Infect Immun, 1994 Sep, 62(9), 3901 - 6 Differential flagellin expression in a flaA flaB+ mutant of Campylobacter jejuni; Wassenaar TM et al.; Campylobacter jejuni 81116 has two genes coding for flagellin, flaA and flaB . Fully motile wild-type C . jejuni bacteria express the flaA gene, with no flaB message being detected . A nonmotile flaA flaB+ mutant, R1, produced detectable levels of flagellin B which was incorporated into truncated flagella . After R1 had invaded INT-407 cells, a variant with increased motility, R1-V2, was isolated . R1-V2 produced full-length flagella and an increased amount of flagellin B . Transcriptional analysis showed that R1-V2 contained more flaB mRNA than its parental strain, R1 . The flaB gene promoter sequence and primer extension experiments confirmed that transcription of the flaB gene is initiated from a sigma 54 promoter . Neither the promoter sequence nor the coding sequence of flaB had changed in R1-V2 . In contrast to R1, R1-V2 no longer produced (truncated) flaA mRNA . The sigma 28 flaA promoter sequence was not changed in R1-V2 . We propose that expression of the two flagellin genes in C . jejuni 81116 is regulated at the transcriptional level, in such a way that predominantly one gene at a time is transcribed . We compared the levels of invasiveness of the wild-type strain, R1, and R1-V2 for INT-407 cells . The shift in expression from flaA to flaB occurred not only during invasion assays but also under different conditions in the absence of eukaryotic cells. Infect Immun, 1994 Sep, 62(9), 3773 - 9 Cell association and invasion of Caco-2 cells by Campylobacter jejuni; Russell RG et al.; Adherence and invasion studies were conducted in monolayers of Caco-2 cells . Three-day-old monolayers were inoculated with Campylobacter jejuni 81-176 at a bacterium/cell ratio of 1,000:1 . Saturation studies demonstrated time- and dose-dependent saturation curves for C . jejuni cell association and invasion into Caco-2 cells . Electron microscopy revealed intracellular C . jejuni located within membrane-bound vacuoles . Cell association and invasion were inhibited by 0.3 and 0.5 M concentrations of various sugars, including D-glucose, D-mannose, and D-fucose . However, there was no inhibition with the corresponding L-sugars, indicating physiological specificity . The inhibition of cell association with phloridzin was less pronounced . There was no inhibition of bacterial entry with monodansylcadaverine or g-strophanthin, indicating that it was unlikely that coated-pit formation is important in the invasion of C . jejuni into Caco-2 cells . Furthermore, there was no inhibition with cytochalasin D, vincristine, or vinblastine . Inhibition of cell association was demonstrated at 4 degrees C . Significantly decreased cell association and invasion were seen in potassium-depleted cells . Treatment of cells with bromelain also caused reduction in the number of C . jejuni binding to cells . A nonmotile aflagellate variant of C . jejuni also showed reduced invasion . The results of this study are consistent with energy-dependent invasion mechanisms . The results do not support an endocytic method of invasion for C . jejuni into Caco-2 cells. J Appl Bacteriol, 1994 Sep, 77(3), 303 - 7 Production and viability of coccoid forms of Campylobacter jejuni; Boucher SN et al.; Studies were conducted into the formation and physiological state of coccoid cells of a strain of the human and animal pathogen Campylobacter jejuni . It was found that growth phase and the presence of chloramphenicol did not affect the rate of shape transformation from spiral to coccoid, while nutrient limitation, aeration of the medium and the presence of free-radical scavengers had profound effects . Coccoid cells were found to reduce the tetrazolium salts INT (2-(p-iodophenyl)-3-(p-nitrophenyl)-5-phenyl tetrazolium chloride) and CTC (5-cyano-2,3-ditolyl tetrazolium chloride) to their respective formazans and this was linked to cellular respiration . However, respiring coccoid cells could not sustain their existence in prolonged adverse conditions, and it was concluded that they represent a degenerative stage rather than a dormant state of the organism. Yonsei Med J, 1994 Sep, 35(3), 314 - 9 Antimicrobial susceptibility of Campylobacter fetus subsp . fetus isolated from blood and synovial fluid; Kwon SY et al.; Campylobacter fetus subsp . fetus is a rare human pathogen, but can cause serious extraintestinal infections . Effective antimicrobial agent is required for the therapy, but we have very limited knowledge on the susceptibility of the organism . In this study, the susceptibility of 25 isolates of the organism to 14 antimicrobial agents was tested by an agar dilution method . Antimicrobial agents with low MIC ranges, in micrograms/ml, were: meropenem < or = 0.25, dirithromycin < or = 0.5, gentamicin < or = 1, amikacin, ofloxacin, tetracycline and erythromycin < or = 2 . The MIC range of cefepime was 0.5-8 micrograms/ml, but those of other beta-lactams were relatively high . All of the isolates were interpreted to be susceptible to cefepime, meropenem, amikacin, gentamicin, ofloxacin, tetracycline and dirithromycin . A significant proportion of the isolates were either intermediate or resistant to ampicillin, cephalothin, cefotaxime, aztreonam, loracarbef and erythromycin . In conclusion, the organism remains susceptible to aminoglycosides and tetracycline . Greater in vitro activity of meropenem, ofloxacin and dirithromycin require clinical evaluation. Diagn Microbiol Infect Dis, 1994 Sep, 20(1), 45 - 7 DU-6859a, a new fluoroquinolone agent . Comparative in vitro activity against enteric pathogens and multiresistant outpatient Escherichia coli; Tomayko JF et al.; The activity of DU-6859a, a new fluoroquinolone antimicrobial agent, was compared with that of ciprofloxacin by agar dilution susceptibility testing against enteric pathogens and multiresistant Escherichia coli . The results indicate that DU-6859a inhibits most of these organisms at concentrations similar to those of ciprofloxacin . DU-6859a showed increased activity compared to ciprofloxacin against Campylobacter species isolates. Mikrobiol Z, 1994 Sep-Oct, 56(5), 51 - 61 {The epidemiological characteristics of the immunological response in campylobacteriosis}; Kirik DL; An analytical survey of literature on the problem of immunological aspects of campylobacteriosis is presented . Antigenic nature of various components of a bacterial cell of these agents is described . Schemes of serotyping of campylobacteria on the basis of thermolabile and thermostable antigens are analyzed . It is shown expedient to use serotyping for determination of an infection source in the epidemiological practice . Proceeding from the published data presented, a conclusion is made that it is necessary to develop the home schemes of serotyping . At the same time a problem on development of new proximate methods promoting immunological indication of campylobacteriosis antigens in various ecological niches is not less urgent and is to be taken into account when planning antiepidemiological measures against the campylobacteriosis infection. J Clin Microbiol, 1994 Sep, 32(9), 2182 - 4 Rapid detection of vancomycin-resistant enterococci; Edberg SC et al.; Campylobacter blood agar with clindamycin incubated in 6% CO2 served as a medium to both screen for vancomycin resistance and select for presumptive enterococci . Colonies that grew on the medium were specifically identified as enterococci within 30 min by the pyroglutamyl-beta-naphthylamide and rapid bile esculin tests . The combination of a selective medium plus rapid enzyme substrate tests offered an inexpensive means to enumerate vancomycin-resistant enterococci from specimens by using readily available reagents. Antimicrob Agents Chemother, 1994 Sep, 38(9), 1879 - 82 Evolution of susceptibilities of Campylobacter spp . to quinolones and macrolides; Sanchez R et al.; Erythromycin, new macrolides, and quinolones are alternatives for the treatment of Campylobacter infections . Concerns related to the emergence of resistance to both groups of drugs have been raised . We studied the evolution of antimicrobial susceptibilities of 275 clinical isolates of microorganisms of the genus Campylobacter isolated in our institution during a 5-year period (1988 to 1992) . The microorganisms studied were C . jejuni (n = 230), C . coli (n = 42), and C . fetus (n = 3) . The overall resistance rates (determined by the agar dilution method and the recommendations of the National Committee for Clinical Laboratory Standards) were as follows: erythromycin, 2.3%; clarithromycin, 2.3%; azithromycin, 1.9%; ciprofloxacin, 28.5%; norfloxacin, 31%; ofloxacin, 26.3%; and nalidixic acid, 36.8% . The evolution of resistance (percent resistance in 1988 versus percent resistance in 1992) was as follows: erythromycin, 2.6 versus 3.1; clarithromycin, 2.6 versus 3.1; azithromycin, 2.6 versus 3.1; ciprofloxacin, 0 versus 49.5; norfloxacin, 2.6 versus 55.5; ofloxacin, 0 versus 45.6; nalidixic acid, 2.6 versus 56.8 . Our data show stable macrolide activity against Campylobacter spp . and the rapid development of quinolone resistance over the last 5 years. J Infect, 1994 Sep, 29(2), 183 - 7 Discrimination by means of pulsed-field gel electrophoresis between strains of Campylobacter jejuni Lior type 4 derived from sporadic cases and from outbreaks of infection; Suzuki Y et al.; The cleavage patterns of the genomic DNA in 42 strains of Campylobacter jejuni Lior type 4, obtained from sporadic cases and from outbreaks of infection were analysed by means of pulsed-field gel electrophoresis (PFGE) . The cleavage of DNA with SmaI and SalI restriction enzymes showed 16 distinct fragment patterns in 20 sporadic isolates, indicating that they were heterogeneous . On the other hand, the patterns of 22 isolates derived from two outbreaks showed the same unique restriction patterns, respectively . PFGE may therefore prove useful for subclassifying strains of C . jejuni Lior type 4 and for discriminating between strains derived from sporadic cases and those derived from outbreaks of infection. Southeast Asian J Trop Med Public Health, 1994 Sep, 25(3), 443 - 8 Exotoxin profiles of Campylobacters isolated in Malaysia; Tee TS et al.; Approximately 57% of clinical and 33% of poultry isolates examined produced a cytotoxin . Cytotoxic activity was detected in 25 (50%) isolates of Campylobacter of which 12 were isolated from bloody diarrhea and 9 from watery stools . The cytotoxin titers were low, ranging from 2 to 16 . The crude filtrates from 50 Campylobacter isolates showed no cytotoxic effect in Vero cells, no fluid accumulation in suckling mice and no hemolytic activity. Antibiot Khimioter, 1994 Sep-Oct, 39(9-10), 36 - 9 {Beta-lactamases of Campylobacter spp . and their use as epidemiologic markers}; Porin AA et al.; The tests of 64 clinical strains of Campylobacter jejuni and 69 strains isolated from hens revealed beta-lactamases in 96.99 per cent of the cultures . The detected enzymes more frequently splitted oxacillin and ampicillin among 5 betalactams used as the substrates (ampicillin, azlocillin, oxacillin, cefoperazon and cefotaxime) . On the whole 24 spectra of the beta-lactamase activity with the evident domination of the AmAzOxCfpCtx spectrum (29.69 per cent) were detected in both the clinical strains and the strains isolated from the hens . A marked correlation of the beta-lactamase spectra of the strains circulating in the humans and hens was observed . Two strains of C . jejuni isolated from the persons of the same family were studied and correlation of their beta-lactamase spectra was noted . The expediency of the determination of the beta-lactamase spectra for the interspecies typing of Campylobacter spp . was suggested. J Clin Microbiol, 1994 Sep, 32(9), 2305 - 6 Confirmation of human Campylobacter concisus isolates misidentified as Campylobacter mucosalis and suggestions for improved differentiation between the two species; On SL; A strain from human diarrhea originally identified as Campylobacter mucosalis (NCTC 12408) was examined by using 64 phenotypic characters . The similarity of this strain to 297 isolates of Campylobacter, Helicobacter, Arcobacter, and related taxa was then determined with a computer-supported data analysis program, MVSP . NCTC 12408 showed closest similarity to 20 type, reference, and field isolates of Campylobacter concisus . These strains were clearly separated from those of C . mucosalis in the numerical analysis of phenotypic tests; a table was constructed from the data used to aid in differentiating these two species in the clinical laboratory . The identity of NCTC 12408 was confirmed as C . concisus by visual comparison of its sodium dodecyl sulfate-polyacrylamide gel whole-cell protein electrophoregram with those of type strains of C . concisus and C . mucosalis . These data suggest that genuine human infection with C . mucosalis has not yet been reported. Gene, 1994 Aug 19, 146(1), 83 - 6 Cloning and expression of the hup encoding a histone-like protein of Campylobacter jejuni; Konkel ME et al.; A Campylobacter jejuni gene, designated hup, that appears to encode a homolog of the histone-like DNA-binding protein, HU, has been cloned, sequenced and expressed in Escherichia coli . Immunoblotting and in vitro transcription/translation analyses revealed a 11-kDa protein that was produced by recombinant plasmids containing hup . The gene contains an open reading frame (ORF) sufficient to encode a protein of 98 amino acids (aa) with a calculated molecular mass of 10,267 Da and a predicted isoelectric point of 10.1 . The deduced aa sequence of the protein, designated HCj, exhibits considerable sequence identity with members of the HU family of proteins from other eubacterial species . The transcription start point was identified by primer extension analysis and appropriately spaced promoter sequences were found which exhibit considerable similarity to E . coli and Bacillus promoters . Southern hybridization analyses indicate that C . jejuni has a single copy of hup. Gene, 1994 Aug 19, 146(1), 31 - 8 Genetic organization of the region upstream from the Campylobacter jejuni flagellar gene flhA; Miller S et al.; Campylobacter jejuni (Cj) is a Gram-bacterium that causes a diarrheal disease in humans . A Cj homolog of the LcrD/FlhA family of proteins was recently described {Miller et al., Infect . Immun . 61 (1993) 2930-2936} . This family includes proteins that are involved in flagellar biogenesis, such as the Cj FlhA protein, but also includes proteins found in invasive pathogens, such as the Yersinia pestis LcrD protein, that play a role in the regulation and/or secretion of virulence-related proteins . Hybridization studies indicated that both the flhA gene and upstream DNA are present in several bacterial species closely related to Cj, including C . fetus, C . lari, C . upsaliensis and C . hyointestinalis . The presence of a second flhA/lcrD homolog was not detected in Cj, indicating that a a separate homolog involved in secretion of virulence proteins may not be present . The 4-kb region immediately upstream from Cj flhA was analyzed . Three open reading frames (ORFs) were found: a 408-nucleotide (nt) gene encoding a homolog of proteins present in Escherichia coli and Desulfovibrio vulgaris, but of unknown function, a 266-nt rpsO gene and a 2823-nt gene encoding a homolog of the Bacillus subtilis SpoIIIE protein . The Cj SpoIIIE homolog had 53% similar or identical amino acids when compared to the B . subtilis protein, and like the B . subtilis protein contained a nt-binding domain and potential transmembrane (TM) regions . All three ORFs were expressed in E . coli minicells, apparently from their own promoters.(ABSTRACT TRUNCATED AT 250 WORDS) Prof Care Mother Child, 1994 Aug-Sep, 4(6), 188 - 9 Health promotion in the home: 1 . Food poisoning: the link with home hygiene; Perry B; The incidence of food poisoning in the UK is increasing substantially, with about 50% arising in the home . Many different organisms cause food-borne infections, including Salmonella, Campylobacter and Shigella . Storing food correctly is important . Many households shop only once a week and food is often kept for several days before being eaten . Standards of domestic kitchen hygiene must be improved if the incidence of food poisoning is to be reduced Hand washing, kitchen cleanliness and disinfection are essential. Bull Tokyo Dent Coll, 1994 Aug, 35(3), 107 - 19 Bacteriological diagnosis of periodontal disease; Okuda K; Dental plaque bacteria cause virtually all the forms of inflammatory periodontal disease . Periodontitis is caused by the specific periodontopathic bacteria, which induce destruction of connective tissue attachment and adjacent alveolar bone . Examinations to identify the infections by Porphyromonas gingivalis, Actinobacillus actinomycetemcomitans, Prevotella intermedia, Campylobacter rectus, Bacteroides forsythus, Fusobacterium nucleatum, Eikenella corrodens and Treponema denticola have recently become essential in diagnosis of periodontal disease . Bacterial examination permits (1) identification of the causative bacteria, (2) assessment of disease activity and (3) monitoring of the effective of periodontal treatments . The author describes details of accurate and rapid methods for detecting periodontopathogens . Transmission of periodontopathic bacteria as clarified by bacterial examination is also discussed in this review . The pathogenic potential of specific bacteria varies among patients and periodontally healthy individuals and can be controlled by host defense mechanisms such as immune responses . The roles of immune responses against periodontopathic bacteria in balance shifts of periodontal disease processes are therefore also discussed in this review. J Dent Res, 1994 Aug, 73(8), 1421 - 8 Interdental supragingival plaque--a natural habitat of Actinobacillus actinomycetemcomitans, Bacteroides forsythus, Campylobacter rectus, and Prevotella nigrescens; Gmur R et al.; The aim of this study was to test the hypothesis that suspected periodontal pathogens form a minor component of the supragingival plaque of individuals without periodontal diseases . Twenty-one dental hygienist trainees with a mean age of 23.5 years were twice sampled for interdental plaque between 1st and 2nd molars in all quadrants . The samples were assessed for Actinobacillus actinomycetemcomitans, Bacteroides forsythus, Campylobacter rectus, Porphyromonas gingivalis, and the Prevotella intermedia/Prevotella nigrescens group of organisms . Bacteria of this group were predominantly P . nigrescens and showed both the highest prevalence (100%) and the highest colonization density of the investigated species . Seven of 21 samples harbored A . actinomycetemcomitans . Serotypes a, b, and c were found in three samples each, while serotype e was present in one sample . Three subjects had two different A . actinomycetemcomitans serotypes . Bacteroides forsythus and C . rectus were detected in 10 (48%) and nine (43%) subjects, respectively . The detected cell numbers accounted for approximately 0.01% to 1% of the sampled flora . In contrast, P . gingivalis was found only in a single sample, which in addition harbored B . forsythus, C . rectus, A . actinomycetemcomitans (serotypes b and e), and P . intermedia . These results suggest that the investigated periodontal bacteria are not "exogenous pathogens", but amphibiotic, opportunistic microorganisms which may have a natural habitat in the supragingival plaque of the interproximal area of molars. J Bacteriol, 1994 Aug, 176(16), 5151 - 5 Nucleotide sequence of the Wolinella succinogenes flagellin, which contains in the antigenic domain two conserved regions also present in Campylobacter spp . and Helicobacter pylori; Schuster SC et al.; Wolinella succinogenes possesses one polar flagellum, which shows a characteristic surface pattern of parallel lines along the axis of the filament in electron microscopic images . We determined the gene sequence of the Wolinella flagellin, which is, as in most other bacteria, the only structural component of the filament . Sequence comparison with other members of the Proteobacteria revealed two highly conserved regions in the central part of the flagellin molecule among Campylobacter spp . and Helicobacter pylori, an area that had previously been described as highly variable . Similar surface patterns are found in related polarly flagellated bacteria, but not in Escherichia coli and Bacillus subtilis, which also lack these conserved regions. J Bacteriol, 1994 Aug, 176(15), 4597 - 609 Phylogenetic and molecular characterization of a 23S rRNA gene positions the genus Campylobacter in the epsilon subdivision of the Proteobacteria and shows that the presence of transcribed spacers is common in Campylobacter spp; Trust TJ et al.; The nucleotide sequence of a 23S rRNA gene of Campylobacter coli VC167 was determined . The primary sequence of the C . coli 23S rRNA was deduced, and a secondary-structure model was constructed . Comparison with Escherichia coli 23S rRNA showed a major difference in the C . coli rRNA at approximately position 1170 (E . coli numbering) in the form of an extra sequence block approximately 147 bp long . PCR analysis of 31 other strains of C . coli and C . jejuni showed that 69% carried a transcribed spacer of either ca . 147 or ca . 37 bp . Comparison of all sequenced Campylobacter transcribed spacers showed that the Campylobacter inserts were related in sequence and percent G+C content . All Campylobacter strains carrying transcribed spacers in their 23S rRNA genes produced fragmented 23S rRNAs . Other strains which produced unfragmented 23S rRNAs did not appear to carry transcribed spacers at this position in their 23S rRNA genes . At the 1850 region (E . coli numbering), Campylobacter 23S rRNA displayed a base pairing signature most like that of the beta and gamma subdivisions of the class Proteobacteria, but in the 270 region, Campylobacter 23S rRNA displayed a helix signature which distinguished it from the alpha, beta, and gamma subdivisions . Phylogenetic analysis comparing C . coli VC167 23S rRNA and a C . jejuni TGH9011 (ATCC 43431) 23S rRNA with 53 other completely sequenced (eu)bacterial 23S rRNAs showed that the two campylobacters form a sister group to the alpha, beta, and gamma proteobacterial 23S rRNAs, a positioning consistent with the idea that the genus Campylobacter belongs to the epsilon subdivision of the class Proteobacteria. J Infect Dis, 1994 Aug, 170(2), 479 - 83 Selection of a gyrA mutant of Mycobacterium tuberculosis resistant to fluoroquinolones during treatment with ofloxacin; Cambau E et al.; A strain of Mycobacterium tuberculosis resistant to ofloxacin was selected in a patient with a long history of multidrug-resistant tuberculosis eventually treated by ofloxacin combined with other second-line drugs . A mutation in the gyrA gene was hypothesized to be the mechanism of acquired resistance to ofloxacin in this strain . Chromosomal DNA of strains MTB1, isolated before treatment and susceptible to ofloxacin (MIC, 1 microgram/mL), and MTB2, isolated during treatment and resistant to ofloxacin (MIC, 32 micrograms/mL), was amplified by polymerase chain reaction (PCR) using two oligonucleotide primers highly homologous to DNA sequences flanking the quinolone resistance-determining region in gyrA of mycobacteria . Comparison of the nucleotide sequences of the 150-bp fragments obtained by PCR revealed a point mutation in MTB2 leading to the substitution of histidine for aspartic acid at a position corresponding to residues involved in quinolone resistance in Escherichia coli (Asp87), Staphylococcus aureus (Glu88), and Campylobacter jejuni (Asp90). J Vet Med Sci, 1994 Aug, 56(4), 697 - 700 A survey of Campylobacter jejuni in broilers from assignment to slaughter using DNA-DNA hybridization; Chuma T et al.; A survey of Campylobacter jejuni in the cecal contents of broilers raised on a farm was carried out by the DNA-DNA hybridization method from the day of assignment to slaughter at about 1-week intervals . C . jejuni was detected in chickens as early as 1 week of age, and was widely detected at each week of age throughout the growing period . In addition, of 20 chickens tested just after assignment, 7 (35%) were C . jejuni positive . It is suggested that newly introduced chickens may have already been contaminated with C . jejuni . The hybridization method was able to detect C . jejuni in the chickens from the day of assignment to 3 weeks of age, but C . jejuni was never detected in the same chickens by the enriched culture method . Therefore, it is suggested that the hybridization method is more sensitive than the enriched culture method. J Clin Microbiol, 1994 Aug, 32(8), 1980 - 5 Enzyme-linked immunosorbent assay for measuring ileal symbiont intracellularis-specific immunoglobulin G response in sera of pigs; Holyoake PK et al.; Proliferative enteritis (PE) is a common intestinal disease on pig farms . The disease is caused by ileal symbiont (IS) intracellularis (Campylobacter-like organisms) bacteria . An enzyme-linked immunosorbent assay (ELISA) was developed to measure IS intracellularis-specific immunoglobulin G (IgG) response in the sera of pigs . The antigen used in the ELISA was filtered, percoll gradient-purified IS intracellularis extracted from the intestines of pigs affected with proliferative hemorrhagic enteropathy . The antibody responses of pigs challenged with intestinal homogenates from pigs affected with proliferative hemorrhagic enteropathy containing IS intracellularis or percoll-gradient purified IS intracellularis were low and variable . The low IgG titers measured in challenged pigs support previous findings that IgG plays a minor role in the immune response of pigs to IS intracellularis . On a farm in which infection was endemic, pigs seroconverted at between 7 and 24 weeks of age . High IgG titers, indicative of maternally acquired antibody, were present in 3-week-old pigs . The IgG titers in piglets were lowest at 6 weeks of age, which approximates the age of onset of clinical disease . These results suggest that IgG plays a role in determining the susceptibilities of pigs to natural infection . Measurements of seroconversion by the ELISA might aid in epidemiological investigations of PE in naturally infected herds . However, the variable antibody responses in experimentally challenged pigs would seem to limit its usefulness as an antemortem diagnostic test for PE. Poult Sci, 1994 Aug, 73(8), 1260 - 6 Cecal carriage of Campylobacter and Salmonella in Dutch broiler flocks at slaughter: a one-year study; Jacobs-Reitsma WF et al.; From March 1992 to March 1993, 187 Dutch broiler flocks were screened to assess their Campylobacter and Salmonella carriage . Every 4 wk at least 10 flocks, at three different slaughterhouses, were screened for presence of these bacteria . Twenty-five cecal samples were taken from each flock . Campylobacter spp . were isolated from 153 out of 187 broiler flocks (82%) . Campylobacter carriage of flocks showed seasonal variation, with the highest contamination rate (100%) during the period June to September and the lowest (50%) in March . Salmonella carriage of the flocks did not show a distinct seasonal variation . Salmonella spp . were isolated from 49 out of 181 broiler flocks (27%) . A positive correlation was found between Campylobacter and Salmonella colonization within flocks . Data on farming conditions and husbandry practices were studied to identify possible risk factors for Campylobacter and Salmonella colonization of Dutch broiler flocks. Enferm Infecc Microbiol Clin, 1994 Aug-Sep, 12(7), 332 - 6 {Campylobacter gastroenteritis in patients with human immunodeficiency virus infection}; Leyes M et al.; BACKGROUND: Campylobacter bacteria are frequent, and usually slight causes of diarrhea in a normal host while in an immunosuppressed host the diarrhea may lead to severe pictures . The aim of this study was to analyze the clinical features of gastroenteritis by Campylobacter spp . in hospitalized seronegative patients and in those with HIV infection . METHODS: A retrospective study of the cases of gastroenteritis by Campylobacter spp . in adult patients admitted in the authors' hospital from January 1988 to July 1993 was carried out . RESULTS: Of the 20 patients studied with gastroenteritis by Campylobacter spp., 13 (65%) had HIV infection . The mean age of the patients was 38 years (range: 18-68 years) with 70% of the cases being males . Seventy seven percent of the HIV positive patients showed diagnostic criteria for AIDS while 71% of the seronegative patients showed a base disease and/or received steroid therapy . The length of the diarrhea was greater in the patients with HIV infection on comparison with the seronegative patients (25 vs . 6 days) . The diarrhea persisted for more than 2 weeks in more than half of the cases of seropositive patients . Fever continued a mean of 24 days in the HIV positive patients as compared with only 5 days in the HIV negative cases . Most of the former patients were treated with erythromycin with good response . Gastroenteritis recurred in one patient and another patient with HIV infection presented a pseudoappendicular picture . No case of bacteremia was detected in either the seropositive or seronegative patients . Campylobacter jejuni was isolated in most of the cases with a high percentage of resistence to quinolone drugs . The mean CD4 lymphocyte count in HIV positive patients was 131/mm3 (range: 1-774) . Mean survival following diagnosis of gastroenteritis by Campylobacter spp . was 8.9 months (range: 1-17) in the patients with AIDS . CONCLUSIONS: Gastroenteritis by Campylobacter spp . in hospitalized patients was related with immunosuppressive states . A clinical profile of prolonged febrile diarrhea was common in HIV positive patients and was associated with a low number of CD4 lymphocytes, advanced HIV infection and short survival. West J Med, 1994 Aug, 161(2), 148 - 52 Clinical aspects of Campylobacter jejuni infections in adults; Peterson MC; Campylobacter jejuni is an almost ubiquitous, microaerophilic, gram-negative rod . Outbreaks have been associated with drinking raw milk or contaminated water and eating poultry . Campylobacter jejuni accounts for 3.2% to 6.1% of cases of diarrheal illness in the general population of the United States, and infected patients frequently present with abdominal pain and fever . Less frequently, C jejuni is responsible for bacteremia, septic arthritis, septic abortion, and other extraintestinal infections . Reactive arthritis, Reiter's syndrome, the Guillain-Barre syndrome, and pancreatitis may accompany or follow C jejuni enterocolitis . Campylobacter jejuni is an important cause of diarrheal illness and is a more commonly identified stool organism than Salmonella or Shigella species . Recurrent and chronic infection is generally reported in immunocompromised hosts. Immun Infekt, 1994 Aug, 22(4), 146 - 8 {Campylobacter fetus subsp . fetus in a patient with liver cirrhosis}; Werner H et al.; Campylobacter fetus subsp . fetus infections are very rare in adults but can occur in immunocompromised elderly people and in patients with chronic disease . We describe the clinical case of a 52-year-old woman with alcoholic liver cirrhosis, where we isolated C.fetus subsp . fetus from blood culture. Dtsch Tierarztl Wochenschr, 1994 Aug, 101(8), 303 - 6 {The distribution and persistence of Campylobacter spp . in chickens}; Glunder G; At the end of the production period, 29 layers each from 70 different flocks were examined for the occurrence of campylobacters in their caecal contents . All flocks were found campylobacter positive . Up to 20 individuals of 13 flocks and 21 to 29 birds of 57 flocks were carriers of Campylobacter spp . Material of the liver was additionally cultured from birds of 36 flocks . The isolation of the organism from livers was possible only from 1 to 6 birds of 16 flocks . In two experiments, chickens were infected at an age of 4 weeks with campylobacter strains inducing different excretion rate . The birds were infected again with a further campylobacter strain at 10 weeks of age . Birds at the same age which were not infected before, served as control . After this superinfection, the excretion of the organisms occurred at the same rate in all groups and did not depend on the strain chosen for the first infection . After an additional second superinfection in the second experiment, the period of excretion of the organism at a high rate was diminished. Intern Med, 1994 Aug, 33(8), 496 - 500 Good's syndrome with a block in the early stage of B cell differentiation and complicated by Campylobacter fetus sepsis; Yamazaki K et al.; A 63-year-old man was admitted for Campylobacter fetus sepsis and immunodeficiency syndrome with thymoma (Good's syndrome) . Serological examination demonstrated hypoimmunoglobulinemia . Analysis of lymphocyte subsets in the peripheral blood and bone marrow showed marked decreases in the proportion of cells bearing B cell markers . However, there were no abnormalities of cellular immunity . This is a rare case of Good's syndrome in Japan in which the pathogenic mechanism involved a block in the early stage of B cell differentiation . Moreover, this is the first case ever reported of Campylobacter fetus sepsis associated with Good's syndrome. Rev Biol Trop, 1994 Aug, 42 Suppl 2, 85 - 92 Ultrastructure of the bacteria Campylobacter and Helicobacter: implications for the phylogeny of mammal gastric bacteria; Hernandez F et al.; Negative staining, thin sections, and surface replica methods were used to produce ultrastructural descriptions of curved-shaped, flagelated bacteria Campylobacter jejuni, Helicobacter pylori, and H . muridarum, the last one from the ileon of mice . The former has nude monotrichous flagella, the others have the sheated lobotrichous type . H . muridarum presents 10-12 periplasmic fibers . A cladogram of some gastric bacteria, based on the evolutionary history of the mammal hosts, hypothesizes that (1) the genus Helicobacter evolved about 65 million years ago and that (2) primate parasites such as H . pylori and H . nemestrinae are close relatives, as are parasites of carnivores which include H . mustelae, H . felis and H . acynonyx. FEMS Microbiol Lett, 1994 Jul 1, 120(1-2), 13 - 7 Characterization of the sat4 gene encoding a streptothricin acetyltransferase in Campylobacter coli BE/G4; Jacob J et al.; The sat4 streptothricin resistance gene from Campylobacter coli BE/G4 was cloned into pUC18, and its nucleotide sequence was determined . Streptothricin acetyltransferase activity was detected in Escherichia coli cells containing recombinant plasmid pAT132 which carries the sat4 gene as an insert . The deduced amino acid sequence displayed 21-27% amino acid identity with streptothricin acetyltransferases from E . coli and streptothricin producers Streptomyces lavendulae and Streptomyces noursei . The sat4 gene was detected by hybridization in clinical and environmental isolates of Campylobacter spp. Tijdschr Diergeneeskd, 1994 Jul 1, 119(13), 390 - 3 {Specific pathogen-free health programs}; Kuiper CJ et al.; SPF programs have been established in various countries, particularly in Denmark and Switzerland these programs are already well advanced . The system proves to be advantageous for export dealings but also with regard to public health . Dalland is an internationally operating Dutch pig breeding organisation . In 1989 a SPF nucleus breeding farm was established in North-East France . A special program was set up for the supply of animals from this SPF nucleus . In this paper it is concluded that SPF can be economically beneficial . Under certain well-described circumstances SPF breeding seems feasible . Its is suggested that in future pigs could be produced that will be free of enteropathogens of interest to public health, e.g . Salmonella, Campylobacter and Yersinia enterocolitica. Infect Immun, 1994 Jul, 62(7), 2687 - 94 Genetic, enzymatic, and pathogenic studies of the iron superoxide dismutase of Campylobacter jejuni; Pesci EC et al.; Campylobacter jejuni is a microaerobic bacterium that produces an acute, self-limiting, watery or bloody diarrhea in humans . Little is known about how C . jejuni causes disease or even what specific capabilities it requires for survival in vivo . The enzyme, superoxide dismutase (SOD), which catalyzes the breakdown of superoxide radicals to hydrogen peroxide and dioxygen is one of the bacterial cell's major defense mechanisms against oxidative damage . A PCR-based search for sod genes in C . jejuni 81-176 revealed that this bacterium contained at least one sod gene . We cloned and sequenced a sod gene from 81-176 and determined that its predicted protein product was most similar to that of FeSODs (sodB genes) . Transcriptional analysis indicated that this gene is monocistronic and may be transcribed from a sigma 70-like promoter . Nondenaturing polyacrylamide gels stained to reveal SOD activities, accompanied by inhibition studies, demonstrated that C . jejuni produces five electrophoretically distinct bands of SOD activity, all of which appeared to be FeSODs . Analysis of an 81-176 sodB strain revealed that all of these FeSOD activities may be products of the one sodB gene that we cloned . The expression and enzymatic activity of the respective sodB and FeSOD produced by both C . jejuni and Helicobacter pylori were examined in Escherichia coli . Both genes were expressed in E . coli, and the proteins produced were enzymatically active . Finally, the ability of the 81-176 sodB strain to survive INT407 cell invasion was found to be significantly decreased (12-fold) compared with that of the parent, suggesting a potential role for SodB in C . jejuni intracellular survival. Zh Mikrobiol Epidemiol Immunobiol, 1994 Jul-Aug, (4), 7 - 10 {The enterotoxigenic activity of Campylobacter isolates taken in acute intestinal infections in children and its clinical assessment}; Gorelov AV et al.; The screening of Campylobacter clinical isolates in ELISA with the use of nitrocellulose filters as solid phase has revealed the possibility of the detection of enterotoxins . The capacity for producing thermolabile enterotoxin has been found in 75.9%, Shiga-like enterotoxin in 56.4% and both enterotoxins in 31.3% of the tested C . jejuni and C . coli strains . The influence of the enterotoxigenic capacity of the strains under study on the severity of Campylobacter infection and on its numerous clinical manifestations in children has been established. Zh Mikrobiol Epidemiol Immunobiol, 1994 Jul-Aug, (4), 32 - 5 {The epidemiological characteristics of single cases of campylobacteriosis in urban inhabitants}; Minaev VI et al.; The results of the analysis of the epidemic situation in Campylobacter infection and the epidemiological markers of the strains of the infective agent have demonstrated that Campylobacter infection, registered as single cases among the residents of Vologda, has an epidemic character . An intensive and uncontrolled process among fowl (chickens) and the absence of effective measures of specific prophylaxis make it necessary that a system of hygienic measures be given priority in the complex of measures aimed at the prophylaxis of Campylobacter infection. Rinsho Shinkeigaku, 1994 Jul, 34(7), 733 - 5 {A patient with Guillain-Barré syndrome in association with Campylobacter jejuni enteritis (PEN19:LIO7) in a patient with HLA-B52 antigen}; Kaida K et al.; We report a 15-year-old boy with Guillain-Barre syndrome (GBS) in association with Campylobacter jejuni enteritis . A neurologic examination revealed distal-dominant weakness and areflexia . Compound muscle action potentials were markedly reduced in amplitude, but the decrease in motor conduction velocities was slight . C . jejuni was isolated from his stool culture and the serotype belonged to PEN19:LIO7 . Thin-layer chromatography-immunostaining showed that his serum IgG reacted strongly with GM1 and weakly with GD1b . The patient had HLA-B52 antigen, whose epitope is very similar to that of B35 antigen. Vet Pathol, 1994 Jul, 31(4), 462 - 7 Specific in situ hybridization of the intracellular organism of porcine proliferative enteropathy; Gebhart CJ et al.; The identity of the intracellular bacteria found in the enterocytes of pigs with proliferative enteropathy was investigated using specific DNA probes to various Campylobacter species and to a novel organism, ileal symbiont intracellularis . The ilea from pigs (Nos . 1-7) that were diagnosed by routine histopathology as having proliferative enteropathy were used . Diagnosis was made on the basis of proliferation of the enterocytes on hematoxylin and eosin-stained sections and the presence of large numbers of intracellular curved organisms on Warthin-Starry silver-stained sections . Four of these pigs (Nos . 1-4) had the chronic form of the disease, porcine intestinal adenomatosis, and three (Nos . 5-7) had the acute form, proliferative hemorrhagic enteropathy . An additional three normal pigs (Nos . 8-10) were obtained from three separate farms with no history of proliferative enteropathy . Frozen ileal sections were examined by in situ hybridization with DNA probes specific for ileal symbiont intracellularis and the three porcine intestinal Campylobacter species, C . coli, C . hyointestinalis, and C . mucosalis . In all seven pigs with either the intestinal adenomatosis or hemorrhagic enteropathy form of the disease, a DNA probe specific for ileal symbiont intracellularis hybridized to localized foci in the apical cytoplasm of ileal enterocytes . These hybridization sites corresponded to the location of intracellular bacteria in silver-stained sections of adjacent tissue . Sections from the three normal pigs tested with this probe and from all pigs tested with the Campylobacter species-specific DNA probes showed no specific hybridization reactions . The identity of the intracellular organism in these diseased pigs is ileal symbiont intracellularis. J Clin Microbiol, 1994 Jul, 32(7), 1718 - 20 Persistence of Campylobacter fetus bacteremia associated with absence of opsonizing antibodies; Neuzil KM et al.; Campylobacter fetus causes systemic infections in immunocompromised hosts . We describe a case in which C . fetus bacteremia apparently relapsed after 7 years in a patient with hypogammaglobulinemia and characterize the serum resistance of the patient's C . fetus strain and the inability of the patient's serum, with and without commercial intravenous immunoglobulin, to opsonize this and another C . fetus strain effectively . The probable presence of a sequestered site of infection in bone, the intrinsic serum resistance of the C . fetus strain, and the absence of specific antibody may account for the persistent infection in this patient . These studies suggest that intravenous immunoglobulin treatment is not useful in eradicating C . fetus bacteremia. East Afr Med J, 1994 Jul, 71(7), 437 - 40 The changing patterns of Campylobacter jejuni/coli in Lagos, Nigeria after ten years; Coker AO et al.; One hundred and forty-five stool samples from children with diarrhoea at various health centres in Lagos were investigated for Campylobacter jejuni/coli . These organisms were isolated from 24 samples (16.5%) . They were later biotyped as 23 C . jejuni and one C . coli . Previous studies (3,6) recorded isolation rates of 5.2% and 11% respectively . No Campylobacter species was isolated from 100 control patients who did not have diarrhoea . Diarrhoea due to Campylobacter jejuni/coli is still common in males, and the ration of males to females is still 2:1 but the isolation rate is now 11.7%:4.8% . All the isolates of Campylobacter jejuni are still from children under two years of age . Campylobacter jejuni biotype 1 accounted for 52.5% in an earlier study (18), but accounted for 87.5% in this study . Serogroup 29(70.8%) was the commonest in this study, whereas serogroup 36(20.7%) was very common in an earlier study(18) . Erythromycin, which is the drug of choice for the treatment of campylobacter enteritis, had 82% sensitivity to the organism(6) but in the present study only 20.8% of the isolates were sensitive to the antibiotic . Betalactamase production was detected in 12.5% of the isolates as against 6.4% earlier reported(17). Eur J Clin Microbiol Infect Dis, 1994 Jul, 13(7), 612 - 5 Development of resistance to macrolide antibiotics in an AIDS patient treated with clarithromycin for Campylobacter jejuni diarrhea; Funke G et al.; In an AIDS patient with diarrhea, identical isolates of Campylobacter jejuni susceptible and, later, resistant to macrolide antibiotics were isolated from feces before and after treatment with clarithromycin . Results of rRNA gene restriction analysis and serotyping suggest that development of resistance rather than simultaneous infection with a susceptible and a resistant strain was responsible for this phenomenon . This is the first report of in vivo development of resistance by Campylobacter jejuni in a patient treated with a macrolide for Campylobacter jejuni infection. Vet Microbiol, 1994 Jul, 41(1-2), 183 - 8 Experimental colonization of mice with Campylobacter jejuni; Berndtson E et al.; The ability of one human and two chicken strains of Campylobacter jejuni to colonise and survive in three different strains of laboratory mice (NMRI, CBA and C57-Black) was studied . Mice were inoculated orally with Campylobacter jejuni and faeces samples were cultured at regular intervals during the following months . The length of colonisation of mice differed between mouse strains but also between Campylobacter strains . The mouse strain C57-Black was not colonised with C . jejuni to the same degree as the other mouse strains . It is concluded that mice can become colonised for prolonged periods and that they may act as reservoirs of Campylobacter for other species. Vet Microbiol, 1994 Jul, 41(1-2), 1 - 9 Reproduction of proliferative enteritis in hamsters with a pure culture of porcine ileal symbiont intracellularis; Jasni S et al.; Hamsters, three weeks old, were inoculated orally with suspensions of intracellular bacteria, grown in tissue culture cells, IEC-18, rat enterocytes . Cells had been infected with suspensions of intracellular bacteria derived from the lesions of proliferative haemorrhagic enteropathy occurring naturally in two pigs 916/91 and 1482/89 . Infected cell lines containing each separate strain, 916/91 and 1482/89, were passaged one, two or five times and pure cultures of intracellular bacteria, identified as ileal symbiont intracellularis by immunological means, were collected from the cells and used as inocula . Ten of sixteen hamsters dosed with 916/91 passaged one or five times, developed lesions of proliferative enteritis evident as necropsy three weeks after inoculation . Hamsters inoculated with 1482/89 passaged twice and stored frozen, or IEC-18 cells alone or those left uninoculated, failed to develop lesions of proliferative enteritis . Campylobacter jejuni infection occurred throughout, in all groups . Marked hyperplasia of ileal enterocytes, associated with numerous intracellular curved bacteria was invariably detected in experimentally affected hamsters . Immunofluorescence reactions with specific antibodies indicated that these intracellular bacteria were also ileal symbiont intracellularis . The results suggested that proliferative enteritis could be reproduced in hamsters with a pure culture of an agent derived from pigs . We concluded that the reproduction of the disease with our inocula containing a single agent clarifies the aetiology of proliferative enteritis in both hamsters and pigs. J Clin Microbiol, 1994 Jul, 32(7), 1788 - 94 Distinct genotypes of human and canine isolates of Campylobacter upsaliensis determined by 16S rRNA gene typing and plasmid profiling; Stanley J et al.; The utility of combined 16S rRNA (rrs) gene restriction fragment length polymorphism and plasmid profiles to differentiate between and within Campylobacter upsaliensis of human and canine origin was examined . Fourteen distinct rrs gene restriction fragment length polymorphs consisting of bands sized between 1.9 and 4.8 kb were observed . The copy number of the 16S rRNA gene was three in most strains of C . upsaliensis . Plasmids were found in almost 60% of the strains; ranging in size from 1.5 to 100 kb, they gave 15 distinct plasmid profiles . All isolates from humans contained one or more plasmids, as did strains isolated from dogs with sporadic diarrhea . The two commonest 16S ribotypes were divided into eight and nine subgroups by plasmid profiling . The genotyping of canine isolates from three veterinary surveys detected both multiple infections and reinfection of dogs . Except for one, each of the isolates from humans constituted a single and unique 16S ribotype, and these more frequently carried plasmids than did canine strains . Ribotypes of human strains were not found among canine isolates . These results suggest that host-specific genotypic differences may exist among strains of C . upsaliensis, for example, intraspecific clones or clone complexes pathogenic for humans. Ann Neurol, 1994 Jun, 35(6), 698 - 703 Monoclonal IgM antibodies to GM1 and asialo-GM1 in chronic neuropathies cross-react with Campylobacter jejuni lipopolysaccharides; Wirguin I et al.; We tested monoclonal IgM anti-GM1 and asialo-GM1 antibodies from 6 patients with chronic motor neuropathies for binding to lipopolysaccharides (LPS) from three stains of Campylobacter jejuni . Four of the 6 patients showed strong reactivity with LPS from at least one of the three C . jejuni strains tested as shown by enzyme-linked immunosorbent assay or western blot . Preabsorption with GM1 or asialo-GM1, or blocking with cholera toxin, prevented antibody binding to LPS . These studies indicate that human anti-GM1 or anti-asialo-GM1 antibodies cross-react with LPS from certain strains of C . jejuni, and that bacterial LPS might provide antigenic stimuli for the activation of B cells expressing anti-GM1 antibodies. J Clin Microbiol, 1994 Jun, 32(6), 1532 - 6 Two-year study of endemic enteric pathogens associated with acute diarrhea in New Caledonia; Germani Y et al.; A longitudinal study of diarrheal disease among patients of all ages with acute diarrhea was carried out in New Caledonia from January 1990 to December 1991 . Stool samples from 2,088 diarrheal patients were examined for parasites, rotavirus, and bacterial pathogens . Potential sources of contamination (drinking water, seawater and bovine and porcine feces) were investigated . One or more enteric pathogens were identified in 41.8 and 40.6% of the persons with diarrhea, in 1990 and 1991, respectively . Salmonella spp., Shigella spp., HEp-2 cell adherent Escherichia coli (diffuse adherent and enteroaggregative), enteropathogenic E . coli (EPEC) (EPEC adherence factor-positive strains belonging to classical serotypes), localized adherent E . coli (non-EPEC), and enterotoxigenic E . coli were the frequently identified enteropathogenic bacteria . Other major enteropathogens were Entamoeba histolytica and Giardia lamblia . Campylobacter jejuni, Clostridium difficile, Clostridium perfringens, Yersinia enterocolitica, and rotavirus were isolated from only a few patients . No Vibrio spp., Aeromonas spp., Plesiomonas spp., Shiga-like-toxin-producing E . coli, enterohemorrhagic E . coli, or enteroinvasive E . coli were identified . Shiga-like toxin I-producing E . coli were present in adult bovines and calves, and heat-stable enterotoxin II-producing enterotoxigenic E . coli were found in pigs. J Clin Microbiol, 1994 Jun, 32(6), 1497 - 502 Rapid detection of Serpulina hyodysenteriae in diagnostic specimens by PCR; Elder RO et al.; A PCR assay for the detection of Serpulina hyodysenteriae in diagnostic specimens was developed on the basis of sequence analysis of a recombinant clone designated pRED3C6 . Clone pRED3C6, which contained a 2.3-kb DNA fragment unique to S . hyodysenteriae, was identified by screening a plasmid library of S . hyodysenteriae isolate B204 genomic DNA in Escherichia coli by colony immunoblot with the mouse monoclonal antibody 10G6/G10, which was produced against cell-free supernatant antigens from the same isolate . Southern blot analysis of HindIII-digested genomic DNA of S . hyodysenteriae serotypes 1 through 7 and of four weakly beta-hemolytic intestinal spirochetes, including Serpulina innocens, with the 2.3-kb DNA fragment of pRED3C6 indicated that the cloned sequence was present exclusively in the seven serotypes of S . hyodysenteriae . An oligonucleotide primer pair for PCR amplification of a 1.55-kb fragment and an internal oligonucleotide probe were designed and synthesized on the basis of sequence analysis of the 2.3-kb DNA fragment of pRED3C6 . Purified genomic DNAs from reference isolates of S . hyodysenteriae serotypes 1 through 9, S . innocens, weakly beta-hemolytic intestinal spirochetes belonging to genotypic groups distinct from those of reference Serpulina spp., other cultivable reference isolates of the order Spirochaetales, and enteric bacteria including Escherichia coli, Salmonella spp., Campylobacter spp., and Bacteroides vulgatus were amplified with the oligonucleotide primer pair in a hot-start PCR . The 1.55-kb products were obtained only in the presence of genomic DNA from each of the nine serotypes of S . hyodysenteriae . The specificity of the 1.55-kb products for S . hyodysenteriae was confirmed on the basis of production of a restriction endonuclease pattern of the PCR products identical to the predicted restriction map analysis of pRED3C6 and positive hybridization signal with the S . hyodysenteriae-specific internal oligonucleotide probe . By using total DNA obtained from normal swine feces inoculated with decreasing concentrations of S . hyodysenteriae cells, the sensitivity of the PCR assay was calculated to be between 1 and 10 organisms per 0.1 g of feces . The PCR assay was 1,000 times more sensitive than conventional culture of dysenteric feces on selective medium . There was complete agreement between the results of PCR assays and anaerobic culture on selective agar medium with diagnostic specimen (n = 9) obtained from six farms on which there were cases with clinical signs suggestive of swine dysentery . Detection of S . hyodysenteriae by PCR amplification of DNA has great potential for rapid identification of S . hyodysenteriae in diagnostic specimens. Biotechniques, 1994 Jun, 16(6), 1064 - 8 Plasmid for the direct subcloning from lambda gt11 to produce an LT-B fusion protein; Meinersmann RJ et al.; We wished to study LT-B fusion proteins for potential as vaccines . We reasoned that a gene that was expressed as a lambda gt11 fusion protein would have a better chance of being expressed fused to LT-B . To facilitate such constructions, we modified plasmid pYA3081 . A BamHI-EcoRI-BamHI* (* is to stop codon) adapter was inserted into the single BamHI site to form a new plasmid-designated pBEB . The insert was designed so that the EcoRI site at the downstream end of the LT-B gene was in the same reading frame as the EcoRI site in the beta-galactosidase gene of lambda gt11 and termination codons occurred downstream in all three reading frames . A clone was selected from a gt11 library based on the production of a protein that reacted with antibodies to flagellin from Campylobacter jejuni . DNA from this clone was digested with EcoRI, the insert fragment purified and ligated into the EcoRI site of pBEB . A fusion protein of 43 kDa was produced that reacted in Western blot with antibodies against LT-B and flagellin . We have succeeded in making an easy method for subcloning fragments from gt11 into a vector for making LT-B fusion proteins. J Okla State Med Assoc, 1994 Jun, 87(6), 267 - 9 Campylobacter fetus presenting as a septic pleural effusion: a case report; Nadir A et al.; The authors describe a rare case of Campylobacter fetus occurring in a patient who showed no overt evidence of immune incompetence . The patient presented with severe cellulitis of the legs, a chronic low grade fever, and a history of recent exposure to sick calves. Appl Environ Microbiol, 1994 Jun, 60(6), 2069 - 75 Inactivation of Escherichia coli O157:H7, salmonellae, and Campylobacter jejuni in raw ground beef by gamma irradiation; Clavero MR et al.; Raw ground beef patties inoculated with stationary-phase cells of Escherichia coli O157:H7, salmonellae, or Campylobacter jejuni were subjected to gamma irradiation (60Co) treatment, with doses ranging from 0 to 2.52 kGy . The influence of two levels of fat (8 to 14% {low fat} and 27 to 28% {high fat}) and temperature (frozen {-17 to -15 degrees C} and refrigerated {3 to 5 degrees C}) on the inactivation of each pathogen by irradiation was investigated . In ascending order of irradiation resistance, the D10 values ranged from 0.175 to 0.235 kGy (C . jejuni), from 0.241 to 0.307 kGy (E . coli O157:H7), and from 0.618 to 0.800 kGy (salmonellae) . Statistical analysis revealed that E . coli O157:H7 had a significantly (P < 0.05) higher D10 value when irradiated at -17 to -15 degrees C than when irradiated at 3 to 5 degrees C . Regardless of the temperature during irradiation, the level of fat did not have a significant effect on the D10 value . Salmonellae behaved like E . coli O157:H7 in low-fat beef, but temperature did not have a significant effect when the pathogen was irradiated in high-fat ground beef . Significantly higher D10 values were calculated for C . jejuni irradiated in frozen than in refrigerated low-fat beef . C . jejuni was more resistant to irradiation in low-fat beef than in high-fat beef when treatment was at -17 to -15 degrees C . Regardless of the fat level and temperature during inactivation, these pathogens were highly sensitive to gamma irradiation.(ABSTRACT TRUNCATED AT 250 WORDS) Gut, 1994 Jun, 35(6), 755 - 7 Effects of Helicobacter pylori on histamine and carbachol stimulated acid secretion by human parietal cells; Jablonowski H et al.; Helicobacter pylori (H pylori) infection is associated with hypo, normal, and hypersecretory disorders of the gastric mucosa . Pathophysiological pathways by which H pylori interacts with acid secretion are still unclear . The effects of H pylori on (14C) aminopyrine uptake by human parietal cells were examined as an indirect assay for acid secretion . Isolated oxyntic glands were stimulated with submaximal concentrations of histamine or carbachol and incubated with sonicates of different H pylori strains . Omeprazole and sonicates of Campylobacter jejuni served as positive and negative controls, respectively . Two of four H pylori strains reduced hydrochloric acid sequestration within the parietal cells significantly and in a dose dependent manner in up to 80% . Interaction with acid secretion may therefore constitute a factor contributing to a distinct pathogenicity of H pylori strains. Scand J Urol Nephrol, 1994 Jun, 28(2), 179 - 81 Henoch-Schönlein purpura associated with Campylobacter jejuni enteritis . Case report; Lind KM et al.; After an episode of enteritis caused by C . jejuni a 15-year-old boy developed Schonlein-Henoch purpura . Because of continuous proteinuria a renal biopsy was performed . Light microscopy revealed focal proliferative glomerulonephritis . IgA was the dominant immunoglobulin . A causal relationship between enteritis caused by C . jejuni and Schonlein-Henoch purpura complicated with focal proliferative glomerulonephritis is suggested. Oral Microbiol Immunol, 1994 Jun, 9(3), 136 - 41 The oral gram-negative anaerobic microflora in young children: longitudinal changes from edentulous to dentate mouth; Kononen E et al.; Eruption of primary teeth has a great influence on the oral environment by providing suitable niches for bacterial colonization . The composition of oral gram-negative anaerobic microflora was investigated in 21 young children (mean age 32 months) with primary dentition . The bacterial findings of samples were compared with those of the same children collected at their edentulous infant period (mean age 3 months) . During the primary period, 2 samples were collected from each child: a sample with dental floss from gingival margin of 2 teeth and stimulated saliva pooled with a mucosal swab sample . Both samples were cultured aerobically and anaerobically using nonselective and selective media . Prevotella melaninogenica, nonpigmented Prevotella spp., Fusobacterium nucleatum group and Capnocytophaga spp . were found in all children at the older age, whereas they occurred in edentulous mouth in 76%, 62%, 67% and 19%, respectively . The occurrence of Prevotella loescheii increased from 14% to 90%, Prevotella intermedia from 10% to 67%, Leptotrichia spp . from 24 to 71%, Campylobacter (Wolinella) spp . from 5 to 43% and Eikenella corrodens from 5 to 57% . Only the occurrence of Bacteroides gracilis and Veillonella spp . remained at about the same level . Species not isolated from the edentulous mouth, such as Prevotella denticola, Fusobacterium spp . other than the F . nucleatum group and Selenomonas spp . were now detected in 71%, 71% and 43% of the children . The stability of the colonizing P . melaninogenica strain(s) in the oral cavity was determined by using ribotyping; 1-2 isolates per child from the edentulous infant period of 9 children and 3-15 isolates per child from their primary dentition period were analyzed.(ABSTRACT TRUNCATED AT 250 WORDS) East Afr Med J, 1994 Jun, 71(6), 376 - 8 Cryptosporidiosis among medical patients with the acquired immunodeficiency syndrome in Tikur Anbessa Teaching Hospital, Ethiopia; Mengesha B; Fresh stool specimens, collected at random from 63 medical in-patients with acquired immunodeficiency syndrome (AIDS), were studied prospectively for Cryptosporidium oocyst . The diagnosis of AIDS was made according to the clinical case definition of the Bangui criteria . These patients presented with profuse watery diarrhoea, significant weight loss and other associated symptoms and signs of clinical manifestations of symptomatic human immunodeficiency virus (HIV) infection . Using the modified Kinyoun acid fast staining technique, 25(39.7%) of the stool specimens were positive for Cryptosporidium oocyst . This study showed that the protozoan, Cryptosporidium parvum, may be responsible for a significant proportion of cases of chronic diarrhoea among AIDS patients in EthiopiaPIP: All adult patients who were admitted into hospital with chronic diarrhea and who fulfilled the modified World Health Organization surveillance case definition for AIDS (Bangui definition) were included in this study . The presence of generalized Kaposi's sarcoma or cryptococcal meningitis are sufficient by themselves for the diagnosis of AIDS surveillance purposes . Freshly passed liquid stool was collected from each patient . There were a total of 63 patients, 40 males and 23 females, 18-49 years old with a mean of 31 years . 42 (75%) of the 63 were residents of Addis Ababa city . 20 (31%) patients died in hospital . The duration of diarrhea varied from just a month to 9 months, with a mean of 4 months . The stool was watery in the majority of the patients except in those with amoebiasis and bacterial enteropathogen in whom blood and/or mucus was also present . Weight loss was dramatic and reached as much as 30-40% of body weight in some . 25 (39.7%) of study patients had oocyst of Cryptosporidium in their stools . Stool tests for other ova and parasites showed 42 (77%) to be negative . 21 (33%) were positive for single and mixed infections . Entamoeba histolytica accounted for 15 (11%), while Giardia lamblia and Strongyloides stercolaris were positive in 3 patients . Ascaris lumbricoides and hookworm were present concomitantly in 6 of the cases with protozoal parasites . 21 patients had stool cultures that grew no enteropathogens . In 6 others, cultures grew Salmonella, Shigella and Campylobacter species in 4, 2, and 2 respectively . Double infections were observed in 2 cases . Enteropathogenic parasites and microbial agents were identified in about 63% of patients . In the absence of effective antibiotic or chemotherapeutic agents for the treatment of protozoan enteropathogen, the stool smear for Cryptosporidium adds extra cost to the management of AIDS . Prevention remains the only alternative, both for AIDS and cryptosporidiosis . Onderstepoort J Vet Res, 1994 Jun, 61(2), 193 - 5 First isolation of Campylobacter jejuni from the vaginal discharge of three bitches after abortion in South Africa; Odendaal MW et al.; Campylobacter jejuni was isolated in pure culture from the vaginal discharge from three German Shepherd bitches after late-pregnancy abortions . The main clinical sign occurring in the bitches was a profuse and odourless haemorrhagic vaginal discharge. J Bacteriol, 1994 Jun, 176(11), 3303 - 13 Structural and antigenic characteristics of Campylobacter coli FlaA flagellin; Power ME et al.; The polar flagellar filament of Campylobacter coli VC167 is composed of two highly related (98%) flagellin subunit proteins, FlaA and FlaB, whose antigenic specificities result from posttranslational modification . FlaA is the predominant flagellin species, and mutants expressing only FlaA form a full-length flagellar filament . Although the deduced M(r) of type 2 (T2) FlaA is 58,884 and the apparent M(r) by sodium dodecyl sulfate-polyacrylamide gel electrophoresis is 59,500, the solution weight-average M(r) by sedimentation analysis was 63,000 . Circular dichroism studies in the presence or absence of 0.1% sodium dodecyl sulfate or 50% trifluorethanol showed that the secondary structure of T2 FlaA flagellin was altered, with alpha-helix structure being increased to 25% in the nonpolar environment . The molecule also contained 35 to 48% beta-sheet and 11 to 29% beta-turn structure . Mimeotope analysis of octapeptides representing the sequence of FlaA together with immunoelectron microscopy and enzyme-linked immunosorbent assay with a panel of antisera indicated that many residues in presumed linear epitopes were inaccessible or nonepitopic in the assembled filament, with the majority being in the N-terminal 337 residues of the 572-residue flagellin . Residues at the carboxy-terminal end of the T2 FlaA subunit also become inaccessible upon assembly . Digestion with trypsin, chymotrypsin, and endoproteinase Glu-C revealed a protease-resistant domain with an approximate M(r) of 18,700 between residues 193 and 375 . Digestion with endoproteinase Arg-C and endoproteinase Lys-C allowed the mapping of a segment of surface-exposed FlaA sequence which contributes serospecificity to the VC167 T2 flagellar filament at residues between 421 and 480. Rev Prat, 1994 May 15, 44(10), 1333 - 8 {New bacterial infections in the course of AIDS}; Rousseau MC et al.; The authors reviewed the recent data on unusual or newly described bacterial infection in AIDS . Mycoplasma species are frequently associated with AIDS and hypothesized to play as a cofactor in AIDS evolution (Mycoplasma fermentans and Mycoplasma penetrans) . Rochalimea henselea and Rochalimea quintana are the agents of peliosis and bacillary angiomatosis . Some Mycobacterium and Campylobacter species, not previously considered as human pathogens, have been recently involved in severe infection in patients with AIDS. Tidsskr Nor Laegeforen, 1994 May 10, 114(12), 1416 - 20 {Diagnosis and treatment of infections of the digestive system in HIV-infected patients}; Oktedalen O et al.; Patients with HIV infection often suffer from opportunistic and bacterial infections of the digestive tract . The most common agents are Candida albicans, Herpes simplex and Cytomegalovirus, Mycobacterium avium intracellulare, Cryptosporidium parvum, and enteropathogenic bacteria such as Salmonella, Shigella and Campylobacter . The diagnosis is established by means of microbiological examination of blood and faeces, often supplemented by gastrointestinal endoscopy, with mucosal biopsies for culture and histology . Most patients respond well to specific treatment, but the infections tend to relapse after withdrawal of drugs. Infect Immun, 1994 May, 62(5), 2122 - 5 Lipopolysaccharides from Campylobacter jejuni associated with Guillain-Barré syndrome patients mimic human gangliosides in structure; Aspinall GO et al.; Lipopolysaccharides extracted from Campylobacter jejuni serostrains (serotype reference strains) for serotypes O:4 and O:19 were found to have core oligosaccharides with terminal structures resembling human gangliosides GM1 and GD1a . High-molecular-weight molecules that reflected the presence of O chains were shown in immunoblots to be immunologically specific for each serostrain . The O:19 antiserum also reacted strongly with core oligosaccharides of two isolates from patients with Guillain-Barre syndrome (GBS), but the banding patterns and molecular structures were different from those of the O:19 serostrain . A neuraminobiose disaccharide unit is attached to the terminal Gal residue in one isolate, and the other isolate lacked terminal N-acetyl glucosamine and galactose with attached sialic acid so that the sialic acid residues were present in a neuraminobiose unit linked to the only remaining galactose . Analysis of the high-M(r) lipopolysaccharides of the O:19 serostrain and the two isolates from GBS patients revealed the presence of a hyaluronic acid-like polymer with disaccharide-repeating units consisting of beta-D-glucuronic acid amidated with 2-amino-2-deoxyglycerol and N-acetyl glucosamine . The results confirm a potential role for the core oligosaccharides in the etiology of GBS but also suggest that the O-chain polysaccharide may be a contributing factor. Arkh Patol, 1994 May-Jun, 56(3), 27 - 30 {Acute Campylobacter colitis}; Shalygina NB et al.; Biopsies of colon mucous membrane obtained at colonofibroscopy of 16 adult patients with colon campylobacteriosis were studied . Acute diffuse colitis was found in all cases . Exudative inflammation with hemorrhagic component and microerosion formation was found up to day 10 of the disease . Catarrhal inflammation was observed from day 10-16 during the period of convalescence . Abundant infiltration of tunica propria with eosinophylic leucocytes is a peculiar feature of colon campylobacteriosis . This, together with a diffuse character of inflammation, makes a difference with Shigella colitis . Complete normalization of the mucosa does not occur within 16 days. Int J Food Microbiol, 1994 May, 22(2-3), 189 - 200 Simple extraction of Campylobacter lipopolysaccharide and protein antigens and production of their antibodies in egg yolk; Chandan V et al.; Antigens were heat extracted from Campylobacter jejuni (LI04) and C . coli (LI020) in the presence of ethylenediaminetetraacetate (EDTA) and were recovered in the supernatant of a low-speed centrifugation . The method is simpler, safer and more efficient in extracting lipopolysaccharide (LPS) antigens than the hot phenol method . The extracted antigens (LPS plus several proteins) elicited production of antigen-specific antibodies in the egg yolk of immunized hens . Antibodies purified by polyethyleneglycol fractionation were used to detect antigens fractionated on SDS polyacrylamide gel electrophoresis. Aust Vet J, 1994 May, 71(5), 140 - 3 Diagnosis of bovine venereal campylobacteriosis by ELISA; Hum S et al.; An enzyme-linked immunosorbent assay (ELISA) measuring IgA antibodies in the vaginal mucus was used to diagnose bovine venereal campylobacteriosis in 241 herds with infertility and abortions . The presence of the disease was confirmed on 84 farms (34.8%) and it was suspected on a further 27 farms (11.2%) . The specificity of the ELISA was found to be 98.5% but in the absence of a reliable comparative test sensitivity can not be estimated . Vaccination against campylobacteriosis will not interfere with the IgA ELISA because only IgG is present in the vaginal mucus of vaccinates . Because of the possibility of false reactions caused by antibody fluctuations in individual cattle, the ELISA is best used as a herd test . It appears that at present the vaginal mucus IgA ELISA is the test of choice for the diagnosis of bovine venereal campylobacteriosis. J Gastroenterol Hepatol, 1994 May-Jun, 9(3), 291 - 303 AIDS and the gut; Chui DW et al.; There are increasing challenges for the practising gastroenterologist in treating AIDS-related gastrointestinal diseases . The differential diagnoses of dysphagia and odynophagia include cytomegalovirus (CMV) and herpes simplex virus (HSV) infection, non-specific aphthous ulceration and non-AIDS oesophageal diseases, especially reflux oesophagitis . Chronic subacute abdominal pain with nausea, vomiting, early satiety and weight loss is suggestive of an obstructive lesion caused by lymphoma or Kaposi's sarcoma . Severe acute abdominal pain can indicate pancreatitis or intestinal perforation due to cytomegalovirus . Right upper quadrant pain (with or without fever, vomiting or abnormal liver function tests with a cholestatic profile) is suggestive of hepatobiliary pathology including cholecystitis, cholangitis, acalculous cholecystitis and AIDS cholangiopathy . Diarrhoea is the most common gastrointestinal symptom of AIDS, affecting 50-90% of patients . Causes of AIDS diarrhoea include protozoa (Cryptosporidium parvum, Isospora belli, Enterocytozoon bieneusi, Septata intestinalis, Cyclospora spp, Entamoeba histolytica and Giardia lamblia), bacteria (Mycobacterium avium-intracellulare, Clostridium difficile, Salmonella, Shigella and Campylobacter jejuni), and viruses (CMV, HSV and possibly HIV) . Chronic diarrhoea, malnutrition and weight loss can shorten the life-span of patients with AIDS . Elemental diets, isotonic formulas, medium chain triglycerides and total parenteral nutrition have been tried with little success in AIDS patients with severe diarrhoea and wasting. J Clin Microbiol, 1994 May, 32(5), 1229 - 37 Intracellular Campylobacter-like organism from ferrets and hamsters with proliferative bowel disease is a Desulfovibrio sp; Fox JG et al.; Proliferative bowel disease is an intestinal disorder of a variety of domestic animals associated with the presence of an intracellular Campylobacter-like organism (ICLO) . We have identified the ICLO obtained from a ferret with proliferative colitis by 16S rRNA sequence analysis . In this ferret, proliferative bowel tissue containing the ICLO had translocated to the mesenteric lymph nodes, omentum, and liver . The 16S rRNA genes of the ICLO were amplified from an infected fragment of extraintestinal tissue by using universal prokaryotic primers . Approximately 1,480 bases of the amplified 16S rRNA gene were sequenced by cycle sequencing . Comparison of the sequence of the ICLO with those of over 400 bacteria in our data base indicated that the sequence of the ICLO was most closely related to that of Desulfovibrio desulfuricans (87.5% similarity) . Phylogenetic analysis with 12 Desulfovibrio species and 20 species from related genera placed the ICLO in a subcluster within the genus Desulfovibrio with D . desulfuricans and 5 other Desulfovibrio species . We will refer to this organism as the intracellular Desulfovibrio organism (IDO) . Specific primers were produced for PCR amplification of a 550-base fragment of the 16S rRNA gene of the IDO in proliferative intestinal tissue samples . This unique 550-base segment was amplified from samples of frozen intestinal tissue from nine ferrets and three hamsters with ICLO-associated disease but not in four intestinal tissue samples from animals without the ICLO-associated disease . The 550-base amplified products from the bowel tissues of one hamster and one ferret were fully sequenced . The ferret IDO partial sequence was identical to the previously determined 16S rRNA sequence over its length, and the hamster IDO sequence differed by a single base . The same intracellular organism has been identified in proliferative intestinal tissues of swine and that the organism has been successfully maintained in tissue culture . The availability of specific primers for PCR-based detection of this intracellular Desulfovibrio organism will aid in the determination of its role in the pathogenesis of proliferative bowel disease in a variety of infected hosts. Microbiology, 1994 May, 140 ( Pt 5), 1203 - 8 Cloning, nucleotide sequence and characterization of a gene encoding superoxide dismutase from Campylobacter jejuni and Campylobacter coli; Purdy D et al.; Genes encoding superoxide dismutase (SOD: EC 1.15.1.1) were cloned from Campylobacter jejuni NCTC 11351 and Campylobacter coli UA585 by heterologous complementation of a SOD-deficient Escherichia coli mutant . Deletion analysis of the cloned C . jejuni DNA assigned the sod gene to a 1.2 kb insert and this contained an open reading frame of 660 bp . The deduced gene product of 220 amino acids was 71% identical to the E . coli iron-containing SOD and 60% identical to the E . coli manganese-containing SOD . The recombinant SOD was expressed at high levels in E . coli and protected a sodA sodB double mutant from the toxic effects of methyl viologen . Nucleotide sequence analysis of the corresponding gene from C . coli showed it to be 92% identical to that from C . jejuni. Rev Invest Clin, 1994 May-Jun, 46(3), 221 - 9 {Biochemical characterization of Campylobacter using the "Micro Campy" method}; Granados Silvestre MA et al.; Three species of Campylobacter are recognized as enteropathogens in children under five years of age and in immunocompromised patients . Several groups have tried to classify them using biochemical profiles and have found six different patterns which comprise 12 individual tests . We have designed a "Micro Campy" method which identifies C . jejuni, C . coli and C . lari classifying them into several biotypes . Fifty two Campylobacter strains isolated from humans were studied and classified in 24 different patterns . Strains isolated from diarrhea episodes were seen in two specific patterns . We propose the use of this easy to perform method which has a reproducibility higher than 92% in biotyping Campylobacter strains. Mikrobiol Z, 1994 May-Jun, 56(3), 51 - 4 {The fatty acid composition of the total lipids in bacteria of the genus Campylobacter}; Kirik DL et al.; Composition of fatty acids of total lipids in home strains of campylobacteria has been studied . Lipids of all the strains of C . jejuni and C . coli mainly consist of saturated fatty acids (from 75.7 to 78.7%) with predominance of tetradecanoic and hexadecanoic fatty acids . The level of unsaturated fatty acids is considerably lower (from 21.0 to 22.5%); These acids are mainly presented by hexadecene acid . Qualitative composition of fatty acids of total lipids in C . jejuni and C . coli does not permit using it for differentiation within these species. Infect Immun, 1994 May, 62(5), 2101 - 3 Penner's serotype 4 of Campylobacter jejuni has a lipopolysaccharide that bears a GM1 ganglioside epitope as well as one that bears a GD1 a epitope; Yuki N et al.; The carbohydrate structures of lipopolysaccharides (LPSs) of Campylobacter jejuni strains belonging to Penner's serotypes (PEN) 1, 2, 4, 19, 23, and 36 were studied by thin-layer chromatography and immunostaining with several monoclonal antiganglioside antibodies . Anti-GM1 and anti-GD1a antibodies reacted with the LPSs of PEN 1, 4, and 19 . Aspinall et al . (G . O . Aspinall, A . G . McDonald, T . S . Raju, H . Pang, A . P . Moran, and J . L . Penner . Eur . J . Biochem . 213:1017-1027, 1993) recently reported that the LPS of PEN 4 has a GD1a ganglioside-like structure rather than a GM1-like structure . We found that the LPS fraction of C . jejuni (PEN 4) has an LPS that bears a GM1 epitope as well as an LPS that bears a GD1a epitope. Clin Diagn Lab Immunol, 1994 May, 1(3), 310 - 7 Serum antibody response to the superficial and released components of Helicobacter pylori; Bazillou M et al.; Superficial and released components were extracted from six selected Helicobacter pylori strains . The protein and antigenic profiles of these extracts were representative of the profiles found most frequently among the clinical strains and included major peptidic fractions at 19, 23.5, 57, 68, 76, 118, and 132 kDa and major antigens at 68, 57, and 23.5 kDa . Immuno-cross-reactions were seen with a hyperimmune rabbit serum to Campylobacter fetus but not with sera to Campylobacter jejuni or Salmonella spp . An antigenic preparation was obtained by pooling equivalent quantities of each extract, and the antigenic preparation was used to study the antibody responses of sera from 65 French patients and 127 Tunisian patients . By enzyme-linked immunosorbent assay, we observed that the sera from French and Tunisian patients clustered into two populations, defined as antibody positive (72 patients) and antibody negative (120 patients) . The antibody-positive patients were more frequently infected with H . pylori (P < 0.01) and were more frequently affected with gastritis (P = 0.05) . However, no correlation between antibody levels and clinical signs of dyspepsia was noticed . The proportions of antibody-positive patients were similar in France and Tunisia . Antibody-positive and antibody-negative sera were studied by western blot (immunoblot) analysis . The antibody-positive sera revealed an average of 7.7 antigenic bands, whereas the antibody-negative sera revealed an average of 2.4 antigenic bands (P < 0.01) . The antigens between 15 and 40 kDa and greater than 66 kDa were specifically recognized by the antibody-positive sera, although in this molecular size range the antibody profiles of these sera exhibited a fairly high degree of diversity . We conclude that the superficial and released components from H . pylori contain a variety of bacterial immunogens and may be useful in antigenic preparations for the serodiagnosis of H . pylori infections . Moreover, a group of antigens in combination appears to be useful for discriminating antibody-positive and antibody-negative patients. J Periodontol, 1994 Apr, 65(4), 357 - 63 Impaired bactericidal activity of PMN from two brothers with necrotizing ulcerative gingivo-periodontitis; Cutler CW et al.; The purpose of this study was to investigate the pathogenesis of necrotizing ulcerative gingivo-periodontitis (ANUP) diagnosed in two brothers, age 9 (ANUP1) and 14 (ANUP2) from rural Egypt . Complete blood count, differential and blood chemistry were within normal limits for both brothers and they were not malnourished . The phagocytosis and killing function of their polymorphonuclear leukocytes (PMN) towards four bacterial species were assessed using a fluorochrome microassay . The selection of bacterial species was based on preliminary microbiological results in early onset periodontitis in Egypt . Fluorochrome-labeled Prevotella intermedia, Peptostreptococcus micros, Campylobacter rectus, and Porphyromonas gingivalis were pre-opsonized with ANUP serum and added to PMN from both ANUP patients, as well as PMN from three sex-matched and two sex- and age-matched healthy Egyptian control (CTL) subjects . We found significant depressions (P < 0.05) in PMN phagocytosis and killing of C . rectus and P . intermedia by ANUP1 and ANUP2, when compared to all CTL PMN . An assessment of the Gram-negative subgingival microflora present in both ANUP patients (in colony forming unit percent of total CFU recovered) (CFU %) revealed the presence of P . intermedia (ANUP1, 41.7 CFU %; ANUP2, 14.8 CFU %), Fusobacterium nucleatum (ANUP1, 3.6 CFU %; ANUP2, 48.1 CFU %), and Veillonella spp . (ANUP1, 18.2 CFU %; ANUP2, 18.5 CFU %) . Spirochetes were also observed in cytocentrifuged, Gram-stained plaque from both ANUP patients . The predominant Gram-positive bacterial species recovered from both NUG1 and NUG2 was Streptococcus morbillorum. J Periodontol, 1994 Apr, 65(4), 309 - 15 Flow-cytometric identification and detection of Porphyromonas gingivalis by a LPS specific monoclonal antibody; Kamiya I et al.; The purpose of this study was to identify Porphyromonas gingivalis (P . gingivalis) by flow cytometry (FCM) using a monoclonal antibody (MAb) OMR-Bg1E directed to P . gingivalis-specific lipopolysaccharide (LPS) . The P . gingivalis strains ATCC 33277, 381, ESO75, W50, and A7A1 were selected for the study . Fusobacterium nucleatum (F . nucleatum), Prevotella intermedia (P . intermedia), Campylobacter rectus (C . rectus), Streptococcus sanguis (S . sanguis) and Actinobacillus actinomycetemcomitans (A . actinomycetemcomitans) served as controls . A suspension of 10(7) bacteria/ml of each bacteria was prepared and then reacted with a P . gingivalis specific MAb OMR-Bg1E and fluorescein isothiocyanate (FITC) labeled second antibody . These samples were analyzed by FCM . Bacterial specific binding aggregate on data was separated out by the forward- and side-angle-scatter characteristics, while non-specific binding (NSB) was eliminated by excluding the region with mouse IgG-positive and second antibody-positive area . FCM detected a mean range of 56.2% to 97.2% P . gingivalis strains . There was a 5.1% non-specific binding using FCM to non-P . gingivalis strains . When the P . gingivalis concentration was adjusted to 10(2), 10(4), and 10(6) bacteria/ml, a detection rate of 35.7%, 48.1%, and 91.4%, was respectively observed . The lower sensitivity of the flow cytometric assay was 10(2) bacteria/ml . When P . gingivalis was added to P . intermedia suspension at 1, 20, 40, 60, and 80%, the MAb-positive fraction yielded by FCM displayed a coefficient of determination of 0.967 with the actual percentage of P . gingivalis and could be regressed to a linear function.(ABSTRACT TRUNCATED AT 250 WORDS) Arch Pathol Lab Med, 1994 Apr, 118(4), 350 - 65 Bacteria in blood for transfusion . A review; Sazama K; OBJECTIVE: To summarize reports of bacterial contamination of blood components for transfusion during this century, considering implicated microorganisms and patient outcomes, with identification and discussion of recommended methods to reduce or eliminate this problem . DATA SOURCES: Articles published in the English-language literature from which summary tables of all reported bacterial infections were prepared by collating published case reports, including fatalities, emphasizing recent concerns about Yersinia contamination . STUDY SELECTION: All case reports and related review articles relevant to issues about bacteria implicated in transfusion-associated sepsis were included . DATA EXTRACTION: The author personally extracted all data . DATA SYNTHESIS: With the exception of Yersinia species and Campylobacter jejuni in red blood cells and Salmonella heidelburg in platelets, the majority of bacteria implicated continue to be those found in the environment and as normal skin flora . Existing measures to identify contaminated components before transfusing them are inadequate . Manufacturing efforts to sterilize these components by various methods may result in greater benefit than those directed toward preventing contamination during collections . CONCLUSIONS: Bacterial contamination remains a problem for transfusion medicine . Active research should continue to focus on elimination of contaminants by filtration, chemical additives, or irradiation, as well as innovative measures to detect and exclude infected units from transfusion. Am J Clin Pathol, 1994 Apr, 101(4 Suppl 1), S14 - 7 Practical considerations in the laboratory diagnosis of bacterial enteric infections; Nolte FS; Diarrheal diseases caused by bacterial pathogens are important causes of morbidity in the United States, and considerable laboratory resources are spent to detect these enteric pathogens . This article reviews recent developments in the detection and identification of Campylobacter spp, enterohemorrhagic Escherichia coli, and Clostridium difficile, and outlines cost-effective strategies for use of stool cultures. Epidemiol Infect, 1994 Apr, 112(2), 359 - 65 Immunoglobulin A antibodies directed against Campylobacter jejuni flagellin present in breast-milk; Nachamkin I et al.; We studied the relationship between IgA anti-campylobacter flagellin antibodies in breast milk samples and protection of breastfed infants living in a rural Mexican village from campylobacter infection . There were fewer episodes of campylobacter infection (symptomatic and asymptomatic combined) in infants breastfed with milk containing specific anti-flagellin antibodies (1.2/child/year, 95% CI 0.6-1.8) versus non-breastfed children (3.3/child/year, 95% CI 1.8-4.8; P < 0.01) . Infants breastfed with milk that was anti-flagellin antibody negative by ELISA also had fewer episodes of infection compared with non-breastfed children, but the difference did not reach statistical significance (1.8/child/year, 95% CI 0.7-3.0 versus 3.3/child/year, 95% CI 1.8-4.8, P > 0.05) . Breastfeeding has a protective effect against campylobacter infection and is associated with the presence of specific antibodies directed against campylobacter flagellin. Epidemiol Infect, 1994 Apr, 112(2), 347 - 57 False positive legionella serology in campylobacter infection: campylobacter serotypes, duration of antibody response and elimination of cross-reactions in the indirect fluorescent antibody test; Marshall LE et al.; Sera from 83 patients with campylobacter gastroenteritis were examined for the presence of legionella antibodies by indirect immunofluorescence . Twenty-one patients (25%) had positive titres (> or = 16) including 11 patients with titres of > or = 128 . Legionella seropositivity persisted in 5 of 9 patients (55%) studied for 6-9 months . Campylobacter isolates were serotyped by the Penner scheme . Isolates associated with legionella seropositivity included Penner types 1, 2 and 4, the common endemic serotypes in England . Campylobacter blocking fluids were prepared from a range of Penner reference strains . The blocking fluid prepared from Penner type 11 was the most efficient at inhibiting the false-positive legionella titres . Using this absorption step legionella titres were inhibited from 24 of 26 patients (92%) with campylobacter but not from 8 patients with culture-proven legionnaires' disease . We recommend that this method is incorporated into routine diagnostic legionella serology in order to eliminate false-positive reactions due to campylobacter. J Bacteriol, 1994 Apr, 176(7), 1865 - 71 Cloning, characterization, and nucleotide sequence analysis of the argH gene from Campylobacter jejuni TGH9011 encoding argininosuccinate lyase; Hani EK et al.; The complete structural gene for argininosuccinate lyase (argH) from Campylobacter jejuni TGH9011 has been cloned into Escherichia coli by complementation of an E . coli argH auxotrophic mutant . The gene has been subcloned for sequencing on a 4.1-kb DNA segment and localized by the complementing activity of deletion mutants . The complete DNA sequence of the C . jejuni argH gene was determined . The transcription start point for argH mRNA was determined by primer extension analysis and found to be within the coding sequence of the upstream gene, identified as the phosphoenolpyruvate carboxykinase gene (ppc) . The argininosuccinate lyase and the phosphoenolpyruvate carboxykinase reading frames overlap by one base, the second example of this phenomenon in C . jejuni chromosomal genes . The enzyme has a deduced subunit molecular weight of 51,831 . Recombinant plasmids containing the argH gene generate a 56-kDa protein and a 43-kDa protein in E . coli maxicells . An alternate translation initiation producing a polypeptide with a deduced molecular mass of 42 kDa may account for the smaller protein observed in sodium dodecyl sulfate-polyacrylamide gel electrophoresis . The C . jejuni argH gene shows nucleotide homology to both yeast and human argininosuccinate lyase genes, and conserved amino acid domains are evident between the corresponding proteins. J Infect Dis, 1994 Apr, 169(4), 916 - 9 Etiology of diarrhea in a rural community in western Thailand: importance of enteric viruses and enterovirulent Escherichia coli; Echeverria P et al.; The etiology of gastroenteritis was determined in children and adults with diarrhea seen at a district hospital and three government health clinics in Suan Phung, western Thailand, in 1991 . Enteric viruses (rotavirus and astrovirus) were identified in 40%, shigellae in 18%, attaching and effacing Escherichia coli in 13%, Campylobacter jejuni in 9%, and enterotoxigenic E . coli in 7% of children < 5 years old with diarrhea seen at the hospital . Enteric viruses were detected in 15% (24/156) of patients with diarrhea > or = 5 years old and were the only enteric pathogens identified in 12 patients ages 7-79 years (2 astrovirus, 10 rotavirus infections) . Attaching and effacing E . coli, rotavirus, and astrovirus were potential causes of diarrhea in children and adults in this population. Ann Trop Med Parasitol, 1994 Apr, 88(2), 175 - 82 Diarrhoeal disease in children in Gaza; Sallon S et al.; A 1-year prospective study in Gaza of diarrhoeal disease in children aged < 5 years demonstrated that Salmonella spp . (18.5% of cases), Cryptosporidium (14.6%), Campylobacter spp . (8.3%) and rotavirus (6.8%) were the major pathogens . However, when compared with non-diarrhoeic controls, only Cryptosporidium and rotavirus were significantly associated with diarrhoea . Cryptosporidiosis was found only in children aged < 2 years and significantly more children with cryptosporidiosis were malnourished . This malnutrition may have been due to the infection, since children with cryptosporidiosis tended to have had diarrhoea for relatively long periods prior to admission . It was not possible to distinguish between the different enteropathogens on clinical grounds . However, more children with rotavirus infection vomited and cryptosporidial diarrhoea lasted significantly longer (14.9 days) than rotavirus diarrhoea (5.9 days) . Overcrowding was linked with an increased risk of cryptosporidiosis and breast feeding was associated with some protection . Twenty-one of the 29 children who died during the study died with diarrhoea and Cryptosporidium was detected in eight (38%) of these 21 children. J Endod, 1994 Apr, 20(4), 169 - 72 Bacteria invading periapical cementum; Kiryu T et al.; The aim of this study was to determine whether microorganisms invade periapical cementum of human teeth from the adjacent periapical lesions . We therefore attempted to isolate microorganisms from periapical cementum through the adoption of standard anaerobic procedures for obligate anaerobes . Samples of cementum were taken from 10 amputated tooth roots at the time of apicoectomy . From two of these samples, bacteria were recovered after anaerobic incubation, but no bacteria were recovered after aerobic incubation of the same samples . Of a total of eight isolates from the cementum, seven were obligate anaerobes and one was aerotolerant . The obligate anaerobes isolated were assigned to the genera Prevotella, Peptostreptococcus, Eubacterium, and Fusobacterium . The aerotolerant anaerobe was Campylobacter . From this, we conclude that bacteria can successfully invade cementum via periapical periodontal tissue, and that such bacteria may play a significant role in chronic periapical pathosis. Kansenshogaku Zasshi, 1994 Apr, 68(4), 467 - 73 {Serological study of Campylobacter jejuni infections in patients with Guillain-Barré syndrome}; Takahashi M et al.; The association between Campylobacter jejuni infection and Guillain-Barre Syndrome (GBS) was investigated serologically in 52 GBS patients . The specific antibody titers of immunoglobulin G class to C . jejuni were measured in the patient' sera using an enzyme-linked immunosorbent assay with acid-extracted antigen . The analysis showed that 31 (59.6%) of the 52 patients had evidences of recent C . jejuni infections in contrast to 16 (8.9%) of 180 healthy persons . The seropositive patients were analyzed with respect to the nature of their anticedent events . C . jejuni antibodies were found in 22 (75.9%) of the 29 patients with diarrhea, one patient with esophagus cancer, 4 (30.8%) of the 13 patients with upper respiratory infections, and 4 (50.0%) of the 8 patients without any symptoms, respectively . These findings support that there may be a close relationship between C . jejuni infection and the GBS with preceding diarrhea. Appl Environ Microbiol, 1994 Apr, 60(4), 1191 - 7 Inhibition of Campylobacter jejuni colonization in chicks by defined competitive exclusion bacteria; Schoeni JL et al.; Campylobacter enteritis in humans has been linked to consumption of chicken . Reducing Campylobacter jejuni colonization in chickens can potentially reduce Campylobacter infections in humans . In this study, the reduction of C . jejuni colonization in chicks by oral administration of defined competitive exclusion (CE) cultures, 2.5% dietary carbohydrates, or CE cultures and carbohydrates was examined . Prevention, elimination, or direct challenge of Campylobacter infection was simulated by administering treatments before, after, or at the same time as that of the Campylobacter inoculation . Additionally, the effect of maintaining high levels of protective bacteria was evaluated by administering CE cultures on days 1 and 4 (booster treatment) . All treatments reduced C . jejuni colonization . Protection by aerobically grown CE cultures was not statistically different from that by anaerobically grown CE cultures . A combination of Citrobacter diversus 22, Klebsiella pneumoniae 23, and Escherichia coli 25 (CE 3) was the most effective CE treatment . Maintaining high numbers of CE isolates by administering CE boosters did not increase protection . The greatest reduction of Campylobacter colonization was observed in schemes to prevent or eliminate C . jejuni infection . C . jejuni was not detected in the ceca of birds receiving the prevention treatment, CE 3 with mannose, representing a 62% reduction in the colonization rate . The protection factor (PF), a value combining the colonization rate and the level of infection, for CE 3 with mannose was high (> 13.2) . Fructo-oligosaccharides alone strongly prevented Campylobacter colonization . Only 8% of the chicks in this group were colonized, with a PF of > 14.3.(ABSTRACT TRUNCATED AT 250 WORDS) J Trop Pediatr, 1994 Apr, 40(2), 108 - 13 Production of DogiK: an improved Ogi (Nigerian fermented weaning food) with potentials for use in diarrhoea control; Olukoya DK et al.; As part of a programme to formulate foods to aid the control of diarrhoeal diseases, an improved ogi (the commonest weaning food in Nigeria) named DogiK has been developed . DogiK was produced by using Lactobacillus starter cultures with antimicrobial activity against diarrhoeagenic bacteria and also possessing amylolytic activity . The survival of diarrhoeagenic bacteria was investigated in locally-fermented ogi and in DogiK . The foods were inoculated with cell suspensions of Salmonella, Shigella, Campylobacter, Aeromonas, Pleisiomonas, Enteropathogenic and Enterotoxigenic Escherichia coli, Yersinia enterocolitica, and Vibrio cholerae . None of the diarrhoeagenic bacteria were detected in DogiK after 6 h whereas in the local ogi Salmonella, E . coli, and Shigella survived for 24 h or more, but showed a sharp decrease in numbers, while V . cholerae survived for 12 h . DogiK is active whether cooked or uncooked and exhibited inhibition of pathogens at neutral pH . It gives consistent quality . Preliminary investigation indicates possession of a better shelf life . Thus, DogiK may have a potential use in the prevention and treatment of diarrhoea. Oral Microbiol Immunol, 1994 Apr, 9(2), 104 - 11 Macaca fascicularis as a model in which to assess the safety and efficacy of a vaccine for periodontitis; Persson GR et al.; We have assessed Macaca fascicularis as a potential model in which to test the efficacy and safety of a vaccine for periodontitis . Twenty-eight animals were surveyed and 20 studied in more detail . Clinical periodontal status was assessed, the subgingival microflora analyzed especially for the presence and proportions of Porphyromonas gingivalis and titers and avidities of serum antibodies reactive with P . gingivalis measured . Probing depths ranged from 0.90 mm to 3.80 mm, Gingival Index scores from 0.00 to 4.00 and Plaque Index scores from 0.00 to 3.00 . About 40% of sites bled on probing . The animals manifested a subgingival flora characteristic of the anaerobic gram-negative bacteria found in human periodontal pockets, including Actinobacillus actinomycetemcomitans, P . gingivalis, Bacteroides forsythus, Campylobacter rectus, Prevotella intermedia and Fusobacterium nucleatum . P . gingivalis was detected in 70 of 80 samples studied, ranging from 0.01% to 20% of the total flora . Serum antibody reactive with antigens of P . gingivalis was observed in all animals, with titers ranging from 1.0 enzyme-linked immunosorbent assay (ELISA) unit to 25 ELISA units and avidities from 0.10 M to 2.20 M . Antibody titer and maximum percentage of P . gingivalis were inversely correlated, indicating that a humoral immune response may be effective in reducing P . gingivalis overgrowth . M . fascicularis appears to be an excellent model for use in vaccine development. Berl Munch Tierarztl Wochenschr, 1994 Apr, 107(4), 115 - 21 {Occurrence of Campylobacter and salmonellas in relation to liver changes in slaughtered poultry}; Wieliczko A; On the occasion of slaughter, 97 laying chickens, 100 broilers, 48 geese and 36 ducks were examined for Campylobacter spp . and Salmonella spp . in liver, small intestine and caeca . Pathological changes in the liver were recorded . Campylobacters and salmonellas were isolated from 61 and 18% of the laying chickens, 63 and 3% of the broilers, 54 and 15% of the geese and from 81 and 39% of the ducks, respectively . Birds with diseased liver were found more often infected with campylobacters and salmonellas than those without . This correlation could be gathered from the isolation rates from the liver and from the intestine . The occurrence of campylobacters and salmonellas together in the liver was only observed in individuals with pathological changes of the liver . The own findings suggest that besides salmonellas also campylobacters can be responsible for diseases of the liver. Berl Munch Tierarztl Wochenschr, 1994 Apr, 107(4), 109 - 15 Antigenic changes in Campylobacter spp . after adaptation to media with increased sodium chloride concentrations; Glunder G; Campylobacter strains which were adapted to growth at 3% NaCl in the medium were examined for antigenic changes . The results of the immunodot blot, the immunoprecipitation test and the immunoblot demonstrate changes in the antigenic pattern . Heat-labile and heat stable antigens of the obtained salt-tolerants variants of Campylobacter type-strains showed differences from their parental strains after cross-absorption of antisera . Differences of the antigens between salt-tolerant variants and their salt-sensitive parental strains suggest that the antigenic structure of the flagellum and possibly of outer membrane proteins has been changed by adaptation of strains to higher salt concentrations. Mol Cell Probes, 1994 Apr, 8(2), 109 - 15 Evaluation of ribotyping techniques as applied to Arcobacter, Campylobacter and Helicobacter; Kiehlbauch JA et al.; Restriction fragment length polymorphisms of ribosomal DNA (ribotyping) of many bacterial species has been useful for both epidemiologic subtyping and species identification . However, the use of ribotyping has been confined to major research and reference laboratories due to two factors: (a) the procedure must be carefully optimized for each organism one wishes to investigate and (b) most currently available protocols use hazardous chemicals or radioisotopes . The purpose of this study is to suggest an overall scheme that a clinical or research microbiologist could apply to ribotyping of any organism . In general, we recommend using a guanidium extraction method for DNA extraction, careful optimization of restriction conditions, and hybridization with non-radioactive digoxigenin-labelled probes; these procedures do not use hazardous chemicals or radioisotopes. Avian Dis, 1994 Apr-Jun, 38(2), 341 - 9 Isotype, specificity, and kinetics of systemic and mucosal antibodies to Campylobacter jejuni antigens, including flagellin, during experimental oral infections of chickens; Cawthraw S et al.; The immune response of chickens to Campylobacter jejuni infection was studied as a step in the search for vaccine candidates . One-day-old chicks orally challenged with C . jejuni strain 81116 showed significant increases in specific IgG, IgA, and IgM circulating antibodies, as detected by enzyme-linked immunosorbent assay (ELISA) . These levels peaked at 9, 5, and 7 weeks postinfection, respectively . Maternal IgG antibodies were also detected over the first 2 weeks . Specific mucosal IgG and IgA antibody levels also increased significantly . All of the birds demonstrated a major response to the 62-kDa flagellin protein by Western blotting techniques . The immunodominance of flagellin was confirmed by ELISA using an antigen preparation from an aflagellate mutant . When overlapping recombinant polypeptide fragments of flagellin were used, epitopes detected by chicken antibodies were observed in region IV, between residues 95-340 of the protein . Thus flagellin may be suitable candidate for a vaccine, although its role in protection must first be established. Kansenshogaku Zasshi, 1994 Apr, 68(4), 495 - 9 {A simple and rapid confirmation method of the bacterial contamination using polymerase chain reaction}; Kobayashi K; We studied the method for detection of bacterial contamination . Polymerase chain reaction (PCR) was used to amplify the 888 base pair of 16S ribosomal RNA (rRNA) gene fragment of various strains of bacterial species . The supernatant of bacterial suspension after treatment for 10 min at 100 degrees C was used for a template DNA . A total of 151 strains of 16 genus of Gram-negative rod and of one genus of Gram-positive cocci were confirmed and were divided into five categories by comparing digestion patterns resulting from restriction endonuclease (MluI, Eco RI and Hind III) cleavage of target rDNA fragment . These five types, such as Shigella spp . and E . coli group (Group I), other nine genera of Enterobacteria excluding Group I and Aeromonas spp . (Group II), Vibrio spp . (Group III), Campylobacter spp . and P . aeruginosa (Group IV), and S . aureus (Group V), were recognized . The group I was digested by three enzymes used, group II was by Mlu I and Eco RI but not by Hind III, group III was only by MulI, and group IV was not digested with all enzymes . The group V was sensitive to Eco RI and Hind III but was resistant to Mlu I . This method is a widely applicable technique for detection of bacterial contamination. Microbiology, 1994 Apr, 140 ( Pt 4), 847 - 55 Two types of 16S rRNA gene are found in Campylobacter helveticus: analysis, applications and characterization of the intervening sequence found in some strains; Linton D et al.; In the recently described species Campylobacter helveticus, two sizes of PCR amplicon were detected with primers homologous to conserved regions of the 16S rRNA gene . A conventionally sized gene was sequenced from the type strain, NCTC 12470, placing the new species as phylogenetically related to C . upsaliensis and the thermotolerant campylobacters . This nucleotide sequence enabled PCR primers to be designed for use in rapid molecular identification of C . helveticus and its closest phylogenetic relative, C . upsaliensis . When this assay was employed to characterize 22 'C . upsaliensis-like' isolates, twelve were identified as C . helveticus and nine as C . upsaliensis, in agreement with data obtained with a C . helveticus-specific DNA probe . A 550 bp amplicon internal to the 16S rRNA gene of C . helveticus was used to determine restriction fragment length polymorphisms (RFLPs) in genomic Southern blots, confirming that the copy number of the C . helveticus gene was three, and identifying nine 16S rRNA gene profiles . In 5/12 C . helveticus isolates identified by PCR, an enlarged amplicon was detected . The enlarged 16S rRNA gene of one of these strains, NCTC 12838, was sequenced and shown to contain an atypical intervening sequence (IVS) of 148 nucleotides . The position and size of such an IVS was inferred in the other four isolates by PCR with primers 5' and 3' to its position in NCT