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RESEARCH ARTICLE
Morfologi, fisiologi, preservasi sel sperma ikan betok, Anabas testudineus Bloch 1792 dan ketahanannya terhadap kejut listrik
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Abstract
Good sperm quality and inactive condition during the electroporation process determines the success of transgenesis. The study aims to observe the morphology and physiology of sperm, and evaluate the effects of different voltage and number of electric shocks on motility and viability of climbing perch sperm. Observation of morphological characters by microscope with a magnification of 400X, physiological character with various parameters, such as the level of os-molarity, volume, pH, density and total of sperm. The climbing perch sperm consisted of three parts, namely the ovoid head, middle, and tail. Sperm viability in the water was 116±34 seconds. Sperm quantity, volume of semen, pH, and sperm density were increased after ovaprim injection (p<0.05). Osmolarity levels of body and testes of climbing perch ranged at 390-405 mOsmol kg-1. Sperm diluted in 13 g L-1 NaCl solution could be kept for a maximum of 60 minutes at room temperature (26°C) and cold temperature (4°C), while without NaCl solution could be 360 minutes. Fertilization and hatching rates of eegs fertilized by sperm form the minced testes, and larval survival up to 10 days post-hatching were similar with semi-natural spawning. Electroporation of climbing perch sperm diluted in 13 g/L NaCl solution was successfully performed using a program with a long shock of 0.5 milliseconds and the interval shock of 0.1 seconds. Increasing the voltage and number of shock decreased motility and viability of sperm (p<0.05). Those electric shock parameters can potentially be used to generate climbing perch transgenic by electroporation method.
Abstrak
Kualitas sperma yang baik dan dalam keadaan tak aktif selama proses elektroporasi sangat menentukan keberhasilan transgenesis. Penelitian ini bertujuan untuk mengeksplorasi karakter morfologis dan fisiologis sel sperma, serta menge-valuasi efek voltase dan jumlah kejut listrik berbeda terhadap motilitas dan viabilitas sperma ikan betok. Pengamatan karakter morfologi dilakukan pada mikroskop dengan pembesaran 400x, karakter fisiologi dengan berbagai parameter yakni tingkat osmolaritas, volume, pH, kepadatan dan total sperma. Hasil penelitian menunjukkan bahwa sperma ikan betok terdiri atas tiga bagian, yakni kepala yang berbentuk oval, bagian tengah, dan ekor sperma. Viabilitas sperma ikan betok dalam air adalah selama 116±34 detik. Induksi ovaprim dapat meningkatkan volume sperma, pH, kepadatan dan total sperma (p<0,05). Tingkat osmolaritas tubuh dan testis ikan betok (390-405 mOsmol kg-1). Penyimpanan sperma yang diencerkan dengan larutan NaCl 1,3% hanya dapat bertahan maksimum selama 60 menit pada suhu ruang (26°C) dan suhu dingin (4°C), sedangkan tanpa pengenceran dapat bertahan selama 360 menit. Derajat pembuahan dan kelangsungan hidup larva hingga 10 hari pascatetas menggunakan sperma dari testes cacah sama dengan pemijahan semi alami. Elektroporasi sperma ikan betok dengan pengencer larutan NaCl 1,3% berhasil dilakukan menggunakan lama kejut 0,5 milidetik, dan jeda antar kejut 0,1 detik. Peningkatan voltase dan jumlah kejut yang diberikan saling berinter-aksi menurunkan motilitas dan viabilitas sperma (p<0,05). Metode kejut listrik tersebut berpotensi digunakan untuk menghasilkan ikan betok transgenik dengan metode elektroporasi.
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