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Title:	Isolation and characterization of vibrio vulnificus isolated from cockle (Anadara ganosa) and clam (Vercenaria mercenaria) from Malaysia and Qatar
Authors:	Mohammed M. Kurdi Aldulaimi (P62510)
Supervisor:	Sahilah Abd.Mutalib, Assoc. Prof. Dr.
Keywords:	Vibrio vulnificus Gram-negative bacterium Abundant Cocles Clams Dissertations, Academic -- Malaysia
Issue Date:	5-May-2016
Description:	Vibrio vulnificus is a gram-negative bacterium and abundant in the environments. It can be isolated from water, sediments, fish and shellfishes. V. vulnificus infections can cause health problems including septicemia and gastroenteritis, while wound infections are resulting from the exposure of contaminated sea water with this bacterium. The objectives of this study were to determine the presence and characterize V. vulnificus isolates from cockles (Anadara granosa) and clams (Mercenaria mercenaria) purchased from wet markets in Malaysia and Qatar, respectively. One hundred and eighty-five (n=185) shell fish samples were purchased from wet markets between July 2013 and February 2014. Ninety-eight (98) cockle samples were from Malaysia, and 87clam samples were from Qatar. A total of 60 V. vulnificus isolates were isolated, 37 isolates of which were isolated from cockles, Malaysia, and 23 isolates were isolated from Qatar. All V. vulnificus isolates were isolated from Thiosulfate-citrate-bile salts-sucrose (TCBS) and Chromagar Vibrio and identified using biochemical tests and 16S ribosomal RNA gene by polymerase chain reaction (PCR). Then, the DNA sequences were analyzed using the Base Local Alignment Search Tool (BLAST). Characterization of the V. vulnificus isolates were conducted using a phenotypic antibiogram analysis (antibiotic resistance patterns) and a genotypic analysis by Random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR), Enterobacterial repetitive interogenic consensus-PCR (ERIC-PCR) and plasmid profiling. The antibiotic resistance test was performed for 12 antibiotics by the disc diffusion method and 18 resistance patterns were observed. The isolates were highly resistant to Penicillin (93.3%), ampicillin (68.3%), clindamycin and cephalothin (66.7%) and vancomycin (63.3%), bacitracin (56.6%), erythromycin (51.7%) novobiocin (48.3%), kanamycin (5.0%) and streptomycin (1.6%). However, none of the isolates was resistant to cefoperazone and tetracycline. Plasmids were detected in 65 % of the isolates with 16 different patterns having one to four plasmids and ranging in the size from 1.5 to 35.8 kb. The results of ERIC-PCR and RAPD-PCR were analyzed using gel Compare II software. ERIC-PCR with primers ERIC1R and ERIC2 discriminated the V. vulnificus isolates into 7 clusters and 4 single isolates at a similarity level 80%. For the RAPD-PCR analysis, primer OPC6 produced 5 clusters and 3 single isolates and primer OPC11 revealed 7 clusters and 4 single isolates. Moreover, primer OPC 15 revealed 6 clusters and 3 single isolates, and primer OPAR 3 showed 6 clusters and 4 single isolates. Thus, the combination of ERIC and RAPD-PCR profiles indicated that 60 genome types showed a high degree of genetic diversity among V. vulnificus isolates isolated from different regions. The presence of V. vulnificus in the cockle and clam samples should attract attention of public health authority to monitor the cockles and clams in order to prevent food-borne infection by this pathogenic bacterium.,Certification of Master's/Doctoral Thesis" is not available
Pages:	167
Publisher:	UKM, Bangi
Appears in Collections:	Faculty of Science and Technology / Fakulti Sains dan Teknologi

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