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ANIMAL PRODUCTION, September 2007, hlm. 123 128 ISSN 1411 2027 Terakreditasi No.

56/DIKTI/Kep/2005

Vol. 9 No.3

Non-Genetic Factors Influence on Doe Productivity Performance of Local Kejobong Goat under Village Production System
(Pengaruh Faktor Bukan Genetik terhadap Penampilan Produktivitas Induk Kambing Lokal Kejobong pada Sistim Produksi Pedesaan)
Akhmad Sodiq* and Budi Haryanto
Faculty of Animal Science, University of Jenderal Soedirman, Purwokerto

ABSTRAK: Penelitian bertujuan untuk mengetahui penampilan produktivitas induk kambing lokal Kejobong dan mengidentifikasi faktor-faktor lingkungan (bukan genetik) yang berpengaruh terhadap produktivitasnya. Penelitian lapang pada sistim produksi ternak ruminansia kecil melibatkan 212 ekor induk kambing. Koleksi data meliputi jumlah kepemilikan, identifikasi induk, jumlah cempe saat lahir dan sapih, jarak beranak, bobot sapih cempe, dan produktivitas induk. Prosedur General Linear Model (GLM) melalui bentuk fixed model diterapkan untuk menguji faktor tipe kelahiran dan paritas terhadap penampilan produktivitas induk. Hasil penelitian menunjukkan bahwa rataan jumlah cempe saat disapih 1,6 ekor, jarak beranak 268 hari, dan produktivitas induk 26,65 kg/induk/tahun. Faktor non genetik (tipe kelahiran dan paritas) nyata berpengaruh terhadap penampilan produktivitas induk kambing. Jumlah cempe saat disapih beserta produktivitas induk nyata meningkat hingga paritas keempat kemudian berangsur menurun kembali, dan juga meningkat pada tipe kelahiran kembar dua dan tiga. Jarak beranak nyata lebih pendek pada tipe kelahiran tunggal dibanding pada kelahiran kembar dua maupun tiga, demikian pula peningkatan paritas nyata memperpendek jarak beranak. Peningkatkan produktivitas induk kambing direkomendasikan melalui usaha perbaikan pengelolaan, utamanya adalah memperpendek jarak beranak dan meningkatkan jumlah cempe hidup hingga disapih. Kata Kunci: Produktivitas induk kambing, jarak beranak, jumlah cempe saat disapih, tipe kelahiran, paritas

Introduction
The majority of goats in Indonesia are concentrated on the Island of Java, and 23% of total population located in Central Java (DGLS, 2006). The major breeds being the Kacang and Peranakan Etawah goat (Djajanegara and Setiadi, 1991). Kacang is an indigenous breed of goat found in Indonesia. Peranakan Etawah goats are descended originally from crossing between the Kacang and Etawah goats. Local Kejobong goats are widely distributed over the whole areas of 13 villages at Kejobong Subdistric with the total population 15.317 heads. They have a relatively medium body frame and hanging ears, a convex face and horns in both sexes. Most animals are black, white and brown, although the pattern of colour is not necessarily uniform. Goats are kept primarily for meat production, so production traits of interest are the number of young weaned and the total weaning weight per doe per year.

Corresponding author: e-mail: akhmad_sodiq@yahoo.com

Increasing the productivity of goats will enhance national development planning for increasing rural income, reducing poverty and also increasing the level of protein consumption (Sodiq, 2005; Sodiq and Tawfik, 2003; Bradford, 1993). Key production traits considered for improving productivity in meat goats are adaptability and productivity conditions, reproductive rate, growth rate and carcass value (Luginbuhl, 2002). The level of productive performance is dependent on the interaction of genetic and environmental factors (Ahmadu and Lovelace, 2002; Gnes et al., 2002; Greyling, 2000), and also affected by feeding (Akingbade et al., 2004; Muir, 2006; Lassoued et al., 2006) and management system (Dickson-Urdaneta, et al., 2000; Song et al., 2006). Meat production in animals is affected by such variables as growth, weight at different ages, mortality, parturition interval, milk yield and mothering ability (Awemu et al., 2002). Due to the slow growth rates and long kidding intervals the flock productivity in terms of weaned live kid weight (kg) per doe per year was low (Ndlovu and Simela, 1996). High mortality of young stock and poor reproductive efficiency of dams are major causes of

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low productivity in many livestock production system. Low productivity of small ruminants in the tropics to poor management which predisposed the animals to diseases. Productivity rates are therefore generally low, with low kidding percentages and high mortality. The interval between kiddings is an important predictor of lifetime productivity. Wilson et al. (1985) have shown that prolonged kidding interval were responsibility for a decrease in overall productivity of goats. Kidding interval itself has been reported to be affected by a number of environmental factors (Wilson and Light, 1986, Mtenga et al., 1994). The objectives of this study are to find out the level of does productivity, and to assess the influence of environmental (non-genetic) factors on doe productivity of local Kejobong goats under village production system.

model. Independent variables in the model were type of birth and parity. The number of livestock in a flock (converted to the Livestock Unit of smallruminants) as a source of variance was absorbed in the calculation of statistical analysis. Because in unbalanced models most interactions are meaningless in order to better estimate the effects of the principals factors a model without interactions was finally adopted.

Results and Discussion


Litter Size at Weaning
Average litter size at weaning (Table 1) were higher than values recorded for Peranakan Etawah goat breeds (Sodiq, 2004). Litter size at birth and parity significantly influenced litter size at weaning (P<0.01). Litter size at weaning increased with parity, with the largest litters at the fourth parity and then slightly decreased. These results are consistence with those of Wilson and Light (1986) and Sodiq (2004). This may be attributed to physiological maturity of older does and their ability to provide enough milk for the kids. Increasing in litter size at weaning with parity due to an increased rate of twinning as parity increased. The number of litter at weaning depends on the doe ability in term of the survivability their kids. Previous researchers, Husain et al. (1995), Okello (1993), Alexandre et al. (1999), Madibela et al. (2002), Sodiq (2004), and Hailu et al. (2006) reported that survival rate until weaning was significantly affected by type of birth. Awemu et al. (2002) working on Red Sokoto goats found that the mortality tended to decrease as parity increased. Survival rate till weaning tended to decrease with larger size litters which also agree with the observations of Awemu et al. (1999), Ameh et al. (2002), and Mtenga et al. (1994). Kids from multiple births are often weak at birth as a result of physiological starvation in the uterus and lower energy reserves. Sodiq (2004) working on Kacang and Peranakan Etawah goats revealed that survival rate till weaning increased with the advance in parity up to the 4 th parity and then slightly decreased. This may be attributed to the physiological maturity of older does and their ability to provide enough milk for the kids.

Research Methods
Animal and Management
The study was conducted at small-ruminants smallholders over all 13 villages of Kejobong subdistric, Purbalingga regency, Central Java. All locations relatively have similar environmental conditions. Forage availability follows the distribution of rainfall with abundant and good quality forages in the rainy season. In the wet season, forages is generally scarce and of low quality. Local goats, mostly distributed and found in the Kejobong area of Purbalingga, is characterized by a relatively medium in body frame and hanging ears, a convex face and horns in both sexes. Most of the animals are black (74.50%), white (13%), and brown (12.5%). Management of animals was intensive by cut and carry feeding system and natural mating method. Mostly, a supplementary concentrate was not fed to the animals. Animals were provided with stilted housing in small flock (less than 10 heads). The purpose of raising goat is mostly for meat production.

Data Collection and Analysis


The following data were recorded, flock size, identification of does, litter size at birth and weaning, interval between kiddings, weaning weight, and doe productivity. The General Linear Model (GLM) procedure of the Statistical Package for Service Solution were used (SPSS, Inc. 1999 a,b ). Data on litter size at weaning, kidding interval and doe productivity were analyzed using a mixed

Kidding Interval
The least squares mean for kiddings intervals was 268 days (Table 1) ranging from 210 to 260

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days and varying according to type of birth and parity. This value is much larger than the 242 days reported for Kacang goats (Sodiq, 2004). Average kidding intervals of local Kejobong goat (Table 1) were close to values reported by Amsar et al. (1992). In this study (Table 1) demonstrated that parity significantly affected the interval between kiddings which generally decreased in consonance with the report of Wilson and Light (1986) that females at earlier parities take longer than older ones to return to reproductive status. Sodiq (2004) reported that parity significantly affected the kidding interval of Kacang and Peranakan Etawah goats which generally decreased with parity until the 4 th parity. Results of Odubote (1996, 2000) studies on West African Dwarf demonstrated that there was a significant decrease in the kidding interval from the 4 th parity. These results are also consistent with the report of Wilson and Light (1986) that females at earlier parities take longer than older ones to return to reproductive status. Das (1993) demonstrated that old does (3-4 year) tended to have lower kidding intervals than the younger (1-2 years) and older does (>5 years). This is probably due to the reproductive physiology function being more active in 3-4 years old does compared to lower activity in younger and older does. Kidding interval was significantly (P<0.01) affected by the type of birth (Table 1). The same results were also reported by Christopher (2001), Akusu and Ajala (2000), Greyling (2000), and ztrk and Akta (1996). These results (Table 1) demonstrated that the averages of the kidding intervals shortened progressively with the advance in the type of birth. Results of Christopher (2001)

found that the does with multiple births tend to have a shorter gestation length with 1 to 2 days difference between twins and triplets. Amoah et al. (1996) reported that the gestation period decreased as the litter size of the doe increased (b = -0.92 d/kid). Akusu and Ajala (2000) investigated on West African Dwarf goats concluded that does with single kids have a longer gestation than those with twins and triplets. Other researchers, Maria and Ascaso (1999) reported that the longer interval could be due to their larger litters. Milk yield of ewes lambing twins ranges from 1.7 to 1.8 kg and increase to 2.42.5 kg for those lambing triplets. A high milk production could lead to a negative energy balance. The magnitude of energy deficiency seems to affect these processes of follicular growth and development leading to the first ovulation (Nett, 1987).

Doe Productivity
The overall doe productivity of local goat at Kejobong Purbalingga was 26.650.78 kg/doe/year. The values reported in this study are higher than those reported by Anggraeni et al. (1995), and Sutama (1995). Zhou et al. (2003), Bearden and Fuquay (2000), and Ingo (1999, 2002) demonstrated that the environmental factors exerted a significant influence on the productivity. This results (Table 1) revealed that doe productivity of local Kejobong goat was significantly (P<0.01) affected by parity. Greyling (2000) and Marai et al. (2002) reported that the productive performance is dependent on the interaction of genetic and environmental factors and the effects of parity were significant. Results of

Table 1. Least squares means (LSM) and standard error (SE) for litter size at weaning, kidding interval and doe productivity Litter size at weaning Kidding interval Doe productivity Traits n (head) (days) (kg/doe/year) LSM SE LSM SE LSM SE
Overall Parity Parity 1 Parity 2 Parity 3 Parity 4 Parity 5 Parity 6 Parity 7 Type of birth Single Twin Triplet
a,b,c,

212 78 53 44 13 11 4 9 66 128 18

1.60 ** 1.46 a 1.62 a 1.75 bc 1.92b 1.82 bc 1.50 ac 1.33 ac ** 0.99 a 1.86 bc 2.06 c

0.04 0.07 0.09 0.08 0.14 0.12 0.29 0.17 0.01 0.03 0.04

268 * 278a 269a 261 b 256 b 256 b 255 b 253 b ** 296a 259 b 237c

2.45 4.68 5.01 4.43 7.29 8.45 9.36 5.27 4.49 2.44 4.78

26.65 ** 23.59 a 26.87 a 29.75 b 32.69 b 31.31 b 25.34 a 22.89 a ** 14.63 a 31.27 b 37.87 c

0.78 1.31 1.57 1.75 2.83 2.58 4.43 2.98 0.32 0.64 4.41

Means in the same column with different superscripts are significantly different (P<0.05) * P<0.05, ** P<0.01

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Ndlovu and Simela (1996) showed that due to the slow growth rates and long kidding intervals the flock productivity in terms of weaned live kid weight (kg) per doe per year was low. Awemu et al. (2002) reported that the parity significantly influenced doe productivity. Table 1 demonstrated that the average of doe productivity increased with the advance in parity up to the 4 th parity and slightly decreased thereafter. Results of Steve and Marco (2001) showed that the goat productivity may be positively correlated with maternal age, but decreased slightly after 9 years of age. It was indicated that the relationship between age and kid production was curvilinear. Awemu et al. (2002) investigation on Red Sakoto goats found that the doe productivity (kg/doe/year) in parity 1, 2, 3, 4, 5 and 6 were 20.9, 21.4, 22.5, 23.6, 27.9 and 33.4 kg, respectively. There was a significant influence (P<0.01) of type of birth on doe productivity of local Kejobong goat (Table 1). Similar results also were reported by Awemu et al. (2002) that the type of birth of goat significantly influenced the productivity. These results (Table 1) demonstrated that average of doe productivity increased progressively with the advance in type of birth. Awemu et al. (1999) reported that increasing litter size at birth and at weaning can improve the productivity of goats. Awemu et al. (2002) found that the effect of type of birth was highly substantial in goats, with quadruplets births producing 32.8 kg more meat at weaning than single births. The productivity of goat depends on the number of litter at birth, survival rate till weaning and interval between kiddings (Sutama, 1995). The effect of the type of birth was highly substantial in goats, with multiple births producing more than single births and the prolonged kiddings interval was responsible for a decrease in reproduction and productivity of goats (Awemu et al., 1999). The interval between parturition and the first post partum estrus is an important trait which contributes to the production efficiency (Greyling, 2000).

doe productivity were 1.600.04 heads, 2682.45 days, and 26.650.78 kg/doe/year, respectively. Litter size at weaning and doe productivity increased with parity, with the largest values at the fourth parity and then slightly decreased thereafter. Litter size at weaning and doe productivity also increased progressively with the advance in the type of birth. Kidding intervals shortened progressively with the advance in the parity and the type of birth. The results call for management effort to reduce intervals between kiddings and improve litter size at weaning, in order to improve the doe productivity of local Kejobong goat under village production system.

Acknowledgement
The authors wish to thank the small-ruminants smallholders at Kejobong Purbalingga. The study was supported by a competitive grant of Jenderal Soedirman University.

References
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Conclusion
The study has revealed that the non-genetic (type of birth and parity) exerted significant influences on doe productivity performance. Average litter sizes at weaning, kidding interval and

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Amoah, E.A., S. Gelaye, P. Guthrie and C.E. Rexroad, 1996. Breeding season and aspects of reproduction of female goats. Journal Animal Science 74: 723728. Amsar, Soedjadi, A. Priyono dan A. Setyaningrum, 1992. Growth and preweaning survivability of goats in Cilacap. Research Report. University of Jenderal Soedirman. Purwokerto. Pp.42. Anggraeni, D., R.S.G. Sianturi, E. Handiwirawan and B. Setiadi, 1995. The effect of improving management system on doe and ewe productivity. Proceedings Sciences and Technology National Meeting. Bogor, Indonesia. 25-26 January, 1995. Pp.374-379. Awemu, E.M., L.N. Nwakolar and B.Y. Abubakar, 1999. Environmental influences on pre-weaning mortality and reproductive performance of Red Sakoto does. Small Ruminant Research 34: 161-165. Awemu, E.M., L.N. Nwakalo and B.Y. Abubakar, 2002. The biological productivity of the Yankasa sheep and the Red Sakoto goat in Nigeria. Dept.of Animal Science, University of Nigeria, Nsukka, Nigeria. Bearden, H.J. and J.W. Fuquay, 2000. Applied Animal Reproduction. 5 th. Edition. Prentice Hall, Inc. Upper Saddle River. Pp.382. Bradford, G.E., 1993. Small ruminant breeding strategies for Indonesia. Proceedings of a Workshop held at the Research Institute for Animal Production. Bogor, August 3-4, 1993. Pp.83-94. Christopher, D.L., 2001. Boer Goat production: progress and perspective. Proceedings of the 2001 Conference on Boer goats. Beijing, China, Oct. 20 25, 2001. Das, S.M., 1993. Reproductive parameters and productivity indices of Blended goats at Malya Tanzania. International Foundation for Science Workshop Animal Production Scientific. Workshop for East African IFS. Kampala, Uganda, April, 1922, 1993.

Greyling, J.P.C., 2000. Reproduction traits in the Boer goat doe. Small Ruminant Research 36: 171-177. Gnes, H., P. Horst, M. Evrim and A. Valle-Zrate, 2002. Studies on improvement of the productivity of Turkish Angora goats by crossing with South African Angora goats. Small Ruminants Research 45(2): 115-122. Hailu, D., G. Mieso, A. Nigatu, D. Fufa and D. Gamada, 2006. The effect of environmental factors on preweaning survival rate of Borana and Arsi-Bale kids. Small Ruminant Research 6(1-3): 291-294. Husain, S.S., P. Horst and A.B.M.M. Islam, 1995. Effect of different factors on preweaning survivability of Black Bengal Kids. Small Ruminant Research 18: 1-5. Ingo, H., 2002. Analysis of survival curves in seasonally mated pastoral goat herds in northern Kenya using logistic regression techniques. Journal of Arid Environments 50: 621-640. Ingo, H., 1999. Effect of seasonality on the productivity of pastoral goat herds in Northern Kenya. [Dissertation]. Humboldt University, Berlin. Pp.181. Lassoued, N., M. Rekik and H. Ben Salem, 2006. Dargouth Reproductive and productivity traits of goats grazing Acacia cyanophylla Lindl. with and without daily PEG supplementation. Livestock Science, 105(1-3): 129-136. Luginbuhl, J.M. 2002. Breeds of goats for meat goat production and production traits. Department of Animal Science, NCSU. http://www.cals.ncsu.edu/an_sci/extension/animal/ meatgoat [20 Agustus 2002]. Madibela, O.R., B.M. Mosimanyana, W.S. Boitumelo and T.D. Pelaelo, 2002. Effect of supplementation on reproduction of wet season kidding Tswana goats. South African Journal of Animal 32(1): 122. Marai, I.F.M., E.I. Abou-Fandoud, A.H. Daader and A.A. Abu-Ella, 2002. Reproductive doe traits of the Nubian (Zaraibi) goats in Egypt. Small Ruminant Research 46: 201-205. Maria, G.A. and M.S. Ascaso, 1999. Litter size, lambing interval and lamb mortality of Salz, Rasa Aragonesa, Romonov and F1 ewes on accelerated lambing management. Small Ruminants Research 33: 167-172. Mtenga, L.A., G.C. Kifaro and B. Belay, 1994. Studies on factors affecting reproductive performance and mortality rates of Small East African goats and

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Sodiq, A., 2005. Small ruminants: implication and research strategies on rural poverty reduction. Journal of Rural Development 1(7): 1-7. Sodiq, A. and E.Z. Tawfik, 2003. The Role and breeds, management systems, productivity and development strategies of goats in Indonesia. Journal of Agriculture and Rural Development in the Tropics and Subtropics 104(1): 71-89. Song, H.B., I.H.Jo. and H.S. Sol, 2006. Reproductive performance of Korean native goats under natural and intensive conditions. Small Ruminant Research 65(3): 284-287. SPSS Inc., 1999a. SPSS for Windows: Base Systems Users's Guide Release 10.0. Michigan Avenue, Chicago. SPSS Inc., 1999b. SPSS Advanced Model 10.0. South Wacker Drive, Chicago. Pp.333. Steve, D.C. and F.B. Marco, 2001. Reproductive success in female mountain goats: the influence of age and social rank. Animal Behaviour 62: 173-181. Sutama, I.K., 1995. Puberty and early reproductive performance of Peranakan Etawah goat. Proceedings of the 7 th AAAP Animal Science Congress, Bali, July 11-16, 1994. Wilson, R.T. and D. Light, 1986. Livestock production in Central Mali. Journal of Animal Science (6): 557567. Wilson, R.T., C.P. Peacock and A.R. Sayers, 1985. Preweaning mortality and productivity indices for goat and sheep on Masai ranch in Kenya. Animal Production (41): 201-206. Zhou, H.M., D.Allain, J.Q. Li, W.G. Zhang and X.C. Yu, 2003. Effects of non-genetic factors on production traits of Inner Mongolia cashmere goats in China. Small Ruminant Research 47: 85-89.

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Vol. 9 No.3

Efek Pemberian Prostaglandin F2, secara Intra Vaginal Spons (IVS) dan Intra Muskuler (IM) terhadap Peningkatan Kinerja Reproduksi Domba
(The Application of Synchronization Methods Using Prostaglandin F2, by Intra Vaginal Sponges (IVS) and Intra Muscular (IM) to Improve Reproductive Performance of Thin tailed Ewe Lambs)
Rosmawati Saoeni
Balai Besar Pelatihan Kesehatan Hewan Cinagara, Bogor

ABSTRACT: The purpose of the experiment was to investigate the effect of different administration method of PGF2, i.e. intra vaginal sponges (IVS) and intra muscular (IM) on the onset and the duration of estrus, and Non-return Rate (NR) in thin tailed ewe lambs. A total of 20 thin tailed ewe lambs, aged 12-15 months, were at random assigned to one of four treatment groups in Completely Randomized Designed (CRD) : Animals in Treatment group I (P 1) received PGF2 of 5 mg/ml/head intramuscularly; Treatment group II (P2) received PGF2 of 5 mg/ml/head by intra vaginal sponges for twodays; Treatment group III (P3) received PGF2 of 5 mg/ml/head by intra vaginal sponges for four days; Treatment group IV (P4) received PGF2 of 5 mg/ml/head by intra vaginal sponges for six days. Two rams, aged 2-2.5 years used as a mated. Each treatment was repeated five times. Variables measured were onset and duration of estrus, and Non-return Rate (NR) in 30 days. Collected data were analyzed using analysis of variance followed by Post-hoc of Least Significant Difference (LSD). Average values of onset of estrus for P1, P2, P3 and P4 were 22.91, 23.16, 26.31 and 44.57 hours, respectively. Average values of duration of estrus for P 1, P2, P3 and P4 were 26.36, 48.36, 94.65 and 146.56 hours, respectively. Analysis of variance indicated that the administration method of PGF2 affected significantly (P<0.01) on the onset and the duration of estrus. Non-return Rate (NR) in 30 days for P1, P2 , P3 and P4 was 100,100, 20 and 20 percent, respectively. In conclusion, the application of estrous induction methods using Prostaglandin F2 by intra vaginal sponges (IVS) for two days and intra muscular (IM) can improve reproductive parameters of thin tailed ewe lambs. Key Words: Prostaglandin estrous, non-return rate, sheep

Pendahuluan
Ternak domba di Indonesia mempunyai prospek yang baik untuk dikembangkan, baik secara komersial maupun secara sambilan, dan cukup menguntungkan bagi petani kecil maupun buruh tani. Hal tersebut karena ternak domba memiliki sifat karakteristik reproduksi yang tinggi, dengan jumlah anak satu sampai empat ekor per induk untuk setiap kelahiran. Hampir semua breed domba sanggup menghasilkan lebih dari satu ekor anak per kelahiran, namun rata-rata literr size (jumlah anak per kelahiran) hampir selalu kurang dari dua, sehingga dapat dikatakan kemampuan reproduksi belum optimal (Hunter, 1980). Keadaan ini dapat ditanggulangi dengan perangsangan ovulasi untuk meningkatkan jumlah ovum per berahi melalui metode natural misalnya dengan meningkatkan kualitas pakan atau penggunaan pejantan (Evans dan

Maxwell, 1987) dan metode hormonal misalnya dengan pemberian hormon gonadotropin (FSH, follicle stimulating hormone dan LH, luteinizing hormone) atau gabungan keduanya (Hafez, 1987). Gertak berahi merupakan salah satu upaya untuk meningkatkan kinerja reproduksi domba agar dicapai jarak beranak yang optimal, sehingga pada akhirnya produksi anak (cempe) menjadi optimal. Hormon luteolitik yang umum digunakan untuk gertak berahi adalah prostaglandin F2 (Sumaryadi, 2003). Dasar fisiologis dari gertak berahi adalah hambatan pelepasan FSH dari hipofisis anterior sehingga menghambat pematangan folikel de Graaf atau penyingkiran corpus luteum (CL) baik secara manual maupun secara fisiologis. Prostaglandin F2 merupakan preparat hormon luteolitik yang berfungsi menginduksi kejadian berahi melalui penyingkiran CL. Data yang ada di Kabupaten Bogor, menyebutkan bahwa perkembangan populasi ternak

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domba hanya mampu mencapai kenaikan sebesar 0,56% per tahun dari target yang diharapkan sebesar 2% per tahun. Upaya untuk mencapai target tersebut antara lain dengan meningkatkan kinerja reproduksi melalui seleksi bibit, gertak berahi, superovulasi, IB, dan perbaikan manajemen pemeliharaan. Terkait dengan hal tersebut pemberian Prostaglandin F2 pada domba betina ekor tipis melalui teknik yang berbeda intra vagina spons (IVS) dan intra muskuler (IM) perlu dikaji terhadap tingkat kinerja reproduksinya (onset berahi, lama berahi, non return rate).

Metode Penelitian
Materi Penelitian
Materi yang digunakan dalam penelitian adalah : a. Dua puluh (20) ekor domba betina ekor tipis yang terseleksi, berumur 12 15 bulan, kisaran bobot badan 23 26 kg dan dua (2) ekor domba pejantan terseleksi umur 2 2,5 tahun. b.Hormon prostaglandin F2 yang digunakan empat (4) flakon (tiap 1 ml mengandung 5 mg prostaglandin F2) dengan merk Lutelyse produk dari Up John, serta minyak pelicin (minyak non kolestrol), kapas, kertas lakmus pH 6.0-7.0, tissue c.Holder untuk memasukkan spons dan alat suntik serta pengukur waktu, mikroskop, gelas obyek, gelas penutup, pipet pasteur, ose, tabung sperma.

b. Semua domba percobaan di adaptasikan selama dua minggu untuk menyeragamkan kondisi pakan dan lingkungan percobaan. c. Melakukan tes kualitas semen domba jantan yang akan digunakan untuk mengawini. d. Pengelompokan domba percobaan secara acak berdasarkan 4 macam perlakuan teknik pemberian prostaglandin F2 sesuai dengan yang ditetapkan. Teknik IM dilakukan dengan menyuntikan hormon prostaglandin F2 sebanyak 5 mg/ml/ekor ke dalam otot. Teknik IVS dilakukan dengan memasukan spons yang mengandung hormon prostaglandin F2 5 mg/ml/ekor ke dalam vagina masing-masing selama 2, 4, dan 6 hari. Pada kelompok perlakuan IM, domba percobaan diamati onset berahi, lama berahi. e. Melakukan pengamatan berahi setiap jam setelah penyuntikan dan pencabutan spons sampai domba percobaan menunjukkan tanda-tanda berahi. f. Melakukan perkawinan alam domba-domba yang berahi dengan pejantan yang telah dipersiapkan dengan rasio 1:10. g. Pengamatan non return rate (NR) 30 dari masingmasing perlakuan.

Peubah yang diamati adalah Onset Berahi


Cara menentukan onset berahi adalah dengan mengamati domba yang berahi setelah diberi perlakuan prostaglandin F2 secara IVS dan IM. Waktu onset berahi dihitung dari mulai disuntik hormon prostaglandin F2 sebanyak 5 mg/ml/ekor atau pemasukan spons yang mengandung hormon prostaglandin F2 sebanyak 5 mg/ml/ekor sampai munculnya awal berahi dalam satuan jam. Yang ditandai dengan, sering mengembik-embik, sering kencing sedikit seolah terputus-putus, mengangkat atau menggerak-gerakkan ekor sehingga mulai terlihat pembengkakan vulva. Hal tersebut di atas sesuai dengan pendapat (Mc. Donald, 1975; Toelihere, 2003; dan Tomaszewska et al., 1991).

Rancangan Penelitian
Metode penelitian yang digunakan adalah metode eksperimental, menggunakan Rancangan Acak Lengkap (RAL). Perlakuan terdiri dari empat (4) macam yaitu: P 1 : Pemberian prostaglandin F2 5 mg/ml/ekor secara intra muskuler sebagai pembanding. P 2 : Pemberian prostaglandin F2 5 mg/ml/ekor secara intra vagina spons selama 2 hari. P 3 : Pemberian prostaglandin F2 5 mg/ml/ekor secara intra vagina spons selama 4 hari. P 4 : Pemberian prostaglandin F2 5 mg/ml/ekor secara intra vagina spons selama 6 hari. Masing-masing perlakuan diulang 5 kali, sehingga dibutuhkan 20 ekor domba betina ekor tipis.

Lama Berahi
Lama berahi pada domba ditentukan dengan pengamatan domba yang mengalami berahi sejak awal timbulnya berahi sampai dengan gejala berahi hilang dalam satuan jam. Hasil pengamatan menunjukan tanda-tanda berakhirnya berahi adalah tidak mengeluarkan lendir lagi di vulva, tidak gelisah, vulva tidak membengkak dan mulai kuncup tidak mau dinaiki pejantan (Ditjenak, 1983).

Prosedur Kerja Penelitian


a. Melakukan identifikasi ternak domba percobaan sesuai dengan kriteria koefisien teknis yang homogen.

Efek Pemberian Prostaglandin F2 (Saoeni)

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Non Return Rate (NR) 30 hari


Menentukan kebuntingan seekor hewan dengan NR adalah mengamati berahi hewan yang bersangkutan mulai dari dikawinkan sampai dengan hari ke 30. Hal tersebut sesuai dengan pendapat Hunter (1980) bahwa setelah perkawinan domba percobaan diamati lagi kondisi berahinya. Jika ternak domba tidak menunjukkan berahi kembali selama 30 hari maka dianggap bunting, sebaliknya jika terjadi kembali berahi maka dianggap tidak bunting.

Analisis Data
Data onset dan lama berahi dianalisis menggunakan analisis ragam, jika terjadi perbedaan antar perlakuan dilanjutkan dengan uji Beda Nyata Terkecil (BNT), sedangkan data NR dianalisis secara deskriptif.

Hasil dan Pembahasan


Onset Berahi
Rata-rata onset berahi domba ekor tipis yang diberi prostaglandin F2 secara IM dan IVS dengan durasi yang berbeda secara rinci disajikan pada Tabel 1.
Tabel 1. Rataan onset berahi (jam) pada domba Perlakuan P 1 (IM) P 2 (IVS 2 hari) P 3 (IVS 4 hari) P 4 (IVS 6 hari)
abc

Rata-rata 22,91 0,77 a 23,16 1,51 a 26,31 0,08 b 44,57 1,09 c

Superskrip yang berbeda pada kolom yang sama menunjukan ada perbedaan pada P<0,01

Berdasarkan Tabel 1 diketahui bahwa onset berahi pada domba ekor tipis berkisar antara 21,15 45,49 jam dengan rata-rata 29,24 jam, setelah di berikan hormon prostaglandin F2 secara IM maupun secara IVS. Mayoritas (75%) domba menunjukkan onset berahi antara 21,15 - 26,37 jam dan 25% domba menunjukkan onset berahi antara 43,27 - 45,49 jam. Rata-rata onset berahi tercepat diperoleh pada kelompok domba yang diberi hormon prostaglandin F2 secara IM, yaitu 22,91 0,77 jam dari mulai di suntik prostaglandin F2 sampai munculnya awal berahi, sedangkan pemberian prostaglandin F2 secara IVS selama 6 hari menunjukkan onset berahi yang paling lama (44,57 1,09 jam).

Hasil penelitian ini relatif lebih cepat jika dibandingkan dengan domba Priangan yang diberi CIDR (Control Internal Drug Realising), onset berahi berkisar antara 24 58 jam dengan rataan 37.07 jam (Hastono et al., 2000) atau 36.33 jam pada domba St. Croix yang diberi flugeston asetat IVS (Hastono et al., 1997), dan 2 6 hari pada domba yang diberi Medroxy Progesterone Acetate (Sutama dan Dharsana, 1994). Sebagian besar onset berahi pada ternak antara 29 dan 48 jam (Trounson et al., 1976) atau pada rata-rata 44 jam (Acritopoulou et al., 1977). Perbedaan onset berahi ini dapat diakibatkan oleh perbedaan preparat hormon dan dosis 5-7,5 mg/ml yang diberikan, di samping faktor pengamatan, kondisi ternak, dan pakan yang diberikan (Toelihere, 2003). Teknik pemberian prostaglandin F2 berpengaruh sangat nyata (P<0,01) terhadap onset berahi pada domba. Pemberian prostaglandin F2 secara IM tidak berbeda nyata (P>0,05) dengan secara IVS yang diberikan selama 2 hari dalam vagina, namun keduanya berbeda sangat nyata (P<0,01) dengan secara IVS yang diberikan selama 4 dan 6 hari (Tabel 1). Hal tersebut menunjukkan bahwa pemberian prostaglandin F2 secara IVS selama dua hari cukup efektif dalam memacu munculnya awal tanda-tanda berahi. Perbedaan onset berahi diduga akibat adanya perbedaan kondisi fungsional CL, sesuai dengan pendapat Partodihardjo (1992), bahwa prostaglandin F2 sangat efektif untuk meregresikan CL yang sedang berfungsi, tetapi kurang efektif terhadap CL yang sedang tumbuh. Hal yang sama dinyatakan Hansel dan Schechter (1972), serta Rowson et al. (1972), bahwa sebelum stadium ini, CL yang sedang berkembang pada domba tampaknya tidak peka terhadap pengaruh luteolisis prostaglandin F2. Selanjutnya dinyatakan, bahwa penggunaan prostaglandin F2 pada ternak berahi sampai hari kelima setelah berahi, CL masih dalam keadaan tumbuh. Fase luteal dengan kondisi CL fungsional, pemberian prostaglandin F2 sangat efektif dalam meregresi CL sebagai sumber penghasil progesteron. Akibat adanya regresi CL, maka tidak ada lagi suplai progesteron. Tidak adanya progesteron, maka FSH dan LH disekresikan oleh kelenjar hipofise dan folikel sebagai sumber estrogen akan berkembang sehingga terjadi berahi. Sekresi korpora lutea dapat ditimbulkan dengan injeksi tunggal prostaglandin F2 IM (Douglas dan Ginther, 1973; Hawk, 1973) atau dengan injeksi analog prostaglandin F2 yang cocok (Hearnshaw et al., 1974) ketika korpora lutea

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paling sedikit berumur tiga hari (yaitu pada hari keempat siklus berahi).

Lama Berahi
Rata-rata lama berahi domba ekor tipis yang diberi prostaglandin F2 secara IM dan IVS dengan durasi yang berbeda secara rinci disajikan pada Tabel 2.
Tabel 2. Rataan lama berahi (jam) pada domba Perlakuan P 1 (IM) P 2 (IVS 2 hari) P 3 (IVS 4 hari) P 4 (IVS 6 hari)
abcd

Rata-rata 26,36 0,75 a 48,36 1,24 b 94,65 2,92 c 46,56 2,19 d

Superskrip yang berbeda pada kolom yang sama menunjukan ada perbedaan pada P<0,01

Berdasarkan Tabel 2 diketahui bahwa lama berahi pada domba ekor tipis berkisar antara 25,30 148,24 jam dengan rata-rata 78,98 jam sejak awal munculnya tanda-tanda berahi sampai berahi berakhir. Berahi paling pendek terjadi pada kelompok domba yang diberi hormon prostaglandin F2 secara IM, yaitu 26,36 0,75 jam. Pemberian prostaglandin F2 secara IVS selama 6 hari menunjukkan lama berahi yang paling panjang yaitu 146,56 2,19 jam. Lama berahi hasil penelitian ini relatif lebih panjang jika dibandingkan dengan domba St. Croix yang diberi flugeston asetat IVS 38.58 jam (Hastono et al., 1997); pada domba Priangan 42.87 jam (Tambayong, 1993); pada siklus pertama domba dewasa 31,5 jam (Sutama, 1987); dan pada domba jawa ekor tipis 33,1 jam (Bradford et al., 1986). Adanya variasi lama berahi tersebut kemungkinan disebabkan oleh beberapa faktor, diantaranya variasi dalam pengamatan berahi, umur ternak, kesehatan dan bobot badan ternak (Toelihere, 2003). Perbedaan teknik pemberian prostaglandin F2 berpengaruh sangat nyata (P<0,01) terhadap lama berahi pada domba. Pemberian prostaglandin F2 secara IM berbeda sangat nyata (P<0,01) dengan teknik pemberian secara IVS baik yang diberikan selama 2, 4, maupun 6 hari. Lama berahi antar perlakuan secara IVS berbeda sangat nyata (P<0,01) (Tabel 2). Hal tersebut menunjukan bahwa lama berahi meningkat sejalan dengan lama pemberian prostaglandin F2 secara IVS. Pengamatan berahi berdasarkan lendir vagina bisa kurang tepat karena dengan pencabutan spons 4 dan 6 hari domba masih mengeluarkan lendir jadi seolah-olah domba masih dalam keadaan berahi yang sebetulnya hal tersebut

hanya lendir vagina akibat adanya spons di dalamnya. Perbedaan lama berahi ini diduga erat kaitannya dengan sifat farmakologis dan biokimia prostaglandin F2 yang dapat mengiritasi otot polos vagina, sesuai dengan pendapat Harper et al. (1979), bahwa prostaglandin F2 sangat efektif dalam mengaktivasi otot polos, disamping mempunyai efek inflamatori, vasodilatasi pembuluh darah, dan mengelusidasi cairan (McDonald, 1980; Felig et al., 1987). Adanya spons yang mengandung prostaglandin F2 di dalam vagina meningkatkan sekresi cairan (lendir) dan terjadi oedema sehingga tanda-tanda berahi seolah tampak terus, yang pada akhirnya akan memperpanjang waktu pengamatan lama berahi. Dengan demikian indikasi lama berahi yang diberi prostaglandin F2 4 dan 6 hari tidak dijadikan indikator yang baik untuk lama berahi.

Persentase Kebuntingan
Sebelum domba jantan digunakan sebagai pemacek, terlebih dahulu dilakukan pemeriksaan sperma secara makroskopis dan mikroskopis. Pemeriksaan dimaksudkan untuk mengetahui apakah domba jantan dapat digunakan sebagai pemacek yang baik untuk menghasilkan keturunan dengan kualitas yang optimal. Hasil dari pemeriksaan sperma secara makroskopis adalah warna sperma putih susu/krem; volume 1 ml; konsistensi pekat (kental); derajat keasaman (pH) 6,2 dan mempunyai bau yang spesifik. Hasil dari pemeriksaan mikroskopis kualitas sperma adalah gerak massa minimal ++ artinya gerak massa sperma berupa gelombang awan tipis, motilitas sperma 80% (spermatozoa progresif, gerakan sangat cepat); konsentrasi sperma minimal 3.0 billion/ml semen. Tabel 3 menunjukkan bahwa persentase kebuntingan berdasarkan NR 30 hari pada domba mencapai 60%. Persentase kebuntingan tertinggi diperoleh pada kelompok domba yang diberi hormon prostaglandin F2 secara intramuskuler dan secara IVS selama 2 hari, yaitu masing-masing 100%. Kelompok domba yang diberi prostaglandin F2 secara IVS selama 4 dan 6 hari masing-masing hanya mencapai 20%. Hal tersebut karena saat kawin terlalu lambat sehingga kondisi ovum sudah mulai rusak, akibatnya kalaupun terjadi fertilisasi, maka kematian embrionalnya cukup tinggi atau karena spermatozoa belum mengalami proses kapasitasi secara sempurna sehingga kemampuan untuk membuahi ovum lebih rendah. Secara keseluruhan hasil temuan ini relatif lebih tinggi (60%) jika dibandingkan dengan persentase

Efek Pemberian Prostaglandin F2 (Saoeni)

133

kebuntingan pada kambing yang menggunakan hormon progestagen yang mencapai 27,33% dengan kisaran 0 83% (Sianturi et al., 1997).
Tabel 3. Jumlah ternak bunting dan tidak bunting dari masing-masing perlakuan Perlakuan Ternak Bunting ekor % 5 10 5 10 1 20 1 20 12 60 Ternak tidak bunting ekor % 0 0 0 0 4 4 8

Ditjenak, 1983. Buku Harian PPS PPL Sub Sektor Peternakan. Direktorat Bina Produksi Peternakan. Direktorat Jenderal Peternakan. Deptan. Jakarta. Evans, G., and W.M.C. Maxwell, 1987. Salomons Artificial insemination of Sheep and Goats. Butter Worths, England. Felig, P., J.D. Baxter, A.E. Broadus and L.A. Frehman, 1987. Endocrinology and Metabolism. 2 nd Edition. McGraw-Hill Book Company. New York. Hlm. 1768 -1779. Hafez, E.S.E., 1987. Folliculogenesis, Egg Maturation and Ovulation Rate. In: Reproduction in Farm Animals. Lea & Febiger, Philadelphia. Hansel, W. and R.J Schechter, 1972. Biotechnical Procedures For Control of the Estrous cycles of Domestic Animals. Proceeding. 7 th International Congres Animal. Reproduction and Artificial Insemination, Munich Vol. 1, Pp.78-96. Hastono, I. Inounu dan N. Hidayati, 1997. Penyerentakan Berahi pada Domba St. Croix, Prosiding Seminar Nasional Peternakan dan Veteriner. Jilid II. Pusat Penelitian dan Pengembangan Peternakan. Bogor. Hlm. 457-461. Hastono, I. Inounu, A. Saleh dan N. Hidayati, 2000 Penyerempakan Berahi dengan menggunakan CIDR pada Domba Rakyat di Kecamatan Nagrag. Prosiding Seminar Nasional Peternakan dan Veteriner. Pusat Penelitian dan Pengembangan Peternakan. Bogor. Hlm. 143-148. Harper, H.A., V.W. Rodwell and P.A. Mayes, 1979. Review of Physiological Chemistry. 17 th Lange Medical Publications. Los Altos, California. Pp. 113. Hawk, H.W., 1973. Uterine Motility and Sperm Transport in the Estrous Ewe After Prostaglandin Induced Regression of Corpora Lutea. Journal Animal Science 37:1380-1385. Hearnshaw, H., B.J. Restall, C.D. Nancarrow and P.E. Mattner, 1974. Synchronisation of Oestrous in Cattle, Sheep and Goats using Prostaglandin Analogue. Proceeding Australian Society Animal Production 10: 242-245. Hunter, R.H.F., 1980. Physiology and Technology of Reproduction in Female Domestic Animals. Academic Press, San Francisco. McDonald, L.E., 1975. Veterinery Endocrinology and Reproduction. 2 nd. Edition. Lea and Febiger. Philladelphia. McDonald, L.E., 1980. Veterinary Endocrinology and Reproduction. Lea and Febiger Philadelphia. Hlm. 304-329.

P 1 (IM) P 2 (IVS 2 hari) P 3 (IVS 4 hari) P 4 (IVS 6 hari)

Penggunaan prostaglandin F2 untuk gertak berahi secara intramuskuler maupun IVS selama 2 hari tidak menunjukkan perbedaan kebuntingan pada domba percobaan. Hal ini sejalan dengan hasil penelitian Sumaryadi dan Manalu (1996), bahwa penggunaan prostaglandin F2 untuk penyerentakan berahi pada domba tidak mengganggu perkembangan folikel yang akan mengalami ovulasi.

Kesimpulan
Aplikasi gertak berahi dengan prostaglandin F2 secara intra vagina spons selama 2 hari paling efektif dan sama dengan secara intra muskuler untuk memperbaiki kinerja reproduksi ternak domba yang diindikasi dengan onset, lama berahi, dan keberhasilan kebuntingan mencapai 100 persen. Gertak berahi dengan prostaglandin F2 5 mg/ml/ekor yang diberikan secara intra muskuler maupun secara intra vagina spons selama 2 hari dapat diaplikasikan di lapangan.

Daftar Pustaka
Acritopoulou S., W. Haresign, J.P Foster and G.E. Lamming, 1977. Plasma Progesterone and L.H. Concentrations in ewes after injection of an analogue of prostaglandin F2. Journal Reproduction Fertility 49: 337-340. Bradford, G.E., J.F. Quirke, P. Sitorus, I. Inouner, B. Tiesnamurti, F.L. Bell, I.C. Fletcher and D.T. Torel, 1986. Reproduction in Javanese Sheep : Evidence for a Gene with Large Effect on Ovulation Rate and Litter size. Journal Animal Science 63: 418-431. Douglas, R.H., and O.J. Ginther, 1973. Luteolysis Following a single injection of Prostaglandin F2 in sheep. Journal Animal Science 37: 990-993.

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Partodihardjo, S., 1992. Ilmu Reproduksi Hewan. Penerbit Mutiara, Jakarta. Rowson, L.E.A., R. Tervit and A. Brand, 1972. The Use of Prostaglandins for Synchronisation of Oestrus in Cattle. Journal Reproduction Fertility 29:145 (Abstract). Sianturi, R.S.G., U. Adiati, Hastono, I.G.M. Budiarsana dan I.K. Sutama. 1997. Sinkronisasi Berahi Secara Hormonal pada Kambing Etawah. Prosiding Seminar Nasional Peternakan dan Veteriner. Pusat Penelitian dan Pengembangan Peternakan. Bogor. Hlm. 379 384. Sumaryadi, M.Y. dan W. Manalu, 1996. Hubungan antara jumlah folikel yang mengalami ovulasi terhadap keberhasilan kebuntingan domba pada berahi pertama setelah penyuntikan PGF2. Media Veteriner III (1): 25 33. Sumaryadi, M.Y., 2003. Perkembangan Bioteknologi Reproduksi Pada Ternak. Program Studi Sumber Daya Ternak. Program Pascasarjana. UNSOED. Sutama, I.K., 1987. Pubertal Development and Early Reproductive Performance of Javanese Thin-Tail

(JTT) Sheep. [Disertation] Universitas of New England. Australia. Sutama, I.K. dan R. Dharsana, 1994. Sinkronisasi birahi dan super ovulasi pada domba. Proceeding Seminar Sains dan Teknologi Peternakan. Hlm. 463-467. Tomaszewska M.W., I.K.Sutama, I.G. Putu dan T.D. Chaniago, 1991. Reproduksi Tingkah Laku dan Produksi Ternak di Indonesia. Gramedia Pustaka Utama. Jakarta. Tambayong, 1993. Pengaruh penggunaan gonadotrophin (PMSG+HCG) terhadap penampilan reproduksi domba periangan betina pada tingkat prolifikasi dan kondisi tubuh yang berbeda. [Tesis] Program Pascasarjana Universitas Padjadjaran, Bandung. Trounson, A.O., S.M. Willadsen and R.M. Moor, 1976. Effect of Prostaglandin analogue Cloprostenol on oestrus, ovulation and embryonic viability in sheep. Journal Agriculture Science 86: 609-611. Toelihere, M.R., 2003. Fisiologi Reproduksi pada Ternak. Penerbit PT. Angkasa, Bandung.

ANIMAL PRODUCTION, September 2007, hlm. 135 144 ISSN 1411 2027 Terakreditasi No.56/DIKTI/Kep/2005

Vol. 9 No.3

Pengaruh Pemberian Tepung Ikan Lokal dan Impor terhadap Pertambahan Bobot Badan, Tingkah Laku Seksual, dan Produksi Semen Kambing Kacang
(The Effect of Feeding Local and Imported Fish Meal on Daily Weight Gain, Sexual Performance, and Semen Production of Kacang Buck)
Marjuki Addulah1* , Kusmartono1 , Suyadi1 , Soebarinoto 1 dan Mohammad Winugroho2
1 2

Fakultas Peternakan, Universitas Brawijaya, Malang Balai Penelitian Ternak, Ciawi, Bogor

Formatted: Font: Not Italic, Indonesian, Expanded by 0,3 pt Formatted: Font: Not Italic, Indonesian, Expanded by 0,3 pt Formatted: Font: Not Italic, Indonesian, Expanded by 0,3 pt Formatted: Font: Not Italic, Indonesian, Expanded by 0,3 pt Formatted: Font: 11 pt, Not Bold, Not Italic, Indonesian, Expanded by 0,3 pt Deleted:

ABSTRACT: This research was aimed to compare quality of local fish meal and imported ones, mainly in term of their effects on daily weight gain, feed efficiency, sexual performance, and semen production of Kacang goat bucks. Fifteen bucks were allotted to Randomized Block Design with three treatments and 5 replications. Treatment A was concentrate containing 14.10% soy bean meal and 0.90% urea, treatment B and C were concentrate containing 15% of local and imported fish meal, respectively. Each buck was put in individual cage, fed on elephant grass ad libitum and the concentrate of 1.50% body weight. Variables measured were daily weight gain, feed efficiency, sexual performance, and semen production. Semen was collected twice a week for 8 weeks. The results showed that feeding concentrate either containing local fish meal (treatment B) or imported fish meal (treatment C) gave no significantly different effect on daily weight gain, feed efficiency, sexual performance, and semen production. However both treatments gave better effect on the variables than those feeding concentrate containing soy bean meal and urea (treatment A). Based on these results, it could be concluded that quality of local fish meal was not significantly different from imported ones. Thus, it can be used to argue the perception of fish meal consumers that quality of local fish meal is lower than imported ones. Key Words: Fish meal, local, goat

Pendahuluan
Aspek reproduksi merupakan salah satu kunci dalam suatu usaha peternakan. Berbagai faktor dapat mempengaruhi penampilan reproduksi ternak, antara lain kondisi fisiologis tubuh ternak, penyakit, pengamatan estrus, dan perkawinan serta faktor pakan. Pakan mempunyai peran sangat penting untuk proses reproduksi ternak. Robinson et al. (2005) menyatakan bahwa pakan berpengaruh secara langsung terhadap reproduksi melalui pasokan beberapa zat makanan yang dibutuhkan untuk proses perkembangan oosit dan spermatozoa, proses ovulasi, fertilisasi, daya tahan dan perkembangan embrio hingga lahir, dan secara tidak langsung melalui sintesis hormon reproduksi yang sangat penting dalam mengatur proses reproduksi.

* Korespondensi Penulis.: Marjuki 4663@yahoo.com

Beberapa zat makanan tertentu mempunyai peran penting dalam reproduksi ternak. Zat makanan tersebut adalah protein, lemak, mineral terutama Zn, dan vitamin terutama vitamin E sebagai antioksidan (Louis et al., 1994a; Louis et al., 1994b; Estienne dan Harper, 2004). Tepung ikan merupakan bahan pakan yang sangat baik sebagai sumber protein, lemak maupun mineral. Tepung ikan mengandung protein cukup tinggi yang tahan terhadap degradasi dalam rumen, dan mengandung lemak sekitar 10% yang sebagian besar berupa asam lemak tak jenuh yang sangat penting untuk sistem hormon reproduksi (Pike et al., 1994; Burke et al., 1997; FIN, 1999). Kualitas tepung ikan juga sangat bervariasi tergantung pada beberapa faktor, terutama kualitas bahan baku dan proses pembuatannya (Hussein dan Jordan, 1991; Pike et al., 1994; Aryawansa, 2000). Tepung ikan yang digunakan sebagai pakan ternak di Indonesia hampir 90% diimpor terutama dari Peru dan Chili, dan hanya 10% berasal dari produksi dalam negeri atau lokal. Konsumen lebih suka menggunakan tepung ikan impor dibanding lokal dengan alasan bahwa kualitas tepung ikan impor lebih tinggi dibanding tepung ikan lokal.

Deleted: sistem

Deleted: sperma Deleted: tozoa Deleted: ,

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Alasan tersebut tidak tepat, karena Marjuki (2007) melaporkan bahwa berdasarkan hasil uji laboratorium yang meliputi kadar bahan kering (BK), bahan organik (BO), protein kasar (PK), dan lemak; solubilitas, degradabilitas, dan daya cerna protein, komposisi asam amino dan asam lemak, kualitas tepung ikan lokal terutama tepung ikan yang diproduksi secara mekanik tidak berbeda dengan kualitas tepung ikan impor. Perbandingan kandungan PK antara tepung ikan lokal mekanik dan tepung ikan impor masing-masing adalah 59,10 dan 65,40% BK, lemak 10,90 dan 7,80% BK, total asam amino (tidak termasuk sistin, hidroksiprolin, prolin dan triptophan) 91,40 dan 91,30% PK, daya larut protein dalam air 22,90 dan 22,40%, proporsi protein potensial terdegradasi dalam rumen (in vitro) 35,20 dan 36,10%, kecernaan protein pascarumen (in vitro) 77,20 dan 81,20%. Penelitian ini bertujuan untuk menguji lebih jauh perbandingan kualitas tepung ikan lokal mekanik dan impor melalui uji biologis pengaruhnya terhadap pertambahan bobot badan, tingkah laku seksual, dan produksi semen kambing.

kacang jantan umur 12 - 18 bulan dengan rata-rata bobot badan awal 27,29 + 4,06 kg. Kambing tersebut ditempatkan dalam kandang individu yang dilengkapi tempat pakan dan minum. Pakan yang digunakan berupa rumput gajah dan tiga macam konsentrat sebagai pakan perlakuan, yaitu konsentrat perlakuan A mengandung 14,10% bungkil kedele dan 0,90% urea sebagai perlakuan kontrol, dan konsentrat B dan C berturut-turut mengandung 15% tepung ikan lokal mekanik dan tepung ikan impor. Komposisi bahan dalam masing-masing konsentrat disajikan pada Tabel 1 dan hasil analisis kandungan BK, BO, PK, dan lemak masing-masing konsentrat disajikan pada Tabel 2.

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Perlakuan dan Koleksi Data


Percobaan menggunakan Rancangan Acak Kelompok dengan 3 perlakuan yaitu perlakuan konsentrat A, B dan C (Tabel 1) dan diulang 5 kali menggunakan 5 kelompok kambing jantan yang dikelompokkan berdasarkan bobot badan awal. Penelitian ini dilaksanakan dalam dua tahap yaitu tahap adaptasi selama 3 minggu dan tahap perlakuan 8 minggu. Tahap adaptasi dilaksanakan untuk mengkondisikan kambing percobaan dengan kondisi penelitian terutama pemberian konsentrat. Selama tahap adaptasi semua kambing diberi pakan sama yaitu berupa rumput gajah ad libitum dan konsentrat perlakuan A sebanyak 1,50% bobot badan dalam bentuk BK.

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Metode Penelitian
Ternak dan Pakan
Penelitian menggunakan 15 ekor kambing

Tabel 1. Komposisi bahan pakan yang digunakan dalam masing-masing perlakuan (% total bobot) Bahan Bekatul (Oryiza sativa) Pollard (Triticum spp.) Bungkil Kelapa (Cocos nucifera) Bungkil Biji Kapok (Ceiba petandra) Jagung (Zea mays) Mineral Bungkil Kedele (Glycine max) Urea Tepung ikan lokal (Sardinella longisep) Tepung ikan impor (Engraulis ringens) Perlakuan A 40,00 18,00 12,00 5,00 8,00 2,00 14,10 0,90 0,00 0,00 B 40,00 18,00 12,00 5,00 8,00 2,00 0,00 0,00 15,00 0,00 C 40,00 18,00 12,00 5,00 8,00 2,00 0,00 0,00 0,00 15,00

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Perlakuan A mengandung 14,10% bungkil kedele dan 0,90% urea sebagai perlakuan kontrol; Perlakuan B dan C berturut-turut mengandung 15% tepung ikan lokal mekanik dan tepung ikan impor.

Pengaruh Pemberian Tepung Ikan (Marjuki et al.)

139

Tabel 2. Hasil analisis kandungan zat makanan rumput gajah dan konsentrat masing-masing perlakuan Pakan Rumput gajah pemberian Konsentrat : - Perlakuan A - Perlakuan B - Perlakuan C 87,20 87,60 87,40 88,70 86,30 86,40 20,10 19,70 19,80 7,00 7,80 7,70 BK (%) 20,40 BO (% BK) 83,70 PK (% BK) 9,50 LK (% BK) 2,50

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BK: bahan kering, BO: bahan organik, PK: protein kasar. Perlakuan A mengandung 14,10% bungkil kedele dan 0,90% urea sebagai perlakuan kontrol; perlakuan B dan C berturut-turut mengandung 15% tepung ikan lokal mekanik dan tepung ikan impor.

Tahap perlakuan dilaksanakan untuk mengetahui pengaruh pemberian konsentrat A, B, dan C terhadap pertambahan bobot badan, tingkah laku seksual, dan kualitas semen. Masing-masing kambing dalam satu kelompok diberi pakan perlakuan berbeda yaitu konsentrat A, B, dan C sebanyak 1,50% bobot badan ditambah rumput gajah dan air minum secara ad libitum. Selama tahap perlakuan dilaksanakan pengukuran konsumsi pakan harian (BK, BO, PK dan LK), pengamatan tingkah laku seksual, koleksi atau pengamatan produksi semen dan pengamatan profil darah (kadar kolesterol, urea, dan hormon testosteron). Koleksi semen dilaksanakan dua kali seminggu menggunakan vagina buatan dengan kambing betina sebagai pemancing. Semen yang dikoleksi dari masing-masing kambing segera diuji secara makrokopis untuk variabel volume, warna (skor : 0 = bening, 1 = putih, 2 = putih susu, 3 = krem), pH, dan konsistensi semen (skor : 1 = encer, 2 = sedang, 3 = kental), dan secara mikrokopis untuk variabel motilitas massa (skor : 0 = Gerakan sangat lambat, 1 = Gerakan lambat, 2 = gerakan cepat dan gelombang tipis, 3 = Gerakan sangat cepat dan gelombang tebal), motilitas individu, konsentrasi spermatozoa (jumlah spermatozoa per ml semen), persentase spermatozoa hidup (viabilitas) dan jumlah spermatozoa per ejakulasi. Tingkah laku seksual diamati sesuai dengan prosedur Herwijanti (2004) dan dilakukan pada saat pelaksanaan koleksi semen, meliputi : a. Libido yaitu waktu (detik) yang diperlukan oleh pejantan mulai dari saat didekatkan dengan betina pemancing sampai melakukan false mounting (menaiki betina pemancing). b. Jumlah false mounting yaitu berapa kali kambing pejantan melakukan false mounting sampai terjadi ejakulasi.

c. Daya jepit yaitu kemampuan pejantan untuk menekankan kedua kaki depannya pada otot semi membraneous kambing betina pemancing saat terjadinya ejakulasi (nilai +++ = kuat yaitu jika kaki depan pejantan menjepit tepat pada latero lumbal kambing betina pemancing; ++ = sedang yaitu jika jepitan kaki depan pejantan kurang mantap, + = lemah yaitu jika kaki pejantan mlorot atau tidak menjepit). d. Daya dorong yaitu kemampuan pejantan untuk mendorongkan tubuhnya pada kambing betina pemancing pada saat ejakulasi (nilai +++ = kuat, yaitu jika kaki belakang pejantan ikut melompat; ++ = sedang yaitu jika terjadi perubahan posisi kaki belakang pejantan tetapi tidak melompat; + = lemah yaitu jika posisi kaki belakang pejantan tidak berubah dan tidak melompat). e. Kualitas ereksi yaitu penampakan organ kelamin pejantan pada saat ereksi (nilai +++ = baik yaitu jika warna penis merah disertai atau tanpa disertai cairan seminal plasma; ++ = sedang yaitu jika warna penis merah muda sampai merah muda pucat; + = jelek yaitu jika penis tidak keluar dari preputium). f. Lama ejakulasi yaitu waktu (detik) yang diukur mulai dari saat pejantan didekatkan pemancing sampai terjadi ejakulasi. Konsentrasi hormon testosteron, kolesterol, dan urea darah dianalisa pada sampel darah yang diambil dari vena jugularis masing-masing kambing sebanyak 3 ml per ekor pada awal minggu pertama, keempat dan kedelapan tahap perlakuan. Sampel darah diambil sehari sebelum waktu koleksi semen mulai jam 10.00 sampai 11.00 dengan interval pengambilan setiap 15 menit. Sampel darah yang telah diambil segera ditransfer ke tabung reaksi kapasitas 10 ml yang di dalamnya telah diisi zat antikoagulan larutan EDTA 10% sebanyak 0,5 ml, kemudian dikocok 3 sampai 4 kali secara pelan-

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pelan. Sampel darah masing-masing kambing segera disentrifuge pada kecepatan 3000 rpm selama 15 menit, kemudian bagian supernatan dan ditransfer ke sebuah vial kapasitas 0,5 ml menggunakan spuit. Sampel supernatan tersebut kemudian disimpan dalam freezer suhu 20o C hingga semua sampel selama penelitian terkumpul. Pada akhir penelitian semua sampel dikomposit berdasarkan individu kambing dan periode pengambilan, kemudian diambil sub sampel sebanyak 1 ml untuk dianalisis konsentrasi hormon testosteron, kadar kolesterol dan urea. Analisis konsentrasi hormon testosteron dilaksanakan di Laboratorium Endokrinologi Rumah Sakit Umum Daerah Dr. Soetomo Surabaya, sedang analisis kadar kolesterol dan urea darah dilaksanakan di Laboratorium Klinik Sigma Surabaya.

Analisis Data
Respon data kuantitatif terhadap perlakuan dianalisis ragam dengan menggunakan program komputer SPSS 10.0 for Windows (SPSS, Inc. 1999), sedangkan data kualitatif dianalisis secara deskriptif.

Hasil dan Pembahasan


Konsumsi Pakan, Pertambahan Bobot Badan dan Konversi Pakan. Konsentrat yang tidak mengandung tepung ikan (perlakuan A) menunjukkan palatabilitas lebih tinggi dibanding konsentrat B dan C yang masing-masing mengandung tepung ikan lokal dan tepung ikan impor sebanyak 15% (Tabel 3). Hal tersebut ditunjukkan oleh persentase jumlah konsentrat tersisa yang secara nyata lebih kecil (P<0,05) pada konsentrat A (2,27%) dibanding pada konsentrat B (6,89%) dan C (15,87%), dan data persentase konsumsi BK konsentrat terhadap konsumsi BK total yang secara nyata (P<0,05) lebih tinggi pada konsentrat perlakuan A (49,31%) diikuti konsentrat B (46,63%) dan C (44,52%). Perbedaan palatabilitas tersebut disebabkan oleh aroma tepung ikan yang dapat menyebabkan rendahnya palatabilitas pakan terutama pada ternak ruminansia, sebagaimana dinyatakan Schroeder (1999) dan Stallings (2003) bahwa tepung ikan merupakan bahan pakan yang kaya protein tetapi palatabilitasnya rendah, terutama bagi ternak ruminansia. Di antara konsentrat perlakuan yang menggunakan tepung ikan, konsentrat B menunjukkan palatabilitas lebih tinggi (P<0,05) daripada konsentrat C. Tepung ikan lokal memiliki rasa lebih asin yang dapat meningkatkan palatabilitas pakan

karena kondisi bahan baku yang digunakan sudah tidak segar, sehingga lebih banyak menyerap rasa asin dari air laut terutama selama selang waktu saat ikan ditangkap hingga disortir dan diproses menjadi tepung ikan dibanding tepung ikan impor yang menggunakan ikan segar sebagaimana dinyatakan oleh Hussein dan Jordan (1991). Perlakuan A menunjukkan konsumsi total BK, BO, PK dan LK paling tinggi, tetapi menunjukkan pertambahan bobot badan paling rendah dan angka konversi pakan paling tinggi (P<0,05) dibanding perlakuan B dan C. Hal ini menunjukkan bahwa efisiensi pemanfaatan zat makanan terkonsumsi oleh ternak pada konsentrat perlakuan B dan C lebih tinggi dibanding konsentrat perlakuan A. Hussein dan Jordan (1991) yang merangkum beberapa hasil penelitian terdahulu; Grigsby et al. (1999); Veira et al. (1988); Zinn dan Owens (1993); Rocha et al. (1995) melaporkan bahwa subtitusi bungkil kedelai, atau bungkil kedelai yang dicampur urea dengan tepung ikan dapat menghasilkan pertambahan bobot badan lebih tinggi dan konversi pakan lebih baik. Lebih lanjut Rocha et al. (1995) menyatakan bahwa tepung ikan merupakan bahan pakan sumber ruminally undegradable protein (RUP) dan kaya akan lisin dan methionin yang merupakan dua asam amino pembatas pada ternak ruminansia. Tepung ikan umumnya mengandung RUP lebih dari 70% sedangkan bungkil kedele mengandung RUP kurang dari 45 % dan tepung ikan dapat memasok lisin dan methionin masing-masing dua dan empat kali lipat lebih besar dibanding bungkil kedele (Blauwiekel et al., 1992). Penampilan Tingkah Laku Seksual Pemberian pakan perlakuan berpengaruh nyata (P<0,05) terhadap waktu libido, waktu ejakulasi, dan jumlah mounting (Tabel 4). Perlakuan B menunjukkan waktu libido dan waktu ejakulasi lebih pendek dan jumlah mounting lebih sedikit dibanding perlakuan A dan C yang keduanya tidak berbeda nyata antara satu dengan lain. Lebih dari 50% kambing pada masing-masing perlakuan menunjukkan daya jepit yang kuat, namun pemberian konsentrat yang mengandung tepung ikan menunjukkan persentase kambing dengan daya jepit kuat sebesar 75,30% pada perlakuan B dan 60,00% pada perlakuan C, lebih besar dibanding pada perlakuan A yaitu sebesar 56,50%. Pola yang sama juga terjadi pada daya dorong maupun kualitas ereksi. Hal ini menunjukkan bahwa pemberian tepung ikan memberikan pengaruh lebih baik dalam

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Deleted: , Deleted: dan Deleted: , Deleted: ), Deleted: Deleted: serta Deleted: Deleted: Hal ini karena tepung ikan mempunyai kualitas lebih baik dibanding bungkil kedele maupun bungkil kedele plus urea, terutama jika dilihat dari degradabilitas proteinnya yang rendah dalam rumen dan komposisi asam amino dan asam lemaknya yang lebih baik (Hussein dan Jordan, 1991; FIN, 2000) Deleted: juga Deleted: Deleted: Formatted: Font: 11 pt, Italic, Indonesian, Character scale: 98%, Expanded by 0,3 pt Formatted: Font: 11 pt, Italic, Indonesian, Character scale: 98%, Expanded by 0,3 pt Deleted: pada Deleted: yang Deleted: Pemberian pakan perlakuan juga berpengaruh nyata (P<0,05) terhadap jumlah mounting, di mana walaupun tidak berbeda nyata, jumlah mounting pada perlakuan B lebih sedikit dibanding perlakuan C yang keduanya secara nyata lebih sedikit dibanding pada perlakuan A. Formatted: Font: 11 pt, Italic, Indonesian, Character scale: 98%, Expanded by 0,3 pt Formatted: Font: 11 pt, Italic, Indonesian, Character scale: 98%, Expanded by 0,3 pt Deleted: oleh Deleted: Anonymous (2001a) Deleted: Sedangkan di

Pengaruh Pemberian Tepung Ikan (Marjuki et al.)

139

Tabel 3. Rata-rata konsumsi, pertambahan bobot badan, dan konversi pakan Parameter A Konsumsi : - BK total (g/ekor/hari) - BK total (% BB) - BK total (g/kg BB 0.75/hari) - BO total (g/kg BB 0.75/hari) - PK total (g/kg BB
0.75

Perlakuan B 952,10 160,20 b 3,12 0,43 a 73,18 9,44a 62,28 7,94 b 10,64 1,32
b

C 910,70 123,80 a 3,08 0,39 a 71,71 8,36 a 60,90 7,18 a 10,39 1,28 a 3,54 0,51 a 44,52 8,52a 15,87 19,00c 77,49 76,30
a b

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968,80 91,40 b 3,29 0,38 b 76,57 7,92 b 66,06 6,67 c 11,99 0,82
c

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/hari)

- LK total (g/kg BB 0.75 /hari) - BK konsentrat (% dari BK total) Persentase konsentrat tersisa (%) PBB (g/ekor/hari) Konversi pakan (kg pakan/kg PBB)

3,71 0,27 b 49,31 5,10 c 2,27 6,97a 70,30 47,10


a

3,69 0,50 b 46,63 7,62 b 6,89 15,30 b 101,63 67,80 9,37 2,36 a

13,78 1,94 c

11,75 1,62b

a,b Superskrip yang berbeda pada baris yang sama menunjukkan ada perbedaan pada P<0,05. BK : bahan kering, BO: bahan organik, PK: protein kasar, LK: lemak kasar, BB: bobot badan, PBB: pertambahan bobot badan. Perlakuan A mengandung 14,10% bungkil kedele dan 0,90% urea sebagai perlakuan kontrol; perlakuan B dan C berturut-turut mengandung 15% tepung ikan lokal mekanik dan tepung ikan impor

Tabel 4. Data tingkah laku seksual kambing pada masing-masing perlakuan Parameter A (n = 85) Libido (detik) Ejakulasi (detik) Mounting (kali) Daya jepit : - Lemah - Sedang - Kuat Daya dorong : - Lemah - Sedang - Kuat Kualitas ereksi : - Jelek - Sedang - Baik
a,b

Formatted Formatted Formatted

Perlakuan B (n = 85) 7,77 6,31 a 17,38 9,63 a 3,21 1,41 a Frek. Persen 2 19 64 1 17 67 0 4 81 2,40 22,40 75,30 1,20 20,00 78,80 0 4,70 95,30 C (n = 85) 12,03 + 25,36 + 3,56 + Frek. 1 33 51 1 18 66 0 6 79 11,18 b 14,00 b 1,64 ab Persen 1,20 38,80 60,00 1,20 21,20 77,60 0 7,10 92,90 11,05 9,91 b 23,25 13,53 b 3,67 1,66 b Frek. Persen 5 32 48 3 37 45 0 11 74 5,90 37,60 56,50 3,50 43,50 52,90 0 12,90 87,10

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Superskrip yang berbeda pada baris yang sama menunjukkan ada perbedaan pada P<0,05. Perlakuan A mengandung 14,10% bungkil kedele dan 0,90% urea sebagai perlakuan kontrol; perlakuan B dan C berturutturut mengandung 15% tepung ikan lokal mekanik dan tepung ikan impor

meningkatkan tingkah laku seksual (daya jepit, daya dorong maupun kualitas ereksi) kambing dibanding pemberian bungkil kedelai dan urea. Beberapa faktor dapat berpengaruh terhadap penampilan tingkah laku seksual seekor ternak, di

antaranya adalah pakan. Ternak yang mendapatkan pakan lebih baik umumnya menunjukkan tingkah laku seksual lebih baik (Louis et al., 1994a; Louis et al., 1994b).

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Produksi dan Kualitas Semen


Hasil pengamatan evaluasi semen dalam penelitian ini disajikan pada Tabel 5 yang menunjukkan bahwa secara umum pemberian tepung ikan dalam konsentrat sebagai pakan perlakuan berpengaruh nyata (P<0,05) terhadap kualitas semen. Volume semen merupakan salah satu kriteria penting dalam mengevaluasi kualitas semen dan penampilan reproduksi ternak jantan (Ax et al., 2000). Rata-rata volume semen per ejakulasi pada penelitian ini berkisar antara 0,72 sampai 0,86 ml. Hal ini sesuai dengan pendapat Wildeus (2000); Haenlein et al. (2006) bahwa volume semen per ejakulasi pada kambing berkisar antara 0,5 sampai 1,5 ml. Pemberian pakan supelemen tepung ikan dalam konsentrat berpengaruh nyata (P<0,05) terhadap volume semen. Perlakuan C memberikan volume semen paling banyak yaitu 0,85 ml per ejakulasi dibanding perlakuan B yang tidak berbeda nyata dengan perlakuan A, yaitu masing-masing 0,76 dan 0,73 ml per ejakulasi. Rata-rata derajat warna semen yang dikoleksi selama penelitian berkisar antara 2,25 sampai 2,47 atau berwarna putih-krem sampai krem. Hasil pengamatan warna tersebut sesuai dengan pendapat Oyeyemi et al. (2000) bahwa semen kambing berwarna krem atau putih susu. Warna dan derajat kekeruhan juga berpengaruh terhadap derajat kekentalan atau konsistensi semen. Semen yang bening umumnya bersifat encer, dan konsistensi semakin kental pekat sehubungan dengan meningkatnya derajat kekeruhan dan warna. Konsistensi semen dalam penelitian ini berkisar antara 2,60 sampai 2,69 atau agak kental sampai kental. Derajat keasaman atau pH merupakan salah satu sifat kimia semen yang dipengaruhi oleh proses pemanfaatan zat makanan cadangan dalam semen oleh spermatozoa dan atau mikroorganisma kontaminan dalam semen. Penurunan pH semen menunjukkan telah terjadinya penguraian atau pemanfaatan zat makanan khususnya fruktosa dalam semen, sehingga semakin rendah pH semen berarti semakin berkurang ketersediaan zat makanan dalam semen untuk spermatozoa. Kondisi tersebut juga akan mempengaruhi daya hidup spermatozoa dalam semen. Semen yang normal mempunyai pH berkisar antara 6,5 7,0 (Salisbury et al., 1978). Rata-rata pH semen hasil pengamatan penelitian ini berkisar antara 6,98 pada perlakuan B yang tidak berbeda nyata (P>0,05) dengan perlakuan A dan C yang keduanya menunjukkan pH semen 7,05.

Persentase spermatozoa hidup (viabilitas), motilitas massal dan motilitas individu progresif merupakan indikator yang sangat penting dalam evaluasi semen. Ketiga indikator tersebut sangat menentukan kemampuan spermatozoa untuk dapat bergerak dalam menemukan oosit untuk proses fertilisasi. Hasil pengataman dalam penelitian ini menunjukkan bahwa pemberian tepung ikan dalam konsentrat berpengaruh nyata terhadap viabilitas spermatozoa. Rata-rata viabilitas spermatozoa pada penelitian ini berkisar antara 64,95% pada perlakuan A yang secara nyata (P<0,05) lebih kecil dibanding perlakuan B dan C, yang masing-masing menunjukkan viabilitas spermatozoa 69,22 dan 71,32%. Hasil pengamatan tersebut sesuai dengan Hullet dan Shelton (1987) yang menyatakan bahwa semen kambing yang normal mempunyai viabilitas antara 60 sampai 80%. Pola yang sama juga terjadi pada hasil pengamatan motilitas massa dan individu spermatozoa. Rata-rata motilitas massa dan individu spermatozoa pada perlakuan A (2,26 dan 61,71%), lebih rendah daripada perlakuan B (2,47 dan 68,65%) dan perlakuan C ( 2,52 dan 71,71%). Hasil pengamatan motilitas massa spermatozoa tersebut sesuai dan bahkan pada perlakuan B dan C lebih tinggi dibanding pendapat Haenlein et al. (2006) yang menyatakan bahwa semen segar yang baik minimal harus menunjukkan motilitas spermatozoa antara 60 sampai 70%. Konsentrasi spermatozoa merupakan indikator yang menunjukkan jumlah spermatozoa per mililiter semen. Hal tersebut sangat penting untuk diketahui terutama jika semen harus diencerkan untuk program inseminasi buatan. Hasil penelitian ini menunjukkan bahwa pemberian tepung ikan dalam konsentrat berpengaruh nyata terhadap konsentrasi spermatozoa dalam semen. Rata-rata konsentrasi spermatozoa pada perlakuan A (247,89 + 84,89 x 10 7 spermatozoa per ml semen) secara nyata lebih tinggi (P<0,05) dibanding perlakuan B (214,66 + 73,11 x 10 7) dan perlakuan C (221,91 + 62,89 x 10 7). Hasil tersebut lebih rendah dibandingkan pendapat Wildeus (2000) sebesar 3 x 10 9 atau berkisar 1,5 5,0 x 109 spermatozoa per ml semen. Perlakuan pemberian tepung ikan dalam konsentrat tidak berpengaruh nyata (P>0,05) terhadap jumlah spermatozoa per ejakulasi, namun berpengaruh nyata (P<0,05) terhadap jumlah spermatozoa progresif per ejakulasi dengan nilai tertinggi pada perlakuan C 136,85 + 81,23x10 7 spermatozoa per ejakulasi, kemudian perlakuan A (119,06 + 82,79x10 7) dan perlakuan B (112,00 + 87,20x10 7). Kheradmand et al. (2006) menyatakan

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bahwa penampilan produksi dan reproduksi ternak dipengaruhi oleh empat faktor yaitu faktor genetik, kondisi lingkungan fisik, pakan atau nutrisi, dan manajemen. Lotthammer (1991) menyatakan bahwa kurang lebih 80% faktor yang mempengaruhi fertilitas ternak adalah faktor lingkungan, dari nilai tersebut 50% nya adalah karena faktor pakan terutama jika faktor penyakit dan fertilitas pasangannya diabaikan. Meskipun faktor penyakit sebenarnya juga dapat disebabkan atau diatasi dengan faktor pakan, sehingga faktor pakan sangat dominan dalam menentukan produktivitas, kesehatan dan fertilitas ternak (Robinson et al., 2005).

Konsentrasi Kolesterol, Urea dan Hormon Testosteron Darah


Hasil pengukuran konsentrasi hormon testosteron dalam penelitian ini berkisar antara 12,88 sampai 17,14 nMol/L (Tabel 6). Hasil tersebut sesuai dengan pendapat Delgadillo et al. (2002), Delgadillo et al. (2004) dan Kridli et al. (2006) yaitu berkisar antara 2,80 + 0,60 ng/ml sampai 13,00 + 3,60 ng/ml atau jika berat molekul hormon testosteron adalah 288,40 g (Diagnostic Automation, Inc., 2001), maka konsentrasi tersebut setara dengan 9,71+2,08 nmol/L sampai 45,07+12,48 nmol/L. Kolesterol disintesis terutama untuk pembentukan silomikron dalam proses penyerapan
Tabel 5. Kualitas semen pada masing-masing perlakuan Parameter

lemak, sehingga semakin besar jumlah konsumsi dan penyerapan lemak semakin banyak kolesterol yang dibutuhkan dan disintesis oleh tubuh (Grummer dan Carroll, 1991). Kadar kolesterol dalam darah (cholesterol pool) dipengaruhi oleh jumlah dan kualitas lemak yang dikonsumsi oleh seekor ternak terutama degradabilitasnya dalam rumen dan penyerapannya dari usus halus (intake) dan penggunaan kolesterol oleh tubuh termasuk untuk pembentukan hormon-hormon steroid (uptake). Kolesterol merupakan bakalan untuk pembentukan hormon testosteron pada hewan jantan atau progesteron pada hewan betina (Ogle dan Costoff, 2001), sehingga semakin tinggi konsentrasi hormon androgen (steroid) dalam darah memungkinkan berkurangnya konsentrasi kolesterol darah sebagaimana ditunjukkan oleh data pada Tabel 6. Hasil pengukuran kadar urea darah dalam penelitian ini berkisar antara 29,80 hingga 33,00 mg/dL. Kohn et al. (2005) melaporkan bahwa kadar urea darah pada kambing berkisar antara 25 hingga 38 mg/dL. Nilai hampir sama dilaporkan Rivas dan Serrato-Corona (2005) bahwa kadar urea darah pada kambing yang diberi pakan mengandung rumen degradable protein (RDP) tinggi sebesar 28,3 mg/dL dan yang diberi pakan mengandung RDP rendah sebesar 25,5 mg/dL.

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Perlakuan A (n = 85) B (n = 85)


a

C (n = 85)
a

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b

Volume (ml) Warna PH Konsistensi Viabilitas (%) Motilitas massa Motilitas individu (%) Konsentrasi (x 107 sel spermatozoa per ml semen) Total spermatozoa per ejakulasi (x 10 7 ) Total spermatozoa progresif per ejakulasi (x 10 )
a,b

0,72 0,27

0,76 0,29

0,86 0,25

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Putih-krem (skor : 2,45 0,73) 7,05 0,17 Kental (skor : 2,69 0,49) 64,95 15,86 a ++ (skor : 2,26 0,88) 65,71 18,80 a 247,89 84,89 b 178,48 106,32
7

Putih-krem (skor : 2,25 0,95) 6,98 0,26 Kental (skor : 2,60 0,58) 69,22 15,27 b ++ (skor : 2,47 0,70) 68,65 15,11b 214,66 73,11 a 163,14 103,37

Putih-krem (skor : 2,47 0,85) 7,05 0,21 Kental (skor : 2,67 0,50) 71,32 11,19b ++ (skor : 2,52 0,70) 71,71 14,75 b 221,91 62,89 a 190,83 93,85

117,28 82,79

ab

112,00 87,20

136,85 81,23 b
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superskrip yang berbeda pada baris yang sama menunjukkan ada perbedaan pada P<0,05. Perlakuan A mengandung 14,10% bungkil kedele dan 0,90% urea sebagai perlakuan kontrol; perlakuan B dan C berturut-turut mengandung 15% tepung ikan lokal mekanik dan tepung ikan impor

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Tabel 6. Rata-rata konsentrasi hormon testosteron, urea dan kolesterol pada masing-masing perlakuan dan periode koleksi sampel Variabel Sampel A Minggu 1 Testosteron (nMol/L) Minggu 4 Minggu 8 Minggu 1,4,8 Minggu 1 Urea darah (mg/dL) Minggu 4 Minggu 8 Minggu 1,4,8 Minggu 1 Kolesterol (mg/dL) Minggu 4 Minggu 8 Minggu 1,4,8 12,88 7,69 13,56 7,36 13,60 8,26 13,35 7,21 32,00 5,00 30,80 7,09 31,80 5,07 31,53 5,40 44,00 9,41 47,60 12,32 49,60 7,13 47,07 9,43 Perlakuan B 12,66 5,09 12,94 6,88 17,14 10,29 14,25 7,46 32,40 4,22 29,80 4,09 30,20 4,21 30,80 4,04 43,40 7,77 45,80 8,87 48,00 7,14 45,73 7,62 C 13,30 6,59 14,06 5,80 13,86 6,94 13,77 5,94 33,00 5,29 30,20 5,63 30,20 8,35 31,00 6,28 45,75 7,18 45,20 14,10 48,80 4,60 46,64 9,08

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Perlakuan A mengandung 14,10% bungkil kedele dan 0,90% urea sebagai perlakuan kontrol; perlakuan B dan C berturut-turut mengandung 15% tepung ikan lokal mekanik dan tepung ikan impor

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Pemberian pakan perlakuan maupun lama masa pemberiannya tidak berpengaruh nyata (P>0,05) terhadap konsentrasi testosteron, urea maupun kolesterol darah, tetapi konsentrasi hormon testosteron dan kolesterol menunjukkan kecenderungan meningkat, sebaliknya konsentrasi urea menunjukkan kecenderungan menurun sehubungan dengan lamanya pemberian pakan perlakuan. Hal tersebut menunjukkan adanya peningkatan efisiensi pemanfaatan pakan karena ternak semakin teradaptasi dengan kondisi pakan yang diberikan. Beberapa faktor berpengaruh terhadap produksi hormon testosteron, di antaranya adalah faktor musim terutama pada ternak yang mempunyai musim kawin pada musim tertentu dan faktor pakan sebagaimana yang dinyatakan oleh Volek et al. (1997), Walkden-Brown et al. (2006), Delgadillo et al. (2002) dan Delgadillo et al. (2004). Konsumsi protein yang melebihi kebutuhan terutama yang mudah mengalami degradasi dalam rumen atau konsumsi energi dari hasil konversi protein dengan hasil samping berupa amonia atau urea darah dilaporkan menyebabkan rendahnya konsentrasi hormon testosteron dalam darah, sebaliknya konsumsi energi terutama yang berasal dari lemak menyebabkan meningkatnya konsentrasi hormon testosteron dalam darah (Volek et al., 1997).

Kadar urea darah pada ternak ruminansia merupakan hasil dari proses pencernaan dan metabolisme protein atau senyawa nitrogen yang lain. Sebagian senyawa nitrogen terdegradasi di dalam rumen oleh mikroba menjadi amonia, yang kemudian sebagian amonia diserap oleh tubuh melalui dinding rumen dan oleh peredaran darah dibawa ke hati. Di dalam hati amonia semaksimal mungkin diubah menjadi urea yang kemudian dibawa oleh peredaran darah dan dikeluarkan melalui urine dan sebagian dikembalikan ke rumen melalui saliva dan dinding rumen. Perlakuan A menunjukkan konsentrasi kolesterol dan urea darah lebih tinggi tetapi konsentrasi hormon testosteron lebih rendah dibanding perlakuan B dan C. Hal ini karena sumber protein dalam pakan perlakuan A yaitu bungkil kedele dan urea mempunyai degradabilitas lebih tinggi dibanding sumber protein dari tepung ikan pada pakan perlakuan B dan C (Blauwiekel and Harrison., 1992), sehingga menyebabkan konsentrasi urea darah yang lebih tinggi pada perlakuan A dibanding perlakuan B dan C. Menurut Volek et al. (1997) bahwa konsumsi protein yang terlalu tinggi dalam pakan dan degradabilitas dalam rumen yang tinggi serta konversi protein menjadi energi dengan hasil samping berupa amonia atau urea darah dapat menyebabkan rendahnya konsentrasi hormon testosteron dalam darah,

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sebaliknya konsumsi energi atau lemak menyebabkan meningkatnya konsentrasi hormon testosteron dalam darah.

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Kesimpulan
Pemberian tepung ikan dalam konsentrat menghasilkan pertambahan bobot badan dan efisiensi pakan lebih tinggi, profil darah lebih baik, penampilan tingkah laku seksual dan produksi semen lebih baik dibanding pemberian bungkil kedele plus urea dalam konsentrat. Pemberian tepung ikan lokal dalam konsentrat menghasilkan pertambahan bobot badan dan konversi pakan lebih baik, tetapi menghasilkan penampilan tingkah laku seksual dan produksi semen yang tidak berbeda nyata dengan pemberian tepung ikan impor dalam konsentrat.

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Formatted: Font: 10 pt, Font color: Auto, Indonesian, Character scale: 98%, Expanded by 0,3 pt Formatted Formatted ... [87] ... [88]

Deleted: Anonymous, 2002. Peruvian fishmeal : your ingredient to profitability in the livestock industry. Aquaculture & Fisheries Expo Lima, Peru. September 2002 Anonymous, 1999. Dietary manipulation to increase fertility. Description of Corresponding Document : WO9966877. www.esp@cenet description view.htm. Anonymous. 2001a. Fish meal, Menhaden. Ingredients 101.com. P.O. Box 420, Grafton, WI 53024. Anonymous. 2001b. Testosterone, Catalog no. 2095. Diagnostic Automation, Inc. 23961 Craftsman Rd., Suite E/F. Calabasas, C.A. 91302. Aryawansa, S. 2000. The evaluation of functional properties of fish meal. Final Project 2000. The United Nations University.Fisheries Training Programme. Reykjavik. Iceland. Blauwiekel, R., S. Xu, and J. H. Harrison. 1992. The use of cereal grains and by-product feeds to meet the amino acid requirements of dairy cattle. In: Proc. 27th Pacific Northwest Animal Nutrition Conference. Spokane, WA. pp 225236. ... [89] Formatted Formatted Formatted ... [90] ... [91] ... [92]

Page 3: [1] Deleted

5/29/2007 5:10:00 PM

. Setiap kali pengambilan sampel darah dilakukan


Page 3: [2] Deleted x 5/29/2007 5:11:00 PM

dalam selang waktu satu jam (


Page 3: [3] Formatted x 5/29/2007 5:07:00 PM

Font: 11 pt, Italic, Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 3: [4] Deleted x 5/29/2007 5:12:00 PM

Sampel darah diambil dari vena jugularis sebanyak 3 ml per ekor dengan menggunakan spuit 5 ml yang dilengkapi jarum nomor 21.
Page 3: [5] Formatted x 5/29/2007 5:07:00 PM

Font: 11 pt, Italic, Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 3: [6] Formatted x 5/29/2007 5:07:00 PM

Font: 11 pt, Italic, Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [7] Deleted x 5/29/2007 5:17:00 PM

(n = 280) *
Page 5: [7] Deleted x 5/29/2007 5:17:00 PM

(n = 280)
Page 5: [7] Deleted x 5/29/2007 5:17:00 PM

(n = 280)
Page 5: [8] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [9] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [10] Formatted x 5/30/2007 9:48:00 AM

Indent: Left: 0 cm, Right: 0 cm


Page 5: [11] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [11] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [11] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [11] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Not Superscript/ Subscript, Character scale: 98%, Expanded by 0,3 pt


Page 5: [11] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [11] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [11] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [11] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [11] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt

Page 5: [11] Formatted

5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [11] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [12] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [13] Formatted x 5/30/2007 9:48:00 AM

Indent: Left: 0 cm
Page 5: [14] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [14] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [14] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [14] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [14] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [14] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [14] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [15] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [16] Formatted x 5/30/2007 9:48:00 AM

Indent: Left: 0 cm
Page 5: [17] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [17] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [17] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [17] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [17] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [17] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [17] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [17] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt

Page 5: [18] Formatted

5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [19] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [19] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [19] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [19] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [19] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [19] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [19] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [19] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [20] Formatted x 5/30/2007 9:48:00 AM

Indent: Left: 0 cm
Page 5: [21] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [22] Formatted x 5/30/2007 9:48:00 AM

Indent: Left: 0 cm
Page 5: [23] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [23] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [23] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [23] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [23] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [23] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [23] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [24] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [25] Formatted x 5/30/2007 9:48:00 AM

Indent: Left: 0 cm

Page 5: [26] Formatted

5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [26] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [26] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [26] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [26] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [26] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [26] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [26] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [27] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [28] Formatted x 5/30/2007 9:48:00 AM

Indent: Left: 0 cm, Right: 0 cm


Page 5: [29] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [29] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [29] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [29] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [29] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [29] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [29] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [29] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [30] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [30] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [30] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt

Page 5: [30] Formatted

5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [30] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [30] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [31] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [32] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [32] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [32] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [32] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [32] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [32] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [32] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [33] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [33] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [33] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [33] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [33] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [33] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [33] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [33] Formatted x 5/30/2007 9:42:00 AM

Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [34] Formatted x 5/29/2007 5:25:00 PM

Font: 10 pt, Italic, Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 5: [35] Formatted x 5/30/2007 9:52:00 AM

Indent: Left: 0 cm

Page 5: [36] Formatted

5/30/2007 9:52:00 AM

Indent: Left: 0 cm
Page 5: [37] Formatted x 5/30/2007 9:52:00 AM

Indent: Left: 0 cm
Page 5: [38] Formatted x 5/30/2007 9:52:00 AM

Indent: Left: 0 cm
Page 5: [39] Formatted x 5/30/2007 9:52:00 AM

Indent: Left: 0 cm
Page 5: [40] Formatted x 5/30/2007 9:52:00 AM

Indent: Left: 0 cm
Page 5: [41] Formatted x 5/30/2007 9:52:00 AM

Indent: Left: 0 cm
Page 5: [42] Formatted x 5/30/2007 9:52:00 AM

Indent: Left: 0 cm
Page 5: [43] Formatted x 5/30/2007 9:52:00 AM

Indent: Left: 0 cm
Page 5: [44] Deleted x x x 5/29/2007 3:49:00 PM 5/29/2007 3:50:00 PM 5/29/2007 3:56:00 PM

yang
Page 5: [44] Deleted

yang
Page 6: [45] Deleted

sperma
Page 6: [45] Deleted x 5/29/2007 3:56:00 PM

sperma
Page 6: [45] Deleted x 5/29/2007 5:45:00 PM

bisa
Page 6: [45] Deleted x 5/29/2007 5:44:00 PM

sel telur
Page 6: [45] Deleted x 5/29/2007 3:56:00 PM

sperma
Page 6: [45] Deleted x 5/29/2007 3:56:00 PM

sperma
Page 6: [45] Deleted x 5/29/2007 5:43:00 PM

hasil pengamatan
Page 6: [45] Deleted x 5/29/2007 5:46:00 PM

pada
Page 6: [45] Deleted x 5/29/2007 3:56:00 PM

sperma
Page 6: [46] Deleted x 5/29/2007 5:42:00 PM

Page 6: [46] Deleted

5/29/2007 5:36:00 PM

tinggi
Page 6: [47] Deleted x 5/29/2007 5:46:00 PM

l
Page 6: [47] Deleted x 5/29/2007 3:56:00 PM

sperma
Page 6: [47] Deleted x 5/29/2007 5:47:00 PM

,
Page 6: [48] Deleted x 5/29/2007 5:47:00 PM

l
Page 6: [48] Deleted x 5/29/2007 3:56:00 PM

sperma
Page 6: [48] Deleted x 5/29/2007 5:48:00 PM

dibanding
Page 6: [48] Deleted x 5/29/2007 5:48:00 PM

kemudian
Page 6: [48] Deleted x 5/29/2007 5:49:00 PM

Page 6: [48] Deleted

5/29/2007 5:52:00 PM

Page 6: [49] Deleted

5/29/2007 5:53:00 PM

,
Page 6: [49] Deleted x 5/29/2007 5:53:00 PM

dan motilitas sperma minimal pada semen setelah dibekukan dan dicairkan adalah 30 %.
Page 6: [49] Deleted x 5/29/2007 3:56:00 PM

sperma
Page 6: [49] Deleted x 5/29/2007 3:56:00 PM

sperma
Page 6: [49] Deleted x 5/29/2007 3:56:00 PM

sperma
Page 6: [49] Deleted x 5/29/2007 3:56:00 PM

sperma
Page 6: [49] Deleted x 5/29/2007 3:56:00 PM

sperma
Page 6: [49] Deleted x 5/29/2007 5:54:00 PM

pada
Page 6: [49] Deleted x 5/29/2007 5:54:00 PM

sperma per ml semen


Page 6: [49] Deleted x 5/29/2007 5:55:00 PM

sperma per ml semen


Page 6: [50] Deleted x 5/29/2007 3:56:00 PM

sperma
Page 6: [50] Deleted x 5/29/2007 5:41:00 PM

/
Page 6: [51] Deleted x 5/29/2007 3:56:00 PM

sperma
Page 6: [51] Deleted x 5/29/2007 3:56:00 PM

sperma
Page 6: [51] Deleted x 5/29/2007 3:56:00 PM

sperma
Page 6: [52] Deleted x 5/29/2007 3:56:00 PM

sperma
Page 6: [52] Deleted x 5/29/2007 3:56:00 PM

sperma
Page 6: [52] Deleted x 5/29/2007 5:45:00 PM

pada
Page 9: [53] Formatted x 5/30/2007 11:28:00 AM

Font: 10 pt, Font color: Auto, Indonesian, Character scale: 98%, Expanded by 0,3 pt
Page 9: [53] Formatted x 5/30/2007 11:28:00 AM

Font: 10 pt, Font color: Auto, Indonesian, Character scale: 98%, Expanded by 0,3 pt
Page 9: [53] Formatted x 5/30/2007 11:28:00 AM

Indonesian, Character scale: 98%


Page 9: [54] Formatted x 5/30/2007 11:28:00 AM

Font: 10 pt, Font color: Auto, Indonesian, Character scale: 98%, Expanded by 0,3 pt
Page 9: [54] Formatted x 5/30/2007 11:28:00 AM

Font: 10 pt, Font color: Auto, Indonesian, Character scale: 98%, Expanded by 0,3 pt
Page 9: [54] Formatted x 5/30/2007 11:28:00 AM

Font: 10 pt, Font color: Auto, Indonesian, Character scale: 98%, Expanded by 0,3 pt
Page 9: [54] Formatted x 5/30/2007 11:28:00 AM

Font: 10 pt, Font color: Auto, Indonesian, Character scale: 98%, Expanded by 0,3 pt
Page 9: [54] Formatted x 5/30/2007 11:28:00 AM

Font: 10 pt, Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 9: [55] Deleted x 5/29/2007 6:11:00 PM

p
Page 9: [55] Deleted x 5/29/2007 3:56:00 PM

sperma
Page 9: [55] Deleted x 5/29/2007 3:56:00 PM

sperma
Page 9: [56] Formatted x 5/30/2007 11:28:00 AM

Font: 10 pt, Font color: Auto, Indonesian, Character scale: 98%, Expanded by 0,3 pt
Page 9: [56] Formatted x 5/30/2007 11:28:00 AM

Font: 10 pt, Font color: Auto, Indonesian, Character scale: 98%, Expanded by 0,3 pt
Page 9: [56] Formatted x 5/30/2007 11:28:00 AM

Font: 10 pt, Font color: Auto, Indonesian, Character scale: 98%, Expanded by 0,3 pt
Page 9: [57] Formatted x 5/30/2007 11:28:00 AM

Font: 10 pt, Font color: Auto, Indonesian, Character scale: 98%, Expanded by 0,3 pt
Page 9: [57] Formatted x 5/30/2007 11:28:00 AM

Font: 10 pt, Font color: Auto, Indonesian, Character scale: 98%, Expanded by 0,3 pt
Page 9: [57] Formatted x 5/30/2007 11:28:00 AM

Font: 10 pt, Font color: Auto, Indonesian, Character scale: 98%, Expanded by 0,3 pt
Page 9: [58] Formatted x 5/30/2007 11:28:00 AM

Font: 10 pt, Font color: Auto, Indonesian, Character scale: 98%, Expanded by 0,3 pt
Page 9: [58] Formatted x 5/30/2007 11:28:00 AM

Font: 10 pt, Font color: Auto, Indonesian, Character scale: 98%, Expanded by 0,3 pt
Page 9: [58] Formatted x 5/30/2007 11:28:00 AM

Font: 10 pt, Font color: Auto, Indonesian, Character scale: 98%, Expanded by 0,3 pt
Page 9: [58] Formatted x 5/30/2007 11:28:00 AM

Font: 10 pt, Font color: Auto, Indonesian, Character scale: 98%, Expanded by 0,3 pt
Page 9: [58] Formatted x 5/30/2007 11:28:00 AM

Font: 10 pt, Indonesian, Character scale: 98%, Expanded by 0,3 pt


Page 9: [59] Formatted x 5/30/2007 11:28:00 AM

Font: 10 pt, Font color: Auto, Indonesian, Character scale: 98%, Expanded by 0,3 pt
Page 9: [59] Formatted x 5/30/2007 11:28:00 AM

Font: 10 pt, Font color: Auto, Indonesian, Character scale: 98%, Expanded by 0,3 pt
Page 9: [59] Formatted x 5/30/2007 11:28:00 AM

Font: 10 pt, Font color: Auto, Indonesian, Character scale: 98%, Expanded by 0,3 pt
Page 9: [59] Formatted x 5/30/2007 11:28:00 AM

Font: 10 pt, Font color: Auto, Indonesian, Character scale: 98%, Expanded by 0,3 pt
Page 9: [60] Formatted x 5/30/2007 11:28:00 AM

Font: 10 pt, Font color: Auto, Indonesian, Character scale: 98%, Expanded by 0,3 pt
Page 9: [60] Formatted x 5/30/2007 11:28:00 AM

Font: 10 pt, Font color: Auto, Indonesian, Character scale: 98%, Expanded by 0,3 pt
Page 9: [60] Formatted x 5/30/2007 11:28:00 AM

Font: 10 pt, Font color: Auto, Indonesian, Character scale: 98%, Expanded by 0,3 pt
Page 9: [60] Formatted x 5/30/2007 11:28:00 AM

Font: 10 pt, Not Bold, Font color: Auto, Indonesian, Character scale: 98%, Expanded by 0,3 pt
Page 9: [61] Formatted x 5/30/2007 11:28:00 AM

Font: 10 pt, Font color: Auto, Indonesian, Character scale: 98%, Expanded by 0,3 pt
Page 9: [61] Formatted x 5/30/2007 11:28:00 AM

Font: 10 pt, Font color: Auto, Indonesian, Character scale: 98%, Expanded by 0,3 pt
Page 9: [61] Formatted x 5/30/2007 11:28:00 AM

Font: 10 pt, Font color: Auto, Indonesian, Character scale: 98%, Expanded by 0,3 pt
Page 9: [62] Formatted x 5/30/2007 11:28:00 AM

Font: 10 pt, Character scale: 98%, Expanded by 0,3 pt


Page 9: [62] Formatted x 5/30/2007 11:28:00 AM

Font: 10 pt, Character scale: 98%, Expanded by 0,3 pt


Page 9: [62] Formatted x 5/30/2007 11:28:00 AM

Font: 10 pt, Italic, Character scale: 98%, Expanded by 0,3 pt

Page 9: [62] Formatted

5/30/2007 11:28:00 AM

Font: 10 pt, Character scale: 98%, Expanded by 0,3 pt


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Font: 10 pt, Character scale: 98%, Expanded by 0,3 pt


Page 9: [63] Formatted x 5/30/2007 11:28:00 AM

Font: 10 pt, Font color: Auto, Indonesian, Character scale: 98%, Expanded by 0,3 pt
Page 9: [63] Formatted x 5/30/2007 11:28:00 AM

Font: 10 pt, Font color: Auto, Indonesian, Character scale: 98%, Expanded by 0,3 pt
Page 9: [63] Formatted x 5/30/2007 11:28:00 AM

Font: 10 pt, Font color: Auto, Indonesian, Character scale: 98%, Expanded by 0,3 pt
Page 9: [63] Formatted x 5/30/2007 11:28:00 AM

Font: 10 pt, Font color: Auto, Indonesian, Character scale: 98%, Expanded by 0,3 pt
Page 9: [63] Formatted x 5/30/2007 11:28:00 AM

Font: 10 pt, Font color: Auto, Indonesian, Character scale: 98%, Expanded by 0,3 pt
Page 9: [64] Formatted x 5/30/2007 11:28:00 AM

Font: 10 pt, Font color: Auto, Character scale: 98%, Expanded by 0,3 pt
Page 9: [64] Formatted x 5/30/2007 11:28:00 AM

Font: 10 pt, Font color: Auto, Character scale: 98%, Expanded by 0,3 pt
Page 9: [64] Formatted x 5/30/2007 11:28:00 AM

Font: 10 pt, Not Italic, Font color: Auto, Character scale: 98%, Expanded by 0,3 pt
Page 9: [64] Formatted x 5/30/2007 11:28:00 AM

Font: 10 pt, Font color: Auto, Character scale: 98%, Expanded by 0,3 pt
Page 9: [64] Formatted x 5/30/2007 11:28:00 AM

Font: 10 pt, Font color: Auto, Character scale: 98%, Expanded by 0,3 pt
Page 9: [65] Formatted x 5/30/2007 11:28:00 AM

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Anonymous, 2002. Peruvian fishmeal : your ingredient to profitability in the livestock industry. Aquaculture & Fisheries Expo Lima, Peru. September 2002 Anonymous, 1999. Dietary manipulation to increase fertility. Description of Corresponding Document : WO9966877. www.esp@cenet description view.htm. Anonymous. 2001a. Fish meal, Menhaden. Ingredients 101.com. P.O. Box 420, Grafton, WI 53024. Anonymous. 2001b. Testosterone, Catalog no. 2095. Diagnostic Automation, Inc. 23961 Craftsman Rd., Suite E/F. Calabasas, C.A. 91302. Aryawansa, S. 2000. The evaluation of functional properties of fish meal. Final Project 2000. The United Nations University.Fisheries Training Programme. Reykjavik. Iceland.

Blauwiekel, R., S. Xu, and J. H. Harrison. 1992. The use of cereal grains and by-product feeds to meet the amino acid requirements of dairy cattle. In: Proc. 27th Pacific Northwest Animal Nutrition Conference. Spokane, WA. pp 225-236. Burke, J.M., C.R. Staples, C.A. Risco, R.L. De La Sota, and W.W. Thatcher. 1997. Effect of feeding a ruminant grade Manhaden fish meal on reproductive and productive performance of lactating dairy cows. J. Dairy Sci. 80 : 3386. Delgadillo J.A., M. E. Cortez, G. Duarte, P. Chemineau, and B. Malpaux. 2004. Evidence that the photoperiod controls the annual changes in testosterone secretion, testicular and body weight in subtropical male goats. Reprod. Nutr. Dev. 44 (2004) 183193. INRA, EDP Sciences, 2004. Delgadillo, J. A., J. A. Flores, F. G. Vliz, H. F. Hernndez, G. Duarte, J. Vielma, P. Poindron, P. Chemineau and B. Malpaux. 2002. Induction of sexual activity in lactating anovulatory female goats using male goats treated only with artificially long days. J. Anim. Sci. 80 : 2780-2786. Estienne, J.M. and A.F. Harper, 2004. Boar feeding and nutrition. Livestock Update, October 2004. Virginia Cooperative Extension. Virginia Polytechnic Institute and State University. FIN, Fishmeal Information Network. 1999. Feeding fish meal improves cow fertility. FIN Nutritional Paper 4. 4 The Forum, Minerva Business Park, Peterborough, Cambridgeshire. PE2 6FT, UK. FIN, Fishmeal Information Network. 2000b. Feed fishmeal to growing beef cattle for cost-effective extra weight gain and leaner carcasses. FIN Press Release-12, 26 June 2000. 4 The Forum, Minerva Business Park, Peterborough, Cambridgeshire. PE2 6FT, UK. Grigsby, K. N., F. M. Rouquette, Jr., W. C. Ellis, and D. P. Hutche-son. 1999. Use of self-limiting fish meal and corn supplements for calves grazing rye-ryegrass pastures. Journal of Production Agriculture, 4 : 476. Grummer, R.R. and D.J. Carroll. 1991. Effects of dietary fat on metabolic disorders and reproductive performance of dairy cattle. J. Anim. Sci. 69 : 3838. Haenlein, G.F.W., R. Caccese, and M.C. Smith. 2006. Artificial insemination. Artificial Insemination (AI) - Goats & Health - goatworld_co.htm. Herwijanti, E. 2004. Pengaruh tingkah laku seksual terhadap kualitas semen pada berbagai bangsa sapi potong. Tesis. Program Pascasarjana. Universitas Brawijaya Malang. 2004. Hullet, C.V. and M. Shelton. 1987. Reproductive cycle of sheep and goat. Edited by E.S.E. Hafez. Reproduction in Farm Animal. 5 th edition. Lea and Febiger. Philadelphia. Hussein, H.S., and R.M. Jordan. 1991. Fish meal as a protein supplement in ruminant diets. A Review. J. Anim. Sci. 62 : 2147. Kheradmand, A., H. Babaei, and R.A. Batavani. 2006. Effect of improved diet on semen quality and scrotal circumference in the ram. Vet. Archive 76, 333-341, 2006. Kohn R. A., M. M. Dinneen and E. Russek-Cohen. 2005. Using blood urea nitrogen to predict nitrogen excretion and efficiency of nitrogen utilization in cattle, sheep, goats, horses, pigs, and rats. J. Anim. Sci. 83 : 879-889. Kridli, R.T., A.Y. Abdullah and M. Momani Shaker. 2006. Sexual performance and reproductive characteristics of young adult awassi, charollais-awassi and romanov-awassi rams. Sheep & Goat Research Journal, Volume 21, 2006:12-16 Lotthammer, K.H. 1991. Influence of nutrition on reproductive performance of the milking/gestating cow in the tropics. In Feeding dairy cows in the tropics. Proceedings of the FAO Expert Consultation held in Bangkok, Thailand, 711 July 1989. Edited by Andrew Speedy and Ren

Sansoucy. Food and Agriculture Organization of The United Nations. FAO Animal Production and Health Paper 86, Rome, 1991. Louis, G.F., A.J. Lewis, W.C. Weldon, P.M. Ermer, P.S. Miller, R.J. Kittok, and W.W. Stroup, 1994b. The effect of energy and protein intakes on boar libido, semen characteristics, and plasma hormone concentration. J. Anim. Sci. 72 : 2051-2060. Louis, G.F., A.J. Lewis, W.C. Weldon, P.S. Miller, R.J. Kittok, and W.W. Stroup, 1994a. The effect of energy and protein intakes on boar libido, semen characteristics, and plasma hormone concentration. J. Anim. Sci. 72 : 2038-2050. Marjuki. 2007. Evaluasi kualitas tepung ikan produksi dalam negeri dibanding tepung ikan impor. Jurnal Agritek Volume 15 No.1. Oglle, T.F. and A. Costoff. 2001. Endocrinology, male reproductive physiology. In essentials of human physiology. A Breakthough in Human Physiology Education. Edited by Tomas M. Nosek. Oyeyemi, M. O., M. O. Akusu, and O. E. Ola-Davies. 2000. Effect of successive ejaculations on the spermiogram of west african dwarf goats (capra hircus l.). Veterinary Archive 70 : 215-221. Pike, I.H., E.L. Miller, and K. Short. 1994. The role of fishmeal in dairy cow nutrition. IFOMA Technical Bulletin 27 (August 1994). IFOMA, St Albans, Hertfordshire, UK. Rivas, A.L. and J.S. Serrato-Corona. 2005. Effects of undegradable intake protein on milk yield, dry matter digestibility and blood urea nitrogen in lactating goats fed ammoniated corn stover. Proceedings, Western Section, American Society of Animal Science Vol. 56, 2005. Robinson, J.J., C.J. Ashworth, J.A. Rooke, L.M. Mitchell, and T.G. McEvoy. 2005. Nutrition and fertility in ruminant livestock. Elsevier B.V. All rights reserved. Rocha, A., M. Carpena, B. Triplett, D. W. Forrest, and R. D. Randel. 1995. Effect of ruminally undegradable protein from fish meal on growth and reproduction of peripuberal brahman bulls. Journal of Animal Science, 73 : 947-953. Salisbury, G.W., van Demark, N.L. & Codge, J.B. 1978. Physiology of reproduction and artificial insemination of cattle. 2nd ed. San Francisco, W.H. Freeman & Co. Schroeder, J.W. 1999. By-products and regionally available alternative feedstuffs for dairy cattle. North Dakota State University (NDSU) Extension Service. Bulletin AS-1180. Veira, D. M,, J. G. Proulx, G. Buttler, and A. Fortin. 1988. Utilization of grass silage by cattle: Further observations on the effect of fish meal. Canadian J. Anim. Sci. 68 : 1225. Volek, J.S., W.J. Kraemer, J.A. Bush, T. Incledon, and M. Boetes. 1997. Testosterone and cortisol in relationship to dietary nutrients and resistance exercise. J. Appl. Phys. 82 (1) : 49-54. 1997. Walkden-Brown, S.W., B.J. Restall and W.A., Taylor. 2006. Testicular and epididymal sperm content in grazing Cashmere bucks: seasonal variation and prediction from measurements in vivo. Reproduction, Fertility and Development 6(6) 727 736. Wildeus, S. 2000. Reproduction and breeding. Goat Research. Agricultural Research and Extension Programs. Langston University. Langston, UK. Zinn, R.A. and F.N. Owens. 1993. Ruminal escape protein for lightweight feedlot calves. J. Anim. Sci. 71 : 1677.

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ANIMAL PRODUCTION, September 2007, hlm. 145 152 ISSN 1411 2027 Terakreditasi No.56/DIKTI/Kep/2005

Vol. 9 No.3

Penentuan Waktu Ekuilibrasi pada Pembekuan Semen Kuda Menggunakan Bahan Pengencer Susu Skim
(Determination of Equilibration Time of Stallion Semen Freezing with Extender Skim Milk)
Raden Iis Arifiantini*, Iman Supriatna dan Samsurizal
Fakultas Kedokteran Hewan, Institut Pertanian Bogor Jl. Agatis Kampus IPB Darmaga, Bogor 16680

ABSTRACT: The objectives of this experiment were to determine the best equilibration time with extender skim milk in order to maintain the quality of frozen stallion semen. The semen was collected from three 5-8 years old stallions using artificial vagina. The semen characteristics and quality were evaluated macro- and microscopically and extended with skim milk extender (1:1). The semen was centrifuged at 3000 rpm for 15 minutes. Supernatant was removed and the pellet was re-extended with skim milk extender with 5% glycerol. The extended semen then packed in mini straw (0.5 mL), equilibrated at 5oC for 1 hr (E1), 2 hrs (E2 ) and 3 hrs (E3), frozen in the liquid nitrogen (N2) vapor for 15 minutes and then stored in liquid N2 container for 24 hrs. The straw than thawed at 37oC for 30 seconds. The percentages of sperm motility and viability were observed. The result of this research showed that there were no significant differences between equilibration at 1, 2 and 3 hrs on the percentages of sperm motility with the average of post thawed motility was 23.60 7.10%. The equilibration time affected the sperm viability (P<0.05) in all stallions, with the percentages of viable sperm between 38.10 % and 46.50%. Key Words: Equilibration, frozen semen, stallion

Pendahuluan
Waktu ekuilibrasi adalah periode yang diperlukan spermatozoa sebelum pembekuan untuk menyesuaikan diri dengan pengencer supaya pada saat pembekuan kematian spermatozoa yang berlebihan dapat dicegah. Waktu ekuilibrasi didefinisikan sebagai waktu yang dibutuhkan oleh krioprotektan untuk mencapai keseimbangan pada kedua sisi membran plasma (Bearden et al., 2004). Pada saat ekuilibrasi spermatozoa akan beradaptasi dengan pengencernya, sehingga dapat menurunkan persentase mortalitas (kematian) spermatozoa pada saat pembekuan. Menurut Kacker dan Panwar (1996), ekuilibrasi bertujuan melindungi spermatozoa dari kematian yang disebabkan oleh penurunan tekanan osmotik yang mengganggu keutuhan membran spermatozoa akibat pembekuan. Waktu ekuilibrasi pada semen berbagai ternak berbeda-beda. Menurut Bearden et al. (2004) pada pembekuan semen domba, waktu ekuilibrasi pada

suhu 5 o C terbaik adalah 2 jam dan 4 jam pada rusa. Pada semen sapi Friesian Holstein (FH) (Arifiantini et al., 2005) juga melakukan ekuilibrasi selama 4 jam dengan suhu 5 o C. Han et al. (2005) membandingkan waktu ekuilibrasi pada pembekuan semen itik dengan waktu 10 dan 60 menit, ternyata ekuilibrasi selama 10 menit menunjukkan kualitas semen setelah thawing lebih baik dibandingkan 60 menit. Pada pembekuan semen kambing Peranakan Etawah (PE) ekuilibrasi selama 4 jam pada suhu 35 o C lebih baik dibandingkan dengan 1, 2 ataupun 3 jam (Raya, 2000). Untuk semen kuda, Arruda et al. (2002) melakukan ekuilibrasi pada suhu 5 oC selama 2 jam dengan krioprotektan gliserol dan etilen glikol 5%, sedangkan Medeiros et al. (2002) dengan waktu ekuilibrasi selama 1 jam pada suhu yang sama. Oleh karena itu, penelitian ini bertujuan untuk menentukan waktu ekuilibrasi terbaik (1, 2 dan 3 jam) pada pembekuan semen kuda menggunakan pengencer susu skim.

Metode Penelitian
Semen yang digunakan dalam penelitian ini berasal dari tiga ekor kuda jantan yang sudah mencapai sexual maturity (antara 5 dan 8 tahun), terdiri atas: satu ekor generasi empat (G4)

Penulis Korespondensi e-mail: Iis_arifiantini@telkom.net

145

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Thoroughbred (Spirit of Jatim; SJ), satu ekor American Pinto (Sony Boy; SB) dan satu ekor Warmblood (Prince Couperos; PC) dalam kondisi sehat dan terseleksi melalui pengujian daily sperm output (produksi spermatozoa harian) dan satu ekor kuda betina sebagai pemancing, seluruhnya milik Athena Stable Depok. Bahan pengencer dasar yang digunakan untuk sentrifugasi adalah susu skim glukosa, dengan komposisi sebagai berikut: susu skim 2,4 g dan glukosa 4,0 g (Kenney et al., 1975) dalam akuades ad 100 ml, dipanaskan pada suhu 92-95 oC selama 10 menit, didinginkan kemudian ditambah antibiotika streptomisin 100 mg dan penisilin 100 000 IU. Pengencer untuk semen beku adalah bahan pengencer dasar ditambah gliserol 5 %. Penampungan semen kuda dilakukan dua kali dalam satu minggu menggunakan vagina buatan tipe Nishikawa dengan tabung penampung modifikasi tipe Missouri (Arifiantini et al., 2006). Semen segar yang diperoleh dievaluasi terhadap volume, warna, konsistensi dan derajat keasaman (pH). Secara mikroskopis dievaluasi persentase spermatozoa motil, viabilitas, konsentrasi dan morfologi spermatozoa. Motilitas spermatozoa dinilai dengan mikroskop cahaya listrik (Olympus CH20) dengan pembesaran 400 x secara subjektif kuantitatif (Mickelsen et al., 1993). Viabilitas spermatozoa dilakukan dengan pewarnaan eosin nigrosin (MacPherson, 2001). Konsentrasi spermatozoa (x10 6 mL-1 ) menggunakan Neubauer chamber dengan pengenceran 100 kali dalam NaCl 3 %, dan morfologi spermatozoa (normalitas) dilakukan dengan pewarnaan Williams. Semen yang mempunyai persentase motilitas >65% dengan konsentrasi >200 x10 6 mL-1 dengan morfologi spermatozoa normal >70%, diencerkan dengan susu skim (1:1). Disentrifugasi (Hettrich, EBA 3S Tuttlingen) dengan kecepatan 3000 rpm selama 15 menit. Supernatan dibuang dan pellet (spermatozoa) diencerkan kembali dengan pengencer susu skim yang mengandung gliserol 5 % dengan konsentrasi spermatozoa 200 X 10 6 mL -1 . Semen tersebut kemudian dikemas ke dalam straw 0.5 mL. Diekuilibrasi pada suhu 4-5 o C masingmasing selama 1 jam (E1 ), 2 jam (E2 ) dan 3 jam (E3 ). Kemudian dibekukan pada uap N 2 cair (suhu -150 o C) selama 10 menit, disimpan dalam kontainer N 2 cair (suhu -196 o C). Keberhasilan pembekuan dievaluasi 24 jam kemudian. Semen beku di-thawing pada suhu 37 oC selama 30 detik dengan menggunakan water bath (Chenier et al.,

1998). Parameter yang diukur dalam penelitian ini adalah persentase motilitas dan viabilitas spermatozoa. Persentase spermatozoa yang berhasil pulih dari proses pembekuan yang disebut recovery rate (RR) dihitung dengan cara membandingkan motilitas spermatozoa setelah pembekuan dengan motilitas semen segar dikalikan 100% (Han et al., 2005).
Persentase spermatozoa motil setelah thawing
RR= Persentase spermatozoa motil semen segar x 100 %

Rancangan percobaan yang digunakan adalah rancangan acak kelompok (RAK) dan untuk masingmasing pejantan diulang sebanyak empat kali. Data yang diperoleh dianalisis dengan analisis variansi dan dilanjutkan dengan uji jarak berganda Duncan (Steel dan Torrie, 1995).

Hasil dan Pembahasan


Kualitas Semen Segar
Hasil dari evaluasi semen yang telah dilakukan menunjukkan kualitas yang cukup baik. Volume semen segar sebesar 27,709,80 mL, berwarna putih keruh dengan konsistensi encer dan pH 7,100,10. Secara mikroskopis persentase spermatozoa motil progresif adalah 67,17,20%, viabilitas 78,803,30%, spermatozoa normal 73,604,90% dengan konsentrasi 222,9017,30x10 6 mL-1 . Secara individu dari ketiga kuda jantan yang digunakan SJ menunjukkan kulitas semen cair yang paling rendah dibandingkan dengan PC dan SB (Tabel 1). Karakteristik semen segar tersebut secara keseluruhan masih dalam kisaran normal untuk kuda (Brinsko et al., 2000; Quintero-Moreno et al., 2003). Tingginya standar deviasi menunjukkan besarnya variasi volume semen dari ketiga pejantan yang berbeda bangsa. Menurut Dowsett dan Knott (1996) bangsa dan umur kuda mempengaruhi karakteristik semen yang dihasilkan.

Pengaruh Lama Ekuilibrasi Persentase Motilitas dan Spermatozoa setelah Thawing

terhadap Viabilitas

Waktu ekuilibrasi yang optimum dalam pembekuan semen pada berbagai ternak bervariasi. Pada pembekuan semen kuda hasil penelitian ini, lama ekuilibrasi berbeda pengaruhnya untuk tiap individu. Pada jantan SJ, perlakuan E1 , E2 dan E3 tidak menunjukkan perbedaan motilitas setelah

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Tabel 1. Karakteristik semen cair kuda Evaluasi Spirit of Jatim Makroskopis Volume (mL) 25,0 6,10 Warna putih keruh pH 7,10 0,10 Konsistensi encer Mikroskopis Motilitas spermatozoa (%) 57,50 2,50 Viabilitas spermatozoa (%) 78,90 2,00 Normalitas spermatozoa (%) 71,80 3,50 Konsentrasi (x10 6 mL -1 ) 207,00 12,80
80.0 70.0 60.0

Nama kuda Sony Boy 22,50 2,50 putih keruh 7,10 0,10 encer 71,30 2,20 77,70 3,30 75,30 2,40 227,30 15,90

Prince Couperous 36,25 11,40 putih keruh 6,95 0,20 encer 72,50 2,50 79,80 4,00 76,10 1,40 234,50 8,60

Motilitas (%)

50.0 40.0 30.0 20.0 10.0 0.0 SS SE ST

Viabilitas (%)

1 Jam 2 Jam 3 Jam

SS

SE

ST

Nama Kuda

Nama Kuda

Gambar 1. Persentase motilitas dan viabilitas spermatozoa pada berbagai tahapan pengolahan semen untuk Spirit of Jatim (SS = semen segar; SE = setelah ekuilibrasi; ST = setelah thawing)

thawing dengan motilitas rata-rata 17,30% (P>0,05), sedangkan viabilitas spermatozoa setelah thawing pada jantan SJ, ternyata E3 (47,80%) menunjukkan persentase yang lebih tinggi dibanding E2 (45,30%), dan E1 (38,40%) (Gambar 1). Pada kuda SB dan PC, persentase spermatozoa motil setelah thawing masing-masing (30,00%; 28,90%) dan 3 jam (28,30% ; 30,60%) lebih tinggi dibandingkan E1 (20,60%; 23,30%) (P<0,05). Persentase viabilitas spermatozoa setelah thawing pada PC tertinggi pada E3 (50,00%) diikuti oleh E2 (44,60%), sedangkan E1 menunjukkan nilai yang lebih rendah yaitu 37,30%. Untuk kuda SB, E2 menunjukkan persentase viabilitas spermatozoa 49,70%, lebih tinggi dibandingkan E1 (40,10%) dan E3 (38,40%) (Gambar 2 dan 3). Secara keseluruhan lama ekuilibrasi ini tidak mempengaruhi persentase motilitas setelah thawing dari ketiga kuda yang digunakan (P>0,05) dengan persentase motilitas setelah thawing 23,60 7,10% (Gambar 4). Lama ekuilibrasi mempengaruhi persentase viabilitas spermatozoa baik secara individu atau secara keseluruhan. Waktu ekuilibrasi selama 2 dan 3 jam menunjukkan persentase viabilitas spermatozoa lebih baik dibandingkan dengan 1 jam.

Penurunan kualitas dari semen segar ke setelah ekuilibrasi masih rendah, untuk motilitas spermatozoa turun antara 4,40% dan 7,50%, sedangkan untuk viabilitas spermatozoa turun antara 1,80% - 11,10%. Dari setelah ekuilibrasi ke setelah thawing penurunan kualitas terjadi cukup tinggi yaitu berkisar antara 30,70% dan 45,60% untuk motilitas spermatozoa dan 18,80% - 38,20% untuk viabilitas spermatozoa. Sehingga total penurunan kualitas yang terjadi mulai semen segar ke setelah thawing antara 37,50% dan 50,60% untuk motilitas spermatozoa dan 28,60% - 42,50% untuk viabilitas spermatozoa (Tabel 2). Keberhasilan pembekuan semen tidak hanya dinilai dari persentase motilitas setelah thawing, hal ini disebabkan oleh motilitas semen segar yang berbeda pada ketiga kuda yang digunakan. Persentase spermatozoa yang dapat pulih kembali setelah pembekuan disebut dengan recovery rate (RR) yang memberikan gambaran keberhasilan dari proses pembekuan itu sendiri. Tanpa melihat lama ekuilibrasi yang dilakukan, secara keseluruhan ternyata PC mempunyai nilai RR tertinggi yaitu 38,10% diikuti oleh SB 36,90% dan SJ sebesar 29,60% (Gambar 5).

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80.0 60.0
Viabilitas (%)

1 Jam 2 Jam 3 Jam

40.0 20.0 0.0 SS SE Nama Kuda ST

SS

SE Nama Kuda

ST

Gambar 2. Persentase motilitas dan viabilitas spermatozoa pada berbagai tahapan pengolahan semen untuk Sony boy (SS=semen segar; SE= Setelah ekuilibrasi; ST = Setelah thawing)

80.0 70.0 60.0 Motilitas


Viabilitas (%)

50.0 (%) 40.0 30.0 20.0 10.0 0.0 SS SE ST

1 jam 2 jam 3 jam

SS Kuda

SE

ST Nama

Nama Kuda

Gambar 3. Persentase motilitas dan viabilitas spermatozoa pada berbagai tahapan pengolahan semen untuk Prince couperos (SS = semen segar; SE = setelah ekuilibrasi; ST = setelah thawing)

80.0 70.0 60.0 Motilitas (%)


Viabilitas (%)

50.0 40.0 30.0 20.0 10.0 0.0 SS SE Nama Kuda ST

1 Jam 2 Jam 3 Jam

SS

SE Nama Kuda

ST

Gambar 4. Persentase motilitas dan viabilitas spermatozoa pada berbagai tahapan pengolahan semen untuk seluruh kuda jantan (SS = semen segar; SE = setelah ekuilibrasi; ST = setelah thawing).

Penentuan Waktu Ekuilibrasi (Arifiantini et al.)

151

45.0 40.0

42.1 39.8 36.9 33.8 30.0 25.1 29.6 28.8 32.2 39.8

42.1 38.1

R ecovery rate (%)

35.0 30.0 25.0 20.0 15.0 10.0 5.0 0.0

1 2 3 Rataan

SJ

SB

PC

Nama kuda

Gambar 5. Recovery rate semen beku kuda dengan waktu ekuilibrasi yang berbeda

Tabel 2. Penurunan kualitas semen pada berbagai tahap pengolahan semen dengan lama ekuilibrasi yang berbeda Nama kuda Spirit of Jatim Waktu Ekuilibrasi M 7,50 6,80 4,90 6,40 6,30 5,00 6,30 5,80 4,40 4,40 6,30 5,00 5,80 SS-SE V 6,20 10,10 11,10 9,10 1,80 2,60 4,70 3,00 4,40 4,40 3,30 4,00 5,50 Penurunan (%) SE-ST M V 35,60 35,30 30,70 24,50 35,10 18,80 33,80 26,20 44,40 35,60 36,90 26,00 36,90 34,90 39,40 32,20 45,60 38,20 38,80 29,60 35,60 25,60 40,00 31,10 37,50 29,70 SS-ST M V 43,10 41,50 37,50 34,60 40,00 29,90 40,20 35,30 50,60 37,40 41,90 28,60 43,10 39,60 45,20 35,20 50,00 42,50 43,10 34,00 41,90 28,90 45,00 35,10 43,30 35,20

1 2 3 Rataan Sony Boy 1 2 3 Rataan Prince 1 Couperous 2 3 Rataan Rataan ke tiga kuda

SS-SE (semen segar ke setelah ekuilibrasi), SE-ST (setelah ekuilibrasi ke setelah thawing), SS-ST ( semen segar ke setelah thawing ), M: motilitas , V, viabilitas spermatozoa

Berdasarkan persentase spermatozoa motil yang dihasilkan setelah thawing dan dibandingkan kualitas semen segar, ternyata nilai RR tertinggi sebesar 42,10% terdapat pada PC (E3 ) dan SB (E2 ), sedangkan nilai RR paling rendah adalah pada SJ (25,10%) dengan waktu ekuilibrasi selama 1 jam. Waktu ekuilibrasi pada semen berbagai ternak berbeda-beda. Pada pembekuan semen domba ekuilibrasi sebaiknya dilakukan selama 2 jam sedangkan pada semen rusa ekuilibrasi selama 4 jam memberikan hasil yang cukup baik (Bearden et al., 2004). Pada semen sapi FH ekuilibrasi biasanya dilakukan selama 4 jam dengan suhu 5 o C. (Arifiantini et al., 2005).

Menurut Raya (2000), ekuilibrasi semen kambing PE pada suhu 3-5 o C selama 4 jam lebih baik dibandingkan dengan 1, 2 ataupun 3 jam. Han et al. (2005) membandingkan lama ekuilibrasi pada pembekuan semen itik yaitu 10 dan 60 menit, ternyata ekuilibrasi selama 10 menit menunjukkan kualitas semen setelah thawing lebih baik dibandingkan dengan 60 menit Untuk semen kuda ekuilibrasi dapat dilakukan selama 1 jam pada suhu 5 o C (Medeiros et al., 2002) ataupun 2 jam (Arruda, et al., 2002). Penurunan kualitas semen setelah ekuilibrasi dan setelah thawing dalam proses pembekuan semen dimulai saat penambahan krioprotektan, perubahan

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volume sel pada saat pembekuan diikuti peregangan dan pengkerutan membran plasma sebagai respon terhadap larutan hiperosmotik (Devireddy et al., 2002) ataupun 2 jam Arruda et al. (2002). Dari hasil penelitian yang telah dilakukan, terlihat bahwa penurunan kualitas semen setelah ekuilibrasi tidak terlalu besar. Tetapi penurunan sangat besar terjadi setelah thawing. Penurunan motilitas spermatozoa dari semen segar sampai setelah thawing pada penelitian ini berkisar antara 37,50% dan 50,60%. Penurunan ini sangat besar, karena menurut Kavak et al. (2003) pada kuda jenis Tori dan Estonian penurunan motilitas dari semen segar ke setelah thawing hanya berkisar antara 24.4 dan 25,20%, demikian juga pada kuda jenis warmblood penurunan motilitasnya sekitar 26% ( Aurich et al., 1996) Penurunan motilitas dan viabilitas spermatozoa setelah thawing ini bervariasi antar individu, hal ini berhubungan dengan permeabilitas dari membran plasma spermatozoa dan efek toksik gliserol yang ditambahkan (Chenier et al., 1998). Menurut White (1993) kerusakan yang terjadi selama proses pembekuan dan thawing karena terbentuknya peroksidasi lipid sehingga akan menurunkan daya hidup spermatozoa. Peroksidasi lipid diawali dengan terbentuknya reactive oxygen species (ROS) sebagai produk samping dari metabolisme spermatozoa. Reactive oxygen species tersebut akan menyerang substrat yang terdekat yaitu ikatan rangkap rantai asam lemak tidak jenuh yang terdapat pada bagian midpiece. Pengaruhnya secara langsung masih dalam penelitian tetapi kelihatannya berhubungan dengan proses kapasitasi sehingga spermatozoa kehilangan fertilitasnya (Gadella et al., 2002). Spermatozoa kuda sangat mudah terkena cold shock. Rentannya spermatozoa kuda ini disebabkan oleh perbedaan komposisi phospholipid yang terdapat pada membran plasmanya. Pada spermatozoa kuda kandungan asam lemak tak jenuh arakidonat (20:4) sangat tinggi yaitu 18,20% (Chow et al., 1986) sedangkan pada spermatozoa sapi dan domba hanya 3,5 dan 4,50-5% (White, 1993). Selain itu perbandingan kandungan antara asam dokosapentaenoat (DPA; 22:5) dan dokosaheksaenoat (DHA; 22:6) pada phosphatidylcholine dan phosphathidyletholamin jumlahnya terbalik (Gadella et al., 2001). Pada spermatozoa sapi dan domba kandungan DPA sangat sedikit, sedangkan pada kuda 17,20%, sebaliknya kandungan DHA pada spermatozoa sapi dan domba sangat tinggi yaitu

61,30 dan 61,40% (White, 1993) tetapi pada spermatozoa kuda hanya 7,60% (Chow et al., 1986). Penambahan gliserol sebagai krioprotektan ke dalam bahan pengencer pada waktu ekuilibrasi membutuhkan waktu yang optimum. Pemaparan gliserol yang terlalu lama atau terlalu singkat sebelum dibekukan akan merusak spermatozoa. Ekuilibrasi yang terlalu lama akan menyebabkan kontak gliserol dengan spermatozoa yang berlebihan sehingga gliserol akan bersifat toksik terhadap sel spermatozoa (sitotoksik). Gliserol akan menarik air secara berlebihan dari dalam sel spermatozoa yang menyebabkan dehidrasi dan selanjutnya terjadi kerusakan sel spermatozoa (Best, 2006). Pada waktu ekuilibrasi gliserol akan memasuki sel-sel spermatozoa untuk menggantikan dari sebagian air yang ada di dalam sel, tetapi dengan waktu ekuilibrasi yang lebih lama kemungkinan gliserol dapat bersifat toksik sehingga akan merusak membran plasma spermatozoa dan menyebabkan gangguan motilitas maupun viabilitas dari spermatozoa (Toelihere, 1993). Dengan waktu ekuilibrasi yang singkat efek sitotoksik gliserol dapat diminimalkan, sehingga penurunan kualitas juga dapat ditekan. Han (2005) menyatakan bahwa efek toksik krioprotektan berhubungan dengan konsentrasi krioprotektan yang ditambahkan dan waktu pemaparan krioprotektan dengan semen sebelum dibekukan. Namun hal tersebut menyebabkan berkurangnya kesempatan gliserol untuk penetrasi secara sempurna ke dalam sel spermatozoa, sehingga terjadi kerusakan spermatozoa karena terbentuknya kristal es intraseluler.

Kesimpulan
Ekuilibrasi selama 1, 2 dan 3 jam tidak menunjukkan perbedaan motilitas spermatozoa setelah thawing pada ketiga kuda yang digunakan. Ekuilibrasi selama 2 dan 3 jam menunjukkan persentase viabilitas spermatozoa lebih baik pada ketiga kuda yang digunakan. Penurunan persentase motilitas tertinggi terdapat pada Sony Boy dengan waktu ekuilibrasi selama 1 jam dan terendah pada Spirit of Jatim selama 2 jam. Penurunan persentase viabilitas spermatozoa tertinggi pada Prince Couperous dengan lama E1 dan terendah pada Sony Boy dengan ekuilibrasi selama 2 jam. Recovery rate terbaik terdapat pada kuda Sony Boy dan Prince Couperous.

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Arifiantini, I., T.L. Yusuf dan N. Graha, 2005. Longivitas dan recovery rate pasca thawing semen beku sapi Fresian Holstein menggunakan bahan pengencer yang berbeda. Buletin Peternakan 29(2): 53-61. Arifiantini, I., T.L. Yusuf dan B. Purwantara, 2006. Daya tahan spermatozoa kuda hasil sentrifugasi dengan kadar plasma semen yang berbeda menggunakan pengencer skim. Journal Animal Production 8(3): 160-167. Arruda, S.P., B.A. Ball, C.G. Gravance, A.R. Garcia and I.K.M. Liu, 2002. Effects of extenders and cryoprotectants on stallion sperm head morphometry. Theriogenology 58:253-256. Aurich J.E., A. Khne, H. Hoppe, and C. Aurich, 1996. Seminal plasma effects membrane integrity and motility of equine spermatozoa after cryopreservation. Theriogenology 46:791-797. Bearden H.J., J.W. Furguay, and S.T. Willard, 2004. Applied Animal Reproduction. 6 th Ed., Pearson Education, New Jersey. Brinsko S.P., G.S. Van Wagner, J.K. Graham and E.L. Quires, 2000. Motility, morphology and triple stain analysis of fresh, cooled and frozen-thawed stallion spermatozoa. Journal of Reproduction and Fertility 56: 111-120 (Supplement). Best, B., 2006. Viability, Cryoprotectant Toxicity and Chilling Injury in Cryonics. www.benbest.com /resources/cryopreservation.pdf. [29 September 2006]. Chenier, T, K. Merkies, S. Leibo, C. Plante, and W. Johnson, 1998. Evaluation of cryoprotective agents for use in the criopreservation of equine spermatozoa. Proceeding of American. Association Equine Practitioners Vol 4. University of Guelph, Guelph, Ontario, Canada. Chow, P.Y.W., I.G. White and B.W. Pickett, 1986. Stallion sperm and seminal plasma phospholipids and glycerylphosphorylcholine. Animal Reproduction Science 11:207-213. Devireddy, R.V., D.J. Swanlund, T. Olin, W. Vincente, M.H.T. Troedsson, J.C. Bischof and K.P. Roberts, 2002. Cryopreservation of equine sperm: Optimal cooling rates in the presence and absence of cryoprotective agents determined using differential scanning calorimetry. Biology of Reproduction 66: 222-231. Dowsett, K,F and L.M. Knott, 1996. The influence of age and breed on stallion semen. Theriogenology 46: 379-412.

Gadella, B.M, R. Rathi, J.F.H.M. Brouwers, T.A.E. Stout, and B. Colenbrander, 2001. Capacitation and the acrosome reaction in equine sperm. Animal Reproduction Science 68:249-265. Gadella, B.M, R. Rathi, M.M. Bevers, J.F.H.M. Brouwers, D. Neild, and B. Colenbrander, 2002. The role of lipid dynamics in equine sperm plasma membrane function. Proceedings of the Second Meeting of the European Equine Gamete Group (EEGG). 26 th 29 th September 2001. Loosdrecht R &W Publications. Han, X.F, Z. Y. Niu, F.Z. Liu and C.S. Yang, 2005. Effect of diluents, cryoprotectants, equilibration time and thawing temperature on cryopreservation of duck semen. Poultry Science 4:197-201. Kacker, R.N., and B.S. Panwar, 1996. Textbook of Equine Husbandry. 1 st Ed., Vikas Publishing House, UK. Kavak, A., A. Johanisson, N. Lundeheim, H. RodriguezMartinez., M. Aidnik and S. Einarsson, 2003. Evaluation of cryopreserved stallion semen from Tory and Estonian breeds using CASA and flow cytometry. Animal Reproduction Science 76: 205216 Kenney, R.M., R.V. Bergman, W.L. Cooper, and G.W. Morse, 1975. Minimal contamination techniques for breeding mares: Technique and preliminary findings. Proceeding of American Association Equine Practitioners:327-336. Medeiros, A.S.L., G.M. Gomes, M.T. Carmo, F.O. Papa, M.A. Alvarenga, 2002. Cryopreservation of stallion sperm using different amides. Theriogenology 58:273-276. Mickelsen, W.D. M.A. Memon, P.B. Anderson, and D.A. Freeman, 1993. The relationship of semen quality to pregnancy rate and litter size following artificial insemination in the bitch. Theriogenology 39: 553560. MacPherson, M.L., 2001. How to evaluate semen in the field. Proceeding of American. Association Equine Practitioners:412-416. Quintero-Moreno, A., J. Miro, J. Teresa-Rigau and J.E. Rodriguez-Gil, 2003. Identification of sperm subpopulation with specific motility characteristics in stallion ejaculates. Theriogenology 59: 19731990. Raya, Y., 2000. Pengaruh waktu ekuilibrasi terhadap kualitas semen beku kambing Peranakan Etawah [Skripsi]. Fakultas Kedokteran Hewan, Institut Pertanian Bogor.

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Steel, R.G.D. dan J.H. Torrie, 1995. Prinsip dan Prosedur Statistika. B Sumantri (penerjemah). Terjemahan dari: Principles and Procedures of Statistics. Gramedia, Jakarta. Toelihere, M.R., 1993. Inseminasi Buatan pada Ternak. Angkasa, Bandung.

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ANIMAL PRODUCTION, September 2007, hlm. 153 159 ISSN 1411 2027 Terakreditasi No.56/DIKTI/Kep/2005

Vol. 9 No.3

Chemical Composition of Leaf and Stem of Tropical Grasses at Different Stages of Growth
(Komposisi Kimia Daun dan Batang Rumput Tropis pada Fase Pertumbuhan yang Berbeda)
Prapti Mahyuddin *
Indonesian Research Institute for Animal Production, Ciawi, P.O.Box 221 Bogor

ABSTRAK: Komposisi kimia daun dan batang dari rumput tropis, Sorghum stipodeum, Themeda australis, Iseilema vaginiflorum, Brachyacne convergens, Dicanthium fecundum (berdaun lebar, bl dan berdaun kecil, nl), Enneapogon polyphyllus, Panicum sp, and Eriachne obtuse dianalisa. Rumput-rumput tersebut mulai dipanen pada saat permulaan berbunga sampai tanaman menjadi kering dengan interval 9, 12, 16, 23 dan 36 hari. Pada semua jenis rumput yang dianalisa, kandungan konstituen dinding sel yang diukur dengan analisis neutral detergent fiber (NDF) lebih tinggi didalam batang dari pada didalam daun (84% vs 75%). Lima dan 8 dari 9 species tersebut berbeda nyata antara batang dan daun masing-masing untuk kandungan ADF (54,20% vs 51,10%) dan lignin (8,60% vs 5,40%). Pada 5 species yang dianalisis, semuanya mengandung protein kasar (CP) lebih tinggi didalam daun dari pada didalam batang (5% vs 2,5%), dan semuanya mengandung karbohidrat larut dalam air (WSC) yang tidak berbeda nyata (3%) didalam daun maupun didalam batang. Tiga dari 5 species yang dianalisis menunjukkan kandungan extrak larut dalam air (WSE) yang lebih tinggi didalam daun dari pada di dalam batang (12,30% vs 9,40%). Enam dari 9 jenis rumput tersebut, mengandung NDF, ADF dan lignin yang tidak berbeda nyata antara periode panen karena rumput-rumput tersebut sudah pada tingkat pertumbuhan yang lanjut. Empat dari 5 species yang dialanisis untuk CP dan 2 dari 5 species yang dianalisa untuk WSE dan WSC menunjukkan kecenderungan penurunan dengan bertambahnya umur tanaman. Diperkirakan bahwa dengan tingginya kandungan NDF dan rendahnya kandungan CP pada rumput-rumput tersebut, akan membatasi konsumsi ternak dan ternak akan kehilangan bobot badan. Daun dapat dijadikan faktor untuk seleksi pada rumput tropis. Kata Kunci: Rumput tropis, batang, daun, dinding sel, pertumbuhan tanaman

Introduction
Reid (1994) defined forage quality as the product of voluntary intake, digestibility and the efficiency of nutrient used by the animal. These parameters are associated with plant anatomichistological characteristics and its chemical composition, which varies with species, cultivar, climate, soil, crop and storage conditions but mainly, stage of plant growth (Buxton and Fales, 2004; Nelson and Moser, 1994; Rotz and Muck, 1994; Van Soest, 1996; Wilson and Hatfield, 1997). When ruminant production system is based on forage as protein and energy sources, the efficiency of nutrient used will be highly dependent on forage maturity, which is often considered to be the primary factor that determines nutritional quality.

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There is a general agreement that low quality is inherent characteristic of tropical grasses, mainly as a result of a high content of cell wall constituent (CWC). This places increased emphasis on the microbial and mechanical processes of digestion in the reticulorumen, especially with respect to limitation of voluntary intake. The high content of CWC also places increased importance on the digestion of CWC since it represents a high proportion of the total digestible organic matter. Lignin contained in CWC is insoluble, unaffected by most enzymes, and acting as a constraint on polysaccharide utilization by forming an encrusting barrier. As plant mature, the ratio of leaf: stem usually declines, resulting in reduced forage quality as lignin content in developing stems increases and protein content and digestibility decline (Colleman, 1992; Van Soest, 1996). This paper reports the chemical composition of leaf and stem at different stage of growth in some tropical grasses.

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Research Methods
Treatments
There were nine species of grasses used in this experiment: Sorghum stipodeum, Themeda australis, Iseilema vaginiflorum, Brachyacne convergens, Dicanthium fecundum (broad and narrow leaf), Enneapogon polyphyllus, Panicum sp, and Eriachne obtuse. The forage grew under natural conditions, therefore the exact day of germination of any plant sampled was not known. However, the stages of growth were recorded at each time they were harvested. Harvesting was carried out at interval of 9, 12, 16, 23 and 36 days. The stage of maturity of the grasses at each time of harvesting is presented in Table 1. All grasses were harvested at the ground level. Unfortunately, neither the production nor the leaf/stem ratios were possible to be determined. Samples of grasses were separated into their botanical component, leaf and stem. Since the separation is time-consuming, being done by hand, the petiole and the leaf were not separated but mixed together as one leaf sample. Prior to grinding, they were oven dried at 80 0C overnight. Samples were ground to pass through 1 mm sieve in a small laboratory mill. After being ground, they kept in closed containers, ready for chemical analysis. All analysis was determined on a DM basis (constant weight). Duplicate samples were included in each run and repeated if the difference between the duplicate exceeded 2 percentage units. The water soluble extract (WSE), water soluble carbohydrate (WSC), and crude protein were determined only on the 5 species. Cell wall constituent (CWC) were described by detergent fiber analysis (Goering and Van Soest, 1970). Neutral detergent fiber (NDF) was assayed to define the amount of total CWC present in the grass sample. Acid detergent fiber (ADF) was determined, and residues were used in analyses for cellulose, lignin and acid insoluble ash content (Goering and Van Soest, 1970). ADF contains cellulose, lignin and acid insoluble ash. ADF residue was reacted with 72% H 2SO 4. It is assumed that all cellulose was dissolved in 72% H 2 SO 4. The residue was assumed to be lignin and acid insoluble ash. Lignin was calculated as weight loss after ashing the residue. Percent crude protein (N x 6.25) was calculated from the amount of nitrogen in samples as determined by micro Kjeldhal method (AOAC, 1995).

Statistical Analysis
Not being a field trial, this experiment has no replication. In order to obtain an estimate of error, we must assume a linear relationship between the response and the stage of maturity. The analysis fitted Part as a qualitative factor, enabling to assess whether leaf and stem have different concentration of the chemical component. We regressed linearity on Period to see if there is a steady change over time. Interaction between Part and Period was also fitted to test for different responses between stem and leaf over stage of maturity. Data on chemical composition were subjected to one-way analysis of variance using the general linear model of SAS (1988). Multiple comparisons was done using Duncans multiplerange test (Snedecor and Cochran, 1980).

Results and Discussion


The CWC measured as NDF content was higher in the stem than in the leaf (84% vs. 75%) of all species studied (Table 2 and Table 8), although the difference in the fiber component (ADF and lignin) was either non significant or the content of the stem was higher than that of the leaf fraction. Except Sorghum, all species showed a significantly higher lignin content in the stem than that in the leaf (8.20% vs. 5.60%); whereas for ADF content, 5 out of 9 species studied showed the value in the stem was higher than that in the leaf (55% vs. 51%) (Table3, Table 4, and Table 8). This result is in agreement with what stated by Colleman (1992) and Van Soest (1996). Three out of 5 species measured showed that WSE content was higher in the leaf than that in the stem (12.30% vs. 9.40%) (Table 6 and Table 8) and none of them had no significant difference between leaf and stem (3%) for WSC (Table 7 and Table 8). However, all species analyzed indicated higher CP content in the leaf than in the stem (5% vs. 2.50%) (Table 5 and Table 8). These results could be due to differences in the composition of the tissue type that constitute the plant fraction, and thus the differences may vary with the species. Akin et al. (1977) studied the histochemistry of 5.5 months Coastal Bermuda Grass, and found that the composition of the tissue types that constitute sheath (leaf) was different from that of the stem fraction, and as a result the chemical composition of these fractions was different also.

Cemichal Composition of Leaf (Mahyuddin)

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When the chemical content of the stem and leaf differed, the difference at each harvest varied with species. This difference can be constant, suggesting that the rate of change in the chemical content over the period of harvesting was similar for both fractions. The difference may also increase as maturity advances, and this suggests that the increase in the chemical content with maturity was faster in the stem than in leaf or vice versa. This may be associated with the accumulated and

translocation of assimilates to or from the stem or leaf. For example, in species where increase in CWC with maturity was faster in stem than in leaf, the stem may translocate more assimilate for seed production than the leaf, this in turn causes higher CWC content in stem than in leaf. When the decrease in soluble material was faster in leaf than in stem, the leaf translocates assimilates, more than in stem.

Table 1. Plant species and stage of maturity at each harvest Harvesting period 1 2 3 4 5 Sorghum Themeda Iseilema Early flowering Late flowering Setting seed Seed shedding Drying off Brahyacne Dicanthium (bl) Dicanthium (nl) Late flowering Setting seed Late seed set Seed shedding Drying off Enneapogon Panicum Eriachne Setting seed Late seed set Seed shedding Seed shedding Drying off

bl = broad leaf, nl = narrow leaf

Table 2. Neutral detergent fiber (% DM) content of stem and leaf at different stage of growth Species Sorghum Themeda Iseilema Brachyacne Dicanthium (bl) Dicanthium (nl) Enneapogon Panicum Eriachne Average SE S 76.50 85.40 82.10 82.50 82.60 82.70 81.80 80.00 86.60 82.20 0.97 EF L 71.30 71.70 74.00 78.20 74.40 70.60 73.00 72.80 81.40 74.10 1.17 S 69.30 83.00 82.40 79.10 84.20 82.50 78.90 79.00 84.00 80.30 1.54 LF L 67.00 73.30 72.30 75.30 74.70 75.30 70.70 69.50 80.10 73.10 1.28 S 88.00 79.20 80.10 79.50 81.60 85.00 80.10 84.10 86.10 82.60 1.08 SS L 75.20 70.40 73.40 76.40 75.30 76.10 71.00 75.60 78.40 74.60 0.88 SSh S L 86.20 76.30 81.30 73.00 82.40 73.70 78.90 75.70 81.00 73.50 81.20 75.00 84.00 73.40 83.00 79.40 86.20 81.40 82.70 75.70 0.81 0.97 DO S L 90.70 76.10 82.40 73.90 85.30 76.50 81.10 77.00 83.50 74.70 86.10 80.50 86.10 72.40 87.60 80.20 85.00 82.50 86.30 77.10 0.95 1.12

DM: Dry matter, EF: Early Flowering, LF: Late Flowering, SS: Setting Seed, SSh: Seed Shedding, DO: Drying Off, S: Stem, L: Leaf, SE: standard error, bl : broad leaf, nl : narrow leaf

Table 3. Acid detergent fiber (% DM) content of stem and leaf at different stage of growth Species Sorghum Themeda Iseilema Brachyacne Dicanthium (bl) Dicanthium (nl) Enneapogon Panicum Eriachne Average SE EL S L 49.60 41.30 53.50 49.10 59.20 58.60 53.80 54.20 52.40 48.20 52.70 48.50 55.40 55.30 49.60 47.40 53.80 58.90 53.30 51.30 0.97 1.94 LF S 39.60 61.30 60.00 54.40 62.60 50.80 47.00 54.50 44.40 52.70 2.65 L 47.20 46.80 57.00 42.00 54.00 46.30 47.00 45.00 50.30 48.40 1.54 SS S 63.70 53.10 56.20 49.60 58.40 57.50 53.30 51.60 50.90 54.90 1.48 L 47.60 46.50 53.50 49.10 56.10 51.70 53.70 47.20 56.70 51.30 1.29 SH S L 57.20 52.10 55.10 50.20 58.00 56.30 50.20 47.80 61.40 52.80 53.90 48.70 54.50 54.00 53.70 50.40 47.50 52.50 54.60 51.60 1.37 0.88 DO S L 62.10 54.70 53.60 49.20 64.00 63.40 50.00 51.70 59.10 54.30 55.40 57.30 58.10 53.30 57.30 53.30 50.90 56.10 56.70 54.80 1.58 1.33

DM: Dry matter, EF: Early Flowering, LF: Late Flowering, SS: Setting Seed, SSh: Seed Shedding, DO: Drying Off, S: Stem, L: Leaf, SE : standard error, bl : broad leaf, nl : narrow leaf

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Table 4. Lignin content (% DM) in stem and leaf at different stage of growth Species Sorghum Themeda Iseilema Brachyacne Dicanthium (bl) Dicanthium (nl) Enneapogon Panicum Eriachne Average SE EL S 4.40 8.40 9.30 11.10 5.60 6.40 9.00 4.90 8.60 7.50 0.76 L 3.60 6.80 6.70 7.10 3.80 5.00 4.70 2.60 6.50 5.00 0.55 LF S 4.80 8.60 10.70 9.20 10.30 6.00 7.60 8.00 7.00 8.00 0.64 L 5.60 5.60 6.10 4.90 5.00 3.70 4.60 4.20 6.50 5.10 0.30 SS S 6.50 7.60 9.60 7.60 7.70 7.10 12.90 6.40 14.30 8.80 0.95 L 5.90 4.80 5.90 5.70 5.80 5.50 3.70 4.80 7.10 5.30 0.31 SH S 7.00 8.60 9.40 7.20 8.90 7.30 8.30 7.20 8.00 8.00 0.29 L 6.40 6.10 5.90 4.50 5.00 4.50 4.40 3.50 5.60 5.10 0.32 DO S 7.60 8.70 11.80 8.60 9.80 8.90 10.60 8.10 8.10 9.10 0.45 L 7.90 6.40 7.00 5.10 5.50 6.80 4.70 4.40 8.50 6.20 0.47

DM: Dry matter, EF: Early Flowering, LF: Late Flowering, SS: Setting Seed, SSh: Seed Shedding, DO: Drying Off, S: Stem, L: Leaf, SE : standard error, bl : broad leaf, nl : narrow leaf

Table 5. Crude protein content (%DM) of leaf and stem at different stage of growth Species Sorghum Themeda Iseilema Brachyacne Dicanthium (bl) Average S 3.50 2.20 2.10 5.10 3.20 3.20 EL L 6.00 5.40 3.30 5.60 6.90 5.40 S 3.30 1.90 1.10 6.10 1.70 2.80 LF L 12.40 4.70 2.90 8.60 4.60 6.60 S 1.80 1.40 1.40 6.20 1.70 2.50 SS L 5.20 4.80 2.80 8.10 5.10 5.20 S 1.70 1.00 1.10 4.20 1.30 1.90 SH L 2.30 2.50 1.80 5.60 3.80 3.20 S 0.80 0.80 0.70 2.70 1.40 1.20 DO L 1.40 3.10 1.40 3.90 3.90 2.70

DM: Dry matter, EF: Early Flowering, LF: Late Flowering, SS: Setting Seed, SSh: Seed Shedding, DO: Drying Off, S: Stem, L: Leaf, bl : broad leaf

Table 6.Water soluble extract (% DM) of stem and leaf at different stage of growth Species Sorghum Themeda Iseilema Brachyacne Dicanthium (bl) Average SE EL S 19.20 6.50 9.90 12.50 9.50 11.50 2.14 L 15.60 12.30 9.40 14.50 15.40 13.40 1.17 LF S 23.90 7.50 9.70 14.90 7.30 12.70 3.13 L 17.20 11.10 10.80 16.50 11.40 13.40 1.42 SS S 7.80 10.10 11.20 12.80 7.50 9.90 1.01 L 11.90 13.50 9.10 14.80 10.30 11.90 1.03 SH S 8.50 7.90 7.90 14.00 8.90 9.40 1.15 L 10.40 10.80 7.40 16.00 11.50 11.20 1.38 DO S 3.90 5.70 5.70 9.20 6.40 6.20 0.86 L 7.80 6.80 4.30 10.60 9.10 7.70 1.06

DM: Dry matter, , EF: Early Flowering, LF: Late Flowering, SS: Setting Seed, SSh: Seed Shedding, DO: Drying Off, S: Stem, L: Leaf, SE : standard error, bl : broad leaf

Table 7. Water soluble carbohydrate (% DM) of stem and leaf at different stage of growth Species Sorghum Themeda Iseilema Brachyacne Dicanthium(bl) Average SE S 7.50 1.00 2.10 1.20 1.60 2.70 1.22 EL L 5.00 3.40 1.30 1.60 1.80 2.60 0.70 S 7.80 2.10 4.30 2.60 3.10 4.00 1.15 LF L 7.00 3.80 2.80 3.30 2.70 3.90 0.80 S 3.60 5.90 4.80 2.10 3.30 3.90 0.65 SS L 3.20 5.00 2.60 3.10 2.30 3.20 0.46 S 4.30 5.60 1.80 3.60 5.80 4.20 0.73 SH L 3.70 4.50 1.70 3.20 3.00 3.20 0.46 S 1.20 3.70 1.20 3.00 3.20 2.50 0.53 DO L 1.60 3.20 0.90 3.40 2.20 2.30 0.47

DM: Dry matter, EF: Early Flowering, LF: Late Flowering, SS: Setting Seed, SSh: Seed Shedding, DO: Drying Off, S: Stem, L: Leaf, SE : standard error, bl : broad leaf

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Table 8. Summary of statistical analysis of chemical component vs. plant part Species Sorghum Themeda Iseilema Brachyacne Dicanthium(bl) Dicanthium(nl) Enneapogon Panicum Eriachne + + + + + + + + + CWC S>L + S>L + S>L + S>L + S>L + S>L + S>L + S>L + S>L ADF NS ++ NS NS + + NS ++ + S>L Lignin NS + + S>L + + S>L + + S>L + + S>L + + S>L + + S>L + + S>L + + S>L + + + + + + + + CP L>S L>S L>S L>S + L>S WSE NS + + L>S NS + + L>S + + L>S WSC NS NS NS NS NS

S>L S>L S>L S>L

+ : significant at 5 % level, + + : significant at 1 % level, NS: non significant, CWC: Cell wall constituent, ADF: Acid detergent fiber, CP: crude protein, WSE: Water soluble extract, WSC: Water soluble carbohydrate, S: Stem, L: Leaf, bl : broad leaf, nl : narrow leaf

Cell Wall Constituent (CWC)


In all of the samples the value of NDF was very high, ranging from 69% to 90% in the stem and from 67% to 84% in the leaf fraction. This is not surprising as all the sample material is already in an advanced stage of growth (the earliest stage being flowering). Therefore in this study, 6 out of nine species showed no difference in NDF and ADF content with period of harvesting (Table 9). In an earlier stage, The CWC of tropical grasses should have been lower, although it may be still higher than that of temperate grasses at the same stage of growth (Minson, 1990) ADF consists mainly of lignin, cellulose and acid insoluble ash (AIA). Since AIA contributes only a very small proportion of the total quantity, cellulose and lignin are the main components which influence the behavior of ADF. Lignin and ADF have been suggested as being useful for prediction of digestibility (Van Soest and Mertens, 1977). As lignin often limits the availability of cellulose, the association of ADF with digestibility depends largely upon the association of lignin with cellulose. The concentration of lignin was always higher in the stem than that in the leaf fraction, except in Sorghum. However, only 3 out of 9 species studied, showed an increase in lignin content with advancing maturity. Lignin which is part of CWC by NDF determination may loss in the cell content (neutral detergent-soluble) appear as lignin-carbohydrate complex (Lowry et al., 2002). Although in this study such lignin-carbohydrate complex was not determined, result from Lowry et al. (2002) showed lignin-carbohydrate complex could account for 3.50%, 3.60%, 4.20% and 7.90% in cell content of Iseilema, Panicum, Sorghum and Themeda

respectively. It may be anticipated that for those species, the correlation between lignin (measured through ADF determination) and digestibility would be low.

Soluble Materials
While structural carbohydrate, the CWC increased with maturity, the concentration of water soluble extract (contains some WSC and some protein) usually decreased with advancing maturity. Like CWC, however, the response to maturity depends upon the species. Of the five species measured, WSE decreased in Sorghum and Iseilema, WSC decreased in Sorghum and increased in Themeda, whereas crude protein decreased in all species except in Dicanthium. The content of WSC in forage is a useful guide to silage making. While the mean value of WSC in this study was 3%, Yang et al. (2006) found that the grass initial WSC content of at least 7% is required for the lactic acid bacteria to multiply to drop the pH to 4 (the best pH silage is 3.8 4.2). The low WSC content in this study may be due to the samples were oven dried, it would be higher if they were analyzed as fresh sample (Piccaglia and Galetti, 1987; Kerepesi et al., 1996).

Application
The high CWC concentration (>60%) in these tropical grasses may have an effect on voluntary intake. The limitation of intake caused by the high CWC content can be expected particularly when it is lignified for it may limit the accessibility of microbial enzymes to substrate (Wilson and Hatfield, 1997) and therefore may prolong the residence time of feed material in the reticulorumen.

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Table 9. Summary of statistical analysis of chemical component vs. period Species CWC ADF Lignin CP Sorghum + + ++ + Themeda NS NS NS ++ Iseilema + NS NS ++ Brachyacne NS NS NS ++ Dicanthium (bl) NS NS NS NS Dicanthium (nl) NS ++ ++ Enneapogon NS NS ++ Panicum ++ + NS Eriachne NS NS NS

WSE ++ NS ++ NS NS

WSC ++ ++ NS NS NS

+ : significant at 5 % level, + + : significant at 1 % level, NS: non significant, CWC: Cell wall constituent, ADF: Acid detergent fiber, CP: crude protein, WSE: Water soluble extract, WSC: Water soluble carbohydrate, S: Stem, L: Leaf, bl : broad leaf, nl : narrow leaf

Considering that the crude protein (CP) content of the plant material was less than 7% which is below the dietary requirement of maximum bacterial growth (11% to 14%) (Satter and Roffler, 1975), it would be expected that animals may loose weight for stored energy would be used. Minson (1990), reported that 7% CP is the level at which nitrogen begin to limit intake in sheep. Feed intake may be limited by the nitrogen status of the animal. And feed intake may increase as the nitrogen status is increased. However, since dietary protein is often degraded by micro-organism in the rumen, a dietary supplement may not be efficient for satisfying the protein requirement unless it is protected from digestion in the rumen. The amount of protein that the animal has available for utilization is dependent upon the undegraded feed protein and microbial protein flowing from the rumen to the small intestine. In condition where the protein requirement is not met, the inclusion of NPN such as urea (for microbial growth) and protected protein in the diet may be useful. However, this may be an expensive solution. Inclusion of forage legume in the swards could offer several advantages to farming system. This cover crop is able to conserve soil, improve organic matter content and compete with weeds (Humpreys, 1995), convert atmospheric nitrogen (N) to form plant N and cycle within the plant-animal-soil system. The legume-rhizobial symbiosis provides farmers with an inexpensive source of N while production is environmentally clean. This symbiosis does not involve the consumption of soil fuel as occurs in the production of fertilizer N which contributes to global warming. N fixing legumes have higher concentration of cellular protein than tropical grasses as such. However, the different growing habits and optimum harvesting frequencies between

grasses and legumes makes them difficult to maintain in the mixed sward. Following regular harvesting, the grass would quickly shade legumes, thus reducing their growth rates. Moran (2005) suggested, to maintain the longevity of the forage legumes, they should be sown as a single species sward. Tropical legumes are rich in protein which is usually the most limiting nutrient in tropical animal diets. Tree legumes offer another choice to improve tropical grasses. Usually they are planted in an alley or rows and can be easily harvested at different intervals than the grass. The photosynthetic efficiency of the grass can be improved when the grass grown in association with tree legumes. During dry season the tree will protect the grass from drought. In addition to that some of the protein (tannin-protein complex) in tropical tree legumes are protected from degradation in the rumen (McSweeney et al., 1999) and thus may be better absorbed by the animal. As maturity progresses the protein content decreases and the CWC increases in concentration, and therefore it is advisable to harvest at an immature stage (Moran, 2005). In evaluating forage quality, chemical composition alone cannot be used for predicting forage quality. Concentration perse is not the only attribute to a chemical factor depressing or influencing digestibility; solubility or insolubility of the masking substance and its distribution over potentially digestible substrate surface is also important. Leafiness seems to be another factor that should be considered when evaluating forage quality. As indicated in the present results, leaf tends to have a lower cell wall constituent and a higher soluble material than those in stem. Thus, the ratio of leaf/stem is an important factor affecting diet

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selection and quality of a forage. It has been recently found that near-infra red reflectance spectroscopy can be used as a rapid means of estimating leaf/stem ratio of grasses (Smart et al., 2004).

Moran, J., 2005. Growing quality forages. In: Tropical dairy farming: feeding management for small holder dairy farmers in the humid tropics. Landlinks Press. Pp.312. Nelson, C.J. and L.E. Moser, 1994. Plant factors affecting forage quality. In: Forage quality, Evaluation, and Utilization. G.C. Fahey Jr. (Ed), ASA-CSSA-SSSA, Madison, Pp 115-154. Piccaglia, R and G.C. Galetti, 1987. Effect of sample drying on the determination of fiber and watersoluble carbohydrate of maize silage. Grass and Forage Science 42 (3): 319 321. Reid, R.L., 1994. Milestones in forage research (19691994). In: Forage Quality, Evaluation, and Utilization. G.C Fahey Jr, (Ed.), ASA-CSSA-SSSA, Madison. Pp. 1 -58. Rotz, C.A. and R.E. Muck, 1994. Changes in forage quality during harvest and storage. In: Forage Quality, Evaluation, and Utilization. G.C. Fahey Jr. (Ed.). ASA, CSSA, SSSA, Madison. Pp. 828 868. SAS Institute Inc. 1988. SAS/STAT Users Guide. SAS Institute Inc., Cary, NC, USA. Satter, L.D. and R.E. Roffler, 1975. Nitrogen requirement and utilization in dairy cattle. Journal of Dairy Science 58: 1219- 1237. Smart, A.J, W.H. Schach, L.E. Moser and J.D. Volesky, 2004. Prediction of leaf/stem ratio using nearinfrared reflectance spectroscopy (NIRS). Agronomy Journal 96: 316 318. Snedecor, G.W and W.G. Cochran, 1980. Statistical Methods. 7 th Edision. The Iowa State University Press, Ames. Van Soest, P.J. and D.R. Mertens, 1977. Analytical parameters as guide to forage quality. In: Proceeding of International Meeting of Animal Production, Temperate Grassland. Dublin, Ireland. Pp. 50 52. Van Soest, P.J., 1996. Environment and forage quality. In: Proceedings of Cornell Nutrition Conference for Food Manufacturers, Cornell University, New York. Pp. 1 9. Wilson, J.R. and R.D. Hatfield, 1997. Structural and chemical changes of cell wall types during stem development: consequences for fiber degradation by rumen microflora. Australian Journal Agriculture Research 48(2): 165 180. Yang, H.Y., X. F. Wang, J. B. Liu, L. J. Gao, M. Ishii, Y. Igarashi and Z.J. Cui, 2006. Effects of water-soluble carbohydrate content on silage fermentation of wheat straw. Journal of Bioscience and Bioengineering 101(3): 232 237.

Conclusion
Mature tropical grasses contain very high cell wall constituents and very low crude protein. The leaf has lower cell wall constituents than the stem. It is recommended to harvest at immature stage and select grasses with more leaves than the stems.

References
Akin, D.E., E.L. Robinson, F.E. Barton and D.S. Himmelsback, 1977. Changes with maturity in anatomy, histochemistry, chemistry and tissue digestibility of Bermuda grass plant parts. Journal of Agriculture and Food Chemistry 25: 179. [AOAC] Association of Official Analytical Chemistry, 1995. Official method of analysis. 17 th Ed. Association of Official Analytical Chemistry. Washington DC. Buxton, D.R., and S.L. Fales, 2004. Plant environment and quality. In: Forage Quality, Evaluation, and Utilization. G.C. Fahey Jr., (Ed.). ASA-CSSA-SSSA, Madison, Pp. 155 199. Colleman, S. W., 1992. Plant-animal interface. Journal of Production and Agriculture 5: 7 13. Goering, H.K., and P.J. Van Soest, 1970. Forage fiber analyses (apparatus, reagent, procedures and some applications). USDA Agriculture Handbook. Humphreys, L.R., 1995. Diversity of Productivity of Tropical Legumes. In: Tropical Legumes in Animal Nutrition; DMello, J. P. F and C. Devendra (Eds.). CAB International, Wallingford, Pp. 1-21. Kerepesi, L., M. Toth and L. Boross, 1996. Water-soluble carbohydrate in dried plant. Journal of Agriculture and Food Chemistry 44(10): 3225 3239. Lowry, J.B, P.M. Kennedy and L.L Conlan, 2002. Lignin in the cell content fraction of tropical forages. Journal Science Food Agriculture 83: 370 -374. McSweeney, C.S, B.Palmer, R. Bunch and D.O. Krause, 1999. Isolation and characterization of proteolytic ruminal bacteria from sheep and goats fed the tannincontaining shrub legume Calliandra calotyrsus. Applied and Environmental Microbiology 65(7): 3075 3083. Minson, D.J., 1990. Forage in Ruminant Nutrition. Academic Press, Toronto.

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Vol. 9 No.3

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Vol. 9 No.3

Komposisi Kimia dan Degradasi Nutrien Silase Rumput Gajah yang Diensilase dengan Residu Daun Teh Hitam
(Chemical Compositions and Nutrient Degradation of Elephant Grass Silage Ensiled with Black Tea Waste)
Budi Santoso*, Marlyn Nelce Lekitoo dan Umiyati
Jurusan Nutrisi dan Makanan Ternak, Universitas Negeri Papua, Manokwari

ABSTRACT: This study evaluated the chemical compositions and nutrient degradation during ensiling of elephant grass (Pennisetum purpureum) silage with black tea waste (BTW) addition. Four silage treatments were elephant grass (S0); elephant grass + 100 g BTW/ kg fresh matter (S1); elephant grass + 200 g BTW/kg fresh matter (S2 ); elephant grass + 300 g BTW/kg fresh matter. About 220 g of silage material were ensiled for 30 days at room temperature (approximately 28C). Three replicates were prepared for each treatment. Results showed that dry matter, organic matter and crude protein contents of silages increased linearly (P<0.01) with increasing black tea waste. There were linear decreases in dry matter and crude protein degradations (P<0.01) and organic matter degradation (P<0.05) during ensiling with increased black tea waste addition. Dry matter degradation values varied from 15.03 to 30.71% and were higher than degradation value of ideal silage. It was concluded that black tea waste has potential as a silage additive to improve nutritive value and fermentation quality of elephant grass silage. Key Words: Elephant grass, black tea waste, silage additive, degradation

Pendahuluan
Selama ensilase, sebagian protein dari rumput mengalami degradasi (proteolisis) baik oleh enzim protease tanaman maupun mikroba menjadi senyawa NPN (non-protein nitrogen) terutama asam amino dan amonia (Ohshima dan McDonald, 1978). Untuk mempertahankan dan meningkatkan kandungan nutrisi silase maka dapat dilakukan penambahan aditif seperti kultur bakteri (bakteri asam laktat), sumber karbohidrat mudah larut dalam air, asam organik, enzim, dan nutrien (urea, amonia, mineral mineral) (McDonald, 1981). Formaldehid dapat digunakan sebagai aditif silase untuk meningkatkan kualitas fermentasi karena dapat memproteksi protein dari hidrolisis oleh enzim tanaman dan mikroba, sedangkan asam format dapat menurunkan pH silase sehingga menghambat aktifitas protease dari tanaman (Henderson, 1993). McDonald (1981) menyatakan bahwa penurunan pH dapat pula mencegah pertumbuhan mikroba yang tidak diinginkan seperti Listeria, Clostridia dan jamur.

Penulis Korespondensi. email: budi_santoso@unipa.ac.id

Namun demikian kedua bahan kimia tersebut bersifat toksik yang dapat menyebabkan gangguan pada kulit, mata serta iritasi pernapasan sehingga dapat membahayakan ternak. Menurut Chamberlain (1987) dan Henderson (1993) beberapa kelebihan atau keunggulan penggunaan bahan aditif yang bersifat biologi dibandingkan aditif bersifat kimiawi yaitu aman untuk ditangani, tidak bersifat toksik terhadap ternak, tidak bersifat korosif serta aman dan ramah terhadap lingkungan. Tanin merupakan senyawa sekunder yang terdapat pada tumbuhan dan dapat bereaksi dengan protein, sehingga menyebabkan protein resisten terhadap degradasi oleh protease di dalam silo dan di dalam rumen. Menurut Salawu et al. (1999) dan Tabacco et al. (2006), penambahan bahan sumber tanin yang berasal dari tanaman chestnut atau mimosa dapat meningkatkan kualitas fermentasi silase, ditandai dengan penurunan degradasi bahan kering dan protein kasar selama ensilasi, serta konsentrasi N-amonia/total N. Menurut Kondo et al. (2004b), daun teh hijau mengandung protein, vitamin, tanin dan polifenol terutama katekin, gallat epikatekin, dan gallat epigallokatekin. Sebagian nutrien tersebut masih tersisa walaupun daun teh telah diekstrak dengan menggunakan air panas. Kondo et al. (2004a) melaporkan bahwa penambahan residu daun teh

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hijau pada bahan silase yang berasal dari limbah tanaman gandum meningkatkan kandungan protein kasar. Kualitas fermentasi silase yang ditambahkan residu daun teh hijau lebih baik, ditandai dengan pH<4,13, konsentrasi amonia/total N rendah dan konsentrasi asam laktat lebih tinggi dibandingkan dengan silase kontrol. Disamping itu, silase yang ditambahkan residu daun teh hijau mengeluarkan aroma daun teh sehingga meningkatkan preferensi ternak terhadap silase. Pada percobaan lain Kondo et al. (2004b) melaporkan bahwa kandungan N dan tanin meningkat dua kali lebih tinggi pada silase rumput sudan yang ditambahkan residu daun teh hijau dan teh hitam sebanyak 200 g/kg bahan segar silase dibandingkan silase kontrol. Indonesia merupakan salah satu negara produsen dan eksportir teh dunia dan menempati urutan kelima setelah Sri Langka, Kenya, China dan India. Sekitar 98% teh yang beredar di pasaran adalah teh hitam yang dihasilkan dari fermentasi daun teh hijau. Bubuk teh hitam yang telah diseduh dan disaring menghasilkan ampas (residu) daun teh dan kemudian dibuang (Purnomo, 2004), sehingga dapat digunakan sebagai aditif silase. Penelitian ini bertujuan mengetahui komposisi kimia, yaitu bahan kering (BK), bahan organik (BO), dan protein kasar (PK), dan nilai degradasi nutrien (BK, BO dan PK) dalam silo dari silase rumput gajah (Pennisetum purpureum) yang diensilasi dengan residu daun teh hitam.

sebanyak 850g. Masing-masing bagian ditambahkan aditif residu daun teh hitam dengan konsentrasi sebagai berikut : S0 : rumput gajah saja (kontrol) S1 : rumput gajah + residu daun teh hitam 100 g/kg bahan segar S2 : rumput gajah + residu daun teh hitam 200 g/kg bahan segar S3 : rumput gajah + residu daun teh hitam 300 g/kg bahan segar Masing-masing bahan silase yang telah tercampur homogen dimasukkan ke dalam botol fermentor berkapasitas 220 g dengan 3 replikasi. Bahan silase dipadatkan, kemudian ditutup rapat dan ditimbang beratnya. Botol disimpan pada ruang bertemperatur 28C selama 30 hari. Setelah masa penyimpanan tersebut, botol ditimbang kemudian sampel silase dikeringkan dalam oven 60C selama 48 jam. Selanjutnya sampel digiling menggunakan Wiley mill yang dilengkapi dengan saringan berukuran 1 mm, dan digunakan dalam analisis. Nilai degradasi BK, BO dan PK selama ensilase dinyatakan dalam (%) dan dihitung berdasarkan selisih antara nutrien (BK, BO dan PK) bahan silase dengan nutrien (BK, BO dan PK) setelah menjadi silase, sebagaimana dikemukakan Yahaya et al. (2002).

Analisis Sampel
Kandungan BK, BO, dan PK pada sampel rumput gajah, residu daun teh hitam dan silase dianalisis berdasarkan prosedur yang dikemukakan oleh Harris (1970). Konsentrasi total tanin dianalisis menggunakan metode Folin-Ciocalteu, sebagaimana dideskripsi oleh Santoso et al. (2006).

Metode Penelitian
Penyediaan dan Preparasi Residu Daun Teh Hitam
Daun teh yang digunakan pada penelitian ini merupakan residu daun teh hitam (Cap Rejeki Prima) yang telah diseduh. Daun teh dikering anginkan selama 1 minggu, kemudian digunakan sebagai aditif dalam pembuatan silase. Penyediaan Rumput Gajah dan Preparasi Silase Rumput gajah dipotong dari Laboratorium Lapangan, Fakultas Peternakan Perikanan dan Ilmu Kelautan, Universitas Negeri Papua pada umur 60 hari (Agustus Oktober 2006) setelah pemotongan awal (penyeragaman). Rumput dilayukan pada temperatur ruangan (28C) selama 48 jam kemudian dicacah dengan ukuran 1 2 cm. Rumput yang telah dicacah dicampur hingga homogen kemudian dibagi menjadi 4 bagian, masing-masing

Analisis Statistik
Data dianalisis dengan analisis ragam menurut rancangan acak lengkap menggunakan program SAS versi 6,12 (SAS, 1996). Untuk mengetahui pengaruh konsentrasi residu daun teh (linier atau kuadratik) digunakan uji polinomial ortogonal.

Hasil dan Pembahasan


Komposisi Kimia Rumput Gajah dan Residu Daun Teh Hitam
Rumput gajah yang digunakan pada penelitian ini mengandung BK 22,39%, dan lebih rendah dari BK (25 35%) yang dibutuhkan untuk pembuatan silase yang baik (Chamberlain and Wilkinson,

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1998). Sementara itu, kandungan PK rumput gajah pada penelitian ini lebih tinggi dari pada standar kebutuhan PK minimal untuk ternak ruminansia sebesar 7%, sebagaimana direkomendasikan oleh Crowder and Chheda (1982). Komposisi kimia rumput gajah dan residu daun teh hitam yang digunakan pada penelitian ini secara lengkap disajikan pada Tabel 1.
Tabel 1. Komposisi kimia rumput gajah dan residu daun teh hitam Komposisi kimia BK (%) BO (%) PK (%) Total Tanin (%) Rumput Gajah 19,94 88,83 12,23 Residu Daun Teh Hitam 55,48 95,72 18,12 0,24

BK: bahan kering, BO: bahan organik, PK: protein kasar

Menurut Hartadi et al. (1993), rumput gajah dengan umur potong 57 - 70 hari mengandung PK sebesar 8,30%, sedangkan pada penelitian Sudirman et al. (2006) dilaporkan bahwa rumput gajah mengandung PK sebesar 7,80% Perbedaan nilainilai tersebut dapat disebabkan perbedaan lokasi tempat penanaman yang berhubungan dengan ketersediaan N di dalam tanah. Kandungan BK residu daun teh hitam yang digunakan dalam penelitian ini sama dengan residu daun teh hitam yang dilaporkan oleh Kondo et al. (2004b). Namun demikian kandungan BO, PK dan total tanin pada residu daun teh hitam relatif lebih rendah berturutturut 95,72 vs. 96,90%, 18,12 vs. 24,69% dan 0,24 vs. 0,70%. Komposisi kimia silase rumput gajah yang diensilase dengan residu daun teh hitam disajikan pada Tabel 2. Penambahan residu daun teh hitam meningkatkan kandungan BK silase secara linier (P<0,01). Kandungan BK silase rumput gajah yang ditambahkan residu daun teh hitam sebanyak 100 g, 200 g dan 300 g/kg rumput segar meningkat sebesar 26,62, 56,04 dan 82,04% dibanding perlakuan kontrol (S 0 ). Perbedaan kandungan BK pada silase kontrol (S 0) dengan silase rumput gajah yang ditambahkan residu daun teh hitam (S 1 , S 2 , dan S3 ) dapat disebabkan kandungan BK residu daun teh hitam lebih tinggi dibandingkan dengan rumput gajah (55,48 vs. 19,94%). Disamping itu diduga peranan tanin yang menghambat pertumbuhan mikroorganisme seperti Clostridia sehingga menghambat degradasi nutrien selama ensilasi. Kandungan BK silase rumput gajah pada penelitian ini bervariasi antara 14,31 - 26,05%.

Nilai tersebut lebih tinggi dibanding hasil penelitian Kondo et al. (2004b) yang melaporkan bahwa kandungan BK silase rumput Sudan yang diberi penambahan residu daun teh hitam 50 g, 100 g dan 200 g/kg berat segar rumput dan tanpa bahan aditif bervariasi antara 20,70 - 22,20%. Pada penelitian lain, Kondo et al. (2004b) melaporkan bahwa kandungan BK silase rumput sudan yang ditambahkan residu teh hijau dengan konsentrasi 50 g dan 200 g/kg bahan segar dan diensilasi selama 30 hari bervariasi antara 27,70 29,80%. Kandungan BO silase rumput gajah meningkat secara linier (P<0,05) sejalan dengan peningkatan residu daun teh hitam dalam silase. Peningkatan ini disebabkan kandungan BO residu daun teh hitam (95,72%) lebih tinggi dibandingkan dengan BO rumput gajah (88,83%). Kandungan PK dari silase rumput gajah meningkat secara linier (P<0,01) dari perlakuan S0 sampai dengan S2 dan menurun mengikuti pola kuadratik (P<0,01) pada perlakuan S 3 . Kandungan PK pada perlakuan S 1 dan S2 mengalami peningkatan dibandingkan dengan perlakuan S 0 (kontrol) disebabkan kuantitas residu daun teh hitam pada silase tersebut meningkat. Residu daun teh hitam yang digunakan pada penelitian ini mengandung PK yang lebih tinggi dibandingkan rumput gajah (18,12% vs. 12,23%). Disamping itu, kandungan PK yang tinggi pada perlakuan S1 dan S2 dibandingkan S0 diduga berkaitan dengan peningkatan kandungan tanin sehingga menurunkan nilai degradasi PK pada perlakuan tersebut (Tabel 3). Menurut Bureenok et al. (2006); Tabacco et al. (2006), tanin dan polifenol dapat mengikat protein sehingga dapat memproteksi protein terhadap degradasi oleh mikroba selama ensilase. Perlakuan S 3 mengandung PK yang lebih rendah dibandingkan perlakuan S2 , dan hasil ini tidak sesuai dengan hipotesis pada awal penelitian. Hal ini diduga lebih disebabkan oleh faktor teknis seperti sampel yang digunakan dalam analisis tidak tercampur secara homogen. Data tersebut didukung pula oleh konsentrasi tanin pada perlakuan S 3 yang lebih rendah dibandingkan perlakuan S 2 . Kandungan PK pada silase yang ditambahkan 200 g residu daun teh hitam/kg rumput gajah (S2 ) mengalami peningkatan sebesar 49,70% dibandingkan perlakuan S 0 . Sementara pada penelitian Kondo et al. (2004b) dilaporkan bahwa dengan penambahan 200 g residu daun teh hitam/kg bahan silase, kandungan PK silase rumput sudah meningkat sebesar 104% dibandingkan silase kontrol. Dilaporkan pula bahwa

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kandungan PK residu daun teh hitam 27,60%, sehingga residu daun teh hitam berpotensi digunakan sebagai suplemen protein pada pengawetan hijauan pakan ternak dalam bentuk silase. Degradasi Bahan Kering, Bahan Organik dan Protein Kasar Selama Ensilase. Rata-rata degradasi BK, BO dan PK silase rumput gajah yang diensilase dengan residu daun teh hitam tertera pada Tabel 3. Nilai degradasi BK menurun secara linier (P<0,01) sejalan dengan penambahan residu daun teh hitam pada silase rumput gajah. Degradasi BK pada perlakuan S 1 , S2 dan S 3 menurun sebesar 24,49, 25,59 dan 51,05% dibanding perlakuan S0 (kontrol). Penurunan degradasi BK sejalan dengan penambahan residu daun teh hitam diduga karena kandungan BK bahan silase pada perlakuan tersebut semakin tinggi yaitu 19,94, 23,23, 28,50 dan 30,12% berturut-turut pada bahan silase S 0 , S1 , S 2 dan S 3 . Kondisi tersebut selanjutnya dapat menekan pertumbuhan Clostridia sakkarolitik yang terutama melakukan fermentasi terhadap karbohidrat dan asam organik. Menurut McDonald (1981), kandungan BK dari hijauan yang dibuat silase merupakan faktor yang paling mempengaruhi kuantitas BK yang hilang selama ensilasi. Pada penelitian menggunakan bahan silase alfalfa, Tabacco et al. (2006) melaporkan bahwa penambahan tanin chestnut

dengan konsentrasi 4% menurunkan nilai degradasi BK silase alfalfa dibandingkan dengan perlakuan kontrol (2,43 vs. 3,35%). Penurunan nilai degradasi BK tersebut disebabkan pola fermentasi yang baik dihasilkan dari penambahan tanin chestnut. Nilai degradasi BK silase rumput gajah yang diperoleh pada penelitian ini adalah berkisar antara 15,03 30,71%, dan lebih tinggi dibandingkan dengan nilai degradasi BK pada silase yang ideal yaitu 2 - 6% (Mc Donald, 1981) serta penelitian Kondo et al. (2006) yang melaporkan degradasi BK silase campuran limbah tahu, jerami padi dan dedak padi sebesar 10,50% sedangkan yang ditambahkan residu daun teh hijau sebesar 7,40%. Pada penelitian tersebut dilaporkan pula bahwa nilai degradasi BK silase yang disimpan pada suhu 30 0C lebih besar dibandingkan dengan silase yang disimpan pada suhu 15 0 C. Nilai degradasi BK silase rumput gajah dan rumput Itali yang diensilase selama 40 hari masingmasing 3,80% dan 7,30% (Yahaya et al., 2004). Nilai degradasi BK yang tinggi pada penelitian ini dibandingkan beberapa penelitian di daerah temperate dapat pula disebabkan oleh kandungan karbohidrat mudah larut dalam air pada rumput tropik lebih rendah sehingga konsentrasi asam laktat yang dihasilkan rendah dan tidak dapat menurunkan nilai pH dibawah nilai 4,00 (Smith, 1962; Yahaya et al., 2004).

Tabel 2. Kandungan BK, BO dan PK silase rumput gajah yang diensilase dengan residu daun teh hitam Perlakuan S.E. Signifikansi S0 S1 S2 S3 Linier Kuadratik BK (%) 14,31 18,12 22,33 26,05 0,64 ** TS BO (%) 82,37 87,36 88,23 89,60 0,92 ** TS PK (%) 9,56 12,36 14,31 12,93 0,41 ** ** Tanin (%) 0,03 0,04 0,04 0,03 S.E.: Standar error; TS : Tidak Signifikan (P>0,05); * : (P<0,05); ** : (P<0,01). BK: bahan kering, BO : bahan organik, PK: protein kasar. S 0 : silase rumput gajah saja (kontrol). S 1, S 2, S 3 , berurutan, silase rumput gajah + residu daun teh hitam 100, 200, 300 g/kg bahan segar.

Tabel 3. Nilai degradasi BK, BO dan PK silase rumput gajah yang diensilase dengan residu daun teh hitam S0 Degradasi BK (%) BO (%) PK (%) 30,71 35,70 46,07 S1 23,19 25,20 32,20 Perlakuan S2 22,.85 25,32 13,04 S.E. S3 15,03 20,45 8,88 2,86 3,51 3,44 Linier ** * ** Signifikansi Kuadratik TS TS TS

S.E. : Standar error; TS : Tidak Signifikan (P>0,05); * : (P<0,05); ** : (P<0,01). BK: bahan kering, BO : bahan organik, PK: protein kasar. S 0 : silase rumput gajah saja (kontrol). S 1 , S 2 , S 3, berurutan, silase rumput gajah + residu daun teh hitam 100, 200, 300 g/kg bahan segar.

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Penambahan residu daun teh hitam menurunkan nilai degradasi BO secara linier (P<0,05). Sementara itu, nilai degradasi PK menurun secara linier (P<0,01) hingga perlakuan S 2 (100 g residu daun teh hitam/kg bahan silase) (Tabel 3). Penurunan nilai degradasi PK sejalan dengan meningkatnya penambahan residu daun teh hitam, dapat disebabkan kandungan PK dan tanin yang meningkat hingga perlakuan S 2 , sebagaimana tertera pada Tabel 1. Menurut Salawu et al. (1999) penambahan tanin dalam pembuatan silase dapat menurunkan kandungan N yang mudah larut sehingga berpotensi sebagai agen proteksi protein selama fermentasi rumput. Pada percobaan skala laboratorium, Tabacco et al. (2006) melaporkan bahwa penambahan tanin chestnut 4 dan 6% menurunkan konsentrasi amonia dan NPN yang merupakan indikator proteolisis selama ensilase. Beberapa faktor penting yang mempengaruhi tingkat degradasi protein oleh enzim tanaman adalah kandungan BK, ada tidaknya oksigen, pH dan temperatur (Henderson, 1993). Tabel 1 memperlihatkan kandungan BK silase meningkat dari perlakuan S 0 sampai dengan perlakuan S 3, selanjutnya pola tersebut diikuti dengan penurunan nilai degradasi PK dari perlakuan S 0 sampai dengan perlakuan S 3 . Nilai degradasi PK pada perlakuan S 1 , S 2 dan S 3 yang lebih rendah dibandingkan perlakuan S 0 , diikuti pula dengan nilai degradasi BO yang rendah pada perlakuan tersebut. Hal ini disebabkan protein kasar merupakan salah satu komponen penyusun BO.

Daftar Pustaka
Bureenok, S., T. Namihira., S. Mizumachi., Y. Kawamoto and T. Nakada, 2006. The effect of epiphytic lactic acid bacteria with or without different byproduct from defatted rice bran and green tea waste onnapiegrrass (Pennisetum purpureum Shumach) silage Fermentation. Journal of the Science of Food and Agriculture 86: 10731077. Chamberlain, A.T. and J.M. Wilkinson, 1998. Feeding the Dairy Cows. Chalcombe Publications. Chamberlain, D.G., 1987. The silage fermentation in relation to the utilization of the nutrients in the rumen. Process Biochemistry 22: 6063. Crowder, L.V. and H.R. Chheda, 1982. Tropical Grassland Husbandry. Longman Group Limited, New York,. Pp. 562. Harris, L.E., 1970. Nutrition Research Techniques for Domestic and Wild Animals. Utah State University. Logan, Utah. Hartadi, H., S. Reksohadiprodjo dan A.D. Tillman, 1993. Tabel Komposisi Pakan untuk Indonesia. Gadjah Mada University Press, Yogyakarta. Henderson, N., 1993. Silage additives. Animal Feed Science and Technology 45 : 35 56. Kondo, M., K. Kita and H. Yokota, 2004a. Feeding value to goats of whole-crop oat ensiled with green tea waste. Animal Feed Science and Technology 113: 71-81. Kondo, M., K. Kita and H. Yokota, 2004b. Effects of tea leaf waste of green tea, oolong tea and black tea addition on sudan grass silage quality an in vitro gas production. Journal of the Science of Food and Agriculture 84: 721-727. Kondo, M., K. Kita and H. Yokota, 2006. Evaluation of fermentation characteristics and nutritive vaue of green tea waste ensiled with byproducts mixture for ruminants. Asian-Australian Journal of Animal Sciences 19: 533-540. Kondo, M., N. Naoki, K. Kazumi and H. Yokota, 2004c. Enhanced lactic acid fermentation of silage by the addition of green tea waste. Journal of the Science of Food and Agriculture 84: 728-734. McDonald, P., 1981. The Biochemistry of Toronto. Silage.

Kesimpulan
Kandungan BK dan BO silase rumput gajah meningkat secara linier, sedangkan kandungan PK meningkat secara kuadratik sejalan dengan peningkatan taraf residu daun teh hitam. Nilai degradasi BK, BO dan PK selama ensilase 30 hari menurun secara linier sejalan dengan penambahan residu daun teh hitam. Nilai degradasi BK bervariasi antara 15,03 - 30,71% dan lebih tinggi dibandingkan nilai degradasi BK silase yang ideal diduga karena kandungan BK dan karbohidrat mudah larut dalam air dari bahan silase yang digunakan rendah. Residu daun teh hitam berpotensi digunakan sebagai aditif silase untuk meningkatkan kualitas fermentasi serta suplemen protein pada pakan ruminansia.

Ohshima, M. and P. McDonald, 1978. A review of the changes in nitrogenous compounds of herbage

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during ensilage. Journal of the Science of Food and Agriculture 29: 497-505. Purnomo, H. 2004. Teh bisa cegah penyakit degeneratif. http//www. Media Indonesia Posted by roda bisnis at 8:49am o Commenents.htm. [13 Desember 2006]. Salawu, M.B., T. Acamovic and C.S. Stewart, 1999. The use of tannins as silage additives: effects on silage composition and mobile bag disappearance of dry matter and protein. Animal Feed Science and Technology 82: 243 254. Santoso, B., A. Kilmaskossu and P. Sambodo, 2006. Effects of saponin from Biophytum petersianum Klotzsch on ruminal fermentation, microbial protein synthesis and nitrogen utilization in goats. Animal Feed Science and Technology 137(1,2): 5868 SAS., 1996. SAS/STAT Software, Release 6,12. SAS Institute Inc, NC. Smith, L. H., 1962. Theoretical carbohydrate requirement for alfalfa silage production. Agronomy Journal 54 (4): 591-593. Sudirman, R. Utomo, Z. Bachrudin, B.P. Widyobroto and Suhubdy, 2006. An evaluation of in vitro method

using buffalo faeces as a source of inoculum for the measurement of tropical feed digestibility. In: H. Hartadi, K.A. Santosa, A. Wibowo, E. Suryanto, T. W. Murti, B. Rustamadji, J. H. P. Sidadolog, C. T. Noviandi (Eds.). Proceedings of the 4th International Seminar on Tropical Animal Production. Yogyakarta, November 8-9, 2006. Pp. 135-139. Tabacco, E., G. Borreani and G. M. Crovetto, 2006. Effect of chestnut tannin on fermentation quality, proteolysis and protein rumen degradability of alfalfa Silage. Journal of Dairy Science 89: 47364746. Yahaya, M.S., M. Goto, W. Yimiti, B. Smerjai and Y. Kuwamoto, 2004. Evaluation of fermentation quality of a tropical and temperate forage crops ensiled with additives of fermented juice of Asianepiphytic lactic acid bacteria (FJLB). Australian Journal of Animal Science 17: 942-946. Yahaya, M. S., M. Kawai, J. Takahashi and S. Matsuoka, 2002. The effects of different of moisture content at ensiling on silo degradation and digestibility of structural carbohydrates of orchard grass. Animal Feed Science and Technology 101: 127-133.

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Vol. 9 No.3

The Effect of Increasing Levels of Whole Cottonseed Supplementation on Microbial Protein Production of Steers Fed Low Quality Forage
(Pengaruh Suplementasi Biji Kapas terhadap Produksi Protein Mikroba Rumen pada Sapi Jantan yang Diberi Pakan Rumput Berkualitas Rendah)
Marsetyo*
Faculty of Agriculture, Tadulako University Palu, Central Sulawesi

ABSTRAK: Pengaruh taraf suplementasi biji kapas utuh terhadap produksi protein mikroba dan efisiensi produksi protein mikroba dalam rumen pada sapi jantan bakalan yang mendapatkan rumput berkualitas rendah telah dikaji dalam penelitian ini. Sebanyak lima ekor sapi jantan bakalan Charbray persilangan dialokasikan secara random terhadap lima taraf biji kapas utuh. Rancangan percobaan yang digunakan adalah bujur sangkar latin (5x5), yang masing-masing dengan lima periode sebagai ulangan. Dalam satu periode terdiri atas 21 hari dimana 14 hari untuk masa adaptasi dan 7 hari untuk pengukuran. Sapi percobaan mendapatkan pakan dasar berupa hay Chloris gayana secara ad libitum dan mendapatkan suplementasi biji kapas utuh dengan taraf 0; 0,15; 3,00; 4,50; dan 6,00% dari bobot badan per hari (BB/hari). Hasil penelitian menunjukkan bahwa taraf suplementasi biji kapas utuh pada sapi yang mengkonsumsi rumput berkualitas rendah berkorelasi secara kuadratik terhadap produksi protein mikroba (P<0,01) dan efisiensi produksi protein mikroba di dalam rumen (P<0,05). Pada kedua parameter tersebut, produksi protein mikroba dan efisiensi produksi protein mikroba tertinggi dihasilkan pada suplementasi biji kapas utuh pada taraf 0,30% BB/hari. Konsentrasi nitrogen amonia dari cairan rumen juga berkorelasi secara kuadratik terhadap peningkatan taraf suplementasi biji kapas utuh (P<0,05). pH cairan rumen tidak mengalami perubahan seiring dengan peningkatan suplementasi biji kapas utuh (P>0,05). Dapat disimpulkan bahwa produksi protein mikroba dan efisiensi produksi protein mikroba tertinggi diperoleh pada penambahan biji kapas utuh dengan taraf sedang. Kata Kunci: Biji kapas utuh, sapi bakalan, protein mikroba, nitrogen ammonia

Introduction
Interest in the use of whole cottonseed (WCS) as a component of rations for ruminants has increased. WCS contains high levels of fibre, lipid and moderate levels of protein which makes it a valuable supplement for beef and dairy cattle (McLennan et al., 1998; Meyer et al., 2001; Johnson et al., 2002; Noftsger et al., 2000; Chen et al., 2007a). However, the degree of ruminal crude protein (CP) degradation of WCS appears to be controversial, which may affect the animal's productivity. Some studies have indicated a high ruminal CP degradability which ranges between 74 and 80% (Whitney et al., 2000; Enjalbert et al., 2003; Chen et al., 2007b). Others, however, have found a low ruminal CP degradability (Baldwin and Allison, 1983; Palmquist, 1995; Whitney et al., 2000; Harvatine et al., 2002; Reveneau et al., 2005). Mullik (1999) recorded a high CP degradability in

the rumen (68%) in a nylon bag trial, but there was no significant difference in the rumen NH 3-N concentration between control and WCS supplemented steers. Similarly, inclusion of WCS up to 20% of the total diet failed to increase rumen NH 3-N level in the studies of Whitney et al. (2000). The high lipid content of WCS is presumably one factor which may inhibits access to the site and degradation of protein (Bottger et al., 2002; Palmquist, and Jenkins, 2003; Moreira et al., 2004). As a consequence, WCS supplementation to forage tended to depress microbial protein production in the rumen (McLennan et al., 1998; Oldick and Firkins, 2000). The objectives of this experiment was to examine the effect of increasing levels of WCS intake on microbial protein production and efficiency of microbial protein production in the rumen of cattle fed low quality forage.

Research Methods
Treatments and Feeding
Five Charbray crossbred steers approximately 205 10.6 (SE) kg in initial weight and 12 months

Corresponding author e-mail: marsetyo@yahoo.co.uk

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of age were used. Steers were initially adapted to diets and surroundings for 21 d prior to the start of the preliminary period. During this period, the intake of WCS was gradually increased to avoid any digestive problems. At the beginning of this period, the steers were injected with Ivomec (10 g/L Ivermectin, Merck and Co. Inc. White House Station, New Jersey, USA) for the control of internal and external parasites. A 5 x 5 Latin square, with five runs, i.e., five steers/treatment overall, was employed in the study. The treatments were five levels of feeding equivalent to 0.0, 0.15, 0.30, 0.45 and 0.60% W/d for WCS (on dry matter basis). The basal diet was Rhodes grass hay which was given individually to each steer ad libitum. Each run lasted for 21 days. The steers were fed their diet in individual pens over a 14 d preliminary period and then transferred to individual metabolism crates for 7 d collection period. The steers were weighed at the beginning of the preliminary and collection period to adjust the WCS allocation. The hay offered was chopped into lengths of 5 to 13 cm. Steers were fed a recorded weight of hay allowing a 10-15% excess of hay based on the hay intake of the previous 24 h. The WCS allocation was offered separately to the basal diet at 0700h (1 h before hay was offered). The whole seed was partially delinted by the Company. Water and a mineral block (no N) was freely available at all times. The composition of the mineral block was as follows: salt (53.6%), molasses (5%), calcium (13.5%), phosphorus (0.6%), sulphur (0.8%), copper (0.1%), cobalt (0.006%), magnesium (0.02%), ferrous iron (Fe++, 0.135%), iron (Fe+++, 0.065%) and fluorine (F, 0.09%).

high performance liquid chromatography procedure. The MCP production was estimated by using the formula of Bowen (2003). The efficiency of MCP production (eMCP; g CP/kg DOM) was calculated based on the value of MCP production (g CP) divided by total digestible organic matter intake (TDOMI; kg ). Samples of rumen fluid (at least 100 mL) were taken 3 h after feeding on day 7 of the collection period, by inserting a plastic tube down the oesophagus and into the rumen and withdrawing a sample using a vacuum pump. Ruminal pH was measured on fresh fluid immediately after sampling and a sub-sample (20 mL) of rumen fluid for chemical analysis was placed immediately into two and a sub-sample (20 mL) of rumen fluid for chemical analysis was placed immediately into two tubes (10 mL capacity), each caontaining 0.2 mL of concentrated H 2SO4, and stored at 20 0 C prior to determination of NH 3 -N concentration.

Chemical analysis
Samples of feed, refusals and feces were analysed for dry matter (DM) and organic matter (OM) according to AOAC (2000) procedures on ground samples (1 mm screen). The nitrogen was determined using an automatic total nitrogen analyzer (LECO FP-428). Neutral detergent fibre was determined by procedures of Goering and Van Soest (1970) using a fibre extraction unit (ANKOM 220). The ether extract content of the hay and supplements was analysed using a solvent extraction unit (Soxtec HT6, Tecator, Sweden). The concentration of NH 3 N in the rumen fluid was determined by a distillation method using a Buchi 321 distillation unit (Buchi Scientific Apparatus Flawil, Switzerland). The urine samples were analysed for purine derivatives using high performance of liquid chromatography procedures.

Measurements
Microbial crude protein (MCP) production in the rumen was estimated from the excretion of purine derivatives in urine which was collected dailyduring the collection period into trays below the metabolism crate. The pH of the urine was kept just below 3.0 by adding 10% H 2 SO 4 into individual trays at the start of each daily collection. For each steer, urine collection over a 24 h period was mixed and a 5% aliquot was taken, bulked over the collection period and stored in a refrigerator. At the end of each collection period, a 5 mL sub-sample was taken from the bulked samples from each steer, diluted to 50 mL with ammonium phospate stock buffer and frozen before purine derivatives (PD) analysis. The urine was analysed for PD by using a

Parameters Measured
The parameters measured in this experiment were microbial crude protein production in the rumen, efficiency of microbial crude protein production in the rumen, rumen NH 3 -N concentration at 3 h after supplement feeding and rumen pH at 3 h after supplement feeding.

Statistical Analysis
The data obtained were analyzed by using fitted general linear models with level of WCS intake as

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Table 1. Chemical composition of feed ingredients used in the experiment (% dry matter) Organic matter (%) Crude protein (%) Neutral detergent fibre (%) Ether Extract (%) Rhodes grass hay 91.00 5.80 74.70 1.40 Whole cottonseed 95.20 22.40 51.10 20.90

an effect from Genstat 6 th edition program (Lawes Agricultural Trust, 2002). Polynomials were used to describe the response of parameters measured to the level of supplement intake.

Results and Discussion


Microbial Protein Production
The MCP production and eMCP in the rumen observed in this experiment are shown in Table 2. Supplementation of WCS resulted in a significant quadratic increase (P<0.05) in both MCP production and eMCP which peaked at 0.30 of WCS intake. The mean values of MCP production and eMCP of control steers over the three runs were 179 g MCP/d and 95 g MCP/kg DOM respectively. Based on the total DOM intake, the theoretical MCP supply of the control steers in the present study should be approximately around 232 g MCP/d (SCA, 1990). It is apparent that MCP supply of control steers reached only 78% of the theoretical values recommended by SCA (1990). The low level of rumen degradable protein (RDP) estimated as 90 g RDP/kg DOM based on the forage CP degradability (75%) of McLennan et al. (1997) was reflected in the low rumen NH 3-N concentration. The rumen NH 3-N concentrations of control steers at 3 h after feeding were very low (mean value of 32 mg/L) and below the recommended level of 50 mg/L (Satter and Slyter, 1974). Both MCP production and eMCP were affected quadratically (P<0.01) by increasing level of WCS inclusion in the diet. The data show that the maximum amount of MCP production (343 g MCP/d) and eMCP (111 g MCP/kg DOM) were achieved when the steers received approximately 0.40% W/d of WCS which was a little above the value to maximise intake and digestibility. It is most likely that the increase in MCP production and eMCP may be attributed to an increase in RDP supply and an increase in total DOM intake. However, as discussed earlier, RDP supply was still most likely limiting and could not be increased without depressing forage intake. However even when the CP/DOM was increased, despite lower

forage intake, the eMCP was still less than 130 g MCP/kg DOM suggesting that the high lipid level also depressed eMCP as well as intake. Both MCP production and eMCP started to decline when the level of WCS feeding exceeded approximately 0.40% W/d. The depression in eMCP due to high level of WCS agrees with other earlier studies (Reveneau et al., 2005; Sullivan et al., 2005) which showed that inclusion of WCS up to 25.30% in the diet decreased eMCP by up to 21%. There may have been dilution rate changes associated with this as Bowen (2003) showed these changed directly with changes in intake but there may have been effects of high lipid as well (Allen, 2000; Onetti et al., 2001; Bauman and Griinari, 2003; Enjalbert et al., 2003; Lima et al., 2003). Although the eMCP value of WCS supplementation increased quadratically (P<0.05), the maximum value reached at 0.40% W/d of WCS was 111 g MCP/kg DOM was below the lower range value recommended by the current feeding standards (SCA, 1990; NRC, 2001). The SCA (1990) and NRC (2001) for example, suggest that the values should range between 130 and 170 g MCP/ kg DOM. With the current basal diet (5.8% CP), assuming that the degradability of CP in the rumen of Rhodes grass hay is 75% (McLennan et al., 1997) and WCS is 68% (Mullik, 1999), the availability of RDP with 0, 0.15, 0.30, 0.45 and 0.60% W/d of WCS intake was 90, 101, 106, 132 and 163 g RDP/kg DOM, respectively. It was clear that at all level of WCS intakes except at the second highest level, the RDP availability was below 130 g RDP/kg DOM recommended by SCA (1990). Based on RDP availability, at 0.60% W/d of WCS intake, the eMCP should theoretically show the highest value and above the recommended level by SCA (1990) but it was only 81% of the lower value recommended by SCA (1990). There are probably some reasons to explain the low eMCP from WCS. Firstly, there is the high lipid content of WCS used in the current experiment. Earlier studies (Baldwin and Allison, 1983; Mabjeesh et al., 2000; Oldick and Firkins, 2000; Mosley et al., 2002; Santos et al., 2002) have

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Table 2. Effect of increasing whole cottonseed intake on microbial protein production, efficiency of microbial protein production in the rumen and other rumen parameters on cattle given low quality forage Parameters MCP production (g MCP/d) eMCP (g MCP/kg DOM) Rumen NH 3-N 3 h (mg/L) Rumen pH-3h 0 179 14.20 95 4.81 35 3.16 7.01 0.06 WCS DM intake (%W/d) 0.15 0.30 0.45 237 14.20 343 14.20 296 14.20 99 4.81 111 4.81 105 4.81 65 3.16 75 3.16 86 3.16 7.03 0.06 7.17 0.06 7.00 0.06 Probability Linear Quadrat <0.01 <0.01 <0.01 0.02 <0.01 0.04 0.57 0.74

0.60 262 14.20 105 4.81 93 3.16 7.13 0.06

MCP: microbial crude protein, eMCP: efficiency of MCP, WCS: whole cotton seed, DM: dry matter

observed that the fatty acids of WCS may protect the seed coat from microbial biohydrogenation. Mullik (1999) indicated that supplementation of steers consuming Pangola grass hay with 0.50% W/d WCS only obtained an eMCP value of 65 g MCP/kg DOM which is well below the current result. The second reason is the type of basal diet used in the current experiment. The rumen dilution rate of these tropical grasses is low as is the supply of soluble carbohydrates (Slater et al., 2000), and this will set eMCP at a lower value but the literature indicates this should approximate 130 g MCP/kg DOM (SCA, 1990; NRC, 2001). A third possible reason is the low intake observed in the current experiment. Total DM intake ranged from 1.42 to 2.02% W/d (Marsetyo et al., 2002).

2). The overall mean values for pH in rumen fluid (taken 3 h after feeding) were 7.07 0.03, and appeared within the normal range for rumen fermentation (6.5 to 7.0) (Baldwin and Allison, 1983; Gonthier et al., 2004; Solomon et al., 2005; Sullivan et al., 2005; Suarez, 2006).

Conclusion
It was concluded that steers would readily consume up to 0.60% W/d of WCS but that the optimum level of WCS supplementation appeared to be 0.30-0.40% W/d. With this medium level of WCS supplementation, MCP production and eMCP reached maximum levels. It was also suggested that providing more RDP from another source (e.g. urea) might further enhance the intake and MCP response of cattle supplemented with WCS.

NH3-N and pH in the Rumen Fluid


The concentrations of NH 3 -N and pH in the rumen fluid of steers are shown in Table 2. The concentration of NH 3 -N in rumen fluid was increased (P<0.05) by WCS supplementation. The mean rumen NH 3 -N concentrations of control steers over the three runs were 35 mg/L for samples taken 3 h after feeding. These levels were clearly insufficient to meet microbial requirements (Satter and Slyter, 1974). Apart from the control group, rumen NH 3 -N concentration of samples taken 3 h feeding for all supplemented steers were above 50 mg/L, which is the minimum required for maximal microbial synthesis (Satter and Slyter, 1974). Ammonia is the main nitrogen sources for microbial protein synthesis in the rumen. Bryant and Robinson (1962) noted that more 80% of a crosssection of 89 ruminal species examined grew with ammonia as the major source of nitrogen. Therefore, ensuring the presence of adequate ammonia in the rumen to supply nitrogen for microbial growth is generally considered to be the first priority in optimizing fermentative digestion of forage. In contrast, WCS supplementation had no significant effect on the pH of rumen fluid (Table

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Reveneau, C. C.V.D.M. Ribeiro, M.L. Eastridge, N.R.StPierre, and J.L. Firkins, 2005. Processing whole cottonseed moderates fatty acid metabolism and improves performance by dairy cows. Journal Dairy Science 88: 4342-4355. Santos, J.E.P., M. Villasenor, E.J. DePeters, P.H. Robinson, and B.C. Baldwin Jr, 2002. Type of cottonseed and level of gossypol in diets of lactating dairy cows: Effects on lactation performance and plasma gossypol. Journal of Dairy Science 85: 1491-1501. Satter, L.D. and L.L. Slyter, 1974. Effect of ammonia concentration on rumen microbial protein production in vitro. British Journal of Nutrition 32:199-208. [SCA] Standing Committee on Agriculture, 1990. Feeding Standards for Australian Livestock: ruminants. CSIRO, Melbourne, Australia. Slater, A.L., M.L. Eastridge, J.L. Firkins, and L.J. Bidinger, 2000. Effect of starch source and level of forage neutral detergent fibre on performance by dairy cows. Journal of Dairy Science 83: 313-321.

Solomon, R., G. Adin, S.J. Mabjeesh, M. Nikbachat, E. Yosef, D. Ben-Ghedalia, and J. Miron, 2005. Digestibility in lactating cows of diets containing whole Pima treated with sodium hydroxide versus Akala or Pima cottonseed. Journal of Dairy Science 88: 1745-1751. Suarez, B.J, C.G. Van Reenen, G. Beldman, J. Van Delen, J. Dijkstra, W.J.J. Gerrits, 2006. Effects of supplementing concentrates differing in carbohydrate composition in veal calf diets: I. Animal performance and rumen fermentation characteristics. Journal of Dairy Science 89: 43654375. Sullivan, H.M., J.K. Bernard, and H.E. Amos, 2005. Ruminal fermentation and amino acid flow in Holstein steers fed whole cottonseed with elevated concentration of free fatty acids in the oil. Journal of Dairy Science 88: 690-697. Whitney, M.B., B.W. Hess, L.A. Burgwald-Balstad, J.L. Sayer, C.M. Tsopito, C.T. Talbott, and D.M. Hallford, 2000. Effects of supplemental soybean oil level on in vitro digestion and performance of pre-pubertal beef heifers. Journal of Animal Science 78: 504-514.

ANIMAL PRODUCTION, September 2007, hlm. 172 177 ISSN 1411 2027 Terakreditasi No.56/DIKTI/Kep/2005

Vol. 9 No.3

Pengaruh Ekstrak Benalu Teh (Scurrula oortiana) sebagai ImunoModulator pada Infeksi Mareks Disease Virus Onkogenik
(The effect of Tea Misletoe (Scurrula oortiana) Stem Extract as Immuno-Modulator on Oncogenic Mareks Disease Virus Infection)
Mohamad Samsi1* , Marthen Benedictus Melkianus Malole2, Wasmen Manalu2 dan Ekowati Handharjani2
1 2

Fakultas Peternakan Universitas Jenderal Soedirman, Purwokerto Fakultas Kedokteran Hewan Institut Pertanian Bogor, Bogor

ABSTRACT: Mareks disease virus (MDV) is one of oncogenic herpesvirus. It causes immunosupresion and cancer in chicken. Several plants produce bioactive compounds which are very useful for treatment of many disease, especially hiperproliveration and virus infection. This study was aimed to find out mechanism of immuno-modulatory capacity in layer commercial chicken administered orally with extract of tea parasite (Scurrula oortiana) in dose of 10 mg/kg BW through drinking water, then the chicken were infected by intraperitoneal oncogenic MDV in dose of 1,0 x103 TCID 50. The study used 60 layer commercial day old chicks (DOC) divided into four group treatments. The treatments were group A (administered S. oortiana extract and without MDV infection), B (neither S. oortiana nor MDV infection), C (administered S. oortiana extract and with MDV infection), and D (without administered S. oortiana extract, but with MDV infection). Results showed that MDV oncogenic caused immunosupresion at a day post infection (p.i) and recovery to be normal based on relative weight of bursa Fabricius and thymus at 40 days p.i. The extract of S. oortiana had a capability as an immunomodulator indicated by the increase of relative weight of bursa Fabricius and thymus at day 20 days p.i. Key Words: Mareks disease virus (MDV), Scurrula oortiana, immuno-modulator

Pendahuluan
Mareks disease (MD) disebabkan oleh virus DNA termasuk pada group virusherpes- penyebab kanker pada ayam. Virus tumbuh dan berkembang pada epitelium folikel bulu kemudian menyebar ke udara selanjutnya menular melalui ketombe dan debu (Silva et al., 2004). Target pertama diantaranya adalah derivat bursa Fabricius (limfosit B), namun sejumlah derivat timus (limfosit T) juga mengalami infeksi. Selama 3 sampai 6 atau 7 hari pascainfeksi (p.i.) terjadi infeksi sitolisis, pembesaran limpa, disertai nekrosis dan atrofi bursa Fabricius dan timus (Calnek et al., 1998). Mareks disease virus (MDV) isolat Austalia MPV 57 menimbulkan imunosupresi pada ayam pedaging bersamaan dengan turunnya bobot relatif bursa Fabricius dan timus, dan peningkatan kepekaan pada infeksi Escherichia colli (Islam et al., 2002).

Penulis Korespondensi. e-mail: Zamsyvet@yahoo.com

Diet antioksidan eksogen mencegah kerusakan seluler (sitolisis) melalui reaksi yang dilakukan oleh radikal bebas. Ayam yang diberi pakan diet semisintetik rendah antioksidan menunjukkan penurunan stabilitas eritrosit terhadap H2 O 2 tetapi terjadi peningkatan pada aktivitas katalase pada hepar, karbonil pada protein otot tak larut (Young et al., 2002). Antioksidan yang berasal dari tanaman telah lama dikenal potensinya dan telah lama diketahui untuk menstabilkan senyawa radikal yang dapat diukur aktivitas antioksidan tersebut (Kim et al., 2002). Benalu teh secara tradisional digunakan untuk penyembuhan berbagai penyakit diare, kanker, dan amandel. Beberapa publikasi hasil penelitian telah melaporkan efek benalu teh diantaranya sebagai perbaikan sistem imun (Winarno et al., 2003), dan hambatan pertumbuhan sel tumor (Nugroho et al., 2000). Tanaman benalu teh (di benua Eropa disebut Viscum album L.) yang dalam percobaan bersifat imunostimulator melalui pengaktifan sel granulosit dan makrofag yang memberi sifat anti tumor (Achi 2005). Daun dan batang benalu teh mengandung senyawa alkaloid, flavonoid, trepenoid, glikosida, triterpen, saponin, dan tanin (Nugroho et al., 2000 dan Tambunan et al., 2003).

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Senyawa polifenolat alam, termasuk flavonoid yang disintesis oleh tanaman, mampu memperbaiki kesehatan. Kuersetin dan kuersetin glikosida yang tersebar pada flavonoid banyak ditemukan pada buah dan sayur. Senyawa ini secara luas berperan pada perbaikan kesehatan sehingga menjadi penting dan menarik (Boyer et al., 2005, Lila et al., 2005). Ikatan dengan protein menghasilkan pelapisan substansi yang merupakan kapasitas antioksidan flavonoid. Pada kejadian ini penambahan aktivitas intrinsik dari senyawa, metabolisme, ikatan terhadap protein juga menentukan untuk mempengaruhi efek pemberian flavonoid secara invivo (Arts, 2002). Penelitian ini bertujuan untuk mengembangkan mekanisme imunomodulator dari benalu teh, menggunakan parameter bobot relatif organ limfoid yaitu bursa Fabricius, timus, dan limpa dari ayam ras petelur yang diinfeksi MDV onkogenik. Hasil penelitian ini diharapkan dapat memberikan informasi tentang potensi benalu teh strain Scurrula oortiana mencegah imunosupresi pada ayam yang diinfeksi MDV onkogenik.

Metode Penelitian
Penelitian dilaksanakan di kandang percobaan unggas Fakultas Kedokteran Hewan, Institut Pertanian Bogor (IPB). Pengamatan efek patologi anatomi dilaksanakan di Laboratorium Patologi Fakultas Kedokteran IPB. Ayam percobaan adalah ayam ras petelur strain Isa Brown yang diperoleh dari peternakan pembibitan Manggis Farm desa Tenjoayu Sukabumi, Jawa Barat. Benalu teh spesies Scurrula oortiana diperoleh dari Perkebunan Teh PTP Rancabuli, Cibuni, Bandung dan ekstraksi dilakukan di Laboratorium Bahan Makanan Ternak, Universitas Diponegoro, Telukawur Jepara Jawa Tengah, ekstraksi dengan metode reflux menggunakan air sebagai pelarut (Murtini, 2006). Virus Marek onkogenik serotipe 1 yang digunakan dalam penelitian ini diperoleh dari Balai Besar Pengawasan Mutu dan Sertifikasi Obat Hewan (BPMSOH) Direktorat Jenderal Peternakan, Departemen Pertanian, Gunung Sindur Bogor. Ayam percobaan secara acak ditempatkan dalam kandang, adaptasi ayam percobaan tidak dilakukan karena menggunakan ayam umur satu hari (day old chicken DOC). Kandang percobaan yang digunakan adalah sistem group cages berukuran 60 x 45 x 30 cm, masing-masing unit terdiri atas 3 ekor sehingga jumlah kandang seluruhnya 20 unit. Pada penelitian ini digunakan 60 ekor ayam dibagi ke

dalam empat kelompok perlakuan yaitu : perlakuan A. diberi ekstrak S. oortiana tanpa infeksi MDV, B tanpa pemberian ekstrak S. oortiana dan tanpa infeksi MDV, C diberi ekstrak S. oortiana dan diinfeksi MDV, dan D tanpa diberi ekstrak S.oortiana, diinfeksi MDV. Ekstrak benalu teh diberikan secara oral (dicekok) sejak ayam berumur 15 hari sampai akhir percobaan, dengan dosis 10 mg/kg bobot badan yang dilarutkan dalam air minum. Ayam diinfeksi dengan virus Marek pada umur 20 hari secara intraperitoneal (Cho et al., 1999) dengan dosis 1.000 TCID 50 (total count infectious dosis 50). Untuk menentukan bobot relatif bursa Fabricius, timus, dan limpa dilakukan penimbangan bobot badan ayam. Kemudian dilakukan bedah bangkai dan penimbangan bobot organ bursa Fabrisius, timus, dan limpa, kemudian hasil penimbangan bobot organ tersebut dibagi dengan hasil penimbangan bobot badan masing-masing ayam, sehingga didapatkan bobot relatif bursa Fabricius, timus, dan limpa. Penelitian dilakukan dengan menggunakan Rancangan Acak Lengkap (RAL). Data dianalisis dengan Analisis Variansi dan uji lanjut Kontras Ortogonal (Steel dan Torrie, 1991) menggunakan program SPSS versi 10 (SPSS, 1999).

Hasil dan Pembahasan


Pengaruh Ekstrak Benalu pada Bobot Relatif Organ Limfoid 20 Hari Pasca Infeksi
Kinerja sistem imun juga dapat diukur dari bobot relatif organ limfoid. Bursa Fabricius berperan pada pematangan limfosit B dan timus berperan pada pematangan limfosit T, yang merupakan organ limfoid primer. Infeksi MDV pada ayam diawali dengan periode infeksi sitolisis produktif, MDV menginfeksi limfosit B pada bursa Fabricius maupun limfosit T pada timus, terjadi replikasi DNA, sintesis protein, dan perbanyakan partikel virus. Pada puncak infeksi terjadi sitolisis dan kematian sel, atropi pada bursa fabricius dan timus sehingga terjadi imunosupresi, turunnya bobot relatif organ limfoid bursa Fabricius, dan timus yang dapat dijadikan sebagai indikator imunosupresi sebagai akibat dari infeksi MDV. Periode infeksi MDV meliputi 3 bentuk, yaitu infeksi akut (produktif) yang menimbulkan lisis sel, dilanjutkan infeksi laten yang bersifat nonproduktif, dan infeksi transforming. Pada infeksi produktif

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terjadi replikasi DNA virus, sintesis protein, dan menghasilkan partikel virus. Virus menginfeksi dan merusak limfosit B maupun limfosit T. Selama infeksi terjadi sitolisis pada puncak replikasi virus sehingga menyebabkan imunosupresi, dan meningkat kepekaan terhadap infeksi, bersamaan dengan turunnya bobot relatif bursa Fabricius dan timus (Calnek et al., 1998, Payne dan Venugopal, 2000, Islam et al., 2002). Replikasi virus herpes pada bursa Fabricius dan timus menimbulkan transien imunosupresi, perubahan sitolitik akut pada organ ini ditandai dengan atropi. Infeksi eksperimental terjadi lesi bursa Fabricius mengalami degenerasi folikuler, nekrosis limfoid sehingga mengalami atrofi, dan pembentukan kista. Timus mengalami atrofi, limfosit hilang baik pada kortek maupun medula. Benda inklusi intranuklear dapat muncul pada sel yang mengalami degenerasi (Fadly, 2000). Rataan bobot relatif organ bursa Fabricius 20 hari p.i pada berbagai kelompok perlakuan benalu teh dan infeksi MDV disajikan pada Tabel 1. Hasil analisis statistik menunjukkan adanya perbedaan yang signifikan (P<0,05) diantara fraksi kelompok perlakuan pada hari ke 20 p.i. Kelompok perlakuan dengan pemberian benalu teh tanpa infeksi MDV (A) memiliki nilai tertinggi sebesar 0,0037 berbeda dengan kelompok dengan pemberian benalu dan infeksi MDV (C) memiliki nilai 0,0022, dan juga berbeda dengan perlakuan yang tanpa diberi benalu teh dan diinfeksi MDV (D), yaitu 0,0021. Tingginya ratio bobot bursa Fabricius pada perlakuan A disebabkan oleh pengaruh imunomodulator dari ekstrak benalu teh spesies S. Oortiana 10 mg/kg bobot badan. Rendahnya bobot relatif bursa Fabricius pada kelompok perlakuan C dan D disebabkan oleh infeksi produktif yang menimbulkan sitolisis MDV pada 20 p.i. Pada Gambar 1 disajikan rataan bobot relatif bursa Fabricius pada 20 hari dan 40 hari pascainfeksi.

Terjadinya imunomodulasi pemberian ekstrak benalu teh spesies S. oortiana pada kelompok ayam tanpa infeksi MDV perlakuan A ditandai dengan perbaikan performan bursa Fabricius berdasarkan bobot relatif organ tersebut, dan kecenderungan terjadinya imunosupresi pada kelompok ayam yang diinfeksi MDV baik yang diberi ekstrak S. oortiana maupun tanpa diberi ekstrak S. oortiana. Kelompok perlakuan B, yaitu tanpa diberi benalu dan tanpa infeksi MDV adalah 0,031 tidak berbeda dengan semua kelompok perlakuan. Hal ini menunjukkan belum ada pengaruh imunosupresi pada ayam yang diinfeksi MDV pada 20 hari p.i, baik yang diberi benalu teh maupun tanpa diberi benalu teh. Hasil analisis statistik bobot relatif timus menunjukkan adanya perbedaan yang signifikan (P<0,05) di antara kelompok perlakuan pada hari ke 20 p.i. Kelompok perlakuan yang diberi benalu teh tanpa infeksi MDV (A) memiliki nilai tertinggi sebesar 0,0054 tidak berbeda dengan kelompok yang diberi perlakuan tanpa diberi benalu dan tanpa infeksi MDV (B) sebesar 0,0053. Kelompok A dan B berbeda dengan perlakuan D yaitu tanpa diberi benalu teh diinfeksi MDV (0,0019). Hal ini menunjukkan bahwa perlakuan infeksi MDV menimbulkan imunosupresi dilihat dari turunnya bobot relatif timus. Imunodifisiensi mungkin disebabkan oleh cacat pada pendewasaan limfosit atau aktifasinya atau gangguan pada mekanisme efektor imunitas alami maupun imunitas perolehan. Proses pendewasaan limfosit dari sel stem ke komponen sel fungsional limfosit dewasa termasuk proliferasi, ekspresi reseptor antigen, seleksi sel sehingga memiliki spesifitas, dan perubahan pada ekspresi sejumlah gen (Abbas et al., 2000).

Tabel 1 Rataan bobot relatif bursa fabricius, timus, dan limpa 20 hari p.i. Peubah Bursa Fabricius Timus Limpa A a 0,0037 0,0003 0,0054 a 0,0007 0,0039 a 0,0007 Perlakuan B ab 0,0031 0,0002 0,0053 a 0,0003 0,0034 a 0,0004 C 0,0022 0,0008
b

D 0,0021 0,0009
b

0,0033 ab 0,0025 0,0042 a 0,0011

0,0019 b 0,0003 0,0029 a 0,0010

a,b Superskrip dengan huruf yang berbeda pada baris yang sama menunjukkan ada perbedaan pada P<0,05 A = diberi ekstrak S. oortiana tanpa diinfeksi MDV B = tanpa diberi ekstrak S. oortiana tanpa diinfeksi MDV C = diberi ekstrak S. oortiana dan diinfeksi MDV D = tanpa diberi ekstrak S. oortiana diinfeksi MDV

Pengaruh Ekstrak Benalu Teh (Samsi et al.)

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A = diberi ekstrak S. oortiana tanpa diinfeksi MDV B = tanpa diberi ekstrak S. oortiana tanpa diinfeksi MDV C = diberi ekstrak S. oortiana dan diinfeksi MDV D = tanpa diberi ekstrak S. oortiana diinfeksi MDV

Gambar 1. Bobot relatif bursa fabricius pada 20 hari dan 40 hari pascainfeksi

Kriteria dari imunosupresif meliputi: (1) kejadian awal infeksi sitolisis, (2) atropi bursa fabricius dan timus yang diukur dari proporsi bobot organ limfoid terhadap bobot tubuh pada 8-14 pascainfeksi (p.i), dan (3) perubahan histopatologi yaitu nekrosis dan atropi organ limfoid. Disimpulkan bahwa tingkat imunosupresi adalah berhubungan dengan virulensi dan ukuran organ yang mengalami perubahan atrofi bursa Fabricius dan timus dapat digunakan sebagai pengukuran patotipe pada isolat baru MDV (Calneck et al., 1998). Kelompok dengan pemberian benalu teh spesies S. oortiana dan diinfeksi MDV (C) memiliki nilai 0,0033 tidak berbeda dengan semua kelompok perlakuan baik A, B, maupun D. Hal ini menunjukkan bahwa perlakuan pemberian ekstrak benalu teh mampu menghambat proses terjadinya sitolisis pada timus akibat infeksi sitolitik MDV. Perlakuan C tidak terpengaruh adanya imunosupresi yang disebabkan oleh MDV diimbangi oleh pengaruh imunomodulasi oleh ekstrak benalu teh spesies S. oortiana. Karena itu efek protektif dari antioksidan pada pencegahan kerusakan membran sel dan reseptor terhadap peroksidasi lipid dapat memberikan keuntungan pada perbaikan kinerja sistem imun. Pada Gambar 2 disajikan rataan bobot relatif timus pada 20 hari dan 40 hari pascainfeksi. Adanya imunomodulator berdasarkan bobot relatif bursa Fabricius pada pemberian ekstrak benalu teh tanpa infeksi MDV dan imunomodulator berdasarkan bobot relatif timus pada kombinasi

pemberian benalu teh dan disertai infeksi MDV Maka ekstrak benalu teh spesies S. oortiana mampu memperbaiki performan sistem imun organ limfoid primer baik pada bursa Fabricius maupun timus pada 20 hari p.i. Kemampuan benalu teh menghambat kerusakan oksidatif yang disebabkan oleh radikal bebas berkaitan dengan aktivitas bahan aktif pada benalu teh sebagai antioksidan. Daun dan batang tanaman ini mengandung alkaloid, flavonoid, glikosida, triterpen, saponin, dan tanin yang berperan sebagai antioksidan (Windardi dan Rahajoe, 1998; Achi, 2005). Ekstrak benalu teh spesies Scurrula atropurpurea mengandung 16 bahan bioaktif yang terdiri dari enam senyawa asam lemak, dua santin, dua glikosida flavonol, satu glikosida monoterpen, satu glikosida lignan, dan empat flavon (Ohashi et al., 2003). Uji aktivitas antioksidan menggunakan radikal bebas 1,1-difenil-2-pikrilhidrazil (DPPH), dilakukan pada ekstrak daun benalu teh S. oortiana. Semakin rendah nilai IC 50 semakin tinggi potensi antioksidannya. Nilai IC 50 ekstrak n-heksan adalah 697,68 ug/ml, ekstrak etilasetat adalah 617,03 ug/ml, ekstrak metanol 93,59 ug/ml, dan ekstrak air adalah 121,17 ug/ml (Simanjuntak, dkk., 2004). Hasil pengukuran bobot relatif limpa pada 20 p.i. tidak menunjukkan perbedaan yang signifikan diantara keempat kelompok perlakuan (Tabel 1). Kondisi tersebut menjelaskan tidak ada pengaruh perlakuan yang diberikan terhadap bobot relatif organ atau tidak terjadi imunomodulasi terhadap limpa.

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A = diberi ekstrak S. oortiana tanpa diinfeksi MDV B = tanpa diberi ekstrak S. oortiana tanpa diinfeksi MDV C = diberi ekstrak S. oortiana dan diinfeksi MDV D = tanpa diberi ekstrak S. oortiana diinfeksi MDV

Gambar 2. Rataan bobot relatif timus pada 20 hari dan 40 hari pascainfeksi Tabel 2. Rataan bobot relatif bursa fabricius, timus, dan limpa 40 hari p.i. Perlakuan Organ Bursa Fabricius Timus Limpa A 0,0009 a 0,0003 0,0059 a 0,0016 0,0031 0,0005
a

B 0,0009 a 0,0001 0,0058 a 0,0027 0,0029 0,0004


a

C 0,0011 a 0,0002 0,0047 a 0,0001 0,0027 0,0009


a

D 0,0010 a 0,0002 0,0063 a 0,0008 0,0028 a 0,0013

a Superskrip dengan huruf yang berbeda pada baris yang sama menunjukkan perbedaan yang nyata (P<0.05). A = diberi ekstrak S. oortiana tanpa diinfeksi MDV B = tanpa diberi ekstrak S. oortiana tanpa diinfeksi MDV C = diberi ekstrak S. oortiana dan diinfeksi MDV D = tanpa diberi ekstrak S. oortiana diinfeksi MDV

Pengaruh Ekstrak Benalu pada Bobot Relatif Organ Limfoid 40 Hari Pasca Infeksi
Pada akhir masa perlakuan yaitu pada 40 p.i. bobot relatif organ bursa fabricius, timus, maupun limpa (Tabel 2) tidak berbeda di antara keempat kelompok perlakuan. Hal ini dimungkinkan sudah berakhirnya masa imunosupresi sebagai tahapan awal infeksi MDV yang bersifat transien (sementara) imunosupresi, yaitu bersifat sementara. Islam et al. (2002) dan Fadly (2000) menyatakan bahwa ayam komersial mengandung antibodi maternal MDV dan kejadian imunosupresi sebagai akibat infeksi MDV tergantung pada variabel yang diukur, efek supresi pada sistem imun dapat terjadi dari awal infeksi yaitu hari ketiga sampai dengan 35 pascainfeksi. Ekstrak benalu teh spesies S. oortiana berkhasiat sebagai imunomodulator karena mampu meningkatkan rataan bobot relatif bursa Fabricius dan bobot relatif timus. Hasil tersebut tercermin dari meningkatnya rataan bobot realatif bursa Fabricius pada kelompok ayam yang diberi ekstrak S. oortiana dibanding

kelompok yang diberi ekstrak dikombinasi dengan infeksi MDV maupun kelompok ayam yang hanya diinfeksi MDV 20 hari p.i.. Bobot relalatif timus pada kelompok ayam yang diberi ektrak S. oortiana dan diinfeksi MDV tidak mengalami penurunan pada 20 hari pascainfeksi, hal ini menunjukkan bahwa ekstrak S. oortiana mampu menghambat imunosupresi akibat infeksi MDV.

Kesimpulan
Uji tantang MDV onkogenik dengan dosis 1,0 X 103 TCID50 pada ayam ras petelur betina menimbulkan imunosupresi pada 20 hari p.i. berdasarkan ukuran bobot relatif bursa fabricius dan bobot relatif timus. Terjadi pemulihan menjadi normal pada 40 hari p.i. berdasarkan ukuran bobot relatif bursa Fabricius dan timus. Pemberian ekstrak benalu teh spesies S. oortiana dosis 10 mg/kg bb pada ayam ras petelur betina berpotensi sebagai imunomodulator ditandai dengan peningkatan bobot relatif bursa fabricius dan bobot relatif timus pada 20 hari p.i.

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Daftar Pustaka
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Ohashi, K., M. Mukai, P. Simanjuntak and P. H. Shibuya, 2003. Cancer cell invasion inhibitory effects of chemical constituents in the parasitic plant Scurrula artopurpurea (Lorantaceae). Chemical Pharmacology Bulletin 51(3): 343-345. Payne, L.N., and K. Venugopal, 2000. Neoplastic disease: Mareks disease, avian leucosis and reticuloendotheliosis. International Epizothyology 19(2): 544-554. Silva, R.F., S. Reddy and B. Lupiani, 2004. Expansion of a unique region in the mareks disease virus genome occurs concomitantly with attenuation but is not sufficient to cause attenuation. Journal of Virology 78(2): 733-740. Simanjuntak, P., T. Parwati, L.E. Lenny, S. Tamat dan R. Murwani, 2004. Isolasi dan identifikasi senyawa antioksi dan dari ekstrak benalu teh, Scurrula oortiana (Korth) danser (Lorantaceae). Jurnal Ilmu Kefarmasian Indonesia 2(1): 6-9. SPSS Inc., 1999. SPSS for Windows: Base Systems Users's Guide Release 10.0. Michigan Avenue, Chicago. Steel, R.G.D., dan J.H. Torrie, 1991. Prinsip dan Prosedur Statistika, Suatu Pendekatan Biometrik. B. Sumantri (penerjemah). Terjemahan dari: Principles and Procedures of Statistics, A Biometrical Approach. Gramedia Pustaka Utama. Jakarta. Tambunan, R., M. Bustanussalam, P. Simanjuntak dan R. Murwani, 2003. Isolasi dan identifikasi kafein dalam ekstrak air daun benalu teh, Scurrula junghuni, Lorantaceae. Jurnal Ilmu Kefarmasian Indonesia 1(2): 16-18. Winarno, H., K. Ohashi, M. Mukai, P. Simanjuntak dan H. Shibuya, 2003. Uji Bioaktivitas terhadap Invasi Sel Kanker dari Beberapa Senyawaan Flavonoid, Santin, Terpen, dan Ligan yang Diisolasi dari Benalu teh (Scurrulla atropurpurea) Lorantaceae. Proseding Seminar dan Pameran Nasional Tumbuhan Obat Indonesia XXIV. Pusat Studi Biofarmaka LP-IPB Darmaga, Bogor 19 20 September 2003. Hlm.141-149. Windardi, F.I., dan J.S. Rahajoe, 1988. Keanekaragaman Benalu Teh di Pulau Jawa. Warta Tumbuhan Obat Indonesia 4: 25-29. Young, J.F., C.L. Steffensen, J.H. Neilsen, S.K. Jensen and J. Stagsted, 2002. Chicken model for studying dietary antioxidant reveal that apple (Coxs orange)/Broccoli (Brassica oleracea L. var. italica) stabilizes erythrocytes and reduces oxidation of insoluble muscle proteins and lipids in cooked liver. Journal of Agriculture and Food Chemistry 50: 5058-5062.

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Vol. 9 No.3

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Vol. 9 No.3

Efektivitas Pemberian Ekstrak Temulawak (Curcumae xanthoriza) dan Kunyit (Curcumae domestica) dan sebagai Immunostimulator Flu Burung pada Ayam Niaga Pedaging
(Effectiveness of Temulawak (Curcuma xanthoriza) and Kunyit (Curcumae domestica) Extracts to Enhance Productivity and as Immunostimulator of Avian Influenza in Broiler)
Sufiriyanto dan Mohandas Indradji
Fakultas Peternakan Universitas Jenderal Soedirman, Purwokerto

ABSTRACT: The objective of the experiment was to investigate the effectiveness of treating broiler with temulawak (Curcuma xanthoriza) and kunyit (Curcumae domestica) extracts to enhance productivity and as immunostimulator of avian influenza. Broilers were given either temulawak, kunyit or temulawak+kunyit extracts. The treatments, including a control, were arranged in a factorial design. Variables measured were production index and immune titter with haemaglutination inhibition (HI) test at 35 days of age. Results showed that control, temulawak-, kunyit- and temulawak+kunyit-treated chicken have production indexes of 302.80, 382.30, 327.71, and 358.30, respectively. HI test results were all negative. It can be concluded that neither temulawak, kunyit nor temulawak+kunyit extracts is effective immunostimulator of avian influenza in broiler. Nevertheless, temulawak-treated chicken showed highest production index. Key Words: Avian influenza, haemaglutination inhibition, temulawak, kunyit

Pendahuluan
Penyakit Flu burung (Avian Influenza) terjadi outbreak (wabah) di beberapa daerah, disebabkan oleh virus H5N1 yang terjangkit di beberapa daerah Jawa Tengah, Jawa Barat dan Jawa Timur pada tahun 2003. Selanjutnya timbul wabah kedua tahun 2004-2005, daerah Sulawesi Utara (Gorontalo), di Sulawesi Selatan (Makasar) yang menyerang populasi ayam petelur, di pulau Jawa (Sukabumi, Cirebon, Boyolali dan Tegal). Pengendalian penyakit pada tahun 2003 melalui program vaksinasi dianggap cukup berhasil mereda terjangkitnya penyakit Flu burung. Tingkat keberhasilan vaksinasi berdasarkan uji titer antibodi Hemaglutinasi Inhibisi (HI), sampel darah diambil 3 minggu setelah vaksinasi, dengan nilai titer HI minimal 16 (2 4). Hasil dilapangan sangat bervariasi tergantung banyak faktor yang mempengaruhi titer antibodi tersebut, diantaranya faktor manajemen. Adapun faktor manajemen yang dapat dilakukan peternak adalah mempersiapkan ayam sebelum vaksinasi agar mencapai tingkat kekebalan optimal dengan melalui pemberian vitamin atau obat-obatan tradisional (herbal medicine). Pemberian temulawak dan kunyit dapat meningkatkan kekebalan tubuh karena kandungan

fitokimia kurkumin temulawak adalah desmetoksikurkumin dan bisdesmetoksikurkumin, fitokimia kunyit adalah desmetoksikurkumin. Zat fitokimia inilah yang berfungsi untuk meningkatkan nafsu makan, meningkatkan sekresi empedu, memperbaiki fungsi hati dan memperbaiki tampilan limfosit darah. Apabila ayam sehat dan kebal dari penyakit maka nilai produktivitasnya menjadi optimal. Untuk menilai produktivitas ayam niaga pedaging digunakan standard nilai indeks produksi, semakin tinggi nilai indeks produksi maka semakin baik cara pemeliharaannya. Perhitungan indeks produksi ditentukan oleh besaran pertambahan bobot badan harian (daily gains), angka kematian (mortality) dan nilai konversi pakan (Feed Convertion Ratio, FCR). Penelitian bertujuan untuk meningkatkan produktivitas ayam niaga pedaging berdasarkan nilai Indeks Produksi (IP), dan mengetahui kemampuan sifat immunostimulator terhadap titer kekebalan Flu burung berdasarkan uji Hemaglutinasi Inhibisi (HI test).

Metode Penelitian
Materi Penelitian
Penelitian dilaksanakan menggunakan ayam niaga pedaging sebanyak 60 ekor, pakan starter 60 kg, pakan finisher 150 kg, vitamin, vaksin ND,

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vaksin Gumboro, vaksin Flu burung (AI), tempat pakan dan tempat minum 20 set dan petak kandang 20 unit, ekstrak temulawak dan ekstrak kunyit.

Rancangan Penelitian
Penelitian eksperimental telah dilaksanakan menggunakan pola faktorial dengan Rancangan Acak Lengkap (RAL). Perlakuan P 0 sebagai kontrol, P 1 pemberian ekstrak temulawak, P 2 pemberian ekstrak kunyit dan P3 pemberian temulawak dicampur kunyit, setiap unit kandang berisi 3 ekor dan diulang sebanyak 5 kali. Peubah yang diamati adalah indeks produksi dan titer antibodi penyakit Flu burung (uji titer HI test) pada ayam niaga pedaging umur 35 hari. Pengambilan sampel darah untuk uji titer kekebalan, diambil melalui vena Brachialis sebanyak 2 ml per ekor pada umur 35 hari (pemeriksaan hematologis), untuk satu unit diambil satu ekor (Siregar, 1988). Data dianalisis dengan analisis ragam, dan apabila terdapat perbedaan dilanjutkan dengan uji BNJ (Steel dan Torrie, 1980).

Hasil dan Pembahasan


Produktivitas Ayam Pedaging
Produktivitas ayam niaga pedaging diukur berdasarkan nilai Indeks Produksi (IP). IP adalah perbandingan antara pertambahan bobot badan harian (g) dikalikan daya hidup (100% - persentase mortalitas) dibagi konversi pakan (FCR) dikalikan sepuluh (Chapmann, 1988). IP hasil penelitian ini menunjukkan non signifikan yang berarti pemberian ekstrak temulawak dan kunyit memberikan efek produktivitas sama dengan kontrol atau pemberian vitamin dan antibiotika, dengan kata lain bahwa temulawak dan kunyit dapat digunakan untuk mengganti antibiotika dan vitamin pada pemeliharaan ayam niaga pedaging mulai umur 21 hari sampai dengan umur 35 hari. IP hasil penelitian ini menunjukkan perbedaan secara biologis, P 0 kontrol indeks produksi sebesar 303,98; P 1 perlakuan temulawak 382,06; P 2 perlakuan kunyit 327,51 dan P3 perlakuan campuran temulawak dan kunyit sebesar 358,26. Hal ini diduga pemberian temulawak dan kunyit mampu membunuh kuman patogen Escherichia coli (E. coli) dalam saluran pencernaan (Hadi, 1985) sehingga kuman non patogen tumbuh menjadi optimal, karena kandungan kurkuminoid dan minyak atsiri (Purnomowati dan Yoganingrum, 1981) bersifat membunuh kuman E. coli dan kuman

patogen lain dalam usus, sesuai dengan Sufiriyanto (1998) yang menyatakan bahwa pemberian probiotik (Lactobacillus sp) dapat membunuh kuman E. coli sebesar 80% pada ayam niaga pedaging sehingga dapat meningkatkan bobot badan (pada umur 6 minggu dari kontrol 1723 g menjadi 1868 g), menurunkan konversi pakan, meningkatkan protein efisiensi dan meningkatkan indeks produksi dari 229 menjadi 290. Pada penelitian ini, pemberian temulawak dosis 0,5 g per liter memberikan IP sebesar 382,06 dan hasil ini dikategorikan berhasil baik sesuai dengan Chapmann (1988) yang menyatakan bahwa IP merupakan indikator pemeliharaan ayam niaga pedaging (kategori kurang baik bila IP dibawah 200, kategori baik bila IP 200-250, dan kategori baik sekali bila IP 250-300, serta sangat baik sekali bila IP di atas 300). Fadilah dan Polana (2004) menyatakan bahwa IP ayam niaga pedaging dikatakan baik apabila mempunyai nilai diatas 200, semakin tinggi nilai indeks produksi menunjukkan pemeliharaannya semakin baik. Hasil indeks produksi ini dipengaruhi bobot badan, pada penelitian ini bobot badan umur 35 hari mencapai 1.824,46 g dengan perlakuan pemberian temulawak. Hal ini lebih baik dibandingkan dengan standard North dan Bell (1990) yang menyatakan ayam umur 5 minggu bobot badan 1460 g dan umur 6 minggu mencapai bobot badan 1890 g, sedangkan Pauzenga (1990) mengatakan bahwa bobot badan 1800- 2000 g dicapai pada ayam niaga pedaging umur 40-42 hari. Secara umum pertumbuhan ayam pedaging akan berkembang sesuai dengan perkembangan kemajuan teknologi pakan dan genetik sehingga dari waktu ke waktu hasil pencapaian bobot badan akan berubah sesuai dengan kualitas, kondisi dan situasi setempat. Konversi pakan pada penelitian menunjukkan tidak berbeda nyata secara statistik (P>0,05), tetapi secara biologis menunjukkan perbedaan yaitu pada P 0 (kontrol) konversi pakan sebesar 2,09, P 1 sebesar 1,91; P 2 sebesar 2,03 dan P 3 sebesar 1,97. Angka konversi pakan semakin kecil menunjukkan hasil yang optimal ditunjukkan pada perlakuan P 1 atau perlakuan pemberian ekstrak temulawak sebesar 0,5 g per liter air minum. Hal ini sesuai dengan Guritno (2002) menyatakan pemberian temulawak dapat menurunkan konversi pakan sehingga secara otomatis dapat meningkatkan indeks produksi dari 290,52 menjadi 302. Kenaikan bobot badan harian ayam niaga pedaging mulai umur 21 sampai 35 hari pada

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penelitian ini menunjukkan P 0 (kontrol) sebesar 61,60 g, P 1 (temulawak) sebesar 72,78 g, P 2 (kunyit) sebesar 64,30 g dan P 3 (temulawak dan kunyit) sebesar 70,09 g. Pemberian ekstrak temulawak dengan dosis sebesar 0,5 g per liter air minum memberikan hasil penambahan bobot badan harian yang paling optimal dibandingkan dengan perlakuan yang lain. Hal ini sesuai dengan pendapat Nataamijaya et al. (2000) bahwa pemberian kunyit pada ayam pedaging mampu meningkatkan bobot badan dari kontrol 1,37 kg menjadi 1,55 kg dengan pemberian jamu kunyit dan lempuyang perlakuan K5L4 atau kunyit 0,04% dan lempuyang 0,02% diracik dalam pakan ayam pedaging diberikan selama 5 minggu. Peningkatan pertambahan bobot badan juga terjadi pada kelinci yang diberi temulawak dalam pakan pada level 0,80% (Haryanto, 2006). Kandungan kurkuminoid meningkatkan kecernaan pakan (Guritno, 2001), bersifat laktagoga (Achyas dan Rasydah, 2005) dan apabila kunyit level 0,04% dalam ransum dicampur dengan lempuyang level 0,16% dapat meningkatkan bobot badan dan menurunkan angka kematian pada ayam niaga pedaging (Nataamijaya et al., 2000). Anang dan Ihsan et al. (2000) melaporkan bahwa pemberian temulawak dan kunyit dapat meningkatkan kekebalan tubuh dan menyembuhkan penyakit hepatitis.

Tingkat Kekebalan Titer HI pada AI


Pada waktu ayam niaga pedaging berumur 21 hari dilaksanakan vaksinasi flu burung sebanyak 0,5 ml per ekor secara injeksi subcutan, pelaksanaan pengambilan darah pada umur 35 hari. Hasil penelitian menunjukkan titer nol atau dapat dikatakan tidak adanya kekebalan berdasarkan uji Hemaglutinasi Inhibisi (HI test), kemungkinan lain adalah faktor dari kualitas vaksin (Nurhandayani, 2004; Kawaoka et al.,1987; Kodihalli et al., 1994), faktor strain virus (Dharmayanti et al., 2005b; Kamaludin, 2006), faktor individual ayam (Harimoto dan Kawaoka, 2001; Dharmayanti et al., 2005a; Sufiriyanto dan Indradji, 2005) dan faktor kurang pekanya metode titer HI test sehingga diperlukan uji lebih canggih seperti uji PCR (Polymerase Chain Reaction). Walaupun hasil titer kekebalan AI nol atau negatif tetapi ayam masih mampu hidup, hal ini kemungkinan di dalam tubuh telah terjadi proses kekebalan yang bersifat seluler (Abbas et al., 1991; Rantam et al., 2004), tetapi menurut Aamir et al. (2005) bahwa titer nol sangat rentan terhadap penyakit karena ayam dapat

dikatakan mampu melindungi uji tantang AI minimal skor 10 sedangkan titer HI dikatakan mampu melindungi ternak ayam apabila uji titer kekebalan HI menunjukkan Geometrik HI 15 atau 2 4 . Titer kekebalan yang baik apabila lebih besar atau sama dengan 2 4 sesuai dengan Priyono (2004), Nurhandayani (2004) yang mengatakan bahwa titer antibodi ayam sehabis di vaksin dianggap berhasil apabila nilainya lebih besar atau sama dengan 2 4 dan kisaran tersebut dianggap mampu melindungi ternak ayam dari serangan penyakit AI (Swayne et al., 2000; Tabbu, 2000; Setijanto, 2005). Pengambilan sampel darah pada tiga minggu setelah vaksinasi AI, kemungkinan hasil tersebut kurang optimal sebab untuk titer HI sebaiknya dilaksanakan satu bulan sampai dua bulan setelah vaksinasi (Hofstad et al., 1978; Kristina et al., 2004). Menurut Wood et al. (1985) waktu empat minggu ini diperlukan tubuh untuk mengadakan reaksi antigen (vaksin) dengan immunoglobulin sehingga terbentuk antibodi (Akoso, 1993). Tingkat kekebalan atau antibodi menunjukkan kemampuan tubuh untuk proteksi terhadap agen infeksi (Alexander et al., 1986; Abbas et al., 1991). Pemeriksaan ini penting untuk penelitian lapangan pada tempat-tempat individu yang divaksinasi dan yang belum pernah divaksinasi yang dipilih secara acak (Barus, 2004). Kemampuan vaksin tidak ditentukan oleh perangsangan terjadinya antibodi serum saja tetapi lebih dipengaruhi adanya penambahan proteksi terhadap penyakit (Bellanti, 1993). Menurut Tizzard (1983) bahwa tanggap kebal atau sensitifitas ternak dapat ditentukan dengan menemukan antibodi khusus didalam serum darah karena hewan atau ternak terpapar atau terinfeksi antigen tertentu. Ayam pedaging yang tidak divaksin AI kemungkinan besar melindungi diri dari serangan penyakit melalui mekanisme resistensi nonimunologis. Faktor-faktor yang berperan antara lain adalah lisozim, empedu dan hati, sumsum tulang, kelenjar timus dan yang utama adalah faktor interferensi dan interferon. Mekanisme pertahanan antiviral non-imunologis interferensi adalah istilah nama penghambatan replikasi virus karena adanya virus lain, karena virus lain tersebut menghasilkan interferon (Kimball, 1994) dan interferon dilepaskan sel yang terinfeksi atau tertulari virus dalam beberapa jam setelah invasi virus maka interferon sudah terproduksi dalam jumlah yang banyak (Tizzard, 1983). Interferon terbentuk apabila terjadi infeksi virus yang pertama atau penyakit baru

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muncul dan yang terbentuk adalah interferon tipe 1 (IFN type I) yang berfungsi menghambat proses replikasi virus dan biasanya bersamaan dengan kerja NK (Natural Killer cell) yang berfungsi melisiskan sel target infeksi (Abbas et al ., 1991). Sistem kekebalan ayam niaga pedaging yang sakit AI maka pada awal infeksi tubuh membentuk kekebalan melalui peningkatan sitokin sedangkan sitokin sendiri dalam tubuh macamnya banyak sekali sehingga diantara sitokin bersifat antagonis, mengakibatkan gagal pernafasan atau pneumonia akut. Pemberian temulawak dan kunyit mampu menekan sitokin, sehingga secara tidak langsung dapat menekan kejadian AI pada ayam. Menurut Nidom (2005) bahwa pemberian temulawak dapat menekan jumlah sitokin dan menghambat perkembangan virus saat virus mengalami perbanyakan diri (replication). Kandungan zat fitokimiawi temulawak dan kunyit berfungsi memperbaiki fungsi hati atau berfungsi hepatoprotektor (Dalimartha, 2000ab) dan dari tanaman obat bekerjasama memperkuat sel terhadap serangan virus pada berbagai lini mulai dari mencegah penetrasi, mencegah multiplikasi sampai dengan mencegah keluarnya virus dari dalam sel, lebih baik lagi apabila mengekstrak temulawak dan kunyit menggunakan air panas (Mursito, 2001). Selain efek menghambat replikasi virus, temulawak dapat berfungsi sebagai immunostimulator fagositosis dan meningkatkan kemampuan limfosit (Dalimartha, 2000a), hepato stimulan (Liang et al., 1985) dan hepatoprotektor karena mencegah kerusakan sel hati sehingga proses metabolisme dapat berlangsung lancar (Harmanto, 2007). Hal ini sesuai dengan pendapat Endrini (2007) bahwa flu burung dapat ditanggulangi dengan minum tanaman obat tradisional yang bersifat antivirus dan bersifat immunostimulan serta tanaman obat yang memiliki efek konstruktif yaitu mampu memperbaiki jaringan dan kelenjar yang rusak.

Daftar Pustaka
Aamir, G., N. Shaamoon, Y. Mohammed and N. Jawad, 2005. Immunomodullatory effects of multistrain probiotics (Protexin) on broiler chicken vaccinated againts Avian Influenza Virus (H9). International Journal of Poultry Science 4(10): 777-780. Abbas, A.K., A.H. Lichtman and Y.S. Pober, 1991. Cellular and Molecular Immunology. WB. Saunders Company. Philadelphia. Pp. 4-6, 38-45, 309-310. Achyat, D.E., dan R. Rasyidah, 2005. Kunyit (Curcumae domestica Val). http//www.asiamaya.com/jamu/isi/ kunyitcurcumaedomestica.htm. (10 September 2007). Akoso, B.T., 1993. Manual Kesehatan Unggas. Kanisius. Yogyakarta, Hlm. 93-94. Alexander, D.J., G. Parsons and R.S. Manvell, 1986. Experimental assesment of the pathogenicity of eight avian influensa a virusses of H5 sub type for chickens Turkeys, duck and quail. Avian Pathol 15: 647 662. Anang, S.F.R., dan M.M. Ihsan, 2000. Temulawak dan kunyit sembuhkan hepatitis. PT. Jamu Iboe. Dalam: http/www.jamuiboe.com.artikel 04php. (10 September 2007). Barus, R.A., 2004. Kronologi Wabah Avian Influenza (AI) di Indonesia. Warta Kesehatan Hewan. Media Informasi Direktorat Kesehatan Hewan. JanuariApril 2004. Bellanti, J.A., 1993. Immunology III. Gajah Mada University Press. Yogyakarta. Chapmann, J.J., 1988. Probiotics. Accidifers and Yeast Culture a Plate for Natural Additives in Pig and Poultry Production. Biotrchnology in the Feed Indistries. Proceedings of Alltechs for Fourth Annual Symposium.Pp.219-223. Dalimartha, S., 2000a. Atlas Tumbuhan Obat Indonesia. Trubus Agriwidya, Jakarta. Dalimartha, S., 2000b. Tiga Puluh Enam Resep Tumbuhan Obat untuk Menurunkan Kolesterol. Panebar Swadaya, Jakarta. Dharmayanti, N.L.P.I, R. Indriani, R. Damayanti, A. Wiyono dan R.M.A. Adjid, 2005. Karakter virus avian influensa isolat Indonesia pada wabah gelombang ke dua. Jurnal Ilmu Ternak dan Veteriner 10(3) : 217-226.

Kesimpulan
Indeks Produksi optimal pada penelitian ini adalah 382,30 yang diperoleh pada pemberian ekstrak temulawak dosis 0,5 g per liter air minum. Pemberian kunyit dosis 0,25 g per liter air minum menghasilkan Indeks Produksi sebesar 327,80, dan pada campuran temulawak dan kunyit menghasilkan Indeks Produksi sebesar 358,30. Titer HI pada AI tidak menunjukkan adanya perbedaan kekebalan antara perlakuan dengan kontrol.

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Mikrobiologi Indonesia, 27-28 Agustus 2004. Hlm. 41. Universitas Diponegoro. Semarang. Retno, F.D. dan J. Suryani, 1998. Penyakit-penyakit Penting pada Ayam. Edisi ke 4. Medion. Bandung. Setijanto, H., 2005. Avian Influenza: Epidemiologi Penyakit dan Strategi Pencegahannya. Diskusi Pelaksanaan Penelitian, 20 Desember 2005. Dikti. Jakarta. Steel, R.G.D. and J.H. Torrie, 1980. Principles and Procedure of Statistics. 2 rd ed., Mc. Graw Hill., International Book. Co., Singapura. Subbarao, K., H. Chen, D. Swayne, L. Mingay, E. Fodor, G. Bromnlee., X. Xu. X. Lu, J. Katz, N. Cox, and Y. Matsuoka, 2003. Evaluation of a genetically modified reasortant H5N1 influenza a virus vaccine candidate generated by plsmid based reverse genetics. Virology 305: 192-200. Swayne, D.E., M. Garcia, J.R Beck, N. Kinney, and D.I. Suarez, 2000. Protection against diverse higly pathogenic H5 avian influenza viruses in chickensimmunized with a recombinant fowlpox vaccine containing an H5 avian influenza hemagglutinin gene insert. Vaccine 18: 1088-1095. Tabbu, C.R., 2000. Penyakit Ayam dan Penanggulangannya. Penyakit Bakterial Mikal dan Viral. Volume 1. Penerbit Kanisius Yogyakarta Hlm. 233- 245. Tizzard, I., 1983. Pengantar Immunologi Veteriner. Airlangga University Press. Surabaya. Hlm.143275. Wood, J.M, Y. Kawaoka, L.A. Newberry, E. Bordwell, and R.G. Webster, 1985. Standardization of inactivated H5N2 influenza vaccine and efficacy against lethal A. Avian Diseases 29: 68-78. immunized with a recombinant fowlpox vaccine containing an H5 avian influenza hemagglutinin gene insert. Vaccine 18: 1088-1095. Tabbu, C.R., 2000. Penyakit Ayam dan Penanggulangannya. Penyakit Bakterial Mikal dan Viral. Volume 1. Penerbit Kanisius Yogyakarta Hlm. 233- 245. Tizzard, I., 1983. Pengantar Immunologi Veteriner. Airlangga University Press. Surabaya. Hlm.143275. Underwood, J.C.E., 2000. Patologi Umum dan Sistematik. Penerbit Buku Kedokteran EGC. Jakarta. Wood, J.M, Y. Kawaoka, L.A. Newberry, E. Bordwell, and R.G. Webster, 1985. Standardization of inactivated H5N2 influenza vaccine and efficacy against lethal A. Avian Diseases 29: 68-78.

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Vol. 9 No.3

Comparative Analysis of the Level of Social Integration in Cattle Farmers Groups in Banjarnegara District, Indonesia
(Analisis Perbandingan Tingkat Integrasi Sosial pada Kelompok Peternak Sapi Potong di Kabupaten Banjarnegara, Indonesia)
Mochamad Sugiarto* and Yusmi Nur Wakhidati
Faculty of Animal Science, University of Jenderal Soedirman, Purwokerto

ABSTRAK: Pembentukan dan pengembangan kelompok peternak akan dapat meningkatkan kekompakan dan kebersamaan antar peternak dalam melakukan aktifitas usaha peternakan. Melalui kelompok peternak, anggota akan bersatu dan bekerja sama untuk menjadikan mereka lebih berdaya (empowered). Integrasi sosial (social integration) di dalam kelompok peternak merupakan faktor yang penting dalam upaya peningkatan proses pemberdayaan anggota dan keberlanjutan kelompok ternak. Penelitian ini bertujuan untuk mengetahui dan membandingkan tingkat integrasi sosial peternak dalam kelompok peternak yang digolongkan berdasarkan tingkat perkembangan organisasi (organizational development) di Kabupaten Banjarnegara. Ketiga golongan kelompok peternak tersebut adalah pemula (beginner), madya (intermediate) dan lanjut (advance). Delapan kelompok peternak yang terdiri dari 145 peternak dipilih sebagai responden dengan menggunakan metode pengambilan sample berjenjang. Data tentang tingkat integrasi sosial dikumpulkan menggunakan kuisioner terbuka dan tertutup. Uji Kruskal-Wallis digunakan untuk membandingkan tingkat integrasi sosial pada tiga golongan kelompok ternak berdasarkan tingkat perkembangan oganisasi. Hasil penelitian menunjukkan bahwa integrasi sosial peternak di dalam kelompok peternak di Kabupaten Banjarnegara sudah cukup tinggi. Berdasarkan uji Kruskal-Wallis, tingkat integrasi sosial peternak menunjukkan perbedaan (P<0,05) pada ketiga golongan kelompok ternak yang ada di Kabupaten Banjarnegara (pemula, madya dan lanjut). Peternak pada kelompok peternak golongan lanjut (advance) mempunyai rata-rata tingkat sosial integrasi yang relatif lebih tinggi dibandingkan pada kelompok peternak golongan pemula (beginner) dan madya (intermediate). Kata Kunci: Pemberdayaan, integrasi sosial, tingkat perkembangan organisasi

Introduction
The development of the livestock sector in Indonesia has generated a number of achievements and contributions such as increasing job opportunity, poverty reduction, and increasing quantity and quality of animal production (Rohaeni and Hamdan, 2004). However, the small-scale farmers still remain a major player in national cattle development. These small scale farmers are characterized by (1) limited number of cattle owned by farmers, (2) using simple technology, (3) the cattle farming is just a sideline business, and (4) labor intensive. These characteristics have resulted in low bargaining power of farmers and high sensitivity to change (Yusdja et al., 2001). Hence, it is necessary that farmers be encouraged to increase productivity through technical, economic, and institutional enhancement.
*

Corresponding author. Tel.:+62 281 638792, Fax.:+62 281 626080, e-mail: zoegic@yahoo.com

The effort to increase cattle population technically and institutionally in Banjarnegara has been done simultaneously through improving the farmers knowledge and practice in cattle production. Establishment of cattle farmers group in the village and sub district level, which has been supervised by Local Government Unit (LGU) of Banjarnegara District since 1999 was aimed at increasing the cattle farmers welfare. The group approach in empowering farmers would increase unity and thereby collective action. Through the association, farmers can learn from each other and foster mutual cooperation among themselves. Besides, through a farmers group, members unite and cooperate which become a strategic action for making them powerful. The lifespan and success of farmers group depends on the unity and active involvement of members in the groups activities. This could ensure survival of the organization to benefit all members. Diversity has recently captured the attention of those interested in group performance. Group members can differ in functional specialization and

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demographic or cultural identities, such as age, race, sex, and citizenship (Chatman and Flynn, 2001). However, differences among group members give rise to varied ideas, perspectives, knowledge, and skills that can improve their ability to solve problems and accomplish their work by providing suitable environment for members to integrate into a group (Ely and Thomas, 2001). Integration of an individual into a group will in the long term improve socio-economic conditions and participatory opportunities for individuals. It will benefit society as a whole through the achievement of stability, economic security and social cohesion. A highly cohesive group would promote empowerment. Cohesiveness increases the members willingness to listen to and be influenced by group mates. On the other hand, inability of farmers to adjust in a group will lead to low cohesiveness and high turnover rate from a group. Therefore, social integration is required to promote members empowerment, group sustainability, and achievement of goals (OReilly, 1989). Social integration is key predictor of group performance and empowerment (Smith et al., 1994). However, integrating farmers into farmers groups are not really easy. Based on report from the Regional Livestock Office of Banjarnegara (2004) there were 56 groups of farmers who were engaged in cattle farming. However, in 2003, this number was significantly reduced to only 16 farmers groups which were classified into beginner, intermediate and advance level of farmers group. It indicates the difficulty of integrating farmers into groups and maintaining them as a group. Interpersonal conflict among members is inevitable which could result to diminished cooperation, generate dissatisfaction among members and finally lead to group disintegration. Under the well-developed farmers group, the members would be able and capable to perform their duties and roles as farmers. However, there are three different levels of farmers group in Banjarnegara based on organizational development. The differences of group development level of farmers groups in Banjarnegara would determine the level of social integration of farmers in a group. Therefore, there is an urgent need to take a deeper look into the level of social integration of cattle farmers groups in Banjarnegara District in three different level of organizational development of farmers groups.

In general, this study aimed to analyze the level of social integration in cattle farmers groups in Banjarnegara District, Indonesia. Specifically, it aimed to (1) Determine the level of social integration in cattle farmers groups in Banjarnegara District, and (2) Compare the level of social integration among three different levels of organizational development of the farmers groups.

Research Methods
The survey research was used to gather information from respondents of cattle farmers groups in Banjarnegara. The population of the study comprised of the 16 cattle farmers groups in Banjarnegara Regency. These cattle farmers groups are supervised by the Livestock Service of the Local Government Unit of Banjarnegara. The multi-stage sampling method was used in this survey research. The multi-stage sampling involves selecting a sample in at least two stages. In the first stage, the researcher stratified large groups into three specific strata of farmers groups (beginner, intermediate, and advance) and then, the researcher selected randomly farmers groups in each stratum. In the second stage, within a farmers group, response was obtained on a variety of questions for selected individual farmers in a group by simple random sampling which was taken proportionally based on the number of members. Face-to-face interviews were conducted with the 145 respondents of cattle farmers groups which were taken proportionally from the eight selected farmers groups in each stratum comprised the sample based on Slovin Formula. This formula was used to improve accuracy of sample from large number of population (Agustina, 2003). Observation values of those groups were at ordinal data. KruskalWallis test was used to compare the level of social integration of three different level of farmers group development.

Results and Discussion


Aspects of Social Integration
Social integration refers to the degree to which group members are attracted to the group, feel satisfied with other members, interact socially with them, and feel psychologically linked to one another (Smith et al., 1994). The level of social integration into the group is reflected in the behavioral patterns

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of group members. It is a substantive incorporation of all members to build a shared set of norms, values, and culture for a new group. Table 1 present the aspects of social integration as measurement of the level of social integration. It was measured through (a) individually perceived attraction to the group, (b) satisfaction with other members, (c) social interaction among members, (d) interpersonal conflict, (e) interpersonal cooperation, and (f) motivation to sustain relationships. Attraction to the group. It was indicated that majority (77.24%) of the respondents had been attracted much to the farmers group and only a few (1.38%) of them were not attracted much by the group. The table shows that most of the farmers desired to be identified and accepted as member of the farmers group due to some reasons. It was found that most members of intermediate and advanced farmers groups had slightly higher attraction to the group (82.86% and 80.43%, respectively) than members in beginner farmers groups (71.88%). Members who are attracted strongly with their group should cooperate more with group interests and exert greater effort on behalf of the group (Kramer, 1991). Satisfaction with other members. More than two-thirds (67.59%) of the respondents expressed a feeling of satisfaction after some periods of experience within the farmers group. As defined, satisfaction is related to the degree of adequacy to meet the needs, feeling with other members, and experience to have relationship with other members. Members in the intermediate farmers group (74.29%) expressed more satisfaction to have relationship with other members than farmers in the beginner (60.94%) and advanced (71.74%) farmers groups. The members in intermediate farmers group have high social sensitivity. It indicates that the members has high ability to understand a member better than others in farmers group indicated by respect, empathy, help, and care about each other in group activities. Besides, group communication in intermediate farmers group tends to be better than other farmers groups. These facts cause the members of the intermediate farmers groups to have high satisfaction to stay with other members. Smith (1994) explained that members of groups with better communication process experience higher morale and satisfaction and, most importantly, exhibit greater efficiency in the coordination of tasks

Group interaction. Majority of the respondents (78.62%) had moderate interaction with other members in a group and only a few (2.07%) had low interaction with other members in group. Overall, cattle farmers in advanced farmers group tend to have more interaction with other members. The more dynamic the group is and the clearer rules are giving more space for members to interact with others. Dovidio et al. (1998) interpersonal social interactions in a group are able to achieve productive negotiations, conflict resolution and personal empowerment. Interpersonal conflict. It was found that all the respondents did not experience conflict with other members of farmers group in terms of incompatible goals, scarce resources, and involvement of other parties in achieving goals. All (100%) of the respondents cited that they seldom experienced conflict and easily solved these conflicts during meeting. The results show that the advanced farmers group had lesser conflict compared to other levels of farmers groups. This suggests that farmers had achieved solidarity in their group as one of the goals in the establishment of the farmers group. Jehn and Mannix (2001) supported that higher development stage of a group has generated low conflict among members. Interpersonal cooperation. Majority (51.03%) of the respondents exhibited moderate cooperation with others while only very few (0.69%) of them did not. It reveals that the respondents tend to work together, share, collaborate, and cooperate with other members to achieve goals and run the daily activities in cattle production smoothly. The findings show that members in the advanced farmers groups cooperated more frequently with other members to achieve farmers group goals. Interpersonal cooperation was done more often in advanced farmers group along with more interactions. Van Dyne et al. (1995) stated that higher stage of group development with high cohesiveness shall enhance members' desires to help one another. Motivation to sustain relationship. Majority (70.34%) of the respondents expressed high motivation to sustain relationship with other members in the group. No farmer (0%) reported poor motivation to sustain that relationship. Members in the intermediate and advanced farmers groups tended to have higher motivation to sustain relationship with others. Hence, a higher stage of

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group development would result in satisfaction of members to stay longer in organization. Ugboro (2006) explained that people in more developed group tend to have higher job satisfaction and therefore they would gain higher loyalty to stay in a group.

Level of Social Integration in Three Different Levels of Organizational Development of Farmers Groups
There is an increasing awareness that integration will improve socio-economic conditions and participatory opportunities for individuals, as well as benefits society as a whole, through the achievement of stability, economic security and social cohesion (Rudiger, 2003). Participation is high among members of group when they are really

empowered. To empower the people, unity and integration of people in a group must be guided and formed strongly. Highly cohesive groups would promote empowerment. Social integration is really imperative for the promotion of individual, group, community, and societal well-being (Nelson et al., 2001). Level of social integration in farmers groups is the degree to which an individual is linked to others in the group (O Reilly, 1989). In this study, the level of social integration is a summation of the scores from six subjects (attraction to the group, satisfaction with other members, social interaction among members, interpersonal conflict, interpersonal cooperation, and motivation to sustain relationships).

Table 1. Aspects of social integration as measurement of the level of social integration across three different levels of organizational development of farmers groups Level of Organizational Development Beginner Intermediate Advanced Aspects (n=64) (n = 35) ( n = 46) n % n % n % Level of Attraction to the Group High ( > 44) 46 71.88 29 82.86 37 80.43 Moderate (28-44) 17 26.56 5 14.29 9 19.57 Low (<28) 1 1.56 1 2.86 0 0.00 Mean score 45.26 46.40 46.39 Level of Satisfaction with Other Members High ( > 18) 39 60.94 26 74.29 33 71.74 Moderate (12-18) 25 39.06 9 25.71 13 28.26 Low (< 12) 0 0.00 0 0.00 0 0.00 Mean score 18.36 20.14 19.26 Frequency of Group Interaction Often (>18) 11 17.19 5 14.29 12 26.09 Moderate (12-18) 52 81.25 29 82.86 33 71.74 Seldom (<12) 1 1.56 1 2.86 1 2.17 Mean score 16.69 16.54 16.80 Occurrence of Interpersonal Conflict Often (>26) 0 0.00 0 0.00 0 0.00 Moderate (17-26) 0 0.00 0 0.00 0 0.00 Seldom (<17) 64 100.00 35 100.00 46 100.00 Mean score 7.75 7.43 8.08 Interpersonal Cooperation Often (>21) 30 46.88 13 37.14 27 58.70 Moderate (14-21) 33 51.56 22 62.86 19 41.30 Seldom (<14) 1 1.56 0 0.00 0 0.00 Mean score 20.89 20.66 21.59 Motivation to Sustain Relationship High (>26) 39 60.94 30 85.71 33 71.74 Moderate (17-26) 25 39.06 5 14.29 13 28.26 Poor (<17) 0 0.00 0 0.00 0 0.00 Mean score 26.86 27.46 27.09 All Groups (n = 145) n 112 31 2 % 77.24 21.38 1.38 45.89 67.59 32.41 0.00 19.07 19.31 78.62 2.07 16.69 0.00 0.00 100.00 8.01 48.28 51.03 0.69 21.06 70.34 29.66 0.00 27.07

98 47 0

28 114 3

0 0 145

70 74 1

102 43 0

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Table 2. Test of difference on the level of social integration among three different levels of organizational development of farmers groups in Banjarnegara Level of Organizational Development Beginner Intermediate Advanced (n=64) (n = 35) ( n = 46) n % n % n % 1 51 12 1.56 79.69 18.75 135.81a 102 -145 7 28 0 20 80 0 138.62 b 131 -151 10 35 1 21.74 76.09 2.17 139.93 c 130 -150 All Groups (n = 145) n 17 115 13 % 11.72 79.31 8.97 137.80 102 151

Aspects Level of Social Integration High ( > 144.85) Medium (130.40 144.85) Low (< 130.40) Mean score Interval
a,b,c,

Means in the same row with different superscripts are significantly different (P<0.05)

This study found that majority (79.31%) of the respondents had medium level of social integration and only few (8.97) of the respondents had low level of social integration. Table 2 shows that members of the advanced farmers groups have relatively higher level of social integration. The Kruskal-Wallis analysis showed that there is significant difference (P<0.05) of level of social integration among the three different levels of farmers groups. Based on Table 2, it can be observed that farmers groups at different level of organizational development (OD) tend to have higher level of social integration among members. This confirms the finding of Robbins (2002) where he concluded that members of highly-developed groups were more satisfied. It is expected that advance groups will be more cohesive, integrated, interact frequently, and exhibit more off-task and interpersonal behavior. This is added by Shaw (1981) that groups whose members are more socially integrated should be able to coordinate their efforts and integrate their perspectives more effectively and efficiently, yielding a coherent and timely final product.

Increasing level of social integration in cattle farmers group could improve participation, empowerment and sustainability membership of cattle farmers. 2. The results show that three levels of organizational development of farmers groups existed although the level of social integration among them differed. Advanced level farmers group had the highest level of social integration. Building and improving capacity of farmers group as an advanced group would be a great challenge to increase level of social integration.

References
Agustina, N.Z., 2003. Pengaruh kinerja tugas dan kinerja kontekstual terhadap kepuasan kerja dan komitmen afektif. Jurnal Bisnis dan Ekonomi Maret: 10-17. Chatman, J.A., and F.J. Flynn, 2001. The Influence of demographic heterogeneity on the emergence and consequences of cooperative norms. Academy of Management Journal 44: 956-974. Dovidio, J.F., M. Geoffrey and G.A. Michele, 1998. A social psychology of national and international group relations. Journal of Social Issues Winter edition : 109-127. Ely, R.J., and D.A.F. Thomas, 2001. Cultural diversity at work: the effects of diversity perspectives on work group processes and outcomes. Administrative Science Quarterly 46 : 229-273. Jehn, K.A. and E.A. Mannix, 2001. The dynamic nature of conflict: a longitudinal study of intragroup conflict and group performance. Academy of Management Journal 44 : 255-282. Kramer, R.M., 1991. Intergroup relations and organizational dilemmas: The role of categorization processes. Research in Organizational Behavior 13 : 191-227.

Conclusion
To empower the people, unity and integration of people in a group must be strongly guided and formed. Social integration in farmers groups is therefore, a precondition of empowerment of cattle farmers. The study has looked into the level of social integration and factors related to the level of social integration. Based on the objectives of study, the following conclusions were drawn : 1. It has been considered how the level of social integration becomes an important entry point to improve empowerment of farmers. The finding led to the conclusion that level of social integration in cattle farmers group in Banjarnegara was generally moderate.

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Nelson, G., L. Prilleltensky and H. Macgillivary, 2001. Building value-based partnerships: toward solidarity with oppressed groups. American Journal of Community Psychology 29 : 127-138. OReilly, C.A, D.F. Caldwell and W.P. Barnet, 1989. Work group demography, social integration and turnover. Administrative Science Quarterly March Edition : 21-37. Robbins, P.S., 2002. Organizational Behavior. Prentice Hall Pubsliher. United States. Rudiger, A., 2003. Meeting The Challenge: Equality, Diversity And Cohesion. Paper presented at the Joint European Commission/OECD Conference, Brussel, Belgium. Regional Livestock Office of Banjarnegara, 2004. The Development of Farmers Group in Banjarnegara. Local Government Unit of Banjarnegara. Rohaeni E.S., dan A. Hamdhan, 2004. Profil Dan Prospek Pengembangan Usahatani Sapi Potong Di Kalimantan Selatan. Balai Pengkajian Teknologi Pertanian Kalimantan Selatan. Shaw, M.E., 1981. Group Dynamics: The Psychology of Small Group Behavior. McGraw Hill, New York.

Smith, K.G., K.A. Smith, J.D. Olian, H. P.Jr. Sims, D.P. OBannon, and A.J. Scully, 1994. Top management team demography and process: the role of social integration and communication. Administrative Science Quarterly 39:412-438. Smith, K.G., K.A. Smith, J.D. Olian, H.P. Sims, D.P. O'Bannon, and J.A. Scully, 1994. Top management team demography and process: the role of social integration and communication. Administrative Science Quarterly 39 : 412-438 Ugboro, I., 2006. Organizational Commitment, Job Redesign, Employee Empowerment and Intent to Quit Among Survivors of Restructuring and Downsizing. Institute of Behavioral and Applied Management. Van-Dyne, L., L.L.Cummings and J. M. Parks, 1995. Extra-role behaviors: in pursuit of construct and definitional clarity (a bridge over muddled waters). Research in Organizational Behavior 17:215-285. Yusdja, Y., H. Malian, B. Winarso, R. Sayuti, dan A.S. Bagyo, 2001. Analisis Kebijaksanaan Pengembangan Agribisnis Komoditas Unggulan Peternakan. Laporan Hasil Penelitian. Laporan Penelitian. Pusat Penelitian dan Pengembangan Sosial Ekonomi Pertanian, Bogor.

ANIMAL PRODUCTION, September 2007, hlm. 190 196 ISSN 1411 2027 Terakreditasi No.56/DIKTI/Kep/2005

Vol. 9 No.3

Why Dry Matter Intake (DMI) in Early Lactation Does Not Fullfill the Entire Energy Requirement of the Lactating Cow? : A Review
(Mengapa Intake Bahan Kering Selama Awal Laktasi Tidak Dapat Memenuhi Seluruh Kebutuhan Energi pada Sapi Perah Laktasi? : Suatu Kajian Pustaka)
Budi Rustomo *
Faculty of Animal Science, University of Jenderal Soedirman, Purwokerto

ABSTRAK: Regulasi intake merupakan fenomena biologis yang kompleks. Berbagai macam signal seperti hormon dan metabolit dapat berperan secara potensial dalam penurunan nafsu makan sapi perah setelah kelahiran. Meskipun leptin telah diusulkan sebagai salah satu faktor yang berpengaruh dalam regulasi intake, kemungkinan leptin tidak berperan besar dalam penurunan intake sapi perah selama awal laktasi. Berdasarkan asumsi strategi sapi perah untuk memobilisasi simpanan lemak tubuh selama awal laktasi, umpan balik signal yang berperan besar dalam regulasi intake selama laktasi adalah produk-produk metabolis dari pemecahan simpanan lemak tubuh. Signal yang berperan dalam penurunan nafsu makan setelah kelahiran kemungkinan berhubungan dengan peningkatan uptake metabolit dalam liver dan oksidasi asamasam lemak dari produk pemecahan lemak tubuh. Pemahan tentang kemungkinan alasan-alasan mengapa sapi perah tidak dapat meningkatkan regulasi intake pakan secara maksimum segera setelah kelahiran, dapat memberi gambaran dan arah dalam memformulasikan hipotesis-hipotesis riset tentang mekanisme penekanan intake bahan kering sapi perah selama awal laktasi. Kata Kunci: Leptin, bahan kering, energi, sapi perah

Introduction
Post partum transition period (3-4 week post partum) has been recognized as the most critical phase of the lactation cycle for the dairy cow. At this period the nutrients demands are increasing dramatically for milk production, but typically, the cows experience slow recovery of feed intake after calving until they reach their peak feed intake approximately 8-4 weeks post partum. Hence, the dry matter intake (DMI) does not fulfill the entire energy demand of the early lactating cow, which results in the cows state of negative energy balance (NEB). When the cow enters the energy-deprived state, body fat storage is mobilized. This mobilized body fat provides extra energy for the increased energy demand during early lactation. However, over mobilization of body fat can contribute to the health problems in transition dairy cow (Drackley, 1999). The objectives of the present paper are to evaluate why dairy cow does not up regulate maximum feed intake shortly post-partum partum, and how is the possible mechanism involved in the slow increase of DMI during post-partum early lactating period.

Why Does Cow Mobilize Body Fat Reserves Post Partum?


It is usually assumed that the body fat mobilization during early lactation is entirely a response to a shortfall in food energy intake relative to milk energy output. This implies that increasing energy content of the food being offered would decrease body fat mobilization in early lactation, or decreasing the nutrient availability would result in an increase in mobilization of body fat (e.g. Friggens et al., 1998). However, there are many studies show that this is not always the case (eg. Grummer et al., 1995). In fact, the use of body reserves to meet their nutrient requirement during lactation is extraordinary in several species such as in seal and whales (Oftedal, 1993), and in rat (Friggens et al., 1993). Even in situations where the environmental pressure to mobilize body lipid in early lactation is removed, for instance by providing abundant levels of nutritional resources, why should the mother still mobilize body reserves? Mother nature has accorded the ability of the mammals to produce milk for survival of the newborn. The mammary glands have a high metabolic rate during lactation, but its products do not give direct effect to the mother. This indicates that mammals have the ability to give a high priority to lactation in order to ensure the survival of their

Corresponding author: e-mail: brustomo@voguelph.co

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young (Bauman, 2000). It is proposed that body reserves mobilization in early lactation and the subsequent gain in body reserves during pregnancy are to a large extent genetically driven (Friggens et al., 2004), and body fat mobilization is not a response but rather a natural component of a safeguarding reproductive success by strategic use of body reserves (Knight, 2001, and Friggens, 2003). Lactation of the modern dairy cow may also still be viewed as a part of the cows natural roles to maintain their reproductive function. In this view, the ability of the dam to support demands of future offspring in a constrained environment will depend to a large extent on the size of her body reserves (Friggens, 2003). The vast majority of mammals have evolved the strategy of preparing for forthcoming lactation by accumulating body fat reserves. For example, the proportion of lactational resources derived from body mobilisation can approach 100% in some species of seals, whales and bears (Oftedal, 2000). The ability to provide the young with easily digestible nourishment independently of food availability at that time and location has been proposed to be the main evolutionary advantage of lactation (Vernon and Pond, 1997). At the start of lactation, failure to produce milk will result in the death of the calf, so at this time there is a benefit in having a large body reserves to safe guard milk production. Also, by using body reserves to produce milk, the dam spends less time foraging and stays closer to protect the young from the predators. As lactation progresses towards weaning the negative consequences of lactational failure and the risk of predator attack diminish, the need for large body reserves reduce. Thus, the body fat mobilisation during early lactation of dairy cow should not only be viewed as a nutritional or energetic support for high milk production, but should also be viewed as a part of the cows strategy to achieve the goal of maintaining reproductive cycle. In this perspective, the slower feed intake recovery after parturition, may not solely be viewed as the cows inability to maximize intake, but also be associated with the cows natural ability to optimize the survival of the young.

Does Leptin Play A Role in Coordinating the Feed Intake in Early Lactation?
Voluntary dry matter intake (VDMI) in ruminants is negatively correlated with body

reserves at a given physiological state (Bines and Morant, 1983). Increased fatness or body reserves also have been indicated to down regulate VDMI in sheep (Foot, 1972). This supports the lipostatic theory (Kennedy, 1953) that the regulation of body reserves and food intake is coordinated. Parabiosis studies in rat gave further proof that humoral signals are involved in the coordination of body fat and feed intake (e.g. Hervey, 1959, and Parasmewaran et al., 1977). An adipose-derived signal, which is now known as leptin, has been implicated to be the factor that may involve in maintaining energy reserves at a presumed set point by regulating feed intake, energy expenditure and whole body energy homeostasis in rodents and human (Karen et al., 1998). However, in periparturient ruminants, plasma leptin declined progressively before parturition and remained low during early lactation (Ingvarsten and Boisclair, 2001). Thus, in early lactating cows plasma leptin may not reflect changes in adiposity, and hence may not regulate feed intake. In contrast to most other metabolic hormones, leptin acts predominantly on regions of the brain involved in the regulation of energy metabolism, such as the arcuate, ventromedial anddorsomedial nuclei of the hypothalamus (Karen et al., 1997). Leptin is unlikely to contribute to reduce appetite at early lactation because maternal plasma leptin was progressively declined (by 50%) after parturition, and remained depressed during early lactation of sheep (Endhart et al., 2001, and Ingvarsten and Boisclair, 2001), and dairy cow (Brian et al., 2003). The decline in plasma leptin post partum has been attributed to inhibition of adipose tissue leptin synthesis associated with the negative energy balance of early lactation (Block et al., 2001). The decline in plasma leptin mirrors that of plasma insulin and glucose but is reciprocal to plasma GH and NEFA (Brian et al., 2003, and Ingvarsten and Boiscliar, 2001). Brian et al., (2003) and Block et al., 2003) found that insulin infusion is capable of increasing plasma leptin in dairy cows but this effect is attenuated considerably during early lactation. Exogenous growth hormone did not have independent effect on plasma leptin in early lactation, but it inhibits insulin mediated leptin synthesis. Therefore, in early-lactatingundernourished dairy cows, reduced plasma insulin could account for a portion of the decline in plasma leptin, but the increase in plasma GH is unlikely contribute to the post partum reduction of plasma leptin.

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The low plasma leptin post parturition in dairy cows may signal to the brain that a state of energy insufficient prevails in the periphery, because plasma leptin remained high in cows not milk after parturition (Block et al., 2001). Low plasma leptin during early lactation could have benefits to promote an increase in feed intake (Brian et al., 2001). Consistent with reduced leptin signalling, lactation is associated with increased expression of the orexigenix NPY and AgrP and a decrease in the expression of the anorexigenic POMC precursor product (Borgan et al., 2000). In fasting sheep, low plasma leptin is correlated with up regulation of NPY and AgRP, and a tendency to lower expression of POMC and CART (Adam et al., 200). Thus, low plasma leptin in the early lactation dairy cow may be related to the similar changes in neuropeptide expression. Therefore, during the early stage of lactation, regulation of feed intake may not be the most important function of leptin, and the main role of leptin post-partum may be to coordinate adaptation that promote energy conservation required to survive periods of nutritional deprivation. This is supported by the study that leptin therapy attenuates the neuroendocrines response to fasting in mice (Ahima et al., 1996). As well, reduction in plasma leptin could benefit early lactating cows by decreasing the peripheral tissue to insulin, and partition of glucose to mammary gland (Bell, 1997). In rodents, total absence of leptin is associated with insulin resistance, whereas leptin therapy stimulate peripheral glucose utilisation under basal and hyperinsulinemic condition (Barzilai et al., 1997). Additionally, low plasma leptin associated with nutritional insufficiency in early lactation is suggested to share many adaptations such as depressed reproductive and higher metabolic efficiency (Ingvarsten and Boisclair, 2001). Low plasma leptin during early lactation may signal that energy supply is not adequate for reproduction.

How Does Lipid Mobilization Down Regulate Feed Intake in Early Lactation
Voluntary dry matter intake (VDMI) in ruminants is negatively correlated with body reserves at a given physiological state (Bines and Morant, 1983). Increased fatness or body reserves also have been indicated to down regulate VDMI in sheep (Foot, 1972). This supports the lipostatic theory (Kennedy, 1953) that the regulation of body

reserves and food intake is coordinated. Parabiosis studies in rat gave further proof that humoral signals are involved in the coordination of body fat and feed intake (e.g. Hervey, 1959, and Parasmewaran et al., 1977). Elevated blood concentration of growth hormone (GH) during early lactation is believed to be the hormone responsible to increase lipolysis (Bauman, 2000). As large amount of adipose tissue are mobilized during early lactation, the plasma NEFA, glycerol and ketone bodies are increased (Drackely, 1999). Although a negative correlation were observed between feed intake and plasma levels of NEFA, glycerol and ketone bodies (Carpenter and Grassman, 1983), the mechanism is not clear. Glycerol may be one of the candidate signals for the effect of body fat and mobilization on food intake regulation during early lactation. Bray (1973) suggested glycerol as an indicator of total body fat mass. Adipose cells contain sufficient glycerol kynase to reutilize most of the free glycerol, and during hydrolysis of triglecrydes, glycerol is released from adypocytes into the plasma. Basal lipolysis and therefore glycerol release increase with fat cell size. It was hypothesized that glycerol could act as a satiety signal if the hypothalamus could sense the amount of glycerol being converted to glucose in the liver (Bray, 1973). However, Langhans et al. (1983) concluded that hypophagic effect of glycerol was not caused by glycerol or by the conversion of glycerol to glucose. Subcutaneous administration showed a variable results, and only non-physiological levels reliably reduced food intake in rats (Carpenter and Grosman, 1983). Glycerol may influence feeding through central nervous system (CNS) mechanism since intracerebroventricular (ICV) infusion of glycerol in rats has been shown to suppress feeding (Davis et al., 1981). Scharrer and Langhas (1990) stated that metabolism of glycerol appears to be essential its hypophagic effect in rats. However, elevated plasma glycerol level by infusion and feeding glycerol did not influence on feed intake (Carpenter and Grossman, 1983), and the authors concluded that plasma glycerol levels are relatively minor lipostatic influence on hunger. As well, Ingvartsen et al. (1999) reported that in early lactation higher glycerol only found in heifers but not found in cows that were on high feeding level pre-partum. Thus, glycerol does not seem to be a strong candidate for a signal mediating lower intake in periparturient cattle.

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An increased rate of lipolysis and oxidation of fatty acids, in the early lactation cows, is usually associated with an increased production of ketones bodies (Drackely, 1999). Hence, ketone bodies may be one of the satiety factors depressing food intake. Subcutaneous injection of beta hydroxyl butyrate (BHBA) has shown to cause hypophagia in rats (Fisler et al., 1995), and it seemed likely that the liver is involved in depression of intake by during injection of BHBA (Sharrer and Langhans, 1990). However, Stricker et al. (1977) argued that the liver does not involved in the satiety effect of BHBA because liver can not synthesize BHBA beyond acetoacetate. Sun et al. (1997) found that ICV infusion of BHBA reduced body weight but not food intake. The effect of BHBA on intake regulation during the early lactation may not be strong. NEFA is another possible candidate for feedback signals for feed intake regulation during the early lactation. Plasma NEFA concentration is elevated during early lactation, and there is a negative relationship between the plasma NEFA and VDMI in cattle (Grummer, 1993). Continuous long term intravenous infusion of long chain free fatty acids has been shown to reduce DMI in rats (Carpenter and Grossmann, 1983). Ingvarsten and Andersen (2000) suggested that fatty acids oxidation in the brain and liver may provide possible signal in intake regulation during early lactation. Kasser et al. (1985) reported that rates of fatty acids oxidation in the ventrolateral hypothalamus (VLH) and fatty acids synthesis in ventromedial hypothalamus (VMH) were linearly related to the peripheral energy status. The VLH of underfed rats had 40% increased in fatty acid oxidation and 9% decrease of glucose oxidation, and the overfed rats had a 36% decrease in fatty acid oxidation and 20% increase in glucose oxidation, compared to the control. They proposed that if an increase in free fatty acids oxidation for undernourished rats inhibits activity of VLH cells that are normally activated by glucose, then it may be that the elevated plasma free fatty acids concentration may lead to a decrease in feed intake. This is because activity of VLH increases feed intake and activity of VMH decreases feed intake (Harris and Martin, 1984). However, Beverly and Martin (1991) has failed to test this hypothesis that inducing chronic changes (14d) in VLH fatty acids oxidations did not affect feed intake. Thus, it is unlikely that fatty acids oxidation in the VLH plays an important role in regulating feed intake.

Liver of dairy cows after parturition is faced with markedly increased uptake of NEFA mobilized from adipose tissue. Carnitine palmitoyl-transferase1 (CPT-1) an enzyme involved in the uptake of NEFA into the hepatic mitrochondria has been suggested to have a significant effect on determining the relationship between fatty acids oxidation and feed intake (Drackley, 1999). Studies have shown that CPT-1 activity is greater in 1 21 days post partum than at 65 d postpartum (Dann et al., 2000). CPT-1 is inhibited by malonyl CoA, which is formed from acetyl-CoA, by acetyl-CoA carboxylase. Acetyl-CoA carboxylase is active during the well-fed state, when insulin is high, and conversely it is inactive during undernourished state, when insulin is low (Drackley et al., 2001). Malonyl-CoA concentration is responsive to changes in insulin and glucagons in ruminants (Knapp and Baldwin, 1990), increasing when insulin increase and vice versa. In rodents models, decrease in the sensitivity of CPT-1 to inhibition by malonylCoA follow decrease in insulin, which serve to amplify the signal and increase transport of NEFA into mitochondria (Zammit, 1996). Thus, during early lactation, when plasma insulin is relatively low, the increased CPT-1 concentration (Dann, 2000) may be due to a decrease in malonyl-CoA concentration or due to decrease sensitivity of CPT1 to malonyl-CoA inhibition. When the activity of CPT-1 is increased, uptake and oxidation of NEFA by the hepatic mitochondria is increased. Total oxidation of palmitate by liver homogenates was about 12% greater at day1 postpartum than at 21day prepartum (Drackley et al., 2001). Studies on the effects of free fatty acids oxidation on feed intake have been done by using mercaptoacetate, which depress Acyl-CoA dehydrogenase activity (Sharer and Langhans, 1986) or using methyl palmoxirate, which depresses the mitrochondrial CPT-1 concentration (Friedman et al., 1990a). Inhibiting fatty acids oxidation with administration of methyl palmoxirate increased feed intake in rats, particularly when the rate of fatty acids oxidation is relatively high (Friedman et al., 1990b). Langhans and Scharrer (1987) concluded that the hypophagia is most likely mediated by hepatic receptors, because inhibition of fatty acid oxidation by mercaptoacetate diet was partially blocked by hepatic branch vagotomy. Scharrer and Langhans (1988) proposed that the hypophagia is most likely linked to mitochondrial oxidation of NEFA, which give satiety signals to the brain

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mediated by vagal afferents. Thus, the satiety signals may be initiated, at least in part, by hepatic receptors, which then stimulate the vagal afferents in order to communicate signals of metabolic status to the brain (Ingvarsten et al., 1999). Additionally, Langhans (1996) proposed that satiety could be resulted from fatty acids oxidation in mitochondria that may affect the hepatocyte membrane potential through cytosolic ATP and Na-pump activity. This is supported by study of Friedman (1999) showing that methyl palmoxirate-induced increase in feed intake were negatively correlated with liver ATP content, and ATP to ADP ratio, indicating that a decrease in fatty acid oxidation can stimulate feed intake by reducing hepatic energy production.

Metabolism, Growth and Reproduction. P.B. Cronje, ed. CAB International, Wallingford, UK. Berille, N. and Faverdin, P., 1996. Lipid metabolism and intake behaviour of dairy cows : effects of intravenous lipid and beta-adrenegic supplementation. Journal of Dairy Science 79:1209-1220. Beverly, J.L., and R.J. Martin. 1991. Influence of fatty acid oxidation in lateral hypothalamus on food intake and body composition. American Journal of Physiology 261:R339-R343. Bines, J.A., and S.V. Morant.1983. The effect of body condition on metabolic changes associated with intake of food by the cow. British Journal of Nutrition 50:8189. Bray, G.A., 1973. Peripheral metabolic factors in the regulation of feeding. In: Appetite and food intake, Silverston, T Berlin Ed : Abakon Verlagsgessellschaft. Pp.141-176. Carpenter, R.G., and S.P. Grossman, 1983. Plasma fat metabolites and hunger. Physiology and Behaviour 30 : 57-63 Dann, H.M., G.N. Douglas, T.R. Overton, and J.K. Drakcley, 2000. Carnitine palmitoutransferas activity in liver of periparturient dairy cows. Journal of Dairy Science 83(Suppl.1): 251.(Abstr.). Davis, J.D., D. Wirtshafer, K.E. Asin, and D.Brief, 1981. Sustained intracerebroventricular infusions of brain fuels reduces body weight and food intake in rats. Science 212:81-82. Drackley, J.K., 1999. Biology of dairy cows during the transition period: the final frontier. Journal of Dairy Science. 82 : 2259-2273. Drackely, J.K., T.R. Overton, and G.N. Douglas. 2001. Adaptation of glucose and long-chain fatty acid metabolism in liver of dairy cows during the periparturient period. Journal of Dairy Science 84 : E100-E112. Endhart, R.A., R.M. Slepetis, A.W. Bell, and Y.R. Boisclair, 2001. Martenal leptin is elevated during pregnancy in sheep. Domestic Animal Endocrinology 21:85-96 Fisler, J.S., M. Egawa, and G.A. Bray, 1995. Peripheral 3-hydroxybutyrate and food intake in a model of dietary-fat induced obesity: effect of vagotomy. Physiology and Behaviour 58:1-7. Foot, J.Z., 1972. A note on the effect of body condition on the voluntary food intake of dried grass wafers by Scotish black face ewes. Animal Production 14:131-134.

Conclusion
Based on the cows strategy to mobilize body fat during the early lactation, the metabolic products from lipolysis are likely the possible candidates for signalling the regulation of feed intake. Feedback signals from the elevated oxidation of NEFA in the liver possibly contribute to the down regulation of feed intake during the early lactation when mobilisation of body fat is high. The low plasma insulin concentration during early lactation may result in elevating CPT-1 activity due to a decrease in malonyl-CoA concentration and due to a decrease in sensitivity of CPT-1 to malonyl-CoA inhibition. When the activity of CPT-1 is increased, uptake and oxidation of NEFA by the hepatic mitochondria is increased. This may initiate satiety signals to hepatic membrane potential, and then it stimulates the vagal afferents in order to communicate signals to the satiety center of the hypothalamus. These mechanisms may result in slow recovery of feed intake after calving, and hence DMI in early lactation does not fulfil the entire energy requirement of the lactating cow.

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