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"ABSTRAKPenelitian kombinasi konsentrasi kuning telur sebagai krioprotektan belum pernahdilakukan pada ikan Botia (Chromobotia macrachanthus). Penelitian menggunakankonsentrasi kuning telur sebagai krioprotektan ekstraseluler dan metanol 10% sebagaikrioprotektan intraseluler. Konsentrasi kuning telur (0%, 5%, 7%, 9%, 11%, 13%, 15%,17%) dan penggunaan Carboxymethyl Cellulose (CMC) 1% terhadap motilitas,viabilitas dan abnormalitas spermatozoa ikan botia 24 jam pascakriopreservasi.Preservasi dilakukan pada tabung nitrogen cair dengan suhu -196°C. Berdasarkan hasiluji ANAVA satu arah menunjukkan pemberian berbagai konsentrasi kuning telurberpengaruh nyata (p<0,05) terhadap persentase motilitas, viabilitas, dan abnormalitasspermatozoa ikan Botia 24 jam pascakriopreservasi. Konsentrasi kuning telur optimumialah 15%, dengan nilai persentase motilitas (96,43 ± 1,49%), nilai persentase viabilitas(84,25 ± 1,26%) serta nilai persentase abnormalitas terendah (11,50 ± 1,29%). UjiTukey persentase nilai fertilitas telur 24 jam pascakriopreservasi tertinggi padakonsentrasi kuning telur 15% (50,64 ± 4,37%).

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ABSTRACTThe combination effect of egg yolk and 10% methanol on Botia fish spermatozoaquality and has not been performed, yet. Accordingly, the objrctive of study was: first,to evaluate the motility rate, viability rate, and abnormality of Botia fish spermatozoa 1day after cryopreservation. Second, to evaluate the fertility rate of Botia fish egg afterfertilized by cryopreserved sperm. The various concentration of egg yolk used were,5%, 7%, 9%, 11%, 13%, 15%, and 17% whereas the negative control (0%) used 10%methanol only without egg yolk. While, the positive of control used 1% of CMC. Botiasperm and egg were collected by hand stripping method. Physical and chemical ofbotia sperm had been observed by visual observation whereas the otility rate, vuiabilityrate, abnormality rate and fertility rate determined by light microscope. Botia fishsperm were mixed with cryoprotectand and extender before freezing at -196℃ (in LN).cryopreservation of botia fish sperm were conducted for one day. Based on one-wayANOVA test, gave the significant different between treatment group and control.Furthermore, according to Tuket test, they were gave the significant different (P<0.05)also among treatment group. Fifteen percent of egg yolk was optimum concentreationthat gave the highest motility rate, (96.43 ± 1.49%), and the highest viability (84.25 ±1.26%) and showed the lowest percentage of abnormality (11.50 ± 1.29%), and also thehighest fertility rate of Botia fish egg that (50,64 ± 4,37%) with protected by 15% ofegg yolk."

"Kriopreservasi sperma ikan botia Cromobotia macracanthus Bleeker, 1852 merupakan salah satu solusi untuk mengatasi kendala pemijahan induk botia yang bersifat musiman serta pematangan induk jantan dan induk betina yang tidak sinkron. Penelitian ini menggunakan krioprotektan metanol sebagai krioprotektan intraseluler dan madu sebagai krioprotektan ekstraselululer. Metanol umum digunakan sebagai krioprotektan intraseluler karena memiliki ukuran molekul yang kecil sehingga dapat menembus membran sel dengan mudah dan juga memiliki toksisitas yang lebih rendah dibandingkan dimetil sulfoksida DMSO dan dimethyl acetamide DMA . Madu merupakan bahan alami yang dapat digunakan sebagai krioprotektan ekstraseluler karena bersifat melindungi sel sperma dari luar sel. Tujuan penelitian yaitu untuk mengevaluasi pengaruh metanol dan madu sebagai krioprotektan terhadap kualitas serta kemampuan fertilisasi sperma ikan botia 48 jam pascakriopreservasi. Krioprotektan yang digunakan dalam penelitian yaitu metanol 10 dan madu dalam berbagai konsentrasi 0 ; 0,1 ; 0,3 ; 0,5 ; 0,7 dan 0,9. Rasio sperma dan larutan pengencer yang dalam penelitian ini yaitu 1:9. Ekstender yang digunakan yaitu larutan fish Ringer. Sperma disimpan dalam deep freezer -80o C selama 48 jam. Sperma segar dievaluasi terlebih dahulu untuk menguji kelayakan sperma untuk dikriopreservasi. Sperma segar dievaluasi secara makroskopis warna, bau, volume sperma, pH, secara mikroskopis motilitas, viabilitas dan abnormalitas, dan juga kemampuan fertilisasinya dengan menghitung persentase fertilisasi. Sperma pascakriopreservasi dievaluasi secara mikroskopis dan kemampuan fertilisasinya. Hasil penelitian menunjukkan bahwa krioprotektan metanol 10 dan berbagai konsentrasi madu tidak berpengaruh nyata P>0,05 terhadap motilitas dan abnormalitas spermatozoa ikan botia pascakriopreservasi, akan tetapi berpengaruh nyata terhadap viabilitas dan kemampuan fertilisasinya P.

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Cryopreservation of botia Cromobotia macracanthus Bleeker, 1852 sperm is one of the solutions to overcome the constraints of seasonal spawning botia spawning and maturation of male broodstock and female broodstock that is not synchronized. This study used cryoprotectant methanol as intracellular cryoprotectant and honey as extracellular cryoprotectant. Methanol is commonly used as intracellular cryoprotectants because its small molecular size so that can penetrate to cell membranes easily and also has lower toxicity than dimethyl sulfoxide DMSO and dimethyl acetamide DMA . Honey is a natural substance that can be used as an extracellular cryoprotectant that protects sperm cells from outside the cell.The objective of the study was to evaluate the effect of methanol and honey as a cryoprotectant on the quality and fertilization ability of 48 hours postcryopreservation botia sperm. Cryoprotectants used in the study were 10 methanol and honey in various concentrations 0, 0.1, 0.3, 0.5, 0.7 and 0.9. The ratio of sperm and diluent solution in this study is 1 9. Extender used is fish Ringer solution. Sperm is stored in deep freezer 80o C for 48 hours. Fresh sperm are evaluated to test the sperm feasibility for cryopreservation. Fresh sperm are evaluated macroscopically color, odor, sperm volume, pH, microscopically motility, viability and abnormality, as well as their fertilization ability by calculating the percentage of fertilization. Postcryoservation sperm is evaluated microscopically and its fertilization ability. The results showed that 10 methanol and various honey concentration as cryoprotectant had no significant effect P 0.05 on the motility and abnormalities of botia cryopreserved sperm, but it had significant effect on viability and fertilization ability P."