ABSTRACT
In Germany, tuberculous lesions in slaughtered pigs due to infection with members of the Mycobacterium avium complex are increasingly reported. Contaminated food originating from pig or other livestock is discussed as potential source of human infection. M. avium isolates from man (n=45), pig (n=29), and cattle (n=13) were characterised by restriction fragment length polymorphism (RFLP) with respect to insertion sequences IS1245 and IS901 as well as by XbaI-based pulsed-field gel electrophoresis (PFGE) and the results were compared by computer cluster correlation analysis, to determine potential sources of infection in man. By PCR, 55% of animal isolates was identified as M. avium subsp. avium, and 45% as M. a. hominissuis. All human isolates belonged to M. a. hominissuis. IS1245-RFLP and PFGE resulted in two distinct main groupings reflecting the two subspecies, and dividing the isolates into several subgroups. Animal isolates of M. a. hominissuis were widely distributed within the subgroups of human isolates. M. a. avium isolates, further discriminated by IS901-RFLP, formed host-associated subgroups for animals. Comparison of RFLP patterns with those of PFGE resulted in different subgroups as well as different pairs of isolates with high similarities. Only two isolates exhibited identical patterns by both methods. In general, results of both methods support the possibility that M. a. hominissuis isolates from livestock represent a source of infection for man, probably by common environmental reservoirs. There was no evidence of human infections caused by M. a. avium in Germany.
Subject(s)
Mycobacterium avium Complex/genetics , Mycobacterium avium-intracellulare Infection/veterinary , Polymorphism, Restriction Fragment Length , Tuberculosis/veterinary , Animals , Cattle , Cattle Diseases/microbiology , Cluster Analysis , DNA, Bacterial , Disease Reservoirs/veterinary , Electrophoresis, Gel, Pulsed-Field/veterinary , Food Contamination , Food Microbiology , Genotype , Humans , Molecular Sequence Data , Mycobacterium avium Complex/classification , Mycobacterium avium Complex/isolation & purification , Mycobacterium avium-intracellulare Infection/microbiology , Polymerase Chain Reaction , Species Specificity , Swine Diseases/microbiology , Tuberculosis/microbiologyABSTRACT
Ninety-six Campylobacter upsaliensis strains that originated from Australia, Canada, and Europe (Germany) and that were isolated from humans, dogs, and cats were serotyped for their heat-stable surface antigens. All of them were genotyped by enterobacterial repetitive intergenic consensus sequence PCR (ERIC-PCR) profiling, and 83 strains were genotyped by macrorestriction analysis with the endonuclease XhoI. Eighty-four percent of the strains belonged to five different serotypes (serotypes OI, OII, OIII, OIV, and OVI), with the proportions of strains in each serotype being comparable among the groups of strains from all three continents. Two serotypes, OIII and OIV, were prevalent at rates of 35 to 40%. Serotypes OI, OII, and OVI were detected at rates of 1.5 to 15%. Between 10 and 17.7% of the strains did not react with the available antisera. Analysis of the ERIC-PCR profiles revealed two distinct genotypic clusters, which represented the German and the non-European strains, respectively. XhoI macrorestriction yielded two genotypic clusters; one of them contained 80.2% of the German strains and 34.6% of the non-European strains, and the second cluster consisted of 65.4% of the non-European strains and 19.8% of the German strains. Fourteen strains from all three continents were analyzed for their 16S rRNA gene sequences. Only two minor variations were detected in four of the strains. In conclusion, C. upsaliensis has undergone diverging processes of genome arrangement on different continents during evolution without segregating into different subspecies.
Subject(s)
Campylobacter upsaliensis/genetics , Animals , Australia , Base Sequence , Campylobacter jejuni/classification , Campylobacter jejuni/genetics , Campylobacter jejuni/isolation & purification , Campylobacter upsaliensis/classification , Campylobacter upsaliensis/isolation & purification , Canada , Cats , Consensus Sequence , DNA Primers , DNA, Ribosomal/genetics , Dogs , Genotype , Geography , Germany , Humans , Introns/genetics , Phylogeny , Polymerase Chain Reaction/methods , RNA, Bacterial/genetics , RNA, Ribosomal, 23S/genetics , Sensitivity and SpecificityABSTRACT
Campylobacter jejuni isolates of human, canine, feline, bovine and poultry origin were investigated for their genomic diversity using O-antigen typing (n = 271), SmaI (n = 158) and XhoI (n = 158) macrorestriction analysis and ERIC-PCR (n = 107). The O-antigens O:1/44, O:2, O:4 complex, O:37. O:40 were identified and 53.7% of the human and 56.1% of the animal strains were typable with the available antisera. Two ERIC-PCR pattern groups were generated representing human and animal strains as well as those exclusively of animal origin. XhoI macrorestriction analysis also distinguished 'human' and 'non-human' strain clusters, but by SmaI restriction mainly serotype-associated clusters were found. In conclusion, genomic differences may occur between 'human' and 'non-human' strains and this may reflect their potential to overcome the barrier from animals to humans.
Subject(s)
Campylobacter jejuni/genetics , Campylobacter jejuni/pathogenicity , DNA, Bacterial/analysis , O Antigens/genetics , Zoonoses , Animals , Campylobacter Infections/transmission , Cats , Cattle , DNA Primers , Dogs , Forecasting , Humans , Polymerase Chain Reaction/methods , Poultry , Restriction Mapping , Sequence AlignmentABSTRACT
A serotyping scheme based on heat-stable surface antigens was established for 101 Campylobacter upsaliensis and 10 Campylobacter helveticus strains isolated from 261 dogs and 46 cats of different ages originating from two geographically distinct regions in Germany. The prevalence of C. upsaliensis varied between 27.8% in juvenile dogs (<12 months of age) and 55.4% in adult dogs (P < 0.05). Of the cats, 19.6% harbored C. upsaliensis, whereas 21.7% carried C. helveticus. Of the C. upsaliensis isolates from both host species, 93.1% belonged to five different serogroups, two of them being prevalent at rates of 47.5 and 27.7%, with different frequencies in both regions. Six (54.6%) of the C. helveticus isolates also belonged to serotypes found among C. upsaliensis strains, whereas five (45.4%) possessed an O antigen unique for C. helveticus. In contrast, a considerable degree of genomic diversity of the isolates was assessed by macrorestriction analyses with the endonucleases SmaI and XhoI, using pulsed-field gel electrophoresis as well as enterobacterial repetitive intergenic consensus sequence PCR (ERIC PCR). Restriction with SmaI pointed towards the existence of clonal groups associated to some extent with serotypes, while restriction with XhoI disintegrated these groups to smaller noncoherent subgroups. Analysis of ERIC PCR profiles did not exhibit any associations with serotypes. In conclusion these data demonstrate the genomic heterogeneity among C. upsaliensis strains and indicate that the combination of SmaI restriction with serotyping is a useful tool to investigate the expansion of clonal groups of C. upsaliensis.
Subject(s)
Antigenic Variation , Campylobacter Infections/veterinary , Campylobacter/genetics , Genetic Variation , O Antigens/immunology , Animals , Base Sequence , Campylobacter/classification , Campylobacter/immunology , Campylobacter/isolation & purification , Campylobacter Infections/epidemiology , Campylobacter Infections/microbiology , Cat Diseases/epidemiology , Cat Diseases/microbiology , Cats , DNA, Bacterial/analysis , DNA, Bacterial/genetics , DNA, Ribosomal/analysis , DNA, Ribosomal/genetics , Dog Diseases/epidemiology , Dog Diseases/microbiology , Dogs , Electrophoresis, Gel, Pulsed-Field , Germany/epidemiology , Molecular Sequence Data , O Antigens/genetics , Polymerase Chain Reaction/methods , Polymorphism, Restriction Fragment Length , Prevalence , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA , SerotypingABSTRACT
The population of nodulating R. leguminosarum bv. viciae in soil from a grass-covered valley area which had been used for bovine slurry deposition over a period of 5 years was analyzed. For these studies, a rapid and reproducible method based on enterobacterial repetitive intergenic consensus (ERIC)-PCR was applied to identify Rhizobium strains which had infected pea nodules. Soil samples were taken from different areas and further analyzed in plant tests to determine the impact of the application of slurry (polluted or nonpolluted), the slope position (summit or toe), and exposure (north or south). After comparison of all PCR fingerprint patterns, 24 strain groups were defined. Some strain groups from the nonpolluted soil were suppressed in the polluted samples, and new strain groups were detected in the slurry-polluted soil. After analyzing relationships between the strain groups, we determined the influences of local factors on the nodulating R. leguminosarum bv. viciae population. We show that one of those local parameters, slope position, had significantly greater impact on the composition of the Rhizobium population than the presence of slurry.
ABSTRACT
A 4.6 kb DNA region of the Rhizobium meliloti strain AK631 was found to contain seven open reading frames (ORFs), all oriented in the same direction. The putative gene products of four of these ORFs were highly homologous to UreA, UreB and UreC of Klebsiella aerogenes, Proteus mirabilis, Proteus vulgaris and Canavalia ensiformis. The overall organisation of the DNA region analysed was ORF1, ureA (ORF2), ORF3, ureB (ORF4), ORF5, ORF6 and ureC (ORF7), indicating that the organisation of the urease structural genes in R. meliloti differs from that of other urease genes so far characterized. ORF1 was incomplete; only the 3' end of the coding region was present. The six complete ORFs coded for polypeptides of 11.1 (UreA), 8.9 (ORF3), 10.8 (UreB), 15.0 (ORF5), 13.8 (ORF6) and 60.7 kDa (UreC). No sequence homology to known polypeptides could be detected for the gene products of ORF1, ORF3, ORF5 and ORF6. Using a lacZ fusion and insertional mutagenesis it was shown that the seven ORFs identified were all located in the same transcription unit. For mutational analysis a resistance gene cassette was introduced into each of the complete ORFs resulting in apolar mutations. Mutations in ureA, ureB and ureC, but not in ORF3, ORF5 and ORF6, abolished urease activity in R. meliloti. The determination of hydrogen uptake in these R. meliloti mutants revealed that only ORF6 and ureB are necessary for hydrogen uptake.
Subject(s)
Genes, Plant , Sinorhizobium meliloti/genetics , Urease/genetics , Amino Acid Sequence , Base Sequence , Binding Sites , Cloning, Molecular , DNA Mutational Analysis , Hydrogenase/genetics , Molecular Sequence Data , Mutagenesis, Insertional , Open Reading FramesABSTRACT
Surface potential difference, conductance, and elasticity changes of bilayer lipid membranes induced by the antivirus drugs amantadine and remantadine were measured. An influence on the human erythrocyte shape was shown. Both drugs are stomatocytogenic. The adsorption at the cytoplasmatic membrane was electrophoretically proved. The heat-induced vesiculation is partly inhibited. No microvesicles were observed. Instead, large tails which did not detach from the cell body were seen. The general conclusion is that these amphiphilic adamantane derivatives are membrane agents which modify membrane interaction processes, possibly by influencing the bending properties.
Subject(s)
Adamantane/analogs & derivatives , Amantadine/pharmacology , Erythrocyte Deformability/drug effects , Membrane Lipids/metabolism , Rimantadine/pharmacology , Elasticity , Electron Spin Resonance Spectroscopy , Humans , Intracellular Membranes/drug effects , Time FactorsABSTRACT
EPR investigations on the vesiculation process of heated human erythrocytes were performed, using different fatty acid spin labels. Spectrin denaturation and vesiculation do not influence the fluidity of the lipid phase of the remaining membrane of human erythrocytes: Vesicles released differ in chemical composition as well as in the lipid fluidity of their membrane from the intact human erythrocyte membrane. A reduced cholesterol-to-phospholipid ratio and a depletion of spectrin was found. By changing the ionic concentration of the suspension medium an effect on membrane spectra and on vesicle release was established. The adamantane derivative amantadine causes fluidization of the human erythrocyte membrane and inhibits vesicle release. Based on these results, a model for the mechanism by which adamantane-like molecules could interact with membranes is proposed.