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KRIOPRESERVASI SEMEN

KRIOPRESERVASI SEMEN

Kriopreservasi semen merupakan proses penurunan suhu, dehidrasi sel, pembekuan, penyimpanan dan kemudian pencairan kembali. Kriopreserasi semen memegang peranan penting dalam pelaksanaan inseminasi buatan karena dapat mengakselerasi penyebaran keragaman genetik dan memfasilitasi distribusi superior genetik keseluruh wilayah. Proses kriopreservasi semen yang baik akan meningkatkan efisiensi dan tingkat kesuksesan dalam program inseminasi buatan (Ugur et al., 2019) Program inseminasi buatan sepenuhnya bisa dilaksanakan hanya dengan semen beku yang memungkinkan penyimpanan yang lebih lama dan cakupan distribusi yang tidak terbatas (Layek et al., 2016). Manfaat kriopreservasi semen diantaraya: efisiensi penggunaan pejantan, inseminasi buatan tidak dibatasi waktu dan tempat, memudahkan transportasi, lebih ekonomis, dan mempunyai daya simpan yang panjang.

Semen beku merupakan semen yang telah mengalami proses pengenceran dengan bahan pengencer kemudian ditambahkan glicerol dalam proses pembekuannya. Pembekuan semen dapat dilakukan secara bertahap atau langsung (vitrifikasi), pembekuan semen dengan menggunakan medium nitrogen cair (N2 cair) dengan suhu -196oC. Spermatozoa yang telah dibekukan akan mampu bertahan hidup dalam kurun waktu yang lama bahkan setelah 30 tahun baru akan mengalami penurunan kualitasnya (Ismaya, 2014). Proses kriopreservasi tersebut berakibat pada kerusakan pada spermatozoa sehingga berdampak pada fertilitas. Layek et al. (2016) mengungkapkan bahwa kerusakan akibat kriopreservasi bisa mencapai 50%. Kerusakan spermatozoa dapat berupa kerusakan membran plasma, membran akrosom, dan kerusakan biokimia dalam sel seperti penurunan aktivitas proteolik akrosom (Ismaya, 2014). Beberapa penyebab kerusakan spermatozoa saat proses pembekuan diantaranya cekaman dingin, dan terbentuknya kristal es (Watson, 1995; Watson, 2000; Susilawati, 2011).

Cekaman Dingin

Cekaman dingin atau Cold shock merupakan penyebab utama penurunan kualitas semen (Ugur, 2019). Spermatozoa kehilangan kekuatan membran dan fungsinya ketika terjadi penurunan suhu 0oC -20oC. Fenomena tersebut terjadi karena terjadi perubahan fase membran lipid sehingga membran lipid kehilangan kemampuan permeabilitas. Cekaman dingin diketahui bedampak pada penurunan kemampuan kinetik ekor spermatozoa sehingga terjadi penurunan motilitas (Watson, 1995).

Penambahan pengencer yang berisi krioprotektan sebelum proses pembekuan dapat mengurangi resiko penurunan kualitas semen akibat dari cold shock. Saat ini, pengencer yang berbasis kuning telur paling umum digunakan dalam proses kriopreservasi berbagai macam ternak. Kandungan Low Density Lipoprotein (LDL) dapat melindungi spermatozoa dari penurunan kualitas semen seperti motilitas, kekuatan membran, dan kemampuan fertilisasi (Bencharif et al., 2010).

Pembentukan Krista Es dan Cekaman Osmotik

Penurunan suhu melewati 0oC larutan ekstraseluler dan intraseluler membentuk kristal es, sehingga dapat menyebabkan kerusakan seara mekanis terhadap membran plasma (Watson, 1995). Waktu pembekuan yang tepat memegang peranan penting dalam meminimalisir kerusakan sel spermatozoa. Pembekuan lambat meminimalisir kerusakan mekanis yang disebabkan oleh kristal es. Namun, kerusakan terjadi akibat efek padatan yang diakibatkan terjadinya perbedaan tekanan osmotik sehingga fase cairan dalam sel keluar menyisakan fase padatan dalam sel. Sebaliknya, pembekuan cepat mencegah kerusakan sel spermatozoa akibat dari efek padatan namun tetap terjadi kerusakan mekanis akibat dari pembentukan kristal es berukuran besar  (Hafez, 2000).

Penambahan krioprotektan (gliserol) dapat menurunkan resiko kerusakan spermatozoa. Penambahan krioprotektan akan menurunkan titik beku sel spermatozoa hingga -196oC. Mekanisme perubahan titik beku disebabkan masuknya krioprotektan ke dalam sel karena sifatnya yang higroskopis, kemudian menggantikan cairan dalam sel dengan cairan krioprotektan tersebut sehingga dapat meminimalisir kerusakan  (Susilawati, 2011).

Ketahanan spermatozoa terhadap proses pembekuan dapat dipengaruhi oleh berbagai faktor seperti efek dari temperatur dan pengaruh musim (Kiovisto, 2016). Badan Standarisasi Nasional (2017) mensyaratkan bahwa semen beku yang layak digunakan memiliki motilitas lebih dari atau sama dengan 40%.

 

DAFTAR PUSTAKA

Bencharif, D., L. Amirat-Briand, A. Garand, M. Anton, E. Schmitt, S. Desherces, G. Delhomme, M.L. Langlois, P. Barrière dan S. Destrumelle, 2010. Freezing Canine Sperm: Comparison of Semen Extenders Containing

Hafez E.S.E. 2000. Preservation and Cryopreservation of Gametes and Embryos. In. Hafez E.S.E (ed). Reproduction in Farm Animals 7th Ed. Lippincott Williams & Wilkins, USA.

Ismaya. 2014. Bioteknologi Inseminasi Buatan pada Sapid dan Kerbau. Gadjah Mada University Press, Yogyakarta.

Kiovisto, MB, MTA Costa, SHV Perri and WRR Vicente. 2008.The Effect of Season on Semen Characteristics and Freezability in Bos indicus and Bos taurus Bulls in the Southeastern Region of Brazil. Reprod. Dom. Anim. 44: 587–592 .

Layek, S.S., T.K. Mohanty , A. Kumaresan , J.E. Parks  . 2016. Cryopreservation of Bull Semen: Evolution from Egg Yolk Based to Soybean Based Extenders. Anim. Reprod. Sci.  172 : 1–9.

Susilawati, T.. 2011. Spermatology. UB Press, Malang.

Ugur M. R., A. S. Abdelrahman, H C. Evans, A. A. Gilmore, M. Hitit, L. A. Arifiantini, B. Purwantara, A. Kaya, dan E. Memili. 2019. Advances In Cryopreservation Of Bull Sperm. Front. Vet. Sci. 6:268.

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