Short Variable Regions flaA Gene (SVR-flaA) Diversity and Virulence Profile of Multidrug-Resistant Campylobacter from Poultry and Poultry Meat in India.

Human gastrointestinal infections caused by Campylobacter species is the second most important foodborne illness after salmonellosis worldwide. Poultry represent one of the main sources of Campylobacter organisms. In the present study, the short variable region of flagellin gene (SVR-flaA) typing was carried out to determine the variation among the circulating strains of Campylobacter jejuni and Campylobacter coli. The C. jejuni and C. coli isolated from poultry and poultry meat were screened for the presence of virulence determinants like cadF, flaA, cdtB, and wlaN gene. The screening for wlaN gene is crucial in view of the fact that most patients with Guillian Barre's (GB) syndrome with a preceding history of diarrheal illness have been found to harbor wlaN gene-positive C jejuni strains. Out of the 200 samples comprising poultry meat and cloacal swabs, 21.5% of samples were found to harbor Campylobacter spp. of which 2.5% were Campylobacter jejuni, and 19% were confirmed as Campylobacter coli. The cadF, flaA, cdtB virulence genes were detected in all the Campylobacter spp. isolated in the present study. The presence of the wlaN gene in the Campylobacter jejuni isolated in the present study may pose a public health threat with long-term human health implications. The SVR-flaA typing of twelve Campylobacter isolates obtained in the present study revealed that Campylobacter coli flaA sequence OL471375 is a new strain with a novel allele type 1,675 and peptide sequence 5 which stands deposited in pubMLST database for Campylobacter. The other flaA-SVR gene sequences identified in this study were OL471369, OL471370, OL471371, OL471372, OL471373, and OL471374. Among twelve Campylobacter spp., three distinct DdeI-RFLP patterns were observed, each varying in size from 100 to 1,000 base pairs. Antimicrobial profiling of the Campylobacter spp. isolated in the present study revealed that 50% of the strains were multidrug resistant. All the Campylobacter spp. were resistant to ciprofloxacin (CIP), ampicillin (AMP), penicillin (PEN), and nalidixic acid (NAL) whereas 57.1% of strains were resistant to tetracycline (TET) and erythromycin (ERY) 28% to amoxicillin (AMX) and enrofloxacin (ENO), 85% to amikacin (AMK). The high degree of resistance to fluoroquinolones observed in the present study is crucial in view of fluoroquinolones being drugs of choice for the treatment of human Campylobacter infections.

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.