Biol. Chem. Research. 43 Isolation of Saccharomyces Spp from Manure of Beef Bali Cattle as a Probiotics Properties and has CMC-ase Activity to Improve Nurient Quality of Rice Bran.

I solation of……………………………….Rice Bran Candraw ati, et al., 20 14 Calculations The data w ere used t o calculat e AM E value according to t he follow ing form ulat e M ust afa et al., 2004: AM E apparent m et abolizable energy = IE – FE. Where IE = ingest ed energy; FE = fecal energy voided by the fed birds. St atistical Analysis All dat a w ere subject ed t o a one-w ay analysis of variance t est St eel and Torrie, 1989. St at ist ical significances among t reat m ent means w ere det erm ined by m et hod of New M ult iple Range Test of Duncan when t he F value w as significant at 5 level RESULTS Resilience isolat e Saccharomyces spp. t he low pH is one of t he charact erist ics required t o be met by a candidat e or a probiot ic that could be developed into a pot ential probiotic. On t his t est , t he m edium pH w as adjust ed t o 1.5, 3.0, 4.5, and 6.0 by using HCl hydrochloric acid, because HCl has charact erist ics similar t o stom ach acid. Resist ant to highly acidic nature needs t o be ow ned by t he candidat e probiotic, because t he applicat ion lat er, t his probiot ic candidat e must pass very acidic st omach conditions, before reaching t he colon. Observat ions as present ed in Table 1. In this st udy, as m any as 10 isolat es Saccharomyces spp . isolat ed from catt le feces w as t est ed resist ance t o various pH, at pH 1.5, 3.0, 4.5, and 6. Two isolat es w ere unable t o grow at pH 1.5, w hich isolates t he S-1 and S-2. Saccharomyces spp .S-1 isolat es even sim ply could not grow at all pH t reat m ents given. Only 8 isolat es w ere able t o grow w ell, nam ely Saccharomyces spp.S-4 t o S-10 isolat es. There is a t endency of t he higher pH, especially at pH 6, most of t he isolat es has decreased t he number of colony life. Saccharomyces spp colonies grow w ell at pH 3, even isolat e S-3 and S-5 Saccharomyces spp colony number reached 5.531 and 5.093 log colony per gram . Table 1. The number of colony isolates Sa ccharomyces spp Log colony g at various pH. Code Isolat e pH 1.5 3 4.5 6 S 1 - - - - S 2 - 3.98 3.88 3.32 S 3 3.45 5.53 4.41 4.13 S 4 4.33 3.65 3.58 4.56 S 5 4.12 5.09 4.99 4.12 S 6 4.19 4.35 4.36 4.40 S 7 4.29 4.42 3.99 3.58 S 8 4.06 3.617 4.318 3.86 S 9 4.31 4.41 3.95 3.92 S 10 4.16 4.33 3.97 3.87 Descript ions: S-1 s d S-10 is a Saccharomyces spp isolat es w ere isolat ed from manure of beef Bali cat tle

J. Biol. Chem. Research. 43

Vol. 3 1, No. 1: 3 9-52 20 14 I solation of……………………………….Rice Bran Candraw ati, et al., 20 14 In the t est of resist ance t o bile salt s as seen in Table 2 show s, that the t en isolat es w ere able to survive and grow on a medium cont aining bile salt s at concent rat ions of 0.2 m M and 0.4 m M , whereas at a concent rat ion of 0.6 m M no isolat es can survive. M icrobial isolat es resist ance t o bile salt s is used t o assess t he abilit y t o survive t he digest ive t ract isolat es cont ained bile salt s on the surface of t he int est ine. Probiot ics w ill be dealing w ith the environment in t he sm all int est ine, in which t here are bile or bile salt s are rem oved by t he liver t hrough t he gall bladder, aft er successfully passing the acidic condit ions in the st om ach. Therefore, in t he process of developing new probiotic, or a new probiotic candidat e m ust be able t o pass t he t est of resist ance t o bile or bile salt s w ere perform ed in vit ro. Based on t he nat ure of t he resist ance show n by som e isolat es, st rai-indicat es t hat t he st rain has t he pot ential to be developed int o a pot ent ial probiot ic. Table 2. Ability of yeast Sa ccharomyces spp isolates w ere resistant living in bile salts. Isolat e Bile salt concent rat ion absorbance 0.2 mM 0.4 mM 0.6 mM S 1 + 0.244 + 0.169 - 0.067 S 2 + 0.248 + 0.168 - 0.059 S 3 + 0.253 + 0.163 - 0.081 S 4 + 0.252 + 0.158 - 0.064 S 5 + 0.243 + 0.166 - 0.073 S 6 + 0.224 + 0.144 - 0.036 S 7 + 0.192 + 0.133 - 0.031 S 8 + 0.145 + 0.151 - 0.051 S 9 + 0.126 + 0.132 - 0.044 S 10 + 0.138 + 0.149 - 0.035 Descript ion: - : A 0.1 not bear bile salt + : A 0.1 - 0.5 a bile salt -resist ant + + : A 0.5 - 1.0 bile salt -resist ant + + + : A 1.0 highly resist ant of bile salt s S-1 s d S-10 is a Saccharomyces spp isolat es were isolat ed from m anure of beef Bali cat tle CM C-ase t est is t o t est t he abilit y of t he yeast Saccharomyces spp isolat es in degrading crude fiber. It can be m easured from t he result ing clear zone diam et er Table 3. From t he result s of t his st udy w ere only yeast Saccharomyces spp isolat es S-5, S-6, S-7, S-8, and the yeast Saccharomyces spp.S-9 who has the abilit y t o digest fiber. Looks yeast Saccharomyces spp isolat es S-6 has the most wide clear zone, while the S-8 has a clear zone at least .

J. Biol. Chem. Research. 44

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