Comparative analysis of Methicillin-Resistant Staphylococcus aureus (MRSA) and Borderline Oxacillin Resistant Staphylococcus aureus (BORSA) in community and food of animal origin.

The aim of this study was to illustrate the relative pervasiveness of Borderline Oxacillin Resistant Staphylococcus aureus (BORSA) and Methicillin-Resistant Staphylococcus aureus (MRSA) in community and food of animal origin and their relationship with other genetic determinants. Staphylococcus aureus isolates were subjected to E-test using the antibiotics: oxacillin, ceftriaxone, cotrimoxazole, vancomycin, genotypic tests for the genes mecA, vanA, blaZ, pvl gene and SCCmec typing. The prevalence of S. aureus (MRSA) in the food of animal origin and community settings was 21% (1.8% MRSA) and 21.9% (7.4% MRSA), respectively. SCCmec type V was prevalent among the food of animal origin, while SCCmec type IVa among the community isolates. The likelihood of MRSA presence among community isolates was three times more than in isolates from chicken and milk samples. Likewise, the likelihood of detecting pvl positive MRSA (pvl+MRSA) isolates was 4-fold higher in the community setting than in the food of animal origin. The mecA negative BORSA (mecA-BORSA) was a frequently observed phenotype among S. aureus isolates. Also, co-detection of pvl and cotrimoxazol resistance was reported in this study although there was no noteworthy correlation of cotrimoxazol resistance with the type of sample. Isolates from milk and community settings exhibit higher minimum inhibitory concentration to vancomycin (Vancomycin MIC creep, 2-4 µg/mL). SIGNIFICANCE: Current study provides the information on the statistical relationship between the genetic determinants of S. aureus with respect to sample type, and additionally the correlation that exists between the pvl and MRSA, pvl and cotrimoxazol resistance, vancomycin MIC and MRSA/Methicillin-Susceptible S. aureus (MSSA).

Prevalence, multidrug resistance and molecular typing of methicillin-resistant Staphylococcus aureus (MRSA) in retail meat from Punjab, India.

OBJECTIVES: This study reports the prevalence of methicillin-resistant Staphylococcus aureus (MRSA) in retail meat from Punjab, India. METHODS: Classical microbiological methods were applied to isolate and identify S. aureus isolates. Isolates also underwent Etest. PCR and sequencing were used to identify and characterise antimicrobial resistance genes. MLST, SCCmec and spa typing were performed. RESULTS: A total of 408 meat and 101 swab samples were processed for S. aureus isolation. Phenotypic resistance was highest to penicillin (90.97%), followed by ciprofloxacin (61.80%), tetracycline (45.14%) and erythromycin (11.11%). Isolates from chicken samples showed significantly higher MICs for tetracycline than chevon and pork samples and significantly higher MICs for trimethoprim/sulfamethoxazole and gentamicin than chevon and swab samples (P<0.05). No isolates were phenotypically resistant to vancomycin (MICs of 0.5-2µg/mL). Most isolates (52.78%, 95% CI 44.63-60.93%) were multidrug-resistant and carried resistance genes to penicillin (blaZ), oxacillin (mecA), gentamicin (aacA-aphD), erythromycin (ermB, ermC) and tetracycline (tetK, tetL, tetM). MRSA was only found in chicken samples (2.72%; 4/147). Seven S. aureus (5.07%) were borderline oxacillin-resistant (MIC range 4-8µg/mL). All MRSA were SCCmecV-pvl+-t442, among which three isolates were ST5. Their genotype was mecA+, blaZ+, aacA-aphD+, tetK+, ermC+/-. Among the erythromycin-resistant isolates, 25% were MRSA, of which 12.5% isolates expressed an inducible macrolide-lincosamide-streptogramin B (iMLSB) phenotype. CONCLUSION: These data confirm the presence of ST5-t442-MRSA-SCCmecV-pvl+ and iMLSB MRSA in meat samples, indicating a potential role of meat in the dissemination of multidrug-resistant S. aureus strains and successful MRSA lineages in Punjab.

The Prevalence of Extended-Spectrum Beta-Lactamase-Producing Multidrug-Resistant Escherichia Coli in Poultry Chickens and Variation According to Farming Practices in Punjab, India.

BACKGROUND: Agricultural use of antimicrobials in subtherapeutic concentrations is increasing in response to the rising demand for food animal products worldwide. In India, the use of antimicrobials in food animal production is unregulated. Research suggests that many clinically important antimicrobials are used indiscriminately. This is the largest study to date in India that surveys poultry production to test for antimicrobial resistance and the occurrence of extended-spectrum ß-lactamases (ESBLs) modulated by farming and managerial practices. OBJECTIVES: Our goal was to survey poultry production for resistance to eleven clinically relevant antimicrobials and phenotypic occurrence of ESBLs as modulated by farming and managerial practices. METHODS: Eighteen poultry farms from Punjab were surveyed, and 1,556 Escherichia coli isolates from 530 birds were tested for susceptibility to 11 antimicrobials using the disk diffusion method and validated using VITEK 2 (bioMérieux, Marcy-L'Étoile, France). Samples from 510 of these birds were phenotypically tested for ESBL production using the combination disk method and confirmed using VITEK 2. Generalized linear mixed models were used to infer differences in resistance profiles associated with different farming practices and facility types. RESULTS: Resistance profiles were significantly different between broiler and layer farms. Broiler farms were 2.2 [ampicillin (AMP), p=0.017] to 23 [nalidixic acid (NX), p<0.001] times more likely to harbor resistant E. coli strains than layer farms. Adjusting for farm type (broiler vs. layer), the odds of resistance (although not statistically significant) to all antimicrobials except nitrofurantoin (NIT) were higher in independent facilities (IUs) as compared to contracted facilities (CFs). Increased prevalence of multidrug resistance (MDR; 94% compared to 60% in layers), including prevalence of ESBL-producing strains (87% compared to 42% in layers), was observed in broiler farms. CONCLUSIONS: Our findings suggest that unregulated use of clinically relevant antimicrobials in Indian broiler and layer farms may contribute to the emergence of resistance and support the need to curb the nontherapeutic use of medically important antimicrobials in food animal production. https://doi.org/10.1289/EHP292.

Molecular characterization of antibiotic-resistant Staphylococcus aureus from livestock (bovine and swine).

AIM: The aim of this study was to figure the prevalence, phenotypic and genotypic antibiotic resistance (AR) pattern of Staphylococcus aureus isolated from bovine and swine nares. MATERIALS AND METHODS: Colonies with typical morphology on Baird-Parker agar supplemented with egg-yolk tellurite emulsion were selected and biochemically/genotypically identified as S. aureus. These strains were further subjected to epsilometer test for their sensitivity to various clinically important antibiotics and antibiotic susceptibility testing for amoxicillin/clavulanic acid, and double-disk diffusion testing was performed by the standard disc diffusion method following CLSI guidelines. S. aureus strains were also tested for the presence of AR genes, viz., blaZ, mecA, aacA-aphD, erm (ermA, ermB, ermC), tet (efflux genes tetK and tetL, tetM and tetO of the ribosomal protection family), and vanA. RESULTS: The nasal cavities of 17 out of 47 randomly selected bovine and 20 out of 28 randomly selected swine were positive for S. aureus, representing the prevalence of 36.2% (95% confidence interval [CI]: 22.5-49.9) and 71.4% (95% CI: 54.7-88.1), respectively. Most of the S. aureus strains showed higher resistance to penicillin (94.6%, minimal inhibitory concentration [MIC] ≥1.5 µg/ml) followed by ciprofloxacin (56.7%, MIC ≥32 µg/ml) and tetracycline (18.9%, MIC ≥32 µg/ml). About 10-15% of the strains were resistant to gentamicin (MIC 16 µg/ml) and oxacillin (MIC 6-8 µg/ml). None of the strains were resistant to vancomycin (MIC 0.25-1.5 µg/ml). In this study, 32.4% strains were resistant to three or more than three antibiotics and prevalence of this multi-drug resistant S. aureus was 45% (95%CI: 26.6-63.4) and 17.6% (95%CI: 6.7-28.5) in swine and bovine nasal samples, respectively. Four strains from pigs were borderline oxacillin-resistant S. aureus MIC 6-8 µg/ml, but none were mecA positive. Two of these strains were ß-lactamase hyperproducers. Among the resistance genes blaZ, tetK, tetL, tetM, ermB, and aacA-aphD were found. CONCLUSION: Our results demonstrated the absence of mecA and pvl gene, but the presence of multi-drug resistant S. aureus in the nares of healthy animals which has a potential to spread in a community.