Bibos banteng) merupakan plasma

BAB VI. RINGKASAN

Sapi Bali (Bos sondaicus, Bos javanicus, Bos/Bibos banteng) merupakan plasma nutfah nasional Indonesia, hasil domestikasi dari banteng liar beratus-ratus tahun yang lalu. Kelestarian sapi Bali di Indonesian semakin terancam karena sapi Bali merupakan pemasok daging utama di Indonesia Timur. Data tahun 2000 untuk pemenuhan kebutuhan ataupun eskpor, angka ini akan terus bertambah seiring dengan pertambahan penduduk. Jika dihitung dengan angka kematian maka akan terjadi penurunan jumlah populasi yang mencapai 26% dari total populasi. Hal ini menjadi masalah besar jika tidak diikuti dengan peningkatan populasi karena akan berdampak pada habisnya populasi sapi Bali galur asli Indonesia. Sapi Bali masih mempunyai sifat liar meskipun sudah di domestikasi. Penelitian yang mengungkap tentang profil hormon reproduksi khususnya estrogen pada sapi Bali masih sangat terbatas. Hal ini kemungkinan disebabkan karena masih adanya sifat liar pada sapi bali, sifat yang menjadi kendala di lapangan saat akan diterapkan aplikasi teknologi. Pendeteksian hormon ada 2 macam yaitu secara in- vasif dan non in- vasif, pengelompokan ini berdasarkan manipulasi pada hewan, pengambilan sampel dikelompokkan menjadi invasif (plasma/serum, saliva) maupun non- invasif (feses dan urin) pengambilan sampel secara invasif dipercaya dapat menjadi pemacu stres (stressor) bagi hewan namun metode ini masih banyak digunakan untuk pendeteksian

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hormon karena hasil pendeteksian merupakan gambaran pada saat pengambilan sampel. Keuntungan pada pengambilan dan pedeteksian secara non invasive adalah cara pengambilan sampel bukan menjadi stressor dan hewan tidak perlu dianestesi sedangkan kekurangan metode ini adalah kandungan hormon dalam feses atau urin merupakan metabolit sehingga tidak dapat menggambarkan kondisi fisiologi saat pengambilan sampel, selain itu pakan hewan kadang-kadang dapat mempengaruhi hasil. Belum adanya kajian mengenai profil hormon selama folikulogenesis menjadi kendala saat akan dilakukan aplikasi teknologi seperti inseminasi buatan (IB). Pemeriksaan kadar hormon estrogen dengan sampel secara metode invasif yang dipadukan dengan pemeriksaan hormon secara non invasif yang disertai pemeriksaan ovaria dengan menggunakan ultrasonografi merupakan introduksi baru dalam upaya pengembangan sapi Bali. Penelitian ini menggunakan 10 ekor sapi Bali, berumur 2 tahun, belum beranak dan sehat. Pada penelitian ini akan dilakukan pemeriksaan serial yaitu pemeriksaan diameter folikel yang disertai dengan pemeriksaan hormon secara invasif dan non invasif selama siklus estrus. Data hasil penelitian akan digunakan untuk menentukan panjang siklus, jumlah gelombang perkembangan folikel, dan profil hormon dalam darah dan feses. Pada penelitian ini digunakan 7 ekor dari 10 ekor sapi Bali yang ada di KP4 UGM Yogyakarta. Pemilihan sapi Bali yang digunakan berdasarkan kesiapan sapi Bali bereproduksi (minimal umur 2 tahun) serta hasil pengamatan siklus estrus secara

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eksternal yang didukung oleh peme riksaan menggunakan USG, sapi Bali yang bersiklus secara normal yang digunakan dalam penelitian ini. Berdasarkan hasil pengukuran diameter folikel didapatkan bahwa seluruh sapi Bali yang dikandangkan di KP4 UGM mempunyai 2 gelombang folikel. Identifikasi folikel dominan dapat dirunut mulai awal perkembangan yaitu folikel dengan diameter kecil sampai menjadi folikel yang berdiameter terbesar. Pada penelitian ini panjang siklus estrus sapi Bali 18.75 ± 1.02 hari, penentuan awal siklus ditandai dengan munculnya tanda-tanda estrus dengan lama waktu gelombang pertama 8.78 ± 1.60 hari dan gelombang 2 yaitu 9.78 ± 1.52 hari dengan diameter maksimal folikel pada gelombang 1 mencapai 6.71 ± 0.40 mm sedangkan diameter maksimal folikel pada gelombang 2 adalah 10.36 ± 0.41 mm. Pada penelitian ini kadar tertinggi estradiol pada folikel gelombang pertama yaitu 41.21 ± 5.6 pg/ml lebih rendah dibandingkan pada gelombang kedua kadar estrogen yang mencapai 70.95 ± 1.35 pg/ml Hal ini membuktkan bahwa level estradiol berkaitan dengan peningkatan ukuran diameter folikel. Kadar estradiol yang rendah pada gelombang folikel 1 ini yang kemungkinan menyebabkan terjadinya folikel atresia. Korpus luteum pada penelitian ini mencapai uk uran maksimal pada hari ke-13 ± 1.24 hari (kisaran 12-14 hari) dengan level hormon progesteron rata-rata 7.2 ± 1.07 ng/ml dan akan mulai regresi pada hari ke 14 dengan life span 9.64 ± 1.01 hari. Regresi korpus luteum merupakan aksi kerja hormon prostaglandin (PGF2a). Hormon PGF2a dihasilkan oleh uterus yang beraksi secara parakrin terhadap korpus

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luteum, mekanisme kerja parakrin inilah yang menyebabkan level PGF2a tidak dapat dideteksi secara invasif ataupun non invasif. Hasil uji hormon matabolit estradiol pada penelitian ini didapat bahwa adanya peningkatan nilai absorbensi seiring dengan tingginya pengenceran sampel, hal ini membuktikan level hormon estradiol menurun seiring dengan pengenceran sampel feses. Penurunan kadar estradiol pada sampel feses tidak mencapai 50%, hanya berkisar antara 20-30%. Pada penelitian ini hasil analisis sampel original konsentrasi sampel berada diantara 30-70% (50% = 92pg/ml) sehingga jika dilakukan pengukuran sampel berikutnya sampel tidak perlu dilakukan pengenceran atau jika dilakukan pengenceran maksimal 1:2. Hal ini bertujuan agar konsentrasi hormon yang diukur pada sampel berada pada garis linier dari kurva standar. Hasil uji paralelism terhadap el vel estron sampel feses menunjukkan adanya peningkatan nilai absorbensi seiring dengan pengenceran yang meningkat, hal ini berarti level hormon estron akan secara paralel menurun seiring dengan semakin tingginya dengan binding 50%. Penurunan nilai absorbensi estron tidak mencapai 50% hanya berkisar 20 % dengan nilai pengikatan estron 50% adalah 133 pg/ml. Penurunan nilai absorbensi yang tidak mencapai 50% ini bukan disebabkan karena kesalahan saat proses pengeringbekuan ataupun analisis karena nilai rata-rata recovery pada analisis ini 100% dan nilai R2 mencapai 0,99. Uji pararelism terhadap hormon progesteron sampel feses sapi Bali didapatkan bahwa adanya peningkatan nilai absorbensi seiring meningkatnya pengenceran dengan nilai pengikatan 50% adalah 1,76 ng/ml sehingga jika akan dilakukan

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pemeriksaan sampel feses, sampel hanya perlu diencerkan sampai 1:2. Pada penelitian ini level hormon progesteron dalam darah dan feses mempunyai kesamaan profil meskipun ada pergeseran hari saat level puncak progesteron. Perbedaan puncak level hormon steroid pada feses dan darah berkaitan dengan lama waktu metabolisme dan ekskresi feses. Hasil analisis korelasi pada penelitian ini menunjukkan adanya korelasi positif yang signifikan antara perkembangan folikel dengan level estradiol dalam sirkulasi selama siklus estrus. Hal ini menunjukan bahwa perkembangan folikel akan seiring dengan fluktuasi level estradiol. Hasil analisis korelasi pada penelitian ini menunjukan adanya korelasi postif antara diameter korpus luteum dengan level progesteron sapi Bali. Hasil dari semua tahap penelitian adalah dapat diteguhkan sapi Bali dara yang dikandangkan di KP4 UGM mempunyai 2 gelombang folikel dengan panjang siklus 18,57 ± 1,02. Level estradiol yang tertinggi dihasilkan pada folikel dominan pada gelombang folikel ke- 2 dalam satu siklus estrus yaitu rata-rata 70,95 ± 1,35 pg/ml. Level tertinggi hormon estradiol pada folikel gelombang pertama mencapai korpus luteum sapi Bali mulai terdeteksi pada hari ke- 3 setelah tanda-tanda estrus muncul dan dia meter tertinggi pada hari ke 13,21 ± 1,12 dengan level progesteron 6.89 ± 1.12 ng/ml. Berbeda dengan level estradiol dan progesteron dalam darah, level tertinggi hormon tersebut akan mengalami pergeseran hari. Level estradiol dalam feses akan mencapai puncak pada saat sapi Bali ovulasi dengan level 17,69 ± 1,77 ng/gr feses kering, sedangkan rata-rata level progesteron feses puncak 230,09 ± 42,13 ng/gr feses

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kering pada hari ke 14,3 ± 1,38. Hormon estradiol dan progesteron yang dihasilkan oleh sel-sel ovarium dan korpus luteum akan bersirkulasi menuju organ target, selain itu akan masuk ke dalam enterohepatic circulation yang pada akhirnya level hormon tersebut dapat terdeteksi di dalam feses. Penentuan panjang siklus estrus pada sapi Bali yang dikandangkan di KP4 dapat dilakukan berdasarkan perunutan ukuran folikel ovulasi, secara invasif dan non invasif.

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SUMMARY

Bali cattle (Bos sondaicus, Bos javanicus, Bos banteng, Bibos banteng) was national indigenous cattle of Indonesia, domestication result of wild cattle for hundreds years ago. Preservation of Bali cattle is threatened because they are usede as main meat supply in the eastern parts of Indonesia. In the year of 2000 to meet the needs or for exported, this rate will continually increase along with population growth. If it is calculated by mortality rate it will reach 26% of the total population. This becomes a big problem if not to be followed by population growth and will be impacted on the run out of Bali cattle population as Indonesia indigenous cattle. The Bali cattle still has a wild nature although it has been domesticated. The present research which reveals on profile of reproductive hormones especially estrogen and progeserone on Bali cattle was very limited. This was possibly caused by the wild nature of Bali cattle. This wild nature which becomes an obstacle in the field when technology will be applied. The approach of hormone determination has 2 kinds that are invasive and noninvasive methods. This grouping is based on manipulation on animal, sampling is grouped to be invasive (plasma/serum, saliva) or non- invasive (fesses and urine). Sampling invasively is believed can become stressor for animal but this method is still more used to detect hormone due to approaching result is a view when sampling is not to be stressor and animal necessarily to be anesthetized while the less of this method is hormone content in fesses or urine is metabolite so it cannot describe

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physiological condition when sampling, in additional animal feed sometimes can influence production. The lack of study in hormonal profiles for folliculogenesis becomes the obstacle when it will be conducted application of technology such as artificial insemination (AI), determination of estrogen hormone levels by sample and invasive method combined with hormonal checking non- invasively followed by ovarian checking using ultrasonography which is new introduction in the effort to develop Bali cattle farming system. The present research used 10 of Bali cattle heifers, 2 years old. It was conducted serial examinations, examination of follicular dynamics followed by examination of hormones with either invasive and non- invasive methods during estrous cycles. Result data will be used to determine the length of cycles, number of follicles developing, follicular waves, as well as blood and feces hormonal profiles. In the present research used 7 of 10 Bali cattle existing in the farm. The selection of Bali cattle used was based on the readiness of them for reproduction (minimum 2 years old) and observation result of estrous cycles

externally supported by

ultrasonographic examination. Bali cattle which cycling normally was used in this research. It was obtained that all Bali cattle kept in KP4 UGM had 2 follicular waves. Determination on a number of follicle waves uses USG (Honda, Japan) by measuring follicle diameter in serial. Identification of dominant follicle can be traced starting from initial development namely follicle with smallest diameter to become

the

biggest one. In this research, the length of estrous cycles in Bali cattle was 18.57 + 1.02 days, determination of initial cycles was signed by the rise of estrus signs with

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the long time of first wave is 8.78 ± 1.6 days and second wave is 9.78 ± 1.52 days with diame ter of maximal follicle on first wave was up to 6.71 ± 0.40 mm while the second one was 10.36 ± 0.41 mm. In the present research, the peak of estradiol levels in first wave follicle was 41.21 ± 5.6 pg/gr dry feces compared with the second wave in which levels of estrogen reach 70.95 ± 1.35 pg/gr dry feces. This fact proposed that estradiol level was related to the increase of folliculare diameter. Estradiol level which was low on the first follicle wave which possibly caused the follicle became atretic. Corpus luteum in the present research had maximum size on the day 13.00 ± 1.24 (around 12 - 14 days) with average level of progesterone was 7.20 ± 1.07 ng/ml and it will start regression

on

fourteenth day with life-span of 9.64 + 1.01 days.

Regression of corpus

luteum

was a result of

prostaglandin (PGF2α) hormone

action. Prostaglandin F2α was produced by the uterus. The mechanism of paracrine causing level of PGF2α can not be detected by invasive or non-invasive methods.. Test result of estradio l metabolite hormone in this research was obtained that there was an increase of absorbance rate in spontaneous with dilution of feces sample. Decrease of feces estradiol levels did not reach 50%, In this research, result analysis of original sample of sample concentration was between 30 and 70% (50% = 92 pg/ml) so it was conducted the measurement and was only between 20 and 30%. next sample, it was not necessary to be conducted dilution or if it was done so maximum

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dilution was 1 : 2. This was aimed in order that hormone concentration measured in the sample was located in linear line of standard curve. Test result of paralelism towards estrone level of feces sample indicated there was an increase of absorbance value in spontaneous with increasing dilution, this was meant that estrone hormone level will parallelly decreased in spontaneous with the highest of 15% binding. The decrease of estrone absorbance value did not reach 50%, it was only around 20% with estrone binding rate of 50% was 133 pg/ml. The Decrease of absorbance rate which did not reach 50% was not caused by the mistake when the process of drying and freezing or analysis due to average recovery value in this analysis was 100% and value of R2 reaches 0,99. Test of pararelism towards progesterone hormone of feces sample of Bali cattle was obtained that there was an increase of absorbance value in spontaneous with the increase of dilution with binding value of 50% was 1.76 ng/ml so if it will be conducted the examination of feces sample. It was only necessary to be diluted up to 1 : 2. In this research the blood and feces levels of progesterone hormone in had a similarity in profile although there was one day shift when it was on the highest level of progesterone. The difference of the highest level of steroid hormone in feces and blood related with long time of metabolism and feces excretion. Result analysis of correlation in this research indicated that there was a significantly positive correlation between the follicular development and the estradiol level in circulation for estrous cycles.

This fact indicated that the follicular

development will be spontaneous with fluctuation of estradiol level. Result analysis

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of correlation in this research indicates there is positive correlation between diameter of corpus luteum with progesterone level of Bali cattle. The result of all steps of research can be confirmed that Bali cattle caged kept in KP4 UGM had 2 follicular development waves with the length of estrous cycles was 18.57 + 1.02 days. The highest estradiol level was produced in dominant follicle on the second folliculare wave in one estrous cycles in average that was 70.95 + 1.30 pg/ml. Corpus luteum of Bali cattle started to be detected on the third day after estrus signs appear and largest diameter on day 13.42 + 1.01 with progesterone level was 7.20 ± 1.07 ng/ml. It is different with blood levels of estradiol and progesterone level, the hormone will experience a day shift. Estradiol level in feces will reach to the highest leve l when Bali cattles get an ovulation with the level of 17.69 ± 1.77 ng/gr dry feces, while average progesterone level of top fesses is 230.09 ± 42.13 ng/ gr dry feces on the day of 14.28 + 1.38. Estradiol and progesterone hormone produced by ovarium cells and corpus luteum will be circulated to lead to organ of target, in additional it will come into enterohepatic circulation finally the hormone level can be detected in fesses. Determination of the length of estrous cycles of Bali cattle kept in the farm can be done based on the decrease of ovulation follicle size, invasively and non invasively.

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