Original Articles
Vol. 13 No. 1 (2024)
https://doi.org/10.4081/ijfs.2024.11635

Detection of pathogenic Vibrio spp. in foods: polymerase chain reaction-based screening strategy to rapidly detect pathogenic Vibrio parahaemolyticus, Vibrio cholerae, and Vibrio vulnificus in bivalve mollusks and preliminary results

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Received: 3 August 2023
Accepted: 9 January 2024
Published: 26 February 2024
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V. parahaemolyticus, V. vulnificus, V. cholerae, real-time PCR

Authors

Orlandina Di Maro
https://orcid.org/0000-0001-5869-260X
Department of Food Inspection, Experimental Zooprophylactic Institute of Southern Italy, Portici, Italy.
Yolande T.R. Proroga
Department of Food Inspection, Experimental Zooprophylactic Institute of Southern Italy, Portici, Italy.
Silvia Castellano
https://orcid.org/0009-0002-5542-2625
Department of Food Inspection, Experimental Zooprophylactic Institute of Southern Italy, Portici, Italy.
Anna Balestrieri
https://orcid.org/0000-0001-6818-5628
Department of Food Inspection, Experimental Zooprophylactic Institute of Southern Italy, Portici, Italy.
Federico Capuano
Department of Food Inspection, Experimental Zooprophylactic Institute of Southern Italy, Portici, Italy.
Enrico Arletti
Generon S.p.A., San Prospero, Italy.
Michelangelo Vietina
Generon S.p.A., San Prospero, Italy.
Melissa Bizzarri
Generon S.p.A., San Prospero, Italy.
Nicoletta Murru
Department of Veterinary Medicine and Animal Production, University of Naples Federico II, Italy.
Maria Francesca Peruzy
mariafrancesca.peruzy@unina.it
Department of Veterinary Medicine and Animal Production, University of Naples Federico II, Italy.
Daniela Cristiano
https://orcid.org/0000-0001-8875-8200
Department of Food Inspection, Experimental Zooprophylactic Institute of Southern Italy, Portici, Italy.

The majority of human diseases attributed to seafood are caused by Vibrio spp., and the most commonly reported species are Vibrio parahaemolyticus, Vibrio vulnificus, and Vibrio cholerae. The conventional methods for the detection of Vibrio species involve the use of selective media, which are inexpensive and simple but time-consuming. The present work aimed to develop a rapid method based on the use of multiplex real-time polymerase chain reaction (PCR) to detect V. parahaemolyticus, V. vulnificus, and V. cholerae in bivalve mollusks. 30 aliquots of bivalve mollusks (Mytilus galloprovincialis) were experimentally inoculated with two levels of V. parahaemolyticus, V. vulnificus, and V. cholerae. ISO 21872-1:2017 was used in parallel for qualitative analysis. The limit of detection of 50% was 7.67 CFU/g for V. cholerae, 0.024 CFU/g for V. vulnificus, and 1.36 CFU/g for V. parahaemolyticus. For V. vulnificus and V. cholerae, the real-time PCR protocol was demonstrated to amplify the pathogens in samples seeded with the lowest and highest levels. The molecular method evaluated showed a concordance rate of 100% with the reference microbiological method. V. parahaemolyticus was never detected in samples contaminated with the lowest level, and it was detected in 14 samples (93.33%) seeded with the highest concentration. In conclusion, the developed multiplex real-time PCR proved to be reliable for V. vulnificus and V. cholerae. Results for V. parahaemolyticus are promising, but further analysis is needed. The proposed method could represent a quick monitoring tool and, if used, would allow the implementation of food safety.

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Ahmed A, Engelbert MFM, Boer KR, Ahmed N, Hartskeerl RA, 2009. Development and validation of a real-time PCR for detection of pathogenic Leptospira species in clinical materials. PLoS One 4:e7093.
Baker-Austin C, Oliver JD, Alam M, Ali A, Waldor MK, Qadri F, Martinez-Urtaza J, 2018. Vibrio spp. infections. Nat Rev Dis Primers 4:8.
Bustin SA, Mueller R, Nolan T, 2020. Parameters for successful PCR primer design. Methods Mol Biol 2065:5-22.
Castello A, Alio V, Sciortino S, Oliveri G, Cardamone C, Butera G, Costa A, 2022. Occurrence and molecular characterization of potentially pathogenic Vibrio spp. in seafood collected in Sicily. Microorganisms 11:53.
Chow KH, Ng TK, Yuen KY, Yam WC, 2001. Detection of RTX toxin gene in Vibrio cholerae by PCR. J Clin Microbiol 39:2594-7.
European Food Safety Authority, European Center for Disease Prevention and Control, 2022. The European Union one health 2021 zoonoses report. EFSA J 20:e07666.
Fleischmann S, Herrig I, Wesp J, Stiedl J, Reifferscheid G, Strauch E, Alter T, Brennholt N, 2022. Prevalence and distribution of potentially human pathogenic Vibrio spp. on German North and Baltic sea coasts. Front Cell Infect Microbiol 12:846819.
Garrido A, Chapela MJ, Ferreira M, Atanassova M, Fajardo P, Lago J, Vieites JM, Cabado AG, 2012. Development of a multiplex real-time PCR method for pathogenic Vibrio parahaemolyticus detection (tdh+ and trh+). Food Control 24:128-35.
Garrido-Maestu A, Chapela MJ, Peñaranda E, Vieites JM, Cabado AG, 2014. In-house validation of novel multiplex real-time PCR gene combination for the simultaneous detection of the main human pathogenic vibrios (Vibrio cholerae, Vibrio parahaemolyticus, and Vibrio vulnificus). Food Control 37:371-9.
Han F, Ge B, 2010. Quantitative detection of Vibrio vulnificus in raw oysters by real-time loop-mediated isothermal amplification. Int J Food Microbiol 142:60-6.
ISO, 2017a. Horizontal method for the determination of Vibrio spp. Part 1: detection of potentially enteropathogenic Vibrio parahaemolyticus, Vibrio cholerae and Vibrio vulnificus. Norm 21872-1:2017. International Standardization Organization ed., Geneva, Switzerland.
ISO, 2017b. Microbiology of the food chain. Horizontal method for the detection, enumeration and serotyping of Salmonella. Part 1: detection of Salmonella spp. Norm 6579-1:2017. International Standardization Organization ed., Geneva, Switzerland.
ISO, 2018. Microbiology of the food chain. Horizontal method for the enumeration of beta-glucuronidase-positive Escherichia coli. Part 1: colony-count technique at 44 degrees C using membranes and 5-bromo-4-chloro-3-indolyl beta-D-glucuronide. Norm 16649-1:2018. International Standardization Organization ed., Geneva, Switzerland.
ISO, 2023. Accuracy (trueness and precision) of measurement methods and results – part 1: general principles and definitions. Norm 5725-1:2023. International Standardization Organization ed., Geneva, Switzerland.
Kim JY, Lee JL, 2014. Multipurpose assessment for the quantification of Vibrio spp. and total bacteria in fish and seawater using multiplex real-time polymerase chain reaction. J Sci Food Agric 94:2807-17.
La Tela I, Peruzy MF, D’Alessio N, Di Nocera F, Casalinuovo F, Carullo MR, Cardinale D, Cristiano D, Capuano F, 2021. Serotyping and evaluation of antimicrobial resistance of salmonella strains detected in wildlife and natural environments in southern Italy. Antibiotics (Basel) 10:353.
Ma JY, Zhu XK, Hu RG, Qi ZZ, Sun WC, Hao ZP, Cong W, Kang YH, 2023. A systematic review, meta-analysis and meta-regression of the global prevalence of foodborne Vibrio spp. infection in fishes: a persistent public health concern. Mar Pollut Bull 187:114521.
Marceddu M, Lamon S, Consolati SG, Ciulli S, Mazza R, Mureddu A, Meloni D, 2017. Determination of Salmonella spp., E. coli VTEC, Vibrio spp., and Norovirus GI-GII in bivalve molluscs collected from growing natural beds in Sardinia (Italy). Foods 6:88.
Messelhäusser U, Colditz J, Thärigen D, Kleih W, Höller C, Busch U, 2010. Detection and differentiation of Vibrio spp. in seafood and fish samples with cultural and molecular methods. Int J Food Microbiol 142:360-4.
Peruzy MF, Proroga YTR, Capuano F, Corrado F, Santonicola S, De Medici D, Delibato E, Murru N, 2020. Detection and quantification of Campylobacter in foods: new analytic approaches to detect and quantify Campylobacter spp. in food samples. Ital J Food Saf 9:8591.
Savini F, Giacometti F, Tomasello F, Pollesel M, Piva S, Serraino A, De Cesare A, 2021. Assessment of the impact on human health of the presence of norovirus in bivalve molluscs: what data do we miss?. Foods 2021:2444.
Wang Y, Salazar JK, 2016. Culture-independent rapid detection methods for bacterial pathogens and toxins in food matrices. Compr Rev Food Sci Food Saf 15;183-205.

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1.
Detection of pathogenic Vibrio spp. in foods: polymerase chain reaction-based screening strategy to rapidly detect pathogenic Vibrio parahaemolyticus, Vibrio cholerae, and Vibrio vulnificus in bivalve mollusks and preliminary results. Ital J Food Safety [Internet]. 2024 Feb. 26 [cited 2026 Mar. 10];13(1). Available from: https://www.pagepressjournals.org/ijfs/article/view/11635
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