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. 2000 Jan;38(1):71-8.
doi: 10.1128/JCM.38.1.71-78.2000.

Characterization of Streptococcus agalactiae isolates of bovine and human origin by randomly amplified polymorphic DNA analysis

G Martinez  1 , J Harel, R Higgins, S Lacouture, D Daignault, M Gottschalk
Affiliations

Characterization of Streptococcus agalactiae isolates of bovine and human origin by randomly amplified polymorphic DNA analysis

G Martinez et al. J Clin Microbiol. 2000 Jan.

Abstract

Streptococcus agalactiae is considered one of the major causes of bovine intramammary infections. It is also found in the vaginas of women without any apparent clinical symptoms, but reports of neonatal infections, causing significant morbidity, are relatively frequent. The aim of this study was to evaluate the genetic diversity of S. agalactiae strains isolated from bovine milk and from asymptomatic women in Québec, Canada, by randomly amplified polymorphic DNA (RAPD) analysis. A total of 185 bovine isolates and 38 human isolates were first serotyped for capsular polysaccharide by double diffusion in agarose gel (bovine isolates) and coagglutination (human isolates). Strains were then studied by RAPD using 3 primers, designated OPS11, OPB17, and OPB18, which were selected from 12 primers. Thirty-eight percent of bovine isolates and 82% of human isolates could be serotyped. Prevalent serotypes were type III (28%) for bovine isolates and types V (26%) and III (24%) for human isolates. RAPD results showed that, taken together, all isolates (of bovine and human origin) shared 58% similarity. Ninety-four percent of these isolates were clustered in four groups (I, II, III, and IV) with 70% similarity among them. Three clusters, A (48 isolates), B (14 isolates), and C (32 isolates), with 79 to 80% similarity were identified within group IV, whereas the three other groups did not present any clusters. Despite some clustering of human isolates, relatively high diversity was seen among them. Relatively high heterogeneity was observed with the RAPD profiles, not only for field strains belonging to different serotypes but also for those within a given serotype.

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Figures

FIG. 1
FIG. 1
Illustration of the RAPD patterns generated with primers OPS11, OPB17, and OPB18. Lanes 1, reference strain SS615 (serotype Ia); lanes 2 nontypeable bovine isolate from region 3; lanes 3, serotype III bovine isolate from region 2; lanes 4, serotype III bovine isolate from region 7; lanes 5, serotype IV human isolate from region 4; lanes 6, serotype II bovine isolate from region 6; lanes 7, serotype V human isolate from region 4; lanes 8, serotype Ib bovine isolate from region 2; lanes M, 1-kb DNA ladder (DNA molecular size marker).
FIG. 2
FIG. 2
Genetic relationship among 223 S. agalactiae isolates (of bovine and human origins) as estimated by clustering analysis of RAPD patterns obtained with three primers. The dendrogram was generated by the unweighted pair group method with arithmetic means. H, carrier woman; B, bovine milk; UT, untypeable; AA, autoagglutinable.
FIG. 2
FIG. 2
Genetic relationship among 223 S. agalactiae isolates (of bovine and human origins) as estimated by clustering analysis of RAPD patterns obtained with three primers. The dendrogram was generated by the unweighted pair group method with arithmetic means. H, carrier woman; B, bovine milk; UT, untypeable; AA, autoagglutinable.
FIG. 3
FIG. 3
Genetic relationship among 38 S. agalactiae isolates from asymptomatic pregnant women as estimated by cluster analysis of RAPD patterns obtained with three primers. The dendrogram was generated by the unweighted pair group method with arithmetic means. UT, untypeable; AA, autoagglutinable.
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