampled . In marked contrast to all other glycolytic enzymes studied to date, enolases from all eukaryotes surveyed here (except EUGLENA:) are not markedly more similar to eubacterial than to archaebacterial homologs . An intriguing indel shared by enolase from eukaryotes, from the archaebacterium Methanococcus jannaschii, and from the eubacterium Campylobacter jejuni maps to the surface of the three-dimensional structure of the enzyme and appears to have occurred at the same position in parallel in independent lineages. J Agric Food Chem, 2000 Jun, 48(6), 2589 - 94 Kombucha fermentation and its antimicrobial activity; Sreeramulu G et al.; Kombucha was prepared in a tea broth (0.5% w/v) supplemented with sucrose (10% w/v) by using a commercially available starter culture . The pH decreased steadily from 5 to 2.5 during the fermentation while the weight of the "tea fungus" and the OD of the tea broth increased through 4 days of the fermentation and remained fairly constant thereafter . The counts of acetic acid-producing bacteria and yeasts in the broth increased up to 4 days of fermentation and decreased afterward . The antimicrobial activity of Kombucha was investigated against a number of pathogenic microorganisms . Staphylococcus aureus, Shigella sonnei, Escherichia coli, Aeromonas hydrophila, Yersinia enterolitica, Pseudomonas aeruginosa, Enterobacter cloacae, Staphylococcus epidermis, Campylobacter jejuni, Salmonella enteritidis, Salmonella typhimurium, Bacillus cereus, Helicobacterpylori, and Listeria monocytogenes were found to be sensitive to Kombucha . According to the literature on Kombucha, acetic acid is considered to be responsible for the inhibitory effect toward a number of microbes tested, and this is also valid in the present study . However, in this study, Kombucha proved to exert antimicrobial activities against E . coli, Sh . sonnei, Sal . typhimurium, Sal . enteritidis, and Cm . jejuni, even at neutral pH and after thermal denaturation . This finding suggests the presence of antimicrobial compounds other than acetic acid and large proteins in Kombucha. Rev Cubana Med Trop, 1999 Jan-Apr, 51(1), 14 - 9 {Bacterial origin of acute diarrhea in Merida, Venezuela}; Vizcaya Delgado LE et al.; 464 stool specimens from children under 5 with acute diarrheal disease and other 149 specimens from the control group were studied from July, 1993, to May, 1995 . The specimens were collected at the Pediatric Emergency Department of the Autonomous Institute of the Teaching Hospital of Los Andes, Merida, Venezuela . The presence of the internationally recommended bacterial, parasitary and viral agents was investigated . The commonest bacteria isolated as unique pathogens were: Shigella (42.85%), Shigella sonnei, the most found, (66.67%), and the thermotolerant Campylobacter, Aeromonas sp . and enteropathogenous Escherichia coli, with 15; 15 and 13.5%, respectively . 6.5% of parasites and 24.12% of Rotavirus were also found . It was concluded that in the period of time under study the infectious and mainly, the bacterial origin is an important cause of acute diarrheal disease in Merida. Lett Appl Microbiol, 2000 Jul, 31(1), 77 - 81 Development of a direct viable count procedure for the investigation of VBNC state in Listeria monocytogenes; Besnard V et al.; A viable but non-culturable (VBNC) bacterial state was originally detected in studies in environmental microbiology . In particular, this state has been demonstrated for a number of human pathogens (Escherichia coli, Salmonella enteritidis, Vibrio cholerae, Legionella pneumophila and Campylobacter jejuni) . The presence of VBNC cells poses a major public health problem since they cannot be detected by traditional culturing methods and the cells remain potentially pathogenic under favourable conditions . But, as far as we know, the VBNC state has not been yet described in Listeria monocytogenes . In most studies, this has been assessed by the Kogure procedure based on cellular elongation in the presence of DNA gyrase inhibitors . The antibiotic used was nalidixic acid in order to prevent DNA replication, only efficient in Gram-negative bacteria studies . In this study, we describe a new DVC procedure to detect and count viable of L . monocytogenes suspended in filtered, sterilized distilled water . We used different concentrations of ciprofloxacin, efficient both in Gram-negative and Gram-positive bacteria . Bacteria cells were removed and resuspended in BHI broth, with yeast extract and ciprofloxacin . The mixture was incubated at different incubation times at 37 degrees C . After different incubation times, cells were filtered through an isopore polycarbonate black membrane filter and covered with a DAPI solution or orange acridine . The filters were prepared and examined by epifluorescence microscopy . Elongated cells were counted as viable cells, whereas normal size was regarded as nonactive ones . This method allows determination of ciprofloxacin concentration and incubation time optimal to detect maximum viable cells percentage in L . monocytogenes. Lett Appl Microbiol, 2000 Jul, 31(1), 14 - 9 Efficacy of flagellin gene typing for epidemiological studies of Campylobacter jejuni in poultry estimated by comparison with macrorestriction profiling; Petersen L et al.; Thirty isolates of Campylobacter jejuni isolated from 29 different Danish broiler flocks were chosen for the evaluation of PCR-Fla typing as a genotyping tool . Except for two isolates that originated from the same broiler flock, the isolates were clearly distinguishable on basis of their macrorestriction profiles using the restriction endonucleases SmaI and KpnI . PCR-Fla typing of the 30 isolates yielded 16 distinct genotypes, whereas one isolate was untypeable by this method . The dominant PCR-Fla type (1/1) was shared by eight isolates, and five additional Fla groups containing two or three isolates were obtained . The PCR-Fla type of one isolate changed spontaneously after five subcultures, illustrating the relative plasticity of the gene locus . Comparison of MRPs within and between Fla-types support the view that some PCR-Fla types may be conserved within clonal lines . It is concluded that PCR-Fla typing is useful as a genotyping tool in large-scale epidemiological studies but that additional analyses with other methods are required to properly define interstrain relationships. Rev Soc Bras Med Trop, 2000 Mar-Apr, 33(2), 223 - 4 The comparison of the Butzler medium, filtration technique and their association with isolation of Campylobacter ssp; Modolo JR; Campylobacter was isolated in 178 out of 622 stool samples (200 porcine, 220 bovine, and 202 canine) . From these 178 samples, the microorganism was identified in 64 samples (36%) isolated only in Butzler selective medium (BSM), 34 samples (19%) using filtration technique (FT), and in 80 samples (45%) using both BSM and FT . Comparison of the proportion of positivity using both techniques showed a significant value (chi2 = 9,184; p > 0.001); BSM (36%) being more efficient than FT (19%) . The use of both techniques yielded the highest isolation positivity (45%). Appl Environ Microbiol, 2000 Jul, 66(7), 3110 - 2 Roles of Fe superoxide dismutase and catalase in resistance of Campylobacter coli to freeze-thaw stress; Stead D et al.; We demonstrated that oxidative stress plays a role in freeze-thaw-induced killing of Campylobacter coli following analysis of mutants deficient in key antioxidant functions . Superoxide anions, but not H(2)O(2), were formed during the freeze-thaw process . However, a failure to detoxify superoxide anions may lead to spontaneous disproportionation of the radicals to H(2)O(2). Zh Mikrobiol Epidemiol Immunobiol, 2000 Jan-Feb, (1), 95 - 103 {The potentials for the laboratory diagnosis of infections caused by bacteria in the genus Campylobacter}; Vorob'ev AA et al.; Information on the laboratory diagnostics of Campylobacter infections, carried out with the use of traditional method and molecular biology methods which based on the study of the genetic apparatus of infective agents, is summarized . Classical bacteriological analysis ensuring the isolation and biochemical identification of bacteria, up to the determination of their species, complex bacteriological analysis permitting their isolation and phenotypic identification, as well as genotypic diagnostics in reference laboratories (the detection and identification of infective agents by means of molecular hybridization or gene amplification), may be used. Symp Ser Soc Appl Microbiol, 2000, (29), 144S - 148S Food safety through the meat supply chain; Attenborough M et al.; Food poisoning in humans can be caused by many different bacterial genera . While the incidence of food poisoning in England, Wales and Scotland from Salmonella has reached a plateau, there has been an increase in the incidence from Campylobacter . The incidence from Escherichia coli O157:H7 rose to 1997 but declined slightly in 1998 (data from the Public Health Laboratory Service and the Scottish Centre for Infection and Environmental Health) . This organism has a high virulence in humans and a very low infective dose . Infection can produce a wide range of responses, including death . The low infective dose presents a major threat . The organism is relatively heat-sensitive and the cooking of food products to achieve a centre core temperature of 70 degrees C for 2 min is sufficient to destroy it . It is relatively acid-tolerant and will survive for several weeks at pH 4.2 . Several foodstuffs, as well as water, have been implicated in world-wide outbreaks . The E . coli O157:H7 food-borne outbreak in Lanarkshire in 1996 led to 21 fatalities . The Pennington Group report, issued in April 1997, reported on the circumstances leading to this outbreak, the implications for food safety and the lessons to be learnt . Four areas covered within the Pennington Group report specific to meat hygiene are reviewed in this paper . On-farm practices must ensure the presentation of clean animals for slaughter . There is a requirement for the development and introduction of risk assessment techniques based upon Hazard Analysis of Critical Control Points in abattoirs, and the Meat and Livestock Commission (MLC) is producing a manual for use by the abattoir sector . The Pennington report stated that there was a need for research into the potential use of end-process treatments such as steam pasteurization . The MLC is involved in evaluating such a system . Meat production premises and butchers' shops in England are introducing HACCP through an MLC scheme funded by the Department of Health . At the point of consumption, food safety is improved by the provision of practical guidelines regarding the handling of meat and meat products . These are distributed at retail outlets and communicated to secondary schools via MLC's educational publications. Zh Mikrobiol Epidemiol Immunobiol, 1999 Nov-Dec, (6), 27 - 30 {A nutrient medium for the isolation and cultivation of Campylobacter}; Temirkhanova ZU et al.; Campylobacter agar, nutrient medium intended for the isolation of bacteria of the genus Campylobacter from clinical material, has been developed . The composition of the medium includes sprat hydrolysate, aerotolerant additive (ferric sulfate--oxide, sodium pyruvate, sodium pyrosulfite), sodium glutaminate, agar . The selective properties of the medium are ensured by introducing the mixture of antibiotics consisting of polymyxin B, rifampicin, amphotericin B, ristomycin . The balanced composition of Campylobacter agar ensures the aerotolerance of Campylobacter organisms and gives the optimal conditions for their growth when the inoculated material is cultivated in the atmosphere made up of the mixture of three gases (5% of oxygen, 10% of carbon dioxide, 85% of nitrogen), as well as under the conditions of a "candle vessel" . The medium suppresses the development of the associative microflora diluted 10(-1) . As shown in the trial of the quality of Campylobacter agar by the inoculation of material taken from patients with acute enteric infections, agricultural animals and monkeys, the medium has pronounced selective, properties with regard to extraneous microflora, while ensuring the isolation of Campylobacter on the level of the control medium. J Periodontol, 2000 May, 71(5), 816 - 24 Features of severe periodontal disease in a teenager with Chédiak-Higashi syndrome; Delcourt-Debruyne EM et al.; BACKGROUND: Chediak-Higashi syndrome (C-HS) is a rare congenital disease characterized by defective neutrophil function with abnormal lysosomal inclusions, neutropenia, and reduced chemotaxis . The complete syndrome includes oculocutaneous albinism with photophobia, neurologic features, recurrent infections, and enterocolitis . METHODS: A 14-year-old male C-HS patient was referred to us because of serious periodontal destruction with acute inflamed gingiva and ulcers . Clinical and biological investigations were performed, leading to the diagnosis of C-HS . RESULTS: Laboratory findings included neutropenia and hypergammaglobulinemia . Peripheral blood smears showed giant granules in neutrophils, eosinophils, and granulocytes . Bone marrow smears showed giant inclusions in leukocyte precursor cells . These granules and inclusions were characteristic of Chediak-Higashi syndrome . Oral radiographic status showed extensive loss of alveolar bone leading, in most cases, to tooth exfoliation . Bacteria often associated with periodontitis were detected in subgingival plaque samples, including Fusobacterium nucleatum, Campylobacter rectus, Prevotella melaninogenica, Peptostreptococcus anaerobius, and Clostridium sp . Biopsies of periodontal tissues for light and electronic microscopic examinations revealed massive bacterial invasion of the epithelial tissue, epithelial cells, and connective tissue . Ultrastructural observations of periodontal polymorphonuclear leukocytes showed defective granulation, with abnormal granules not discharging their lysosomal content against engulfed bacteria . Viable dividing bacteria were found in the cytoplasm . CONCLUSIONS: In this case, early-onset periodontitis seems to be the expression of C-HS granulocyte deficiency . Periodontal treatment of these patients is often unsuccessful . This case report illustrates the importance of the dentist in initiating clinical and biological investigations in such early aggressive periodontitis in young patients. Small Rumin, Res. . 2000 Aug 1, 37(3), 215 - 221 Bioserogroups of Campylobacter species isolated from sheep in Kaduna State, Nigeria; Raji MA et al.; Sheep Campylobacter isolates from Kaduna State were characterized into their species and bioserogrouped . A total of 1100 samples were collected from Kaduna abattoir and National Animal Production Research Institute (NAPRI), Shika . The samples were from 250 gallbladder, 250 intestinal contents, 100 fetal stomach contents all from Kaduna abattoir while 250 rectal swabs and 250 vaginal swabs were from the NAPRI Small Ruminant Programme . Of a total of 1100 samples, 39 (3.54%) yielded Campylobacter organisms . The highest isolation rate (6.8%) was from samples of intestinal contents followed by those from gall bladders (4.0%) . Samples from the vaginal and fetuses had the lowest isolation rates (2.80%) and (0%), respectively . Of the 39 Campylobacter isolates from all the sources, (79%) were characterized as C . fetus subsp jejuni, C . coli (13%) and C . laridis (8.0%) . while C . coli and C . laridis were isolated from gall bladder and intestinal contents only . Campylobacter fetus subsp jejuni biotype 1 accounted for 40.3% of the total isolates . C . laridis biotypes I and II were also isolated and accounted for 5% and 3% of the isolates, respectively . 5% of the isolates were not typeable . The serogroups 4 (13%), 36 (10%), 9 (10%), 84 (8%), 29 (5%) and 20 (8%) were the commonest serogroups identified in sheep at two locations surveyed . The isolation of Campylobacter organisms from rectum, vagina, gallbladder, and intestinal contents is a clear indication that sheep serves as a reservoir of this organisms in Nigeria . Similarities between documented human Campylobacter isolates in Nigeria and those in the present study raised the possibility of cross-transmission between sheep and man . It is concluded that biotyping and serotyping can be used for epidemiological study of campylobacteriosis due to Campylobacter jejuni in sheep in Kaduna State of Nigeria. FEMS Immunol Med Microbiol, 2000 Jul, 28(3), 257 - 63 Bacterial, viral and parasitic enteric pathogens associated with acute diarrhea in hospitalized children from northern Jordan; Youssef M et al.; To determine the etiology of acute diarrhea in Jordanian children under 5 years of age, we examined stool samples from 265 children admitted to the pediatric ward at Princess Rahma Hospital for Children, Irbid, Jordan, for parasites, rotavirus and enteric bacteria . Using both traditional and molecular diagnostic techniques, we detected enteropathogens in 66.4% of patients with diarrhea . A single enteric pathogen was detected in 50.9% of the children, and multiple pathogens were detected in 15.5% . The prevalence of enteropathogens identified was as follows: rotavirus (32.5%), enteropathogenic Escherichia coli (12.8%), enteroaggregative E . coli (10.2), enterotoxigenic E . coli (5.7%), Shigella spp . (4.9%), Entamoeba histolytica (4.9%), Salmonella spp . (4.5%), Campylobacter jejuni/coli (1.5%), Cryptosporidium spp . (1.5%), enteroinvasive E . coli (1.5%), eae-, Ehly-positive E . coli (0.8%), Giardia lamblia (0 . 8%) and Yersinia enterocolitica (0.4%) . No Vibrio cholerae, Shiga toxin-producing E . coli, microsporidia, adenovirus or small round virus were detected . Findings from this study demonstrate that rotavirus and several types of diarrheagenic E . coli, which are not screened for during routine examinations of stool samples in public health laboratories, were the most frequently detected enteropathogens in these children . Our findings highlight the value of using a combination of traditional and molecular techniques in the diagnosis of diarrheal disease in this population. Rev Prat, 2000 Apr 1, 50(7), 724 - 30 {Acquired demyelinating neuropathies}; Hahn AF; The acquired demyelinating polyneuropathies with either acute or chronic clinical presentation are considered autoimmune disorders . The Guillain-Barre syndrome is viewed as an acutely reactive and self-limited autoimmune disease, triggered by preceding bacterial or viral infections . There is a particularly strong association with the gastroenteric pathogen, Campylobacter jejuni, and with Cytomegalovirus and Epstein-Barr virus . It is likely that immune response directed towards the infecting organisms are involved in the pathogenesis of Guillain-Barre syndrome by cross-reaction with neural tissues . In the susceptible individual, the infecting organism induces humoral and cellular immune responses that, because of the sharing of homologous epitopes (molecular mimicry), cross-react with ganglioside surface components of peripheral nerve . Immune reactions against target epitopes in the Schwann cell surface membrane or myelin result in acute inflammatory demyelinating neuropathy (90% of cases); reactions against epitopes contained in the axonal membrane cause the acute axonal forms of Guillain-Barre syndrome . Immunomodulation with infusions of IgG or plasma exchange treatments effectively foreshorten the disease course . The immunopathogenesis of the chronic disease forms, chronic inflammatory demyelinating peripheral neuropathy and multifocal motor neuropathy are less well-known . Immunomodulatory treatments with corticosteroid or cytotoxic drug treatments, infusion of Ig or therapeutic plasma exchanges are variably effective . The article outlines the principles and practices of an individualized approach to therapy. J Food Prot, 2000 Jun, 63(6), 807 - 9 Incidence of foodborne illnesses reported by the foodborne diseases active surveillance network (FoodNet)-1997 . FoodNet Working Group; Wallace DJ et al.; In 1997, the Foodborne Diseases Active Surveillance Program (FoodNet) conducted active surveillance for culture-confirmed cases of Campylobacter, Escherichia coli O157, Listeria, Salmonella, Shigella, Vibrio, Yersinia, Cyclospora, and Cryptosporidium in five Emerging Infections Program sites . FoodNet is a collaborative effort of the Centers for Disease Control and Prevention's National Center for Infectious Diseases, the United States Department of Agriculture's Food Safety and Inspection Service, the Food and Drug Administration's Center for Food Safety and Applied Nutrition, and state health departments in California, Connecticut, Georgia, Minnesota, and Oregon . The population under active surveillance for foodborne infections was approximately 16.1 million persons or roughly 6% of the United States Population . Through weekly or monthly contact with all clinical laboratories in these sites, 8,576 total isolations were recorded: 2,205 cases of salmonellosis, 1,273 cases of shigellosis, 468 cases of cryptosporidiosis, 340 of E . coli O157:H7 infections, 139 of yersiniosis, 77 of listeriosis, 51 of Vibrio infections, and 49 of cyclosporiasis . Results from 1997 demonstrate that while there are regional and seasonal differences in reported incidence rates of certain bacterial and parasitic diseases, and that some pathogens showed a change in incidence from 1996, the overall incidence of illness caused by pathogens under surveillance was stable . More data over more years are needed to assess if observed variations in incidence reflect yearly fluctuations or true changes in the burden of foodborne illness. J Vet Med Sci, 2000 May, 62(5), 557 - 60 Risk factors in causing outbreaks of food-borne illness originating in schoollunch facilities in Japan; Michino H et al.; We reviewed records of all outbreaks of food-borne illnesses due to schoollunch in Japan from 1987 through 1996 to determine the risk factors causing these outbreaks . Major ha