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"Penelitian kriopreservasi spermatozoa ikan patin albino bertujuan untuk menganalisis ultrastruktur, fisiologi, dan molekuler spermatozoa ikan patin albino pasca kriopreservasi. Kriopreservasi dilakukan pada suhu -80°C selama 14 hari menggunakan kombinasi krioprotektan intraseluler yaitu metanol 10% dan krioprotektan ekstraseluler yaitu susu skim. Hasil ultrastruktur spermatozoa menunjukkan bahwa pada spermatozoa segar bagian membran sel kepala, mid piece, dan bagian flagel masih dalam kondisi utuh dan baik. Ultrastruktur spermatozoa pasca ekuilibrasi nampak ada perbesaran lebar dan panjang kepala spermatozoa dibandingkan spermatozoa segar, walaupun secara struktur masih tampak utuh. Ultrastruktur spermatozoa pasca pencairan tampak terjadi kerusakan membran bagian kepala dan flagel. Hasil pengukuran morfometri spermatozoa menunjukkan adanya peningkatan lebar kepala spermatozoa yaitu 1,59 µm pada spermatozoa segar menjadi 1,97 µm pada spermatozoa pasca ekuilibrasi dan 2,40 µm pada spermatozoa pasca pencairan. Demikian pula, terdapat perubahan panjang kepala spermatozoa yaitu 3,70 µm pada spermatozoa segar menjadi 3,81 µm pada spermatozoa pasca ekuilibrasi, dan 3,90 µm pada spermatozoa pasca pencairan. Analisis viabilitas spermatozoa didapatkan penurunan viabilitas spermatozoa pasca pencairan (61±2,30%) dibandingkan spermatozoa segar (92±0,58%) dan spermatozoa pasca ekuilibrasi (80±3,51%). Analisis fisiologi spermatozoa didapatkan penurunan fungsi mitokondria pada spermatozoa pasca ekuilibrasi (57±7%) dan spermatozoa pasca pencairan (42±3,2%) dibandingkan spermatozoa segar (98±2%). Analisis motilitas spermatozoa menunjukkan penurunan motilitas spermatozoa pasca ekuilibrasi (79±4,5%) dan spermatozoa pasca pencairan (30±3,2%) dibandingkan spermatozoa segar (87±1,5%). Penetasan telur pasca 24 jam fertilisasi pada perlakukan spermatozoa pasca ekuilibrasi didapatkan hasil lebih tinggi (64±17%) dibandingkan spermatozoa segar (38±4%), sedangkan spermatozoa pasca pencairan tidak ditemukan ada penetasan telur. Analisis molekular spermatozoa pada gen CO1 dan SOD2 didapatkan jumlah lesi gen SOD2 spermatozoa pasca ekuilibrasi yaitu 15,83 lesi / 10 kb dan spermatozoa pasca pencairan yaitu 17,14 lesi / 10 kb. Lesi gen CO1 pada spermatozoa pasca ekuilibrasi yaitu 9,24 lesi / 10 kb dan spermatozoa pasca pencairan yaitu 10,26 lesi / 10 kb. Sehingga disimpulkan kriopreservasi spermatozoa berpengaruh terhadap ultrastruktur, fisiologi, dan molekuler spermatozoa ikan patin albino.

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Research of cryopreservation on albino Pangasius catfish spermatozoa aims to analyze about ultrastructure, physiology, and molecular spermatozoa of albino Pangasius catfish post cryopreservation. Cryopreservation was carried out at -80°C for 14 days using a combination of intracellular cryoprotectants which is 10% methanol and extracellular cryoprotectant which is skim milk. The results of the spermatozoa ultrastructure showed that the cell membrane of the spermatozoa head, the midpiece, and the flagellum of fresh spermatozoa were still intact and good. The spermatozoa ultrastructure after post equilibration, shown enlargement of the head width and length compared to the fresh spermatozoa, although structurally were still intact. The ultrastructure of frozen-thawed spermatozoa, appeared a membrane damage at the head and flagellum. The results of spermatozoa morphometric measurements showed an increase at the head width of spermatozoa from 1.59 µm in fresh spermatozoa to 1.97 µm in post-equilibration spermatozoa and 2.40 µm in frozen-thawed spermatozoa. Similarly, there was an increase in the head length of spermatozoa, from 3.70 µm in fresh spermatozoa, to 3.81 µm in post-equilibration spermatozoa, and 3.90 µm in frozen-thawed spermatozoa. The viability analysis showed a decrease of frozen-thawed spermatozoa viability (61±2.30%) compared to fresh spermatozoa (92±0.58%) and post-equilibration spermatozoa (80±3.51%). The analysis physiology of spermatozoa showed a decrease in mitochondrial function in post equilibration spermatozoa (57±7%) and frozen-thawed spermatozoa (42±3.2%) compared to fresh spermatozoa (98±2%). The analysis of motility of spermatozoa showed a decrease in post equilibration spermatozoa (79±4.5%) and frozen-thawed spermatozoa (30±3.2%) compared to fresh spermatozoa (87±1.5%). Egg hatching after 24 hours of fertilization for the post-equilibration spermatozoa was higher (64±17%) than fresh spermatozoa (38±4%), whereas frozen-thawed spermatozoa were not hatched. The analysis of molecular on CO1 and SOD2 genes obtained the number of gene lesions in the spermatozoa SOD2 gene after equilibration were 15.83 lesions/10 kb and frozen-thawed were 17.14 lesions/10 kb. The CO1 gene lesions in post-equilibration spermatozoa were 9.24 lesions/10 kb, while the CO1 gene lesions in frozen-thawed spermatozoa were 10.26 lesions/10 kb. It can be concluded that there is an effect of cryopreservation on ultrastructure, physiology, and molecular in spermatozoa of albino Pangasius catfish."

"The effect of Dimethyl Formamide (DMF) as a cryoprotectant on spermatozoa quality of carp fish Cyprinus carpio race Majalaya 24 hours post-cryopreservation was studied. This study was conducted in the Environmental Engineering Laboratory. Puspiptek. Serpong. Sperm was collected by the hand stripping method and were immediately diluted with extender [6J and cryoprotectant (DMF). The final concentrations of DMF were 0. 3.75. 7.5. and ! 1.25%. respectively. Sperm were frozen in liquid nitrogen for 24 hours. All sperm were re-examined after thawing for motility. viability, and abnormality. The highest percentage of the molility and the viability of preserved sperm was showed or the DMF 7.5%. On the oiher hand the lowest percentage of the abnormality of preserved sperm was demonstrated by the DMF 7.5%. also. Accordingly. DMF 7.5% is the optimum levels that could preserved spermatozoa still in a good quality 24 hours posl-cryopreservation."

"Penelitian dilakukan untuk mengetahui pengaruh kombinasi gliserol 6% dengan beberapa konsentrasi kuning telur terhadap kualitas spermatozoa ikan kerapu kertang Epinephelus lanceolatus (Bloch, 1970) pascakriopreservasi. Semen ikan kerapu kertang didapatkan dengan metode pengurutan. Larutan pengencer terdiri atas marine fish ringer, gliserol 6%, dan berbagai konsentrasi kuning telur (0%; 5%; 10%; 15%; 20%; dan 25%). Ekuilibrasi dilakukan selama 10 menit pada suhu 4 ºC. Pembekuan dalam freezer (-20 ºC) selama 48 jam. Pencairan pada suhu 45 ºC selama 30-60 detik. Evaluasi semen dilakukan secara makroskopis (warna, volume, dan pH) dan secara mikroskopis (motilitas, viabilitas, dan abnormalitas) serta kemampuan fertilisasinya terhadap telur ikan kerapu macan. Berdasarkan hasil uji ANAVA yang dilanjutkan dengan uji Tukey, terdapat perbedaan nyata (P>0,05) terhadap motilitas, viabilitas, dan kemampuan fertilisasi spermatozoa ikan kerapu kertang pascakriopreservasi, akan tetapi tidak berpengaruh nyata terhadap abnormalitas spermatozoa pascakriopreservasi (P>0,05). Hasil penelitian menunjukkan bahwa perlakuan kuning telur 15% merupakan konsentrasi optimum karena menghasilkan nilai persentase rata-rata motilitas, viabilitas, dan kemampuan fertilisasi tertinggi masing-masing sebesar 83,64 ± 1,72%, 81,44 ± 2,06%, dan 77,31 ± 1,90%. Nilai rata-rata terendah pada persentase abnormalitas sebesar 21,50 ± 1,20%.

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Research on the effect of combination 6% glycerol and eggyolk as cryoprotectant of giant grouper Epinephelus lanceolatus (Bloch, 1970) spermatozoa quality postcryopreservation. Giant grouper cement is obtained by sorting method. The diluent solution used consist of marine fish ringer, 6% glycerol, and varian egg yolk concentrations (0%; 5%; 10%; 15%; 20%; dan 25%). Equilibration is carried out for 10 minutes at 4ºC. Freezing in the freezer (-20 ºC) for 48 hours. Thawing at 45 ºC for 30-60 seconds. Cement evaluation is carried out macroscopically (color, volume, and pH) and microscopically (motility, viability, and abnormality) and also the ability to fertilize the egg of tigger grouper. Based on the ANAVA statistical test followed by Tukey, there were significant differences effect on motility, viability, and fertilization ability of spermatozoa of giant grouper postcryopreservation (P> 0.05), but abnormality was significantly affected (P <0.05). (P> 0.05). The optimum concentration of egg yolk is 15% because it produces the highest percentage value of motility, viability, and fertilization ability of 83.64 ± 1.72%, 81.44 ± 2.06% and 77.31 ± 1.90% respectively. The lowest average value of abnormalitiy percentage is 21.50 ± 1.20%."

"Penelitian kriopreservasi spermatozoa ikan kerapu kertang memiliki tujuan mengetahui pengaruh berbagai konsentrasi susu skim (0%, 5%, 10%, 15%, 20%, 25%) yang dikombinasikan dengan gliserol 6% terhadap motilitas, viabilitas, dan abnormalitas serta kemampuan fertilisasi spermatozoa ikan kerapu kertang pascakriopreservasi terhadap sel telur ikan kerapu macan. Larutan pengencer yang digunakan dalam penelitian adalah larutan marine fish Ringer, gliserol 6%, susu skim berbagai konsentrasi. Rasio pengenceran yang digunakan adalah 1:9. Kriopreservasi dilakukan dalam freezer pada suhu -20°C, dengan lama penyimpanan selama 48 jam. Spermatozoa hasil kriopreservasi selama 48 jam digunakan untuk membuahi sel telur ikan kerapu macan. Hasil fertilisasi digunakan untuk mengukur parameter kualitas spermatozoa yang baik. Hasil uji ANAVA satu arah menunjukkan pemberian berbagai konsentrasi susu skim memiliki nilai rata-rata persentase motilitas, viabilitas, dan abnormalitas spermatozoa ikan kerapu kertang 48 jam pascakriopreservasi yang berbeda nyata (P˃0,05). Hasil terbaik ditunjukkan pada konsentrasi susu skim 20% dengan nilai persentase motilitas, viabilitas, dan abnormalitas secara berurutan sebesar 80,51 ± 3,46%; 81,24 ± 2,34%; dan 25,35 ± 2,04 %. Hasil analisis pada fertilisasi spermatozoa pascakriopreservasi menyatakan bahwa nilai rata-rata persentase fertilisasi tidak berbeda nyata antar perlakuan, namun pada konsentrasi susu skim 20% memberikan kemampuan fertilisasi yang baik yaitu 68 ± 1,70%.

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The objective of this study was to discover the effect of various concentration skim milk from (0%, 5%, 10%, 15%, 20%, and 25%) and combined with glycerol 6% which give the best effect towards motility, viability, abnormality and fertilization for Ephinephelus fuscogutattus (Forsskal 1775) capability of Ephinephelus lanceolatus (Bloch 1970) spermatozoa 48 hour after freezing. We used marine fish Ringer, glycerol 6%, and various concentration skim milk. Dilute the spermatozoa at 1 : 9 ratio. Cryopreservation is carried out in a freezer with a temperature of -20°C with a storage time of 48 hours. Cryopreservation spermatozoa for 48 hours is used to fertilize the egg Ephinephelus fuscogutattus (Forsskal 1775). Fertilization results are used to measure the quality parameter of spermatozoa Ephinephelus lanceolatus (Bloch 1970). Based on the ANAVA analysis, the treatment groups showed significant difference in average motility, viability and spermatozoa abnormality percentage with the control (P˃0.05). ANAVA analysis showed best result are obtained from 20% skim milk with average motility (80.51 ± 3.46%), sperm viability (81.24 ± 2.34%), and sperm abnormality (25.35 ± 2.04%). The results of analysis on fertilization with sperm post- cryopreservation stated that the average value of the percentage of fertilization was not significantly different between treatments, but at the concentration of skim milk 20% give a best ability of fertilization which was 68 ± 1.70%."

"Penelitian pengaruh pemberian berbagai konsentrasi kuning telur ayam kampung terhadap kualitas spermatozoa ikan koi Cyprinus carpio, Linnaeus 1758 48 jam pascakriopreservasi dilakukan untuk mengetahui konsentrasi optimum kuning telur ayam kampung yang dapat digunakan untuk menjaga kualitas spermatozoa ikan koi. Semen ikan koi diperoleh dengan cara stripping pada bagian urogenital ikan koi. Evaluasi semen ikan koi dilakukan secara makroskopis volume, pH, dan warna semen dan mikroskopis persentase motilitas, viabilitas, dan abnormalitas. Penelitian menggunakan larutan pengencer metanol 10 dan berbagai konsentrasi kuning telur ayam kampung dengan perbandingan semen dan pengencer 1:4. Perlakuan yang diberikan yaitu kuning telur ayam kampung dengan konsentrasi 0 kontrol, 5, 10, 15, 20, dan 25. Pembekuan dilakukan pada suhu -34 oC selama 48 jam. Data yang diperoleh menunjukkan distribusi normal dan bervariasi homogen setelah dilakukan uji normalitas Shapiro-Wilk dan uji homogenitas Levene. Hasil uji Analisis Variansi ANAVA satu arah menunjukkan pengaruh perlakuan kuning telur ayam kampung.

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Research about various effect of free range chicken egg yolk concentration to spermatozoa quality of Koi Fish Cyprinus carpio, Linnaeus 1758 48 hours post cryopreservation was done to know optimum concentration of free range chicken egg yolk that can still keep the quality of koi fish spermatozoa. Milt sperm koi fish was collected by stripping at urogenital area of the fish. Koi fish sperm evaluation was done macroscopically volume, pH, milt color and microscopically percentage of motility, viability, and abnormality of spermatozoa.

The research was using dilution solution methanol 10 and various concentrations of free range chicken egg yolk with ratio 1 4. The given concentration of free range chicken egg yolk was 0 control, 5, 10, 15, 20, and 25. Freezing was done at 34 oC for 48 hours. The data obtained showed normal and homogenous after processed with the Shapiro Wilk normality test and Levene homogenity test. The one factor ANOVA showed that various concentration of free range chicken egg yolk had effect."

"Telah dilakukan penelitian mengenai kriopreservasi ikan koi Cyprinus carpio, Linnaeus 1758 menggunakan berbagai konsentrasi susu skim yang dikombinasikan dengan metanol 10. Tujuan penelitian adalah mengetahui pengaruh konsentrasi susu skim 0, 5, 10, 15, 20, 25 yang dikombinasikan dengan metanol 10 terhadap kualitas spermatozoa ikan koi 24 jam pascakriopreservasi. Hasil uji ANAVA satu faktor menunjukkan pemberian berbagai konsentrasi susu skim memiliki nilai rata-rata persentase motilitas, viabilitas, dan abnormalitas spermatozoa ikan koi 24 jam pascakriopreservasi yang berbeda nyata.

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Cryopreservation of koi Cyprinus carpio, Linnaerus 1758 spermatozoa using various concentrations of skim milk combined with 10 methanol has been studied. The objective of the study was to know the effect of skim milk in various concentration on spermatozoa quality of koi Cyprinus carpio, Linnaeus 1758 postcryopreservation. The one factor ANOVA showed that various concentrations of skim milk had a significant difference."

"Telah dilakukan penelitian mengenai kriopreservasi ikan koi Cyprinus carpio, Linnaeus 1758 dengan menggunakan berbagai konsentrasi madu yang dikombinasikan dengan metanol 10 . Tujuan penelitian yaitu untuk mengetahui pengaruh konsentrasi madu 0 , 0,1 , 0,3 , 0,5 , 0,7 , 0,9 terhadap kualitas spermatozoa ikan koi 24 jam pascakriopreservasi. Hasil uji ANAVA satu faktor menunjukkan pemberian berbagai konsentrasi madu memiliki nilai rata-rata persentase motilitas, viabilitas, dan abnormalitas spermatozoa ikan koi 24 jam pascakriopreservasi yang berbeda nyata.

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Cryopreservation of koi Cyprinus carpio, Linnaerus 1758 spermatozoa using various concentrations of honey combined with 10 methanol has been studied. The objective of the study was to know the effect of honey in various concentration 0 , 0,1 , 0,3 , 0,5 , 0,7 , 0,9 on spermatozoa quality of koi Cyprinus carpio, Linnaeus 1758 24 hours postcryopreservation. The one factor ANOVA showed that various concentrations of honey had a significant difference."

"ABSTRAKKriopreservasi merupakan metode penyimpanan materi genetik pada suhu yang rendah dalam jangka waktu yang lama. Manfaat kriopreservasi salah satunya untuk mengatasi masalah penurunan populasi. Ikan kancra merupakan salah satu spesies dari famili Cyprinidae yang populasinya semakin menurun sehigga diperlukan upaya pelestarian. Faktor yang mempengaruhi keberhasilan kriopreservasi salah satunya adalah krioprotektan. Krioprotektan alami digunakan karena memiliki toksisitas yang lebih rendah dibanding krioprotektan dari bahan kimia. Kuning telur merupakan krioprotektan alami yang dapat menjaga sel selama proses kriopreservasi. Tujuan dari penelitian yaitu mengevaluasi efek kuning telur ayam kampung sebagai krioprotektan alami terhadap motilitas, viabilitas, abnormalitas dan fertilisasi spermatozoa 48 jam pascakriopreservasi. Konsentrasi kuning telur ayam kampung yang digunakan yaitu: 0%, 5%, 10%, 15%, 20% dan 25%. Rasio yang digunakan untuk pengenceran sperma yaitu 1:10. Analisis data menggunakan uji ANAVA kemudian dilanjutkan dengan uji Tukey. Berdasarkan hasil uji ANAVA satu arah diketahui bahwa krioprotektan kuning telur ayam kampung yang dikombinasikan dengan metanol 10% memberikan pengaruh (P<0,05) terhadap motilitas, viabilitas, abnormalitas dan fertilitas spermatozoa pascakriopreservasi. Konsentrasi kuning telur 5% merupakan konsentrasi optimum yang dapat mempertahankan persentase motilitas, viabilitas dan fertilitas tertinggi masing-masing 84,06 ± 1,67%, 82,13 ± 1,75% dan  92,96 ± 1,94% serta nilai persentase abnormalitas terendah 25,25 ± 2,22%.

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ABSTRACTCryopreservation is a method of storing genetic material at low temperatures for long periods of time. One of the benefits of cryopreservation is to overcome the problem of population decline. The kancra fish is one of the species of the Cyprinidae family whose population is declining so that conservation efforts are needed. One of the factors that influence the success of cryopreservation is cryoprotectant. Natural cryoprotectants are used because they have lower toxicity than cryoprotectants from chemicals. Egg yolk is a natural cryoprotectant that can maintain cells during the cryopreservation process. The purpose of this study is to evaluate the effect of egg yolk as natural cryoprotectants on motility, viability, abnormalities, and fertilization of 48-hour spermatozoa after cryopreservation. Concentrations of free-range chicken egg yolk used are: 0%, 5%, 10%, 15%, 20% and 25%. The ratio used for sperm dilution is 1:10. Data analysis using the ANAVA test and then followed by the Tukey test. Based on the results of the one-way ANAVA test, it was found that cryoprotectant of free-range chicken egg yolk combined with 10% methanol had an effect (P <0.05) on motility, viability, abnormality, and fertility of post-cryoprotected spermatozoa. Egg yolk concentration of 5% was the optimum concentration that can maintain the highest percentage of motility, viability and fertility, respectively 84.06 ± 1.67%, 82.13 ± 1.75% and 92.96 ± 1.94% and the percentage value the lowest abnormality was 25.25 ± 2.22%."