Discovery on Antibiotic Resistance Patterns of Vibrio parahaemolyticus in Selangor Reveals Carbapenemase Producing Vibrio parahaemolyticus in Marine and Freshwater Fish.

Sunny Hei Wong,Vengadesh Letchumanan,Jodi Woan-Fei Law,Learn-Han Lee,Nurul-Syakima Ab Mutalib

doi:10.3389/fmicb.2018.02513

Abstract

Vibrio parahaemolyticus, a Gram-negative halophilic bacterium is often associated with fish and fishery products, thus causing gastroenteritis in humans upon ingestion of contaminated food. V. parahaemolyticus has become a globally well-known pathogen with yearly reported cases in many countries. This study aimed to discover the antibiotic resistance patterns of V. parahaemolyticus as well as detect Carbapenem resistant isolates from marine and freshwater fish in Selangor. A total of 240 freshwater and marine fish samples collected from wet market and supermarket in Selangor were tested for the presence of V. parahaemolyticus. All the fish samples were determined positive for V. parahaemolyticus using conventional microbiological culture-based method. The toxR gene were detected via polymerase chain reaction (PCR) in 165/240 (69%) isolates. The two-virulence factor of V. parahaemolyticus, thermostable direct hemolysin (tdh) and TDH-related hemolysin (trh) was screened via PCR. As such, four isolates were trh+and none were tdh+. Majority of the isolates presented high resistance to ampicillin (88%), amikacin (64%), and kanamycin (50%). In addition, this study identified 19-imipenem resistant isolates isolated from freshwater and marine fish samples. Further analysis of these 19-imipenem resistant isolates revealed that the resistance toward imipenem was plasmid mediated after plasmid curing assay. The multiple antibiotics resistance index was >0.2 for 70% of the isolates. In summary, the results confirm the presence of V. parahaemolyticus in freshwater and marine fish samples in Selangor, Malaysia. To our best knowledge, this is the first report discovering the antibiotic resistant patterns and Carbapenem-resistant isolates of V. parahaemolyticus isolated from marine and freshwater fish samples in Selangor.

Highlights

Vibrionaceae family within the class of Gammaproteobacteria comprises of Gram-negative halophilic bacteria, straight or curved rods, ubiquitous and indigenous in aquatic environments (Tison and Kelly, 1984; Tantillo et al, 2004; Sawabe et al, 2013)
Vibrio parahaemolyticus is among the member of this genus that been regarded as important human pathogenic bacteria (Su and Liu, 2007; Iwamoto et al, 2010; Bier et al, 2015; Law et al, 2015)
Our study confirms the presences of V. parahaemolyticus in freshwater and marine fish samples in Selangor by having use highly accurate detection and identification method

Summary

Introduction

Vibrionaceae family within the class of Gammaproteobacteria comprises of Gram-negative halophilic bacteria, straight or curved rods, ubiquitous and indigenous in aquatic environments (Tison and Kelly, 1984; Tantillo et al, 2004; Sawabe et al, 2013). The species is widely distributed in marine and estuarine environments leading to gastrointestinal infections upon consumption of raw or undercooked seafood (Kubota et al, 2008; Letchumanan et al, 2014; Lee and Raghunath, 2018). 99% of clinical V. parahaemolyticus isolates are known to be pathogenic because they carry tdh genes and/or trh genes, whereas majority of the environmental isolates are non-pathogenic (Sudha et al, 2012; Tsai et al, 2013). Around 0–6% of the environmental isolates are identified as pathogenic carrying tdh gene and/or trh gene (Letchumanan et al, 2014, 2015a)

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