Detection and quantification of pathogenic Vibrio parahaemolyticus in shellfish by using multiplex PCR and loop-mediated isothermal amplification assay

Malcolm, Tan Turk Hsern; Cheah, Yoke Kqueen; Radzi, Che Wan Jasimah Wan Mohamed; Kasim, Farihah Abu; Kantilal, Haresh Kumar; John, Tang Yew Huat; Martínez-Urtaza, Jaime; Nakaguchi, Yoshitsugu; Nishibuchi, Mitsuaki; Radu, Son

doi:10.1016/j.foodcont.2014.08.010

Cited by 48 publications

(40 citation statements)

References 32 publications

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“…Numerous prevalence studies and outbreaks of V. parahaemolyticus infection have been extensively reported in oysters, clams, cockles, mussels, crabs, and shrimps (Newton et al, 2014; Rodgers et al, 2014; Xu et al, 2014; Malcolm et al, 2015). In finfish species, qualitative detection of V. parahaemolyticus in the anchovies ( Engraulis spp.…”

Section: Introductionmentioning

confidence: 99%

Prevalence and Antimicrobial Susceptibility of Vibrio parahaemolyticus Isolated from Short Mackerels (Rastrelliger brachysoma) in Malaysia

et al. 2017

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Numerous prevalence studies and outbreaks of Vibrio parahaemolyticus infection have been extensively reported in shellfish and crustaceans. Information on the quantitative detection of V. parahaemolyticus in finfish species is limited. In this study, short mackerels (Rastrelliger brachysoma) obtained from different retail marketplaces were monitored with the presence of total and pathogenic strains of V. parahaemolyticus. Out of 130 short mackerel samples, 116 (89.2%) were detected with the presence of total V. parahaemolyticus and microbial loads of total V. parahaemolyticus ranging from <3 to >105 MPN/g. Prevalence of total V. parahaemolyticus was found highest in wet markets (95.2%) followed by minimarkets (89.1%) and hypermarkets (83.3%). Pathogenic V. parahaemolyticus strains (tdh+ and/or trh+) were detected in 16.2% (21 of 130) of short mackerel samples. The density of tdh+ V. parahaemolyticus strains were examined ranging from 3.6 to >105 MPN/g and microbial loads of V. parahaemolyticus strains positive for both tdh and trh were found ranging from 300 to 740 MPN/g. On the other hand, antibiotic susceptibility profiles of V. parahaemolyticus strains isolated from short mackerels were determined through disc diffusion method in this study. Assessment of antimicrobial susceptibility profile of V. parahaemolyticus revealed majority of the isolates were highly susceptible to ampicillin sulbactam, meropenem, ceftazidime, and imipenem, but resistant to penicillin G and ampicillin. Two isolates (2.99%) exhibited the highest multiple antibiotic resistance (MAR) index value of 0.41 which shown resistance to 7 antibiotics. Results of the present study demonstrated that the occurrence of pathogenic V. parahaemolyticus strains in short mackerels and multidrug resistance of V. parahaemolyticus isolates could be a potential public health concerns to the consumer. Furthermore, prevalence data attained from the current study can be further used to develop a microbial risk assessment model to estimate health risks associated with the consumption of short mackerels contaminated with pathogenic V. parahaemolyticus.

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Section: Introductionmentioning

confidence: 99%

Prevalence and Antimicrobial Susceptibility of Vibrio parahaemolyticus Isolated from Short Mackerels (Rastrelliger brachysoma) in Malaysia

et al. 2017

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“…The polymerase chain reaction (PCR) assays is one of the molecular techniques that is widely used to detect the presences of pathogenic V. parahaemolyticus strain in food and environment (Panicker et al, 2004;Yamamoto et al, 2008;Paydar et al, 2013;Malcolm et al, 2015). PCR primers can be multiplexed in a single reaction to increase the detection limit or tailored as real-time PCR analysis to provide more rapid results (Grant et al, 2006;Zhang et al, 2014).…”

Section: Introductionmentioning

confidence: 99%

Antimicrobial resistance profile and resistance genes of Vibrio species isolated from giant freshwater prawn (Macrobrachium rosenbergii) raised in Iran

Shakerian

Barton

Akinbowale

et al. 2018

J Hellenic Vet Med Soc

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Because of raising of large-scale high density prawn aquaculture techniques, production of this prawn worldwide is facing a serious threat from fatal diseases caused by nodaviruses and bacteria, particularly from the Vibrio species. The development of antibiotic resistance by Vibrio represents a potential threat to human health by exchange of resistant genes to human pathogens through food chain. This study aimed to determine antibiotic resistance profile of Vibrio isolates from giant fresh water Prawns (Macrobrachium rosenbergii) raised in Iran and to detect some antibiotic resistance genes in the isolates. A total of fifty giant fresh water prawns were processed for isolation of Vibrio species during February 2015 to August 2015. Identification of Vibrio isolates was done following standard biochemical methods. Phenotypic resistance of the isolates as determined by agar dilution method while polymerase chain reaction (PCR) method was used to detect the presence of erm, tetS, strA and sul2 genes in the isolates. Out of 50 prawns, 31 (62%) isolates of Vibrio spp. were reported, of which 20 (40%) were identified as V. parahaemolyticus, 10 (20%) were V. vulnificus and 1 (2%) were V. cholera. Over 90% of the tested strains showed susceptibility to SXT, AZM and NIT. In addition, strA and tetS genes were detected in all isolated Vibrio species. StrA gene was identified in 6 V. parahaemolyticus strains and also ermB and sul2 genes were not present in the isolate of V. cholera. The occurrence of multidrug resistance strains in the environment could be an indication of excessive usage of antibiotics in agriculture and aquaculture fields. This study has shown that giant freshwater prawns raised in Iran habour multidrug resistant Vibrio species.

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Section: Introductionmentioning

confidence: 99%

“…These methods were used to determine the total number of V. parahaemolyticus, which were then used to estimate the numbers of the pathogenic subtype (Di et al, 2015;Malcolm et al, 2015). Molecular techniques such as gene-specific probe and PCR are increasingly used to detect pathogenic V. parahaemolyticus defined by the presence of certain virulence markers (Malcolm et al, 2015). A loop-mediated isothermal amplification method has been recently developed and has since been coupled with other molecular techniques (Di et al, 2015;Malcolm et al, 2015;Notomi et al, 2015).…”

Section: Introductionmentioning

confidence: 99%

“…Molecular techniques such as gene-specific probe and PCR are increasingly used to detect pathogenic V. parahaemolyticus defined by the presence of certain virulence markers (Malcolm et al, 2015). A loop-mediated isothermal amplification method has been recently developed and has since been coupled with other molecular techniques (Di et al, 2015;Malcolm et al, 2015;Notomi et al, 2015). These newer DNA-based methods, in conjunction with the conventional biochemical tests often require the bacterial cells to be cultured on typical growth media first.…”

Section: Introductionmentioning

confidence: 99%

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Virulence gene expression of Vibrio parahaemolyticus in the viable but nonculturable state

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Detection and quantification of pathogenic Vibrio parahaemolyticus in shellfish by using multiplex PCR and loop-mediated isothermal amplification assay

Cited by 48 publications

References 32 publications

Prevalence and Antimicrobial Susceptibility of Vibrio parahaemolyticus Isolated from Short Mackerels (Rastrelliger brachysoma) in Malaysia

Prevalence and Antimicrobial Susceptibility of Vibrio parahaemolyticus Isolated from Short Mackerels (Rastrelliger brachysoma) in Malaysia

Antimicrobial resistance profile and resistance genes of Vibrio species isolated from giant freshwater prawn (Macrobrachium rosenbergii) raised in Iran

Virulence gene expression of Vibrio parahaemolyticus in the viable but nonculturable state

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