Ruminal fermentation, production and fatty acid quality of milk of late lactation dairy goat fed pufa-diet supplemented with Yeast and C. xanthorrhiza Roxb.

RUMINAL FERMENTATION, PRODUCTION AND FATTY ACID QUALITY OF MILK OF LATE LACTATION DAIRY GOAT FED PUFA-DIET

SUPPLEMENTED WITH YEAST AND Curcuma xanthorrhiza Roxb

ENDANG SULISTYOWATI

THE GRADUATE SCHOOL

BOGOR AGRICULTURAL UNIVERSITY BOGOR

PERNYATAAN MENGENAI DISERTASI DAN

SUMBER INFORMASI SERTA PELIMPAHAN HAK CIPTA*

Dengan ini saya menyatakan bahwa disertasi berjudul Ruminal Fermentation, Production and Fatty Acid Quality of Milk of Late Lactation Dairy Goat Fed PUFA-Diet Supplemented with Yeast and C. xanthorrhiza Roxb adalah benar karya saya dengan arahan dari komisi pembimbing dan belum diajukan dalam bentuk apa pun kepada perguruan tinggi mana pun. Sumber informasi yang berasal atau dikutip dari karya yang diterbitkan maupun tidak diterbitkan dari penulis lain telah disebutkan dalam teks dan dicantumkan dalam Daftar Pustaka di bagian akhir disertasi ini.

Dengan ini saya melimpahkan hak cipta dari karya tulis saya kepada Institut Pertanian Bogor.

Bogor, Februari 2014

Endang Sulistyowati

RINGKASAN

ENDANG SULISTYOWATI. Fermentasi Rumen, Produksi dan Kualitas

Asam Lemak Susu pada Kambing Perah dengan Fase Laktasi Akhir yang Diberi Diet –PUFA yang Disuplementasi Ragi dan C. xanthorrhiza Roxb

Dibimbing oleh TOTO TOHARMAT, ASEP SUDARMAN dan KOMANG G.

WIRYAWAN.

RINGKASAN

Dalam rangka memenuhi kebutuhan susu nasional dan pada saat yang sama memperbaiki kualitas susu, dalam hal ini asam lemak dari ternak ruminan laktasi seperti kambing perah, diperlukan perbaikan nutrisional dalam ransum. Hasil- hasil penelitian sebelumnya telah dilakukan pada kambing atau sapi perah yang diberi suplemen ragi, pasta tape- temulawak atau curcuma (C. xanthorrhiza

Roxb), Tabut blok, dan konsentrat laktasi yang mengandung ragi dan temulawak dapat meningkatkan produksi susu dan memperbaiki kualitas susu. Oleh karena itu, tiga penelitian yang meliputi evaluasi nutrisi konsentrat-PUFA (polyunsaturated fatty acid) selama penyimpanan, analisis ransum-PUFA secara fermentasi in vitro, dan aplikasi ransum- PUFA secara in vivo telah dilakukan. Ransum yang mengandung sumber PUFA, yaitu jagung giling sangrai, tepung kedelai sangrai, dan minyak jagung yang disuplementasi ragi Saccharomyces cereviseae dan curcuma diaplikasikan pada kambing perah dengan fase laktasi akhir.

Penelitian pertama bertujuan untuk mengevaluasi konsentrat- PUFA dengan suplementasi ragi dan curcuma yang disimpan selama 2- 6 minggu. Perlakuan konsentrat- PUFA yaitu tanpa suplemen ragi dan curcuma (PC0), 0.5% ragi (PCY), 2% curcuma (PCC), dan 0.5% ragi dan 2% curcuma (PCM). Ragi (mengandung 3.6 x 107 cfu/g) dan curcuma (mengandung 0.8% curcumin dan 1.58% tannin) disiapkan sendiri. Hasil menunjukkan bahwa kandungan kadar air dan S. cereviseae (20.68 x 106 cfu/g) meningkat nyata (P<0.05) pada penyimpanan 6 minggu; sedangkan, bahan kering (BK), bahan organik (BO), serat kasar (SK), dan bahan ekstrak tanpa nitrogen (Beta-N) menurun nyata (P<0.05). Total PUFA (P), P/S, mono unsaturated fatty acid (MUFA), dan long chain fatty acid (LCFA) lebih tinggi pada konsentrat- PUFA dengan 2% curcuma (PCC). Sebaliknya, pada konsentrat- PUFA dengan 0.5% ragi dan 2% curcuma (PCM) tinggi lemak tak jenuh dan rasio U/S. Secara keseluruhan, digabung dengan level nutrisi lainnya, pada penyimpanan 2- 6 minggu, konsentrat- PUFA dengan kombinasi 0.5% ragi dan 2% curcuma (PCM) adalah yang paling bisa dipertimbangkan secara nutrisi dan kesehatan.

Penelitian kedua, in vitro, dilakukan untuk mengevaluasi performa fermentasi rumen dari ransum-PUFA (mengandung konsentrat-PUFA, limbah kedelai dari industry tahu dan rumput Gajah) yang disuplementasi ragi dan C. xanthorrhiza Roxb. Rancangan percobaan yang diaplikasikan adalah Rancangan Kelompok dengan 4 perlakuan dengan 4 cairan rumen dari empat ekor kambing yang dipotong dan 3 ulangan masing-masing. Perlakuan yang diberikan adalah tanpa suplemen (PD0); 0.5% ragi (PDY); 2% curcuma (PDC); dan kombinasi 0.5% ragi dan 2% curcuma (PDM ). Variabel yang dianalisis adalah pH,

kecernaan bahan kering (KCBK) dan kecernaan bahan organik (KCBO), N-NH3, VFA total dan parsial, TPC, dan CH4. Hasil menunjukkan bahwa, walaupun KCBK- KCBO dan N- NH3 terendah; namun, didukung dengan populasi protozoa yang rendah dan produksi VFA yang tinggi, ransum-PUFA dengan ragi dan curcuma bisa dipertimbangkan sebagai ransum yang paling potential dalam meningkatkan metabolism nutrien dalam rumen.

Penelitian ketiga, in vivo, dilakukan untuk mengevaluasi performan ransum-PUFA dengan suplementasi ragi dan curcuma terhadap produksi dan komposisi nutrisi serta asam lemak susu, mastitis status, kecernaan nutrien, dan metabolit darah pada 20 ekor Peranakan Ettawa (PE) pada fase laktasi akhir (sekitar 4 bulan laktasi). Rancangan perlakuan berdasarkan pengelompokan tingkat produksi susu dengan menggunakan Rancangan Kelompok dengan 5 perlakuan dan 4 ulangan. Perlakuan terdiri atas tanpa aditif (PD0), 3 tablet laktasi Asifit (PDA), 0.5% yeast (PDY), 2% curcuma (PDC), dan kombinasi 0.5% yeast dan 2% curcuma (PDM) yang ditambahkan sebagai topping menjelang pemberian makan pagi (06.00) dan sore (15.00); Asifit diberikan per oral pagi hari. Sumber PUFA selain konsentrat- PUFA dari penelitian sebelumnya, juga dicampurkan kulit kedele dari limbah produksi tahu. Produksi susu dicatat sebelum, selama, dan setelah perlakuan. Perlakuan PDC mengandung lemak lebih rendah dibanding PDM, dan lebih tinggi serat kasar tetapi rendah ADF dan Phosphor (P). Pada ransum- PUFA dengan ragi (PDY) menunjukkan rendah protein. Produksi susu selama perlakuan secara kuantitatif tinggi pada PDA dan PDY. Pada pasca perlakuan, produksi ini lebih tinggi (P<0.05) dibanding perlakuan PDY dan PDC; sedangkan PDM berada diantaranya. Komposisi susu pada PDM secara kuantitas lebih rendah untuk lemak, 4% FCM, % dan berat protein, rasio lemak: protein, bahan kering, dan bahan kering tanpa lemak (BKTL), namun lebih tinggi untuk laktosa dan Ca. Indikator mastitis dan metabolit darah tidak terpengaruh (P>0.05) oleh perlakuan. Namun, secara kuantitas PDM menujukkan angka terendah untuk SCC, terendah Hb, PCV,dan glukosa, prolaktin dan trigliserida. Pada perlakuan PDM, konsumsi nutrien lemak antarperlakuan tertinggi (P<0.05), sementara, kecernaan nutrisi sebagian besar juga dipengaruhi oleh perlakuan (P<0.05 atau P<0.01). Perlakuan ransum-PUFA dengan curcuma (PDC) menunjukkan paling rendah untuk BK, BO, PK, LK, dan GE (gross energy); sementara, PDY dan PDM tinggi dalam kecernaan nutrisi. Disimpulkan bahwa 0.5% ragi dan 2% curcuma bubuk yang ditambahkan pada ransum-PUFA (PDM) adalah yang terbaik dalam hal nutrisi dan metabolit serta produksi susu pasca perlakuan (dengan semakin berlanjutnya fase laktasi) pada kambing perah.

Hasil penelitian yang berkaitan dengan asam lemak menunjukkan bahwa PDM mengandung tinggi asam lemak total, asam lemak rantai sedang, rantai panjang, dan asam lemak tak jenuh; namun, rendah dalam SCFA, n6/n3 rasio, dan indeks atherogenik. Kualitas susu kambing perah yang diberi pakan PUFA- diet dengan ragi dan curcuma ini dapat dipertimbangkan sebagai produk yang lebih sehat. Oleh karena itu, ransum PUFA dengan 0.5% ragi dan 2% curcuma merupakan ransum yang dipilih untuk diaplikasikan pada ternak kambing perah.

Kata kunci: C. xanthorrhiza Roxb, in vitro, in vivo, kambing perah, penyimpanan, ragi, ransum- PUFA.

ENDANG SULISTYOWATI. Ruminal Fermentation, Production and Fatty Acid Quality of Milk of Late Lactation Dairy Goat Fed PUFA-Diet Supplemented with Yeast and C. xanthorrhiza _{Roxb. Supervised by TOTO}

TOHARMAT, ASEP SUDARMAN and KOMANG G. WIRYAWAN.

SUMMARY

In order to meet milk demand nationally and at the same time to improve milk quality, in this case fatty acid of lactating ruminants such as dairy goat; it is required a nutritional improvement in the ration. Based on previous research results that have been applied to either dairy goat or dairy cows with supplementations of yeast, fermented cassava- curcuma (C. xanthorrhiza roxb), Tabut block, and lactating concentrate were reported to increase milk yield and milk quality. Therefore, three experiments, evaluating nutrient contents during storage of the polyunsaturated fatty acid or PUFA- concentrate, analyzing fermented PUFA- diet in vitro, and applying PUFA- diet in vivo had been conducted. Diet containing PUFA- concentrate (PUFA sources consisted of roasted ground corn, roasted soybean meal, and corn oil) supplemented with yeast and curcuma had been applied in late lactation dairy goats.

The first experiment was to evaluate the nutritive values of PUFA- concentrate supplemented with yeast and curcuma stored in 2-6 weeks. There were four PUFA- concentrates, namely, no supplement (PC0), 0.5% yeast (PCY), 2% curcuma powder (PCC), and a mixture of 0.5% yeast with 2% curcuma (PCM). Yeast (containing 3.6 x 107 cfu/g) and curcuma powder (containing 0.8% curcumin) were self made. These concentrates were evaluated for nutrition and fatty acid contents during 2 and 6 weeks of storage. Results showed that moisture and Saccharomyces cereviseae (20.68 x 106 cfu/g) increased significantly (P<0.05) in 6 weeks of storage; whereas, dry matter (DM) and organic matter (OM), crude fiber (CF), nitrogen free extract (NFE) decreased significantly (P<0.05). The total PUFA (P), P/S, monounsaturated fatty acid (MUFA), and long chain fatty acid (LCFA) contents were found higher in PUFA- concentrate with 2% curcuma powder. Whereas, the PUFA- concentrate with 0.5% yeast and 2% curcuma powder was higher in unsaturated (U) fat and the ratio of U/S. Based on all nutrient values during the storage of 2- 6 weeks, the PUFA- concentrate with 0.5% yeast and 2% curcuma powder was considered nutritionally reasonable.

The second experiment, in vitro, was conducted to evaluate the ruminal performances of PUFA-diet (containing PUFA-concentrate with soybean byproduct and King grass) supplemented with yeast and curcuma. Experimental design applied was Randomized Complete Block of 4 ruminal liquor derived from four slaughtered goats and 4 treatments (PD0, PDY, PDC, PDM ), in 3 replications. Variables analyzed were pH, N-NH3, total and partial VFA, TPC, and CH4. Results showed that in spite of having the lowest organic and dry matter digestibilities as well as N- NH3; however, supported by low protozoa

population and high VFA production in the goat rumen fluid, the PUFA- diet with a mixture of yeast and curcuma additives was considered the most potential diet to improve nutrient metabolism in rumen.

The third experiment, in vivo, was to evaluate milk production and health of dairy goat fed PUFA-diet supplemented with yeast of curcuma. The were 20 crossbred Ettawa goats in the late lactation selected based on their production levels and grouped them in Randomized Block design to receive 5 treatments of no supplement (PD0), 3 tablets of Asifit (PDA), 0.5%/d yeast (PDY), 2%/d curcuma (PDC), and a mix of 0.5%/d yeast and 2%/d curcuma (PDM) in diets containing concentrate with PUFA sources (roasted ground corn, roasted soy bean meal, and corn oil), soybean by-product, and King Grass. The variables evaluated were milk yield, milk composition, mastitis status, nutrient digestibility, and blood metabolites. Milk yield was recorded daily pre, during, and post treatment. The PDC had lower ether extract than that of mix diet and showed higher crude fiber but lower ADF; diet with yeast had lower crude protein; and mix diet showed lower P content. Milk yield during treatment was higher in PDA and PDY. Post treatment, these milk yields were higher (P<0.05) than those in PDY and PDC; while the PDM was in between. There were tendencies that mix diet (PDM) had lower milk fat, 4% FCM, % protein and weight, fat: protein ratio, dry matter, and solid non fat, but it showed higher lactose and Ca percentage. Mastitis indicators and blood metabolites were not affected by treatments. However, PDM showed lowest SCC, higher Hb, PCV, and glucose but had lower prolactin and triglyceride. In PDM, nutrient intake of ether extract was the highest (P<0.05), while nutrient digestibilities were mostly affected significantly (P<0.05 or P<0.05). The PDC had the lowest DM, OM, CP, EE, and GE; whereas PDY and PDM showed higher digestibilities. As conclusion, a mix supplement of 0.5%/d dried yeast and 2%/d curcuma powder was considered reasonable since it showed a better recovery in milk yield after treatment with progressing lactation in dairy goat.

Results dealing with fatty acid demonstrated some consistencies in diet containing polyunsaturated fatty acid (PUFA) supplemented with 0.5% yeast and 2% curcuma (C. xanthorrhiza Roxb) that were high in total fatty acid, MCFA, LCFA, and PUFA. Milk fatty acid of goat fed with this diet showed high in LCFA and MUFA; while it was low in SCFA, n6/n3 ratio, and atherogenicity index. These milk qualities were optimally considered good in terms of healthier product. Therefore, the PUFA- diet with 0.5% yeast and 2% curcuma was a reasonable choice to be applied for dairy goat.

Key words: C. xanthorrhiza Roxb, dairy goat, in vitro, in vivo, PUFA- diet, Yeast, storage

© Hak Cipta Milik IPB, Tahun 2014

Hak Cipta Dilindungi Undang-Undang

Dilarang mengutip sebagian atau seluruh karya tulis ini tanpa mencantumkan atau menyebutkan sumbernya. Pengutipan hanya untuk kepentingan pendidikan, penelitian, penulisan karya ilmiah, penyusunan laporan, penulisan kritik, atau tinjauan suatu masalah; dan pengutipan tersebut tidak merugikan kepentingan IPB

Dilarang mengumumkan dan memperbanyak sebagian atau seluruh karya tulis ini dalam bentuk apa pun tanpa izin IPB

Dissertation

as part of requirement to fulfill PhD degree at

Nutrition and Feed Science Study Program

RUMINAL FERMENTATION, PRODUCTION AND FATTY ACID QUALITY OF MILK OF LATE LACTATION DAIRY GOAT FED PUFA-DIET

SUPPLEMENTED WITH YEAST AND C. xanthorrhiza Roxb

THE GRADUATE SCHOOL

BOGOR AGRICULTURAL UNIVERSITY BOGOR

2014

Close exam examiner: Prof Dr Ir Dewi Apri Astuti, MS Dr Despal

Open exam examiner: Prof Dr Ir Ali Khomsan, MS Prof Dr Ir Lili Warly, MAgrSc

Approved by Advisory Committee

Prof Dr Ir Toto Toharmat, MAgrSc Main Supervisor

Acknowledged by Head of Study Program

Nutrition and Feed Science

Dr Ir Dwierra Evvyernie A, MS MSc

Dean of Graduate School

Dr Ir Dahrul Syah, MScAgr Prof Dr Ir Komang G. Wiryawan

Co- Supervisor

Dr Ir Asep Sudarman, MRurSc Co- Supervisor

Name : Endang Sulistyowati

SID : D162110011

Dissertation title : Ruminal Fermentation, Production and Fatty Acid Quality of Milk of Late Lactation Dairy Goat Fed

PUFA – Diet Supplemented with Yeast and Curcuma xanthorrhiza Roxb

Date of Examination: 16 January 2014

Date of Graduation:

ACKNOWLEDGEMENT

Above all, I would humbly distinguish the Most Merciful Allah swt who led me to take this opportunity, pursuing my dream, my ultimate study. Secondly, I would like to acknowledge Directorate General of Higher Education (DGHE) of the Republic of Indonesia through BPPS for granting scholarship during the enrollment in Nutrition and Feed Science at the Graduate School of Bogor Agricultural University.

I would like to sincerely deliver my greatest gratitude to my Advisors: Prof Dr Ir Toto Toharmat, MAgrSc, Dr Ir Asep Sudarman, MRurSc, and Prof Dr Komang G Wiryawan for their advice, expertness, encouragement, and support. I would also express my appreciation to Prof Dr Dewi Apri Astuti, MS, Dr Anuraga Jayanegara, and Dr Ir Sumiati, MS MSc who had guided me for the preliminar y exams; also to Dr Ir Dwierra Evvyernie A, MSc as head of study program and Mr. Supriadi as her Assistant who have been very kindly taking care of the paper work concerning the process of study at this department. Highly recognitions were addressed to Prof Dr Ir Dewi Apri Astuti, MS and Dr Despal who had been my examiners in close exam (December 3, 2013). I would also recognize Prof Dr Ir Ali Khomsan, MS and Prof Dr Ir Lily Warly, MAgrSc who had dedicated their valuable time to serve as examiners in open exam; as well as to Prof Dr Ir Luki Abdullah MAgrSc and Prof Dr Ir Panca Dewi Manuhara Karti MS in January 16, 2014. My special gratitude was also extended to Prof Dr Ir Toto Toharmat, MAgrSc, Prof Dr Ir Suryono HS, MS and Dr Ir Erwanto, MS for giving me recommendation to apply this PhD Program at the Graduate School of Bogor Agricultural University.

My appreciations were also extended to DGHE- IPB that has selectively HIBAH STRANAS- 2013; Ms. Dian for her generous help at Fapet-IPB Dairy Nutrition Laboratory; Fapet- IPB Microbiology Laboratory; Fateta- IPB Microbiology Laboratory; Ms. Endang at PAU-IPB Proxymate Laboratory; IPB- Laboratory Terpadu; Kesmavet- FKH IPB Laboratory; IPB- Biofarmaka Laboratory; Balai Penelitian Tanaman Obat dan Rempah- Deptan; Balai Besar Pascapanen Laboratory- Deptan; Physiological Nutrition Laboratory- Deptan; and Faperta-UNIB Microbiology Laboratory; and Kimia Farma Laboratory. Special thanks also went to Mr. Syauqi who allowed me to use his facility at Cordero Dairy Goat Farm in Ciapus, Bogor during the in vivo experiment; Mr. Eko and co- workers as well as IPB Polytechnic students who had worked together effortlessly during the research at the farm.

Sincere appreciations were also addressed to all my lectures, staffs and colleagues for their sharing experience, knowledge, and friendship at Bogor Agricultural University. Special thanks were extended to Drh Ria P Santoso, MS, Dr Ir Nyoto Santoso, MS, Ir Fiastri, IMBR and FWB students, Sistanto, and University of Dehasen Bengkulu as part of my big family.

I would like to take this moment to deeply express my thankful feeling to my beloved husband Ir Sigit Mujiharjo, MSAE, my precious sons Gilang Damar Waseso, Dirgandaru Galih Waskito, and Rubi Jalu Waspodo; my late father Adi Subagio and late mother Umi Rochmana, also Ibu Siti Mutiah; my brothers, sisters and their families: Mulyadi Suhendro, SE, Erma Sulfikar, SE, Rita

Kusuma Wardani, SSos, and Bambang Satrio Nugroho, SPi who have been praying and loving me as always.

This Dissertation is dedicated for you all and my students at Animal Science Department, University of Bengkulu.

Bogor, February 2014

TABLE OF CONTENT

LIST OF TABLE LIST OF FIGURE LIST OF APPENDIX

1. INTRODUCTION 1

2. THE NUTRITIVE PERFORMANCES OF PUFA- CONCENTRATE

SUPPLEMENTED WITH YEAST AND Curcuma xanthorrhiza Roxb

STORED IN 2-6 WEEKS

Introduction 6

Material and Methods 7

Results and Discussion 11

Conclusion 20

3. IN VITRO GOAT FERMENTATION OF PUFA- DIET

SUPPLEMENTED WITH YEAST AND Curcuma xanthorrhiza Roxb

Introduction 21

Material and Methods 22

Results and Discussion 24

Conclusion 29

4. MILK YIELD, MILK COMPOSITION, AND BLOOD

PARAMETERS OF LATE LACTATION DAIRY GOAT FED POLYUNSATURATED FATTY ACID DIET SUPPLEMENTED WITH YEAST AND Curcuma xanthorrhiza Roxb

Introduction 30

Material and Methods 31

Results and Discussion 35

Conclusion 43

5. FATTY ACID QUALITY OF MILK OF LATE LACTATION DAIRY GOAT FED PUFA-DIET SUPPLEMENTED WITH YEAST AND

Curcuma xanthorrhiza Roxb

Introduction 44

Material and Methods 45

Results and Discussion 48

Conclusion 54

6. GENERAL DISCUSSION 55

7. CONCLUSION 57

REFERENCES APPENDIXES BIOGRAPHY

58 68 86

LIST OF TABLE

2.1 Ingredients of PUFA-concentrate supplemented with yeast and C. xanthorrhiza Roxb

8 2.2 Fatty acid contents of corn oil, roasted soybean meal,

roasted ground corn

9 2.3 Moisture, dry matter, organic matter, and ash in

PUFA-concentrate supplemented with yeast and C. xanthorrhiza

Roxb stored for 2-6 weeks

11

2.4 Ether extract, crude protein, crude fiber, N-free extract, and gross energy in PUFA-concentrate supplemented with yeast and C. xanthorrhiza Roxb stored for 2-6 weeks

15

2.5 Neutral detergent fiber, ADF, Ca, and P in PUFA-concentrate supplemented with yeast and C. xanthorrhiza

Roxb stored for 2-6 weeks

13

2.6 Tannin and curcumin in PUFA-concentrate containing yeast and C. xanthorrhiza Roxb during 2-6 weeks storage

14 2.7 Microbes population in PUFA-concentrate supplemented

with yeast and C. xanthorrhiza Roxb stored for 2-6 weeks

15 2.8 Correlation between moisture and other nutrient in

PUFA-concentrate supplemented with yeast and C. xanthorrhiza

Roxb(PCM)

16

2.9 Fatty acid contents in PUFA-concentrate supplemented with yeast and C. xanthorrhiza Roxb stored for 2 weeks

17 2.10 Fatty acid contents in PUFA-concentrate supplemented

with yeast and C. xanthorrhiza Roxb stored for 6 weeks

19 3.1 Composition of ingredients in PUFA-diet supplemented

with yeast and C. xanthorrhiza Roxb used in in vitro goat ruminal fermentation

22

3.2 Nutrient contents of PUFA-diet supplemented with yeast and C. xanthorrhiza Roxb fermented in vitro in goat rumen liquor

24

3.3 Fatty acid contents in PUFA-diet supplemented with yeast and C. xanthorrhiza Roxb

25 3.4 In vitro dry matter and organic matter digestibility (IVDMD

and IVOMD) and fermentation characteristics of PUFA-diet supplemented with yeast and C. xanthorrhiza Roxb in goat rumen liquor

27

3.5 Protozoa, TPC, and methane gas production of PUFA-diet supplemented with yeast and C. xanthorrhiza Roxb in goat rumen liquor fermentation in vitro

28

4.1 Ingredients and nutrient composition of PUFA-diet containing Asifit, yeast and C. xanthorrhiza Roxb for dairy goat

32

4.2 _{Effects of feedingPUFA-diets supplemented with Asifit, yeast} and C. xanthorrhiza Roxb on nutrient intakes of dairy goats

4.3 _{Effects of feeding PUFA-diets supplemented with Asifit, yeast} and C.xanthorrhiza Roxb on nutrient composition of feces of dairy goats

36

4.4 _{Effects of feding PUFA-diets supplemented with Asifit, yeast and}

C.xanthorrhiza Roxb on nutrient digestibility of dairy goats

36 4.5 _{Effects of feeding PUFA-diet supplemented with Asifit, yeast and}

C.xanthorrhiza Roxb on milk yield and composition of dairy Goats

38

4.6 _{Effects of feeding PUFA-diets supplemented with Asifit, yeast} and C.xanthorrhiza Roxb on microbial composition and mastitis indicator ofdairy goats

41

4.7 _{Effects of feeding PUFA-diets supplemented with Asifit, yeast} and C.xanthorrhiza Roxb on blood parameters of dairy goats

42 5.1 Ingredients and nutrient composition of PUFA-diet containing

Asifit, yeast and C. xanthorrhiza Roxb for dairy goats

46 5.2 Nutrient composition of PUFA-diet containing Asifit, yeast and

C. xanthorrhiza Roxb for dairy goats

48 5.3 Total fatty acid contents in PUFA-diets supplemented with Asifit,

yeast and C. xanthorrhiza Roxb for dairy goat

50 5.4 Fatty acid contents in milk of dairy goat fed PUFA-diet

supplemented with Asifit, yeast and C. xanthorrhiza Roxb

51 5.5 Fatty acid contents in feces of dairy goat fed PUFA-diet

supplemented with Asifit, yeast and C. xanthorrhiza Roxb

53 5.6 Fatty acid and PUFA correlation among intake, milk, feces of

dairy goat fed PUFA-diet supplemented with Asifit, yeast and C. xanthorrhiza Roxb

54

LIST OF FIGURE

2.1 Ingredients and steps in preparing the yeast: (a) yeast ingredients; (b) mixing; (c) dough; (d) mouting-drying

7 2.2 Steps in preparing the curcuma powder: (a) shredding Curcuma

xanthorrhiza Roxb; (b) drying; (c) oven; (d) grinding

7 3.1 Some activities conducted in the in vitro experiment: (a)

PUFA-diet; (b) in vitro fermentation; (c) TPC test; (d) methane test; (e) tannin test; (f) NH3 test

23

4.1 Some activities done during the experiment: (a) PE goat; (b) additives; (c) diets; (d) blood; (e) milk; (f) feces samples

34 4.2 Milk production along the research (pre, during, and post

treatment)

LIST OF APPENDIX

1 Breed and lactation state of each dairy goat used in the research

64

2 Body measurements of dairy goat used in the research 65

3 Goat and treatment arrangement during in vivo experiment 66 4 Environment condition around housing at Cordero Farm in

Ciapus, Bogor

67

5 Anova of PUFA-concentrate moisture 70

6 DMRT of PUFA-concentrate moisture 70

7 Anova of PUFA-concentrate dry matter 70

8 DRMT of PUFA-concentrate dry matter 70

9 Anova of PUFA-concentrate ash 71

10 Anova of PUFA-concentrate organic matter 71

11 DMRT-week of PUFA-concentrate organic matter 71

12 DMRT-treatment of PUFA-concentrate organic matter 71

13 ANOVA of PUFA-concentrate ether extract 72

14 ANOVA of PUFA-concentrate crude protein 72

15 ANOVA of PUFA-concentrate crude fiber 72

16 DMRT of PUFA-concentrate crude fiber 72

17 ANOVA of PUFA-concentrate NFE 73

18 DMRT-week of PUFA-concentrate NFE 73

19 DMRT-treatment of PUFA-concentrate NFE 73

20 ANOVA of PUFA-concentrate ADF 73

21 ANOVA of PUFA-concentrate ADF 74

22 DMRT-week of PUFA-concentrate ADF 74

23 DMRT-treatment of PUFA-concentrate ADF 74

24 ANOVA of PUFA-concentrate saccharomyces 74

25 DMRT of PUFA-concentrate saccharomyces 75

26 ANOVA of PUFA-concentrate bascillus 75

27 DMRT of PUFA-concentrate bascillus 75

28 ANOVA of dry matter consumption 75

29 ANOVA of organic matter consumption 75

30 ANOVA of crude protein consumption 76

31 NOVA of crude fiber consumption 76

32 ANOVA of ether extract consumption 76

33 DMRT of ether extract consumption 76

34 ANOVA of NFE consumption 76

35 ANOVA of gross energy consumption 77

36 ANOVA of NDF consumption 77

37 DMRT of NDF consumption 77

38 ANOVA of ADF consumption 77

39 ANOVA of Ca consumption 77

40 DMRT of Ca consumption 78

41 ANOVA of P consumption 78

42 DMRT of P consumption 78

44 DMRT of dry matter digestibility 78

45 ANOVA of organic matter digestibility 79

46 DMRT of organic matter digestibility 79

47 ANOVA of crude protein digestibility 79

48 DMRT of crude protein digestibility 79

49 ANOVA of crude fiber digestibility 79

50 DMRT of crude fiber digestibility 80

51 ANOVA of ether extract digestibility 80

52 DMRT of ether extract digestibility 80

53 ANOVA of NFE digestibility 80

54 DMRT of NFE digestibility 80

55 ANOVA of gross energy digestibility 81

56 DMRT of gross energy digestibility 81

57 ANOVA of NDF digestibility 81

58 DMRT of NDF digestibility 81

59 ANOVA of ADF digestibility 81

60 DMRT of ADF digestibility 82

61 ANOVA of Ca digestibility 82

62 DMRT of Ca digestibility 82

63 ANOVA of P digestibility 82

64 DMRT of P digestibility 82

65 ANOVA of TDN 83

66 DMRT of TDN 83

67 ANOVA of milk production 83

68 ANOVA of caproic acid C6:0 in milk 83

69 DMRT of caproic acid C6:0 in milk 83

70 ANOVA of undecanoic acid C11:0 in milk 83

71 DMRT of undecanoic acid C11:0 in milk 84

72 ANOVA of lauric acid C12:0 in milk 84

73 DMRT of lauric acid C12:0in milk 84

74 ANOVA of tridecanoid acid C13:0 in milk 84

75 DMRT of tridecanoic acid C13:0 in milk 84

76 Milk production of goats fed PUFA- diets along the research

85 77 Milk production of goats in block 1 fed PUFA-diets along

the research

85 78 Milk production of goats in block 2 fed PUFA-diets along

the research

85 79 Milk production of goats in block 3 fed PUFA-diets along

the research

85 80 Milk production of goats in block 4 fed PUFA-diets along

the research

1. INTRODUCTION Background

Milk production in Indonesia is increasing yearly, however, it has not fulfilled milk demand. In 2011, milk production was 926 103 tons, while the total demand of milk and milk product was 3,903 103 tons (DGLAH 2011). The lack of this milk demand would be imported for about 76.27% a year; with an international milk price of $400/ton, it would cost around $1,190,800,000/yr. On the other hand, with 17,483 103 population of goat in 2011, supposed 0.5% out of them was dairy goat and 30% of them was in lactation with 0.5 kg/d milk production, in one lactation period would be producing milk about 1,967 tons. There might be a potential alternative of milk production as much as 0.05%/yr to supply national milk demand. This assumption might be too low compared to data released by FAO that was about 1.56% goat milk contributed by Indonesia to the world goat milk (Thiruvenkadan 2012). This huge gap of milk consumption and milk production should be narrowed. At the same time, the quality of the milk needs to be improved as well. For these purposes there has to be nutrition manipulation that could be applied to the dairy ruminants, such as dairy goat.

In terms of fatty acid (short, medium- chain, saturated branched, mono- polyunsaturated, cis- trans conjugated) contents in animal product, including milk, have been receiving a lot of attention concerning of human health (Chilliard et al. 2003). Therefore, it needs efforts in improving rumen kinetics relating to nutrients of the feed provided, especially in improving the polyunsaturated fat, PUFA. This could be achieved by applying diet rich in PUFA (such as roasted corn grain, roasted soybean meal, and corn oil) supplemented with temulawak (Curcuma xanthorrhiza Roxb) and yeast.

Curcuma xanthorrhiza Roxb, within its root, contains some bioactives such as curcuminoids (3% of dry matter) consisting of curcumin (C), demethoxycurcumin, and bisdemethoxycurcumin (Rukayadi et al. 2008); and xanthorrizol (33.2% of rhizome oil) reported by Sirat et al. (2008). Curcumin has molecular weight of C21H20O6 that would decrease due to the changes of

poliphenol weight during radiation process of curcuma powder and curcuma simplicia. This condition resulted in degradation of covalent bond that changed to free phenol becoming highly active in antioxidant compared to that of in curcumin tablet and fresh curcuma (Nurlidar and Chosdu 2008). Bioactive of curcumin also functions as antibiofilm (Rukayadi et al. 2008), antimicrobial, anti- inflammation, anticancer (apoptosis, antiangiogenesis), detoxification, neuro protection, and antiaging (Hwang 2008); elevates bile production, lessens tissue inflammation and plasma LDL (low density lipoprotein) in rabbit (Wientarsih and Meulen 2008). It was also reported that in early identification, curcumin and xanthorrhizol worked very strong as antibacterial and effectively slow down the growth of Staphylococcus aureus, Salmonella paratyphi, Trichophyton gypseum, and Mycobacterium tuberculosis (Benson 2012).

Curcumin interacts with adiposity pancreas cells, heart cells, macrophage, and muscle in suppressing proinflammatory transcription factors (Aggarwal 2010). As anti inflammation and anti pathogen, curcumin is expected to decrease inflammation in mammary gland causing mastitis. The indicators of this

are decreasing SCC (somatic cell count), Staph. aureus, and E. coli contents in milk.

In organic (less use of antibiotic synthetic) dairy farm, it was found decreasing CNS (coagulase negative staphylococci) and environment Streptococcus sp in milk sample (Suriyasataporn 2010). Normal SCC level of dairy cow milk was less than 1 x 105 cfu/ml; while the infected cow was 1 x 106 cfu/ml (BytyQi et al. 2010). It has been reported also that SCC (consisting of 75% leucocytes: neutrophils, macrophages, lymphocytes, erythrocytes, monocytes; and 25%epithelial cells) in infected dairy cows were facing potential loss of milk production for up to1200 kg/lactation with SCC of 2.2 - 4.5 x 106 cells/ml (Sharma et al. 2011). There was correlation between high level of SCC and mastitis associated with decreasing lactose, α-lactalbumin, and milk fat (Harmon 1994).

Combination of polyherbal in low level (125 mg/kg body weight) produced milk optimally in dairy goat (Mirzaei and Prasad 2011). Curcuma, including C. xanthorrhiza Roxb, also contains bioactive steroid that served as lactagogum, conserving the continuity of cell differentiation of epithel cells in normal tissue, mucose secretion, alveolus proliferation, and ductul growth in mammary gland. Hormones relating to this process such as prolactin is the most secreted hormon right before partus (Larson 1985). Concentration of prolactin in dairy goat blood decreased from 18.55 ng/ml in 15 days of lactation to 5.88 ng/ml in 150 days of lactation (Singh and Ludri 2002). Therefore, curcuma besides maintaining lactation, also containing other secondary compound, such as tannin and saponin that function in microbial rumen activity.

Ruminal kinetics are expressed by microbial rumen activity in digesting fiber that its dynamics were affected by protozoa functioning as predator for bacteria (Gutierez 2007). Saponin (triterpenoid and steroid saponin) generated from any materials were identified detrimental toward protozoa (antiprotozoa) and as defaunating agent in rumen with the effect of detergent foaming on the surface of cell membrane (Francis et al. 2002). Saponin generated from lerak (Sapindus rarak) extract of 0.8 mg/ml rumen liquor decreased protozoa population and affected composition and total bacteria population after 24 hours in vitro

fermentation. Population of F. succinogenes bacteria was dropped with increasing consentrate; whereas P. ruminicola increased markedly with increasing

Sapindus. Decreasing protozoa was associated with decreasing activity of methanogen; modifying H2 to be propionate supported by bacteria with certain

amount of population and appropriate type as well as abundant amount of substrate (Sugiarti et al. 2011).

Beside saponin, tannin is an effective poliphenol in decreasing methan (CH4) gas production that was produced about 10% out of energy , in vitro or in vivo. The efficacy of tannin depends on type of carrier materials, easily hydrolyzed (easily accumulated, toxic) or condensed (bound, safe). Tannin from chestnut was the most reasonable in reducing CH4 gas per digested organic matter

with the lowest C2/C3 ratio (Jayanegara et al. 2008). Methane gas is affected during fermentation process of fat source of diet and metabolized it through lipid hydrolysis process with end results of triglyceride, free fatty acid, glycerol, then

continued in β- oxidation to yield acetate and H2, as the precursors ofmethane

Lipid supplementation produced fatty acid, including CLA (conjugated linoleic acid) based on its source; high in cooking oil and fresh grass, but, low in unprotected grain oil (Chilliard et al. 2003). Free fatty acid and CLA in the rumen will be biohydrogenized (hydrolyzed by bacteria with addition of 2 ion H+ to be saturated; in conversely, it should be bypassed then biosynthesized as unsaturated fat in milk). Eventually, it is transported into mammary gland (as in precursor), changed from linolenic acid (C18:3 cis 9, cis 12, cis 15) in several steps to become stearate (C18:0), then ultimately converted into oleate in mammary gland. This process is optimazed by SCD (steoroyl- CoA desaturase) enzyme. Fatty acid content of milk of goat fed sunflower and soy bean oil was lowered in saturated fat and atherogenicity index (1.21- 1.71), but increased in CLA compared to control. This suggested that unsaturated fatty acid in blood vessel was going up that made this index down as expected (Chilliard et al. 2003).

Yeast, mostly mentioned as Saccharomyces cereviseae in any forms, dried or liquid, is widely used as rumen enhancer or fermenting feed supplement with variable effects on dry matter and organic matter digestibility, ruminal microbes, and ruminal fermentation activity (pH rumen, VFA, lactate, milk production, and fatty acid) as described by Desnoyers et al. (2009). Lynch and Martin (2002) reported that yeast, both in culture and cells showed no different activities; however with high dose (0.73 g/l) produced lower CH4 and C2/C3 in in vitro

fermentation of diet containing Bermuda grass. Viability of yeast cell in dry form was decreasing (from 9.88 x 1010 cfu/g to 5.43 x 1010 cfu/g) after 3 months of storage in 400C (Sullivan and Bradford 2011). Supplementation of RumiSacc, commercial yeast, was not significantly different from control in affecting BUN (blood urea nitrogen), cholesterol, triglyceride, and glucose of dairy cow (Yalcin

et al. 2011).

Synergy of temulawak (curcuma), yeast, and PUFA- concentrate has been gradually conducted in lactating ruminants. Milk production of lactating Bali cow could increase 6.6- 9.3 times higher with 5% pasta of fermented cassava and temulawak (C. xanthorrhiza Roxb) liquid, reported by (Sulistyowati 1999); milk yield has doubled with Tabut (abbreviation for fermented cassava and curcuma) block (300g) in lactating Bali cows (Sulistyowati et al. 2001); in FH dairy cow with Tabut block, its milk production increased with increasing amount (0- 450g) of the block following this model: Y: 6.66 + 0.05 x, r: 0.90 (Sulistyowati and Erwanto 2009). Microbial population of Tabut block decreased with increasing curcuma (15-25%) during storage of 3- 12 weeks (Sulistyowati et al. 2008b), while 15% curcuma level was optimal for milk production (Sulistyowati et al. 2008a) in FH dairy cows; 2%yeast supplementation in FH dairy cows produced the highest milk yield (Sulistyowati et al. 2010b); concentrate containing 10.5%C. xanthorrhiza Roxb was optimal for milk production in FH dairy cows (Sulistyowati et al. 2010a ); PUFA- concentrate containing 4.5% roasted ground corn produced the highest milk production with lower milk fat with increased PUFA and ratio of n6/n3 was about 2.14 in milk of dairy cow (Sulistyowati et al.

2010c) with high TDN (Sulistyowati et al. 2010d); curcuma concentrate in lactating dairy cow (Sulistyowati et al. 2011). Meanwhile, milk production of dairy goat supplemented with 3g/d of yeast was optimal (Sulistyowati and Mega 2002); whereas, dairy goat fed pasta of fermented cassava and curcuma liquid

could increase milk production for 0.1 kg/d with significant reduction in milk fat to 2.09% (Sulistyowati 2009).

However, along with these researches, there has not been studied on ruminal activity, blood metabolites, milk fatty acid quality and milk production, especially in dairy goat, moreover during late lactation. Therefore, three experiments had been conducted (nutrient quality of stored PUFA-concentrate, in vitro ruminal fermentation, and in vivo application of PUFA-diet supplemented with yeast and C. xanthorrhiza Roxb) in late lactating dairy goat.

Problems

Based on above description, it has been analyzed several problems that need to be improved:

1. Milk production of dairy goat was very low compared to its actual capacity.

2. Milk quality, especially milk fatty acid was high in saturated fat.

3. Ruminal fermentation needs to be manipulated with nutrient originated from PUFA –diet supplemented with curcuma and yeast.

Research Purposes

Based on the identified problems, three experiments were designed to meet those three purposes:

1. To evaluate nutritional quality of PUFA- concentrate containing roasted ground corn, roasted soybean meal, corn oil supplemented with curcuma and yeast stored in 2, 4, and 6 weeks.

2. To analyze performance of PUFA- diet supplemented with curcuma and yeast in in vitro ruminal fermentation.

3. To evaluate PUFA- diet added with Asifit tablet (a commercial tablet for women in lactation), curcuma and yeast on nutrient digestibility, blood metabolites, mammary health, production and fatty acid quality of milk of late lactating dairy goat.

Research Outcomes

Out of each experiment, there would be a result that could be applied, those are: 1. One PUFA – concentrate formula that is durable in nutritional quality

during the 2- 6 weeks of storage.

2. One PUFA- diet formula that is optimal in in vitro ruminal fermentation that is optimal metabolically.

3. One formula of PUFA- diet that is optimal for mammary health, production and fatty acid quality of milk of late lactating dairy goat.

Research Hypothesis

1. Curcumin and tannin within curcuma are able to protect nutrients of PUFA- concentrate stored for up to 6 weeks.

2. Curcumin, tannin, yeast (Saccharomyces cereviseae), and PUFA sources (roasted ground corn, roasted soybean meal, and corn oil) in PUFA- diet were able to reduce unwanted microbes, such as protozoa in such a way that would improve metabolic rumen in biohydrogenation process.

3. Curcumin, tannin, yeast (Saccharomyces cereviseae), and PUFA sources (roasted ground corn, roasted soybean meal, and corn oil) in PUFA- diet were able to improve metabolic rumen in biohydrogenation process that eventually will improve mammary health, production and fatty acid quality of milk of late lactating dairy goat.

2.THE NUTRITIVE PERFORMANCES OF PUFA- CONCENTRATE SUPPLEMENTED WITH YEAST AND Curcuma xanthorrhiza Roxb

STORED IN 2-6 WEEKS

ABSTRACT

The objective of this experiment was to evaluate the nutritive performances of PUFA- concentrate supplemented with yeast and Curcuma xanthorrhiza Roxb stored in 2-6 weeks. There were four PUFA- concentrates, namely, no supplement (PC0), 0.5%yeast (PCY), 2%curcuma powder (PCC), and 0.5%yeast with 2%curcuma powder (PCM). Yeast (containing 3.6 x 107 cfu/g) and curcuma powder (containing 0.8% curcumin) were self made. These concentrates were evaluated for nutrition and fatty acid contents during 2 and 6 weeks of storage. Results showed that moisture and Saccharomyces cereviseae (20.68x 106 cfu/g) increased significantly (p<0.05) in 6 weeks of storage; whereas, dry matter (DM) and organic matter(OM), crude fiber (CF), nitrogen free extract (NFE) decreased significantly (p<0.05). The total PUFA (P), P/S, Monounsaturated fatty acid (MUFA), and long chain fatty acid (LCFA) contents were found higher in PUFA- concentrate with 2%curcuma powder. Whereas, the PUFA- concentrate with 0.5%yeast and 2%curcuma powder was higher in unsaturated (U) fat and the ratio of U/S. Combining all nutrient performances during the storage of 2- 6 weeks, the PUFA- concentrate with 0.5%yeast and 2% curcuma powder was considered nutritionally healthy.

Key words: curcuma, nutrient, PUFA-concentrate, yeast, storage

INTRODUCTION

Concentrate is the main nutrient source for livestock, including dairy goats. Its nutritional values are importantly taken into account in order to fulfill the requirement for improved production and good quality of milk produced. Several feed supplements such as yeast have been applied extensively. It was reported as optimal level in a rate of 3g/dairy goat (Sulistyowati and Mega 2002), 20g/dairy cow (Sulistyowati et al. 2010b), 56g/dairy cow (Hristov et al. 2010), and 50g Rumisacc / dairy cow (Yalçın 2011). Yeast contain active viable cells, especially

Saccharomyces cereviseae that would be beneficial for nutrient digestion through fermentation within the concentrate all the way to rumen system.

Herbs are considered as local plants medicine and as feed supplement. Temulawak (Javanese- Indonesian) or Curcuma xanthorrhiza Roxb as other

Curcuma sp, containing curcuminoids (curcumin, demethoxycurcumin, and bisdemethoxycurcumin) known for its biological activities, such as antioxidant, anti-inflamantory, antimicrobial (Jayaprakasha et al. 2002). Supplementation of this curcuma powder of 15g/kg concentrate increased milk production of dairy cow (Sulistyowati et al. 2010a) and 60g curcuma diluent/300g block decreased total bacteria and fungi (Sulistyowati et al. 2008b). Polyunsaturated fatty acid (PUFA) supplementation in concentrate is incompletely bio hydrogenised later in the bio system of ruminants which eventually will reduce fat, short and medium chain fatty acid into long chain fatty acid in milk (Whitlock et al. 2002 and

(a) (b) (c) (d)

Figure 2.2 Steps in preparing C. xanthorrhiza Roxb powder: (a) shredding; (b) drying; (c) oven; (d) grinding

Schmidely et al. 2005). Concentrate supplemented with roasted ground corn, yeast, and curcuma powder showed optimal levels in PUFA, ratio of PUFA/saturated (P/S), and n6/n3 designated for dairy cows (Sulistyowati et al. 2010b).

Dairy goats and their products are getting popular in terms of preference and nutrition for human health. Therefore, a part of this research was designed to evaluate concentrate containing roasted corn grain, roasted soy bean meal, and corn oil as PUFA sources supplemented with yeast and Curcuma xanthorrhiza

Roxb, stored for 2, 4, and 6 weeks

MATERIALS AND METHODS

Feed Supplements Preparation

Yeast supplement was prepared by modification on a procedure of Pusbangtepa (1981). The ingredients were 500g of rice flour, 50g of cassava tuber,10.5%of sugar, 10g of garlic, 20.5%of Alpinia galanga Sw, 10g of lemon juice, 10g of local (Bengkulu, Indonesia) yeast, and 500g of water; mixed, mounted in 10g each, then sun dried. This yeast contained 3.6 107 cfu/g. These are ingredients and steps in preparing the yeast; shown in Figure 2.1.

(a) (b) (c) (d)

Figure 2.1. Ingredients and steps in preparing the yeast: (a) yeast ingredients; (b) mixing; (c) dough; (d) mouting-drying

Curcuma supplement was made of C. xanthorrhiza Roxb tuber which was sliced thinly (about 1 mm thick), sun dried for about 4 hours, then oven dried in 60° C for 48 hours. It was grounded and refined as powder, which was then about 26.6% (w/w) out of fresh curcuma. Bio actives detected in this curcuma were 0.8% curcumin and 1.58% tannin. In this treatment, the curcuma powder was added as much as 2% or 20g/kg of PUFA- concentrate, mixed thoroughly. Steps in preparing the curcuma powder were presented in Figure 2.2.

PUFA- Concentrate Preparation

The concentratewas designed for lactating dairy goat with 30 kg of body weight and 1kg of milk production (NRC 1981). There were several ingredients combined in this concentrate as can be seen in Table 2.1.

Table 2.1 The ingredients of PUFA-concentrate supplemented with yeast and C. xanthorrhiza Roxb

Ingredients (% DM) PC0 PCY PCC PCM

Rice bran 35 35 35 35

Ground corn1 30 30 30 30

Cassava meal 15 15 15 15

Soy bean meal2 15 15 15 15

Corn oil 4 4 4 4

Mineral mix 1 1 1 1

Yeast - 0.5 - 0.5

Curcuma - - 2 2

Nutrient Calculated Requirement3

Dry matter (%) 84.85 2.7

TDN (%) 69.10 70.40

DE (Mcal/kg) 3.21 3.10

NEL (Mcal/kg) 1.60 1.42

Crude protein (%) 14.37 13.90

Ether extract (%) 3.14 NA

Crude fiber (%) 5.54 NA

Ca (%) 0.42 0.40

P (%) 0.29 0.28

PC0: PUFA concentrate with no supplementation; PCY: PUFA concentrate with 0.5%yeast; PCC: PUFA

-concentrate with no yeast and 2%curcuma; PCM:PUFA --concentrate with 0.5%yeast and 2%curcuma; 1half

roasted; 2 all roasted; 3 for 30kg dairy goat with 1kg of milk production (NRC 1981). NA: not available.

The ground corn was half roasted, while the soy bean meal was all. Both were roasted in 80° C for about 25 minutes until it was becoming light brown. Together with corn oil, these roasted soy bean meal, and roasted ground corn were intended as PUFA sources in concentrate, containing 46.46%, 50.35%, and 34.69% total fatty acid, respectively. Fatty acid contents of corn oil, roasted soybean meal, and roasted ground corn are presented in Table 2.2.

Cassava meal was prepared from the fresh tubers, sliced about 1 mm- thick, sun dried, then grinded as powder. Mixing of the ingredients were started by the smallest portion, manually as homogenized as possible. Each supplement was then added accordingly for each treatment.

Treatments and Experimental Design

There were four treatments as PC0: PUFA -concentrate with no supplementation; PCY: PUFA concentrate with 0.5% yeast; PCC: PUFA -concentrate with no yeast and 2%curcuma; PCM: PUFA - -concentrate with 0.5% yeast and 2% curcuma. These four PUFA - concentrates were then stored for 2, 4, and 6 weeks with three replications each. All together were 12 experimental units. The application of treatments utilized in this research was in Completely

Randomized Design with repeated measurement and split plot statistical analysis. Data were tabulated and analyzed for variance with any differences were detected by Duncan Multiple Range Test in significances of P<0.05 or P<0.01 according to Lentner and Bishop (1986).

Table 2.2 Fatty acid contents of corn oil, roasted soybean meal, and roasted ground corn

Fatty acid (% of total fat) Corn oil Roasted SBM Roasted ground corn

Lauric Acid, C12:0 nd nd 0.13

Myristic Acid, C14:0 0.03 0.06 0.14

Palmitic Acid, C16:0 8.98 8.32 9.9

Palmitoleic Acid, C16:1 0.09 0.07 0.11

Heptadecanoic Acid, C17:0 0.06 0.08 0.07

Stearic Acid, C18:0 1.64 3.16 2.59

Oleic Acid, C18:1n9c 23.81 15.36 21.95

Linoleic Acid, C18:2n6c 45.58 44.21 34.69

Arachidic Acid, C20:0 0.36 0.24 0.63

Cis-11-Eicosenoic Acid, C20:1 0.22 0.11 0.23

Linolenic Acid, C18:3n3 0.81 6.09 nd

Cis-11,14-Eicosedienoic Acid, C20:2 0.07 0.05 nd

Heneicosanoic Acid, C21:0 nd nd 0.03

Behenic Acid, C22:0 0.12 0.25 0.32

Erucic Acid, C22:1n9 0.02 nd nd

Tricosanoic Acid, C23:0 nd 0.03 nd

Lignoceric Acid, C24:0 0.14 0.08 0.3

Total Fatty Acid 81.93 78.12 71.85

Total SCFA (C4- C10) nd nd nd

Total MCFA (C12- C16) 9.1 8.45 10.28

Total LCFA (C>C16) 72.83 69.66 60.81

Total MUFA 23.92 15.43 22.06

Total PUFA(P) 46.46 50.35 34.69

Total Saturated fat (S) 11.33 12.22 14.11

Total Unsaturated (U) 88.67 87.78 85.89

Ratio P/S 4.10 4.12 2.46

Ratio U/S 7.83 7.18 6.09

n6/n3 56.27 7.26 nd

Atherogenicity 0.41 0.39 0.60

Analysis of Feed

Nutrient analyses of dry matter, crude protein (CP), crude fiber (CF), and ether extract (EE) were determined according to AOAC (2000). While, NFE was calculated as 100% - (moisture + ash + EE + CP + CF). The content of ADF (Acid Detergent Fiber) were analyzed by the method of Goering and Van Soest (1970). Minerals of Ca and P were analyzed using Atomic Absorbance Spectrophotometer (AAS) according to the procedure of AOAC (2000).

Bioactive analyses of curcumin was determined by maceration of 500g C. xanthorrhiza Roxb in 96% ethanol for 48 hours. The result of this maceration

was then extracted by soxhlet method in 96% ethanol, continued with evaporation using rotavapor. Residue from this process was washed with aquadest then centrifuged; the solid was crystallized by adding MeOH. This crystal with orange- brown color is curcuminoid, in which curcumin was part of it (Sutrisno et al. 2008). Tannin was analyzed by extracting 3g of sample in boiling water, then strained and added with 1% FeCl3. Whenever green, blue, and black colors were

detected in Spectrophotometer , this means positive indicator for tannin.

Microbial analyses for bacteria population was quantified by weighing 1g of fineground sample, diluted in 10 ml of aquadest. This solution was serially diluted in 6 folds (106), then 0.1 ml was taken appropriately and inoculated onto 10 ml NA (Nutrient Agar) plate, made in double. Plates were incubated unaerobic at 400C for 2 days. Fungi analysis was conducted with the same procedure, except it was inoculated onto 10 ml PDA (Potato Dextrose Agar). Population of colony forming unit (cfu) found in each plate divided by sample weight timed dilution frequencies. The mean value from the double plate counts was used for statistical analysis.

Fatty acid methyl esters (FAME) were determined by using gas chromatography (GC) Shimadzu 2010 series. There are several calculations of total short chain fatty acid (C4- C10), medium chain fatty acid (C12-C16), long chain fatty acid (C>16), mono unsaturated, poly unsaturated, saturated, unsaturated, ratio of PUFA/saturated, and unsaturated/saturated fatty acid. Fatty acid ratio of n-6/n-3 was calculated using this formula (Schmidely et al. 2005 ): n6/n3 = (linoleic acid + arachidonic acid)/linolenic acid

While, atherogenicity score was calculated with this formula (Ulbrict and Southgate 1991):

Atherogenicity index : (C12 + 4C14 + C16)

(total unsaturated fat)

These all analyses were conducted in week -2 and week- 6 of the storage of PUFA- concentrate. Temperatures were recorded three times (at 06 am, 12 noon, and 03 pm) daily for 6 weeks, using thermometer. Averages of temperatures during the 2 and 6 weeks were averaged.

RESULTS AND DISCUSSION

Moisture, Dry and Organic Matter

PUFA- concentrate supplemented with yeast and C. xanthorrhiza Roxb stored for 2- 6 weeks are presented in several tables. In Table 2.3 most of the treatments were higher in moisture (16.38%), lower in dry matter (-2.05%), and lower in organic matter (-2.81%) significantly (P<0.05) during 6 weeks of storage, compared to those in week 2 and week 4 in three treatments, except PUFA- concentrate supplemented with yeast and curcuma (PCM).

Table 2.3 Moisture, dry matter, organic matter, and ash in PUFA-concentrate supplemented with yeast and C. xanthorrhiza Roxb stored for 2-6 weeks

Nutrient PC0 PCY PCC PCM Average

Moisture (%)

2 weeks 8.46x 8.78x 8.79x 8.79 8.70X 4 weeks 8.24x 8.48x 8.41x 8.16 8.32X 6 weeks 11.16y 10.77y 11.41y 8.78 10.53Y

Average 9.29 9.35 9.53 8.58

Dry matter (%)

2 weeks 91.54x 91.22x 91.22x 91.21 91.30XY 4 weeks 91.77x 91.52x 91.59x 91.84 91.68X 6 weeks 88.84y 89.23y 88.60y 91.22 89.47Y Average 90.71 90.65 90.47 91.42

Ash (%)

2 weeks 6.88 6.66 6.35 6.63 6.63

4 weeks 6.57 7.31 6.94 7.05 6.97

6 weeks 6.95 6.78 8.07 6.84 7.16

Average 6.80 6.91 7.12 6.84

Organic matter (%)

2 weeks 84.66x 84.56x 84.87x 84.58 84.67X 4 weeks 85.20x 84.21x 84.65x 84.79 84.71X 6 weeks 81.89yab 82.45yb 80.52ya 84.38c 82.31Y Average 83.91AB 83.74A 83.35A 84.58B

These relationships showed that the longer the concentrates stored, the higher the water produced from microbial activity in organic materials; therefore the moisture was increasing and at the same time the dry and organic matter were decreasing. However, the PUFA- concentrate supplemented with yeast and curcuma (PCM) seemed to remain stable in moisture, dry matter, ash, and organic matter composition during the storage of 2, 4, and 6 weeks. In spite of these facts, this treatment was showing significantly (p<0.05) lowest moisture (8.78%), highest dry matter (91.22%), and the highest organic matter (84.38%) in 6 weeks of storage compared to those in week 2 of storage among treatments. Based on these data, the PUFA- concentrate supplemented with yeast and C. xanthorrhiza

Roxb (PCM) with the highest organic matter is considered the most sustained along with this period of storage (6 weeks). This meant that supplementation of yeast and curcuma powder stabilized moisture, dry and organic matters of the PUFA- concentrate.

Moisture content of the concentrate would be another important reason for the reduction in colony forming unit later. This is in conjunction with data on the viability of the cells as it is displayed in Table 2.7; that it showed a positive relationship when the lowest moisture happened in the 6 weeks of storage resulted in the highest concentration of Saccharomyces cereviseae as shown in PUFA- concentrate supplemented with yeast and curcuma (PCM). Increasing water availability and high temperature will activate yeast cells that eventually will be ceased because of inadequate available nutrients (Sullivan and Bradford 2011). Results in this experiment approved that dry matter and organic matter were decreasing with higher moisture content in the PUFA- concentrate with no supplements, yeast, or curcuma only, resulting in lower concentrations of S. cereviseae.

Ether Extract, Crude Protein, and Gross Energy

Other nutrients, such as crude protein, N-free extract (NFE), and gross energy in PUFA- concentrate supplemented with yeast and curcuma were not significantly changed within 2- 6 weeks of storage (Table 2.4).

Table 2.4 Ether extract, crude protein, crude fiber, N-free extract, and gross energy in PUFA-concentrate supplemented with yeast and C. xanthorrhiza Roxbstored for 2-6 weeks

Nutrient PC0 PCY PCC PCM Average

Ether extract (%)

2 weeks 8.96 8.41 8.98 8.17 8.63

4 weeks 8.09 8.28 9.05 9.05 8.62

6 weeks 8.10 8.14 8.56 8.33 8.28

Average 8.38 8.28 8.86 8.51

Crude protein (%)

2 weeks 12.26 12.00 11.79 11.60 11.91 4 weeks 11.54 11.65 11.69 11.58 11.62 6 weeks 11.86 11.27 11.76 11.32 11.55 Average 11.89 11.64 11.74 11.50

Crude fiber (%)

2 weeks 13.51x 13.30 13.14 13.48 13.36X 4 weeks 12.91xy 12.83 13.06 12.88 12.92XY 6 weeks 12.57y 12.15 12.64 12.12 12.37Y Average 13.00 12.76 12.95 12.83

NFE (%)

2 weeks 49.94x 50.85 50.96x 51.33 50.77XY 4 weeks 52.66y 51.45 50.85x 51.28 51.56X 6 weeks 49.36xab 50.89bc 47.57ya 52.60c 50.11Y Average 50.65A 51.07AB 49.79A 51.74B

Gross energy (kcal/g)

2 weeks 4124 4091 3986 4378 4145

4 weeks 3992 4067 3946 4023 4007

6 weeks 3974 3825 3752 3915 3867

Average 4030 3994 3895 4105

Different superscript in a row (a,b,c) or within column (x,y,z), different significantly (P< 0.05). PC0: PUFA concentrate with no supplementation; PCY: PUFA concentrate with 0.5% yeast; PCC: PUFA -concentrate with no yeast and 2% curcuma; PCM:PUFA --concentrate with 0.5% yeast and 2% curcuma.

However, ether extract contents in week-2 of PC0 (8.96%), PCY (8.41%), and PCC (8.98%) were significantly higher (P<0.05) than those in week- 6. While, in PCM concentrate, its ether extract was remained relatively the same during 2- 6 weeks storage. However, this concentrate showed the lowest ether extract in the 2 weeks storage. The crude fiber and NFE of 2 and 4 weeks storage were higher significantly (p<0.05) than that of in 6 weeks. Reductions were quantitatively found in crude protein (-8.01%) and gross energy (-6.65%) within 6 weeks of storage of all PUFA- concentrates. These data suggested that the longer the PUFA- concentrates (with or without yeast and curcuma supplementation) stored, the less nutritional values shown.

In conjunction with these facts (Table 2.3) that the average moisture content was increasing while the organic and dry matters were decreasing; consequently, other nutritional contents within the organic matter such as ether extract, crude fiber, crude protein, and gross energy were found diminishing as well. This suggested that during storage, fermentation process was happening where nutrients were broken down significantly. These carbon contents in the nutrients were being taken up for microbial growth, such as it happened in coincidence with higher population of Saccharomyces sp (2.35 107 cfu/g ) and Bacillus sp (11.43 107 cfu/g) as it can be seen in the next table .

Acid Detergent Fiber, Ca, and P

Within 2 weeks of storage, ADF contents of all PUFA- concentrates were not significantly different as can be seen in Table 2.5. Only ADF was affected, in week 2 and 4 were higher (P<0.05) than that of during 6 weeks of storage.

Table 2.5 Neutral detergent fiber, ADF, Ca, and P in PUFA-concentrate supplemented with yeast and C. xanthorrhiza Roxb stored for

2-6 weeks

Nutrient PC0 PCY PCC PCM Average

ADF (%)

2 weeks 25.16x 24.73 23.84 25.37 24.78X 4 weeks 36.38y 30.47 27.06 29.13 30.76Y 6 weeks 33.95xy 25.16 24.06 22.54 26.43XY Average 31.83B 26.78A 24.99A 25.68A

Ca (%)

2 weeks 0.38 0.36 0.44 0.47 0.41

4 weeks 0.59 0.57 0.70 0.74 0.65

6 weeks 0.52 0.53 0.57 0.60 0.56

Average 0.50 0.49 0.57 0.60

P (%)

2 weeks 0.66 0.62 0.61 0.63 0.63

4 weeks 0.45 0.55 0.60 0.61 0.55

6 weeks 0.64 0.67 0.64 0.75 0.68

Average 0.58 0.61 0.62 0.66

Different superscript in a row (a,b,c) or within column (x,y,z) , different significantly (P< 0.05). PC0: PUFA -concentrate with no supplementation; PCY: PUFA -concentrate with 0.5% yeast; PCC: PUFA concentrate with no yeast and 2% curcuma; PCM:PUFA -concentrate with 0.5% yeast and 2% curcuma.

In this week, PUFA- concentrate without any supplement (PC0) showed the highest ADF (33.95%). While, the lowest ADF (22.54%) carried by PUFA- concentrate with both supplements (PCM). In all treatments, the differences between week-2 and week-6 of storages were found in ADF (4.35%), Ca (32.84), and P (7.23%).

In overall, all three treated PUFA- concentrates were lower (P<0.05) in ADF compared to that of in control diet. Suggesting that these supplements (yeast, curcuma, and their combination) worked well in reducing fiber. The PCM was considered the most nutritional since it showed significantly (P<0.05) the lowest ADF with the highest Ca and P. Other study showed that basal diet for dairy cows in which 56g XP yeast incorporated, was having 31% NDF, 20.4% ADF, 0.96% Ca, and 0.39% P (Hristov et al. 2010). Compared to our data for, the NDF and P were more than doubled, while Ca was one third of those in XP diet.

Tannin and Curcumin

Bio actives detected in the C. xanthorrhiza Roxb used in this experiment were 0.8% curcumin and 1.58% tannin. Tannin levels in four treatments of PUFA- concentrate quantitatively were decreasing as much as 32.78% in 6 weeks storage compared to those in 2 weeks storage (Table 2. 6). Among the treatments, PUFA- concentrate with no supplements (PC0) quantitatively contained the lowest tannin; while PUFA- concentrate with yeast and curcuma supplements (PCM) had the highest one during 2- 6 weeks of storage. This suggested that curcuma powder contributed some amount of tannin in the concentrate. Tannins in all type of concentrate found in this study were not as high as it was in polyherbal combination (3.69%) as reported by Mirzaei and Prasad (2011).

Curcumin levels in both treatments, PCC and PCM were found the same in week -2, yet decreasing in week-6; however, it was slightly higher in PCM. These results showed that tannin and curcumin concentrations were found not as much high as than those in their original sources when other feedstuffs were combined together and depended upon its level added as well. Curcuminoids in

Table 2. 6 Tannin and curcumin in PUFA-concentrate containing yeast and

C. xanthorrhiza, Roxb during 2-6 weeks storage

Bioactive PC0 PCY PCC PCM Average

Tannin (%)

2 weeks 0.23 0.27 0.29 0.30 0.27

4 weeks 0.20 0.20 0.19 0.19 0.20

6 weeks 0.17 0.18 0.24 0.24 0.21

Average 0.20 0.22 0.24 0.24

Curcumin (%)

2 weeks - - 0.10 0.10 0.10

4 weeks 0.06 0.06 0.06

6 weeks - - 0.07 0.09 0.08

Average 0.08 0.08

PC0: PUFA -concentrate with no supplementation; PCY: PUFA -concentrate with 0.5% yeast; PCC: PUFA -concentrate with no yeast and 2% curcuma; PCM:PUFA -concentrate with 0.5% yeast and 2%

C. longa contained curcumin (1.06- 5.65%), demethoxycurcumin (0.83- 3.36%), and bisdemethoxycurcumin (0.42 – 2.16%) as observed by Jayaprakasha et al. (2002).

Microbial Viability

This self made yeast contained 3.6 107 cfu/g. The viable cell was within the range of other commercial yeast products (2.93 107 – 1.43 1013 cfu/g) that had been surveyed by Sullivan and Bradford (2011). They reported that there was a decrease in the viability of the cell approximately 90% per month when the active dry yeast was kept under high temperature of 400C; however, there was no significant difference of viable cells stored during spring and summer as the variability of the samples were so wide due to different distribution.

In previous research, the yeast was prepared in the ambient temperature of around 310C then added into PUFA- concentrate and kept in another ambient temperature of around 26.940C with average moisture of 78.83. These places of yeast preparation were certainly lower in the ambient temperature compared to the previous temperature.

Results in Table 2.7 showed that Saccharomyces sp was not significantly different within 2 weeks and 6 weeks; however, they were significantly (P<0.05) higher in 6 weeks compared to those in 2 weeks of storage.

Being the highest one was PUFA- concentrate with yeast and curcuma or PCM (23.5 106 cfu/g) or 2.35 107 cfu/g in 6 weeks of storage. This amount of viable cell seemed to be well protected by the addition of curcumin in the concentrate, while it was stored under suitable ambient temperature. This kind of concentrate would be beneficial for ruminants.

Bacillus sp were found significantly different (P<0.05) within 2 and 4 toward 6 weeks, while population in week 4 was about the same as in week 2. However, diet with combination of yeast and curcuma (PCM) in 4 weeks showed the highest (P<0.05) Bacillus sp. This might happen as there was yeast that could

Table 2.7 Microbes population in PUFA-concentrate supplemented with yeast and C. xanthorrhiza Roxb stored for 2-6 weeks

Microbe PC0 PCY PCC PCM Average

(106 cfu/g)

Saccharomyces sp

2 weeks 1.57x 1.20x 2.43x 4.00x 2.30X 4 weeks 0.07x 1.03x 0.43x 5.87x 1.85X 6 weeks 18.83y 19.77y 20.63y 23.50y 20.68Y

Average 6.82 7.33 7.83 11.12

Bacillus sp

2 weeks 6.93 3.67 0.37 9.20 5.04

4 weeks 5.90a 2.90a 1.93a 28.73b 9.87 6 weeks 19.47 8.43 7.87 11.43 11.80 Average 10.77AB 5.00A 3.39A 16.46B

Different superscript in a row (a,b,c) or within column (x, y, z), different significantly (P < 0.05). PC0: PUFA concentrate with no supplementation; PCY: PUFA concentrate with 0.5% yeast; PCC: PUFA -concentrate with no yeast and 2% curcuma; PCM: PUFA --concentrate with 0.5% yeast and 2% curcuma.

62

Sulistyowati E, Santoso U, Badarina I. 2011. Supplementation of concentrate with different levels of temulawak (Curcuma xanthorriza Roxb) on milk production of lactating Frisien Holland cows. Proceedings: The 2nd International Symposium on Temulawak- the 40th Meeting on National Working Group on Indonesian Medicinal Plant. ISBN: 978-979-25-1209-0. Pp: 116- 120.

Sullivan ML, Bradford BJ. Viable cell yield from active dry yeast products and effect of storage temperature and diluents on yeast viability. J Dairy Sci. 94:526-531.

Suriyasathaporn W. 2010. Milk quality antimicrobial resistance against mastitis pathogens after changing from a conventional to an experimentally organic dairy farm. Asian- Aust J Anim Sci. 23 (5): 659- 664.

Sutrisno, Sukarianingsih D, Saiful M, Putrika A, Kusumaningtyas DI. 2008. Curcuminoids from Curcuma xanthorrhiza Roxb: isolation, characterization, identification. Proceedings of the First International Symposium on Temulawak. IICC. Bogor (ID). May 27- 29. ISBN No 978-979-25-1207-6. Pp: 225- 230.

Teles FFF, Young CK, Stull JW. 1978. A Method for rapid determination of lactose. J Dairy Sci. 61: 506- 508.

Thiruvenkadan AK. 2012. Improvement in rural livelihood through dairy goat farming in India. Proceeding of the 1st Asia Dairy Goat Conference, Kuala Lumpur, Malaysia 9-12 2012. 174- 178.

Tilley JMA, Terry RA. 1963. A two stage technique for the in vivo digestion of forage crops. J. of the British Grassland Society. 18: 104-111.

Ulbricht TLV and Southgat DAT. 1991. Coronary heart disease: Seven dietary factors. Lancet. 338:985-992,

Van Soest PJ, Robertson JB, and Lewis BA. 1991. Methods for dietary fiber, neutral detergent fiber and non starch polysaccharides in relation to animal nutrition. J Dairy Sci. 74:3583- 3592.

Wallace RJ. 1994. Ruminal microbiology, biotechnology, and ruminal nutrition: progress and problems. J Anim Sci. 72:2992-3003.

Weinstein M. 2005. Contribution of Wild and Domestic Large Mammals over Time on the great Hungarian Plain: “introduced” sheep and goat, local domesticated cattle and pigs, and large wild animals (aurochs, wild pigs, red deer) by cultural period. Ohio State Univ. Periodicals. http://www.jyi.org/research.

Whitlock LA, Schingoethe DJ, Hippen AR, Kalscheur KF, Baer RJ, Ramasmy N, Kasperson KM. 2002. Fish oil and extruded soybeans fed in combination increase conjugated linoleic acids in milk of dairy cows more than fed separately, J Dairy Sci., 85:234-243.

Wientarsih I and Meulen U. 2008. The effect of diets varying in curcuma (C. Curcuma xanthorrhiza, Roxb) on blood plasma LDL- peroxidation in rabbits. Proceedings of the First International Symposium on Temulawak. IICC. Bogor (ID). May 27- 29. ISBN No 978-979-25-1207-6. Pp: 313-316.

William PEV, Tait CAG, Innes GM and Newbold CJ. 1991. Effects of the inclusion of yeast culture (Saccharomyces cerevisiae plus growth medium) in the diet of dairy cows on milk yield and forage degradation

63

and fermentation patterns in the rumen of steers. J Anim Sci. 69:3016- 3026.

Yalçın S, Pınar C, Arif O, Gürdal, Bağc C, and Eltan O. 2011. The nutritive value of live yeast culture (Saccharomyces cerevisiae) and its effect on milk yield, milk composition and some blood parameters of dairy cows. Asian- Aust J Anim Sci 24 (10): 1377-1385.

Zhong R, Xiao W, Ren G, Zhou D, Tan C, Tan Z, Han X, Tang S, Zhou C, and Wang M. 2011. Dietary tea catechin inclusion changes plasma biochemical parameters, hormone concentrations and glutathione redox status in goats. Asian-Aust. J Anim Sci. 24 (12):1681-1689.

RINGKASAN

ENDANG SULISTYOWATI. Fermentasi Rumen, Produksi dan Kualitas Asam Lemak Susu pada Kambing Perah dengan Fase Laktasi Akhir yang

Diberi Diet –PUFA yang Disuplementasi Ragi dan C. xanthorrhiza Roxb

Dibimbing oleh TOTO TOHARMAT, ASEP SUDARMAN dan KOMANG G. WIRYAWAN.

RINGKASAN

Dalam rangka memenuhi kebutuhan susu nasional dan pada saat yang sama memperbaiki kualitas susu, dalam hal ini asam lemak dari ternak ruminan laktasi seperti kambing perah, diperlukan perbaikan nutrisional dalam ransum. Hasil- hasil penelitian sebelumnya telah dilakukan pada kambing atau sapi perah yang diberi suplemen ragi, pasta tape- temulawak atau curcuma (C. xanthorrhiza Roxb), Tabut blok, dan konsentrat laktasi yang mengandung ragi dan temulawak dapat meningkatkan produksi susu dan memperbaiki kualitas susu. Oleh karena itu, tiga penelitian yang meliputi evaluasi nutrisi konsentrat-PUFA (polyunsaturated fatty acid) selama penyimpanan, analisis ransum-PUFA secara fermentasi in vitro, dan aplikasi ransum- PUFA secara in vivo telah dilakukan. Ransum yang mengandung sumber PUFA, yaitu jagung giling sangrai, tepung kedelai sangrai, dan minyak jagung yang disuplementasi ragi Saccharomyces cereviseae dan curcuma diaplikasikan pada kambing perah dengan fase laktasi akhir.

Penelitian pertama bertujuan untuk mengevaluasi konsentrat- PUFA dengan suplementasi ragi dan curcuma yang disimpan selama 2- 6 minggu. Perlakuan konsentrat- PUFA yaitu tanpa suplemen ragi dan curcuma (PC0), 0.5% ragi (PCY), 2% curcuma (PCC), dan 0.5% ragi dan 2% curcuma (PCM). Ragi (mengandung 3.6 x 107 cfu/g) dan curcuma (mengandung 0.8% curcumin dan 1.58% tannin) disiapkan sendiri. Hasil menunjukkan bahwa kandungan kadar air dan S. cereviseae (20.68 x 106 cfu/g) meningkat nyata (P<0.05) pada penyimpanan 6 minggu; sedangkan, bahan kering (BK), bahan organik (BO), serat kasar (SK), dan bahan ekstrak tanpa nitrogen (Beta-N) menurun nyata (P<0.05). Total PUFA (P), P/S, mono unsaturated fatty acid (MUFA), dan long chain fatty acid (LCFA) lebih tinggi pada konsentrat- PUFA dengan 2% curcuma (PCC). Sebaliknya, pada konsentrat- PUFA dengan 0.5% ragi dan 2% curcuma (PCM) tinggi lemak tak jenuh dan rasio U/S. Secara keseluruhan, digabung dengan level nutrisi lainnya, pada penyimpanan 2- 6 minggu, konsentrat- PUFA dengan kombinasi 0.5% ragi dan 2% curcuma (PCM) adalah yang paling bisa dipertimbangkan secara nutrisi dan kesehatan.

Penelitian kedua, in vitro, dilakukan untuk mengevaluasi performa fermentasi rumen dari ransum-PUFA (mengandung konsentrat-PUFA, limbah kedelai dari industry tahu dan rumput Gajah) yang disuplementasi ragi dan C. xanthorrhiza Roxb. Rancangan percobaan yang diaplikasikan adalah Rancangan Kelompok dengan 4 perlakuan dengan 4 cairan rumen dari empat ekor kambing yang dipotong dan 3 ulangan masing-masing. Perlakuan yang diberikan adalah tanpa suplemen (PD0); 0.5% ragi (PDY); 2% curcuma (PDC); dan kombinasi 0.5% ragi dan 2% curcuma (PDM ). Variabel yang dianalisis adalah pH,

kecernaan bahan kering (KCBK) dan kecernaan bahan organik (KCBO), N-NH3, VFA total dan parsial, TPC, dan CH4. Hasil menunjukkan bahwa, walaupun KCBK- KCBO dan N- NH3 terendah; namun, didukung dengan populasi protozoa yang rendah dan produksi VFA yang tinggi, ransum-PUFA dengan ragi dan curcuma bisa dipertimbangkan sebagai ransum yang paling potential dalam meningkatkan metabolism nutrien dalam rumen.

Penelitian ketiga, in vivo, dilakukan untuk mengevaluasi performan ransum-PUFA dengan suplementasi ragi dan curcuma terhadap produksi dan komposisi nutrisi serta asam lemak susu, mastitis status, kecernaan nutrien, dan metabolit darah pada 20 ekor Peranakan Ettawa (PE) pada fase laktasi akhir (sekitar 4 bulan laktasi). Rancangan perlakuan berdasarkan pengelompokan tingkat produksi susu dengan menggunakan Rancangan Kelompok dengan 5 perlakuan dan 4 ulangan. Perlakuan terdiri atas tanpa aditif (PD0), 3 tablet laktasi Asifit (PDA), 0.5% yeast (PDY), 2% curcuma (PDC), dan kombinasi 0.5% yeast dan 2% curcuma (PDM) yang ditambahkan sebagai topping menjelang pemberian makan pagi (06.00) dan sore (15.00); Asifit diberikan per oral pagi hari. Sumber PUFA selain konsentrat- PUFA dari penelitian sebelumnya, juga dicampurkan kulit kedele dari limbah produksi tahu. Produksi susu dicatat sebelum, selama, dan setelah perlakuan. Perlakuan PDC mengandung lemak lebih rendah dibanding PDM, dan lebih tinggi serat kasar tetapi rendah ADF dan Phosphor (P). Pada ransum- PUFA dengan ragi (PDY) menunjukkan rendah protein. Produksi susu selama perlakuan secara kuantitatif tinggi pada PDA dan PDY. Pada pasca perlakuan, produksi ini lebih tinggi (P<0.05) dibanding perlakuan PDY dan PDC; sedangkan PDM berada diantaranya. Komposisi susu pada PDM secara kuantitas lebih rendah untuk lemak, 4% FCM, % dan berat protein, rasio lemak: protein, bahan kering, dan bahan kering tanpa lemak (BKTL), namun lebih tinggi untuk laktosa dan Ca. Indikator mastitis dan metabolit darah tidak terpengaruh (P>0.05) oleh perlakuan. Namun, secara kuantitas PDM menujukkan angka terendah untuk SCC, terendah Hb, PCV,dan glukosa, prolaktin dan trigliserida. Pada perlakuan PDM, konsumsi nutrien lemak antarperlakuan tertinggi (P<0.05), sementara, kecernaan nutrisi sebagian besar juga dipengaruhi oleh perlakuan (P<0.05 atau P<0.01). Perlakuan ransum-PUFA dengan curcuma (PDC) menunjukkan paling rendah untuk BK, BO, PK, LK, dan GE (gross energy); sementara, PDY dan PDM tinggi dalam kecernaan nutrisi. Disimpulkan bahwa 0.5% ragi dan 2% curcuma bubuk yang ditambahkan pada ransum-PUFA (PDM) adalah yang terbaik dalam hal nutrisi dan metabolit serta produksi susu pasca perlakuan (dengan semakin berlanjutnya fase laktasi) pada kambing perah.

Hasil penelitian yang berkaitan dengan asam lemak menunjukkan bahwa PDM mengandung tinggi asam lemak total, asam lemak rantai sedang, rantai panjang, dan asam lemak tak jenuh; namun, rendah dalam SCFA, n6/n3 rasio, dan indeks atherogenik. Kualitas susu kambing perah yang diberi pakan PUFA- diet dengan ragi dan curcuma ini dapat dipertimbangkan sebagai produk yang lebih sehat. Oleh karena itu, ransum PUFA dengan 0.5% ragi dan 2% curcuma merupakan ransum yang dipilih untuk diaplikasikan pada ternak kambing perah.

Kata kunci: C. xanthorrhiza Roxb, in vitro, in vivo, kambing perah,

ENDANG SULISTYOWATI. Ruminal Fermentation, Production and Fatty Acid Quality of Milk of Late Lactation Dairy Goat Fed PUFA-Diet

Supplemented with Yeast and C. xanthorrhiza _{Roxb. Supervised by TOTO}

TOHARMAT, ASEP SUDARMAN and KOMANG G. WIRYAWAN.

SUMMARY

In order to meet milk demand nationally and at the same time to improve milk quality, in this case fatty acid of lactating ruminants such as dairy goat; it is required a nutritional improvement in the ration. Based on previous research results that have been applied to either dairy goat or dairy cows with supplementations of yeast, fermented cassava- curcuma (C. xanthorrhiza roxb), Tabut block, and lactating concentrate were reported to increase milk yield and milk quality. Therefore, three experiments, evaluating nutrient contents during storage of the polyunsaturated fatty acid or PUFA- concentrate, analyzing fermented PUFA- diet in vitro, and applying PUFA- diet in vivo had been conducted. Diet containing PUFA- concentrate (PUFA sources consisted of roasted ground corn, roasted soybean meal, and corn oil) supplemented with yeast and curcuma had been applied in late lactation dairy goats.

The first experiment was to evaluate the nutritive values of PUFA- concentrate supplemented with yeast and curcuma stored in 2-6 weeks. There were four PUFA- concentrates, namely, no supplement (PC0), 0.5% yeast (PCY), 2% curcuma powder (PCC), and a mixture of 0.5% yeast with 2% curcuma (PCM). Yeast (containing 3.6 x 107 cfu/g) and curcuma powder (containing 0.8% curcumin) were self made. These concentrates were evaluated for nutrition and fatty acid contents during 2 and 6 weeks of storage. Results showed that moisture and Saccharomyces cereviseae (20.68 x 106 cfu/g) increased significantly (P<0.05) in 6 weeks of storage; whereas, dry matter (DM) and organic matter (OM), crude fiber (CF), nitrogen free extract (NFE) decreased significantly (P<0.05). The total PUFA (P), P/S, monounsaturated fatty acid (MUFA), and long chain fatty acid (LCFA) contents were found higher in PUFA- concentrate with 2% curcuma powder. Whereas, the PUFA- concentrate with 0.5% yeast and 2% curcuma powder was higher in unsaturated (U) fat and the ratio of U/S. Based on all nutrient values during the storage of 2- 6 weeks, the PUFA- concentrate with 0.5% yeast and 2% curcuma powder was considered nutritionally reasonable.

The second experiment, in vitro, was conducted to evaluate the ruminal performances of PUFA-diet (containing PUFA-concentrate with soybean byproduct and King grass) supplemented with yeast and curcuma. Experimental design applied was Randomized Complete Block of 4 ruminal liquor derived from four slaughtered goats and 4 treatments (PD0, PDY, PDC, PDM ), in 3 replications. Variables analyzed were pH, N-NH3, total and partial VFA, TPC, and CH4. Results showed that in spite of having the lowest organic and dry matter digestibilities as well as N- NH3; however, supported by low protozoa

population and high VFA production in the goat rumen fluid, the PUFA- diet with a mixture of yeast and curcuma additives was considered the most potential diet to improve nutrient metabolism in rumen.

The third experiment, in vivo, was to evaluate milk production and health of dairy goat fed PUFA-diet supplemented with yeast of curcuma. The were 20 crossbred Ettawa goats in the late lactation selected based on their production levels and grouped them in Randomized Block design to receive 5 treatments of no supplement (PD0), 3 tablets of Asifit (PDA), 0.5%/d yeast (PDY), 2%/d curcuma (PDC), and a mix of 0.5%/d yeast and 2%/d curcuma (PDM) in diets containing concentrate with PUFA sources (roasted ground corn, roasted soy bean meal, and corn oil), soybean by-product, and King Grass. The variables evaluated were milk yield, milk composition, mastitis status, nutrient digestibility, and blood metabolites. Milk yield was recorded daily pre, during, and post treatment. The PDC had lower ether extract than that of mix diet and showed higher crude fiber but lower ADF; diet with yeast had lower crude protein; and mix diet showed lower P content. Milk yield during treatment was higher in PDA and PDY. Post treatment, these milk yields were higher (P<0.05) than those in PDY and PDC; while the PDM was in between. There were tendencies that mix diet (PDM) had lower milk fat, 4% FCM, % protein and weight, fat: protein ratio, dry matter, and solid non fat, but it showed higher lactose and Ca percentage. Mastitis indicators and blood metabolites were not affected by treatments. However, PDM showed lowest SCC, higher Hb, PCV, and glucose but had lower prolactin and triglyceride. In PDM, nutrient intake of ether extract was the highest (P<0.05), while nutrient digestibilities were mostly affected significantly (P<0.05 or P<0.05). The PDC had the lowest DM, OM, CP, EE, and GE; whereas PDY and PDM showed higher digestibilities. As conclusion, a mix supplement of 0.5%/d dried yeast and 2%/d curcuma powder was considered reasonable since it showed a better recovery in milk yield after treatment with progressing lactation in dairy goat.

Results dealing with fatty acid demonstrated some consistencies in diet containing polyunsaturated fatty acid (PUFA) supplemented with 0.5% yeast and 2% curcuma (C. xanthorrhiza Roxb) that were high in total fatty acid, MCFA, LCFA, and PUFA. Milk fatty acid of goat fed with this diet showed high in LCFA and MUFA; while it was low in SCFA, n6/n3 ratio, and atherogenicity index. These milk qualities were optimally considered good in terms of healthier product. Therefore, the PUFA- diet with 0.5% yeast and 2% curcuma was a reasonable choice to be applied for dairy goat.

Key words: C. xanthorrhiza Roxb, dairy goat, in vitro, in vivo, PUFA- diet, Yeast, storage