Kültür Levreklerinde (Dicentrarchus labrax) Vibrio parahaemolyticus Enfeksiyonu
Yıl 2019,
Cilt: 4 Sayı: 2, 104 - 110, 31.08.2019
Fikri Balta
,
Hasret Yılmaz
Öz
Bu çalışmanın amacı, Türkiye’nin Doğu Karadeniz Bölgesinde yüzer ağ
kafeslerde yetiştirilen levrek balıklar (Dicentrarchus
labrax, L. 1758)’nda görülen hastalık salgınlarından izole edilen suşların
tanımlanması gerçekleştirilmiştir. Hastalık etkenlerine karşı antibiyotiklerin
profilleri belirlenmiştir. Tipik hastalık semptomları gösteren levrek
balıkların böbrek ve dalaklarından %1,5 tuz içeren triptik soy agara (%1,5
NaCl-TSA=T-TSA) ekimleri yapılmış ve 20±1°C de 48 saat soğutmalı etüvde
inkübasyona bırakılmıştır. İzole edilen bakterileri tanımlamak için klasik
mikrobiyolojik testler, API 20E hızlı test kitleri ve PZR metodu kullanılarak
gerçekleştirilmiştir. Bakteri izolatları steril %1,5 tuzlu suda McFarland 0,5
standart bulanıklığında APİ 20E test kitlerine inokulasyonu yapılmıştır. PZR
testi V. parahaemolyticus tespit
etmek için 16S rRNA gen primerleri kullanılarak gerçekleştirilmiştir.
İzolatların hepsi kanlı agarda (%5 koyun kanlı ilaveli) β homoliz oluşturduğu
ve %7 NaCl içeren peptonlu suda iyi ürediği tespit edilmiştir. İzolatların
hepsi mikrobiyolojik testler ve API 20E test kiti sonuçlarına göre V. parahaemolyticus olduğu tespit edilmiştir.
Universal 16S rRNA primerleri kullanılarak
yapılan PZR test sonucuna göre suşların %98 V. parahaemolyticus olduğu doğrulanmıştır. Antibiyogramı yapılan izolatlar sulphamethoxazole %100, ampicilline %84,4, eritromisine
%71,9, oksitetrasiklin %62,5, trimetoprim-sulfametoksazol
%56,3 ve streptomisin %46,9 dirençli, fakat
suşların hepsi oksolinik asit, enrofloksasine ve florfenikol’e duyarlı olduğu
tespit edildi
Kaynakça
- Abdelaziz, M., Ibrahem, M.D., Ibrahim M.A., Abu-Elala N.M. & Abdel-moneam, D.A. (2017). Monitoring of different vibrio species affecting marine fishes in Lake Qarun and Gulf of Suez: Phenotypic and molecular characterization. Egyptian Journal of Aquatic Research, 43, 141-146.
- Austin, B. & Austin, D.A. (1987). Vibrios. 263-296. In: Austin B, Austin DA (Ed), Bacterial Fish Pathogens. Ellis Horwood Ltd., Chichester, UK.
- Austin, B. & Austin, D.A. (1999). Characteristics of the disease: gram-negative bacteria, Vibrionaceae representatives. 29-30, 108-115, 313-332. In: Austin B, Austin DA (Ed), Bacterial Fish Pathogens: Disease of Farmed and Wild Fish. Springer-Praxis Publishing, Chichester, UK.
- Austin, B. & Austin, D.A. (2007). Antimicrobial compounds. 379-385. In: Austin B, Austin DA (Ed), Bacterial Fish Pathogens: Diseases of Farmed and Wild Fish. Praxis Publishing Chichester, UK.
- Austin, B. & Austin, D.A. (2016). Vibrionaceae representatives. 357-411. In: Austin B, Austin DA (Ed), Bacterial Fish Pathogens: Disease of farmed and wild fish. Springer, New York.
- Balta, F. & Dengiz Balta, Z. (2016). Deniz Suyuna Nakledilen Gökkuşağı Alabalığı (Oncorhynchus mykiss) Yavrularında Görülen Vibrio Enfeksiyonu ve Tedavisi. Anadolu Çevre ve Hayvancılık Bilimleri Dergisi, 1(1), 14-20.
- Balta, F. & Dengiz Balta, Z. (2017). Doğu Karadeniz’de yetiştiriciliği yapılan gökkuşağı alabalıkları (Oncorhynchus mykiss)’ndan izole edilen Vibrio anguillarum suşlarının serotiplendirilmesi, genetik karakterizasyonu ve antimikrobiyal duyarlılığının belirlenmesi. Ankara Universitesi Veteriner Fakultesi Dergisi, 64, 321-328.
- Balta, F. (2016). Phenotypic, serotypic and genetic characterization and antimıcrobial susceptibility determination of Vibrio anguillarum, isolated from cultured sea bass (Dicentrarchus labrax L., 1758) in the southeast Black Sea, Turkey. Fresenius Environmental Bulletin, 25(10), 4393-4400.
- Balta, F., Sandalli, C., Kayis, S. Ozgumus, O.B. (2010). Molecular analysis of antimicrobial resistance in Yersinia ruckeri strains isolated from rainbow trout (Oncorhynchus mykiss) grown in commercialfish farms in Turkey. Bull Eur Ass Fish Pathol, 30(6), 211-219.
- Chakraborty, R.D. & Surendran, P.K., (2008). Occurrence and distribution of virulent strains of Vibrio parahaemolyticus in seafoods marketed from Cochin (India). World J Microbiol Biotechnol, 24, 1929-1935.
- CLSI. (2014). Performance standards for antimicrobial susceptibility testing. Twenty-Fourth Informational Supplement. Clinical Laboratory Standards Institute, Wayne, USA, M100-S24, 230s.
- Demircan, D. & Candan, A. (2006). Identification of Vibrio anguillarum by PCR (rpoN gene) associated with vibriosis in marine fish in Turkey. Turk J Vet Anim Sci, 30, 305-310.
- Drancourt, M., Bollet, C., Carlioz, A., Martelin, R., Gayral, J.P. & Raoult, D. (2000). 16S ribosomal DNA sequence analysis of a large collection of environmental and clinical unidentifiable bacterial isolates. J Clin Microbiol, 38, 362-3630.
- Frans, I., Michiels, C.W., Bossier, P., Willems, K.A., Lievens, B. & Rediers, H. (2011). Vibrio anguillarum as a fish pathogen: virulence factors, diagnosis and prevention. J Fish Dis, 34, 643-661.
- Grisez, L., Ceusters, R. & Oliever, F. (1991). The use of API 20E for the identification of Vibrio anguillarum and V. ordalii. J Fish Dis, 14, 359-365.
- Kent, M.L. (1982). Characteristics and identification of Pasteurella and Vibrio species pathogenic to fish using API 20 E (Analytab products) multitube test strips. Can J Fish Aquat Sci, 39, 1725-1729.
- Korun, J. & Gokoglu, M. (2007). Listonella anguillarum isolated from hatchery cultured red porgy Pagrus pagrus in Turkey. J Anim Vet Adv, 6, 823-827.
- Korun, J. (2006). Kültürü yapılan çipuralarda (Sparus aurata L.) görülen Listonella anguillarum enfeksiyonu üzerine bir çalışma. EU Su Ürünleri Dergisi, 23, 259-263.
- MacDonell, M.T. & Colwell, R.R. (1985). Phylogeny of the Vibrionaceae, and recommendation for two new genera, Listonella and Shewanella. Syst Appl Microbiol, 6, 171-182.
- MacDonell, M.T., Singleton, F.L. & Hood, M.A. (1982). Diluent composition for use of API 20E in characterizing marine and estuarine bacteria. Appl Environ Microbiol, 44, 423-427.
- Pal, D.& Das, N.(2010). Isolation, identification and molecular characterization of Vibrio parahaemolyticus from fish samples in Kolkata. European Review for Medical and Pharmacological Sciences, 14, 545-549.
- Popovic, N.T., Coz-Rakovac, R. & Strunjak-Petrovic, I. (2007). Commercial phenotypic tests (api 20E) in diagnosis of fish bacteria: A review. Veterinarni Medicina, 52, 49-53.
- Samuelsson, O.B., Nerland, A.H., Jorgensen, T., Schroder, M.B., Svasand, I.T. & Bergh, Q. (2006). Viral and bacterial diseases of Atlantic cod, Gadus morhua L. their prophylaxis and treatment: a review. Dis Aquat Org, 71, 239-254.
- Sarjito, R., Radjasa, O.K., Sabdono, A., Prayitno, S.B. & Hutabarat, S. (2009). Phylogenetic diversity of the causative agents of vibriosis associated with groupers fish from Karimunjawa Islands, Indonesia. Curr. Res. Bacteriol., 2, 14-21.
- Tanrıkul, T.T., Cagirgan, H.& Toksen, E. (2004). Levreklerden (Dicentrarchus labrax L., 1758) izole edilen vibrio türlerinin API 20E yöntemiyle identifikasyonu. EÜ su Ürünleri Dergisi, 21, 243-247.
- Tanrikul, T.T. (2007). Vibriosis as an epizootic disease of rainbow trout (Onchorhynchus mykiss) in Turkey. Pak J Biol Sci, 10, 1733-1737.
- Terzi Gulel, G. & Martinez-Urtaza, J. (2016). Molecular characterizations of Vibrio parahaemolyticus in seafood from the Black Sea, Turkey. Letters in Applied Microbiology, 62(6), 494-500. https://doi.org/10.1111/lam.12579.
- Urakawa, H., Kita-Tsukamoto, K. & Ohwada, K. (1997). 16S rDNA genotyping PCR/RFLP (restriction fragment length polymorphism) analysis among the family Vibronaceae. FEMS Microbiology Letters, 152, 125-132.
- Uzun, E. & Ogut, H. (2015). The isolation frequency of bacterial pathogens from sea bass (Dicentrarchus labrax) in the Southeastern Black Sea. Aquaculture, 437, 30-37.
- Weisburg, W.G., Barns, S.M., Pelletier, D.A. & Lane, D.J. (1991). 16S ribosomal DNA amplification for phylogenetic study. Journal of Bacteriolgy, 173, 697-703.
- Zhang, X.H. & Austin, B. (2005). Haemolysins in vibrio species. Journal of Applied Microbiology, 98(5), 1011-1019. doi:10.1111/j.1365-2672.2005.02583.x.
- Zorrilla, I., Arijo, S., Chabrillon, M., Diaz, P., Martinez-Manzanares, E., Balebona, M.C. & Morinigo, M.A. (2003). Vibrio species isolated from diseased farmed sole, Solea senegalensis (Kaup), and evaluation of the potential virulence role of their extracellular products. Journal of Fish Disease, 26(2), 103-108.
Infection of Vibrio parahaemolyticus in culture sea bass (Dicentrarchus labrax)
Yıl 2019,
Cilt: 4 Sayı: 2, 104 - 110, 31.08.2019
Fikri Balta
,
Hasret Yılmaz
Öz
The aim of this study is to identify
of the isolated bacteria strains from disease outbreaks in cultured sea bass (Dicentrarchus labrax L., 1758) in
floating net cages in the Eastern Black Sea Region of Turkey. The profiles of antibiotics against disease agents were determined. The
kidney and spleen of diseased sea bass that showed typical disease symptoms was
streaked on the surface of tryptic soy agar (TSA) added with 1.5 % sodium
chloride, and was incubated in the cooled incubator at 20±1°C for 48 hours. To
identify of isolated bacteria was carried out by using conventional biochemical
tests, the API 20E system tests and the PCR method. All isolates were
inoculated in API 20E system test kits by adjusting to a turbidity matching a
0.5 McFarland standard in sterile 1.5% saline water. PCR assay was subjected to using a universal 16s rRNA gene
primers to detect V. parahaemolyticus. All isolates showed β hemolysis on blood agar (with 5% sheep blood),
and good growth in peptone water containing 7% NaCl. According to the 20E
system and conventional biochemical test results were determined that all
isolates were V. parahaemolyticus. All strains were confirmed as 98% V.
parahaemolyticus by using PCR assay with 16S rRNA. Results of the testing susceptibility
to antibiotics showed that V. parahaemolyticus isolates were resistant to 100% sulphamethoxazole, 84.4% ampicillin,
71.9% erythromycin, 62.5% oxytetracycline 56.3% trimethoprim-sulfamethoxazole
and 46.9% streptomycin, but all strains were found susceptible to oxolinic
acid, enrofloxacin and florphenicol.
Kaynakça
- Abdelaziz, M., Ibrahem, M.D., Ibrahim M.A., Abu-Elala N.M. & Abdel-moneam, D.A. (2017). Monitoring of different vibrio species affecting marine fishes in Lake Qarun and Gulf of Suez: Phenotypic and molecular characterization. Egyptian Journal of Aquatic Research, 43, 141-146.
- Austin, B. & Austin, D.A. (1987). Vibrios. 263-296. In: Austin B, Austin DA (Ed), Bacterial Fish Pathogens. Ellis Horwood Ltd., Chichester, UK.
- Austin, B. & Austin, D.A. (1999). Characteristics of the disease: gram-negative bacteria, Vibrionaceae representatives. 29-30, 108-115, 313-332. In: Austin B, Austin DA (Ed), Bacterial Fish Pathogens: Disease of Farmed and Wild Fish. Springer-Praxis Publishing, Chichester, UK.
- Austin, B. & Austin, D.A. (2007). Antimicrobial compounds. 379-385. In: Austin B, Austin DA (Ed), Bacterial Fish Pathogens: Diseases of Farmed and Wild Fish. Praxis Publishing Chichester, UK.
- Austin, B. & Austin, D.A. (2016). Vibrionaceae representatives. 357-411. In: Austin B, Austin DA (Ed), Bacterial Fish Pathogens: Disease of farmed and wild fish. Springer, New York.
- Balta, F. & Dengiz Balta, Z. (2016). Deniz Suyuna Nakledilen Gökkuşağı Alabalığı (Oncorhynchus mykiss) Yavrularında Görülen Vibrio Enfeksiyonu ve Tedavisi. Anadolu Çevre ve Hayvancılık Bilimleri Dergisi, 1(1), 14-20.
- Balta, F. & Dengiz Balta, Z. (2017). Doğu Karadeniz’de yetiştiriciliği yapılan gökkuşağı alabalıkları (Oncorhynchus mykiss)’ndan izole edilen Vibrio anguillarum suşlarının serotiplendirilmesi, genetik karakterizasyonu ve antimikrobiyal duyarlılığının belirlenmesi. Ankara Universitesi Veteriner Fakultesi Dergisi, 64, 321-328.
- Balta, F. (2016). Phenotypic, serotypic and genetic characterization and antimıcrobial susceptibility determination of Vibrio anguillarum, isolated from cultured sea bass (Dicentrarchus labrax L., 1758) in the southeast Black Sea, Turkey. Fresenius Environmental Bulletin, 25(10), 4393-4400.
- Balta, F., Sandalli, C., Kayis, S. Ozgumus, O.B. (2010). Molecular analysis of antimicrobial resistance in Yersinia ruckeri strains isolated from rainbow trout (Oncorhynchus mykiss) grown in commercialfish farms in Turkey. Bull Eur Ass Fish Pathol, 30(6), 211-219.
- Chakraborty, R.D. & Surendran, P.K., (2008). Occurrence and distribution of virulent strains of Vibrio parahaemolyticus in seafoods marketed from Cochin (India). World J Microbiol Biotechnol, 24, 1929-1935.
- CLSI. (2014). Performance standards for antimicrobial susceptibility testing. Twenty-Fourth Informational Supplement. Clinical Laboratory Standards Institute, Wayne, USA, M100-S24, 230s.
- Demircan, D. & Candan, A. (2006). Identification of Vibrio anguillarum by PCR (rpoN gene) associated with vibriosis in marine fish in Turkey. Turk J Vet Anim Sci, 30, 305-310.
- Drancourt, M., Bollet, C., Carlioz, A., Martelin, R., Gayral, J.P. & Raoult, D. (2000). 16S ribosomal DNA sequence analysis of a large collection of environmental and clinical unidentifiable bacterial isolates. J Clin Microbiol, 38, 362-3630.
- Frans, I., Michiels, C.W., Bossier, P., Willems, K.A., Lievens, B. & Rediers, H. (2011). Vibrio anguillarum as a fish pathogen: virulence factors, diagnosis and prevention. J Fish Dis, 34, 643-661.
- Grisez, L., Ceusters, R. & Oliever, F. (1991). The use of API 20E for the identification of Vibrio anguillarum and V. ordalii. J Fish Dis, 14, 359-365.
- Kent, M.L. (1982). Characteristics and identification of Pasteurella and Vibrio species pathogenic to fish using API 20 E (Analytab products) multitube test strips. Can J Fish Aquat Sci, 39, 1725-1729.
- Korun, J. & Gokoglu, M. (2007). Listonella anguillarum isolated from hatchery cultured red porgy Pagrus pagrus in Turkey. J Anim Vet Adv, 6, 823-827.
- Korun, J. (2006). Kültürü yapılan çipuralarda (Sparus aurata L.) görülen Listonella anguillarum enfeksiyonu üzerine bir çalışma. EU Su Ürünleri Dergisi, 23, 259-263.
- MacDonell, M.T. & Colwell, R.R. (1985). Phylogeny of the Vibrionaceae, and recommendation for two new genera, Listonella and Shewanella. Syst Appl Microbiol, 6, 171-182.
- MacDonell, M.T., Singleton, F.L. & Hood, M.A. (1982). Diluent composition for use of API 20E in characterizing marine and estuarine bacteria. Appl Environ Microbiol, 44, 423-427.
- Pal, D.& Das, N.(2010). Isolation, identification and molecular characterization of Vibrio parahaemolyticus from fish samples in Kolkata. European Review for Medical and Pharmacological Sciences, 14, 545-549.
- Popovic, N.T., Coz-Rakovac, R. & Strunjak-Petrovic, I. (2007). Commercial phenotypic tests (api 20E) in diagnosis of fish bacteria: A review. Veterinarni Medicina, 52, 49-53.
- Samuelsson, O.B., Nerland, A.H., Jorgensen, T., Schroder, M.B., Svasand, I.T. & Bergh, Q. (2006). Viral and bacterial diseases of Atlantic cod, Gadus morhua L. their prophylaxis and treatment: a review. Dis Aquat Org, 71, 239-254.
- Sarjito, R., Radjasa, O.K., Sabdono, A., Prayitno, S.B. & Hutabarat, S. (2009). Phylogenetic diversity of the causative agents of vibriosis associated with groupers fish from Karimunjawa Islands, Indonesia. Curr. Res. Bacteriol., 2, 14-21.
- Tanrıkul, T.T., Cagirgan, H.& Toksen, E. (2004). Levreklerden (Dicentrarchus labrax L., 1758) izole edilen vibrio türlerinin API 20E yöntemiyle identifikasyonu. EÜ su Ürünleri Dergisi, 21, 243-247.
- Tanrikul, T.T. (2007). Vibriosis as an epizootic disease of rainbow trout (Onchorhynchus mykiss) in Turkey. Pak J Biol Sci, 10, 1733-1737.
- Terzi Gulel, G. & Martinez-Urtaza, J. (2016). Molecular characterizations of Vibrio parahaemolyticus in seafood from the Black Sea, Turkey. Letters in Applied Microbiology, 62(6), 494-500. https://doi.org/10.1111/lam.12579.
- Urakawa, H., Kita-Tsukamoto, K. & Ohwada, K. (1997). 16S rDNA genotyping PCR/RFLP (restriction fragment length polymorphism) analysis among the family Vibronaceae. FEMS Microbiology Letters, 152, 125-132.
- Uzun, E. & Ogut, H. (2015). The isolation frequency of bacterial pathogens from sea bass (Dicentrarchus labrax) in the Southeastern Black Sea. Aquaculture, 437, 30-37.
- Weisburg, W.G., Barns, S.M., Pelletier, D.A. & Lane, D.J. (1991). 16S ribosomal DNA amplification for phylogenetic study. Journal of Bacteriolgy, 173, 697-703.
- Zhang, X.H. & Austin, B. (2005). Haemolysins in vibrio species. Journal of Applied Microbiology, 98(5), 1011-1019. doi:10.1111/j.1365-2672.2005.02583.x.
- Zorrilla, I., Arijo, S., Chabrillon, M., Diaz, P., Martinez-Manzanares, E., Balebona, M.C. & Morinigo, M.A. (2003). Vibrio species isolated from diseased farmed sole, Solea senegalensis (Kaup), and evaluation of the potential virulence role of their extracellular products. Journal of Fish Disease, 26(2), 103-108.