Antimicrobial Resistance Pattern and Identification of Resistant Gene in Bacteria Isolated from Meat and Milk Collected from Local Market of CMA

Abstract

Antimicrobial resistance (AMR) is known as a silent pandemic; an emerging global public health issue. The problem of AMR is similarly salient and prevalent in animals. From a public health perspective, AMR in dairy cattle, poultry meat can also jeopardize human health by the potential dissemination of AMR pathogens to humans via consumption of infected dairy products or direct contact with infected poultry and animal meat. Harshly, the dairy and poutry industry is increasingly confronted with AMR Staphylococcus aureus (S. aureus), Escherichia coli (E. coli), Campylobacter and Salmonella. This study was designed to investigate the multidrug- resistant pattern along with the most frequently isolated resistant genes of S. aureus. E. coli, Campylobacter and Salmonella from buffalo milk, buffalo meat, goat meat, beef meat, goat milk, layer meat and poultry meat in the Chittagong metropolitan area of Bangladesh. A total of 130 meat and milk samples including 40 poultry products (broiler & layer chicken meat) and 9 0 large (buffalo and cattle) and small (goat) animal products were collected from the study population. Isolation and identification of S. aureus, E. coli, Campylobacter and Salmonella were done following the standard bacteriological method. The nuc gene,16s rRNA and ST11, ST15 gene were tabbed in molecular identification of S. aureus, E. coli, Campylobacter and Salmonella respectively. In this study among 7 isolates of S. aureus four in broiler meat (20% , 95%CI 5.73 - 43.6%), one in beef meat (5%, 95%CI 0.12%- 24.8%), one in buffalo meat (10% , 95%CI 0.25%- 44.5%), one in Goat meat (5% , 95%CI 0.12%- 24.8%) were confirmed. Whereas among 47 isolates five in broiler meat (25% , 95%CI 8.65%- 49.1%), nine in beef meat (45%, 95%CI 23.05%- 68.45%), four in buffalo meat (40%, 95%CI 12.15% - 73.76%), thirteen in goat meat (65%, 95%CI 40.78%- 84.60%), six in goat milk (24% , 95%CI 9.35%- 45.12%), seven in buffalo milk (46.67% , 95%CI 21.26%-73.41%), three in layer meat (15% , 95%CI 3.2%- 37.8%) were confirmed as E. coli. Also twenty-one isolates were confirmed as Campylobacter strain out of 130 samples; Nine in layer meat (45% , 95%CI 23.05%-68.47%), twelve in broiler meat (60% , 95%CI 36.05%-80.88%). On the other hand, a total of 5 isolates were confirmed as Salmonella strain out of 130 meat and milk samples by PCR. One in beef meat (5% , 95%CI 0.12%- 24.8%), four in buffalo milk (26.67% , 95%CI 7.78%- 55.10%). xiii | P a g e xv | P a g e Among the positive S. aureus isolates, showed resistance against all tested antibiotics except Meropenem (MEM). Highest number of isolates (100%) in broiler meat was resistant against Tetracycline (TE), followed by 75% to Oxytetracycline (OT) and 50% to Ceftriaxone (CRO), Oxacillin (OX) and others. All S. aureus isolates were multi drug resistant, 57.14 % of the isolates had resistance against 4 to 6 tested antimicrobials and 42.85% had resistance to more than 7 antimicrobials. E. coli isolates were characterized with the presence of AMR genes. Afterward, the positive isolates were screened against antimicrobials using the disc diffusion technique. At a fleeting glance, alarmingly in buffalo milk, broiler meat, cattle meat and goat meat, 100% E. coli isolates of CMA showed resistance to Sulphathiazole- trimethoprim (SXT), Erythromycin (ERE), Tetracycline (TE) and Ampicillin (AMP), followed by in buffalo milk 85.71% against Ciprofloxacin (CIP), 57.14% against Gentamycin (CN), and Neomycin (N). In buffalo meat, 100% resistance showed only against Ciprofloxacin (CIP) and Ampicillin (AMP). All 47 E. coli isolates were multi drug resistant. 57.44% isolates were resistant to 4-6 antimicrobials and 40.42% were resistant to more than 7 tested drugs. In Layer meat 100% Campylobacter isolates of Chattogram area showed resistance to Erythromycin (ERE), Streptomycin (S) and Azithromycin (AZM) followed by 88.88% against Ciprofloxacin (CIP) and Sulphamethoxazole-trimethoprim (SXT), following 77.77% resistance against Cefotaxime (CTX), Ceftriaxone (CRO) and Ceftazidime (CAZ). In broiler meat, 100% Campylobacter isolates showed resistance to Erythromycin (ERE), Streptomycin (S) and Azithromycin (AZM) followed by 83.33% against Ciprofloxacin (CIP) and Sulphamethoxazole-trimethoprim (SXT). All 21 Campylobacter isolates were multi drug resistant. 23.81% isolates were resistant to 4-6 antimicrobials and 76.19% were resistant to more than 7 tested drugs. Salmonella spp. isolates of the study area from buffalo milk showed 100% resistance against Ciprofloxacin (CIP), followed by 75% against Tetracycline (TE), Sulphamethoxazole-Trimethoprim (STX), Streptomycin and 25% against Erythromycin (ERE) and Gentamycin (CN). All 5 Salmonella isolates were multi drug resistant. 40 % isolates were resistant to 2-3 antimicrobials and 60% were resistant to more than 4-6 tested drugs. Among the 7 S. aureus isolates had ESBL producing 2 blaz gene in broiler meat, cattle meat and 1 in goat meat and Erythromycin producing 1 Erm (B) gene in broiler meat and 1 in buffalo meat. Whereas, 1 Erm (C) gene in broiler meat, 1 in buffalo meat and 1 in goat xvi | P a g e meat was identified. Tetracycline producing 3 Tet (K) gene found in broiler meat and 1 in buffalo meat. Within 47 isolates of E. coli 4 Sul1 gene(sulfonamide) gene and 1 Tet (B) (tetracycline) and 1 AAC(IV) gene were identified in goat meat and 2 Sul1, 1Tet (B) and1 AAC(IV) gene were identified in buffalo milk. Also found 3 blaCMY gene, 4 Sul1, 2Tet (B), 3AAC(IV) and 1 ERE gene in cattle meat. In layer meat, only 1Sul1 and 1Tet (B) gene found. In goat milk, 2 Sul1, 3Tet (B), 2 Tet (C) and1 AAC(IV) gene were identified in E. coli isolates. Very few Campylobacter isolates of Chattogram area was identified with antimicrobial resistant gene. 3 isolates had blaTem gene both in broiler and layer meat. 2 isolates in broiler meat had Tet (C) gene. 2 Sul1 gene (sulfonamide) and 1 Tet(A)gene was identified in Salmonella spp.isolates in buffalo milk whereas 1 blaSHV,1 blaCMY and 1 Sul1 identified in cattle meat. Conclusively, the study findings will provide ample statistical evidence to develop strategies for improvement of antimicrobial stewardship, rejuvenate the antimicrobial drug channel and to develop efficacious and sustainable alternative approaches to tackling AMR crisis both in humans and livestock