Immunodiagnosis Infeksi Aeromonas hydrophila  pada Ikan

Yuli Purwandari Kristianingrum; Sitarina Widyarini; Kurniasih Kurniasih; Bambang Sutrisno; Charles Rangga Tabbu; Sugiyono Sugiyono

doi:10.22146/jsv.38858

Immunodiagnosis Infeksi Aeromonas hydrophila pada Ikan

https://doi.org/10.22146/jsv.38858

Yuli Purwandari Kristianingrum^(1*), Sitarina Widyarini⁽²⁾, Kurniasih Kurniasih⁽³⁾, Bambang Sutrisno⁽⁴⁾, Charles Rangga Tabbu⁽⁵⁾, Sugiyono Sugiyono⁽⁶⁾

(1) Fakultas Kedokteran Hewan, Universitas Gadjah Mada, Yogyakarta
(2) Fakultas Kedokteran Hewan, Universitas Gadjah Mada, Yogyakarta
(3) Fakultas Kedokteran Hewan, Universitas Gadjah Mada, Yogyakarta
(4) Fakultas Kedokteran Hewan, Universitas Gadjah Mada, Yogyakarta
(5) Fakultas Kedokteran Hewan, Universitas Gadjah Mada, Yogyakarta
(6) Fakultas Kedokteran Hewan, Universitas Gadjah Mada, Yogyakarta
(*) Corresponding Author

Abstract

Aeromonas hydrophila causes a disease that often infects fish and is known as Motile Aeromonas Septicaemia (MAS), Hemorrhagi Septisemia, Ulcer disease or Red-Sore disease. The aims of this study were to develop polyclonal antibody of Aeromonas hydrophila in the rabbits to confirm the diagnosis of Aeromonas hydrophila in the fish by immunohistochemistry staining method. Preparation of polyclonal antibodies was performed on the rabbits used to Aeromonas hydrophila bacteria that have been tested biochemically by intravenous and intraperitoneal injection. Doses of Aeromonas hydrophila bacteria were 10⁹CPU/ml of 0.5 ml at first injection, 1 ml at second injection, 2 ml at thirth injection and 3 ml at fourth injection. Blood serum collection was performed at week 5 after injection from an ear and intracardial vein. The result of antibody titer was 2⁸ = 1024 which measured by tube test. Furthermore, polyclonal antibody was used to immunohistochemistry staining with 400x dilution. The results of the staining showed that an immunopositive reaction in the liver, skin,lien, gill, kidney, and heart of fish to Aeromonas hydrophila antibody. The research conclution was polyclonal antibody from rabbit can be used to accurately confirm the diagnosis of Aeromonas hydrophila based on antigen and antibody reaction.

Keywords

Aeromonas hydrophila; immunohistochemistry; antigen; antibody

Full Text:

PDF

References

Austin, B., Austin, D. A. (2007) Bacterial Fish Pathogens : Diseases of Farmed and Wild Fish. Fourth Edition. Springer Inc., New York. : 83-84.

Bootland, L.M., dan Leong, J.A. (1992) Staphylococcal Coaglutination, a Rapid Method of Identifying Infectious Hematopoetic Necrosis Virus. Appl. and Env. Microbiol. : 6-13

Buller, N. B. (2014) Bacteria and Fungi from Fish and Other Aquatic Animals : A Practical Identification Manual. Second Edition. CAB International Publishing, Oxfordshire. : 23, 151-152, 458

Cartwright, G.A., Chen, D., Hanna, P.J., Gudkovs, N., Tajima, K. (1994) Immunodiagnosis of virulent strains of Aeromonas hydrophila associated with epizootic ulcerative syndrome (EUS) using monoclonal antibody. J. Fish Disease. 17: 123-133

Coscelli, G.A., Bermudez, R., Sancho Silva, A.R., Ruiz de Ocenda, M.V., Quiroga, M.I. 2014. Granulomatous dermatitis in turbot (Schopthalmus maximus L.) associated with natural Aeromonas salmonicida subps.salmonicida infection. Aquaculture 428: 111-116

Coscelli, G.A., Bermudez, R., Losada, A.P., Failde, L.D., Santos, Y., Quiroga, M.I. (2014) Acut Aeromonas salmonicida infection (Schopthalmus maximus L.). Histopathological and immunohistochemical studies. Aquaculture 430: 79-85

Deen, A. E. N. E., Dorgham, S. M., Hassan, A. H. M., Hakim, A. S. (2014) Studies on Aeromonas hydrophila in Cultured Oreochromis niloticus at Kafr El Sheikh Governorate, Egypt with Reference to Histopathological Alterations in Some Vital Organs. World Journal of Fish and Marine Sciences 6(3): 233-240.

Dosim., Hardi, E. H., Agustina. (2006) Efek Penginjeksian Produk Intraseluler (ICP) dan Ekstraseluler (ECP) Bakteri Pseudomoas sp. terhadap Gambaran Darah Ikan Nila (Oreochromis niloticus). Jurnal Ilmu Perikanan Tropis, 19(1): 1-7.

FAO (Food and Agriculture Organization of the United Nations). (2009) FAO Yearbook : Fishery and AquacultureStatistics. http://www.fao.org/fishery/ (Diakses pada 7 Januari 2017).

Farto, R., Milton, D.I..,Bermudez, M., Nieto, T. (2011) Colonization of turbot tissues by virulent and avirulent Aeromonas salmonicida subsp. salmonicida leucocyte strains during infection. Dis.Aquat.Org. 95: 167-173

Irianto, A. (2006) Patologi Ikan Teleostei. Gadjah Mada University Press, Yogyakarta: 144-147.

Iskandar, R., Elrifadah. 2015. Pertumbuhan dan Efisiensi Pakan Ikan Nila (Oreochromis niloticus) yang Diberi Pakan Buatan Berbasis Kiambang. Jurnal Medika Veterinaria 40(1). 18-24.

Korbsrisate, S., Dumnin, S., Chawengkirttikul, R., Gherunpong, V., Eampokalap, B., Gongviseisoog, C., Janyapoon, K., Lertpocasombat, K., and Shimada, T. (2002) Distribution of Aeromonas hydrophila serogroups in different clinical samples and the development of polyclonal antibodies for rapid identification of the genus Aeromonas by direct agglutination. Microbial Immunol. 46: 875-879

Lamers C H J, De Hass M J M. (1985) Antigen localization in the lymphoid organs of carp (Cyprinus carpio) Cell Tissue Res. 242:491–498

Merino, S., Rubires, X., Aguilar, A., Tomas,, J.M. (1997) The role of flagella and motility in the adherence and invasion to fish cell lines by Aeromonas hydrophila serogroup O:34 strains. FEMS Microbiol. Lett. 151: 213-217

Moral C.H., Castillo E.F., Fierro P.L., Cortés A.V., Castillo J.A., Soriano, A.C., Salazar, M.S., Peralta, B.R., and Carrasco, G.N. (1998) Molecular Characterization of the Aeromonas hydrophila aroA Gene and Potential Use of an Auxotrophic aroA Mutant as a Live Attenuated Vaccine. Infect immun 66(5): 1813-1821

Oakey, H.J., Gibson, L.F. and George, A.M. 1999. DNA probe specific for Aeromonas hydropilla (HG1). J Appl. Microbiol., 86: 187-193

Rey, A., Verjan, N., Ferguson, H. W., Iregui, C. (2009) Pathogenesis of Aeromonas hydrophila Strain KJ99 Infection and its Extracellular Products in Two Species of Fish. The Veterinary Records 164(1): 493-499.

Sumithra, T.G., Chaturvedi, V.K., Grupta, P.K., Siju, S.J., Susan, C., Bincy, J.,Laxmi, U.,Sunita, S.C., Rai, A.K. (2014) Development of Simple Method for Rapid Identification of Organism causing Anthrax by Coagglutination Test. Biologicals 42: 316-321

Swan, L and White, M.R. (1989) A Guide to Approved Chemicals in Fish Production and Fishery Resource Management: Diagnosis and Tretment of Aeromonas hydrophila infection of Fish. Aqua Culture Extension. University of Arkansas Cooperative Extension Service.

Woo, P. T. K. (2006) Fish Diseases and Disorders : Protozoan and Metazoan. Volume 1. CABI Publishing, Canada : 163-165, 321-322.

Woo, P. T. K., Bruno, D. W. (2011) Fish Diseases and Disorders : Viral Bacterial and Fungal Infection. Volume 3. CABI Publishing, Canada: 424-455.

Yardimci,B., Aydin, Y. (2011) Pathological Findings of Experimental Aeromonas hydrophila Infection in Nile Tilapia (Oreochromis niloticus). Ankara Univ Vet Fak Derg 58(1): 47-54.

Yoshimizu, M. dan Kimura, T. (1985) A Coagglutinationtest with antibody sensitized Staphylococci for rapid and simple diagnosis of Bacterial and Viral Diseases of Fish. Fish Pathology, 20 : 243-261

Zhang, Y.L.,Ong, C.T., and Leung, K.Y. (2000) Molecular analysis of genetic differences between virulent and avirulent strains of Aeromonas hydropilla isolated from disease fish. Microbiol, 146: 999-1009

DOI: https://doi.org/10.22146/jsv.38858