Characterization of staphylococci isolated from cases of mastitis and study of their R plasmids

Abstract

The emergence of drug resistant staphylococci causing mastitis deserve serious investigation. The work was intended to characterize staphylococci isolated from cases of bovine mastitis and to study their R plasmid transfer in vitro. The characterization was done using glucose and mannitol fermentation, catalase production, coagulase production, Iysostaphin sensitivity and bacteriophage typing. The antibiogram of the isolates was done by agar diffusion method using 14 chemotherapeutic agents (amoxicillin, ampicillin, bacitracin, chloramphenicol, cloxacillin, erythromycin, gentamicin, methicillin, neomycin, nitrofurantoin, penicillin, streptomycin, sulphamethoxazole and tetracycline) and by agar dilution method using eight antibiotics (ampicillin, chloramphenicol, rifampicin, streptomycin, tetracycline , gentamicin, erythromycin and penicillin) The in vitro transfer of R plasmids was tried using selected S.aureus and coagulase-negative staphylococcal isolates as donors and S.aureus RN 450RF and S.epifermidis 131S as recipients.

From 360 milk samples collected from cases of bovine mastitis 17 strains of S.aureus and 35 strains of coagulase negative staphylococci were isolated. Lysostaphin sensitivity test was positive for 90.38% of the isolates, but this could not be used to differentiate between coagulase positive and negative staphylococci. The results of phage typing revealed a predominance of group III phages over the other groups and the possible role of human strains of staphylococci in producing mastitis in animals. None of the strains were resistant to mercuric chloride. From the antibiogram rifampicin , bacitracin, neomycin , methicillin, gentamicin, cloxacillin, nitrofurantoin and chloramphenicol were found to be the drugs of choice in the treatment of bovine mastitis caused by staphylococci.

When S. aureus Rn 450RF was used as recipient six of the ten selected S.aureus isolates could transfer either one or more drug resistance markers and the mode of transfer was suspected to be by conjugation. All the four selected streptomycin resistant coagulase –negative staphylococcal donors were found to transfer the R plasmid DNA to the recipient., S.aureus RN 450RF, which was established to be through conjugation.

The drug resistant S. aureus as well as coagulase negative staphylococcal strains failed t transfer their resistance to S.epidermidis 131S.