Isolation and identification by real time pcr and determination of some antibiotic resistance of Vibrio parahaemolyticus from shrimps
Abstract
This study was carried out to determine the presence of Vibrio parahaemolyticus (V. parahaemolyticus) from shrimps (n=90) obtained from fishermen in Aydın province. According to traditional methods, 40 isolates gave suspicious positive results on Thiosulfate Citrate Bile Sucrose (TCBS) agar. All V. parahaemolyticus strain known to have thermolabile hemolysin encoded by tlh gene as species marker. In order to identify suspicious strains, tlh genes specific to V. parahaemolyticus were detected by using tlh primers. Eleven strains of the suspicious isolates gave a positive reaction with Real Time PCR. Antibiotic resistance of the isolates gave a positive reaction as V. parahaemolyticus by real-time PCR, penicillin G (10units), clindamycin (2µg), piperacillin (100µg), amoxicillin-clavulanic acid (30µg), ciprofloxacin (5µg) and gentamicin (10µg) was determined by disk diffusion. All isolates showed resistance to Penicillin G and Clindamycin and One isolate showed multiple resistance to all antibiotics tested. All isolates, except the isolate showing multiple antibiotic resistance, were susceptible to Piperacillin, Ciprofloxacin and Gentamicin. As a result, it is recommended to establish effective control measures to reduce the risk of V. parahaemolyticus infection in terms of public health, it is recommended to use effective methods routinely to ensure the safety of foods and to detect V. parahaemolyticus in foods and the environment.
Keywords
Antibiotic resistance, public health, real time pcr, shrimp, tlh, Vibrio parahaemolyticus
References
- Baker-Austin C, Oliver JD, Alam M, Ali A, Waldor MK, Quadri F, Martinez-Urtaza J. (2018). Vibrio spp. infections. Nat Rev Dis Primers, 4(8), 1-19. https://doi.org/10.1038/s41572-018-0005-8
- Baron, S, Lesne, J, Jouy, E, Larvor, E, Kempf, I, Boncy, J, Rebaudet, S, Piarroux, R. (2016). Antimicrobial Susceptibility of Autochthonous Aquatic Vibrio cholerae in Haiti. Front Microbiol, 7, 1671. https://doi.org/10.3389/fmicb.2016.01671
- Bilung LM, Linang V, Apun K, Lihan S, Kqueen CH, Vincent M. (2015). Application of Real-Time PCR for the Detection and Enumeration of Vibrio parahaemolyticus from Seafood in Sarawak (Malaysia). Borneo J Resour Sci Technol, 5(2), 70–78.
- Clinical and Laboratory Standard Institute (CLSI). Performance Standards for Antimicrobial Susceptibility Testing; CLSI Document M100, 33th Edition. Wayne, PA: Clinical and Laboratory Standards Institute; 2023.
- Clinical and Laboratory Standard Institute (CLSI). Performance Standards for Antimicrobial Susceptibility Testing CLSI Supplement M100S. 26th ed. Wayne, PA: Clinical and Laboratory Standards Institute; 2016.
- Cruz, CD, Hedderley, D, Fletcher, GC. (2015). Long-term Study of Vibrio parahaemolyticus Prevalence and Distrubution in New Zealand Shellfish. Appl Environ Microbiol. 81, 2320–2327.
- Davis BJK, Jacobs JM, Davis MF, Schwab KJ, De Paola A, Curriero FC. Environmental determinants of Vibrio parahaemolyticus in the Chesapeake Bay. Appl Environ Microbiol 83:21, 2017. e01147-17. https://doi.org/10.1128/AEM.01147-17
- Diler, A, Ataş, Ş. (2003). Antalya Bölgesinden Avlanan Penaeus semisulcatus de Haan 1884’un Mikrobiyolojik ve Kimyasal Kalitesi ile Et Verimi. Turk J Vet Anim Sci. 27, 497–503.
- Doğruer Y, Telli AE. (2020). Determination of Vibrio parahaemolyticus in Seafoods Using Direct Plate Counting, Quantitative Loop-mediated İsothermal Amplification and Propidium Monoazide-qLAMP. Ankara Univ Vet Fak Derg, 67, 349–355.
- Eschbach, E, Martin, A, Huhn, J, Seidel, C, Heuer, R, Schumacher, JH, Ulrich, S, Axe, JO, Konietzny, A, Strauch, E, Oberheitmann, B. (2017). Detection of Enteropathogenic Vibrio parahaemolyticus, Vibrio cholerae, and Vibrio vulnificus: Performance of Real-Time PCR Kits in an İnter laboratory Study. Eur Food Res Technol, 243, 1335–1342. https://doi.org/10.1007/s00217-017-2844-z
