IN VITRO GOAT FERMENTATION OF PUFA- DIET SUPPLEMENTED WITH YEAST AND C. xanthorrhiza Roxb

The AP ratio in these diets 3.54- 3.88 were about in the same range as that of with alfalfa hay 3.77- 3.81; whereas diet with high carbohydrate gave lower ratio of 1.87- 2.40. This has been known that type of diet will determine its production of propionate and acetate; carbohydrate produces higher propionate, while fiber will produce higher acetate. The AP ratio increased with the longer fermentation, suggesting that the more structural carbohydrate was degraded, thefore, produced higher acetate and lower propionate. Tannin contents in all PUFA-diets were relatively the same 0.487 - 0.491. Tannin and curcumin added in PDM yielded the highest total VFA 45.27 mM and the lowest isobutirate 0.66. A different result showed that tannin 13.84 from Pakar leaves Ficus infectoria fed to goats showed some lower variables, such as in total VFA 4mM and propionate 11.86; but higher in N-NH3 5.29mM, asetat 83.56, and total fungi 28.16 than those found in feed lower in tannin 8.6 as reported by Singh et al. 2011. Concentration of N- NH3 was decreasing from 11.26 mM in PD0 to 8.73 mM in PDM. This level of NH3 was much higher than that of NH3 8.5- 9.6 mM with pH of around 6.7, in goat fed with extruded soybean with or without NaHCO3 Schmidely et al., 2005. The N-NH3 concentration in dairy cow fed diet containing tannin extract was detected the lowest 5.1 mM compared to that of control diet 17.2 mM with pH around 7.05- 7.09 Khiaosa- Ard et al. 2008. Microbial Activity Total plate count TPC of the fermented diet Table 3.5 was affected decreasing by curcumin PDC or PDM compared to basal diet 107.28 10 5 cfuml. Protozoa population found numerically the lowest 7.90 10 3 cfuml was in PDM, while in control diet PD0 was the highest 32. 10 3 cfuml. These results suggested that yeast or curcuma or their combination worked well in protecting the PUFA- diet. Table 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 Variable PD0 PDY PDC PDM P pH 6.71 ± 0.02 6.73 ± 0.07 6.73 ± 0.05 6.74 ± 0.04 NS IVDMD 62.72 a ± 1.66 60.26 ab ± 1.90 60.77 a ± 1.63 58.00 b ± 1.05 IVOMD 64.85 a ± 2.22 61.74 bc ± 2.07 62.82 ab ± 1.36 59.63 c ± 0.59 NH3 mM 11.26 ± 3.82 10.28 ± 3.82 9.31 ± 3.75 8.73 ± 3.16 NS VFA total mM 42.13 ± 3.45 43.95 ± 11.09 44.82 ± 12.30 45.27 ±12.92 NS Acetate A, mM 67.13 ± 6.39 67.68 ± 4.43 68.82 ± 4.98 68.46 ± 5.73 NS Propionate P, mM 19.78 ± 4.24 19.35 ± 2.60 18.25 ± 3.20 18.64 ± 3.10 NS Butirate B, mM 11.33 ± 2.90 11.21 ± 3.23 11.13 ± 2.85 11.05 ± 3.86 NS Iso Butirate , mM 1.27 ± 0.17 1.11 ± 0.24 1.08 ± 0.18 0.66 ± 0.58 NS Valerate V, mM 1.76 ± 0.66 1.76 ± 0.81 1.80 ± 0.79 1.86 ± 0.85 NS IsoValerate , mM 2.66 ± 0.30 2.48 ± 0.58 2.46 ± 0.48 2.08 ± 0.87 NS A:P 3.54 ± 0.97 3.55 ± 0.55 3.88 ± 0.82 3.76 ± 0.77 NS PD0: PUFA- diet with no additives; PDY: PUFA- diet with 0.5 yeast; PDC: PUFA- diet with 2 curcuma; PDM: PUFA- diet with mix of 0.5 yeast and 2 curcuma. Averages with different superscript, differed significantly P0.05. NS: not significant Protozoa population in this research was lower than that found in diet added with saponin generated from Yucca schidigera 6.17 10 5 cfuml and Quillaja saponaria 6.17 10 5 cfuml in dairy cow ruminal fermentation Holshausen et al. 2009; while addition of tannin extracted from Acacia mearnsii was effectively very low 1.91 10 3 cfuml and the protozoa population could be higher when the additive was not in the form of extract Khiaosa-Ard et al. 2009. Supplementation of Hibiscus tiliaceus leaves saponin for 5 significantly decreased rumen protozoa population 4.50 10 3 cfuml, compared to control 16.25 10 3 cfuml and dose 20 6.75 10 3 cfuml as reported by Istiqomah et al. 2011. Total protozoa Entodinium spp, Isotrica spp., Dasytrica spp., Epidinium spp., Ophryoscolex spp., and Diplodinium spp. detected in rumen fermentation of dairy cow supplemented with yeast XP 56gd was much higher 8.03 10 5 cfuml as reported by Hristov et al. 2010. This described that type of plants or additives, the way to administer them, and the right dose will determine the effectiveness of the bioactives content during the ruminal fermentation. Gas production CH4 in a mix diet PDM and curcuma diet PDC tended to be higher around 13- 14, respectively, while the PDY and PD0 tended to produce lower about 12. This implied that supplementation of tannin and curcumin was not effective in reducing methan CH4 production in this study. On the other hand, Khiaosa- Ard et al. 2009 reported that extract tannin was able to decrease gas production methan, CO2, and H2 to the lowest 0.454 ld. Decreasing gas production was also found in diet containing silage and concentrate added with Peltiphyllum peltatum rich in phenol and tannin Jayanegara et al. 2010. Production of methan gas was reported higher 10.3 gd with XP yeast supplementation diet than that in control 9.7 gd in dairy cow Hristov et al. 2010. Methane gas was produced through a mechanism starting from anaerobic degradation; trigliceride hydrolysis, saturation of unsaturated fatty acid C18:2  C18:1 C18:0C16:0 C14:0, and successive β-oxidation of saturated fatty acid C10- C18. These all reactions produced acetate, then reacted with H2 producing CH4 Sage et al. 2008. Table 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 Variable PD0 PDY PDC PDM P TPC 10 5 cfuml 107.28 7.85 9.41 43.30 NS Protozoa10 3 cfuml 32.00 ± 24.70 28.50 ± 25.31 25.50± 24.95 7.90 ± 4.09 NS CH4 vv 12.90 ± 1.25 12.49 ± 1.33 14.20 ±2.79 13.43 ± 2.26 NS H2 vv 0.04 ± 0.02 0.06 ± 0.04 0.14 ± 0.17 0.04 ± 0.02 NS O2 vv 0.006 ± 0.00 0.006 ± 0.00 0.006 ± 0.00 0.006 ± 0.00 NS CO2 vv 87.07 ± 1.24 87.45 ± 1.30 85.67 ±2.82 86.51 ± 2.22 NS PD0: PUFA- diet with no additives; PDY: PUFA- diet with 0.5 yeast; PDC: PUFA- diet with 2 curcuma; PDM: PUFA- diet with mix of 0.5 yeast and 2 curcuma. NS: not significant CONCLUSION In spite of having the lowest organic and dry matter digestibilities as well as N- NH3; however, supported by having 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. 4. MILK YIELD AND COMPOSITION, AND BLOOD PARAMETERS OF LATE LACTATION DAIRY GOAT FED POLYUNSATURATED FATTY ACID DIET SUPPLEMENTED WITH YEAST AND C. xanthorrhiza Roxb ABSTRACT The third experiment, in vivo, was to evaluate milk production and dairy goat health fed PUFA-diet supplemented with yeast of curcuma. The application was using 20 crossbred Ettawa goats in the late lactation selected based on their production levels and grouped them in Randomized Block design to receive 5 dietary treatments. The treatments were no supplement PD0, 3 tablets of Asifit PDA, 5gd yeast PDY, 20gd curcuma PDC, and a mix of 5gd yeast and 20gd 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. Variables measured were milk yield and composition, mastitis status, nutrient digestibility, and blood metabolites. Milk yield was recorded daily pre, during, and post treatment. The PDC numerically 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 numerically was higher in PDA and PDY. Post treatment, these milk yields were higher P0.05 than those in PDY and PDC; while the PDM was in between. There were tendencies that numerically 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 resulted in lowest SCC, higher Hb, PCV, and glucose but had lower prolactin and triglyceride. In PDM, nutrient intake of ether extract was the highest P0.05, while nutrient digestibilities were mostly affected significantly P0.05 or P0.01. The PDC had the lowest DM, OM, CP, EE, and GE; whereas PDY and PDM showed higher digestibilities. As conclusion, a mix supplement of 5gd self made dried yeast and 20gd curcuma powder was considered reasonable since it showed a better recovery in milk yield after treatment with progressing lactation in dairy goat. Key words: curcuma, dairy goat health, milk yield and components, PUFA-diet, yeast. INTRODUCTION Goat is gaining its popularity as an alternative milk producer due to its nutritional aspect of its milk. Goat population in Indonesia reached 17,482,722 in 2011, with an increased of 20.82 of that in 2007 DGLAH 2011. Goat milk contributed as much as 35 toward world milk production and it has been an increase about 9 Weinstein 2005. Goat milk is higher in short chain fatty acid of C4- C10 and medium chain of C12- C16 Bouattour et al. 2008; whereas cow milk contains higher long chain fatty acid of C18 Bernal- Santos et al. 2010. Nutrient contents of feed determine milk composition and its product. In low forage and high concentrate diets, the low milk fat was improved and milk protein stabilized by the inclusion of 20 extruded soybean with the addition of sodium bicarbonate in midlactation goats Schmidely et al. 2005. Feeding of 4.6 soybean oil was reported to increase milk fat and milk fatty acid; however, dry matter intake, milk yield, body weight, and body condition score were not affected in dairy goats Bouattour et al. 2008. Yeast supplementation in ruminants has been reported with variable results, depending upon the nature of diets and yeast as well as the lactation stage. Wallace et al. 1994 described some increases in microbes population, fiber digestibility, protein microbes, feed intake, nutrient supply into digestive track, eventually improved production performance. Positive effects of yeast were found on rumen volatile fatty acid, dry matter intake, crude protein, and organic matter digestibility; however the impaired effect on decreasing lactic acid could be overcomed by increasing concentrate in diet Desnoyers et al. 2009. Improved nutrient digestibility and quantatively increased fat and energy milk were showed by yeast supplementation Saccharomyces cereviseae CNCM 1- 1077, 1.2 10 10 cfud in Holstein cows Bagheri et al. 2009, increased milk yield with 20 gd in Holstein cows Sulistyowati et al. 2010 b , and improved milk yield and milk fat with 50g RumiSacc in Holstein cows Yalcin et al. 2011. Medicinal herbs have been applied to improve health status and production performance as they have galactogouge property and digestive process as reported by Mirzaei and Prasad 2011 that low dose 125mgkg BW of polyherbal increased milk yield and kids performance. Curcuma xanthorrhiza Roxb powder 15gkg concentrate was optimal in improving milk yield and milk composition in Holstein cows Sulistyowati et al. 2011; whereas Sinaga et al. 2011 reported that curcumin extract 160 ppm of C. domestica improved energy digestibility of ration, decreased rate of passage such that the retention time was longer and increased absorption that finally improved body weight in pig. Curcumin was described to increase bile and lipase that will improve fat digestibility and decrease cholesterol; beside, as anti pathogen microbes, improve nutrient absorption then improve production performance Bauwman et al. 1983. Therefore, it would be beneficial to apply either yeast or curcuma or in combination in concentrate containing PUFA polyunsaturated fatty acid sources to evaluate milk yield, milk composition, nutrient digestibility, and blood metabolite in late lactation dairy goat. MATERIALS AND METHODS Yeast, Curcuma, and PUFA- Concentrate Analyses Yeast supplement was prepared by combining some ingredients, rice flour, cassava tuber, sugar, garlic, Alpinia galanga Sw, lemon juice, local yeast Bengkulu, Indonesia, and water based on modification of procedures of Pusbangtepa 1981. These ingredients were all mixed thoroughly, shaped in 10g each, let them dried under the sun for about 10 hours. This made dry yeast contained 3.6 10 7 cfug. Curcuma powder was made of C. xanthorrhiza Roxb tuber which was sliced thinly, sun dried for about 4 hours, then oven dried in 60° C for 48 hours, grounded and refined. The powder form was about 26.6 ww out of fresh curcuma. Curcumin and tannin in the powder form of C. xanthorrhiza Roxb were analyzed and contained 0.8 and 1.58, respectively. Diet that was used containing of the PUFA- concentrate from previous experiment combined with soybean by- product that was used to be fed to the dairy goat in the farm. The PUFA- concentrate was designated for 30 kg lactating dairy goat with 1kg of milk production NRC 1981. Ground corn was half roasted, while soy bean meal was all roasted in 80° C for about 25 minutes until turned light brown. These roasted soy bean meal, roasted ground corn, and corn oil were intended as PUFA sources containing of 50.35, 34.69, and 46.46 out of total FAME fatty acid methyl esters, respectively. Cassava meal was prepared from the fresh tubers, thin sliced, sun dried, then grinded as powder. Soybean by product was in the form of the skin of soybean after an extrusion from boiling it provided by small scale tofu industry. Mixing of the ingredients was started from the smallest portion, manually as homogenized as possible. Each treatment was prepared with an addition of respective supplement. Treatments, Experimental Design, and Animals Treatments were based on the basic diet containing the same PUFA- concentrate and soybean by- product as PUFA- diet PD. The PUFA- ration formula and nutrient contents are delivered in Table 4.1. Table 4.1 Ingredients and nutrient composition of PUFA-diet containing Asifit, yeast and C. xanthorrhiza Roxb for dairy goat Ingredients : PD0 PDA PDY PDC PDM KG Rice bran 18.42 18.42 18.42 18.42 18.42 - Ground corn 15.79 15.79 15.79 15.79 15.79 - Soy bean meal 7.89 7.89 7.89 7.89 7.89 - Cassava meal 7.89 7.89 7.89 7.89 7.89 - Soy bean by-product 27.49 27.49 27.49 27.49 27.49 - Corn oil 2.11 2.11 2.11 2.11 2.11 - Mineral 0.53 0.53 0.53 0.53 0.53 - King grass KG 19.88 19.88 19.88 19.88 19.88 - Asifit - 0.22 - - - - Yeast - - 0.5 - 0.5 Curcuma powder - - - 2.0 2.0 Nutrients DM basis: Drymatter 94.13 94.13 93.96 93.72 94.06 92.24 Ash 8.48 8.48 7.96 7.82 8.21 11.65 Crude protein 15.27 15.27 14.11 15.33 15.04 13.42 Ether extract 7.66 7.66 8.14 7.96 8.77 3.22 Crude fiber 13.30 13.30 14.57 18.60 14.51 31.32 NFE 49.12 49.12 49.18 44.0 47.60 32.64 ADF 29.07 29.07 29.54 48.18 29.35 78.99 Gross energyMcalkg 3.84 3.84 3.86 3.83 3.90 3.71 Ca 0.61 0.61 1.20 1.02 0.84 0.7 P 0.47 0.47 0.53 0.57 0.29 0.13 CaP 1.30 1.30 2.26 1.79 2.90 0.64 Tannin 0.487 0.487 0.492 0.491 0.491 - Curcumin - - - 0.13 0.11 - S. cereviseae10 -6 cfug 1.6 1.6 7.3 1.2 4.0 - PUFA- diet without supplements PD0, with 3 Asifit tablets PDA, 0.5 yeast PDY, 2 curcuma PDC, mix of 0.5 yeast and 2 curcuma PDM Basal diet was a mixture of PUFA- concentrate and soybean by- product as PUFA- diet PD. The basal diet without supplements designated as PD0, added with 3 tabletsdairy goat of Asifit PDA, 0.5yeast PDY, 2curcuma powder PDC, 0.5 yeast and 2 curcuma powder PDM. Asifit, a tablet of food supplement for a woman in lactation, contains of 114 mg Sauropus androgynus, 20 µg vitamin B12, 15 mg vitamin B6, 2.5 mg vitamin B2, and 10 mg vitamin B1 produced commercially by Kimia Farma, Indonesia. King grass or Panicum maximum contributed around 19-20 to the overall diet in each treatment. The study was conducted according to a Complete Block design of 5 treatments and 4 replications, with 14 days for adaptation, 21 days for sampling and data recording, and 21 days post treatment was also for data recording. The dairy goats used were PE Ettawa Crossed, blocked according to their milk production. During the experiment, each of 20 goats was kept individually, provided with feed twice of 0.5kg PUFA- concentrate and 1kg fresh soybean by- product at 07.00 am and 03.00 pm. Forage was given 2kggoat at afternoon feeding. Orts of concentrate and forage were weighed in the next morning. Water was given during the afternoon time. Supplements, 3 tablets of Asifit were given orally to the respected goats in the morning feeding; whereas yeast, curcuma, or combination of both was put as topping on the mixture of PUFA- concentrate. Goats were in late lactation of 4.6 ± 0.55 months, had pre treatment milk yield of 506.4 ± 19.2 gd, body weight of 46 ± 5.34 kg. The goats were hand milked twice at 06.00 am and 02.00 pm and milk yield was recorded each morning in each individual stall. Goat assignment for each treatment was done randomly. Temperature and moisture around the housing in the upper land of Bogor, west Java, were recorded in the am and pm times daily using Hygrometer. The average temperature and moisture during the experiment were 26.94 C and 78.83, respectively. Measurement, Sampling, Nutrient and Statistical Analysis Voluntary feed intake was recorded throughout the experiment, but only the last 7 days were considered for data analysis as they were at the same days for feces collection. The samples of diets concentrate and forage and feces were oven dried 60 C for 24 hours, ground finely 1mm sieved size analyzed for dry matter DM, organic matter OM, crude protein CP by Kjeldhal method, and ether extract EE by Soxhlet method AOAC 1990. Cell wall content of acid detergent fiber ADF were determined according to Van Soest et al. 1991. Curcumin was determined by maceration method in successive steps with orange color as the final indicator Sutrisno et al., 2008. Tannin was analyzed by a modification of Folin- Ciocalteu method Harborne 1987. Calcium was determined by using Atomic Absorbance AA7000 Shimadzu Co. Serial no A 306647-00345. Phosphor was prepared by wet ashing method and quantified by using Spectrophotometer UV-200 RS UV VIS LW Scientific. Milk yield was recorded daily before, during, and after treatment period. Sampling of individual milk was collected from two consecutive milkings of the afternoon and the next morning of the last day of collection period. Milk samples were preserved in the refrigerator for later analyses of density Lactodensimeter, fat Gerber method, dry matter DM obtained by using Fleischman formula of DM = 1.311 x fat + 2.738 X 100 x density-1density; solid non fat SNF, = DM- fat; protein = fat2 +1.4. Lactose was determined according to the procedure of Teles et al. 1978. Somatic cell count was analysed by Breed method; mastitis test was done using IPB-1 reagent test Sudarwanto and Sudarnika 2008. This test was a modification of subclinical mastitis test of AMP and CMT. Scorings of mastitis were +1 starting to clot , +2 clot, and +3 glutinous. Total plate count TPC microbe was conducted by pour plate method using plate count agar PCA; Staphylococcus aureus analyzed by using Vogel Johnson agar VJA according to Veterinary Community Health Laboratory- IPB procedures. Blood samples were drawn from jugular vein into heparinized vacutainer tubes of each goat at the 35 day of the treatment. Blood parameters were analyzed for prolactin ELISA method using Spectrophotometer Human Diagnostic- USA, modified in 2007, glucose and triglyceride GOD-PAP method using Cobas 111 Roche diagnostic Ltd. Switzerland- 2010. Blood profile was determined for hemoglobin Merckotest® method using Spectrophotometer, pack cell volume or PCV Microhematocrite method, erythrocyte and leukocyte Macrohematocrite Wintrobe method. Erythrocyte part was diluted in Hayem solution and leukocyte diluted in Turk solution, each then calculated using Hemositometer Neubauer. Differentiation of leukocytes Giemsa solution coloring method contained of neutrophyle neutral granule, smooth, stick when young, segmented when old, eosinophyle big red granule, basophile big blue granule, and monocyte curvy core cell, bluish, and abundant cytoplasm. This profile was determined according to the procedures of Physiology Laboratory of Veterinary Medicine- IPB . These are some acitivities done during this experiment as shown in Figure 4.1. Data were tabulated and analyzed for variance; any differences detected were tested by Duncan Multiple Range Test DMRT in significances of P0.05 and P0.01 according to Lentner and Bishop 1986. a b c d e f Figure 4.1 Some activities done during the experiment: a PE goat; b additives; c diets; d blood; e milk; f feces samples RESULTS AND DISCUSSION Some significant differences P0.05 or P0.01 were observed either due to the treatment or the blocking of the goat, especially in milk production and milk composition. The observed treatments were affected significantly mostly in nutrient intakes and nutrient digestibilities. Diets Chemical Composition The chemical composition of diets applied in this in vivo experiment is presented in Table 4.2, showing that crude protein CP averaged about 15.00 ± 0.46; all diets were moderately higher than required for dairy goat NRC, 1981. The diets provided markedly higher dry matter DM, was around 5.07 of body weight. Comparing to diets with no or other supplements, the yeast PDY and mixture of yeast and curcuma powder PDM diets contained higher ether extract 0.33 and 0.96, respectively. Nutrient Intakes and Digestibility Intakes of dry matter DM, organic matter OM, crude protein CP, crude fiber CF, nitrogen free extract NFE, acid detergent fiber ADF, and gross energy GE were not affected significantly by dietary treatments Table 4.2. However, ether extract EE in PDM was the highest P0.05; whereas Ca and P in PDY were the highest P0.01. Consistent effects were found in feces of goat with curcuma diet PDC as it is in Table 4.3. Its lowest moisture and ash quantitatively correlated with highest dry matter, organic matter, ether extract, and crude protein. These high nutrient contents in feces were in contrast with the lower intakes of these nutrients. Apparently that they were not being digested maximally such that they were excreted in the feces. Table 4.2 Effects of feedingPUFA-diets supplemented with asifit, yeast and C. xanthorrhiza Roxb on nutrient intakes of dairy goats Intakes gd PD0 PDA PDY PDC PDM SEM P Dry matter 1228.95 1128.92 1273.07 1153.94 1239.94 60.88 NS Organic matter 1114.60 1023.76 1159.02 1054.85 1127.59 55.29 NS Crude protein 181.74 166.89 177.25 172.49 181.52 6.33 NS Ether extract 79.94 ab 73.28 a 86.67 bc 80.90 abc 91.06 c 6.79 Crude fiber 221.08 203.72 243.23 244.02 231.51 16.85 NS ADF 516.91 476.58 546.55 442.55 516.27 40.51 NS Ca 7.78 a 7.15 a 13.55 c 11.03 b 9.99 b 2.58 P 6.32b c 5.81 b 7.13 c 6.74 bc 4.67 a 0.95 PUFA- diet without supplements PD0, with 3 Asifit tablets PDA, 0.5 yeast PDY, 2 curcuma PDC, mix of 0.5 yeast and 2 curcuma PDM. P0.05; P0.01; NS: not significant. Nutrients digestibility of dry matter, organic matter, crude protein, ether extract, crude fiber, NFE, ADF, gross energy, and TDN were showed statistically the highest P0.05 in yeast diet PDY and slightly lower found in the mix diet PDM; except Ca and P were detected the highest in this diet Table 4.4. Data showed that improved nutrient digestibility as a result of increased nutrient intake that followed by increased milk yield Table 4.5. Response on intakes of yeast supplementation have been summarized as variable Desnoyers et al . 2009. There was no effects of dietary live yeast alone 1.2 10 10 cfud or in combination with mannan-oligosaccharide 32 gd on dry matter intake and milk yield of lactating Holstein Bagheri et al. 2009. In contrast, the result of yeast diet PDY intakes in this present study tended to be the highest in dry Table 4.3 Effects of feeding PUFA-diets supplemented with asifit, yeast and C. xanthorrhiza Roxb on nutrient composition of feces of dairy goats Nutrients PD0 PDA PDY PDC PDM Moisture 5.81 5.51 6.31 5.42 5.62 Dry matter 94.20 94.49 93.49 94.59 94.39 Ash 17.37 16.86 16.54 15.00 16.30 Organic matter 76.67 77.80 77.06 79.75 77.81 Ether extract 4.47 3.05 3.30 4.61 3.34 Crude protein 12.49 12.44 12.24 14.13 12.58 Crude fiber 28.03 28.36 27.56 28.08 29.55 NFE 31.84 33.78 34.06 32.77 32.62 Gross energy kcalg 3724 4002 4010 3976 3904 ADF 75.02 73.15 65.60 59.92 64.07 Ca 1.29 0.84 0.96 0.88 0.56 P 0.66 0.40 0.56 0.41 0.30 PUFA- diet without supplements PD0, with 3 Asifit tablets PDA, 0.5 yeast PDY, 2 curcuma PDC, mix of 0.5 yeast and 2 curcuma PDM. Table 4.4 Effects of feding PUFA-diets supplemented with asifit, yeast and C. xanthorrhiza Roxb on nutrient digestibility of dairy goats Digestibility PD0 PDA PDY PDC PDM SEM P Dry matter 77.09 ab 78.66 abc 84.82 c 73.77 a 83.49 bc 4.57 Organic matter 76.89 ab 78.46 abc 84.53 c 73.31 a 83.26 bc 4.62 Crude protein 79.46 b 81.00 b 85.76 b 73.80 ab 84.96 ab 4.81 Ether extract 83.30 a 89.40 b 92.13 b 81.81 a 92.05 b 4.88 Crude fiber 62.04 a 64.31 a 76.64 b 63.06 a 72.25 ab 6.43 NFE 82.82 ab 83.07 ab 87.64 b 78.24 a 87.00 b 3.79 ADF 56.53 a 60.60 ab 75.25 c 56.31 a 72.98 bc 9.13 Ca 50.45 a 70.04 b 85.37 c 74.50 b 87.83 c 14.92 P 68.76 a 82.42 b 83.79 b 80.52 b 86.03 b 6.76 TDN 72.42 a 74.07 ab 79.89 b 69.94 a 79.37 b 4.36 PUFA- diet without supplements PD0, with 3 Asifit tablets PDA, 0.5 yeast PDY, 2 curcuma PDC, mix of 0.5 yeast and 2 curcuma PDM. P0.05; P0.01. . matter and organic matters, crude fiber, and NFE. This has been supported by its high milk yield, 4 FCM, and protein weight Table 4.5. The supplementation of yeast rich in S. cereviseae, B vitamins, and amino acids has been reported to stabilize ruminal fermentation, increased rumen pH, and altered VFA levels William et al. 1991 that eventually improve milk production and milk component. Dietary yeast supplement stimulated rumen microbes growth, increased the use of N- ammonia for protein rumen microbes, and increased digestibility of fiber, increased feed intakes and nutrient supply to the gut and improved blood metabolites, eventually increased production Wallace et al. 1994. Supplementation of yeast only or in combination in this treatment did affect the goats reflecting the nature of diets, phase of lactation, feeding regime, yeast type and level, forage type and level, and environment condition as stated by Yalçın et al . 2011. In terms of tannin and curcumin contents in the rations, yeast only PDY, curcuma only PDC, or in combination with curcuma PDM seemed to affect intakes and digestibility of ether extract that were higher P0.05 than that in basal diet or Asifit diet. These supported the result of tannin and saponin sources from Yucca schidigera that increased digestibility of DM, GE, CP, and ADF Holtshausen et al. 2009. Total digestible nutrient TDN of PDY and PDM were markedly higher 79.89 and 79.37, respectively than that in other diets, such as being the lowest in PDC 69.94. The results indicated that supplementation of yeast only or in combination with curcuma was more efficient in stimulating the digestion of nutrients than that with curcuma only. To correlate with, the high nutrient contents DM, OM, CP, EE, CF, and GE in feces of the goat with curcuma diet PDC, consequently showed the opposite effects by giving the lowest digestibility P0.05 or P0.01 of these nutrients. This might be as the result of the nature of curcuma powder given on top of the concentrate that would taste bitter and smell strong that the goat avoided or by passing it in such a way that the nutrients in the diet would be decreasingly digested. In contrast, the most nutrient digestibility values both in yeast diet PDY and mix diet PDM were significantly high P0.05 or P0.01, suggesting that yeast stimulated the fermentation better either singly or in combination with curcuma. These diets were showing higher digestibilities in dry matter, and crude fiber compared to that of 1.2 10 10 cfud yeast diet in Holstein dairy cows as reported by Bagheri et al. 2009. This suggested that the yeast concentration of 1.8 10 8 cfud could be considered as the appropriate level for improving nutrient digestibility in the late lactation goat in our study. Milk Yield, Milk Component, and Milk Microbiology Milk yield and composition are shown in Table 4.5. Milk yield is calculated in gram after the application of milk density of each treatment. There were tendencies of decreasing milk yield as the time of lactation were progressing. Any supplementation did not affect significantly on milk yield pre and during treatment. In contrast, in post treatment, the milk yield was hold in better quantity with Asifit, yeast, and mix diets; whereas no supplement and curcuma diet goats were decreasing sharply P0.05 Figure 4.2. Table 4.5 Effects of feeding PUFA-diet supplemented with asifit, yeast and C. xanthorrhiza Roxb on milk yield and composition of dairy goats Variables PDO PDA PDY PDC PDM SEM P Milk yield gd Pre treatment 513 520 520 505 474 19 NS During treatment 391 524 521 372 378 78 NS Post treatment 105 a 572 b 526 b 90 a 324 ab 226 Persistency 77.1 102.2 101.7 68.6 78.4 15.4 NS Milk fat 6.83 6.65 6.73 6.60 6.15 0.26 NS 4 FCM g 548 722 729 512 477 120 NS Protein 4.81 4.73 4.76 4.70 4.48 0.13 NS Protein g 18.5 24.4 24.6 17.3 16.1 4.0 NS Fat: Protein 1.42 1.40 1.41 1.40 1.37 0.02 NS Dry matter 11.89 11.63 11.78 11.83 10.93 0.39 NS Solid non fat 5.06 4.98 5.05 5.24 4.78 0.16 NS Lactose 3.33 4.77 2.79 4.19 5.41 1.06 - Lactose g 14.89 24.60 14.22 18.53 21.06 4.33 NS Ca 0.095 0.114 0.114 0.115 0.118 0.009 - Ca g 0.42 0.59 0.58 0.51 0.46 0.07 NS P 0.114 0.129 0.145 0.173 0.152 0.022 - P g 0.510 0.665 0.739 0.765 0.592 0.110 NS CaP 0.83 0.88 0.79 0.66 0.78 0.08 NS PUFA- diet without supplements PD0, with 3 Asifit tablets PDA, 0.5 yeast PDY, 2 curcuma PDC, mix of 0.5 yeast and 2 curcuma PDM. P0.05; P0.01; NS: not significant. Figure 4.2 Milk production along the research pre, during, and post treatment However, they were different significantly P0.05 due to blocking of the goat. Blocking were supposed to reduce variability of milk yield and milk components among the goats. Contrary, after the treatments, diets with Asifit PDA and yeast PDY were significantly higher P0.05 than that in no supplement, mix, and curcuma diets. The decreasing milk yield was deeper across time in curcuma diet PDC than other treatments. This was supported by its lowest persistency 68.6. However, there was a synergistic effect when it was mixed with yeast, the milk yield was improved as high as 360. This high milk yield produced by dairy goat in late lactation fed with diets containing yeast singly or in combination with curcuma or Asifit containing Sauropus androgynus and high in vitamin B might be due to improved rumen metabolism as the result of microbial function especially S. cereviseae and vitamin B in the yeast. At the same time, bio actives tannin, curcumin, saponin, and others found in these herbs might depress unwanted microbes such as protozoa, therefore the nutrient in flow would be much more effectively converted into milk and eventually will affect milk composition as reviewed by Francis et al. 2002. Over all, milk yield of late lactation Ettawa goats in this study were about 35.54 of that in late lactation British Saanen goat Singh and Ludri 2002. Curcumin and tannin contents of these diets were decreasing average of 16.25 and 27.33 out of its pure curcuma powder 0.8 and 1.58, respectively. The lowest ether extract in curcuma diet PDC seemed to be the effect of curcumin blocking this nutrient that in some ways this was preferred for any healthy reason. Tannin contents of the diets were much lower than that in polyherbal supplements 3.69 in cross bred dairy goat as reported by Mirzaei and Prasad 2011. Curcuma supplementation, singly or in combination with yeast in this present study produced milk 19.43 than that in crossbred goat with high polyherbal combination 11.15 gd. This level of herb reduced milk yield significantly compared to that with half level in that study as reported by Mirzaei and Prasad 2011, meaning that the level in this treatment was four times higher that could have been too high for an optimal level in producing milk yield of the goat. This was in coherence with the previous level of 15 g curcumakg concentrate was the optimal level for milk production in dairy cow Sulistyowati et al . 2011. Meanwhile, Sinaga et al. 2011 reported the optimal dose 160 ppm of curcumin extract of C. domestica increased energy digestibility of ration, reduced rate of passage of the feed in the digestive tract so that its absorption improved, decreased coliform and total bacteria in feces, eventually increased body weight, but did not affect HDL level in pig. Milk fat of dairy goat fed high concentrate diet 80 with or without supplement in the present study was higher 6.59 ± 0.26 than other finding. Milk protein content averaged of 4.696 ± 0.13 equaled to 20.18 ± 0.13gd were relatively unaffected. High contents of ether extract were as the result of fatty sources incorporated in each diet. These contents were even much higher average of 8.04 compared to that in 20 extruded soybean diet 5.19 for lactating goat Schmidely et al. 2005. These results confirmed that roasted corn grain, roasted soybean meal, and corn oil did increase fat content of the diets. This could be a beneficial effect to improve low fat milk when fed to dairy goat with high concentrate. The ADF of the 0.5 yeast diet contained more than twice higher that diet with 50g Rumisacc Yalç ın et al. 2011. Populations of S. cereviseae in diets with yeast and mix diet were unusually higher than diets with no herbal supplementation. This is in the opposite with the previous result using curcuma fluid in a block supplement that caused a decrease in total fungi with the increasing level of this herb Sulistyowati et al. 2008 b . High fat contents in this present treatment were as the effect of PUFA- sources roasted corn, roasted soybean meal, and corn oil supplementation in diets fed for the goat at late lactation. This is on the contrary to other finding that high concentrate 70 with 20 extruded soybean produced low milk fat of 3.69 in dairy goat Schmidely et al. 2005, whereas, diet containing soybean oil produced higher milk fat of 5.24 in dairy goat Bouattour et al. 2008. These findings of higher milk fat out of high concentrate low fiber supplemented with these PUFA sources could be a practical solution for low fat milk producer. Supplementing fat sources to mid – late lactation goats increased milk fat content markedly +0.057 or 5.7gkg, whereas milk production was not affected Chilliard et al. 2003. This could be the reason for the opposite phenomenon that in late lactation when there is a low dilution effect due to decreasing milk yield. The dairy goat with yeast PDY diet showed second high fat percentage but had quantitatively highest 4 FCM 729gd, this was about 18.50 of that with extruded soybean. Milk protein content was higher than that in extruded soybean Schmidely et al . 2005 and soybean oil Bouattour et al. 2008, however in weight basis, the present study produced lower milk protein due to lower milk yield. Fat supplementation usually suppress milk protein content as it happened in these diets, in which mostly the milk fat and milk protein stayed relatively at the same levels. Milk lactose in the same late lactation goat but different breed was detected within the range, while the average milk fat and protein were higher than that as reported by Singh and Ludri 2002. Ratio of Ca: P in all diets were above the standard of 1.2 : 1 as required for goat according to NRC 1981. Diet with yeast and in combination with curcuma contained relatively the same higher level than other diets. This might be due to the effect of the yeast in fermenting the diet including the curcuma that made it higher in its ratio. Consequently, this supported the Ca intake that was markedly the highest 42. 69 found in the yeast diet PDY; while the curcuma and the mix diets relatively contained the same levels. Meaning that, the curcuma supplement contributed some amount of Ca higher than Asifit or no supplement diets. Over all comparison, the mix diet PDM, yeast and curcuma consistently tended to show the lowest content of milk fat, FCM, protein, fatprotein ratio, dry matter, and solid non fat SNF; however, its lactose 5.41 and Ca percentages were quantitatively the highest. Milk lactose was found the lowest 2.79 in yeast diet PDY which was unlikely since the yeast supposedly fermented carbohydrate more than other diets without yeast. Mammary Health and Hygienic Status Some indicators of mammary health and hygienic status of the goats during treatment Table 4.6 were not significantly different. Somatic cell count SCC decreased progressively in milk of goat with yeast, curcuma, and mix diet. The lowest 6.2 10 5 ml was found in the mix diet, whereas the highest 79.3 10 5 ml was detected in milk of goat with Asifit diet. One of contagious pathogens, Staph. aureus, was found the highest 222.0 cfu 10 2 ml on the contrary of the lowest SCC in milk of goat with mix supplement. This was more due to environment effects. Samples that contained S. aureus were defined as intramammary infection IMI according to NMC 1999. Milk of this treatment also contained the highest total plate count TPC of 5.05 cfu 10 3 ml. These results suggested that with the lowest SCC level in goat with mix diet, the samples were classified between 5- 6 level in Linear Score system with 45 potential loss of milk production Ingalls 2001. All milk showed SCC scores 6.2 10 5 - 7.9 10 6 cells were higher than 1 10 6 cellsml indicating an infection or mastitis Bytyqi et al. 2010. There is a different relationship between the SCC level the lowest and milk components that were also the lowest happened in samples of goat in the mix yeast and curcuma diet PDM. This was supposed in contrast of which high SCC milk shows a negative impact on milk component Sharma et al. 2011. On the other way, this lowest SCC milk showed the highest lactose content as it is otherwise stated by Harmon 1994. The antimicrobial function of curcumin in the diet curcuma only or in combination with yeast seemed to work well in suppressing the SCC. However, milk production with curcuma diet was the lowest 90 gd but improved 324 gd when combined with yeast. This herbal supplement supported the result of organic operating farm EU and USA protocol that after six months of practicing the system, there was a significant decrease in bacterial isolates found in milk of dairy cow Suriyasathaporn 2010. Concerns of high SCC milk is not only for the animals but also to human health through consuming especially the raw milk. Table 4.6 Effects of feeding PUFA-diets supplemented with asifit, yeast and C. xanthorrhiza Roxb on microbial composition and mastitis indicator of dairy goats Variable PD0 PDA PDY PDC PDM SEM P SCC cells 10 5 ml 71.8 79.3 57.1 18.5 6.2 3.27 NS Mastitis score 2.5 2.5 2.5 2.5 1.25 0.54 NS Staph. aureus cfu 10 2 ml 15.9 2.31 16.5 2.7 222.0 95.35 NS TPC cfu 10 3 ml 2.68 1.88 2.7 1.85 5.05 1.32 NS PUFA- diet without supplements PD0, 3 tablets of Asifit PDA, 0.5 yeast PDY, 2 curcuma PDC, 0.5 yeast and 2 curcuma PDM. NS: not significant. Blood Profile and Blood Metabolite Nutritional aspects determine hematological status of an animal which eventually reflects its production and health. In this study, there were no significant effects of supplementation on all blood profile Table 4.7. However, blood profile averages of Hb 9.6 gdl and PCV 25.42 were within the normal range of Hb 9- 13 gdl and PCV 23- 33. The mix diet PDM showed quantitatively the highest blood Hb and PCV, with the second highest of erythrocyte and the second lowest after the Asifit diet, while their leucocyte were at the same level. This showed that supplement rich with vitamin B in Asifit and yeast combined with herbs, Sauropus androgynus or C. xanthorrhiza Roxb will stimulate erythrocyte production and suppressed any infection indicated by lower leucocyte and lymphocyte. Differentiation of leucocytes revealed that PDM had the second lowest after the Asifit diet and the highest in neutrophyle within the range of 30- 48 and monocyte within the range of 0- 4. The blood metabolites of glucose and triglyceride in this study were higher than that in grazing goat Khaled 1999 and in goat with tea supplement Zhong et al. 2011. Prolactin in this present study was lower than that in late lactation 120- 150 days goat 4.71- 5.88 ngml Singh and Ludri, 2002. In contrast, prolactin was found the highest 4.18 ngml in diet with no supplements PD0 but its milk yield was about the same as that in yeast, curcuma, and their combination during the treatment with lower prolactin. Blood triglyceride of goat with yeast diet 37.0 mgdl was about 4.53 times higher than that in Rumisacc diet Yalçın et al. 2011. Glucose was the highest in mix diet PDM, while blood prolactin in goat with this diet was the lowest. However, this milk yield was plunging markedly P0.05 even though its prolactin was that high after the treatment. Table 4.7 Effects of feeding PUFA-diets supplemented with asifit, yeast and C. xanthorrhiza Roxb on blood parameters of dairy goats Variable PD0 PDA PDY PDC PDM SEM P Hemoglobin gdl 9.65 9.17 9.89 9.36 9.97 0.34 NS PCV 25.89 24.63 26.48 22.81 27.30 1.75 NS Erythrocyte 10 6 mm 3 16.60 19.57 17.67 18.91 19.36 1.26 NS Leucocyte 10 3 mm 3 14.45 12.53 13.19 15.19 12.53 1.19 NS Differentiations of Leucocyte Lymphocyte 52.75 39.00 44.50 42.50 40.67 5.36 NS Neutrophyle 38.00 50.00 49.50 48.25 51.00 5.32 NS Monocyte 2.75 4.00 2.75 2.50 4.33 0.84 NS Eosinophyle 6.50 7.00 3.25 6.75 4.00 1.74 NS Basophyle - - - - - Glucose mgdl 53.75 50.25 47.25 54.75 58.00 4.15 NS Triglyceride mgdl 25.50 25.00 37.00 32.50 28.75 5.04 NS Prolactin ngml 4.18 3.58 3.58 2.99 1.64 0.07 NS PUFA- diet without supplements PD0, with 3 Asifit tablets PDA, 0.5 yeast PDY, 2 curcuma PDC, mix of 0.5 yeast and 2 curcuma PDM; NS: not significant. Blood glucose 58 mg100 ml in goat with the mix diet was exactly the same level as in 56gd XP yeast diet in dairy cow Hristov et al. 2010, but was slightly higher than that in dairy goat Singh and Ludri 2002. Yeast diet PDY, in contrast, showed the lowest blood glucose as it was reflected by the lowest milk lactose and highest triglyceride. The PUFA diet supplemented with PDM mix of yeast and curcuma improved the physiological status of the goats showed by the high metabolite yield of glucose. Therefore, prolactin only, without support of a good amount of milk precursor will not improve milk yield. On the other hand, yeast contained S. cereviseae that was important during fermentation in rumen, while curcuma powder contained curcumin acting as antimicrobial, such as suppressing protozoa in the rumen in such a way that nutrient metabolism process was improved, eventually the blood metabolites, milk yield, and milk composition will be improved as well. Therefore, a synergistic function between both supplements gave a positive impact in the blood values of the goats. CONCLUSION A mix supplement of 0.5 yeast and 2 curcuma powder is considered good since it showed a better recovery in milk yield post treatment with progressing lactation in dairy goat. Besides, it showes some tendencies of lower milk fat, 4 FCM, and SCC; higher lactose and Ca percentages; higher Hb, PCV, and glucose in blood; higher ether extract intakes and Ca digestibility, supported by relatively high nutrient digestibility in most nutrients.

4. FATTY ACID QUALITY OF MILK OF LATE LACTATION

DAIRY GOAT FED PUFA-DIET SUPPLEMENTED WITH YEAST AND C. xanthorrhiza Roxb ABSTRACT 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 quality of goat fed with this diet showed high in LCFA and MUFA; while it was low in SCFA, n6n3 ratio, and atherogenicity index. These qualities were optimally considered good in terms of healthier product. Therefore, the PUFA- diet with 0.5 yeast and 2 curcuma C. xanthorrhiza Roxb was a reasonable choice to be applied for dairy goat. Key words: curcuma, dairy goat, milk fatty acid, PUFA-diet, yeast INTRODUCTION Milk fatty acid has been an increasing concern for healthy reason. Diet with high concentrate mostly increase milk production, however, it also decrease milk fat and changed milk fatty acid, such as C 10, C12, C18:1t, C18:1c, C18:2n6c Calderon et al. 1984; Ledoux et al. 2002. Different from milk of cow that contains higher LCFA; milk dairy goat is higher in SCFA, and medium chain fatty acid MCFA, according to Bouattour et al. 2008. There have been some nutrient efforts to modify this fatty acid content. Some of these efforts that can be applied in dairy goat were providing diet containing PUFA supplemented with yeast and curcuma. Flaxseed containing concentrate, treated or untreated with formaldehyde, showed moderate amount of C18:1 and high in C18:3. Whereas, in sun flower containing concentrate, both treated and untreated, C18:2 was found being the highest Petit 2003. This suggested that formaldehyde was not effective enough to protect polyunsaturated fatty acids against ruminal biohydrogenation. Polyunsaturated fatty acid PUFA supplementation in concentrate is incompletely biohydrogenised later in the biosystem of ruminants which eventually affects fatty acid in milk Whitlock et al. 2002; Schmidely et al. 2005. Therefore, it needs an effort to overcome this process, such as trough fat supplementation Chilliard et al. 2003; addition of extruded soybean decreased SCFA and LCFA Whitlock et al. 2002 and Schmidely et al. 2005; also, addition of flaxseed products reduced n6n3 in milk of dairy cow Cortes et al. 2010. Feeding concentrate containing yeast and curcuma with different fatty acid sources palm oil showed optimal levels in PUFA, ratio of PUFAsaturated PS, and n6n3 designated for dairy cows Sulistyowati et al. 2010 c . Yeast supplementation was reported to affect fatty acid content in milk. Addition of 50gd RumiSacc containing 33.01 saturated fatty acid, 26.42 5. PUFA, and 66.99 UFA did not affect the 18:3 n-6, but did increase 18:3 n- 3 in milk of dairy cow Yalçın et al. 2011. Curcuma, a medicinal herb, contains bioactives of curcuminoids curcumin, demethoxycurcumin, and bisdemethoxycurcumin. It has several biological activities, such as antiinflammatory, antioxidant, and antimicrobial Jayaprakasha et al . 2002, antifungal and antibiofilm Rukayadi et al. 2008, also hypocholeretic. Liquid extract of C. xanthorrhiza Roxb in Tabut block was able to decrease fat content in milk of lactating Bali cow Sulistyowati et al. 2001. Sinaga 2011 found that curcumin 160 ppm in C. domestica did not affect HDL level in pig. Other herbal source, tea catechin inclusion was reported to decrease markedly in low-density lipoprotein cholesterol and triglyceride in blood of goat Zhong et al. 2011. Based on these data, an experiment has been conducted to investigate the effects of different additives, yeast, curcuma and their combination were added in concentrate containing PUFA sources roasted ground corn, roasted soybean meal, and corn oil on milk fatty acid in milk of dairy goat. MATERIALS AND METHODS Preparation of Yeast, Curcuma, and PUFA- Diet Yeast additive was prepared from several ingredients based on a modification of the procedure of Pusbangtepa 1981. The ingredients were rice flour, cassava tuber, sugar, garlic, local yeast Bengkulu, lemon juice, water, and A. galanga Sw. This made yeast containing 3.6 10 7 cfug was then ready to be added as dried supplement. Curcuma powder was made of C. xanthorrhiza Roxb tuber, sliced, sun dried, oven dried, grounded and refined. The curcuma powder was about 26.6 ww out of fresh C. xanthorrhiza Roxb. Bioative contents of this curcuma powder were curcumin 0.8 and tannin 1.58. Diet containing PUFA – concentrate from the previous experiment was then combined with soybean by-product used to be provided on the farm was formulated for 30 kg lactating dairy goat with 1kg of milk production NRC 1981 can be seen on Table 5.1. The ground corn was half roasted, while the soy bean meal was all roasted in 80° C until turned light brown. These PUFA sources contained of 50.35, 34.69, and 46.46 out of total FAME fatty acid methyl esters, for roasted soy bean meal, roasted ground corn, and corn oil were intended as containing, respectively. The cassava meal was prepared from the fresh tubers, sliced, sun dried, then grinded as powder. Soybean by product was in the form of the skin of soybean after an extrusion from boiling it made in some small scale tofu industries. Mixing of the ingredients was started from the smallest portion, manually as homogenized as possible. Each treatment was prepared with the addition of the respective supplement. Treatments, Dairy Goats, and Diet Analysis There were five treatments PD0: PUFA- diet with no additive, as negative control; PDA: PUFA- diet with 3 Asifit tablets, as positive control; PDY: PUFA- diet with 0.5 yeast; PDC: PUFA- diet with 2 curcuma powder; and PDM: PUFA- diet with a mixture of 0.5 yeast and 2 curcuma powder applied in 20 late lactation dairy goats. The goats were Ettawa crossed breed, in late lactation 4.6 ± 0.55 months, with body weight of 46 ± 5.34 kg, hand milked twice at 06.00 am and 03.00 pm and recorded in each individual stall. Asifit, a tablet of supplement for a woman in lactation, contains of 114 mg Sauropus androgynus , 20 µg vitamin B12, 15 mg vitamin B6, 2.5 mg vitamin B2, and 10 mg vitamin B1 produced commercially by Kimia Farma, Indonesia. King grass or Panicum maximum contributed around 19-20 to the overall diet in each treatment. Nutrient analyses of dry matter DM, organic matter OM, crude protein CP, crude fiber CF, and ether extract EE were determined according to AOAC 1990. While, NFE was calculated as 100 - moisture + ash + EE + CP + CF. The content of NDF and ADF were analyzed by the method of Van Soest 1990. Minerals of Ca λ 422.7 nm was analyzed using Atomic Absorbance Spectrophotometer AAS AA7000 Shimadzu Co. Serial no A 306647-00345; while P λ = 660 nm was analyzed using wet ashing method and quantified using Spectrophotometer UV-200 RS UV VIS LW Scientific. Bioactive analyses of curcumin was determined by maceration method with the final product of crystal with orange- brown color as curcuminoid, in which curcumin was part of it Sutrisno et al. 2008. Tannin was analyzed by polyphenol redox method then detected using Spectrophotometer λ 725 nm. Table 5.1 Ingredients and nutrient composition of PUFA-diet containing asifit, yeast and C. xanthorrhiza Roxb for dairy goats Ingredients PD0 PDA PDY PDC PDM Rice bran 18.42 18.42 18.42 18.42 18.42 Ground corn 15.79 15.79 15.79 15.79 15.79 Soy bean meal 7.89 7.89 7.89 7.89 7.89 Cassava meal 7.89 7.89 7.89 7.89 7.89 Soy bean by-product 27.49 27.49 27.49 27.49 27.49 Corn oil 2.11 2.11 2.11 2.11 2.11 Mineral 0.53 0.53 0.53 0.53 0.53 King grass KG 19.88 19.88 19.88 19.88 19.88 Asifit - 0.22 - - - Yeast - - 0.5 - 0.5 Curcuma powder - - - 2.0 2.0 PUFA- diet without supplements PD0, with 3 Asifit tablets PDA, 0.5 yeast PDY, 2 curcuma PDC, mix of 0.5 yeast and 2 curcuma PDM. Sample Collection, Data and Statistical Analysis Sampling and data recording were based on the length of the research that was divided into 14 days for adaptation, 21 days for, and 21 days post treatment. The application of the experimental diets was in a Randomized Complete Block design of 5 treatments and 4 replications. Data were tabulated and statistically analyzed in Anova and any differences were detected using DMRT Lentner and Bishop 1986. Each of the 20 goats was kept individually, provided with feed twice of 0.5kg PUFA- concentrate and 1kg of fresh soybean by-product at 06.00 am and 03.00 pm. Forage was given 2kggoat on afternoon feeding. Orts of concentrate and forage were weighed in the next morning. Water was given during the afternoon feeding time. The 3 tablets of Asifit were given orally to the respected goats in the morning feeding; whereas yeast, curcuma, or combination of both was put as topping on the mixture of PUFA- concentrate. As the Asifit tablets were given orally, therefore the analyses of this diet was using the same result as the control diet PD0. Goats were hand milked twice 06.00 am and 03.00 pm and recorded in each individual stall. Milk yield was recorded daily before, during, and post treatment. Sampling of individual milk was collected from two consecutive milkings of the afternoon and the next morning of the last day of collection period. PUFA Analyses, Calculation, and Environment Milk fatty acids were analyzed after the extraction of milk fat samples and methylation yielding fatty acid metil ester FAME. The same procedures were also applied for diets and feces. Separation and quantification of the FAME were conducted by using gas chromatography GC- FID, 2010 Plus, Shimadzu, Japan. Groups of fatty acids were calculated as follows: total SCFA short chain fatty acid: C 10, MCFA medium chain fatty acid: C12- C16, LCFA long chain fatty acid: CC16, MUFA mono unsaturated fatty acid: C with one double bond, PUFA poly unsaturated fatty acid, saturated and unsaturated fat, ratio PUFAsaturated fat, ratio n6n3: linoleic acid + arachidonic acidlinolenic acid Schmidely et al. 2005, and atherogeneicity: atherogenicity index = C12 + 4C14 + C16total unsaturated fat Ulbricht and Southgate 1991. Environment temperature and humidity around Cordero farm in the upper land of Bogor, west Java, were recorded at 6 am and 3 pm daily, using Haar-Synt Hygrometer made in Germany. The averages of temperature and humidity during the experiment were 26.94 C and 78.83, respectively. RESULTS AND DISCUSSION Diets Chemical Composition The average of crude protein Table 5.2 of all diets was around 15.00 ± 0.46; these diets were moderately higher than that of required for dairy goat, 12 NRC 1981. The DM of diets provided around 5.07 of body weight; this was higher than it is supposedly for DM required for 30 kg dairy goat and 1kg of milk production. The PDY and PDM diets similarly contained higher ether extract than others; these were as the result of incorporation of fatty sources in all diets. These contents were even much higher average of 8.04 compared to that of 20 extruded soybean diet 5.19 for lactating goat Schmidely et al. 2005. These results confirmed that the fatty acid sources roasted corn grain, roasted soybean meal, and corn oil increased fat content of the diets in this present experiment. This finding could be a beneficial effect to dairy goat with high concentrate so that it will improve low fat milk that used to be with this level of concentrate. The 0.5 yeast diet PDY contained more than twice higher in ADF than that of diet with 50g Rumisacc, according to Yalçın et al. 2011. Saccharomyces cereviseae populations in diets with yeast PDY and mix diet PDM were higher than other diets; however, the curcuma in the mix diet was affecting the population by lowering it. Curcumin and tannin in these diets were decreasing in the average of 16.25 and 27.33, respectively, compared to their intial source in C. xanthorrhiza Roxb powder. The curcuma diet PDC showed the lowest ether extract, suggesting that curcumin blocked this nutrient which is preferred for healthy reason. Tannin contents in all diets were much lower 13.3 out of 3.69 polyherbal supplements in cross bred dairy goat Mirzaei and Prasad 2011. Table 5.2 Nutrient composition of PUFA-diet containing asifit, yeast and C. xanthorrhiza Roxb for dairy goats Nutrients PD0-A PDY PDC PDM KG Dry matter 94.13 93.96 93.72 94.06 20.10 Ash 8.48 7.96 7.82 8.21 11.65 Crude protein 15.27 14.11 15.33 15.04 13.42 Ether extract 7.66 8.14 7.96 8.77 3.22 Crude fiber 13.30 14.57 18.60 14.51 31.32 NFE 49.12 49.18 44.00 47.60 32.64 ADF 29.07 29.54 48.18 29.35 78.99 Gross energy Mcalkg 3.84 3.86 3.83 3.90 3.71 Ca 0.61 1.20 1.02 0.84 0.7 P 0.47 0.53 0.57 0.29 0.13 CaP 1.30 2.26 1.79 2.90 0.64 Tannin 0.487 0.492 0.491 0.491 0.491 Curcumin - - 0.13 0.11 - S. cereviseae 10 6 cfug 1.6 7.3 1.2 4.0 - PD0:PUFA- diet with no suppl.; PDA= PD0-A; PDY: PUFA- diet with 0.5yeast; PDC: PUFA- diet with 2curcuma; PDM: PUFA- diet with 0.5yeast + 2curcuma. KG: King grass. Ratio of Ca to P in all diets were above 1.2 : 1 as required for dairy goat according to NRC 1981. The PDY and PDM showed higher Ca to P ratios of 2.26 and 2.90, respectively. This might be as the effect of yeast fermenting these diets in such a way that Ca within these diets became more available; then when combined with curcuma, it provided more available Ca. Fatty Acid Profile of PUFA-Diets Noticeable fatty acids Table 5.3, such as in PD0-A was found three times higher than others in stearate C18:0, 13 times smaller in PS ratio, eight times smaller than that of PDM in oleate C18:1n9t, three times smaller than that of PDM in MCFA, 16 times smaller than that of PDM in PUFA, but 2.3 times higher in n6n3 ratio than those of the three other diets. As of PDM, there were some marked higher results, such as in oleate C18:1n9t, linoleate C18:2n6c, linolenate C18: 3n3, EPA, total fatty acid, total MCFA, and total PUFA. On the other hand, PDY was high in total LCFA, total MUFA, and total saturated fat; while PDC was high in total unsaturated and US ratio, but low in atherogeneicity index. However, the three treated PUFA- diets PDM, PDC, and PDY were similarly had higher PS and lower n6n3 than that of PD0-A. These PS ratios were lower than the previous PUFA-concentrate containing palm oil and roasted ground corn 2.99- 3.07 and the n6n3 ratios were much higher in this present data, compared to 1.5- 1.74 reported by Sulistyowati et al 2010 b . However these present data were within the range of 0.88- 1.69 in concentrate with saturated hydrogenated fat and unsaturated ca-soap fat, respectively Harvatine and Allen 2006. These suggested that the type of fat incorporated in concentrate would determine the type of fatty acid yielded. Based on these fatty acid quality, it seemed that PUFA-diet with a mixture of 0.5 yeast and 2 curcuma powder PDM was considered the most effective diet then followed by PDC and PDY. Suggesting that yeast additive worked in fermenting nutrients, at the same time the curcuma controlled the fatty acid contents in diets containing PUFA and Allen, 2006. These suggested that the type of fat incorporated in concentrate would determine the type of fatty acid yielded. Based on these fatty acid quality, it seemed that PUFA-diet with a mixture of 0.5yeast and 2curcuma powder PDM is considered the most effective diet then followed by PDC and PDY. Suggesting that yeast additive worked in fermenting nutrients, at the same time the curcuma controlled the fatty acid contents in diets containing PUFA. Table 5.3 Total fatty acid contents in PUFA-diets supplemented with Asifit, yeast and C. xanthorrhiza Roxb for dairy goat Fatty acid of total fat PD0-A PDY PDC PDM Capric Acid, C10:0 nd nd nd 0.01 Undecanoic acid, C11:0 nd nd nd 0.02 Lauric Acid, C12:0 0.23 0.21 0.17 0.23 Tridecanoic Acid, C13:0 0.02 nd nd nd Myristic Acid, C14:0 0.29 0.18 0.15 0.21 Pentadecanoic acid, C15:0 0.39 0.05 0.04 0.06 Palmitic Acid, C16:0 3.9 10.92 8.94 12.68 Palmitoleic Acid, C16:1 0.4 0.08 0.06 0.09 Heptadecanoic Acid, C17:0 0.38 0.1 0.08 0.12 Stearic Acid, C18:0 9.07 3.05 2.3 3.48 Elaidic Acid, C18:1n9t nd 0.06 0.04 nd Oleic Acid, C18:1n9c 2.21 15.54 11.7 17.37 Linolelaidic Acid, C18:2n9t 0.07 nd nd 0.05 Linoleic Acid, C18:2n6c 0.92 13.42 10.8 14.85 Linolenic Acid, C18:3n3 0.07 2.31 1.94 2.55 Arachidic Acid, C20:0 0.29 0.66 0.47 0.71 Cis-11-Eicosenoic Acid, C20:1 nd 11 0.08 0.12 Cis-11,14-Eicosedienoic Acid, C20:2 nd 0.07 0.09 0.1 Cis-5,8,11,14,17-Eicosapentaenoic acid,C20:5n3 nd 0.02 nd 0.4 Heneicosanoic Acid, C12, C21:0 0.02 0.05 0.03 0.06 Behenic Acid, C22:0 0.46 0.59 0.44 0.67 Tricosanoic Acid, C23:0 0.04 0.11 0.08 0.12 Lignoceric Acid, C24:0 0.3 0.39 0.27 0.45 Nervonic acid, C24:1 nd nd nd 0.01 Total fatty acid 19.17 48.02 37.74 54.16 Total SCFA nd nd nd 0.01 Total MCFA 4.83 11.36 9.30 13.18 Total LCFA 13.83 47.37 28.32 40.7 Total MUFA 2.61 26.68 11.88 17.59 Total PUFAP 1.06 15.82 12.83 17.59 Total Saturated fat S 15.39 16.31 12.97 18.82 Total Unsaturated U 84.61 83.69 87.03 81.18 Rasio PS 0.07 0.97 0.99 0.93 Rasio US 5.50 5.13 6.71 4.31 n6n3 13.14 5.83 5.57 5.86 Atherogenicity 1.44 1.06 0.87 1.23 PUFA- diet without supplements PD0, with 3 Asifit tablets PDA, 0.5 yeast PDY, 2 curcuma PDC, mix of 0.5 yeast and 2 curcuma PDM; nd: not detected.