MOLECULAR IDENTIFICATION AND DETECTION OF EXTENDED SPECTRUM BETA LACTAMASES IN AMPICILLIN RESISTANT VIBRIO SPECIES FROM SEAFOODS IN ONDO STATE

Abstract

Vibrio species account for a significant number of foodborne infections from the consumption of raw or improperly cooked seafood and contaminated water with human feaces or sewage. The study was conducted to determine beta-lactamases production in ampicillin resistant Vibrio species isolated from seafoods collected from Ondo State coastal area. A total of one hundred and six (106) samples comprising of 10 crabs, 54 fishes, and 42 shrimps were collected from local fishermen in Erun- Amoh, Ilaje. All the samples were analyzed using standard microbiological methods. Isolated Vibrio species on Thiosulfate Citrate Bile-Salt Sucrose (TCBS) agar were subjected to antibiotic susceptibility testing by disc diffusion method. Double disc synergy test was used to determine the beta-lactamase in ampicillin resistant isolates. The genetic basis of the resistance was confirmed by plasmid profiling and plasmid curing techniques. From the results, average bacterial counts was high in shrimp with 5.0×107 cfu/g and low with 2.0×105 cfu/g. Also, average bacterial counts was high in crab with 19×107cfu/g and low with 2.0×105 cfu/g. Whereas, in fish average bacterial counts was high with 25×107 cfu/g and low with 3.0×105 cfu/g. Bacteria of the genera Bacillus, Enterobacter, Enterococcus, Escherichia, Proteus, Pseudomonas, Salmonella, Staphylococcus, and Vibrio were identified including agents of diarrhoea from seafoods. Beta-lactamases production was confirmed in two (2) V. cholerae strains that were ampicillin resistant. However, the isolates showed high degree of sensitivity to ofloxacin. Plasmids of molecular weight ranging from 3.8Kbp to 9Kbp were isolated from the three (3) Vibrio cholerae. Though, plasmid curing was successful in the third V. cholerae isolate but 9Kbp plasmids were not cured in first and second V. cholerae isolates. Post-curing sensitivity revealed that the V. cholerae strains were resistant to ampicillin and some antibiotics but, with high degree of

sensitivity to ofloxacin, levofloxacin, gentamycin cefexime, ciprofloxacin, ceftriaxone-sulbactam, cefuroxime, and augmentin. Though, most of the identified bacteria inhabit naturally in marine water environment, but contaminate seafood from human waste from faecal or urine contamination. The diversity of potential pathogens from the samples of seafoods; and the emergence of multiple antibiotic resistant ESBLs-producers Vibrio species and other pathogens is of serious Public Health concern posing a threat of poisoning when the contaminated water or seafood is consumed in quest to get nutritious food.