Molecular characterization of vibro alginolyticus isolated from anadara granosa and environmental source in Kuching-samarahan distinct, Sarawak

Tay, Huey Charng (2006) Molecular characterization of vibro alginolyticus isolated from anadara granosa and environmental source in Kuching-samarahan distinct, Sarawak. [Final Year Project Report / IMRAD] (Unpublished)

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Abstract

In this study, 36 Vibrio alginolyticus strains were successfully isolated from river and cockle samples in the Kuching Samarahan district between September-November, 2005. Isolation and identification of the V. alginolyticus isolates were done by using enrichment media, selective media and standard biochemical tests. Among the 36 presumptive isolates, 15 isolates (41.67%) were randomly chosen to be analyzed using specific PCR amplification targeting the Na-translocating NADH-quinone reductase (A-Nqr) gene and generated an amplified product of 407 base pairs (bp) in size. All the isolates were further examined with respective to their antibiotic resistance, plasmid profiles and Enterobacterial Repetitive Intergenic Consensus Polymerase Chain Reaction (ERIC-PCR) analysis. Antibiotic resistance test by the Disk Diffusion method was conducted and resistance was observed towards methicillin (100.00010), penicillin G (97.22%), ampicillin (94.44%), bacitracin (88.89%), streptomycin (58.33%), gentamicin (52.78%), chloramphenicol (16.67%), tetracycline (8.33%) and cephalothin (2.78%). The multiple antibiotic resistance (MAR) index were ranging from 0.27-0.72. From the plasmid profiles analysis, 23 isolates (66.67%) were plasmid harbouring strains with plasmids sizes ranging from 1.8 MDa to more than 35.8 MDa. The ERIC-PCR analysis performed to characterize all the isolates generated fingerprint bands with molecular sizes ranging from 200 bp to 2000 bp. The genetic relationship between the isolates was analyzed using the RAPDistance package to construct a dendrogram. ERIC-PCR profiles show that the 36 isolates belonged to three major clusters comprising of 4, 17 and 19 isolates, respectively. Based on the result, ERIC-PCR has proven to be a rapid and reliable method to study the genetic relatedness of V. alginolyticus strains isolated from different location.

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