Evaluation of tRNA intergenic spacer length polymorphism analysis as a molecular method for species identification of streptococcal isolates from bovine mastitis

Sixty-nine bovine mastitis streptococci belonging to the species Str. agalactiae (n=13), Str. dysgalactiae (n=16), Str. canis (n=22), Str. uberis (n=20) and Str. parauberis (n=4) and six reference strains of the five streptococcal species were examined for their tRNA gene intergenic length polymorphism (tDNA-ILP) fingerprint pattern. Epidemiologically unrelated isolates from bovine mastitis cases were selected by macrorestriction analysis with pulsed-field gel electrophoresis (PFGE). Their results were compared with those obtained from biochemical and serological studies and with those obtained by PCR-mediated identification amplifying speciesspecific gene segments of the five streptococcal species. According to the present results tDNA-ILP allowed a correct identification of all Str. agalactiae, Str. uberis and Str. parauberis strains investigated also including the reference strains of each species showing species-specific banding pattern. However, all Str. dysgalactiae ssp. dysgalactiae and all Str. canis strains appeared with an undistinguishable pattern which did not allow an identification of the species.