A survey for sarcosporidial cysts in east African game animals.
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Biomedical subjects
Publications and source records attributed to D Protz.
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We investigated 158 samples of shortly ripened raw sausages bought in supermarkets of Dessau within 4 month. In 14 (8.8%) samples Verotoxin-producing E. coli were detected. 13 VT-positive samples were found in the group of easily spread raw sausages. The 14 isolates belonged to 6 different O-serotypes. 4 VT1-, 8 VT2- and 2 VT1/VT2-producers were found. 4 isolates belonged to serogroups which were already described in WHO tables and associated with EHEC infections in human beings. One strain of serogroup O22: H8, isolated from a "Teewurst", possessed the complete virulence gene combination of EHEC (eae, hlyA, stx). The detection procedure, already successfully used for detection and isolation of VTEC from raw milk, soft cheese and raw minced beef showed a sensitivity of approximately 10 CfU/25 g of raw sausages. It has to be considered that VTEC are frequently (8.8%) present in shortly ripened raw sausages. The group of easily spread raw sausages has a higher VTEC-contamination rate than firm raw sausages. Raw sausages, especially easy to spread types, belong to the risk foods for EHEC-infections in human beings.
Enterotoxigenic Escherichia coli (ETEC) may produce heat-labile (LT) and heat-stable (STI or STII) enterotoxins. Differentiation between ETEC and other pathogenic and non-pathogenic E. coli as well as other Gram-negative bacteria responsible for induction of diarrhoea, requires isolation, biochemical identification and determination of toxins (or their genes--elt, estI, estII). A multiplex polymerase chain reaction (PCR) system for the rapid and specific detection of enterotoxin-gene-positive E. coli was developed. The primers described by other authors, specific for the universal stress protein A (UspA) of E. coli and enterotoxin genes were used and allowed a simultaneous amplification of the E. coli-specific uspA and the respective toxin genes. The specificity of this multiplex PCR system was confirmed by testing ETEC, non-ETEC and other non-E. coli bacteria. The specific 884 bp uspA gene and 280 bp (eltI), 166 bp (estI) or 278 bp (estII) amplification products were generated with the respective ETEC strains whereas no amplification was detected with non-E. coli bacteria. The multiplex PCR developed allowed the rapid and specific identification of enterotoxin-producing E. coli colonies directly grown from faecal samples of pigs with diarrhoea. The test may be used as a method for the determination of ETEC among other pathogenic groups of E. coli and other Gram-negative enteric isolates.
Methods for making check ups of STEC-Screening-PCRs by using internal and external systems are given. A control-DNA with identical primer binding sites in relation to the target-DNA (stx-genes) was produced and by HPLC-chromatography quantified. It was used in an internal system. But we had to use too high concentrations of this DNA for making the check up in each sample. A false negative result could be shown in samples with a low stx content as a result of this coamplification process. Therefore we used an external system. A control-DNA with heterologous primer binding sites in relation to the stx-screening-PCR and the special primers were given in each vessel. The optimized system was used for making investigations of some faecal samples from cattle.
In the last years the number of isolations of Salmonella enterica subspecies enterica serovar paratyphi B (S. paratyphi B) sent to the national salmonella reference laboratory of Germany has increased steadily. Most of the isolates originated from fowl or poultry products. The bacteriological, serological and biochemical properties of the isolates were investigated. Special emphasis was given to the utilization of d-tartrate which subgroups the serovar. All of them belonged to the d-tartrate positive variant, which is generally considered less virulent for humans and was formerly called S. java. The performance of various tests is compared and in addition the possibility of the spread within the production line is discussed.
A method for specific isolation of VT(+)-strains in raw milk is given. DNA-hybridization technique with DIG-labeled PCR-amplificates as probes are the basis. No background is seen by using "DIG Easy Hyb" solution and nylon membranes for colony- and plaque-hybridization (Boehringer Mannheim GmbH). Marked colonies are visible on the membranes after detection. So it is possible to select these colonies from a masterplate. The results are available within one day (without enrichment and membrane preparation). After stripping the membranes can be used for a new hybridisation to detect another factor of virulence.