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All Journal Saintek Perikanan : Indonesian Journal of Fisheries Science and Technology JFMR (Journal of Fisheries and Marine Research) Depik Jurnal Ilmu-Ilmu Perairan, Pesisir dan Perikanan Depik Jurnal Ilmu-Ilmu Perairan, Pesisir, dan Perikanan
Herlina, Cucun
Brawijaya University
Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Earth & Planetary Sciences Education Engineering Environmental Science

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UJI IN VITRO DAN KARAKTER PROTEIN EKSTRASELULER (ECP) Edwardsiella tarda DENGAN KONSENTRASI ETHANOL BERBEDA (In Vitro Test and Character of Extracellular Protein (ECP) Edwardsiella tarda with Different Ethanol Concentration) Cucun Herlina; Uun Yanuhar; Maftuch Maftuch
Saintek Perikanan : Indonesian Journal of Fisheries Science and Technology Vol 15, No 2 (2019): SAINTEK PERIKANAN
Publisher : Fakultas Perikanan dan Ilmu Kelautan, Universitas Diponegoro

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (732.923 KB) | DOI: 10.14710/ijfst.15.2.105-111

Abstract

Protein ekstraseluler (ECP) bakteri memiliki kemampuan imunogenik dan dapat meningkatkan sistem imun tubuh inang. Komponen ECP diantaranya flagellin, protease dan metalloprotease mampu berdifusi pada sel inang untuk mengaktifkan respon imun. Tujuan penelitian adalah untuk mengetahui produksi protein ekstraseluler E. tarda dengan presipitasi ethanol pada konsentrasi berbeda sebagai potensi protein imunogenik secara in vitro. Metode penelitian yaitu kultur bakteri E. tarda, preparasi protein ekstraseluler, presipitasi protein, konsentrasi protein ditentukan denganspektrofotometer dan SDS-PAGE, uji in vitro protein ECP E. tarda, dan uji nilai RPS. Hasil protein ekstraseluler yang diperoleh dengan presipitasi ethanol tertinggi pada konsentrasi 90% sebanyak 5,69 mg/ml dan terendah pada konsentrasi 80% sebanyak 4,81 mg/ml. Hasil karakterisasi protein dengan SDS-SDSPAGE antara 30-60 kDa. Hasil uji in vitro vaksin E. tarda (presipitasi ethanol 100%) baik dari uji viabilitas maupun sterilitas menunjukkan tidak terdapat koloni yang tumbuh. Hasil nilai RPS pada ikan nila (Oreochromis niloticus) mencapai 100% dengan masa pemelihaaraan 9 hari pasca imunisasi. ECP bacteria have immunogenic abilities and can increase the host's immune system. ECP components which are flagellin, protease, metalloprotease can diffuse to host cell for the activation immune response. The study aimed to determine the production of extracellular protein E. tarda with ethanol precipitation at different concentrations as a potential immunogenic protein by in vitro test. The research methods were E. tarda bacterial culture, extracellular protein preparation, protein precipitation, protein concentration determined by spectrophotometer and SDS-PAGE, in vitro ECP E. tarda protein test, and RPS value test. The results of extracellular protein obtained by the highest ethanol precipitation at a concentration of 90% as much as 5.69 mg/ml and the lowest at 80% concentration as much as 4.81 mg/ml. The results of the characterization of proteins with SDS-SDS-PAGE between 30-60 kDa. The results of the in vitro test of E. tarda vaccine (100% ethanol precipitation) from both the viability and sterility tests showed that there were no growing colonies. The results of the RPS in tilapia (Oreochromis niloticus) reach 100% with a maintenance period of 9 days after immunization.
Uji Viabilitas Kandidat Vaksin Bakteri Aeromonas Hydrophila Menggunakan Metode Formalin-Killed dengan Konsentrasi Berbeda: Viability Assay of Aeromonas Hydrophila as Candidate Vaccine Using Formalin-Killed Method at Different Concentration Herlina, Cucun; Maftuch, Maftuch; Fadjar, Mohamad
JFMR (Journal of Fisheries and Marine Research) Vol. 9 No. 1 (2025): JFMR on March
Publisher : Faculty of Fisheries and Marine Science, Brawijaya University, Malang, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21776/ub.jfmr.2025.009.01.7

Abstract

Salah satu upaya pencegahan yang dapat dilakukan untuk mengatasi penyakit pada ikan yaitu menumbuhkan kekebalan seperti vaksinasi karena kekebalan tubuh yang meningkat pada ikan dapat mencegah infeksi penyakit. upaya telah dilakukan untuk mengembangkan program imunisasi yang efektif dengan menggunakan vaksin yang dibuat dari sel utuh bakteri yang diinaktivasi menggunakan formalin dan/atau pemanasan Tujuan penelitian ini untuk mengetahui hasil uji viabilitas dan hasil uji adhesi sel epitel usus menggunakan vaksin inaktif formalin dengan konsentrasi berbeda. Metode dalam penelitian ini adalah metode eksperimen dengan rancangan percobaan menggunakan rancangan acak lengkap (RAL) terdiri dari 3 perlakuan (A=2%, B=3%, dan C=4%) dan kontrol dengan 3 kali ulangan. Hasil penelitian diperoleh hasil kandidat vaksin inaktif bakteri A.hydrophila sebanyak 9 ml/tabung falcon. Infeksi bakteri A. hydrophila dengan kepadatan 109 cfu/ml sebanyak 0,1 ml dengan masa pemeliharaan 1 minggu ditandai terjadi pengerasan di bagian perut dan terdapat kerusakan pada sirip punggung pada hari ketujuh. Hasil uji viabilitas menunjukkan tidak terdapat pertumbuhan koloni bakteri A. hydrophila dibandingkan dengan kontrol. Hasil uji adhesi sel epitel usus pada kontrol dan perlakuan A, B, C diperoleh rata-rata 10.3, 0.73, 0.43, dan 0.23 bakteri/10 sel epitel. Analisis uji ANOVA diperoleh nilai uji F Hitung > F Tabel, artinya perlakuan memberikan pengaruh sangat berbeda signifikan antar semua perlakuan. Hasil analisa regresi diperoleh (R2) sebesar 0.66 bahwa hubungan keterkaitan antara vaksin inaktif konsentrasi formalin berbeda dengan jumlah bakteri A. hydrophila yang menempel pada sel epitel sebesar 66%.   One preventive effort to tackle diseases in fish is by boosting immunity, such as through vaccination, as increased immunity in fish can prevent disease infections. Efforts have been made to develop effective immunization programs using vaccines made from whole bacterial cells inactivated using formalin and/or heat. The objective of this study was to determine the results of viability tests and intestinal epithelial cell adhesion tests using formalin-inactivated vaccines with different concentrations. The method used in this study was an experimental approach with a completely randomized design (CRD) consisting of three treatments (A=2%, B=3%, and C=4%) and a control, each with three replications. The study results showed that the candidate inactivated vaccine of A. hydrophila bacteria was obtained at 9 ml/falcon tube. A. hydrophila bacterial infection with a density of 10⁹ CFU/ml at 0.1 ml and a maintenance period of one week was characterized by abdominal hardening and dorsal fin damage on the seventh day. The viability test results showed no bacterial colony growth of A. hydrophila compared to the control. The intestinal epithelial cell adhesion test results for the control and treatments A, B, and C showed average bacterial adhesion of 10.3, 0.73, 0.43, and 0.23 bacteria/10 epithelial cells, respectively. ANOVA test analysis showed that the calculated F-value was greater than the table F-value, indicating a highly significant effect among all treatments. Regression analysis resulted in an R² value of 0.66, suggesting that 66% of the variation in the adhesion of A. hydrophila bacteria to epithelial cells was associated with different formalin concentrations in the inactivated vaccine.
Effect of Extracellular Crude Protein (ECP) on expression IL-1, IL-6, IFN- in nile tilapia (Oreochromis niloticus) Herlina, Cucun; Fadjar, Mohammad; Maftuch, Maftuch; Yanuhar, Uun
Depik Jurnal Ilmu Ilmu Perairan, Pesisir, dan Perikanan Vol 14, No 3 (2025): SEPTEMBER 2025
Publisher : Faculty of Marine and Fisheries, Universitas Syiah Kuala

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.13170/depik.14.3.43338

Abstract

Modern strategies incorporate preventive measures such as extracellular products, as soluble proteins with antigenic properties, as well as generating a broad immune response. The aim of research is to determine and analyze the effect of the extracellular product of E. tarda on enhancing the immunity of Nile tilapia (O. niloticus). The research design with treatment durations of d0, 3rd, 6th, 9th day, and control (A, B, C, D, K). ECP vaccination was injected intraperitoneally at a dose of 0.1 ml/fish. The parameters analyzed are the expression of IL-1, IL-6, and IFN-, phagocytic activity, leukocyte differential, total white blood cells, and relative percent survival. The results of phagocytosis activity were highest in treatment C at 84.18% 1.81. The highest leukocyte differential in treatment C was lymphocytes at 56% 2.00, monocytes at 28% 2.00, and 16% 0.00, with a total leukocyte count of 29 x 10^3 cells 2.64, and RPS values ranging from 83.33% to 100% over 9 days of post-vaccination maintenance. Then, post-vaccination, there was an increase in the expression levels of the cytokine genes IL-1 and IL-6, but there was no increase in the expression levels of the IFN- gene compared to the control in Nile tilapia. (O. niloticus).Keywords:ECPIL-1IL-6IFN-Immune Response
Effect of Extracellular Crude Protein (ECP) on expression IL-1, IL-6, IFN- in nile tilapia (Oreochromis niloticus) Herlina, Cucun; Fadjar, Mohammad; Maftuch, Maftuch; Yanuhar, Uun
Depik Jurnal Ilmu Ilmu Perairan, Pesisir, dan Perikanan Vol 14, No 3 (2025): SEPTEMBER 2025
Publisher : Faculty of Marine and Fisheries, Universitas Syiah Kuala

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.13170/depik.14.3.43338

Abstract

Modern strategies incorporate preventive measures such as extracellular products, as soluble proteins with antigenic properties, as well as generating a broad immune response. The aim of research is to determine and analyze the effect of the extracellular product of E. tarda on enhancing the immunity of Nile tilapia (O. niloticus). The research design with treatment durations of d0, 3rd, 6th, 9th day, and control (A, B, C, D, K). ECP vaccination was injected intraperitoneally at a dose of 0.1 ml/fish. The parameters analyzed are the expression of IL-1, IL-6, and IFN-, phagocytic activity, leukocyte differential, total white blood cells, and relative percent survival. The results of phagocytosis activity were highest in treatment C at 84.18% 1.81. The highest leukocyte differential in treatment C was lymphocytes at 56% 2.00, monocytes at 28% 2.00, and 16% 0.00, with a total leukocyte count of 29 x 10^3 cells 2.64, and RPS values ranging from 83.33% to 100% over 9 days of post-vaccination maintenance. Then, post-vaccination, there was an increase in the expression levels of the cytokine genes IL-1 and IL-6, but there was no increase in the expression levels of the IFN- gene compared to the control in Nile tilapia. (O. niloticus).Keywords:ECPIL-1IL-6IFN-Immune Response
Co-Authors Fadjar, Mohammad Maftuch Maftuch MOHAMAD FADJAR Yanuhar, Uun