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Journal of dairy science v.100 no.2, 2017년, pp.1459 - 1466   SCI SCIE
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Diagnostic accuracy of a standardized scheme for identification of Streptococcus uberis in quarter milk samples: A comparison between conventional bacteriological examination, modified Rambach agar medium culturing, and 16S rRNA gene sequencing

Wald, Regina (University Clinic for Ruminants, Milk Technology and Food Science, Department of Farm Animal and Public Health in Veterinary Medicine, University of Veterinary Medicine, 1020 Vienna, Austria ); Baumgartner, Martina (University Clinic for Ruminants, Milk Technology and Food Science, Department of Farm Animal and Public Health in Veterinary Medicine, University of Veterinary Medicine, 1020 Vienna, Austria ); Urbantke, Verena (University Clinic for Ruminants, Milk Technology and Food Science, Department of Farm Animal and Public Health in Veterinary Medicine, University of Veterinary Medicine, 1020 Vienna, Austria ); Stessl, Beatrix (Institute for Milk Hygiene, Milk Technology and Food Science, Department of Farm Animal and Public Health in Veterinary Medicine, University of Veterinary Medicine, 1020 Vienna, Austria ); Wittek, Thomas (University Clinic for Ruminants, Milk Technology and Food Science, Department of Farm Animal and Public Health in Veterinary Medicine, University of Veterinary Medicine, 1020 Vienna, Austria );
  • 초록  

    ABSTRACT Bacteriological examination of milk samples is a prerequisite for pathogen-specific therapy and aids in limiting antimicrobial resistance. The aims of this study were to establish a standardized scheme for reliable Streptococcus uberis identification in routine diagnosis and to evaluate the accuracy of conventional tests and growing patterns of Strep. uberis on a selective medium (modified Rambach agar medium, MRAM) using 16S rRNA gene sequencing analysis as a reference method. We obtained isolates of presumptive Strep. uberis (n = 336) from quarter milk samples of dairy cows with intramammary infections and classified the isolates into 2 clusters using biochemical characterization. In cluster 1 (n = 280), cocci grew as non-hemolytic colonies, hydrolyzing esculin, carrying no Lancefield antigen (A/B/C/D/G) or Christie Atkins Munch-Petersen factor, and their growth was inhibited on an Enterococcus agar. Production of β- D -galactosidase on MRAM was shown by 257 of the cluster 1 isolates (91.79%), and 16S rRNA gene sequencing verified 271 (96.79%) of the isolates to be Strep. uberis. In 264 isolates (94.29%), MRAM agreed with the sequencing results. In cluster 2 (n = 56), isolates showed different characteristics: 37 (66.07%) were β- D -galactosidase-positive, and based on 16S sequencing results, 36 (64.29%) were identified correctly as Strep. uberis using biochemical methods. Identification success in this group differed significantly between routine diagnosis and MRAM application: MRAM agreed with sequencing results in 47 isolates (83.93%). To identify Strep. uberis and differentiate it from other lactic acid bacteria in routine diagnosis, we suggest using catalase reaction, hemolysis, esculin hydrolysis, and growth on enterococci agar. Isolates that show a typical biochemical profile can be identified satisfactorily with these tests. For Strep. uberis isolates with divergent patterns, application of MRAM as a follow-up test increased the diagnostic accuracy to 94.64%.


  • 주제어

    Streptococcus uberis .   mastitis .   modified Rambach agar medium.  

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