minocycline and Mastitis--Bovine

In the present study 79 streptococcal cultures isolated from subclinical mastitis of 54 cows from seven dairy farms (A-G) in Hesse, Germany, were comparatively investigated using conventional and molecular methods. The isolates could be identified as Streptococcus agalactiae, belonging to Lancefield's serological group B by determination of cultural, biochemical and serological properties and by polymerase chain reaction (PCR)-mediated amplification of species-specific parts of the 16S ribosomal DNA, the 16S-23S rDNA intergenic spacer region and the CAMP factor gene cfb. The investigated group B streptococci were further characterized serologically for specific polysaccharide and protein antigens. Serotyping the isolates revealed a predominance of surface protein antigen X, either alone or in combination with polysaccharide antigen Ia. This could be observed for 39 isolates of farms A, B and C. Six group B streptococci from farm E displayed the serotype pattern III/Rib, two isolates from farm G showed the serotype pattern Ib/calpha. The remaining cultures from farms D and F (n=32) were non-typable. The occurrence of protein Rib could be confirmed by PCR amplification of the gene rib. The two isolates with serotype pattern Ib/calpha also reacted positively for the cbeta-encoding gene bag. Additional properties which allowed a phenotypic characterization of the S. agalactiae were the degree of pigmentation, growth properties in fluid media and soft agar, the surface hydrophobicity, the ability to hemagglutinate rabbit erythrocytes and their resistance reactions to tetracycline and minocycline. The isolates of the seven farms showed identical or almost identical characteristics. The 79 group B streptococci were additionally investigated by macrorestriction analysis of their chromosomal DNA using the restriction endonucleases SmaI, ApaI and SalI. The restriction patterns obtained by pulsed-field gel electrophoresis displayed identical or closely related patterns for the cultures of the various farms. The pheno- and genotypic characteristics of the 79 group B streptococci of the present study revealed that a single S. agalactiae strain or at least closely related subtypes of this strain were responsible for the mastitis situation of the seven farms.

Topics: Animals; Antigens, Bacterial; Bacterial Proteins; Bacterial Typing Techniques; Cattle; Chromosomes, Bacterial; Culture Media; DNA, Bacterial; Electrophoresis, Gel, Pulsed-Field; Germany; Hemagglutination; Mastitis, Bovine; Microbial Sensitivity Tests; Minocycline; Molecular Epidemiology; Polymerase Chain Reaction; Polymorphism, Restriction Fragment Length; Serotyping; Streptococcal Infections; Streptococcus agalactiae; Tetracycline

Seventy-one streptococci isolated from dairy cows with clinical mastitis were tested for tetracycline resistance. Twenty-one (30%) isolates were tetracycline resistant (Tcr), and eight hybridized with the Tet O, one hybridized with the Tet L, and one hybridized with both the Tet L and Tet K determinants. The remaining Tcr isolates did not hybridize with any of the 5 Gram-positive Tet determinants tested. The Tet O determinants were plasmid-mediated, and four selected strains transferred the Tet O determinant at frequencies of 10(-6) to 10(-8). Strains which did not hybridize with known probes were tested for resistance to minocycline. All of the Streptococcus dysgalactiae had low minimum inhibitory concentrations (MICs) for minocycline, while the S. agalactiae and the one S. uberis showed high MICs to minocycline. This suggests that at least two different uncharacterized Tet determinants exist in these isolates, one conferring high resistance to both tetracycline and minocycline and one conferring only tetracycline resistance.

Topics: Animals; Cattle; DNA Probes; DNA, Bacterial; Female; Mastitis, Bovine; Minocycline; Nucleic Acid Hybridization; Streptococcal Infections; Streptococcus; Streptococcus agalactiae; Tetracycline Resistance