Aeromonas hydrophila Suşlarının Antibiyotik Direnç Profilleri

Year 2021, Volume: 17 Issue: 2, 202 - 213, 01.06.2021

Nurdan Filik Ebru Önem Ayşegül Kubilay

https://doi.org/10.22392/actaquatr.792224

Cited By: 1

Abstract

Aeromonas hydrophila virülens özellikleri açısından önemli bir patojen olup balıklarda ölümcül bir hastalık olan Motile Aeromonas Septisemisine (MAS) neden olur. Bu çalışmada hasta balıklardan izole edilen 20 A. hydrophila suşunun ve A. hydrophila ATCC 7966 suşunun 28 farklı antibiyotiğe karşı duyarlılığı araştırılmıştır. Araştırmada A. hydrophila suşlarında antibiyotik duyarlılık profilleri Kirby-Bauer disk difüzyon yöntemiyle saptanmış ve heri bir suşa ait Çoklu Antibiyotik Dirençliliği (ÇAD) indeksi hesaplanmıştır. Elde edilen sonuçlara göre suşların 12 antibiyotiğe dirençli, 14 antibiyotiğe duyarlı, 2 antibiyotiğe orta dercede duyarlı olduğu belirlenmiştir. ÇAD indeksi sonuçlarına göre suşların toplam 14 antibiyotiğe karşı çoklu antibiyotik direnci gösterdiği tespit edilmiştir. Suşların ÇAD indeks değerleri 0,21-0,75 aralığında tespit edilmiştir. Son yıllarda yapılan çalışmaların ortaya koyduğu bir bulgu A. hydrophila’nın oksitetrasikline karşı geliştirdiği dirençtir. Bu açıdan araştırma bulgularında en dikkat çeken oksitetrasiklin direnci olmuştur. A. hydrophila suşlarının araştırmada kullanılan antibiyotiklerin büyük bir kısmına ve yapılan bazı çalışmalarla tedavide en etkili antibiyotik olduğu bildirilen oksitetrasikline karşı direnç geliştirdiğinin tespit edilmesi suşların direnç profilinin yüksek olduğunu göstermektedir.
Sonuç olarak, balık patojenlerinin oluşturduğu hastalıkların tedavisinde yanlış antibiyotik kullanımı sonucu patojenin direnç kazandığı, bu durumunda balık hastalıkları ile mücadelede ciddi bir sorun oluşturduğu görülmektedir. 

Keywords

Aeromonas hydrophila, Balık, Antibiyogram, Çoklu Antibiyotik Dirençliliği, CLSI

Supporting Institution

Türkiye Bilimsel ve Teknolojik Araştırma Kurumu (TÜBİTAK)

Project Number

119O671

Thanks

Bu çalışma doktora tezinden özetlenmiştir. Türkiye Bilimsel ve Teknolojik Araştırma Kurumu (TÜBİTAK) 1002 Hızlı Destek Programı kapsamında 119O671 Proje Nosuyla desteklenmiştir. Araştırma da kullanılan A. hydrophila suşları Bursa Uludağ Üniversitesi (BUÜ)’nin, Isparta Uygulamalı Bilimler Üniversitesi (ISUBÜ)’nin, Ondokuz Mayıs Üniversitesi (OMÜ)’nin ve Recep Tayyip Erdoğan Üniversitesi (RTEÜ)’nin değerli Öğretim Elemanları tarafından temin edilmiş olup kendilerinin şahıslarına ve bağlı bulundukları Üniversitelerine teşekkürlerimizi sunarız.

Antibiotic Resistance Profiles of Aeromonas hydrophila Strains

Abstract

Aeromonas hydrophila virulent is a pathogen with important virulent properties and causes Motile Aeromonas Septicemia (MAS), a deadly disease in fish. In this study sensitivity of 20 A. hydrophila strains and A. hydrophila ATCC 7966 strains isolated from sick fish to 28 different antibiotics were investigated. In the study, antibiotic susceptibility profiles of A. hydrophila strains were determined by Kirby-Bauer disk diffusion method and the Multiple Antibiotic Resistance (MAR) index for each strain was calculated. According to the results, it was determined that the strains were resistant to 12 antibiotics, sensitive to 14 antibiotics, and 2 antibiotics moderately. According to the results of the MAR index, the strains were found to have multiple antibiotic resistance against the total of 14 antibiotics. MAR index values of strains were determined in the range of 0.21-0.75. One of the findings of recent studies is the resistance developed by A. hydrophila against oxytetracycline. In this respect, oxytetracycline resistance was the most striking in the research findings. The fact that A. hydrophila strains developed resistance to most of the antibiotics used in the research and oxytetracycline, which was reported to be the most effective antibiotic in treatment, shows that the resistance profile of strains is high.
As a result, it is seen that pathogen has gained resistance as a result of the use of broad-spectrum antibiotics in the treatment of diseases caused by fish pathogens, and this situation creates a serious problem in the fight against fish diseases.

Keywords

Aeromonas hydrophila, Balık, Antibiyogram, Çoklu Antibiyotik Dirençliliği, CLSI

Project Number

119O671

References

• Agger, W.A., McCormick, J.D., & Gurwith, M.J. (2018). Aeromonas hydrophila ilişkili ishalin klinik ve mikrobiyolojik özellikleri. Journal Clinical Microbiology.
• Aksoy, A. (2015). Antibiyotik Direnci Özel Sayısı. Türkiye Klinikleri Veteriner Bilimleri-Farmakoloji ve Toksikoloji-Özel Konular, 15(1).
• Akşit, A., & Kum, V. (2008). Gökkuşağı Alabalıkları (Oncorhynchus mykiss, Walbaum 1792)’nda sık görülen patojen mikroorganizmaların tespiti ve antibiyotik duyarlılık düzeylerinin belirlenmesi. Yüzüncü Yıl Üniversitesi Veteriner Fakültesi Dergisi, 19(1), 1-7.
• Alcaide, E., Blasco, M.D., & Esteve, C. (2005). Occurrence of drug-resistant bacteria in two European eel farms. Applied and Environmental Microbiology, 71(6), 3348–3350. https://doi.org/10.1128/AEM.71.6.3348-3350.2005
• Altwegg, M., & Jöhl, M. (1989). Isolation frequency of Aeromonas species in relation to patient age, Notes, 6, 55.
• Andersson, D. I., Balaban, N. Q., Baquero, F., Courvalin, P., Glaser, P., Gophna, U., ... & Tønjum, T. (2020). Antibiotic resistance: turning evolutionary principles into clinical reality. FEMS Microbiology Reviews, 44(2), 171-188.
• Angulo, F.J., & Griffin, P.M. (2000). Changes in antimicrobial resistance in Salmonella enterica serovar Typhimurium. Emerging Infectious Diseases Journal, 6, 436–438.
• Angulo, F.J., Nargund, V.N. & Chiller, T.C. (2004). Evidence of an association between use of anti-microbial agents in food animals and anti-microbial resistance among bacteria isolated from humans and the human health consequences of such resistance. Journal of Veterinary Medicine, 51(8-9), 374–379. https://doi.org/10.1111/j.1439-0450.2004.00789.x
• Austin, B., & Austin, D.A. (2016). Bacterial fish pathogens. 6. th edition. Springer International Publishing, Switzerland, pp: 21-82, 161-321, 323-396, 643-721.
• Ausubel, F., Brent, R., Kingston, R., Moor, D., Seidman, J., Smith, J., & Stauhle, K. (1988). Current Protocols in Molecular Biology. New York: Wiley Intersciences.
• Baran, I., Timur, M., Aydın, N., İstanbulluoğlu, E. and Aydintuğ, M.K. 1980. Çifteler-Sakaryabaşı balık üretim ve araştırma istasyonunda, alabalıklarda (Salmo gairdneri) görülen bakteriyel hemorajik septisemi hastalığı üzerine incelemeler. Ankara Üniversitesi Veteriner Fakültesi Dergisi, 27(1), 467-473.
• Boran, H., Terzi, E., Altinok, I., Capkin, E. and Bascinar, N. 2013. Bacterial diseases of cultured mediterranean Horse mackerel (Trachurus mediterraneus) in sea cages. Aquaculture, 396, 8-13.
• Cabello, F.C. (2006). Heavy use of prophylactic antibiotics in aquaculture: a growing problem for human and animal health and for the environment. Environmental microbiology, 8(7), (pp. 1137-1144). https://doi.org/10.1111/j.1462-2920.2006.01054.x
• Cappuccino, J.G., & Sherman, N. (1992). Biochemical activities of microorganisms. In: Microbiology, A Laboratory Manual. The Benjamin/Cummings Publishing Co. California, USA. 76 s.
• Castro, G. (2002). Characterisation of Aeromonas spp. isolated from frozen fish intended for human consumption in Mexico, International Journal of Food Microbiology, 2612, 1-9.
• Clark, N.M., & Chenoweth, C.E. (2003). Aeromonas infection of the hepatobiliary system: Report of 15 cases and review of the literature. Clinical Infectious Diseases, 37, 506-13.
• CLSI (Clinical and Laboratory Standards Institute), (2017). Performance standards for antimicrobial susceptibility testing. 27th ed., CLSI Supplement M100, USA.
• Costerton, J.W., Stewart, P.S., Greenberg, E.P. (1999). Bacterial Biofilms: A common Cause of Persistent Infections, Science, 284, 1318-1322.
• Davies, A., Capell, C., Jehanno, D., Nychas, G.J.E., & Kirby, R.M. (2001). Incidence of Foodborne Pathogens on Europen Fish, Food Control, 12, 67-71.
• Del Castillo, C. S., Hikima, J. I., Jang, H. B., Nho, S. W., Jung, T. S., Wongtavatchai, J., ... & Aoki, T. (2013). Comparative sequence analysis of a multidrug-resistant plasmid from Aeromonas hydrophila. Antimicrobial Agents, 57(1), 120-129.
• Duman, M. (2017). Gökkuşağı Alabalıklarında Görülen Motil Aeromonas (Aeromonas hydrophila, A. sobria, A. caviae), Yersinia ruckeri ve Lactococcus garvieae Bakterilerinin Antimikrobiyal Duyarlılıkları ve Duyarlılıkta Rol Oynayan Genlerin Tespiti. Doktora Tezi, T.C. Uludağ Üniversitesi Sağlık Bilimleri Enstitüsü.
• Durmaz, Y. & Türk, N. (2009). Alabalık İşletmelerinden Motil Aeromonasların İzolasyonu ve Antibiyotiklere Duyarlılıklarının Saptanması, Kafkas Üniversitesi Veteriner Fakültesi Dergisi, Research Article, 15(3), 357-361.
• Ehinmidu, J.O. (2003). Antibiotics Susceptibility Patterns of Urine Bacterial Isolates in Zaria, Nigeria. Tropical Journal of Pharmaceutical Research, 2(2), 223- 228. Erer, H. (2002). Balık Hastalıkları. 2. baskı, Selçuk Üniv. Basimevi, Konya.
• Gatesoupe, F.J. (1999). The use of probiotics in aquaculture. Aquaculture, 180, 147- 165.
• Gudding, R., Lillehaug, A., & Evensen, Ø., (1999). Recent developments in fish vaccinology. Veterinary Immunology and Immunopathology, 72, 203-212.
• Güvener, R.P. (2001). A study on the diagnosis of the aeromonad enfections in some aquarium fishes. Msc. thesis. Istanbul University.
• Heppell, J., & Davis, H.L. (2000). Application of DNA Vaccine Technology to Aquaculture. Advanced Drug Delivery Reviews, 43, 29-43.
• Hossain, S., Dahanayake, P.S., De Silva, B.C.J., Wickramanayake, M.V.K.S., Wimalasena, S.H.M.P., & Heo, G.J. (2019). Multidrug resistant Aeromonas spp. isolated from zebrafish (Danio rerio): antibiogram, antimicrobial resistance genes and class 1 integron gene cassettes. Letters in applied microbiology, 68(5), 370-377.
• Kaskhedikar, M., & Chhabra, D. (2010). Multiple drug resistance in Aeromonas hydrophila isolates of fish. Food Microbiology, 28, 157-168.
• Ko, W.C., Lee, H.C., Chuang, Y.C., Liu, C.C., & Wu, J.J. (2000). Clinical features and therapeutic implications of 104 episodes of monomicrobial Aeromonas bacteremia. Journal of Infectious, 40, 267-73.
• Korun, J., & Toprak, H.B. (2010). Kültürü yapılan gökkuşağı alabalıkları (Oncorhynchus mykiss)’nin bağırsağından izole edilen hareketli Aeromonas suşlarının antibiyotik hassasiyetleri üzerine NACl’un etkisi. Kafkas Üniversitesi Veteriner Fakültesi Dergisi, 16(2), 193-198.
• Krumperman, P.H. (1983). Multiple Antibiotic Resistance Indexing of Escherichia coli to Identify HighRisk Sources of Fecal Contamination of Food. American Society for Microbiology, Applied Environmental Microbiology, 46(1), 165-170.
• Kusdarwati, R., Kurniawan, H., & Prayogi, Y.T. (2017). Isolation and identification of Aeromonas hydrophila and Saprolegnia sp. on catfish (Clarias gariepinus) in floating cages in Bozem Moro Krembangan Surabaya. In IOP Conference Series: Earth and Environmental Science, 55(1) p. (012038). IOP Publishing.
• L’Abee-Lund, T.M., & Sorum, H. (2001). Class 1 integrons mediate antibiotic resistance in the fish pathogen Aeromonas salmonicida worldwide. Microbial Drug Resistance, 7(3), 263–272. https://doi.org/10.1089/10766290152652819.
• Laith, A.R., & Najiah, M. (2013). Aeromonas hydrophila: antimicrobial susceptibility and histopathology of isolates from diseased catfish, Clarias gariepinus (Burchell). Journal of Aquaculture Research and Development, 5(2), 1-7. https://doi.org/10.4172/2155-9546.1000215.
• Lasee, B. A., (1995). Introduction To Fish Healt Management, U.S. Fish and Wildlife Service La Crosse Fish Healt Center 555, Lester Avenue Onalaska, Wisconsin, 54650.
• Lehone, L., & Rawlin, G.P. (2000). Fish Diseases and Human Health in Aquaculture, Medical Journal of Australia, 173(5), 256-259.
• Lukistyowati, I. (2012). Pelacakan Gen Aerolysin dari Aeromonas hydrophila pada Ikan Mas yang Diberi Pakan Ekstrak Bawang Putih (Detection of Aerolysin Gen orom Aeromonas hydrophila In Common Carp Fed With Garlic Extract). Jurnal Veteriner Maret, 13(1), 43-50.
• Mancini, M.A., Galetto, M., & Gonzalez Quintana, H. (1997). Identification, clinical signs and histopathological lesions of Aeromonas hydrophila in fishes (Odontesthes bonariensis). Revista de Medicina Veterinaria Buenos Aires, 78(1), 65-68.
• Mathewson, J.J., & Dupont, H.L. (1992). Aeromonas species: role as human pathogens, In: Remington, J.S., Swartz, M.N. (eds.), Current Clinical Topics in Infectious Diseases, Vol12e. Cambridge: Blackwell Scientific. pp 26–36.
• Matyar, F. (2016). Hastane Kanalizasyonlarından İzole Edilen Gram-negatif Bakterilerin Tiplendirilmesi ve Çoklu Antibiyotik Dirençliliklerinin Saptanması. Türk Tarım – Gıda Bilim ve Teknoloji Dergisi, 4(10),845-849.
• Mueller, H.J. & Hinton, J. (1941). A protein-free medium for primary isolation of the Gonococcus and Meningococcus. Proceedings of the Society for Experimental Biology and Medicine, 48, 330-333.
• Naylor, R., & Burke, M. (2005) Aquaculture and ocean resources: raising tigers of the sea. Annual Review of Environment and Resources, 30,185–218.
• Onuk, E.E., Fındık, A., Turk, N., Altun, S., Korun, J., Ozer, S., Avsever, M.L. ve A. Ciftci, “Molecular identification and determination of some virulence genes of Aeromonas spp. in fish and water from Turkish coastal regions.” Revue de Medecine Veterinaire 164 (4): 200-206. (2013)
• Onuk, E.E., Tanrıverdi Çaycı, Y., Çoban, A.Y., Çiftci, A., Balta, F., Didinen, B.I., & Altun, S., (2017). Balık ve yetiştirme suyu kökenli Aeromonas izolatlarının antimikrobiyal duyarlılıklarının saptanması. Kısa Bilimsel Çalışma/Short Communication, Ankara Üniversitesi Veteriner Fakültesi Dergisi, 64, 69-73.
• Öztürk, R.Ç., & Altınok, İ. (2014). Bacterial and viral fish diseases in Turkey. Turkish Journal of Fisheries and Aquatic Sciences, 14(1), 275-297.
• Özturk, D., Adanır, R., & Turutoğlu, H. (2007). Bir sazan (Cyprinus carpio) işletmesinde Aeromonas hydrophila izolasyonu ve antibiyotik duyarlılığı. Ulusal Su Ürünleri Sempozyumu, at Mugla.
• Paul, S., Bezbaruah, R.L., Roy, M.K., & Ghosh, A.C. (1997). Multiple antibiotic resistance (MAR) index and its reversion in Pseudomonas aeruginosa. Letters in Applied Microbiology, 24, 169-171.
• Petersen, A, Andersen, J.S., Kaewmak, T., Somsiri, T., & Dalsgaard, A. (2002). Impact of integrated fish farming on antimicrobial resistance in a pond environment. Appliend and Environmental Microbiology, 68, 6036–6042. P oobalane, S., Thompson, K.D., Ardo, L., Verjan, N., Han, H.J., & Jeney, G. et. al. (2010). Production and efficacy of an Aeromonas hydrophila recombinant S-layer protein vaccine for fish. Vaccine, 28, 3540–7.
• Radu, S., Ahmad, N., Ling, F.H., & Reezal, A. (2003). Prevalence and resistance to antibiotics for Aeromonas species from retail fish in Malaysia. International Journal of Food Microbiology, 81(3), 261-266. https://doi.org/10.1016/s0168-1605(02)00228-3.
• Rasko, D., & Sperandio, V. (2010). Anti-Virulence Strategies to Combat Bacteria-Mediated Disease, Nature Reviews Drug Discovery, 9, 117-128. Rehulka, J. (2002). Aeromonas causes severe skin lesions in Rainbow trout, Clinical pathology, haemotology and biochemistry, Research Institute of Fish Culture and Hydrobiology, 71, 351-360.
• Roberts, R.J., & Shepherd, C.J. (2001). Alabalık ve Salmon Hastalıkları (Handbook of Trout and Salmon Diseases), (Çevirmen: Vatansever, H.). Akademi-UĞURER Tarımsal Kitap Tanıtım ve Pazarlama Hizmetleri, Kayseri, s.254.
• Sağlam, Y.S., Işık, N., Arslan, A., & Erer, H. (2006). Erzurum Bölgesindeki Gökkuşağı Alabalıklarında (Oncorhynchus mykiss W. 1792) Aeromonas hydrophila ve Yersinia ruckeri İzolasyonu ve Patolojik İncelemeler. Atatürk Üniversitesi Veteriner Bilimleri Dergisi, 1(1-2), 6-10.
• Sakai, M. (1999). Current status of fish immunostimulants. Aquaculture, 172, 63-92.
• Sarkodie, E.K., Zhou, S., & Chu, W. (2019). N-Acylhomoserine Lactones (AHLs), QseB/C Gene Detection, Virulence Factors and Antibiotics Resistance of Aeromonas hydrophila. Advances in Microbiology, 9(05), 495.
• Sorum, H. (2006) Antimicrobial drug resistance in fish pathogens. In Antimicrobial Resistance in Bacteria of Animal Origin. Aarestrup, F.M. (ed.). Washington, DC, USA: American Society for Microbiology Press, pp. 213–238 (Chapter 13).
• Theguardian, (2020). Antibiotics Powerful antibiotic discovered using machine learning for first time, https://www.theguardian.com/society/2020/feb/20/antibiotic-that-kills-drug-resistant-bacteria-discovered-through-ai Son erişim tarihi: 01.08.2020.
• Thenmozhi, S., Rajeswari, P., Kumar, B. S., Saipriyanga, V., & Kalpana, M. (2014). Multi-drug Resistant Patterns of Biofilm Forming Aeromonas hydrophila from Urine Samples. International Journal of Pharmaceutical Sciences and Research, 5(7), 2908.
• Timur, M. (1983). An outbreak of disease of farmed eel (Anguilla anguilla) due to Aeromonas hydrophila in Turkey. Histopathological and bacteriological studies. Ankara Üniversitesi Veteriner Fakultesi Dergisi, 30(3), 361-367.
• Uttley, A.H., Collins, C.H., Naidoo, J., George, R.C. (1988). Vancomycin-resistant enterococci, The Lancet, 1, 57-58. Werner, S.B., & Rutherford, G.W. (1990). Aeromonas wound infections associated with outdoor activities California, California Department of Health Care Services, 39(20), 334-335.
• Witte, W. (2000). Selective pressure by antibiotic use in livestock. International Journal of Antimicrobial Agents, 16, S19–S24.
• Zhu, W., Zhou, S., & Chu, W. (2020). Comparative proteomic analysis of sensitive and multi-drug resistant Aeromonas hydrophila isolated from diseased fish. Microbial Pathogenesis, 139, 103930. https://doi.org/10.1016/j.micpath.2019.103930.