Rophi, Apriani Herni (2018) Pengaruh Penambahan Ekstrak Buah Mengkudu (Morinda Citrifolia L. ) Dalam Pengencer Tris Kuning Telur Terhadap Kualitas Spermatozoa Kambing Setelah Pembekuan Pada -80 °C. Magister thesis, Universitas Brawijaya.
Abstract
Dalam proses kriopreservasi proses pembekuan dan pencairan kembali (post thawing) dapat menyebabkan kejutan dingin (cold shock) yang memicu produksi Reactive Oxygen Species (ROS). Produksi ROS yang berlebihan akan menyebabkan terjadinya peroksidasi lipid pada membran sel. Peroksidasi lipid akan merusak struktur matriks lipid membran dan menurunkan fluiditas membran sel, yang menyebabkan fungsi membran terganggu sehingga berakibat pada penurunan motilitas, viabilitas, serta peningkatan abnormalitas sel spermatozoa. Untuk mencegah terbentuknya ROS secara berlebihan maka dalam pengenceran spermatozoa perlu ditambahakan antioksidan. Mengkudu (Morinda citrifolia L.) merupakan tanaman yang dapan berfungsi sebagai sumber antioksidan karena mengandung vitamin C, flavonoid dan fenol. Diharapkan dengan pemberian ekstrak buah mengkudu dalam proses kriopreservasi dapat mempertahankan keutuhan membran plasma sehingga motilitas, viabilitas spermatozoa tetap terjaga serta abnormalitas sel dapat dikurangi. Penelitian ini bertujuan untuk mengetahui pengaruh media mengencer tris kuning telur yang ditambah ekstrak buah mengkudu (M. citrifolia L.) terhadap kualitas spermatozoa (motilitas, viabilitas abnormalitas, serta ultrastruktur membran) setelah dibekukan pada suhu -80°C selama 24 jam. Penelitian menggunakan metode ekperimental dengan Rancangan Acak Lengkap (RAL). Penelitian dibuat dalam 4 perlakuan dan masing-masing perlakukan dilakukan sebanyak 6 kali ulangan, yaitu: P0 (sebagai kontrol tris, kuning telur, gliserol 7%); P1 (tris, kuning telur, gliserol 7% dan ekstrak mengkudu 10%); P2 (tris, kuning telur, gliserol 7% dan ekstrak mengkudu 20%); serta P3 (tris, kuning telur, gliserol 7% dan ekstrak mengkudu 30%). Semen hasil penampungan diencerkan menggunakan tris kuning telur yang ditambahkan ekstrak buah mengkudu sesuai dengan perlakuan kemudian diekuilibrasi dalam cooltop pada suhu 5°C selama 2 jam yang disertai dengan penambahan gliserol sebanyak 2 kali yaitu setelah satu jam ekuilibrasi dan satu jam berikutnya setelah penambahan yang pertama. Semen cair hasil ekuilibrasi disimpan menggunakan Mr FrostyTM kemudian dibekukan dalam ultrafreezer bersuhu -80 °C, selama 24 jam. Semen di thawing pada waterbath bersuhu 37 °C selama 2,5 menit. Evaluasi kualitas spermatozoa dilakukan pada semen segar, semen cair, dan semen post thawing. Data yang diperoleh diolah menggunakan program SPSS (versi 16.0) dengan analysis of variant (ANOVA) dan Uji Duncan’s digunakan untuk mengetahui perbedaan nilai yang diperoleh, perbedaan dianggap signifikan pada p < 0.05. Hasil penelitian menunjukkan bahwa persentase motilitas spermatozoa post thawing pada kelompok kontrol (P0), P1, P2 dan P3 secara berturut-turut adalah 2,17 %, 6,33 %, 10,83% dan 9,67 %. Persentase viabilitas spermatozoa pada kelompok kontrol (P0), P1, P2 dan P3 secara berturut-turut adalah 9,00 %, 11,33 %, 22,00 % dan 22,17 %. Pesentase abnormalitas pada kelompok kontrol (P0), P1, P2 dan P3 secara berturut-turut adalah 8,27%, 6,83 %, 5,77 % dan 10,67 %. Jenis abnormalitas spermatozoa yang ditemukan pada semen segar meliputi broke tail, “dag”defect, proximal droplet, coiled tail, shoehook tail, bent tail, dan distal droplet. Jenis abnormalitas yang ditemukan pada semen post thawing meliputi broke tail, absent head, deteched head, “dag”defect, tapered head, proximal droplet, round head, abaxial, coiled tail, shoehook tail, bent tail, dan distal droplet. Hasil pengamatan ultrastruktur spermatozoa post thawing menunjukkan adanya kerusakan pada membran kepala spermatozoa bagian anterior, yang tampak tidak rata dan seperti berlubang. Dari hasil penelitian dapat disimpulkan bahwa penambahan dosis 20% (P2) ekstrak buah mengkudu dalam pengencer tris kuning telur merupakan dosis paling baik untuk mempertahankan kualitas spermatozoa setelah pembekuan pada -80 °C.
English Abstract
In the cryopreservation process the freezing process and thawing can cause cold shock which triggers the production of Reactive Oxygen Species (ROS). Excessive ROS production will cause lipid peroxidation in the cell membrane. Lipid peroxidation will damage the structure of the membrane lipid matrix and reduce the fluidity of cell membranes, which causes disturbed membrane function resulting in decreased motility, viability, and increased sperm cell abnormalities. To prevent excessive formation of ROS, antioxidants need to be added to the spermatozoa. Noni (Morinda citrifolia L.) is a plant that can function as a source of antioxidants because it contains vitamin C, flavonoids and phenols. It is expected that by giving out noni fruit in the cryopreservation process can maintain the integrity of the plasma membrane so that motility, viability of spermatozoa is maintained and cell abnormalities can be reduced. The aim of this study was to determine the effect of media on tris egg yolk extender added with noni fruit (M. citrifolia L.) extract on sperm quality (motility, abnormal viability, and membrane ultrastructure) after being frozen at -80 ° C for 24 hours. The study used experimental methods with a completely randomized design (CRD). The study was made in 4 treatments and 6 repetitions, namely: P0 (as a control consisting of: tris, egg yolk, glycerol 7%); P1 (consisting of: tris, egg yolk, glycerol 7% and noni extract 10%); P2 (consisting of: tris, egg yolk, glycerol 7% and noni extract 20%); and P3 (consisting of: tris, egg yolks, 7% glycerol and 30% noni extract). Semen from the reservoir was diluted using egg yolk tris added with noni fruit extract according to the treatment then equilibrated in cooltop at 5 ° C for 2 hours accompanied with the addition of glycerol 2 times after one hour of equilibration and one hour after the first addition, the equilibrium liquid semen was stored using Mr. FrostyTM then frozen in utrafreezer at -80 ° C, for 24 hours. The semen is thawed on the waterbath at 37 ° C for 2.5 minutes for 2.5 minutes. Evaluation of the quality of spermatozoa was carried out on fresh semen, liquid semen and post thawing semen. The data obtained were processed using the SPSS program (version 16.0) with analysis of variant (ANOVA) and Duncan's test was used to determine the difference in value obtained, the difference was considered significant at p <0.05. The results showed that the percentage of sperm motility post-thawing in the control group (P0), P1, P2 and P3 were 2.17%, 6.33%, 10.83% and 9.67% respectively. Spermatozoa viability percentage in the control group (P0), P1, P2 and P3 were 9.00%, 11.33%, 22.00% and 22.17% respectively. The percentage of abnormalities in the control group (P0), P1, P2 and P3 were 8.27%, 6.83%, 5.77% and 10.67% respectively. Types of sperm abnormalities found in fresh semen include the broke tail, "dag" defect, proximal droplet, coiled tail, shoehook tail, bent tail, and distal droplet. The types of abnormalities found in post thawing semen include the broke tail, absent head, deteched head, "dag" defect, tapered head, proximal droplet, round head, abaxial, coiled tail, shoehook tail, bent tail, and distal droplet. SEM observation found that there was spermatozoa membrane damage marked a rought surface and like a hole in the membrane of anterior sperm cell. Based on the results of the study it can be concluded that the 20% dose (P2) of noni fruit extract in tris egg yolk extender is the optimum dose to maintain the quality of spermatozoa after freezing at -80 °C.
Other obstract
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| Item Type: | Thesis (Magister) |
|---|---|
| Identification Number: | TES/581.7/ROP/2018/p/041809343 |
| Uncontrolled Keywords: | PLANT ECOLOGY |
| Subjects: | 500 Natural sciences and mathematics > 581 Specific topics in natural history of plants > 581.7 Plant ecology, plants characteristic of specific of environments |
| Divisions: | S2/S3 > Doktor Biologi, Fakultas MIPA |
| Depositing User: | Budi Wahyono Wahyono |
| Date Deposited: | 03 Sep 2019 07:01 |
| Last Modified: | 03 Sep 2019 07:01 |
| URI: | http://repository.ub.ac.id/id/eprint/172329 |
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