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R Helmuth

Publications and source records attributed to R Helmuth.

At least 19 recordsLinked to original sources
PubMed1995

Molecular marker analysis of Salmonella typhimurium from surface waters, humans, and animals.

Salmonella contamination of North Sea water was detected for the first time in 1988 in Germany during routine examinations of bathing areas. Since then, subsequent isolations along the coast have been reported regularly. To define the source of contamination, strains isolated from seawater and rivers were studied by molecular marker methods. Their properties were compared with those of strains originating from possible sources of contamination such as humans, cattle, and sewage treatment plant water. Plasmid profile analysis of whole bacterial populations and the determination of antibiotic resistance patterns demonstrated, that contamination through the surrounding cattle industry could be excluded. Cattle isolates belonged to a widespread clone of phage type 204c which was multiresistant and exhibited an unique plasmid pattern which was never found in sea water isolates. Outer membrane protein and lipopolysaccharide analysis failed to demonstrate differences among the Salmonella populations and proved in this case insufficient for molecular marker discrimination.

Animals↗
PubMed1994

Salmonella enteritidis phage types in Germany.

In order to monitor the epidemiological situation of S. enteritidis in Germany, in 1990-91 1138 isolates from more than 180 locations in West Germany were phage typed. 1124 strains (98.8%) from all sources were typeable, belonging to 21 different phage types (PT). PT4 strains were isolated most frequently (70.8%). In addition, PT7, 25, 34 and 8 were of epidemiological relevance with incidences of 7.2 to 4.5%. The comparison of data shows that in Germany, like in other parts of Europe, PT4 predominates. This phage type is, however, infrequent in North America, where PT8 has the highest incidence.

Animals↗
PubMed1994

Molecular typing methods for S. enteritidis.

The predominance of certain phage types of Salmonella enteritidis in various countries makes further epidemiological subgrouping necessary. Today this is achieved by using molecular typing methods. For various bacterial species, plasmid profiling, the pattern of outer membrane proteins and lipopolysaccharides, the fingerprinting of total genomic DNA including ribotyping, and multilocus enzyme electrophoretic typing, have proven very useful. When such methods have been applied to S. enteritidis, they revealed a homogeneous, clonal structure in contemporary PT4 isolates. Furthermore they indicate that the clone observed today emerged from a heterogeneous population before the onset of the epidemic.

Animals↗
PubMed1994

[Detection of verotoxin-producing E. coli in field isolates from domestic and agricultural animals in Sachsen-Anhalt].

A report is given on the detection of verotoxin-producing E. coli (VTEC) strains from field isolates of healthy or ill cattle (n = 141), pigs (n = 306), sheep (n = 15), cats (n = 29) and dogs (n = 25) in the region of the new federal land Sachsen-Anhalt. 5% of the strains isolated from cattle, 32% from pigs, 20% from sheep, 4% from dogs and 0% from cats have shown VTEC. The E. coli-strains were checked for the presence of other factors of virulence, too. A good correlation (82%) was found between the colonization factor F107 and SLT 2/2v-containing strains from pigs in the region of Sachsen-Anhalt, too. Enterohemolysin was not found in SLT 2/2v-positive strains. 91% of the VTEC, isolated from pigs, produced alpha-Hemolysin. The correlation of SLT-containing strains and the production of enterohemolysin was confirmed for ruminants, only. Plasmidprofilings of VTEC from pigs showed mainly a 60 MDa or a 68 MDa plasmid or both, too. The occurrence of heat labile (LT) and in some cases of heat stable (ST) toxin was also checked, to differentiate the VTEC-strains from the enterotoxigenic E. coli strains (ETEC). These investigations showed, that VTEC produce SLT almost without exception. Correlations and conclusions on the pathogenicity for humans are discussed.

Animals↗
PubMed1993

[Plasmid profiles of different Salmonella serovars from poultry flocks in Germany].

151 salmonella isolates belonging to 6 serotypes and originating from diverse material collected in broiler chicken farms, hatcheries and poultry slaughteries in Northern Germany during 1984 and 1990 were investigated for their plasmid DNA content. This is the first report describing plasmids of S. saint-paul, isolated from poultry lines. The different plasmids of 22 Md, 3.1 Md, 2.4 Md and 1.1 Md molecular weight can be used for further epidemiological studies of salmonellosis in poultry. The 62 Md plasmid of S. typhimurium and the 36 Md plasmid of S. enteritidis have been found to be serovar specific. The smaller plasmids (m. w. less than 5.0 Md), found in S. virchow and S. blockley can be used for further epidemiological studies.

Animals↗
PubMed1993

[Molecular biological basis for the virulence of salmonellae and the new detection methods resulting from it].

Molecular biological investigations on Salmonella isolates allowed the definition of several fundamental mechanisms of pathogenicity. They affect the confrontation of the pathogen with host defence mechanisms and colonization of the gut and other organs. Practical implications stem from the observation that especially the virulence plasmid carrying serotypes are able to cause systemic infections. Among them are the epidemiological relevant serotypes S. enteritidis and S. typhimurium. Our understanding of basic virulence mechanisms allows the development of new, faster and more sensitive diagnostic procedures. Among them are the polymerase chain reaction (PCR) and magnetic separation techniques using monoclonal antibodies coupled to superparamagnetic polystyrene beads. Presently both techniques are under development for commercial use for Salmonella detection as well.

Animals↗
PubMed1991

Heteroduplex analysis of Salmonella virulence plasmids and their prevalence in isolates of defined sources.

Strains of the Salmonella serovars S. typhimurium, S. enteritidis, S. dublin, and S. choleraesuis harbour large plasmids which are required for extraintestinal colonization after oral infection of mice. Electron microscopic heteroduplex analysis showed that these virulence plasmids share large regions of homology. Nine hundred and eighty-six isolates of different origins were analysed for the presence of these plasmids by using a cloned fragment of a S. choleraesuis virulence plasmid as a gene probe. Virulence plasmids were detected in nearly 100% of strains isolated from animal organs or human blood. Frequencies of detection ranged from 48 to 87% in strains of faecal, food or environmental origin. These results suggest that Salmonella virulence plasmids are required for systemic infections in humans and livestock.

Animals↗
PubMed1990

[Molecular biologic methods for epidemiological precision characterization of disease agents using Salmonella enteritidis from poultry as an example].

The principles of molecular biological methods for epidemiological investigation of infectious bacteria are described. They were applied for differentiating S. enteritidis isolates originating from poultry. Among the methods described are the determination of the chromosomal DNA fingerprints (BRENDA), the methods of multilocus enzyme electrophoresis, plasmid profiles, and the outer membrane protein as well as lipopolysaccharide pattern. It turned out, that S. enteritidis strains isolated before 1987, in 1988 and the ones originating from Great Britain did not differ in anyone of these criteria. Consequently one has to assume, that the strains recently causing problems originate from the population of S. enteritidis which had existed in poultry before.

Animals↗
PubMed1990

[Detection of verotoxin-producing E. coli (VTEC) in healthy cattle and swine with the DNA-DNA colony hybridization method].

With the DNA-DNA colony hybridization technique using specific gene probes for Verotoxin 1 (VT 1) and Verotoxin 2 (VT 2) 2100 E. coli strains from healthy animals were tested. Ten out of 82 milk cows (21.2%), 20 out of 212 beef cattle (9.4%) and five out of 75 pigs (6.7%) were found to carry genes for VT 1, VT 2 or both toxins, respectively. Among these strains the biotypes 5 and 6 were predominant. Some of the serotyped isolates have been described to be pathogenic for humans, like O157:H7, 082:H8, 0116, 0113, 0126 and 091, respectively. The unexpected high incidence of VTEC positive healthy animals possibly indicates a health hazard for human beings. Further investigations on the incidence of VTEC in food are necessary.

Animals↗
PubMed1990

Evolutionary genetic relationships of clones of Salmonella serovars that cause human typhoid and other enteric fevers.

Multilocus enzyme electrophoresis was employed to measure chromosomal genotypic diversity and evolutionary relationships among 761 isolates of the serovars Salmonella typhi, S. paratyphi A, S. paratyphi B, S. paratyphi C, and S. sendai, which are human-adapted agents of enteric fever, and S. miami and S. java, which are serotypically similar to S. sendai and S. paratyphi B, respectively, but cause gastroenteritis in both humans and animals. To determine the phylogenetic positions of the clones of these forms within the context of the salmonellae of subspecies I, comparative data for 22 other common serovars were utilized. Except for S. paratyphi A and S. sendai, the analysis revealed no close phylogenetic relationships among clones of different human-adapted serovars, which implies convergence in host adaptation and virulence factors. Clones of S. miami are not allied with those of S. sendai or S. paratyphi A, being, instead, closely related to strains of S. panama. Clones of S. paratyphi B and S. java belong to a large phylogenetic complex that includes clones of S. typhimurium, S. heidelberg, S. saintpaul, and S. muenchen. Most strains of S. paratyphi B belong to a globally distributed clone that is highly polymorphic in biotype, bacteriophage type, and several other characters, whereas strains of S. java represent seven diverse lineages. The flagellar monophasic forms of S. java are genotypically more similar to clones of S. typhimurium than to other clones of S. java or S. paratyphi B. Clones of S. paratyphi C are related to those of S. choleraesuis. DNA probing with a segment of the viaB region specific for the Vi capsular antigen genes indicated that the frequent failure of isolates of S. paratyphi C to express Vi antigen is almost entirely attributable to regulatory processes rather than to an absence of the structural determinant genes themselves. Two clones of S. typhisuis are related to those of S. choleraesuis and S. paratyphi C, but a third clone is not. Although the clones of S. decatur and S. choleraesuis are serologically and biochemically similar, they are genotypically very distinct. Two clones of S. typhi were distinguished, one globally distributed and another apparently confined to Africa; both clones are distantly related to those of all other serovars studied.

Alleles↗
PubMed1990

Outbreak of Salmonella typhimurium infection traced to contaminated chocolate and caused by a strain lacking the 60-megadalton virulence plasmid.

We describe an outbreak of Salmonella typhimurium infection, caused by contaminated chocolate produced by one Norwegian company, which occurred in Norway and Finland in 1987. A total of 349 bacteriologically verified cases were recorded in Norway, and 12 cases were recorded in Finland. There was a predominance of young children among the patients (median age, 6 years), many of whom developed acute hemorrhagic diarrhea. The outbreak strain exhibited a rare phage lysis pattern and a characteristic plasmid profile lacking the 60-MDa virulence-associated plasmid. DNA hybridization failed to demonstrate any DNA sequence homology between the outbreak strain and the virulence plasmid. The outbreak strain was nonlethal for orally infected mice. The finding of only less than or equal to 10 S. typhimurium cells per 100 g of chocolate in about 90% of the positive samples obtained from retail outlets suggested that an inoculum of fewer than 10 organisms may have been sufficient to cause symptomatic disease.

Cacao↗
PubMed1990

Detection and characterization of fecal verotoxin-producing Escherichia coli from healthy cattle.

Verotoxin-producing Escherichia coli isolates from feces of healthy cattle were identified by DNA hybridization with verotoxin 1- and verotoxin 2-specific gene probes. Among 259 animals investigated, 28 (10.8%) were found to carry verotoxin-producing E. coli strains. Characterization of the verotoxin-producing isolates revealed a heterogeneous population in terms of serotype and toxin type. Nearly 40% of the strains belonged to serogroups known to be pathogenic for humans, i.e., O22, O39, O82, O91, O113, O116, O126, and O136. Two isolates from different bulls were identified as serotype O157:H7. Results obtained in this study indicate that cattle may be an important source of verotoxigenic E. coli involved in human disease.

Animals↗
PubMed1990

HLA-DQ alpha allele and genotype frequencies in various human populations, determined by using enzymatic amplification and oligonucleotide probes.

Allele and genotype frequencies at the HLA-DQ alpha locus have been determined by the use of polymerase chain reaction (PCR) amplification and nonradioactive oligonucleotide probes. The probes define six alleles and 21 genotypes in a dot-blot format. A total of over 1,400 individuals from 11 populations has been typed by two different laboratories using this method. In contrast to some variable-number-of-tandem-repeat markers that have been used for identity determination, DQ alpha genotype frequencies do not deviate significantly from Hardy-Weinberg equilibrium in all populations studied. The distribution of alleles varies significantly between most of these populations. In Caucasians, the allele frequencies range from 4.3% to 28.5%. In this population, the power of discrimination is .94, and, for paternity determination, the power of exclusion is .642. These population data will allow the use of the HLA-DQ alpha marker in paternity determination, the analysis of individual identity in forensic samples, and anthropological studies.

Alleles↗
PubMed1988

Toward a population genetic analysis of Salmonella: genetic diversity and relationships among strains of serotypes S. choleraesuis, S. derby, S. dublin, S. enteritidis, S. heidelberg, S. infantis, S. newport, and S. typhimurium.

Variation in the chromosomal genomes of 1527 isolates of eight common serotypes (O and H antigen profiles) of Salmonella was assessed by analysis of electrophoretically demonstrable allelic polymorphism at 23 metabolic enzyme loci. Seventy-one distinctive electrophoretic types, representing multilocus genotypes, were identified. A basically clonal population structure was indicated by the presence of strong linkage disequilibrium among enzyme loci, the association of each serotype with a relatively small number of multilocus enzyme genotypes, and the global distribution of certain genotypes. For each of six of the serotypes, 83-96% of isolates were members of a single clone. The occurrence of each of four serotypes (S. derby, S. enteritidis, S. infantis, and S. newport) in isolates of clones belonging to several evolutionary lineages, some of which are distantly related, suggests that the horizontal transfer and recombination of chromosomal genes mediating expression of cell-surface antigens has been a significant process in the evolution of the salmonellae. Two divergent clone clusters of S. derby differ in the relative frequency with which they cause disease in birds versus mammals, and two major lineages of S. newport differ in the frequency with which their clones are associated with disease in humans versus animals.

Alleles↗
PubMed1986

Epidemiology and chromosomal location of genes encoding multiresistance in Salmonella dublin.

Multiresistant Salmonella dublin strains isolated between 1971 to 1984 were investigated for their genetic and physical properties. It turned out that the multiresistance of the 1971 to 1973 isolates was plasmid-encoded. In the following years, however, the plasmid profiles of the strains were indistinguishable from sensitive isolates and the genes encoding the multiresistance were located on the Salmonella chromosome.

Chromosome Mapping↗
PubMed1985

Epidemiology of virulence-associated plasmids and outer membrane protein patterns within seven common Salmonella serotypes.

Antibiotic-sensitive Salmonella isolates belonging to seven common serotypes and originating from 29 different countries from all continents were investigated for their plasmid DNA content (337 isolates) and their outer membrane protein profiles (216 isolates). Of the S. typhimurium, S. enteritidis, S. dublin, and S. choleraesuis isolates, 90% or more carried a serotype-specific plasmid. The molecular sizes of the plasmids were 60 megadaltons (Md) for S. typhimurium, 37 Md for S. enteritidis, 56 Md for S. dublin, and 30 Md for S. choleraesuis. The outer membrane protein profiles were homogeneous within each of the seven serotypes, except that a minority of S. enteritidis and S. dublin strains were lacking one major outer membrane protein. Virulence studies were performed with 39 representative strains by measuring the 50% lethal doses (LD50S) after oral infection of mice. The LD50 values obtained for plasmid-positive strains of S. typhimurium, S. enteritidis, and S. dublin were up to 10(6)-fold lower than the values obtained for the plasmid-free strains of the same serotype. Only the plasmid-positive strains could invade the livers of orally infected mice, and only they were resistant to the bactericidal activity of 90% guinea pig serum. Strains of S. infantis were generally plasmid free, whereas S. panama and S. heidelberg isolates carried heterogeneous plasmid populations. The virulence properties of the latter three serotypes could not be correlated with the predominant plasmids found in these strains.

Bacterial Outer Membrane Proteins↗