Document Type : Original Article
Department of Bacteriology, Animal Health Research Institute, Kafr El -Sheikh Branch, Agriculture Research Center, Giza, Egypt
Department of Microbiology, Faculty of Veterinary Medicine, Kafrelsheikh University, Kafr EL-Sheikh, Egypt
Department of Fish Diseases, Faculty of Veterinary Medicine, Kafrelsheikh University, Kafr EL-Sheikh, Egypt
Department of Biotechnology, Animal Health Research Institute, Dokki, Agriculture Research Center, Giza, Egypt
Vibrio species are significant pathogens affecting aquatic species. Around 12 species of Vibrio can cause a gastrointestinal illness (gastroenteritis) in humans resulting from eating contaminated food such as raw or undercooked shellfish. The indiscriminate use of antibiotics accelerates the development of resistance representing a severe challenge for controlling Vibrio outbreaks. In this study, the antibiotic resistance profile and the prevalence of pathogenic Vibrio species of apparently healthy and diseased fishes isolated from different types of fish in Kafr EL-Sheikh Governorate in Egypt during 2018 were determined. Samples obtained from fishes were inoculated onto a Vibrio-selective medium (TCBS) and phenotypically identified using the biochemical characteristics and representative cultures were checked by PCR to confirm the identified isolates. In the present study, V. alginolyticus (16.00%) was the predominant species followed by V. cholerae (7.33%) and V. parahaemolyticus (5.33%). The tested isolates were resistant to ampicillin (80.00%) and sensitive to ciprofloxacin and norfloxacin (100%). A total number of 15 Vibrio isolates (5 Vibrio parahaemolyticus, 5 V. alginolyticus, and 5 V. cholerae) were screened for five housekeeping genes and pathogenic virulence markers by PCR. Results showed that 100% of the V. parahaemolyticus isolates carried the tlh gene and 60.00% carried the tdh gene. In V. alginolyticus, 100% of the isolates carried the collagenase gene 0.00% carry the tdh gene; and 80.00% of V. cholerae isolates carried the ctx gene. The results showed that many isolates in this study had virulence characteristics that might correspond with the potential of infections and diseases.
- Abdelaziz M, Ibrahem MD, Ibrahim MA, et al. Monitoring of different vibrio species affecting marine fishes in Lake Qarun and Gulf of Suez: Phenotypic and molecular characterization. Egypt J Aquat Res 2017; 43(2): 141-146.
- Elliot EL, Kaysner CA, Jackson L, et al. Vibrio cholerae, V. parahaemolyticus, vulnificus, and other Vibrio spp. In: FDA Bacteriological Analytical Manual. 7th ed. Arlington, USA: Association of Official Analytical Chemists International 1992: 111-140.
- Rameshkumar P, Nazar AKA, Pradeep MA, et al. Isolation and characterization of pathogenic Vibrio alginolyticus from sea cage cultured cobia (Rachycentron canadum (Linnaeus 1766)) in India. Lett Appl Microbiol 2017; 65(5): 423-430.
- Fouz B, Alcaide E, Barrera R, et al. Susceptibility of Nile Tilapia (Oreochromis niloticus) to vibriosis due to Vibrio vulnificus biotype 2 (serovar E). Aquaculture 2002; 212(1-4): 21-30.
- Xu Y, Wang C, Zhang G, et al. ISCR2 is associated with the dissemination of multiple resistance genes among Vibrio spp. and Pseudoalteromonas spp. isolated from farmed fish. Arch Microbiol 2017; 199(6): 891-896.
- Aguirre-Guzmán G, Mejia Ruíz H, Ascencio F. A review of extracellular virulence product of Vibrio species important in diseases of cultivated shrimp. Aquac Res 2004; 35(15): 1395-1404.
- Nakhamchik A, Wilde C, Rowe-Magnus DA. Cyclic-di-GMP regulates extracellular polysaccharide production, biofilm formation, and rugose colony development by Vibrio vulnificus. Appl Environ Microbiol 2008; 74(13): 4199-4209.
- Rønneseth A, Castillo D, D'Alvise P, et al. Comparative assessment of Vibrio virulence in marine fish larvae. J Fish Dis 2017; 40(10): 1373-1385.
- Frans I, Michiels CW, Bossier P, et al. Vibrio anguillarum as a fish pathogen: virulence factors, diagnosis and prevention. J Fish Dis 2011; 34(9): 643-661.
- Austin B, Zhang XH. Vibrio harveyi. A significant pathogen of marine vertebrates and invertebrates. Lett Appl Microbiol 2006; 43(2): 119-124.
- Nor Najwa M, Daniel AMD, Amin M, et al. Detection of virulence genes in Vibrio alginolyticus isolated from green mussel, Perna viridis. Jurnal Teknologi (Sciences & Engineering) 2015; 77(25): 19-23.
- Feldhusen F. The role of seafood in bacterial food borne diseases. Microbes Infect 2000; 2(13):
- Nordstrom JL, Vickery MC, Blackstone GM, et al. Development of a multiplex real-time PCR assay with an internal amplification control for the detection of total and pathogenic Vibrio parahaemolyticus bacteria in oysters. Appl Environ Microbiol 2007; 73 (18): 5840-5847.
- DePaola A, Nordstrom JL, Bowers JC, et al. Seasonal abundance of total and pathogenic Vibrio parahaemolyticus in Alabama oysters. Appl Environ Microbiol 2003; 69(3): 1521-1526.
- Rodrigues de Menezes FG, da SilvaNeves S, Viana de Sousa O, et al. Detection of virulence genes in environmental strains of Vibrio cholerae from Estuaries in northeastern Brazil. Rev Inst Med Trop Sao Paulo 2014; 56(5): 427-432.
- Noga EJ. Fish disease: diagnosis and treatment 2nd Singapore City, Singapore: Wiley-Blackwell; 2010. http://dx.doi.org/10.1002/9781118786758.
- Baumann P, Schubert RHW. Family II. Vibrionaceae, Bergey's manual of systematic bacteriology. 1st Baltimore, USA: Williams & Wilkins Co. 1984; 516-550.
- Elliot EL, Kaysner CA, Jackson L. Vibrio cholerae, parahaemolyticus, V. vulnificus and other Vibrio spp. In: FDA Bacteriological Analytical Manual. 8th ed. Gaithersburg, USA: AOAC International 1995:
- Balouiri M, Sadiki M, and Ibnsouda SK. Methods for in vitro evaluating antimicrobial activity: A review. J Pharm Anal 2016; 6(2): 71-79.
- Di pinto A, Ciccarese G, Tantillo G, et al. A collagenase-targeted multiplex PCR assay for identification of Vibrio alginolyticus, Vibrio cholerae, and Vibrio para-haemolyticus. J Food Prot 2005; 68(1): 150-
- Gutierrez West CK, Klein SL, Lovell CR. High frequency of virulence factor genes tdh, trh, and tlh in Vibrio para-haemolyticus strains isolated from a Pristine Estuary. Appl Environ Microbiol 2013; 79(7): 2247-2252.
- Mousavi SL, Rasooli I, Nazarian Sh, et al. Simultaneous detection of Escherichia coli O157:H7, toxigenic Vibrio cholerae, and Salmonella typhimurium by multiplex PCR. Iran J Clin Infect Dis 2009; 4(2): 97-103.
- Mustapha S, Mustapha EM, Nozha C. Vibrio Alginolyticus: an emerging pathogen of foodborne diseases. Int J Sci Technol 2013; 2(4): 302-309.
- Smith KF, Thia J, Gemmill CEC, et al. Barcoding of the cytochrome oxidase I (COI) indicates a recent introduction of Ciona savignyi into New Zealand and provides a rapid method for Ciona species discrimination. Aquatic Invasions 2012; 7(3): 305-313.
- Abdel-Aziz M, Eissa AE, Hanna M, et al. Identifying some pathogenic Vibrio/Photobacterium species during mass mortalities of cultured Gilthead seabream (Sparus aurata) and European seabass (Dicentrarchus labrax) from some Egyptian coastal provinces. Int J Vet Sci Med 2013; 1(2): 87-95.
- Al-Taee AMR, Khamees NR, Al-Shammari NAH. Vibrio species isolated from farmed fish in Basra city in Iraq. J Aquac Res Dev 2017; 8: 2. 1000472. doi: 10.4172/ 2155-9546.1000472.
- Deng Y, Xu L, Chen H, et al. Prevalence, virulence genes, and antimicrobial resistance of Vibrio species isolated from diseased marine fish in South China. Sci Rep 2020; 10: 14329. doi: 10.1038/s41598-020-71288-0.
- Ahmed HA, El Bayomi RM, Hussein MA, et al. Molecular characterization, antibiotic resistance pattern and biofilm formation of Vibrio parahaemolyticus and cholerae isolated from crustaceans and humans. Int J Food Microbiol 2018; 274: 31-37.
- El-Bouhy Z, El-Nobi G, El-Murr A, et al. Study on vibriosis in Mugil Capito in El-Dakahlia and Damitta governorates, Egypt. Abbassa Int J Aqua 2016; 9(1): 19-35.
- Okoh AI, Igbinosa Antibiotic susceptibility profiles of some Vibrio strains isolated from wastewater final effluents in a rural community of the Eastern Cape Province of South Africa. BMC Microbiol 2010; 10: 143. doi: 10.1186/1471-2180-10-143.
- Scarano C, Spanu C, Ziino G, et al. Antibiotic resistance of Vibrio species isolated from Sparus aurata reared in Italian mariculture. New Microbiol 2014; 37(3): 329-337.
- Xie ZY, Hu CQ, Chen C, et al. Investigation of seven Vibrio virulence genes among Vibrio alginolyticus and Vibrio parahaemolyticus strains from the coastal mariculture systems in Guangdong, China. Lett Appl Microbiol 2005; 41(2): 202-207.
- Lee LH, Ab Mutalib NS, Law JW, et al. Discovery on antibiotic resistance patterns of Vibrio para-haemolyticus in Selangor reveals carbapenemase producing Vibrio parahaemolyticus in marine and freshwater fish. Front Microbiol 2018; 9, 2513. doi: 10.3389/fmicb.2018.02513.
- Alipour M, Issazadeh K, Soleimani J. Isolation and identification of Vibrio parahaemolyticus from sea-water and sediment samples in the southern coast of the Caspian Sea. Comp Clin Path 2012; 23(1): 129-133.
- Rojas MV, Matté MH, Dropa M, et al. Characterization of Vibrio Parahaemolyticus isolated from oysters and mussels in São Paulo, Brazil. Rev Inst Med Trop Sao Paulo 2011; 53(4): 201-205.
- Safa A, Jime JS, Shahel F. Cholera toxin phage: structural and functional diversity between Vibrio cholerae AIMS Microbiol 2020; 6(2): 144-151.
- Hounmanou YM, Mdegela RH, Dougnon TV, et al. 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 Res Notes 2016; 9(1): 466. doi: 10.1186/s13104-016-2283-0.
- Awasthi SP, Chowdhury N, Neogi SB, et al Development of a multiplex PCR assay for the detection of major virulence genes in Vibrio cholera including non-O1 and non-O139 serogroups. J Microbiol Methods 2019; 157; 54-58.
- Mustapha S, Mustapha EM, Brahim B, et al. Characterization of Vibrio alginolyticus trh positive from Mediterranean environment in Tamouda Bay (Morocco). World Environ 2012; 2(4): 76-80.
- Avsever ML. First report of trh positive Vibrio alginolyticus isolates from bivalve molluscs in Turkey. Rev Med Vet (Toulouse) 2016; 167(3/4): 65-70.
- Gargouti AS, Ab-Rashid MNK, Ghazali MF, et al. Detection of tdh and trh toxic genes in Vibrio Alginolyticus strain from mantis shrimp (Oratosquilla Oratoria). J Nutr Food Sci 2015; 5(5): 1000405.4172/ 2155-9600.1000405.