PubMed Health⌕ Search

Biomedical subjects

W Rabsch

Publications and source records attributed to W Rabsch.

At least 37 records · Page 2Linked to original sources

Salmonella typhimurium IroN and FepA proteins mediate uptake of enterobactin but differ in their specificity for other siderophores.

Salmonella typhimurium possesses two outer membrane receptor proteins, IroN and FepA, which have been implicated in the uptake of enterobactin. To determine whether both receptors have identical substrate specificities, fepA and iroN mutants and a double mutant were characterized. While both receptors transported enterobactin, the uptake of corynebactin and myxochelin C was selectively mediated by IroN and FepA, respectively.

Bacterial Outer Membrane Proteins↗

Molecular characterization of Salmonella enterica subsp. enterica serovar Typhimurium DT009 isolates: differentiation of the live vaccine strain Zoosaloral from field isolates.

A total of 25 epidemiologically unrelated Salmonella enterica subsp. enterica (S.) serovar Typhimurium DT009 isolates from various human and animal sources, the original S. Typhimurium DT009 Zoosaloral live vaccine strain and two Zoosaloral strains reisolated from vaccinated chickens were investigated by various molecular typing methods (I) to determine the most suitable method or combination of methods for the differentiation of DT009 field isolates and (II) to investigate which molecular methods are suitable to differentiate the Zoosaloral live vaccine strain from field isolates of the same phage type. Based on the results of plasmid profile analysis, IS200 typing and macrorestriction analysis with XbaI, SpeI and BlnI, the 28 S. Typhimurium DT009 isolates were assigned to 16 different genomic groups, one of which was exclusively represented by the original and the reisolated Zoosaloral strains. IS200 typing was the most discriminatory single method for the differentiation of the DT009 isolates followed by plasmid profile analysis and BlnI-macrorestriction analysis. The Zoosaloral vaccine strain differed from the DT009 field isolates by its unique HindIII-fragment pattern of the virulence plasmid and by its unique SpeI-macrorestriction pattern.

Animals↗

A comparative study on the frequency of prophages among natural isolates of Salmonella and Escherichia coli with emphasis on generalized transducers.

Several collections of natural isolates of the genus Salmonella and of the species Escherichia coli were studied for the release of viable temperate phages. The results indicated that functional prophage genomes may be a common constituent of all bacterial genomes of the investigated strains. About 99% of the Salmonella phages are capable of generalized transduction of chromosomal host markers and plasmids. The ratio of transducing E. coli phages is significantly lower.

Animals↗

IroN, a novel outer membrane siderophore receptor characteristic of Salmonella enterica.

Speciation in enterobacteria involved horizontal gene transfer. Therefore, analysis of genes acquired by horizontal transfer that are present in one species but not its close relatives is expected to give insights into how new bacterial species were formed. In this study we characterize iroN, a gene located downstream of the iroBC operon in the iroA locus of Salmonella enterica serotype Typhi. Like iroBC, the iroN gene is present in all phylogenetic lineages of S. enterica but is absent from closely related species such as Salmonella bongori or Escherichia coli. Comparison of the deduced amino acid sequence of iroN with other proteins suggested that this gene encodes an outer membrane siderophore receptor protein. Mutational analysis in S. enterica and expression in E. coli identified a 78-kDa outer membrane protein as the iroN gene product. When introduced into an E. coli fepA cir fiu aroB mutant on a cosmid, iroN mediated utilization of structurally related catecholate siderophores, including N-(2,3-dihydroxybenzoyl)-L-serine, myxochelin A, benzaldehyde-2,3-dihydroxybenzhydrazone, 2-N,6-N-bis(2,3-dihydroxybenzoyl)-L-lysine, 2-N,6-N-bis(2,3-dihydroxybenzoyl)-L-lysine amide, and enterochelin. These results suggest that the iroA locus functions in iron acquisition in S. enterica.

Amino Acid Sequence↗

Studies of the presence of the virulence factors, adhesion, invasion, intracellular multiplication and toxin formation in salmonellas of different origin.

Salmonellas of different origin were classified into two groups (11 strains of common serovars which had been isolated from organs of calves having died from salmonellosis and 18 strains belonging to rare serovars which showed uncommon metabolic characteristics and had been isolated from spices and spiced foods). The strains were examined with regard to different virulence parameters. All salmonellas investigated possessed the genetic information on invasion (invA) and toxin formation (stn). They adhered equally well to epithelial cells, could penetrate into these and survive and multiply inside the cells. The formation of toxic substances could be detected in all strains after co-cultivation with epithelial cells in the CHO-K1 test. Significant differences between the groups of strains could be demonstrated only for the invasion of epithelial cell monolayers. Since adhesion, invasion and the ability of intracellular survival and multiplication as well as toxin formation constitute virulence parameters of salmonellas, it must be assumed that also the Salmonella serotypes studied which have been rarely observed epidemiologically constitute a risk for humans.

Animals↗

Iron-regulated excretion of alpha-keto acids by Salmonella typhimurium.

Excretion of alpha-keto acids by clinical isolates and laboratory strains of Salmonella typhimurium was determined by high-performance liquid chromatography analysis of culture supernatants. The levels of excretion increased markedly with increasing iron stress imposed by the presence of alpha,alpha'-dipyridyl or conalbumin in the medium. The major product was pyruvic acid, but significant concentrations of alpha-ketoglutaric acid, alpha-ketoisovaleric acid, and alpha-ketoisocaproic acid were also observed. Maximal excretion occurred at iron stress levels that initially inhibited bacterial growth; the concentration of alpha,alpha'-dipyridyl at which this was observed differed between strains depending on their ability to secrete and utilize siderophores, suggesting that the intracellular iron status was important in determining alpha-keto acid excretion. However, prolonged incubation of the siderophore-deficient S. typhimurium strain enb-7 under conditions of high iron stress resulted in significant delayed bacterial growth, promoted by tonB-dependent uptake of iron complexed with the high accumulated levels of pyruvic acid and other alpha-keto acids. Strain RB181, a fur derivative of enb-7, excreted massive amounts of alpha-keto acids into the culture medium even in the absence of any iron chelators (the concentration of pyruvic acid, for example, was >25 mM). Moreover, RB181 was able to grow and excrete alpha-keto acids in the presence of alpha,alpha'-dipyridyl at concentrations threefold greater than that which inhibited the growth of enb-7.

2,2'-Dipyridyl↗

TonB-dependent iron supply in Salmonella by alpha-ketoacids and alpha-hydroxyacids.

A range of structurally diverse alpha-ketoacids and alpha-hydroxyacids promoted the growth of isolates of Salmonella serovars S. typhimurium, S. enteritidis, S. agona, S. paratyphi and S. stanleyville in iron restricted conditions in the absence of functional siderophores. Growth stimulation, observed both in cross-feeding tests on solid medium and in liquid cultures, and uptake of 55Fe in the presence of alpha-ketoisocaproic acid, were TonB dependent in S. typhimurium. In this respect the mechanism is distinct from a previously described Serratia marcescens system (sfuABC); the presence of the cloned sfuABC genes mediated tonB-independent uptake by S. typhimurium of iron complexed with alpha-ketoacids.

Bacterial Proteins↗

Ferrioxamine E-supplemented pre-enrichment and enrichment media improve various isolation methods for Salmonella.

Supplementation of pre-enrichment broth and enrichment broth media with ferrioxamine E (1 microgram/ml) significantly improved the recovery of Salmonella from artificially or naturally contaminated foods. Based on the selectivity of ferrioxamine E, Salmonella enteritidis and S. typhimurium could be isolated also from various mixed cultures (one Salmonella cell in 10(3)-10(4)-fold concentration of cells of competitors) by shaking for 6 h in supplemented buffered peptone water followed by cultivation on XLD- or XLT-4 agars. Isolation of Salmonella from these pre-enrichment cultures by use of Dynabeads-Anti-Salmonella was highly effective. 27 S. typhimurium strains were isolated from 762 naturally infected chicken giblets by use of unsupplemented Tetrathionate broth. However, 33 S. typhimurium isolates were obtained with ferrioxamine E-supplemented Tetrathionate broth from the same samples. Three Salmonella isolates out of 50 evenly divided meat meal samples were obtained by use of ferrioxamine E-supplemented buffered peptone water followed by direct streaking onto XLD- and Rambach agars, no Salmonella isolates could be detected by the conventional method.

Animals↗

Identification of DNA gyrase A mutations in ciprofloxacin-resistant isolates of Salmonella typhimurium from men and cattle in Germany.

Six multiply resistant isolates of Salmonella typhimurium var. copenhagen with high-level resistance to fluoroquinolones (e.g., MIC of ciprofloxacin: 32 micrograms/ml) were isolated from human patients (n = 3) and from cattle (n = 3). The isolates were examined by complementation tests using a set of broad-host-range plasmids, which carry either the gyrA+ or the gyrB+ genes or a combination of both from Escherichia coli K-12. The results indicated a combination of gyrA and gyrB mutations in all isolates. Subsequent direct sequencing of PCR-generated internal DNA fragments of gyrA revealed an identical double mutation in all six isolates (Ser-83-->Ala and Asp-87-->Asn). In addition, the results of phenotypic (i.e., phagetype, biotype, serotype) and genotypic characterization [i.e., ribotyping and polymerase chain reaction fingerprinting (PCR-fingerprinting)] were identical for all six isolates and were distinguishable from a quinolone-susceptible strain of the same serovar and an unrelated isolate of S. typhimurium. These data indicate the clonal identity of the fluoroquinolone-resistant strains of S. typhimurium isolated from men and cattle in Germany.

Adolescent↗

tRNA genes and pathogenicity islands: influence on virulence and metabolic properties of uropathogenic Escherichia coli.

The uropathogenic Escherichia coli strain 536 (O6:K15:H31) carries two unstable DNA regions on its chromosome which were termed pathogenicity islands (Pais). Both pathogenicity islands, Pai I and Pai II, are incorporated into tRNA specific loci: Pai I is located in the tRNA gene for selenocysteine (selC), and Pai II is integrated in the leucine-specific tRNA locus leuX. Mutant strain 536-21 has lost the two pathogenicity islands together with the intact tRNA genes. While 536 is a virulent strain, 536-21 has lost a number of properties, including in vivo virulence. In previous publications we reported that the genes coding for two haemolysins (hly I, hly II) and P-related fimbria (prf) are located on the Pais. In this paper, we demonstrate that the expression of other gene products influencing metabolic properties in addition to in vivo virulence are strongly dependent on the intact tRNA loci selC and leuX. In order to determine the influence of the two tRNAs on the expression of these properties, the genes selC and leuX were cloned from the genome of strain 536 and then introduced into the mutant 536-21. Our results clearly show that the seleno-cysteine-specific tRNA (tRNA(Sec)) directly influences the ability of the bacteria to grow under anaerobic conditions, because selenocysteine is part of the enzyme formate dehydrogenase (FDH) which is involved in mixed acid fermentation. The rare leucine-specific tRNA5(Leu), encoded by leuX, influences a number if properties including type 1 fimbria production, flagellation and motility, production of enterobactin and serum resistance, and is also necessary for full in vivo virulence. While the tRNA(Sec) is directly involved in the production of FDHs, the leuX specific tRNA5(Leu) appears to influence the expression of various factors through specific transcriptional or translational control mechanisms.

Anaerobiosis↗

Iron supplying systems of Salmonella in diagnostics, epidemiology and infection.

Well-known and newly characterised mechanisms, both endogenous and exogenous, for the uptake of iron by Salmonella are outlined, and their possible roles at various stages in infection are discussed. The contributions of a detailed understanding of iron supplying systems to techniques for diagnosis, epidemiology and disease management are described.

Bacterial Outer Membrane Proteins↗

[Epidemiological and microbiological studies on salmonellosis in Russia].

The occurrence of the most important Salmonella serovars in Russia from 1980 to 1993 is reported. Selected Salmonella strains isolated in Russia from 1951 to 1993 were characterized by serotype, phage type, resistance pattern, plasmid profile and detection of aerobactin production. The incidence of salmonellosis was different in the various administrative districts and varied between 0-235 cases/100,000 inhabitants. Since the middle of the eighties both the overall increase of salmonellosis and the replacement of S. typhimurium by S. enteritidis was also demonstrable in Russia. Nosocomial infections due to multiply resistant S. infantis strains and S. typhimurium strains which caused severe septicaemic courses in children were a special epidemic problem. These infections mainly occurred via faecal-oral transmission. In contrast ot Western Europe most of the S. enteritidis strains isolated in Russia belonged to phage type 1/1. Eight phage types were determined additionally. The S. typhimurium strains isolated in nosocomial infections belonged to a definite phage type and carried a 95 MDa plasmid. This plasmid was also detected in the nosocomial S. infantis strains, S. haifa strains, and S. panama strains. All these nosocomial strains formed aerobactin.

Anti-Bacterial Agents↗

Increased occurrence of KCN-resistant Salmonella strains.

104 KCN-resistant Salmonella strains were detected among 1391 strains typed (7.4%) during the first 7 months in 1993. In particular Salmonella enterica strains of subspecies I isolated from humans and foods exhibited this character. Those KCN-resistant Salmonella strains did not belong to a special antigen formula. The second flagellar phase could not be detected in most of these strains. KCN resistance does not seem to be plasmid-encoded.

Drug Resistance↗

Desferrioxamine-promoted virulence of Yersinia enterocolitica in mice depends on both desferrioxamine type and mouse strain.

The effects of desferrioxamine B (DFOB) and G (DFOG) on growth promotion and virulence of Yersinia enterocolitica as well as on T cell activation and proliferation were investigated. Both desferrioxamines promoted growth of Y. enterocolitica O:8 (WA-314) and O:3 (Y-108). DFOB had a greater immunosuppressive effect on T cells than did DFOG. These results suggest a dual role of DFOB in yersiniosis: growth promotion of the pathogen and immunosuppression of the host. The LD50 of both Yersinia strains for mice was reduced by DFOB but not by DFOG. However, the LD50 of yersiniae was reduced by DFOB to a greater extent in Yersinia-resistant C57BL/6 than in Yersinia-susceptible BALB/c mice. The different impact of DFOB on the LD50 of Y. enterocolitica in C57BL/6 and BALB/c mice might be due to an immunomodulating effect of DFOB.

Animals↗

Evaluation of Rambach agar for detection of Salmonella subspecies I to VI.

Salmonella strains belonging to subspecies I to VI were investigated for colony color when grown on Rambach agar. Most strains of Salmonella subspecies I, II, IV, and VI behaved as described. All strains of Salmonella subspecies IIIa, IIIb, and V produced beta-D-galactosidase and blue-green colonies which could not be distinguished in color from Escherichia coli and other lactose-fermenting members of the family Enterobacteriaceae.

Agar↗

[Iron availability in Salmonella in diagnosis, epidemiology, and the infectious process].

Well known and new iron supplying systems were described. Salmonella use siderophores of different chemical structures and uptake-systems for self-produced siderophores and siderophores produced by other microorganisms. Iron supplying systems contribute to new methods in diagnosis, epidemiology, vaccination, and development of new siderophore-beta-lactams. It is hoped to obtain new insights in the pathogenesis of salmonella by using methods of iron supplying systems.

Animals↗

[Current epidemiological status of salmonellosis of humans in Germany].

Infections caused by Salmonella typhi and Salmonella paratyphi B have special epidemiological significance. However, the enteritis caused by Salmonella enteritidis and Salmonella typhimurium dominates. A complex typing of these serovars is the prerequisite for both, an effective surveillance of salmonellosis and the detection of occurring replacements and variations of the pathogens. The increasing occurrence of Salmonella in spices since 1993 is epidemiologically remarkable. In this connection, serotypes such as Salmonella javiana, S. rubislaw, and monophasic, KCN-positive serovars dominate. The transmission route man-man is of no special epidemiological importance at present in Germany.

Enteritis↗