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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 pemeriksaan 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 ukuran 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 level 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 diameter 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 non-

invasive 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 diameter 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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DAFTAR PUSTAKA Achmad Nurchamdi., 2005., Karakteristik Sumberdaya Genetik Ternak Sapi Bali

(Bos-bibos banteng) dan Alternatif Pola Konservasinya., B I O D I V E R S I T A S Volume 6, Nomor 1 Januari 2005 Halaman: 70-75

Adams, G. P., R. L. Matteri, J. P. Kastelic, J.C.H. Ko, and O. J. Ginther. 1992.

Association between surges of follicle-stimulating hormone and the emergence of follicular waves in heifers. J. Reprod. Fertil. 94:177-188.

Adryani Ris ., I Ketut Suatha., I Wayan Batan., 2012., Keragaman Silak Tanduk Sapi

Bali Jantan dan Betina., Buletin Veteriner Udayana Vol. 4 No.2: 87-93 Airin., C.M., 2006., Korelasi estradiol dengan gambaran apus swab vagina pada

kambing peranakan ettawah selama siklus estrus., Thesis Pasca Sarjana Universitas Gadjah Mada.

Armstrong D.G and Webb R. 1997. Ovarian follicular dominance: the role of

intraovarian growth factors and novel proteins Reviews of Reproduction 2 139–146

Andre, L Niklaus, Graham W. Aberdeen, Jeffrey, S. B, Gerald J.P and Eugene D. A.,

2003. Effect of Estrogen on Vascular Endothelial Growth/ Permeability Factor Expression by Glandular Epithelial and Stroma Cells in the Babbon Endometrium. Biology of Reproduction.

Arimbawa I.W.P., Trilaksana I.G.N.B., dan Pemayun T.G.O. 2012. Gambaran

Hormon Progesteron Sapi Bali selama Satu Siklus Estrus. Indonesia Medicus Veterinus 2012 1(3) : 330 – 336

Asa et al., 2001

Asihara,Y dan Kasahara, Y. 2001. Immunoassay and immunochemistry. In John, B.H (eds), Clinical Diagnosis and Management by Laboratory Methods 21st ed..Philadelphia : WB Saunders Company.

Austin, E.J., Mihm, M., Evans, A.O.C., Ireland, J.L.H., Ireland, J.J., Roche, J.F.,

2002. Effects of oestradiol and progesterone on secretion of gonadotrophins and growth and differentiation of first wave follicles during the estrous cycle of beef heifers. Reproduction 124, 531–541.

|99

Austin.,Mihm. M., Evans., Knight. P.G., Ireland., J.LH., Roche., J.F., 2001., Alterations in intrafollicular regulatory factors and apoptosis during selection of follicles in the first follicular wave of the bovine estrous cycle. Biol. Reprod., 64: 839-848

Bahar, S. dan Rakhmat. 2003. Kajian pertumbuhan sapi bali yang digembalakan

dengan pakan hijauan lokal. Prosiding Seminar Nasional Teknologi Peternakan dan Veteriner. Bogor, 28-29 September 2003.

Bao B and Garverick HA., 1998., Expression of steroidogenic enzyme and

gonadotropin receptor genes in bovine follicles during ovarian follicular waves: a review., J Anim Sci. 1998 Jul;76(7):1903-21.

Bao B, Garverick HA, Smith GW, Smith MF, Salfen BE, Youngquist RS. 1997.,

Changes in messenger ribonucleic acid encoding luteinizing hormone receptor, cytochrome P450-side chain cleavage, and aromatase are associated with recruitment and selection of bovine ovarian follicles. Biol Reprod 56:1158-1168

Braden, TD., Gamboni F., and Niswender., 1988., Effects of Prostaglandin Induced

luteolysis on the populations cells in the ovine Corpus Luteum. Biol. Reprod. 39:245-253

Beg MA, Bergfelt DR, Kot K, Wiltbank MC, Ginther OJ. 2001. Follicular fluid

factors and granulosa-cell gene expression associated with follicle deviation in cattle. Biol Reprod; 64:432–441.

Beimborn, V. R., Tarpley, H. L., Bain, P. J., and Latimer, K. S., 2003. The Canine

Estrous Cycle: Staging Using Vaginal Cytological Examination. Ross University, School of Veterinary Medicine, St. Kitts, West Indies (Beimborn) and Department of Pathology, College of Veterinary Medicine, The University of Georgia, Athens, GA 30602-7388 (Tarpley, Bain, Latimer)

Bodlaender. 1995. Progesteron : Physiology and Clinical Utility. Melalui

www.dpcweb.com Borisenkow., 2000., The Content of Sex Hormones in The Enteric Medium Of

Female Ruminants., Journal of Evolutionary Biochmestry and Physiology., Vol 26, 1:45-49.

|100

Braw-Tal R, Yossefi S (1997). Studies in vivo and in vitro on the initiation of follicle growth in the bovine ovary. Journal of Reproduction and Fertility, 109: 165-171.

Brook CGD, and Marshall NJ.1996. Essential Endocrinology, 3rd Edition. Blackwell

Science. Brown, JL., Samuel KW., David ED., and Laura HG., 1994., Comparative Aspects of

Steroid Hormone Metabolism and Ovarian Activity in Felids, Measured Noninvasively in Feces'., BIOLOGY OF REPRODUCTION 51, 776-786

Chamidi AN., 2005., Karakteristik Sumberdaya Genetik Ternak Sapi Bali (Bos-bibos

banteng) dan Alternatif Pola Konservasinya., Biodiverditas. Volume 6, No 1: 70-75 ISSN: 1412-033X

Cullinan, K and Koos, R. D., 1993. Vascular Endothelial Growth Factor/Vascular

Permeability Factor Expression in Rat Uterus: Rapid Stimulation by Estrogen Correlates with Estrogen-Induced Increased in Uterine Capillary Permeability and Growth. Endokrinology 133:829-837

Cunningham, J, 2001. Textbook of Veterinary Physiology. 3rd ed. W. B Saunders

Company, Sydney. Cunningham, J, 2007. Textbook of Veterinary Physiology. 4th ed. W. B Saunders

Company, Sydney. Darmadja, S.D.N.D. 1980. Setengah Abad Peternakan Sapi Tradisional dalam

Ekosistem Pertanian di Bali. [Disertasi]. Bandung: Universitas Padjadjaran. Diskin MG., D.R Mackey, J.F Roche J.M Sreenan., 2003., Effects of Nutrition and

Metabolic Status on Circulating Hormones and Ovarian Follicle Development in Cattle., Animal Reproduction Science Volume 78, Issue 3 :345-370.

Ditjennak. 2008. Sapi bali. Edisi Februari2008. Http//www.ditjennak.go.id/

tgalerifotodetail. Entwistle, K. W., dan C. A. J. Ridd. 1995. Teknologi ELISA dalam diagnosis dan

penelitian : Asai hormon dengan ELISA. Editor G. W. Burgess. Gajah Mada University Press. Yogyakarta. (Diterjemahkan : W. T. Artama dan E. Moeljono)

Evan, and J. E. Fortune., 1997., Selection of the Dominant Follicle in Cattle Occurs in

the Absence of Differences in the Expression of Messenger Ribonucleic

|101

Acid for Gonadotropin Receptor, Journal Endocrinology Vol. 138, No. 7 2963-2971

Fattah, 2002 Forde, N., Beltman, M.E., Lonnergan, P., Diskin, M., Roche, J.E., Crowe, M,A.,

2011., Oestrous Cycle in Bos Taurus Cattle., Animal Reproduction Science. 124: 163-169

Frandson, R. D., 1996. Fisiologi dan Anatomi Ternak. Edisi Ke-4. Universitas

Gadjah Mada Press, Yogyakarta. Garverick, H.A., W.G. Zollers, and M.F. Smith 1992. Mechanisms associated with

corpus luteum lifespan in animals having normal or subnormal luteal function. Anim. Reprod. Sci. 28:111–124.

Gholib., 2011., Non Invasive Hormone Monitoring: Faecal Androgen and

Glucocorticoid In Male Crested Macaques (Macaca nigra) In Reation To Seasonal And Social Factors., Thesis Pasca sarjana IPB.

Ginther O. J. ,L. Knopf and J. P. Kastelic., 1989., Temporal Associations Among

Ovarian Events in Cattle During Oestrous Cycles With Two and Three Follicular Waves., J. Reprod. Fert. 89:223-230

Ginther OJ., 2000., Selection of the dominant follicle in cattle and horses. Anim

Reprod Sci 60–61:61-79 Ginther, O. J. and M. D. Utt. 2006. Doppler ultrasound in equine reproduction:

Principles, techniques, and potential. J. Equine Vet. Sci. 24:516–526. Ginther, O. J., L. Knopf, and J. P. Kastelic. 1989. Ovarian follicular dynamics in

heifers during early pregnancy. Biol. Reprod. 41:247–254. Ginther, O. J., M. A. Beg, F. X. Donadeu, and D. R. Bergfelt. 2003. Mechanism of

follicle deviation in monovular farm species. Anim. Reprod. Sci. 78:239–257.

Ginther, O.J., Bergfelt, D.R., Beg, M.A., Kot, K., 2002. Role of low circulating FSH

concentrations in controlling the interval to emergence of the subsequent follicular wave in cattle. Reproduction 124, 475–482

|102

Gordon DN, Jennifer L., Juengel , Patrick J, Silva M, Keith R., And Eric WM., 2000., Mechanism Contrlling the Function and Life Span of Corpus Luteum., Physiological Vol. 80. 1:1-27

Gosden RG, Hunter RHF, Telfer E, Torrance C, Brown N. Physiological factors

underlying the formation of ovarian follicular fluid. J Reprod Fertil. 1988;82:813–825.

Goymann W. 2005. Noninvasive monitoring of hormones in bird droppings:

physiological validation, sampling, extraction, sex differences, and the influence of diet on hormone metabolite levels. Ann NY Acad Sci 1046: 35–53.

Graham L. H., Goodrowe K. L., Raeside L., Liptrap R. W., 2005. Non-invasive

monitoring of ovarian function in several felid species by measurement of fecal estradiol-17ß and progestins

Gregory F. Erickson., 2008, The ovary Follicle Growth and Development., Glob. libr.

women's med. Gross, T.S. (1992). Development and use of faecal steroid analyses in several

carnivore species. In: W. Scaftenaar, R.M. and S.J. Dielman (Editors), The First International Symposium on Faecal Steroid Monitoring in Zoo Animals, Rotterdam, pp. 55-61.

Guyton AC, Hall JE, 1996. Textbook of Medical Physiology, 9th ed. WB, Saunders

Company, Philadelphia, Pennsylvania Hafez B and ESE Hafez., 2000., Reproduction in Farm Animals. Lippincont William

and Wilkins. A Wolter Kluwer Company. Philadelphia. Hamish MF., 2006., Regulation of The Ovarian Follicular Vasculatare., Reproductive

Biology and Endocrinology, 4:18 Hardjosubroto, W. 1994. Aplikasi Pemuliabiakan Ternak di Lapangan. Jakarta: PT.

Gramedia Widiasarana Indonesia. Heistermann M, Uhrigshardt J, Husung A, Kaumanns W and Hodges JK.2001.

Measurement of Fecal Steroid Metabilites in the lion-tail Macaque (Macaca silenus): a Non- invasif Tool for Assessing Female Ovarian Function. Primate report 59:27-42

|103

Hunter M.G., R.S., Robinson., G.E.Mann., Webb., R., 2004., Endocrine and Paracrine control of follicular development an ovulation rae in the farm species., Abimal Reproduction Science 83: 461-477

Hyder SM, Stancel GM, Chiappetta C, Murthy L, Boettger-Tong HL, Makela S. 1996.

Uterine expression of vascular endothelial growth factor is increased by estradiol as well as tamoxifen. Cancer Res, 56:3954-3960

Kaneko et al., 1997).

Kapit, W., Macey, I. R., Meisami, M. 1987. The Physiology Coloring Book.

AnImprint of Addison Wesley Longman, Inc. p: 141 Karuri, A. R., Kumar, A. M., and Mukhopadhyay, D., 1998. Different ial expression

and Selective Localization of vascular Permeability Factor/Vascular Endothelial Growth Factor in The Rat Uterus during The Oestrus Cycle. Journal of Endokrinology 159:489-49

Kastelic, J. P., L. Knopf, and O. J. Ginther. 1990. Effect of day of prostaglandin F2a

treatment on secretion and development of the ovulatory follicle in heifers. Anim. Reprod. Sci. 23:169-180.

Kastelic.J.P., 1999., Folliculogenesis in Cattle., Biotechnologio Reproducao En

Bovins (1st simposio Intercational De Reproducao Animal Aplcada) Knight PG, Glister C (2001). Potential local regulatory functions of inhibins, activins

and follistatin in the ovary. Reproduction, 121: 503-512. Kumar A, S Mehrotra, S S Dangi, G Singh, Subhash chand, Lakhanpal Singh, A S

Mahla, Sachin Kumar, K Nehra., 2013., Faecal steroid metabolites assay as a non- invasive monitoring of reproductive status in animals., Vet World 6(1):59-63

Kune dan Sholihati. 2007. Tampilan Berahi dan TingkatKesuburan Sapi Bali Timor

yang Diinseminasi.Jurnal Ilmu Ternak. 7 (1): 1 – 5 Kricka, L.J .1999. Principle of immunochernical technique. In Carl, A.B dan Edward,

R.A (eds), The textbook of'Clinical Chemistry. 3ed. 1999.Philadelphia WB. Saunders Company

|104

Lana, K., D. Djagra, dan K. Sulandra. 1979. Bobot lahir sapi Bali. Prosiding Seminar Nasional Penelitian dan Penunjang Peternakan. Denpasar,Bali: Universitas Udayana

Laurence and Shemesh. 2003. Naturally produced steroid hormones and their release

into the environment. Pure Appl. Chem., Vol. 75, Nos. 11–12, pp. 1859–1871

Litwack, G and Schmidt TJ. 2002.. Biochemistry of Hormones II: Steroid

Hormones,in: Textbook of Biochemistry and Clinical Correlations 5th ed.John Wiley and Sons.Pp 959-988

Lopez, H., R. Sartori, and M. C. Wiltbank. 2005. Reproductive hormones and

follicular growth during development of one or multiple dominant follicles in cattle. Biol. Reprod. 72:788–795

Lubis, A.M., dan P. Sitepu. 1998. Evaluasi Produktivitas Sapi Perah yang Terseleksi

di Dua Lokasi Penelitian KUD Sarwa Mukti dan KUD Pasir Jambu. Prosiding Seminar Nasional Peternakan dan Veteriner. Bogor.

Lussier JG, Matton P, Dufour JJ (1987). Growth rates of follicles in the ovary of the

cow. Journal of Reproduction and Fertility, 81: 301-307. Magoffin DA 2005 Ovarian theca cell. International Journal of Biochemistry & Cell

Biology 37 1344–1349. (doi:10.1016/j.biocel.2005.01.016) Mahmud Siswanto, Ni Wayan Patmawati, Ni Nyoman Trinayani, I Nengah Wandia, I

Ketut Puja., 2013., Penampilan Reproduksi Sapi Bali pada Peternakan Intensif di Instalasi Pembibitan Pulukan., Jurnal Ilmu dan Kesehatan Hewan., Vol. 1, No. 1: 11-15

Manna PR, Dyson MT & Stocco DM 2009 Regulation of the steroidogenic acute

regulatory protein gene expression: present and future perspectives. Molecular Human Reproduction 15 321–333. (doi:10.1093/ molehr/gap025)

McDonald, L.E., 2000. Veterinary Endocrinology and Reproduction. 3rd. Edition.

Bailliere Tindall, London. Mc Gee dan Hsueh. 2000. Initial and Cyclic Recruitment of Ovarian Follicles.

Endocrine Reviews 21(2): 200–214

|105

Mihm, M., Austin, E.J., Good, T.E.M., Ireland, J.L.H., Knight, P.G., Roche, J.F., Ireland, J.J., 2000. Identification of potential intrafollicular markers involved in selection of the dominant follicle in heifers. Biol. Reprod. 63, 811–819

Mihm, M., P. J. Baker, J. L. Ireland, G. W. Smith, P. M. Coussens, A. C. Evans, and

J. J. Ireland. 2006. Molecular evidence that growth of dominant follicles involves a reduction in follicle-stimulating hormone dependence and an increase in luteinizing hormone dependence in cattle. Biol. Reprod. 74:1051–1059.

Mostl, E., Choi, H.S., Wurm, W., Ismail, M.N. and Bamberg,. (1984). Pregnancy

diagnosis in cows and heifers by determination of oestradiol-17â in faeces. Br. Vet. J., 140

Monfort et al. 1998 Murray, K. M., 1996. The effect of esterogen and progesteron on structural changes

in the uterine glandular epitelium of the ovariectomized sheep. Biology of Reproduction 47. P: 408-417

Nasroallah Moradi and Kalsum., 2013., A Review of Biochemical Metabolites

Concentration Hormonal Composition of Folliculare Fluid in Domestic Animal., Annual Review and Resarch in Biology, 33:246-255

Noseir Wael MB., 2003., Ovarian Follicular Activity and Hormonal Profile During

Estrous Cycle in cCows: The Development of 2 Versus 3 Waves., Reproductive Biology and Endocrinology 1:1-6., http://www.RBEj.com/content/1/1/50

Nurlaili Palenga, 2012. Profill Progesteron Serum pada Sapi Bali Yang Diberi

Perlakuan Sinkronisasi Estrus Menggunakan Prostaglandin. Thesis Program Pasa Sarjana Sain Veteriner. Universitas Gadjah Masa Yogyakarta.

Oda, H, Y. Shiina, K. Seiki, N. Sato, N. Eguchi, and Y. Urade. 2002. Development and Evaluation of a Practical ELISA for Human Urinary Lipocalin-Type Prostaglandin D Synthase. Clinical Chemistry 48 (9): 1445–1453.

Oka, L. 2003, Performance of Bali cattle heifers and calves prior to weaning to a

feedlot system. Australian Centre for International Agricultural Reserch, Canberra

Palme . 2005. Measuring Fecal Steroids Guidelines for Practical Application., Ann.

N. Y. Acad. Sci. 1046:75-80

|106

Pane, I. 1990. Upaya peningkatan mutu genetik sapi Bali di P3 Bali. Prosiding

Seminar Nasional Sapi Bali. Bali, 20-22 September 1990. Parker KI, Robertson DM, Groome NP, Macmillan KL (2003). Plasma concentrations

of Inhibin A and follicle stimulating hormone differ between cows with two or three waves of ovarian follicular development in a single estrous cycle. Biology of Reproduction, 68: 822-828.

Payne WJA. Rollinson DHL. 1973 . Bali cattle . World Anim. Rev. 7:13-21 . Pedro H.H, Sarah Kamp, Scott Lusher and Gerrit H Veeneman. 2006. Non-steroidal

steroid receptor modulators. IDrugs 9(7):488-494 Pedro, 2009. The Textbook of Endocrinology Reproduction. Piccione G, Giovanni C, and Roberto R., 2003., Daily and Estrous Rhythmicity of

Body Temperature in Domestic Cattle., BMC Physiology, 3:1-8. http://www.biomedcentral.com/1472-6793/3/7

Prabowo, A., M. Sariubang, M. Sabrani, dan A. Tikupadang. 1992. Performans Sapi

bali Betina di Bawah Standar Bibit di Daerah Transmigrasi Kabupaten Luwu, Sulawesi Selatan. Gowa: Sub Balai Penelitian Ternak.

Prasodjo, Iis Arifianti, Kusdiantoro M., 2010. Korelasi Antar Lama Kebuntingan,

bobot Lahir dan Jenis Kelamin Pedet Hasil Inseminasi Buatan Pada Sapi Bali. Jurnal Veteriner Vol 11. No:1 : 41-45

Putri IN., 2014. Thesis Pasca Sarjana Fakultas Peternakan. Putro PP, Asmarani K, Sri G, Surya AP, Agung, B, Erif Maha NS, Slamet S, Claude

MA, Alfian N., 2012., Optimalisasi Pusat Pembibitan Pedesaan Village Breeding Center) Sapi Bali Sebagai Model Pengembangan Ternak Potong Dalam Menunjuang Program Swasembada Daging Sapi da Kerbau (PSDK) 2014 Di Kabupaten Barito Kuala. Laporan Kerjasama Penelitian Pengembangan Reproduksi Pemurnian Sapi Bali di Barito Kuala.

Putro PP., 2008., Dinamika Folikel Ovulasi dan Korpus lUteum Setelah Sinkronisasi

Estrus Pada Sapi Perah Peranakan Friesian Holstein. Disertasi Program Pasa Sarjana Sain Veteriner. Universitas Gadjah Masa Yogyakarta.

Putro, P. P. 2005. Gambaran reproduksi pada sapi perah rakyat. Seminar

Pengembangan Sapi Perah, Direktorat Perbibitan, Cisarua.

|107

Putro, P. P. 2008. Dinamika Folikel Ovulasi dan Korpus Luteum Setelah

Sinkronisasi Estrus pada Sapi Perah Peranakan Friesian Holstein. Disertasi S3, Program Pascasarjana Fakultas Kedokteran Hewan Universitas Gadjah Mada, Yogyakarta.

Quntara, R., Kopcow, L., Marconi, G., Sueldo, C., Speranza, G., and Baranao, R.I.,

2001. Relationship of Ovarian Stimulation Response with Vascular Endothelial Growth Factor and Degree of Granulosa Cell Apoptosis. Hum. Reprod.16:1814-1818

Rajesh J., 2007., Regulation of Follicular Wave Pattern in Cattle., Thesis.

Departement of Veterinary Biomedical Sciences, University of Sazkatchewan.

Raymond J and Helen F, 2010., Formation of The Follicullar antrum and Follicular

Fluid., Biology of reproduction, 82:1021-1029 Roche, J. F, 2004, Follicular waves in cattle, Veterinary Research Communication,

28: 107-110 Roche., 2000., Folliculare Waves in Cattle., Veterinary Reseach Communications

28:107-110 Ruswana Anwar, 2005., Sintesis, Fungsi dan Interpretasi Pemeriksaan Hormon

Reproduksi Disampaikan pada pertemuan Fertilitas Endokrinologi Reproduksi bagian Obstetri dan Ginekologi RSHS/FKUP Bandung.

Schwarzenberger, F., Mostl, E., Palme, R. Souza et al. [58] Bamberg, E., (1996).

Faecal steroid analysis for non- invasive monitoring of reproductive status in farm, wild and zoo animals. Anim. Reprod. Sci., 42: 515-526

Setyadi, A.M., 2011., Perubahan Alat Genitalia Eksterna pada sapi Bali dara selama siklus estrus di

Kebun Pendidikan Penelitian dan Pengembangan Pertanian (KP4) Universitas Gadjah Mada. Skripsi. Fakultas Kedokteran Hewan universitas Gadjah Mada

Sirois, J and Fortune, J.E. (1988) Ovarian follicular dynamics during the estrous cycle

in heifers monitored by real- time ultrasonography. Biol. Reprod. 39, 308-317.

Sturman et al (2000)

|108

Suryani W., 2012., Tingkah laku estrus sapi bali dara yang dikandangkan di kebun pendidikan, penelitian dan pengembangan pertanian (KP4) Universitas Gadjah Mada. Skripsi Fakultas Kedokteran Hewan, Universitas Gadjah Mada.

Talib, C., Siregar, A. R., Budiarti, T. and Diwyanto, K. 2003, Implementation of a

breeding program for Bali cattle. Australian Centre for International Agricultural Research, Canberra

Tarzillo and Fortune., 1993., Suppression of th secondary FSH surge with bovine

follicular fluid is associated with delayed ovarian follicular development in heifer. J. Reprod. Fertil 89: 643-653

Toelihere, M. R., 2003. Increasing the success rate and adoption of artificial

insemination for genetic improvement in Bali cattle. Australian Centre for International Agricultural Research, Canberra

Tongku et al (2010) Touma C, Palme R. 2005. Measuring fecal glucocorticoid metabolites in mammals

and birds: the importance of a biological validation. Ann NY Acad Sci 1046: 54–74.

Travis et al., (2002) Valdez et al Walsh SW, Mehta JP, McGettigan PA, Browne JA, Forde N, Alibrahim RM,

Mulligan FJ, Loftus B, Crowe MA, Matthews D, et al.: Effect of the metabolic environment at key stages of follicle development in cattle: focus on steroid biosynthesis. Physiol Genomics 2012, 44(9):504-517

Walsh., S, Jefferiss, CM, Stewart, K, Beresford., JN., 2003., IGF-I does not affect

the proliferation or early osteogenic differentiation of human marrow stromal cells., Bone Volume 33, Issue 1:80–89

Webb, R., G Arnsworthy, PC., Gong, J.G., Amstrong, D, G., 2004., Control of

Folliculare Growth: Lokal Interactions and nutritional Influences. J. Anim.Sci. 82:73-74

Williamson and Payne., 1978., Payne. 1978. Sapi Madura. Edisi Rabu,03

http://www.infoternak.com/sapibali.

|109

Young J M and A S McNeilly., 2010.,Theca: the forgotten cell of the ovarian follicle., Reproduction 140: 489–504