Investigation of diseases caused by Aeromonas media in rainbow trout (Oncorhynchus mykiss) in commercial fish farms using MALDI-TOF and specification of antibiotic sensitivity profiles of the agent

• Filiz Özcan Dicle University, Veterinary Faculty, Fisheries and Fisheries Diseases Department, Diyarbakır, Turkey

DOI: https://doi.org/10.52973/rcfcv-e32191

Keywords: Aeromonas media, MALDI-TOF, Oncorhynchus mykiss

Abstract

Rainbow Trout (RT) Oncorhynchus mykiss Walbaum,1792 the first fish species to be grown intensively in Turkey and the most commonly grown species in dams and lakes today. Aeromonas spp. are found in ecosystems such as freshwater and coastal waters and cause disease in fish. In RT causes death that is characterized by internal organ hemorrhages, such as the spleen, kidney and liver and body surface, ulcerations and congestion. The present study aimed to detect Aeromonas media bacteria in fish samples collected from the RT farms in the South Eastern Anatolia Region of Turkey through bacteriological culturing and MALDI-TOF. Also, it was purposed to determine the antibiotic susceptibility of the agent. Antibiotic susceptibility testing was performed using the Kirby–Baüer disk diffusion method. Separate visits were made to Trout farming facilities in Diyarbakır, Adıyaman, Şanlıurfa, and Batman in January and February 2021, and 30 fish with an average live weight of 200–250 grams (g) were collected from each facility. A total of 1,200 samples were taken from 40 Trout farms. It was detected the causative agent in the liver, kidney, and tissue samples of fish from 28 of the 40 farms. The sensitivity of Enrofloxacin (10 microgram -µg-), Florfenicol (30 µg), Neomycin (10 µg), Amoxicillin (10 µg), Oxytetracycline (30 µg), Erythromycin (10 µg), Gentamycin (10 µg) and Streptomycin (10 µg) were defined at chancing ratios. In conclusion, these bacteria were detected in local farms. These farms must implement measures to minimize stress factors affecting the fish and avoid overstocking. The best methods of protection against infection include avoiding overstocking, low oxygen levels, and unskilled labor.

References

ALLEN, D.A.; AUSTIN, B.; COLWELL, R.R. Aeromonas media, a new species isolated from river water. Intern. J. System. Bacteriol. 33(3): 599–604. 1983.

AUSTIN, B. Methods for the diagnosis of bacterial fish diseases. Marine Life Sci. Tech. 1: 41–49. 2019.

AUSTIN, B.; AUSTIN, D.A. Aerobic Gram-Positive Rods and Cocci. Bacterial fish pathogens: disease of farmed and wild fish. 6th. Ed. Springer, Dordrecht. 552 pp. 2016.

BAUER, A.U.; KIRBY, W.M.; SHERRIS, J.C.; TRACK, M. Antibiotic susceptibility testing by standardized single disc method. J. Clin. Pathol. 45(4): 493–496.1966.

BALTA, F. Fırtına Deresindeki Gökkuşağı Alabalık Çiftliklerinde izole edilen Aeromonas spp. İzolatlarının Antimikrobiyel Hassasiyetin Belirlenmesi. J. Anatolian Environm. Anim. Sci. 5(3): 397–407. 2020.

BALCI, Ş. Aeromonas hydrophila ve Aeromonas sobria Bakterilerinin tanimlanmasinda farkli fenotipik tanimlama kitlerinin kullanilabilirliğinin karşilaştırılması, Recep Tayyip Erdoğan Üniversitesi Fen Bilimleri Enstitüsü Su Ürünleri Anabilim Dali Yüksek Lisans. Tezi. 31 pp. 2020.

BECTON-DICKINSON AND COMPANY (B.D). Antimicrobial susceptibility test discs. Loveton Circle Sparks, USA. Pp 16. 2011.

CLINICAL AND LABORATORY STANDARDS INSTITUTE (CLSI). Methods for antimicrobial disk susceptibility testing of bacteria isolated from aquatic animals; proposed guideline. M42-P, CLSI, Wayne, PA. 43pp. 2004.

ONUK, E.E.; ÇAYCI, Y.T.; ÇOBAN, A.Y.; ÇİFTCİ, A; BALTA, F.; DİDİNEN, B.I.; ALTUN, S. Balık ve yetiştirme suyu kökenli Aeromonas izolatlarının antimikrobiyal duyarlılıklarının saptanması. Ankara Üniv. Vet. Fak. Derg. 64: 69–73. 2017.

DUMAN, M. Gökkuşaği alabaliklarinda görülen motil aeromonas (A. hydrophila, A. sobria, A. caviae), Yersinia ruckeri ve Lactococcus garvieae bakterilerinin antimikrobiyal duyarliliklari ve duyarlilikta rol oynayan genlerin tespiti, Uludağ Üniversitesi Sağlik Bilimleri Enstitüsü Su Ürünleri Hastaliklari Anabilim Dali. Doktora Tezi. 116 pp. 2017.

ERER, H. Balık Hastalıkları. 2th. Ed. S. Ü. Basımevi, Konya. 133 pp. 2002.

FOOD AGRICULTURE ORGANIZATION OF THE UNITED NATIONS (FAO). The State of World Fisheries and Aquaculture, Food, Agriculture. 2022. On line: https://bit.ly/3rbD1VU. 18/04/2022.

FECKANINOVA, A.; KOSCOVA, J.; MUDRONOVA, D.; POPELKA, P.; TOROPILOVA, J. The use of probiotic bacteria against Aeromonas nfections in salmonid aquaculture. Aquacult. 469: 1–8. 2017.

FREIWALD, A.; SAUER, S. Phylogenetic classification and identification 462 of bacteria by mass spectrometry. Nat. Protoc. 4: 732–742. 2009.

HUANG, W.M.; WANG, L.; JI, G.Z.; WEI, X.; WANG, H.J.; ZHAO, J.; GOU, X.L; LONG, Z.M. Identification and drug sensitivity of bacteria Aeromonas veronii and A. media isolated from Chinese sucker Myxocyprinus asiaticus. Fisheries Sci. 32: 210–214. 2013.

IGBINOSA, I.H.; IGUMBOR, E.U.; AGHDASI, F.; TOM, M.; OKOH, A.L. Emerging Aeromonas Species Infections and Their Significance in Public Health. Sci. World J. 2012: 1–13. 2012.

KENZAKA, T.; NAKAHARA, M.; HIGUCHI, S.; MAEDA, K; TANI, K. Draft genome sequences of Amoeba-resistant Aeromonas spp. isolated from aquatic environments. Genome Announc. 2(5): e01115–14. 2014.

KILINÇ, B. Su ürünlerinde Aeromonas ve Plesiomonas cinsi mikroorganizmalar ve etkileri. Ege J. Fisheries Aquatic Sci. 36(2): 191–199. 2019.

LÜ, A.J.; HU, X C.; LI, L.; SUN, J.F.; SONG, Y.J.; PEI, C.; ZHANG, C.; KONG, X.H. Isolation, identification and antimicrobial susceptibility of pathogenic Aeromonas media isolated from diseased Koi carp (Cyprinus carpio koi). Iranian J. Fisheries Sci. 15(2): 760–774. 2016.

ÖZTÜRK, N. Bolu ilindeki alabalık üretim tesislerinde kullanılan ilaç ve kimyasalların araştırılması, Kocaeli Üniversitesi Fen Bilimleri Enstitüsü Su Ürünleri Anabilim Dalı Yüksek Lisans. Tezi. 39 pp. 2022.

HILBORN, R.; QUINN, T.; SCHINDLER, D.; ROGERS, D. Biocomplexity and fisheries sustainability. Proceed. Nat. Acad. Sci. United States of America. 100(11): 6564–6568. 2003.

RUANGPAN, L.; TENDENCIA, A.E. Laboratory manual of standardized methods for antimicrobial sensitivity tests for bacteria isolated from aquatic animals and environment. Aquaculture Extension Manual No 37. Southeast Asian Fisheries Development Center, Tigbauan. 5021, Iloilo, Philippines. 54 pp. 2004.

STOSKOPF, M.K. Nutrition and Nutritional Diseases. Fish Medicine. In: W.B. Saunders. Philadelphia. 882pp. 1993.

SHEN, X.J.; HU, X.C.; LAN, Y.; LI, X.; ZHU, A.H; ZHANG, P.; LV, A.J. Drug susceptibility testing and isolation and identification of Pseudomonas fluorescens from koi carp (Cyprinus carpio koi). Fisheries Sci. 33: 443–446. 2014.

TOPIĆ-POPOVIĆ, N.; KAZAZIĆ, S.; BOJANIĆ, K.; STRUNJAK-PEROVIĆ, I.; ČOŽ-RAKOVAC, R. Sample and culture conditions effects on MALDI-TOF MS identification of bacteria: A review. Vet. Glasnik. 2021: 1–8. 2021. https://doi.org/jdrm.

Türkish Statistical Institute (TUİK). 2020. Most important fish species. On Line: https://bit.ly/3SxF7ef. 22/06/2022.

TÜBA, I.I. Gıda ve Sağlıklı Beslenme Sempozyumu Raporu “Su Ürünleri ve Sağlık” 2019. Dr. Sahin, K. (Ed.). Ankara. On Line: https://bit.ly/3LIwJpW. 18/02/2022.

YILDIRIM, F. Afyonkarahisar, Antalya ve Eskişehir’den temin edilen baliklardan aeromonas salmonicida etkeninin izolasyonu ve identifikasyonu ile antibiyotik duyarliliğinin saptanmasi. Afyon Kocatepe Üniversitesi Sağlık Bilimleri Enstitüsü Mikrobiyoloji Ana Bilim Dalı. Degree Thesis. 33 pp. 2022.