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Journal of Clinical Microbiology, May 2000, p. 1931-1939, Vol. 38, No. 5
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
Application of Pulsed-Field Gel Electrophoresis and
Binary Typing as Tools in Veterinary Clinical Microbiology and
Molecular Epidemiologic Analysis of Bovine and Human
Staphylococcus aureus Isolates
Ruth
Zadoks,1,2,*
Willem
van Leeuwen,3
Herman
Barkema,4
Otlis
Sampimon,5
Henri
Verbrugh,3
Ynte Hein
Schukken,1,2 and
Alex
van Belkum3
Department of Farm Animal Health, Faculty of
Veterinary Medicine, 3584 CL Utrecht,1
Department of Medical Microbiology & Infectious Diseases,
Erasmus Medical Center Rotterdam, 3015 GD
Rotterdam,3 Department of Ruminant
Health, Animal Health Service, 9200 AJ
Drachten,4 and Department of Ruminant
Health, Animal Health Service, 7400 AA
Deventer,5 The Netherlands, and Quality
Milk Promotion Services, Department of Population Medicine and
Diagnostic Sciences, College of Veterinary Medicine, Cornell
University, Ithaca, New York 14850-12632
Received 6 October 1999/Returned for modification 30 December
1999/Accepted 26 February 2000
Thirty-eight bovine mammary Staphylococcus aureus
isolates from diverse clinical, temporal, and geographical origins were genotyped by pulsed-field gel electrophoresis (PFGE) after
SmaI digestion of prokaryotic DNA and by means of binary
typing using 15 strain-specific DNA probes. Seven pulsed-field types
and four subtypes were identified, as were 16 binary types. Concordant delineation of genetic relatedness was documented by both techniques, yet based on practical and epidemiological considerations, binary typing was the preferable method. Genotypes of bovine isolates were
compared to 55 previously characterized human S. aureus
isolates through cluster analysis of binary types. Genetic clusters
containing strains of both human and bovine origin were found, but
bacterial genotypes were predominantly associated with a single host
species. Binary typing proved an excellent tool for comparison of
S. aureus strains, including methicillin-resistant S. aureus, derived from different host species and from different
databases. For 28 bovine S. aureus isolates, detailed
clinical observations in vivo were compared to strain typing results in
vitro. Associations were found between distinct genotypes and severity
of disease, suggesting strain-specific bacterial virulence.
Circumstantial evidence furthermore supports strain-specific routes of
bacterial dissemination. We conclude that PFGE and binary typing can be
successfully applied for genetic analysis of S. aureus
isolates from bovine mammary secretions. Binary typing in particular is
a robust and simple method and promises to become a powerful tool for
strain characterization, for resolution of clonal relationships of
bacteria within and between host species, and for identification of
sources and transmission routes of bovine S. aureus.
*
Corresponding author. Mailing address: Quality Milk
Promotion Services, Department of Population Medicine and Diagnostic
Sciences, Park View Technology Center I, 22 Thornwood Dr., Ithaca, NY
14850-1263. Phone: (607) 255-8202. Fax: (607) 257-8485. E-mail:
R.N.Zadoks{at}vet.uu.nl.
Journal of Clinical Microbiology, May 2000, p. 1931-1939, Vol. 38, No. 5
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
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