Sinop ilindeki hamsi ve zargana balıklarından Vibrio spp. izolasyonu ve karakterizasyonu

Year 2016, Volume: 73 Issue: 2, 121 - 130, 01.06.2016

Cumhur Avşar İsmet Berber Ahmet Kenan Yıldırım

Abstract

Amaç: Bu çalışmada, Sinop İlindeki balıkçılardan temin edilen hamsi ve zargana balık örneklerinden izole edilen Vibrio türlerinin karekterizasyonu ile bu izolatların antibiyotik dirençlilikleri, plazmid DNA ve SDS-PAGE sodyum dodesil sülfat poliakrilamid jel elektroforezis toplam hücre protein profillerinin belirlenmesi amaçlanmıştır.Yöntem: Balık örnekleri taze olarak alınmış, buz üzerinde korunarak laboratuvara getirilmiş ve hemen analize alınmıştır. Balık örnekleri 25 g tartılarak zenginleştirme amacıyla %3 NaCl içeren steril alkali peptonlu su 225 mL besiyerine ilave edilmiş ve 37 OC’de 18-24 saat inkübasyona bırakılmıştır. İnkübasyon sonrası Tiyosülfat Sitrat Safra Sükroz TCBS agar besiyerinde gelişen sarı, yeşil veya mavi-yeşil koloniler Tripton Soya Agar TSA besiyerine ekilmiş ve izolatların daha sonraki tanımlaması için bir gece süreyle inkübe edilmiştir. İzolatlar morfolojik, fizyolojik ve biyokimyasal testlere göre tanımlanmıştır. Bunlara ek olarak izolatların yedi farklı antibiyotiğe karşı dirençliliği disk-difüzyon yöntemiyle belirlenmiştir. Ayrıca, izolatların SDSPAGE hücresel protein ve plazmid DNA profilleri de çıkartılmıştır. Bulgular: Toplam 44 adet balık örneği Vibrio türlerinin belirlenmesine yönelik çalışılmış ve morfolojik, biyokimyasal İnceleme sonucunda; 12 adet Vibrio spp. belirlenmiştir. özellikleri yönünden incelenmiştir. properties. At the end of the analysis; 12 Vibrio species were determined. The isolates were resistant against to ceftazidime 66.6%, gentamisine 25%, tetracycline 16.6%, ceftriaxone 91.6%, amikacine 16.6%, ofloxacine 25% and penicillin G 58.3%. In addition, The MARI Multiple Antibiotic Resistance Index values of tested strains were found at the rate of 0.41. The numerical analysis of SDS-PAGE whole-cell protein profiles of Vibrio spp. isolates revealed two major clusters at similarity levels of 80% and above. The 1-4 plasmide range 1590 - 27000 bp in all of the isolates were determined.Conclusion: As a result, it is possible to say that microbial infections can be prevented by adequate freezing and cooking, proper storage and processing after harvesting and avoidance of cross-contamination during fish handling. In addition, the coastal contamination due to bacteria leads to decline in the quality of seawater resources and so leads to risks to human health and economic losses. Furthermore, our study indicated that the presence of antibiotic resistant Vibrio spp. strains isolated from fishes might be potential risk for health

Keywords

Vibrio, balık, SDS-PAGE, plazmid, antibiyogram

Isolation and characterization of Vibrio spp. from anchovy and garfish in the Sinop province

Abstract

Objective: In this study it was aimed to characterize the Vibrio species isolated from anchovy and garfish samples obtained from fishermen and to determine the antibiotic resistances, plasmide DNA and SDS-PAGE sodium dodecyl sulphate polyacrylamide gel electrophoresis whole cell protein profiles of these isolates.Method: The fish samples were taken as fresh, kept in ice during transport to the laboratory and immediately taken for analysis. The fish samples weighed as 25 grams were added into 225 mL of sterile alkaline peptone water supplemented with 3% NaCl broth for enrichment purpose and incubated at 37 OC for 18 to 24 hours. After incubation, yellow, green or blue-green colonies growing on Thiosulphate Citrate Bile Salt Sucrose TCBS agar were subcultured on Triptone Soya Agar TSA and incubated overnight for further identification of the isolates. The isolates were identified according to morphological, physiological and biochemical tests. Moreover, the resistances against seven different antibiotics of isolates were determined by disk-diffusion method. In addition, the SDS-PAGE cell protein and plazmid DNA profiles of isolates were determined.Results: Totallly 44 fish samples were tested for determining Vibrio species and were analyzed according to their morphological, biochemical properties. At the end of the analysis; 12 Vibrio species were determined. The isolates were resistant against to ceftazidime 66.6%, gentamisine 25%, tetracycline 16.6%, ceftriaxone 91.6%, amikacine 16.6%, ofloxacine 25% and penicillin G 58.3%. In addition, The MARI Multiple Antibiotic Resistance Index values of tested strains were found at the rate of 0.41. The numerical analysis of SDS-PAGE whole-cell protein profiles of Vibrio spp. isolates revealed two major clusters at similarity levels of 80% and above. The 1-4 plasmide range 1590 - 27000 bp in all of the isolates were determined.Conclusion: As a result, it is possible to say that microbial infections can be prevented by adequate freezing and cooking, proper storage and processing after harvesting and avoidance of cross-contamination during fish handling. In addition, the coastal contamination due to bacteria leads to decline in the quality of seawater resources and so leads to risks to human health and economic losses. Furthermore, our study indicated that the presence of antibiotic resistant Vibrio spp. strains isolated from fishes might be potential risk for health

Keywords

Vibrio, fish, SDS-PAGE, plasmide, antibiogram

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