Maher, N., Ezzat, G., Darwish, S., Ragab, E. (2025). Molecular Insights and Virulence Genes Profiling of Some Vibrio Species Isolated from Commonly Consumed Seafood and Their Implications on Seafood Quality. Journal of Applied Veterinary Sciences, 10(3), 78-89. doi: 10.21608/javs.2025.375816.1587
Nesma Maher; Gehad A Ezzat; Samah F Darwish; Eman Ragab. "Molecular Insights and Virulence Genes Profiling of Some Vibrio Species Isolated from Commonly Consumed Seafood and Their Implications on Seafood Quality". Journal of Applied Veterinary Sciences, 10, 3, 2025, 78-89. doi: 10.21608/javs.2025.375816.1587
Maher, N., Ezzat, G., Darwish, S., Ragab, E. (2025). 'Molecular Insights and Virulence Genes Profiling of Some Vibrio Species Isolated from Commonly Consumed Seafood and Their Implications on Seafood Quality', Journal of Applied Veterinary Sciences, 10(3), pp. 78-89. doi: 10.21608/javs.2025.375816.1587
Maher, N., Ezzat, G., Darwish, S., Ragab, E. Molecular Insights and Virulence Genes Profiling of Some Vibrio Species Isolated from Commonly Consumed Seafood and Their Implications on Seafood Quality. Journal of Applied Veterinary Sciences, 2025; 10(3): 78-89. doi: 10.21608/javs.2025.375816.1587
Molecular Insights and Virulence Genes Profiling of Some Vibrio Species Isolated from Commonly Consumed Seafood and Their Implications on Seafood Quality
1Microbiology Department, Faculty of Veterinary Medicine, Cairo University, Egypt
2Food Hygiene and Control Department, Faculty of Veterinary Medicine, Cairo University, Egypt.
3Biotechnology Research Unit, Animal Reproduction Research Institute, Agricultural Research Centre, Egypt.
4Microbiology Department , Faculty of Veterinary Medicine, Cairo University, Egypt
Receive Date: 16 April 2025,
Revise Date: 20 May 2025,
Accept Date: 28 May 2025
Abstract
Seafood is a known reservoir for Vibrio species, some of which carry virulence factors and can be acquired via handling and cross-contamination in the retail markets, which may pose public health risks. This study investigated the prevalence, virulence-associated genes, and spoilage effects of Vibrio spp. isolated from 400 samples from eight seafood species collected in Egypt. Microbiological and molecular identification were conducted using conventional methods and confirmatory PCR assays targeting species-specific and virulence-associated genes. Vibrio spp. were detected in 51.75% of the samples, from which 66.6% were identified as virulent species: V. parahaemolyticus (33.3%), V. cholerae (22.2%), and V. vulnificus (11.1%). Gene screening revealed the presence of toxR and vvh in all V. parahaemolyticus and V. vulnificus isolates, while ctx and hylA were found in 60% and 10% of V. cholerae isolates, respectively. Quality assessments of contaminated seafood showed elevated levels of total volatile base nitrogen (TVB-N) and moderate changes in pH, indicating spoilage. Thiobarbituric acid reactive substances (TBA-RS) values remained within acceptable limits, suggesting limited lipid oxidation. These findings bring to light the public health and quality concerns associated with Vibrio-contaminated seafood and emphasize the need for routine monitoring and comprehensive risk assessments.
ABD-ELGHANY, S.M., and SALLAM, K.I., 2013. Occurrence and molecular identification of Vibrio parahaemolyticus in retail shellfish in Mansoura, Egypt. Food Control 33(2):399–405. https://doi.org/10.1016/j.foodcont.2013.03.024
AHMED, H.A., EL BAYOMI, R.M., HUSSEIN, M.A., KHEDR, M.H., REMELA, E.M.A., and El-ASHRAM, A.M., 2018. Molecular characterization, antibiotic resistance pattern and biofilm formation of Vibrio parahaemolyticus and V. cholerae isolated from crustaceans and humans. International journal of food microbiology, 274, pp.31-37. https://doi.org/10.1016/j.ijfoodmicro.2018.03.013
ALI, S.E., MAHANA, O., MOHAN, C.V., DELAMARE‐DEBOUTTEVILLE, J., and ELGENDY, M.Y., 2022. Genetic characterization and antimicrobial profiling of bacterial isolates collected from Nile tilapia (Oreochromis niloticus) affected by summer mortality syndrome. Journal of Fish Diseases, 45(12), pp.1857-1871. https://doi.org/10.1111/jfd.13710
AMARAL, R.A., PINTO, C.A., LIMA, V., TAVARES, J., MARTINS, A.P., FIDALGO, L.G., SILVA, A.M., GIL, M.M., TEIXEIRA, P., BARBOSA, J., and BARBA, F.J., 2021. Chemical-Based methodologies to extend the shelf life of fresh fish—A review. Foods, 10(10), p.2300. https://doi.org/10.3390/foods10102300
AWASTHI, S.P., CHOWDHURY, N., NEOGI, S.B., HINENOYA, A., HATANAKA, N., CHOWDHURY, G., RAMAMURTHY, T., and YAMASAKI, S., 2019. Development of a multiplex PCR assay for the detection of major virulence genes in Vibrio cholerae including non-O1 and non-O139 serogroups. Journal of microbiological methods, 157, pp.54-58. https://doi.org/10.1016/j.mimet.2018.12.012
BAKER-AUSTIN, C., OLIVER, J.D., ALAM, M., ALI, A., WALDOR, M.K., QADRI, F., and MARTINEZ-URTAZA, J., 2018.Vibrio spp. infections. Nature Reviews Disease Primers, 4(1), pp.1-19. https://doi.org/10.1038/s41572-018-0005-8
BELL, A., and BOTT, M., 2021. Vibriosis: what you and your patients need to know. Delaware Journal of Public Health, 7(1), p.14. https://doi.org/10.32481/djph.2021.001.005
BONNIN-JUSSERAND, M., COPIN, S., LE BRIS, C., BRAUGE, T., GAY, M., BRISABOIS, A., GRARD, T., and MIDELET-BOURDIN, G., 2019.Vibrio species involved in seafood-borne outbreaks (Vibrio cholerae, V. parahaemolyticus and V. vulnificus): Review of microbiological versus recent molecular detection methods in seafood products. Critical reviews in food science and nutrition, 59(4), pp.597-610. https://doi.org/10.1080/10408398.2017.1384715
CHOI, G., and CHOI, S.H., 2022. Complex regulatory networks of virulence factors in Vibrio vulnificus. Trends in microbiology, 30(12), pp.1205-1216.
DENG, Y., XU, L., CHEN, H., LIU, S., GUO, Z., CHENG, C., MA, H., and FENG, J., 2020. Prevalence, virulence genes, and antimicrobial resistance of Vibrio species isolated from diseased marine fish in South China. Scientific reports, 10(1), p.14329. https://doi.org/10.1038/s41598-020-71288-0
DI PINTO, A., CICCARESE, G., TANTILLO, G., CATALANO, D., and FORTE, V.T., 2005. A collagenase-targeted multiplex PCR assay for identification of Vibrio alginolyticus, Vibrio cholerae, and Vibrio parahaemolyticus. Journal of food protection, 68(1), pp.150-153. https://doi.org/10.4315/0362-028X-68.1.150
EL-BOUHY, Z., EL-NOBI, G., EL-MURR, A., and ABD El-HAKIM, S., 2016. Study on vibriosis in Mugil capito in El-Dakahlia and Damitta governorates, Egypt. Abbassa Int. J. Aquat, 9(1), p.2016.
EZZAT, G.A., KASSEM, G.M., and MALAK, N.M., 2025. Safety, quality attributes, and health risk assessments of sand smelt fish. Sci Rep., 15, 18278. https://doi.org/10.1038/s41598-025-03323-x
FREITAG, A., ELLETT, A., BURKART, H., and JACOBS, J., 2022. Estimating the economic burden of Vibrio parahaemolyticus in washington state oyster aquaculture: implications for the future. Journal of Shellfish Research, 40(3), pp.555-564. https://doi.org/10.2983/035.040.0312
GOBARAH, D.E.A., HELMY, S.M., MAHFOUZ, N.B., FAHMY, H.A., and ABOU, M.A.E.H.M., 2022, September. Virulence genes and antibiotic resistance profile of Vibrio species isolated from fish in Egypt. In Veterinary Research Forum (Vol. 13, No. 3, p. 315).https://doi.org/10.30466/vrf.2021.520767.3117
HOUNMANOU, Y.M., MDEGELA, R.H., DOUGNON, T.V., MHONGOLE, O.J., MAYILA, E.S., MALAKALINGA, J., MAKINGI, G., and DALSGAARD, A., 2016. Toxigenic Vibrio cholerae O1 in vegetables and fish raised in wastewater irrigated fields and stabilization ponds during a non-cholera outbreak period in Morogoro, Tanzania: an environmental health study. BMC Research Notes, 9, pp.1-8. https://doi.org/10.1186/s13104-016-2283-0
HUANG, Q., ZHANG, Y., ZHANG, M., LI, X., WANG, Q., JI, X., RONGRONG, C., XI, L., and SHENJIE, J. , RENFEI, L., 2024. Assessment of Vibrionaceae prevalence in seafood from Qidong market and analysis of Vibrio parahaemolyticus strains. PLoS ONE 19(8): e0309304. https://doi.org/10.1371/journal.pone.0309304
HUANG, Z., ANOKYEWAA, M.A., WANG, J., JIAN, J., and LU, Y., 2022. Pathogenicity and antibiotic resistance analysis of Vibrio species found in coastal water at mainly beach of Shenzhen, China. Frontiers in Marine Science, 9, p.980593.
HUBBARD, T.P., CHAO, M.C., ABEL, S., BLONDEL, C.J., ABEL ZUR WIESCH, P., ZHOU, X., DAVIS, B.M., and WALDOR, M.K., 2016. Genetic analysis of Vibrio parahaemolyticus intestinal colonization. Proceedings of the National Academy of Sciences, 113(22), pp.6283-6288. https://doi.org/10.1073/pnas.1601718113
HUSSEIN, M.A., EISSA, A.E., EL-TARABILI, R.M., ATTIA, A.S., ZAKI, M., IBRAHIM, T.B., ABDEL HADY, H.A., ABU MHARA, A., RAGAB, R.H., and DESSOUKI, A.A., 2024. Impact of Climate Change on Some Seasonal Bacterial Eruptions among Cultured Marine Fishes from Egyptian Coastal Provinces. Journal of Applied Veterinary Sciences, 9(2), pp.18-30. https://dx.doi.org/10.21608/javs.2024.253653.1298
IBRAHIM, M.A. 2017. Utilization of crayfish in some fishery products (Doctoral dissertation, Ph. D. thesis, Fac. Agric. Ain Shams Univ).
ISO (International Organization for Standardization), 2007a. ISO/TS 21872–2: 2007. Microbiology of food and animal feeding stuffs – Horizontal method for the detection of potentially enteropathogenic Vibrio spp. – Part 2: Detection of species other than Vibrio parahaemolyticus and Vibrio cholerae. https://www.iso.org/standard/38279.html
ISO (International Organization for Standardization), 2007b. ISO/TS 21872–1: 2007. Microbiology of food and animal feeding stuffs — horizontal method for the detection of potentially enteropathogenic Vibrio spp. —part 1: detection of Vibrio parahaemolyticus and Vibrio cholerae. https://www.iso.org/standard/38278.html
ISMAIL, E.M., KADRY, M., ELSHAFIEE, E.A., RAGAB, E., MORSY, E.A., RIZK, O., and ZAKI, M.M., 2021. Ecoepidemiology and potential transmission of Vibrio cholerae among different environmental niches: an upcoming threat in Egypt. Pathogens, 10(2), p.190. https://doi.org/10.3390/pathogens10020190
LALY, S.J., ANUPAMA, T.K., ASHOK KUMAR, K., SANKAR, T.V., and NINAN, G., 2021. Changes in biogenic amines, biochemical and microbial attributes of three spotted crab (Portunus sanguinolentus) during iced and refrigerated storage. Journal of Food Science and Technology, 58, pp.2197-2205. https://doi.org/10.1007/s13197-020-04730-w
LEE, E.J., and SHIN, H.S., 2019. Development of a freshness indicator for monitoring the quality of beef during storage. Food science and biotechnology, 28, pp.1899-1906. https://doi.org/10.1007/s10068-019-00633-5
LEE, S.E., SHIN, S.H., KIM, S.Y., KIM, Y.R., SHIN, D.H., CHUNG, S.S., LEE, Z.H., LEE, J.Y., JEONG, K.C., CHOI, S.H., and RHEE, J.H., 2000. Vibrio vulnificus has the transmembrane transcription activator ToxRS stimulating the expression of the hemolysin gene vvhA. Journal of bacteriology, 182(12), pp.3405-3415.https://doi.org/10.1128/jb.182.12.3405-3415.2000
LOO, K.Y., LAW, J.W.F., TAN, L.T.H., PUSPARAJAH, P., LETCHUMANAN, V., and LEE, L.H., 2022. Diagnostic techniques for rapid detection of Vibrio species. Aquaculture, 561, p.738628.https://doi.org/10.1016/j.aquaculture.2022.738628
MALAK, N. M., ABDEL-NAEEM, H. H., ABDELSALAM, A. A., and EZZAT, G. A., 2025. A comparative study concerning the sensory, physicochemical, bacteriological, nutritional quality, heavy metal content, and health risk assessment of some low-cost fish species. Food Control, 169, 111023. https://doi.org/10.1016/j.foodcont.2024.111023.
MALLE, P., and TAO, S.H., 1987. Rapid quantitative determination of trimethylamine using steam distillation. Journal of food Protection, 50(9), pp.756-760. https://doi.org/10.4315/0362-028X-50.9.756
MONTERO, D.A., VIDAL, R.M., VELASCO, J., GEORGE, S., LUCERO, Y., GÓMEZ, L.A., CARREÑO, L.J., GARCÍA-BETANCOURT, R., and O’RYAN, M., 2023. Vibrio cholerae, classification, pathogenesis, immune response, and trends in vaccine development. Frontiers in medicine, 10, p.1155751. https://doi.org/10.3389/fmed.2023.1155751
MORSHDY, A.M., THARWAT, A.E., and MAHMOUD, S.M., 2018. Chemical profile of salted and canned fish products. In 5th International Food Safety Conference Damanhur University Saturday, 13th October (pp. 556-568). https://doi.org/10.21608/ejas.2019.104437
NANDI, B., NANDY, R.K., MUKHOPADHYAY, S., NAIR, G.B., SHIMADA, T., and GHOSE, A.C., 2000. Rapid method for species-specific identification of Vibrio cholerae using primers targeted to the gene of outer membrane protein OmpW. Journal of clinical microbiology, 38(11), pp.4145-4151. https://doi.org/10.1128/jcm.38.11.4145-4151.2000
PANICKER, G., CALL, D.R., KRUG, M.J., and BEJ, A.K., 2004. Detection of pathogenic Vibrio spp. in shellfish by using multiplex PCR and DNA microarrays. Applied and environmental microbiology, 70(12), pp.7436-7444.https://doi.org/10.1128/AEM.70.12.7436-7444.2004
PARLAPANI, F.F., BOZIARIS, I.S., and DROSINOS, E.H., 2024. Detection of fish spoilage. In Handbook of seafood and seafood products analysis (pp. 560-585). CRC Press.
ROY, P., ROY, A., JEON, E., DEWITT, C., PARK, J., and PARK, S., 2024. Comprehensive analysis of predominant pathogenic bacteria and viruses in seafood products. Comprehensive Reviews in Food Science and Food Safety.;23(4):e13410. https://doi.org/10.1111/1541-4337.13410
SADAT, A., EL‐SHERBINY, H., ZAKARIA, A., RAMADAN, H., and AWAD, A., 2021. Prevalence, antibiogram and virulence characterization of Vibrio isolates from fish and shellfish in Egypt: A possible zoonotic hazard to humans. Journal of applied microbiology, 131(1), pp.485-498. https://doi.org/10.1111/jam.14929
SHARMIN, T., AKTER, M., BANIK, A., SUMIT, A.F., AHMED, T., MAHMUD, S., TALUKDER, A.H., and TALUKDER, K.A., 2021. Molecular Approaches for Isolation of Shigella Species from Culture Negative Diarrheal Samples: A Hospital Based Study in Bangladesh. American Journal of Biological and Environmental Statistics, 7(2), pp.39-43. https://doi.org/10.11648/j.ajbes.20210702.11
TAN, C.W., MALCOLM, T.T., KUAN, C.H., THUNG, T.Y., CHANG, W.S., LOO, Y.Y., PREMARATHNE, J.M., RAMZI, O.B., NORSHAFAWATIE, M.F., YUSRALIMUNA, N., and RUKAYADI, Y., 2017. Prevalence and antimicrobial susceptibility of Vibrio parahaemolyticus isolated from short mackerels (Rastrelliger brachysoma) in Malaysia. Frontiers in microbiology, 8, p.1087. https://doi.org/10.3389/fmicb.2017.01087
TARLADGIS, B.G., WATTS, B.M., YOUNATHAN, M.T., and DUGAN, L., 1960. A distillation method for the quantitative determination of malonaldehyde in rancid foods. Journal of the American Oil Chemists Society, 37, pp.44-48. https://doi.org/10.1007/BF02630824
WEI, S., ZHAO, H., XIAN, Y., HUSSAIN, M.A., and WU, X., 2014. Multiplex PCR assays for the detection of Vibrio alginolyticus, Vibrio parahaemolyticus, Vibrio vulnificus, and Vibrio cholerae with an internal amplification control. Diagnostic microbiology and infectious disease, 79(2), pp.115-118. https://doi.org/10.1016/j.diagmicrobio.2014.03.012
ZEIDLER, C., SZOTT, V., ALTER, T., HUEHN-LINDENBEIN, S., and FLEISCHMANN, S., 2024. Prevalence of Vibrio spp. in seafood from German supermarkets and fish markets. Foods, 13(24), p.3987. https://doi.org/10.3390/foods13243987
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