Isolasi, Purifikasi dan Immunogenitas Protein Outer Membran Vibrio Alginolyticus pada Ikan Kerapu Macan (Epinephelus fuscoguttatus)

https://doi.org/10.22146/jfs.57

Desrina Desrina(1*), Arief Taslihan(2), Ambariyanto Ambariyanto(3), Ervia Yudiati(4), Yulius Docang Casessar(5), Raden Bagus Sugio Sumanta(6), Triyanto Triyanto(7), Hotnida Junita Situmeang(8), Langkah Sembiring(9)

(1) Program Studi Budidaya Perairan, Jurusan Perikanan Fakultas Perikanan dan Ilmu Kelautan UNDIP
(2) Balai Besar Pengembangan Budidaya Air Payau
(3) Jurusan Kelautan Fakultas Perikanan dan Ilmu Kelautan UNDIP
(4) Jurusan Kelautan Fakultas Perikanan dan Ilmu Kelautan UNDIP
(5) Jurusan Kelautan Fakultas Perikanan dan Ilmu Kelautan UNDIP
(6) Jurusan Kelautan Fakultas Perikanan dan Ilmu Kelautan UNDIP
(7) Jurusan Perikanan Fakultas Pertanian UGM
(8) Jurusan Biologi Fakultas Biologi UGM
(9) Jurusan Biologi Fakultas Biologi UGM
(*) Corresponding Author

Abstract


The main objectives of this research were to isolate, purify and determine the immunogenicity of the outer membrane protein of V. alginolyticus. The outer membrane protein was isolated by sonication, sodium dodecyl sulphate-polyacrylamide agarose gel electrophoresis (SDS-PAGE) and purified by electroelusion method. Four outer membrane proteins were obtained: namely 32.0; 37.83; 64.13 and 73.43 kDa. Its immunogenicity in grouper Epinephelus fuscoguttatus was compared to that of bacterin of the same isolate. The immunogenicity test was conducted by intra peritoneal injection method. Each protein was dissolved in sterile Phospate Buffer Saline (PBS) and Freund’s Complete Adjuvant (FCA) (1:1) and injected 5 µg/ fish (fish weight 10-15 g). Bacterin (106, 107 and 108 cells/ml) was prepared in the same manner and injected at dosage of 0,2 ml/fish. Control fish were injected with 0.2 ml sterile PBS (pH 7.2). Booster was done a week later by injecting protein or bacterin with the Freund’s Incomplete Adjuvant (FIA). The agglutination test of antibody produced recognized cell surface protein of the whole cell of V. alginolyticus. Outer membrane protein 73.43 kDa was more immunogenic than the rest of proteins and bacterin, based on agglutinating antibodi titer.

Keywords


Epinephelus fuscoguttatus, immunogenicity, outer membrane protein, Vibrio alginolyticus

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References

Ahn, S., J. Han, J. Lee, K. Park, and I. Kong. 2005. Identification of an iron regulated hemin-binding outer membrane protein Hup O in Vibrio fluvialis: Effects on hemolytic activity and the oxidative stress response. Infection and Immunity. 73 (2): 722-729.

Barrow, G.I., and R.K.A. Feltham. 1993. Cowan and Steel’s manual for the identification of medical bacteria. Cambridge University Press. Cambridge. 329 p.

Baumann, P., A.L. Furniss, and J.V. Lee. 1984. Genus Vibrio. In: Bergey’s Manual of Systematic Bacteriology Vol. 1. N.R. Krieg and J.G. Holt (Eds.). William and Wilkins, Baltimore. Maryland : 528-550.

Bondad-Reantaso, M.G., S. Kanchana khan and S. Chinabut. 2000. Review of grouper diseases and health management strategies for grouper and other marine finfish diseases. Regional Workshop on Sustainable Seafarming and Grouper Aquaculture. Collaborate APEC Grouper Research and Development Network. FWG 01/99. 17-20 April 2000. Medan. Indonesia. 232 p.

Casadevall, A., and L. Pirofski. 1999. Host-pathogen interaction: redifining the basic concepts of virulence and pathogenicity. Infection and Immunity. 67 (8): 3703-3713.

Colquhoun, D.J. and H. Sorum. 1998. Outer membrane protein expression during in vivo cultivation of Vibrio salmonicida. Fish and Shellfish Immunology. 8: 367-377.

Davey, M.L., R.E.W. Hancock, and L.M. Mutharia. 1998. Influence of culture conditions on expression of the 40-kilodalton porin protein of Vibrio anguillarum serotype 02. Applied and Environmental Microbiology. 64(1): 138-146.

Deitsch, K.W., E.R. Moxon, and T.E. Wellems. 1997. Shared themes of antigenic variation and virulence in bacterial, protozoal and fungal infections. Microbiology and Molecular Biology Reviews. 61(3): 281-293.

Esteve-Gassent, M.D. and C. Amaro. 2004. Immunogenic antigens of the eel pathogen Vibrio vulnivicus serovar E. Fish and Shellfish Immunology. 17: 277-291.

Finlay, B. B. and S. Falkow. 1997. Common themes in microbial pathogenicity, revisited. Microbiology and Molecular Biology Reviews. 61 (2): 136-169.

Horne, M.T., and A.E. Ellis. 1988. Commercial production and licensing of vaccines for fish. In: Fish Vaccination. A.E.Ellis (Ed.). Academic Press Inc. Sandiego. CA.: 47-64

Kanchanakhan, S. 1996. Diseases of cultured grouper. AAHRI Newsletter Article. 5 (2): 1-4.

Kuby, J. 1992. Immunology. W.H Freeman and Company. New York. 585 p.

Neidhardt, F.C., J.L. Ingraham, and M. Schaechter. 1990. Physiology of the bacterial cell: a molecular approach. Sinauer Associates, Inc. Sunderland, Massachusetts. 506 p.

Nitimulyo, K.H., A. Isnansetyo, Triyanto, I. Istiqomah, dan M. Murdjani. 2005a. Isolasi , identifikasi dan karakterisasi Vibrio spp patogen penyebab vibriosis pada kerapu di Balai Budidaya Air Payau Situbondo. Jurnal Perikanan. VII (2): 80-94.

Nitimulyo, K.H., A. Isnanstyo, Triyanto, M. Murdjani, dan L. Solichah. 2005b. Effektifitas vaksin polivalen untuk pengendalian vibriosis pada kerapu tikus (Cromoliptes altivelis). Jurnal Perikanan. VII (2): 95-100.

Rahman, M.H. and K. Kawai. 2000. Outer membrane proteins of Aeromonas hydrophila induce protective immunity in goldfish. Fish and Shellfish Immunology. 10: 379-382.

Rahman, M.H., A. Kuroda, J.M. Dijkstra, I. Kiryu, T. Nakanishi, and M. Ototake. 2002. The outer membrane fraction of Flavobacterium psychrophilum induces protective immunity in rainbow trout and ayu. Fish and Shellfish Immunology. 12: 169-179.

Salati, F. 1988. Vaccination against Edwardsiella tarda. In: Fish vaccination. A.E. Ellis (Ed.). Academic Press. San Diego. CA: 135-151.

Shamsudin, M.N. 1994. Pathogenesis of Flexybacter columnaris and immunity in channel catfish. Dissertation. Departement of Fisheries and Allied Aquaculture. Auburn University, Auburn, Alabama. 207 p.

Stoskopf, M.K. 1993. Bacterial diseases of marine tropical fish. In: Fish medicine. M.K. Stoskopf (Ed.). W.B. Saunders Company. Philadelphia, PA.: 635-639.

Sprott, G.D., S.F. Koval, and C.A. Schnaitman. 1994. Cell fractionation. In: Methods for general and molecular microbiology. P. Gerhardt, R.G.E. Murray, W. A. Wood and N.R. Krieg (Eds.). American Society for Microbiology. Washington D.C. 72-103

Taslihan, A., M. Murdjani, C. Purbomartono, dan E. Kusnendar. 2000. Bakteri patogen penyebab penyakit mulut merah pada ikan kerapu tikus (Cromileptes altivelis). Jurnal Perikanan. II (2): 57-62.

Van derWounde, M.W., and A.J. Bäumler. 2004. Phase and antigenic variation in bacteria. Clinical Microbiology Review. 17 (3): 581-611.

Vinitnantharat, S., J.A. Plumb and A.E. Brown. 1993. Isolation and purification of an outer membrane protein of Edwarsiella ictaluri and its antigenicity to channel catfish (Ictalurus punctatus). Fish and Shellfish Immunology. 3: 401-409.



DOI: https://doi.org/10.22146/jfs.57

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Jurnal Perikanan Universitas Gadjah Mada (print ISSN 0853-6384; online ISSN 2502-5066) is published by Department of Fisheries, Universitas Gadjah Mada in collaboration with Semnaskan UGM (Seminar Nasional Tahunan Hasil Perikanan dan Kelautan) and ISMFR (International Symposium on Marine and Fisheries Research).

 

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