HighlihgtDevelop primary cultures derived From tissue tails fins, gills, kidney and spleen from local Indonesian carp (Cyprinus carpio).Primary culture cell with L15 Mediacell cultures consist of two type Fibroblast-like and epithelial –like cell AbstractThe fish cell lines technology have been developed for the interests of the fisheries world. This study aimed at developing a primary cell line from gill, kidney, spleen, and caudal fin of a common carp (Cyprinus carpio). A healthy common carp weighing 20 g (~1 month) was collected from the Cijeruk Fish Seed Center, Bogor. The development of primary cell lines from the gill, fin, tail, kidney and spleen tissue was performed in cell culture medium Leibovitz’s L-15 supplemented with 20% serum fetal bovine, 250 IU Penicillin, 250 µg / ml kanamycin sulfate and 2Mm L-Glutamine, and incubated at 28°C. Primary cell lines of caudal fin and gill began to form a monolayer on day 17 after culture. While the development of cell lines from kidney and spleen, although the initiation of cells and cells spread on the surface into a monolayer, was not perfect; therefore, the passage was unable to be done. Microscopic observations and Giemsa staining showed primary cell lines of caudal fin and gill based on cell morphology consisted of two cell types, fibroblast-like cells and epithelial-like cells. The first passage was done on day 17 when the confluence was more than 50%. The next passage was carried out every 3 weeks when confluence reached 70% -80%. The primary cell culture of gill was successfully passaged as much as 72 and the caudal fin was successfully passed as much as 89 times over 7 years. These new cell lines can be further used to propagate fish viruses and other biotechnology assays.
Ikan kerapu dan kakap merupakan komoditas ikan penting di Indonesia yang memiliki berbagai jenis spesies maupun hasil silangannya. Salah satu permasalahan dalam kegiatan budidaya kelompok ikan ini yaitu adanya ancaman serangan virus Viral Necrosis Virus (VNN). Pada penelitian ini, dilakukan upaya pemetaan Betanodavirus sebagai penyebab VNN pada sentra budidaya ikan kerapu maupun kakap di wilayah Indonesia. Pemetaan genomik daerah RNA2 (coat protein) Betanodavirus dilakukan berdasarkan pada sejumlah 355 ekor ikan sampel dengan berbagai ukuran. Pengujian Betanodavirus dilakukan dengan metode Reverse Transcriptase – Nested PCR. Sedangkan untuk mengetahui sekuen dan hubungan kekerabatan lebih detail dilakukan sekuensing DNA dan analisa filogenetik. Hasil penelitian menunjukkan bahwa Betanodavirus yang ditemukan pada sampel yang diperoleh merupakan golongan dari Red-spotted Grouper Nervous Necrosis Virus (RGNNV). Selain itu, dari sejumlah sampel yang diperoleh dapat dikategorikan menjadi delapan sekuen utama. Berdasarkan analisa filogenetik yang telah dilakukan, Betanodavirus dari sampel ikan dapat digolongkan menjadi tiga kluster utama dengan tingkat kemiripan masing-masing kluster adalah 99.0%, 99.0% dan 96.02%.
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