spermatozoa, aktivitas ovum, pembentukan pronukleus jantan dan betina serta pertautan kromosommaternal paternal membentuk genom (Elder dan Dale, 2011). Tingkat fertilisasi adalah jumlah oosit yang mempunyai 2 pronukleus (2PN) yaitu jantan dan betina.
Tabel 3. Perhitungan tingkat tahapan maturasi dan fertilisasi oosit pada berbagai perlakuan Perlakuan Jumlah oosit Tingkat Pematangan Inti Pembentukan Pronukleus (%) Total M-II Tingkat Fertilisasi (M-IIxTingkat Fertilisasi: 100%) P0 36 (72.2) (22.2) (16.02) P1 26 (80.8) (42.3) (34.17) P2 34 (61.7) (38.2) (23.56 P3 32 (59.4) (43.8) (26.01)
Keterangan : P0 (Kontrol, tanpa ITS), P1 (penambahan ITS 5 ng/ml pada media maturasi), P2 (penambahan ITS 10 ng/ml pada media maturasi), P3 (penambahan ITS 15 ng/ml pada media). M-II: metaphase II, tingkat fertilisasi : gabungan 2 pn dan >2PN.
Berdasarkan persentase tingkat maturasi dan fertilisasi oosit bahwa perlakuan
yang terbaik adalah perlakuan yang menghasilkan nilai tinggi dari perkalian tingkat
maturasi dan tingkat fertilisasi. Dalam hal ini adalah P1 yang menghasilkan nilai
29 KESIMPULAN DAN SARAN
Kesimpulan
Penambahan Insulin Transferrin Selenium (ITS) pada medium sebanyak 5 – 15 ng/ml tidak berpengaruh nyata terhadap tingkat maturasi dan fertilisasi oosit sapi Bali
secara in vitro.
Saran
Sebaiknya ada kajian ilmiah lebih lanjut untuk mengetahui pengaruh penambahan Insulin Transferrin Selenium (ITS) pada medium terhadap tingkat maturasi dan fertilisasi oosit sapi Bali secara in vitro.
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36 LAMPIRAN
Lampiran 1. Komposisi Media Maturasi Oosit Secara In Vitro
No Nama Bahan Volume
1 TCM-199 1800 µl
2 Serum FBS (fetal bovine serum) 200µl
3 PMSG (pregnant mare serum gonadotropin) 20µl
4 hCG(human chorionic gonadotropin) 20µl
5 Gentamycin 4µl
Lampiran 2. Komposisi Media Fertilisasi Secara In Vitro
No Nama Bahan Volume
1 Ultra pure water 50 ml
2 NaCl (natrium/sodium chloride) 0,2629 gram
3 KCL (kalium chloride) 0,0447 gram
4 NaHCo3 (natrium bicarbonate) 0,1050 gram
5 NaH2PO4 (natrium dihydrogen phosphate monohydrate) 0,0030 gram 6 MgSO47H2O (magnesium sulfat-heptahydrate) 0,0061 gram
7 Sodium lactate 60% syrup 0,095 ml
8 Hepes 0,1191 gram
9 CaCl22H2O (calcium chloride_dihydrate) 0,0588 gram
10 Sodium pyruvate 0,0110 gram
11 Caffeine anhydrous 0,0194 gram
12 BSA (fatty acid free) fraksi V 0,2500 gram
13 Gentamycin 10 µl
Lampiran 3. Jumlah oosit dari beberapa kualitas yang berbeda
Ulangan Perlakuan (Kualitas Oosit) ITS Jumlah Kontrol 5ng/ml 10ng/ml 15ng/ml 1 10 10 10 10 40 2 10 10 10 10 40 3 10 10 10 10 40 4 10 10 10 10 40 Total 40 40 40 40 180
37 Lampiran 4. Penggunaan ITS
5 mg/ml (ITS)
250 µL(ITS)
250 µL ITS + 1000 µL TCM
38 LAMPIRAN 5. DATA TINGKAT MATURASI PENAMBAHAN ITS DENGAN KONSENTRASI BERBEDA (%)
Perlakuan Ulangan Jumlah Oosit Lose Oosit GV GVBD M I M II
A (Kontrol) 1 10 7 0 (0.0) 0 (0.0) 1 (0.0) 2(20.0) 2 10 6 0 (0.0) 0 (0.0) 2 (20.0) 2 (20.0) 3 10 5 0 (0.0) 1 (10.0) 3 (30.0) 2 (20.0) 4 10 5 0 (0.0) 1 (10.0) 2 (20.0) 2 (20.0) Total 40 23 0 (00.0) 2 (20.0) 8 (80.0) 8 (80.0) B (5 ng/ml) 1 10 7 0 (0.0) 0 (0.0) 1 (10.0) 2 (20.0) 2 10 7 0 (0.0) 0 (0.0) 2 (20.0) 1 (10.0) 3 10 6 0 (0.0) 0 (0.0) 0 (00.0) 4 (40.0) 4 10 7 0 (0.0) 0 (0.0) 2 (20.0) 1 (10.0) Total 40 27 0 (0.0) 0 (00.0) 5 (50.0) 8 (80.0) C (10 ng/ml) 1 10 6 1 (10.0) 0 (0.0) 2 (20.0) 1 (10.0) 2 10 6 1 (10.0) 1 (10.0) 1 (10.0) 1 (10.0) 3 10 5 1 (10.0) 0 (0.0) 3 (30.0) 1 (10.0) 4 10 6 1 (10.0) 2 (20.0) 0 (0.0) 1 (10.0) Total 40 23 4 (40.0) 3 (30.0) 8 (80.0) 4 (40.0) D (15 ng/ml) 1 10 7 0 (0.0) 1 (10.0) 1 (10.0) 1 (10.0) 2 10 5 0 (0.0) 2 (20.0) 2 (20.0) 1 (10.0) 3 10 6 1 (10.0) 1 (10.0) 2(20.0) 0 (0.0) 4 10 6 1 (10.0) 1 (10.0) 1 (10.0) 1 (10.0) Total 40 24 2 (2.0) 5 (50.0) 6 (60.0) 3 (30.0)
39 LAMPIRAN 6. DATA TINGKAT FERTILISASI PENAMBAHAN ITS DENGAN KONSENTRASI BERBEDA (%)
Perlakuan Ulangan Jumlah Oosit Lose Oosit PN-0 PN-1 PN-2 >PN-2
A (Kontrol) 1 10 7 0 (0.0) 1 (10.0) 0 2(20.0) 2 10 6 2 (20.0) 0 (0.0) 1 (10.0) 1 (10.0) 3 10 5 4 (40.0) 0 (0.0) 0 (0.0) 2 (20.0) 4 10 5 2 (20.0) 1 (10.0) 2 (20.0) 0 (0.0) Total 40 23 8 (80.0) 2 (20.0) 3 (30.0) 5 (50.0) B (5 ng/ml) 1 10 7 0 (0.0) 0 (0.0) 1 (10.0) 2 (20.0) 2 10 7 0 (0.0) 1 (10.0) 2 (20.0) 0 (0.0) 3 10 6 0 (0.0) 0 (0.0) 2 (20.0) 2 (20.0) 4 10 7 0 (0.0) 1 (10.0) 1 (10.0) 1 (10.0) Total 40 27 0 (0.0) 2 (20.0) 6 (60.0) 5 (50.0) C (10 ng/ml) 1 10 6 0 (0.0) 1 (10.0) 2 (20.0) 1 (10.0) 2 10 6 1 (10.0) 0 (0.0) 1 (10.0) 2 (20.0) 3 10 5 0 (0.0) 0 (0.0) 3 (30.0) 2 (20.0) 4 10 6 1 (10.0) 1 (10.0) 1 (10.0) 1 (10.0) Total 40 23 2 (20.0) 2 (20.0) 7 (70.0) 6 (60.0) D (15 ng/ml) 1 10 7 0 (0.0) 0 (0.0) 1 (10.0) 2 (20.0) 2 10 5 0 (0.0) 1 (10.0) 1 (10.0) 3 (30.0) 3 10 6 0 (0.0) 0 (0.0) 1(10.0) 3 (30.0) 4 10 6 0 (0.0) 1 (10.0) 2 (20.0) 1 (10.0) Total 40 24 0 (0.0) 2 (20.0) 5 (50.0) 9 (90)
47 A=B
A=C
A=D LAMPIRAN 7. ANALISIS 2X2 CONTINGENCY TABLE DATA TINGKAT MATURASI
47 B=A
B=C
47 C=A
C=B
47 D=A
D=B
47 A=B
A=C
A=D LAMPIRAN 8. ANALISIS 2X2 CONTINGENCY TABLE DATA TINGKAT FERTILISASI
47 B=A
B=C
47 C=A
C=B
47 D=A
D=B
48 Lampiran 9. Dokumentasi Penelitian
Ovarium sapi bali Oosit setelah di fertilisasi
Oosit setelah direkatkan dengan vaselin Preparat oosit kualitas A, B dan C
Preparat yang difiksasi Proses pencacahan
49
Bahan untuk fiksasi Bahan untuk pewarnaan
Tissu dan alkohol 70% untuk sterilisasi Disk maturasi dan fertilisasi
50
Scalpel, pinset dan gunting bedah Minitube
Oven untuk sterilisasi kering Inkubator
Timbangan analitik Stirrer
51 Pipet yang dimodifikasi Bunsen untuk memodifikasi pipet
Mikro pipet dan tip Pipet volumentrik
52 RIWAYAT HIDUP
Hikmayani Iskandar lahir pada tanggal 25 Juni 1996 di Makassar, Provinsi Sulawesi Selatan. Penulis merupakan anak dari pasangan Bapak Iskandar dan Ibu Ramliah. Pendidikan formal yang telah ditempuh oleh penulis yakni: SD Muhammadiyah II Makassar 2001 - 2007; SMP Muhammadiyah 09 Makassar Tahun 2007 - 2010 ; SMA Negeri 22 Makassar Tahun 2010 - 2013 dan pada tahun 2013 - 2017 penulis melanjutkan pendidikannya di Fakultas Peternakan Program Studi Ilmu Peternakan Universitas Hasanuddin (Unhas) Makassar. Selama menjadi mahasiswa penulis aktif sebagai pengurus organisasi Himpunan Mahasiswa Produksi Ternak Universitas Hasanuddin (HIMAPROTEK-UNHAS), BEM Peternakan, dan UKM Tenis Meja. Penulis juga aktif sebagai asisten pada mata kuliah Fisiologis Ternak (2015-2017), Ternak Unggas (2015-2017), dan Manajemen Ternak Unggas (2015-2016).