Anti-microbial susceptibility of Streptococcus spp. isolated from bovine mastitis in Argentina.

The in vitro susceptibility to penicillin G, erythromycin and clindamycin was determined by the disc diffusion test and by E-test for a total of 47 streptococcal strains (three Streptococcus uberis, 36 Streptococcus agalactiae, eight Streptococcus dysgalactiae spp. dysgalactiae) isolated from bovine intramammary infections in Argentina. Moreover, resistance phenotypes of erythromycin-resistant streptococcal isolates was characterized. MIC90 of penicillin G, erythromycin and clindamycin for S. agalactiae were 0.75, 8.0 and 12.0 microg/ml respectively. Resistance to erythromycin and clindamycin was detected in 13 (27.6%) and 12 (25.5%) isolates respectively. No isolate was resistant to penicillin G. Resistance against macrolides, lincosamides and streptogramin B (MLS(B)) represented by the constitutive MLS(B) phenotype was present in 11 (23.4%) erythromycin-resistant isolates and two isolates (4.3%) expressed the M phenotype. The inducible MLS(B) phenotype was not identified. Results suggest that beta-lactams are the first-line antibiotics when treating streptococcal udder infections; however, the continuous monitoring of the antibiotic resistance is essential, as the emergence of resistant strains has become a growing concern on the therapy of bovine mastitis.

Antimicrobial susceptibility of coagulase-negative staphylococci isolated from bovine mastitis in Argentina.

A total of 123 isolates of coagulase-negative staphylococci isolated from bovine clinical and subclinical mastitis in Argentina from March 1998 to March 2000 was investigated for in vitro susceptibility to several antimicrobial agents. Minimum inhibitory concentrations that inhibit 90% of the isolates tested (reported in micrograms per milliliter) were: 4.40, 0.38, 4.00, 0.75, 0.75, 3.60, and 2.00 for penicillin, oxacillin, cephalothin, gentamicin, erythromycin, clindamycin, and ampicillin-sulbactam, respectively. Resistance was detected in 34 (27.6%), 4 (3.2%), 7 (5.7%), and 6 (4.8%) isolates for penicillin, oxacillin, erythromycin, and pirlimycin, respectively. No resistance was detected for gentamicin, cephalothin, or ampicillin-sulbactam. Results indicated that coagulase-negative staphylococci isolates in Argentina exhibited the highest degree of resistance to penicillin of all antimicrobial agents tested.

Molecular diversity of live-attenuated prototypic vaccine strains and clinical isolates of Staphylococcus aureus.

Bovine mastitis Staphylococcus aureus isolates and prototypic live-attenuated vaccine strains were analyzed by SmaI pulsed-field gel electrophoresis (PFGE) typing and automated ribotyping. The discriminatory index of these methods was 0.91 and 0.69, respectively. SmaI PFGE typing assigned all laboratory strains into cluster Q, which shared 49% similarity with clusters A and B, and 35% similarity with cluster C. Automated ribotyping placed laboratory strains within ribogroups different from those of bovine isolates. These methods have 70% concordance and permitted identification of the prototypic vaccine background from those of clinical isolates. This information is required before conducting field trials with the vaccine.

Genotypic analysis of Staphylococcus aureus from milk of dairy cows with mastitis in Argentina.

Staphylococcus aureus is the most prevalent pathogen causing mastitis of dairy ruminants. This study was developed to ascertain the genotypes and genealogical relationship among strains isolated from milk of bovines with mastitis in Argentina. Molecular epidemiological analysis of S. aureus was performed on 112 isolates from 21 districts. Clonality was assessed by SmaI pulsed-field gel electrophoresis (PFGE) typing, automated EcoRI ribotyping and restriction enzyme analysis of plasmid (REAP) DNA profiles. A total of 22 band patterns distributed in four clusters were found by SmaI PFGE analysis. The similarity of clusters 2, 3 and 4 with cluster 1 was 0.73, 0.69 and 0.33, respectively, and 101 of 112 isolates belonged in cluster 1. PFGE band patterns from 42 isolates within cluster I were indistinguishable from each other (type A). The second largest group of isolates with indistinguishable PFGE band patterns was subtype A11, which was composed of 19 isolates. Automated ribotyping assigned the 112 isolates into 13 ribotypes. Among these, the most prevalent ribotypes I and VI were composed of 49 and 35 isolates respectively. Although there was certain correspondence between PFGE genotypes and ribotypes, further discrimination was achieved by combining both methods. REAP DNA profile analysis was useful to provide even further discrimination between isolates with identical PFGE genotype and ribotype. The most prevalent S. aureus strains A/I and A11/VI were widely distributed in the country and were not restricted to individual nearby locations. Prevalence of these two strains varied consecutively within a period of 8 years. Whether the shift in type prevalence was due to selection of a phenotypic trait remains undisclosed.

Antimicrobial susceptibility of Staphylococcus aureus isolated from bovine mastitis in Argentina.

A total of 206 strains of Staphylococcus aureus isolated from bovine clinical and subclinical mastitis in Argentina during 1996 to 1998 were investigated for their in vitro susceptibility to several antimicrobial agents. Minimum inhibitory concentrations that inhibit 90% of the strains tested reported in micrograms per milliliters were: 1.5, 0.5, 0.75, 1.50, 0.75, 1.0 and 0.125 for penicillin, oxacillin, cephalothin, gentamicin, erythromycin, clindamycin and ampicillin-sulbactam, respectively. Resistance was detected in 83 (40.3%), 24 (11.6%), 16 (7.7%) and 7 (3.4%) S. aureus isolates for penicillin, erythromycin, pirlimycin and gentamicin, respectively. No resistance was detected for oxacillin, cephalothin and ampicillin-sulbactam. Results indicated that S. aureus isolates in Argentina exhibited high resistance to penicillin of all antimicrobial agents tested.

Capsule expression by bovine isolates of Staphylococcus aureus from Argentina: genetic and epidemiologic analyses.

Staphylococcus aureus is an important cause of bovine mastitis worldwide, and effective preventive or therapeutic modalities are lacking. Although most human S. aureus isolates produce capsular polysaccharides (CPs), few reports have described the prevalence of capsules on bovine isolates. This information is important for the rational design of a vaccine for the prevention of staphylococcal mastitis. We serotyped 195 S. aureus strains isolated between 1989 and 1997 from the milk of mastitic cows in Argentina. Only 14 (7.1%) of the strains were serotype 5, and all were recovered between 1989 and 1992. Thirteen serotype 8 strains were identified, and 12 of these were isolated between 1991 and 1994. The remaining 168 isolates were nonreactive (NR) with CP serotype 5 (CP5)- or CP8-specific antibodies. Hybridization studies performed with genomic DNA from eight NR strains revealed that only three of them carried the capsule genes. Pulsed-field gel electrophoresis (PFGE) performed with 127 of the 195 S. aureus isolates revealed that most (86%) strains belonged to one of four major PFGE groups. Although 8 of 14 CP5 isolates showed a common PFGE pattern (arbitrarily defined as A1), 31 other A1 isolates from the same time period (1989 to 1992) were not CP5 positive. In contrast, only nine PFGE type B3 isolates were recovered between 1990 and 1994, and eight of these were positive for CP8 (P < 0.0003). The results of this study underscore the variability in capsule expression by S. aureus strains isolated from different geographical regions and cast doubt on the roles of CP5 and CP8 in the pathogenesis and immunoprophylaxis of bovine mastitis in Argentina.