Biomedical subjects

Herbert Schmidt

Publications and source records attributed to Herbert Schmidt.

22 records · Page 2Linked to original sources

PubMed2000

Antibacterials that are used as growth promoters in animal husbandry can affect the release of Shiga-toxin-2-converting bacteriophages and Shiga toxin 2 from Escherichia coli strains.

Antibiotics are commonly used as growth promoters in animal husbandry worldwide. This practice has been linked to the emergence of particular antibiotic-resistant bacteria, and is now controversial. In this study, the ability of growth-promoting antibiotics to induce Shiga toxin (Stx)-converting bacteriophages from Stx-producing Escherichia coli (STEC) strains was investigated. Subinhibitory concentrations of the antibacterial growth promoters olaquindox, carbadox, tylosin and monensin were used for induction experiments. The amount of mature Stx-converting phage particles released from induced and non-induced cultures was determined, and the production of Stx was simultaneously measured by ELISA. Whereas the quinoxaline-1,4-dioxide-type antibiotics olaquindox and carbadox enhanced the release of Stx-converting phage particles from STEC cells, tylosin and monensin decreased phage induction. The production of Stx increased or decreased simultaneously with the amount of free phages. The results of this study show that particular antibacterial growth promoters can induce Stx phages. In vivo induction of Stx phages from lysogenic STEC may increase the amount of free phages in the intestine and therefore may contribute to the spread of STEC and development of new STEC pathotypes.

Animal Husbandry↗

PubMed1999

The large plasmids of Shiga-toxin-producing Escherichia coli (STEC) are highly variable genetic elements.

Shiga-toxin-producing Escherichia coli (STEC) of different serotypes are known to harbour large plasmids. The aim of this study was to investigate, using the example of the plasmid-encoded serine protease EspP, whether these plasmids are a uniform genetic element present in STEC. Examination of 201 diarrhoeagenic E. coli strains using a newly developed espP-specific PCR showed that espP is specific for STEC and present in 57% of STEC belonging to 16 different serotypes. The espP genes of the 16 STEC serotypes varied to a certain extent, as shown by nucleotide sequence and restriction enzyme analyses, but the DNA regions adjacent to the espP gene were completely different. When two further STEC-plasmid markers, the catalase-peroxidase gene katP and the enterohaemorrhagic E. coli-haemolysin gene EHEC-hlyA were included, many combinations of the three markers were found, depending in part on the serotype. In addition, strains possessing none of the three markers still harboured large plasmids. In the most prevalent STEC serogroup, O157, it was observed that the plasmid of sorbitol-fermenting STEC O157:H- lacks the espP and katP genes although both genes are present in the plasmid of the non-sorbitol-fermenting STEC O157:H7. The EHEC-hlyA gene, however, is present in both. In conclusion, this study shows that the large plasmids of STEC are not uniform genetic elements but heterogeneous in both their gene composition and arrangement.

Bacterial Toxins↗

PubMed1996

Analysis of the EHEC hly operon and its location in the physical map of the large plasmid of enterohaemorrhagic Escherichia coli O157:h7.

Almost all clinical enterohaemorrhagic Escherichia coli (EHEC) O157:H7 isolates harbour a large virulence plasmid designated pO157. In this study, pO157 of EHEC O157:H7 reference strain EDL 933 was characterized at the molecular level. A restriction map was constructed by using seven restriction enzymes, with appropriate gel electrophoretic and hybridization methods. The molecular size of pO157 was determined to be 93.6 kb. By sequencing the DNA region extending in the 3'-direction of the previously described EHEC hlyC and hlyA genes, two further genes were discovered and analysed; these were termed EHEC hlyB and EHEC hlyD. The newly discovered genes together with the EHEC hlyC and hlyA genes constitute a typical RTX (Repeats in ToXin) determinant (EHEC hly operon) with the gene order CABD. The map position of the operon was determined by hybridization experiments. Analysis of a DNA fragment carrying the downstream flanking region of the EHEC hly operon revealed an open reading frame which was highly homologous to orf1 of RepFIB, a basic replicon of IncF plasmids. It was located close to the EHEC hly operon.

Amino Acid Sequence↗

PubMed1995

The signal transducer encoded by ampG is essential for induction of chromosomal AmpC beta-lactamase in Escherichia coli by beta-lactam antibiotics and 'unspecific' inducers.

Chemical mutagenesis of the AmpC beta-lactamase-hyperinducible Escherichia coli strain SN0301/pNu305 carrying the cloned ampC and ampR genes from Citrobacter freundii OS60 gave four independent mutants in which beta-lactamase was no longer inducible, or was inducible only to a low level, by beta-lactam antibiotics. The genes ampC, ampR, ampD and ampE, which were essential for beta-lactamase induction, were functional in these mutants. In all four mutants, the sites of mutation were mapped to 9.9 min on the E. coli chromosome. Complementation with wild-type ampG restored inducibility of beta-lactamase to wild-type levels. The nucleotide sequence of all four mutant ampG alleles (ampG1, ampG3, ampG4 and ampG5) was determined. In three of the mutants, a single base exchange led to an amino acid change from glycine to aspartate at different sites in the deduced amino acid sequence. In the fourth mutant (ampG4), with low-level inducibility, the nucleotide sequence was identical to wild-type ampG. Spontaneous back-mutation of the chromosomal ampG1 mutant resulted in restoration of wild-type inducibility and a return to the wild-type ampG sequence. Unspecific induction by components of the growth medium was also dependent on intact ampG function.

Alleles↗