Isolation and molecular identification of <i>Vibrio</i> spp. by sequencing of 16S rDNA from seafood, meat and meat products in Libya

Abstract

The genus Vibrio includes several food-borne pathogens that cause a spectrum of clinical conditions including septicemia, cholera and milder forms of gastroenteritis. Several Vibrio spp. are commonly associated with food-borne transmission including Vibrio cholerae, Vibrio parahemolyticus, and Vibrio vulnificus. Microbiological analysis for enumeration and isolation of Vibrio spp. were carried out for a total of 93 samples of seafood, meat and meat products from different geographic localities in Libya (Tripoli, Regdalin, Janzour and Tobruk). Vibrio spp. were detected by conventional cultural and molecular method using PCR and sequencing of 16S rDNA. Out of the 93 cultured samples only 48 (51.6%) yielded colonies on Thiosulfate Citrate Bile Salt agar (TCBS) with culture characteristics of Vibrio spp. More than half (n=27) of processed seafood samples (n=46) yielded colonies on TCBS, while only 44.6 % of samples of meat and meat products showed colonies on TCBS. Among cultured seafood samples, the highest bacterial count was recorded in clam with a count of 3.8 ×104 CFU\g. Chicken burger samples showed the highest bacterial count with 6.5 ×104 CFU\g. Molecular analysis of the isolates obtained in this study, showed that 11 samples out of 48 (22.9%) were Vibrio spp. Vibrio parahemolyticus was isolated from camel meat for the first time. This study is an initial step to provide a baseline for future molecular research targeting Vibrio spp. foodborne illnesses. This data will be used to provide information on the magnitude of such pathogens in Libyan seafood, meat and meat products.

MOLECULAR IDENTIFICATION AND ANTIBIOGRAM OF Enterococcus spp. ISOLATED ON ENTEROCOCCUS SELECTIVE DIFFERENTIAL (ESD) MEDIA FROM MEAT, MEAT PRODUCTS AND SEAFOOD IN LIBYA

“…# GF-GC-100, Vivantis, Malaysia) as described in previously (Azwai et al, 2016). The purified 16S rDNA amplicons underwent cycle sequencing with Big Dye® Terminator v1.1 kit (AB Applied Bioscience, TECHNE, TC-512, USA) and were sequenced on four capillary ABI PRISM® 3130-Avant Genetic Analyzer at IZSLER Istituto Zooprofilattico Sperimentale Della Lombardia e dell'Emilia Romagna, Brescia, Italy.…”

Bacteria Associated with Fresh-water Aquaculture Tilapia Fish (Oreochromisniloticus) in Suez, Egypt

“…Randomly selected samples were used for this technique either for confirmation of API results or for identification of unknown samples. The technique was performed according to Azwai1 et al [26]. DNA extraction was done using bacterial DNA preparation kit (Jena Bioscience, Thuringia, Germany).…”

Effect of technological processing on the safety of Indian mackerel (Rastrelliger kangurata) from Suez, Egypt

“…After an additional purification step using CENTRI-SEP Columns (PrincetonSeparations, Freehold, NJ), DNA sequencing was carried out by a 3500 Genetic Analyzer (Applied Biosystems, Massachusetts, USA). The obtained consensus sequences were subjected to a BLAST search through the Mega program (7.0.20) as described by Azwai1 et al(2016).…”