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

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 PUFAsaturated PS, and n6n3 designated for dairy cows Sulistyowati et al. 2010 b . 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,10.5of sugar, 10g of garlic, 20.5of 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 10 7 cfug. 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 ww 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 20gkg of PUFA- concentrate, mixed thoroughly. Steps in preparing the curcuma powder were presented in Figure 2.2. PUFA- Concentrate Preparation The concentrate was 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 corn 1 30 30 30 30 Cassava meal 15 15 15 15 Soy bean meal 2 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 Requirement 3 Dry matter 84.85 2.7 TDN 69.10 70.40 DE Mcalkg 3.21 3.10 NEL Mcalkg 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 0.5yeast; PCC: PUFA - concentrate with no yeast and 2curcuma; PCM:PUFA -concentrate with 0.5yeast and 2curcuma; 1 half 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 2curcuma; 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 P0.05 or P0.01 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 CC16 72.83 69.66 60.81 Total MUFA 23.92 15.43 22.06 Total PUFAP 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 PS 4.10 4.12 2.46 Ratio US 7.83 7.18 6.09 n6n3 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 FeCl 3. 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 10 6 , then 0.1 ml was taken appropriately and inoculated onto 10 ml NA Nutrient Agar plate, made in double. Plates were incubated unaerobic at 40 C 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 C16, mono unsaturated, poly unsaturated, saturated, unsaturated, ratio of PUFAsaturated, and unsaturatedsaturated fatty acid. Fatty acid ratio of n-6n-3 was calculated using this formula Schmidely et al. 2005 : n6n3 = linoleic acid + arachidonic acidlinolenic 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.