ABSTRAKIkan nila atau Oreochromis niloticus merupakan ikan konsumsi masyarakat di Indonesia. Kasus kematian massal ikan nila terjadi di beberapa lokasi budaya di Jawa, Lombok, dan Sumatera yang disebabkan oleh infeksi Orthomyxovirus, dan disebut sebagai Tilapia Lake Virus (TiLV). Tujuan penelitian ini adalah untuk mendeteksi adanya infeksi TiLV dengan metode semi-nested Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR) pada kasus kematian massal ikan nila. Lokasi pengambilan sampel di Desa Sigerongan Kecamatan Lingsar, Lombok, Nusa Tenggara Barat. Analisis deteksi RT-PCR menggunakan sampel organ otak, ginjal, limpa, dan hati, selanjutnya dilakukan sekuensing. Hasil pengamatan terhadap gejala klinis terhadap ikan nila moribund terlihat kondisi mata yang buram/katarak, serta cekung, abrasi kulit, serta perubahan warna tubuh menjadi lebih gelap. Hasil analisis RT-PCR menunjukkan bahwa kejadian kematian massal pada ikan nila suspektif diakibatkan oleh infeksi RNA virus TiLV. Analisis sekuensing menunjukkan bahwa TiLV dari sampel ikan nila di Lombok mempunyai kesamaan identitas genetik 97% dengan TiLV asal Israel (Genebank Accession Number KU 751816.1). KATA KUNCI: nila; TiLV; seminested RT-PCR ABSTRACT:Case study of tilapia lake virus (TiLV) infection on nila (Oreochromis niloticus

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ABSTRAKTujuan dari penelitian ini adalah untuk membuat kultur sel primer dari sirip ekor ikan mas (Cyprinus carpio) dan diberi nama common carp tail (CCT). Explant ditumbuhkan dalam cawan kultur (culture flask) ukuran 25 cm 2 yang berisi media Leibovitz's L-15 dengan penambahan serum 20%, Penicillin 250 IU, Streptomycin 250 μg/mL, Kanamycin Sulfate 250 μg/mL dan L-Glutamin 2 mM, serta diinkubasi pada suhu 28 o C. Perbedaan perlakuan berupa waktu pergantian media dan konsentrasi media dilakukan untuk mendapatkan kultur sel primer. Hasil pengamatan menunjukkan bahwa explant menunjukkan pertumbuhan sel setelah diinkubasi selama 24 jam. Pembentukan sel selapis (monolayer) mulai terlihat pada hari ke-4. Pasase pertama dilakukan pada hari ke-21 saat konfluensi mencapai 65%. Pasase selanjutnya dilakukan setiap 3 minggu dimana konfluensi mencapai 70%-80%. CCT terdiri atas sel berbentuk fibroblas dan epitel, dan berhasil dipasase sebanyak 12 kali selama lebih 2 tahun pemeliharaan.KATA KUNCI: ikan mas, kultur sel primer, common carp tail (CCT) ABSTRACT: Development of primary cell culture from caudal fin of common carp (Cyprinus carpio). By: Tuti Sumiati, Lila Gardenia, and Agus Sunarto The objectives of this research were to develop primary cell from caudal fin of common carp (Cyprinus carpio) and designate it as Common Carp Tail (CCT

Penelitian ini bertujuan untuk mengetahui efikasi vaksin “trivalen” merupakan kombinasi dari tiga jenis antigen bakteri, yaitu: Aeromonas hydrophila-AHL0905-2, Streptococcus agalactiae-N14G, dan Mycobacterium fortuitum-31 untuk pencegahan penyakit bakteri potensial pada budidaya ikan air tawar. Ikan uji menggunakan ikan lele, nila, dan gurami; di mana masing-masing jenis ikan tersebut merupakan representasi dari jenis ikan yang rentan terhadap infeksi bakteri Aeromonas hydrophila, Streptococcus agalactiae, dan Mycobacterium fortuitum. Perlakuan yang diterapkan adalah formulasi sediaan vaksin kombinasi “trivalen”, dengan komposisi sebagai berikut: (A) formulasi proporsional yang mengandung ketiga jenis antigen dengan perbandingan 1:1:1 (v/v), (B) formulasi non-proporsional yang mengandung ketiga jenis antigen dengan perbadingan 1:3:3 (v/v), dan (C) tanpa pemberian vaksin sebagai kontrol. Vaksinasi diberikan melalui perendaman dalam larutan vaksin “trivalen” pada konsentrasi bakteri 107 cfu/mL selama 30 menit. Efikasi vaksin dievaluasi berdasarkan sintasan pada akhir uji tantang terhadap bakteri patogen target, dan selanjutnya dihitung nilai relative percentage survival (RPS). Hasil penelitian menunjukkan bahwa nilai RPS vaksin “trivalen” dengan kombinasi proporsional pada ikan lele terhadap bakteri A. hydrophila-AHL0905-2, ikan nila terhadap bakteri S. agalactiae-N14G, ikan gurami terhadap bakteri M. fortuitum-31 dan A. hydrophila-AHL0905-2 masing-masing sebesar 44,61%; 43,30%; 17,86%; dan 45,00%. Nilai RPS vaksin kombinasi “trivalen” non-proporsional pada ikan lele terhadap bakteri A. hydrophila-AHL0905-2, ikan nila terhadap bakteri S. agalactiae-N14G, ikan gurami terhadap bakteri M. fortuitum-31 dan A. hydrophila-AHL0905-2 masing-masing sebesar 39,61%; 40,00%; 7,14%; dan 45,00%. Vaksin “trivalen” berpotensi sebagai sediaan vaksin yang dapat digunakan untuk pencegahan penyakit pada budidaya air tawar yang disebabkan oleh ko-infeksi lebih dari satu jenis bakteri patogen.The study aimed to determine the efficacious of combined “three-valent” vaccine composed of three bacterial antigen for the prevention of bacterial diseases on freshwater aquaculture has been carried out at laboratory level. The study used catfish, tilapia, and giant goramy; that are representative of susceptible species against Aeromonas hydrophila infection, Streptococcus agalactiae infection, and Mycobacterium fortuitum infection. The treatments applied in this study included: (A). Proporsional formula of combined “three-valent” vaccine composed of three bacterial antigens at the ratio of 1:1:1 (v/v), (B). Non-proportional formula composed of three bacterial antigens at the ratio of 1:3:3 (v/v), and (C). Un-vaccinated group as a control. The vaccine was applied through immersion at the dose of 107 cfu/mL vaccine solution for 30 minutes. Vaccine efficacious was evaluated based on survival rate after challenge test against targetted pathogens, and then the relative percentage survival (RPS) was calculated. The study results revealed that the RPS values of proportional combined vaccine on catfish against A. hydrophila-AHL0905-2, on tilapia against S. agalactiae-N14G, on goramy against M. fortuitum-31 and A. hydrophila-AHL0905-2 were 44.61%; 43.30%, 17.86%, and 45.00% respectively. RPS values of non-proportional combined vaccine on catfish against A. hydrophila-AHL0905-2, on tilapia against S. agalactiae-N14G, on goramy against M. fortuitum-31 and A. hydrophila-AHL0905-2 are 39.61%; 40.00%, 7.14%, and 45.00% respectively. “Trivalent” vaccine is a promising combined-fish vaccine which can be used for prevention of bacterial fish diseases caused by co-infection of more than one type of pathogenic bacteria.

Streptococcosis is one of bacterial diseases in the culture of Tilapia, Oreochromis niloticus and has caused significant economic losses. Streptococcus iniae, is known as pathogen to marine and freshwater fishes whereas Streptococcus agalactiae is known as pathogen to Tilapia. The isolation and characterization of four isolates of S. agalactiae, were described from an infected Tilapia from Cirata Reservoir, West Java, in July 2008. Conventional and rapid identification systems were used to determine isolates of S. agalactiae from brain and kidney tissues. In this paper, we have characterized S. agalactiae and this was the first isolation of this bacteria from fish. The isolates were gram positive, catalase-negative, oxidase-negative,  haemolytic cocci colonies on blood agar. All of the of isolates were biochemically characterized with the API 20 Strep System (bioMerieux). Bacterial chromosomal DNA used in PCR assay was extr acted by heating method. The forward primer is Sdi 61: 5'-AGGAAACCTGCCATTTGCG-3' and the r ever se pri mer is Sdi 252: 5'-CAATCTATTTCTAGATCGTGG-3' with gene target 16S intergenic spacer and it has 192 bp in length. These primers were designed by Alpha DNA (Montreal, Quebec). The biochemical patterns of four isolates were rather different although almost all traits were similar with the exception of pyroglutamic acid (pyra) and L-arginin (ADH), for which we observed negative and positive reaction in this study. Therefore, some of the biochemical characteristics of the four isolates did not fit 100% with the typical patterns of S. agalactiae. However, the PCR result showed that this PCR assay is an effective tool for rapid and specific detection of S. agalactiae, the main pathogens involved in warm-water streptococcosis, obtained from pure culture of naturally infected fish. Therefore, it could be a useful alternative for culture-based routine diagnosis of warm-water streptococcal infections in fish.