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Clinical and Diagnostic Laboratory Immunology, September 2001, p. 959-964, Vol. 8, No. 5
1071-412X/01/$04.00+0   DOI: 10.1128/CDLI.8.5.959-964.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Toxin Genes and Other Characteristics of Staphylococcus aureus Isolates from Milk of Cows with Mastitis

Ö. Akineden,1 C. Annemüller,1 A. A. Hassan,1 C. Lämmler,1,* W. Wolter,2 and M. Zschöck2

Institut für Tierärztliche Nahrungsmittelkunde, Professur für Milchwissenschaften der Justus-Liebig-Universität Gießen, 35392 Gießen,1 and Staatliches Medizinal-, Lebensmittel- und Veterinäruntersuchungsamt Mittelhessen, 35396 Gießen,2 Germany

Received 28 February 2001/Returned for modification 9 May 2001/Accepted 5 June 2001

In the present study, 103 Staphylococcus aureus strains isolated from milk samples from 60 cows with mastitis from eight different farms in seven different locations in one region of Germany were compared pheno- and genotypically and by identification of various toxins. On the basis of culture and hemolytic properties and by determination of the tube coagulase reaction, all of the isolates could be identified as S. aureus. This could be confirmed by PCR amplification of species-specific parts of the gene encoding the 23S rRNA. In addition, all of the S. aureus isolates harbored the genes encoding staphylococcal coagulase and clumping factor and the genes encoding the X region and the immunoglobulin G binding region of protein A. These four genes displayed size polymorphisms. By PCR amplification, the genes for the toxins staphylococcal enterotoxin A (SEA), SEC, SED, SEG, SEI, SEJ, and TSST-1 but not those for SEB, SEE, SEH, and the exfoliative toxins ETA and ETB could be detected. To analyze the epidemiological relationships, the isolates were subjected to DNA fingerprinting by macrorestriction analysis of their chromosomal DNAs. According to the observed gene polymorphisms, the toxin patterns, and the information given by macrorestriction analysis of the isolates by pulsed-field gel electrophoresis, a limited number of clones seemed to be responsible for the cases of bovine mastitis on the various farms.

* Corresponding author. Mailing address: Institut für Tierärztliche Nahrungsmittelkunde, Professur für Milchwissenschaften der Justus-Liebig-Universität Giebeta en, Frankfurter Str. 107, 35392 Giebeta en, Germany. Phone: 49 641 99 37663. Fax: 49 641 99 38389. E-mail: christoph.laemmler{at}vetmed.uni-giessen.de.

Clinical and Diagnostic Laboratory Immunology, September 2001, p. 959-964, Vol. 8, No. 5
1071-412X/01/$04.00+0   DOI: 10.1128/CDLI.8.5.959-964.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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