Effects of Replacing Concentrate with Soybean Curd Residue Silage on Ruminal Characteristics, Plasma Leucine and Glucose Turnover Rates in Sheep

Transcription

1 Journl of Animl Science Advnces Effects of Replcing Concentrte with Soyen Curd Residue Silge on Ruminl Chrcteristics, Plsm Leucine nd Glucose Turnover Rtes in Sheep Hrjnti D. W., Sugwr Y., Al-Mmun M. nd Sno H. J Anim Sci Adv 2012, 2(4): Online version is ville on:

2 ISSN: HARJANTI ET AL. Originl Article Effects of Replcing Concentrte with Soyen Curd Residue Silge on Ruminl Chrcteristics, Plsm Leucine nd Glucose Turnover Rtes in Sheep Hrjnti D. W., Sugwr Y., Al-Mmun M. nd Sno H. Deprtment of Animl Sciences, Fculty of Agriculture, Iwte University, Moriok , Jpn Astrct The effects of replcing concentrte with soyen curd residue silge on the plsm leucine (Leu) nd glucose turnover rtes (LeuTR nd GluTR) of sheep were evluted using n isotope dilution method of [1-13 C]Leu nd [U- 13 C]glucose. The ruminl chrcteristics nd nitrogen (N) lnce of the sheep were lso determined. The sheep were fed two diets in crossover experiment for 21 dys. The first diet consisted of 80% mixed hy nd 20% commercil concentrte (CON diet), nd the second diet consisted of 80% mixed hy nd 20% soyen curd residue silge (SCRS diet), on dry mtter sis for oth diets. The primed-continuous infusion method of [1-13 C]Leu nd [U- 13 C]glucose ws performed on dy 21. The N sorption, N retention, nd N digestiility did not differ etween the diets. Furthermore, the ruminl totl voltile ftty cid concentrtion did not differ, ut the molr concentrtion of propionte ws significntly higher (P=0.02) in the sheep fed the SCRS diet thn the CON diet. The plsm LeuTR nd GluTR were comprle etween the diets. The results showed tht soyen curd residue silge could e comprle with commercil concentrte on ruminl fermenttion s well s on the plsm Leu nd glucose kinetics. Thus, soyen curd residue silge could e used to replce commercil concentrte in the diet of sheep. Key words: food y-product, glucose kinetic, isotope dilution, protein kinetic, wether * Corresponding Author: Deprtment of Animl Sciences, Fculty of Agriculture, Iwte University, Ued , Moriok , Jpn; sno@iwte-u. c.jp Received on: 26 Fe 2012 Revised on: 13 Mr 2012 Accepted on: 25 Mr 2012 Online Pulished on: 14 Apr J. Anim. Sci. Adv., 2012, 2(4):

3 EFFECTS OF REPLACING CONCENTRATE WITH SOYBEAN CURD RESIDUE SILAGE ON RUMINAL Introduction Economic nd environmentl concerns prompt investigtion into the possiility of using the orgnic resources generted y the food industry s lterntive niml feeds. Soyen curd residue (SCR), y-product of tofu, soymilk nd soy protein mnufcturing, is high in protein content; however, ecuse SCR is considered n industril wste, it is often incinerted due to its rpid rte of deteriortion. Ensiling SCR (soyen curd residue silge; SCRS) could prolong its shelf life nd incresed its nutrient composition nd digestiility (Cho et l., 2007). An in vitro study on the nutritionl properties of SCRS showed tht the fermenttion process lso enhnced ntioxidnt ctivity (Rshd et l., 2001), nd previous studies (Oltjen & Putnm, 1966; Clifford & Tillmn, 1968; Prior, 1976; Knus et l., 2002) hve reported incresed nitrogen (N) sorption nd N retention when soy protein-sed diets were fed to steers nd sheep. Inclusion of SCRS in the diet of sheep hs een shown to increse the glucogenic propionte concentrtion in the rumen without ny negtive effects on ruminl fermenttion (Xu et l., 2001). Nevertheless, reserch regrding the use of SCRS s ruminnt feed on the N lnce nd the kinetics of plsm mino cid (AA) nd glucose hs een limited. Thus, dditionl studies should e needed to promote the utiliztion of this yproduct s ruminnt feed. We hypothesized tht SCRS cn replce commercil concentrte in the diet of sheep without ny dverse effects on whole-ody N nd glucose metolism due to its high solule N nd fermentle crohydrte contents. Improved utiliztion of residue generted y the food industry such s SCRS could reduce feed cost nd environmentl prolem. The ojective of this study ws to determine the effects of replcing commercil concentrte with SCRS on the plsm leucine (Leu) nd glucose turnover rtes (LeuTR nd GluTR), using n isotope-dilution method with [1-13 C]Leu nd [U- 13 C]glucose. The N lnce, ruminl chrcteristics nd lood metolites were lso nlyzed. Mterils nd Methods Animls nd diets All of the niml hndling, including lood smpling, ws conducted ccording to the guidelines estlished y the Animl Cre Committee of Iwte University. Four crossred (Suffolk x Corriedle) wether, ll of pproximtely 4 yers of ge nd initilly of 54 ± 4 kg of ody weight were used. This experiment used crossover design: 21-dy period tht consisted of 14 dys of dietry dpttion nd 7 dys of smple collection. Throughout the dpttion period, the sheep were kept in individul pens nd fed two dietry tretments. The first diet consisted of 80% mixed hy (orchrdgrss nd reed cnrygrss) nd 20% commercil concentrte (CON diet), nd the second diet consisted of 80% mixed hy nd 20% soyen curd residue silge (SCRS diet), on dry mtter (DM) sis. The SCRS contined 15% eet pulp. The chemicl composition of mixed hy, SCRS nd commercil concentrte re listed in Tle 1. The diets were estimted to e isonitrogenous nd the metolizle energy intkes ssumed from the Ntionl Reserch Council (1985) were slightly ove the mintennce level for oth diets, s shown in Tle 2. The sheep were fed twice dily t nd hours nd hd d liitum ccess to wter. The SCRS ws purchsed from food compny (Hirkw Food Co. Ltd., Iwte, Jpn), plced into 90 L polyvinyl gs, compcted, nd seled tightly. After 15 dys of ensiling, the continer ws opened nd the silge ws fed to the nimls s reported previously (Amh et l., 1995). On dy 15, the sheep were moved to individul metolic cges in controlledenvironment room, with n ir temperture of 23⁰C, reltive humidity of 70%, nd lighting from to hours. Smple collection Before strting the feeding experiment, smples of the feed were collected nd dried in forced-ir oven t 60⁰C for 48 hours to determine the chemicl composition nd to formulte the dietry tretments. Offered feed nd refused feed (if ny) were smpled nd dried using the sme procedure for the chemicl nlysis. From dy 16 to dy 20, urine nd feces smples were collected t 24-hour intervls using 362 J. Anim. Sci. Adv., 2012, 2(4):

4 HARJANTI ET AL. 3 mm plstic screen seprtor. The smples were collected once dily t hours. The urine ws collected from ech sheep in ucket contining 50 ml of 6 N H 2 SO 4, nd the volume ws recorded; 50 ml of the urine ws stored t -30⁰C until nlysis. Feces smples were dried in forced-ir oven t 60⁰C for 48 hours nd were then pssed through 1 mm mesh nd stored t room temperture until nlysis. Tle 1: Chemicl compositions of the mixed hy, soyen curd residue silge nd commercil concentrte Mixed hy Soyen curd residue silge Commercil concentrte DM (g/kg) CP (g/kg DM) EE (g/kg DM) NDF (g/kg DM) ME (Mcl/kg DM) c 2.68 Soyen curd residue silge contined 15% eet pulp (Hirkw Food Co. Ltd., Jpn) Commercil concentrte: formul feed ( α-beef mde y Chuu Feed Co. Ltd., Jpn) c Clculted s the proportion of soyen curd residue nd eet pulp (85:15) in the silge ccording to MAFF (1995). DM: dry mtter CP: crude protein EE: ether extrct NDF: neutrl detergent fier ME: metolizle energy Tle 2: Diet formultion nd intkes of crude protein (CP) nd metolizle energy (ME) of the dietry tretments Tretment CON diet SCRS diet Mixed hy (g. kg BW d -1 ) Commercil concentrte (g. kg BW d -1 ) Soyen curd residue silge (g. kg BW d -1 ) CP intke (g. kg BW d -1 ) ME intke (Mcl. kg BW d -1 ) Tretment: mixed hy (orchrdgrss nd reed cnnrygrss) s the sl diet which is supplemented with either commercil concentrte (CON diet) or soyen curd residue silge (SCRS diet) t rtio of 80:20 on dry mtter sis. Ruminl fluid (50 ml) ws collected on dy 20 t 0 (efore feeding), 3 nd 6 hours fter the morning feeding vi stomch tue. The ph of the ruminl fluid ws immeditely mesured using ph meter (HM-10P, To Electronics Ltd., Jpn) fter collecting ech smple. The liquid frction of the ruminl fluid ws seprted y centrifuging t 8,000 x g for 10 minutes t 4⁰C. A 5 ml liquot of the liquid frction ws used to mesure the ruminl VFA content, nd 1 ml of the liquid frction ws mixed with 1 ml of 0.1 mol/l HCl for the 363 J. Anim. Sci. Adv., 2012, 2(4): ruminl mmoni N (NH 3 -N) nlysis. All of the smples were stored t -30⁰C until further nlysis. Mesurements of mino cid nd glucose kinetics On dy 21 of ech dietry tretment, primedcontinuous infusion [1-13 C]Leu nd [U- 13 C]glucose method ws conducted to determine the plsm Leu nd glucose kinetics simultneously over period of 4 hours, etween 3 nd 7 hours fter the morning feeding. Polyvinyl ctheters were plced in oth jugulr veins in the morning nd filled with

5 EFFECTS OF REPLACING CONCENTRATE WITH SOYBEAN CURD RESIDUE SILAGE ON RUMINAL sterile solution of 3.8% (w/v) trisodium citrte. A mixed sline solution of 7.2 μmol/kg BW 0.75 [1-13 C]Leu (L-leucine 1-13 C, 99 tom% excess 13 C; Cmridge Isotope Lortories, USA) nd 3.0 μmol/kg BW 0.75 [U- 13 C]glucose (D-glucose- 13 C 6, 99 tom% excess 13 C; Cmridge Isotope Lortories, USA) ws injected into the right jugulr ctheter s priming dose. This solution ws then continuously infused through the sme ctheter using multichnnel peristltic pump (AC-2120, Atto, Jpn) t rtes of 7.2 nd 3.0 μmol. kg BW h -1 for [1-13 C]Leu nd [U- 13 C]glucose, respectively. The infusion rte of the trcer solution ws recorded every 30 minutes throughout the infusion period. Blood smples (6 ml) were collected from the left jugulr ctheter immeditely efore the priming injection nd t 30-minutes intervl during the lst 2 hours of isotope infusion (5 to 7 hours fter feeding). The lood smples were plced in heprinized tues nd stored in ice. The plsm from the lood smple ws seprted y centrifugtion t 8,000 x g for 10 minutes t 4⁰C nd then stored t -30⁰C until further nlysis. Smple nlysis The chemicl components of the diets were nlyzed ccording to the Assocition of Officil Anlyticl Chemists (AOAC, 1990). The N contents in the diets, feces nd urine were nlyzed using the Kjeldhl method (Kjeltec 2100, Foss, Sweden). The fermenttion products of the silge were nlyzed using cold-wter extrcts (Ci et l., 1999). The ph vlues of the silge nd ruminl fluid were mesured using ph meter (HM-10P, To Electronics Ltd., Jpn). The lctic cid concentrtion of the silge ws mesured y the colorimetric method (Tylor, 1996), nd the sornce ws mesured using spectrophotometer (U-1000, Hitchi, Jpn) t 570 nm. The molr VFA concentrtions of the silge nd ruminl fluid were determined using gs chromtogrphy (HP-5890, Hewlett Pckrd, USA) fter stem distilltion. The voltile sic N (VBN) content of the silge ws determined sed on the stem distilltion method descried y Dhoudi et l. (2007). To ssess the qulity of silge, we clculted the Flieg point from the DM nd ph vlues (Yilmz et l., 2009) nd the V- score from the VBN/totl N nd VFA concentrtions (Tkhshi et l., 2005). The concentrtions of plsm Leu nd α- ketoisocproic cid (α-kic) nd enrichments of the plsm [1-13 C]Leu nd α-[1-13 C]KIC were determined y gs chromtogrphy mss spectrometry (QP-2010, Shimdzu, Jpn) with selected ion monitoring ccording to the procedures of Rocchiccioli et l. (1981) nd Clder & Smith (1988). The plsm [U- 13 C]glucose enrichment ws mesured using the methods of Tserng & Klhn (1983) with slight modifictions descried y Fujit et l. (2006). The plsm glucose concentrtion ws mesured y the glucose oxidse method (Huggett & Nixon, 1957). The plsm free AA nd ure concentrtions were determined using n utomted AA nlyzer (JLC- 500/V, JEOL, Jpn). The plsm non-esterified ftty cid (NEFA) concentrtions were determined enzymticlly using commercil dignostic kit (NEFA-C test, Wko, Jpn). Clcultions The turnover rtes of plsm Leu nd glucose (LeuTR nd GluTR) were clculted ccording to the eqution descried y Wolfe (1984): TR (mmol. kg BW h -1 ) = I x (1 / E 1) where I represents the infusion rtes of [1-13 C]Leu nd [U- 13 C]glucose nd E represents the isotope enrichments of the plsm α-[1-13 C]KIC, metolite of [1-13 C]Leu, nd plsm [U- 13 C]glucose t the stedy stte. Sttisticl Anlysis The men vlues nd stndrd errors of the men (SEM) were clculted for ll of the dt. All of the dt were nlyzed using the MIXED procedure in SAS (1996). The fixed effects in the model were the period, diet, nd the period x diet interction, nd the rndom effect ws the sheep. The results were considered significnt for P<0.05, nd the tendency ws defined s 0.05 P<0.10. Repeted sttements were used for the time course of the ruminl chrcteristics, nd the difference of the lest squre mens using Tukey s djustment ws used (P<0.05). 364 J. Anim. Sci. Adv., 2012, 2(4):

6 HARJANTI ET AL. Results Silge qulity nd N lnce The fermenttive chrcteristics of the SCRS re shown in Tle 3. The ph vlue ws 4.14, nd orgnic cids, such s lctic cid, cette, propionte nd utyrte, were detected. The effects of replcing commercil concentrte with SCRS on the N lnce re presented in Tle 4. Although oth diets were estimted to e isonitrogenous, the N intke determined t the end of the experiment tended (P=0.05) to e higher for the SCRS diet thn the CON diet. The fecl N excretion did not differ (P=0.17), ut the urinry N excretion tended to e higher (P=0.05) for sheep fed the SCRS diet thn those fed the CON diet. Both groups of sheep retined similr mounts of N (P=0.42), nd the N sorption nd digestiility did not differ (P=0.11 nd P=0.14, respectively) etween the two diets. Tle 3: Fermenttive chrcteristics of the soyen curd residue silge Item Amount Moisture (g/kg) 690 ph 4.14 Lctic cid (g/kg FM ) 40 Acette (g/kg FM) 1.1 Propionte (g/kg FM) 0.03 Butyrte (g/kg FM) VBN (g/kg totl N) 68 Flieg point 100 V-score 98 FM: fresh mtter VBN: voltile sic nitrogen Tle 4: Effects of feeding soyen curd residue silge on the nitrogen (N) lnce in sheep Items Tretment c SEM P vlue CON diet SCRS diet N intke (g. kg BW d -1 ) Fecl N (g. kg BW d -1 ) Urinry N (g. kg BW d -1 ) N sorption (g. kg BW d -1 ) N retention (g. kg BW d -1 ) N digestiility (%) Vlues represent the mens for n = 4. SEM: stndrd error of the men. c Tretment: mixed hy (orchrdgrss nd reed cnnrygrss) s the sl diet which is supplemented with either commercil concentrte (CON diet) or soyen curd residue silge (SCRS diet), t rtio of 80:20 on dry mtter sis. Ruminl chrcteristics The oserved chnges in the ruminl chrcteristics during the smpling period re provided in Figure 1. The effects of the two diets on the ruminl ph, NH 3 -N nd VFA concentrtions, represented s the men vlue of four sheep in three smpling periods (0, 3, nd 6 hours fter feeding), re shown in Tle 5. No 365 J. Anim. Sci. Adv., 2012, 2(4): interction ws oserved etween the dietry tretment nd the smpling time for the ruminl chrcteristics (P>0.10). The sheep tht consumed the SCRS tended (P=0.05) to hve higher ruminl ph thn those fed the CON diet. The ph vlues under oth tretments were ove 6.8 prior to feeding, nd the vlues dropped to pproximtely 6.7 t 3 hours fter feeding.

7 EFFECTS OF REPLACING CONCENTRATE WITH SOYBEAN CURD RESIDUE SILAGE ON RUMINAL P vlue Tle 5: Effects of feeding soyen curd residue silge on the ruminl chrcteristics of sheep Tretment c Item CON diet SCRS diet SEM Tretment Time Tretment x Time ph NH 3 -N (mmol/l) Totl VFA (mmol/l) Individul VFA concentrtions (mmol/l) Acette Propionte < Isoutyrte Butyrte Isovlerte Vlerte Acette:propionte rtio Vlues represent the mens for n = 4. SEM: stndrd error of the men. c Tretment: mixed hy (orchrdgrss nd reed cnnrygrss) s the sl diet which is supplemented with either commercil concentrte (CON diet) or soyen curd residue silge (SCRS diet), t rtio of 80:20 on dry mtter sis. NH 3 -N: mmoni nitrogen; VFA: voltile ftty cid 366 J. Anim. Sci. Adv., 2012, 2(4):

8 HARJANTI ET AL. Comprle NH 3 -N concentrtions were found for oth groups (P=0.22); the NH 3 -N concentrtions were incresed t 3 hours fter feeding nd eventully returned to the previous vlues under oth dietry tretments. The totl VFA concentrtion in the rumen did not differ significntly (P=0.13) etween the two diets. The cette concentrtions were similr for the two diets, ut the propionte concentrtion ws higher (P=0.02) in the sheep fed the SCRS thn the CON diet, resulting in lower cette:propionte rtio (P=0.03) for the SCRS thn CON diet. The utyrte concentrtion ws lower (P=0.02) for the SCRS diet compred to the CON diet, wheres the vlerte concentrtion ws higher (P=0.02) nd other rnched-chin VFA (isoutyrte nd isovlerte) concentrtions tended to e higher (P=0.05 nd P=0.06, respectively) under the SCRS tretment. The totl VFA concentrtion nd the concentrtions of cette, propionte, utyrte, nd vlerte were incresed t 3 hours fter feeding nd eventully returned towrd previous vlues. The isoutyrte nd isovlerte concentrtions were grdully decresed t 6 hours fter feeding. Pre-isotope infusion plsm concentrtion of lood metolites The plsm AA, ure nd NEFA concentrtions determined t the pre-isotope infusion period t 3 hours fter feeding re presented in Tle 6. Tle 6: Effects of feeding soyen curd residue silge on the plsm mino cid, ure nd non-esterified ftty cid concentrtions during pre-isotope infusion in sheep Tretment c Item CON diet SCRS diet SEM P vlue Essentil AA (μmol/l) Threonine Vline Methionine Isoleucine Leucine Phenyllnine Histidine Lysine Non-essentil AA (μmol/l) Serine Asprgine Glutmic cid Glutmine Glycine Alnine Tyrosine Arginine Proline Ure, mmol/l NEFA, meq/l Vlues represent the mens for n = 4. SEM: stndrd error of the men. c Tretment: mixed hy (orchrdgrss nd reed cnnrygrss) s the sl diet which is supplemented with either commercil concentrte (CON diet) or soyen curd residue silge (SCRS diet), t rtio of 80:20 on dry mtter sis. AA: mino cid NEFA: non-esterified ftty cid The plsm isoleucine, Leu, phenyllnine, nd tyrosine concentrtions were lower (P<0.05) 367 J. Anim. Sci. Adv., 2012, 2(4): nd the plsm methionine, histidine, rginine, nd proline concentrtions tended to e lower (P<0.10)

9 EFFECTS OF REPLACING CONCENTRATE WITH SOYBEAN CURD RESIDUE SILAGE ON RUMINAL in the sheep fed the SCRS thn CON diet; the concentrtions of other AA were comprle etween the diets. The plsm ure concentrtion ws higher (P=0.02) in the sheep fed the SCRS thn CON diet, nd the plsm NEFA concentrtion did not differ (P=0.38) etween the diets. Amino cid nd glucose kinetics The plsm Leu nd α-kic concentrtions nd plsm α-[1-13 C]KIC enrichment were in stedy stte during the lst 2 hours of isotope infusion (Figure 2). The plsm Leu nd α-kic concentrtions did not differ (P=0.18 nd P=0.13, respectively) etween the diets (Tle 7). The sheep fed the SCRS hd similr (P=0.74) plsm LeuTR with those of sheep fed the CON diet. The plsm glucose concentrtion nd [U- 13 C]glucose enrichment were essentilly constnt during the lst 2 hours of the isotope infusion (Figure 3). The plsm glucose concentrtion nd GluTR did not differ (P=0.68 nd P=0.27, respectively) etween the two diets. Tle 7: Effects of feeding soyen curd residue silge on the plsm leucine (Leu) nd glucose kinetics in sheep Tretment c Item CON diet SCRS diet SEM P vlue Leu concentrtion (μmol/l) α-kic concentrtion (μmol/l) LeuTR (mmol. kg BW h -1 ) Glucose concentrtion (mmol/l) GluTR (mmol. kg BW h -1 ) Vlues represent the mens for n = 4. SEM: stndrd error of the men. c Tretment: mixed hy (orchrdgrss nd reed cnnrygrss) s the sl diet which is supplemented with either commercil concentrte (CON diet) or soyen curd residue silge (SCRS diet) t rtio of 80:20 on dry mtter sis. α-kic: α-ketoisocproic cid LeuTR: leucine turnover rte GluTR: glucose turnover rte Discussion Soyen curd residue silge qulity The SCRS ws well preserved, s indicted y the low ph, high lctic cid content, high Flieg point nd V-score. The lctic cid concentrtion in our SCRS ws similr to tht found y Amh et l. (1996), ut it ws higher thn tht found y Xu et l. (2001). During the fermenttion process, some hemicellulose nd cellulose were roken down into vrious simple sugrs nd smller end products, such s lctic cid, cette nd utyrte (Co et l., 2009). The NDF content in our SCRS (150 g/kg DM) ws comprle to the NDF content reported from rw SCR (145 g/kg DM) y O Toole (1999). Nitrogen lnce Previous studies (Oltjen & Putnm, 1966; Clifford & Tillmn, 1968; Prior, 1976; Knus et l., 2002) hve shown tht inclusion soy protein in the diet of ruminnts enhnces the N intke, urinry N excretion, N retention nd N sorption. Sno et l. (2009) studied the effect of different N sources supplementtion (ure nd soyen mel) in the isonitrogenous design nd found lrge urinry N excretion difference etween tretments, indicting chnge in N retention. In the present study, replcing the commercil concentrte with SCRS did not ffect the mount of N sored nd retined, wheres it did slightly ffect the urinry N excretion. Hence, our findings demonstrte the fesiility of replcing commercil concentrte with soyen curd residue silge to mintin positive N lnce. Ruminl chrcteristics The ruminl ph vlues of oth groups of sheep were ove 6.5 for ll of the smpling times, nd this is the optiml ph for norml ruminl fermenttion. 368 J. Anim. Sci. Adv., 2012, 2(4):

10 HARJANTI ET AL. c Time fter feeding (hour) Time fter feeding (hour) Fig. 1: Time-course chnges of ruminl chrcteristics in sheep fed the SCRS diet ( ) nd the CON diet ( ). SCRS diet, mixed hy supplemented with soyen curd residue silge; CON diet, mixed hy supplemented with commercil concentrte. A rtio of 80:20 (dry mtter sis) ws used for oth diets. Ruminl fluid smples were collected t 0 (efore feeding), 3 nd 6 hours fter morning feeding vi stomch tue. Vlues re expressed s men ± SEM for n = 4. Different letters (,, c) indicte significnt different (P<0.05) etween time fter feeding within ech tretment. 369 J. Anim. Sci. Adv., 2012, 2(4):

11 EFFECTS OF REPLACING CONCENTRATE WITH SOYBEAN CURD RESIDUE SILAGE ON RUMINAL Thus, replcing the commercil concentrte with the SCRS did not impir ruminl fermenttion; similr response oserved y Xu et l. (2001). The SCRS nd CON diets in the present study were pproximtely isonitrogenous nd isoenergetic; thus, similr N compounds should e ville for NH 3 -N production in the rumen. This finding is consistent with the results of Iphrrguerre et l. (2005) who reported tht the concentrtion of NH 3 -N in the rumen fluid only ffected y the level of dietry CP intkes. Time fter strting [U- 13 C] glucose infusion (hour) Fig. 2: Time course chnges of plsm leucine (Leu) nd α-ketoisocproic cid (α-kic) concentrtions, plsm [1-13 C]Leu nd α-[1-13 C]KIC enrichments during the lst 2 hours of isotope infusion in sheep fed the SCRS diet ( ) nd the CON diet ( ). SCRS diet, mixed hy supplemented with soyen curd residue silge; CON diet, mixed hy supplemented with commercil concentrte. A rtio of 80:20 (dry mtter sis) ws used for oth diets. Vlues re expressed s men ± SEM for n = 4. Time fter strting [1-13 C]Leu infusion (hour) Fig. 2: Time course chnges of plsm leucine (Leu) nd α-ketoisocproic cid (α-kic) concentrtions, plsm [1-13 C]Leu nd α-[1-13 C]KIC enrichments during the lst 2 hours of isotope infusion in sheep fed the SCRS diet ( ) nd the CON diet ( ). SCRS diet, mixed hy supplemented with soyen curd residue silge; CON diet, mixed hy supplemented with commercil concentrte. A rtio of 80:20 (dry mtter sis) ws used for oth diets. Vlues re expressed s men ± SEM for n = J. Anim. Sci. Adv., 2012, 2(4):

12 HARJANTI ET AL. Voltile ftty cids re produced in the rumen s the end product of microil fermenttion, nd their production lrgely depends on the type of crohydrte ingested (Djikstr, 1994). The lrge increment in the ruminl propionte concentrtion in the sheep fed the SCRS diet could e ttriuted to the highly solule crohydrte nd strch contents (Yng, 2005). Although the NDF concentrtion in ruminnt diet is positively correlted with the mount of VFA produced in the rumen (Beuchemin, 1991), the lower NDF content for the SCRS (150 g/kg DM) compred to the commercil concentrte (207 g/kg DM) did not lter the totl VFA concentrtion in the rumen. Replcing the commercil concentrte with the SCRS did not significntly influence the ruminl cette concentrtion, indicting similr efficiencies of energy-source production for oth diets. Pre-isotope infusion plsm concentrtion of lood metolites The primry source of dietry protein (Schelling et l., 1967) ffects the plsm AA concentrtions in sheep. For the different protein sources in the present study, the lower concentrtions of certin plsm AAs in the sheep fed the SCRS diet might not e indictive of the totl AA supply ecuse the protein supply in oth tretments ws similr nd sufficient. Thus, the oserved lower AA levels were most likely due to the limiting AAs in the diet (Lpiere & Loley, 2001), which, in soyen-sed diets, re sulfurcontining AAs (methionine nd cysteine) (M et l., 1996). Moreover, Lpierre nd Loley (2001) suggested tht the concentrtion of plsm AA fter feeding vried depend on the rte of digestion nd AA ctolism of the dietry protein source. Furthermore, these uthors noted tht pproximtely 46% of AA from highly degrdle type of protein such s silge did not pper in the peripherl vein, wheres for concentrte the mount vried etween 23% nd 24%. Although the sheep fed the SCRS diet in our study exhiited lower concentrtions of certin plsm AAs, replcing the commercil concentrte with the SCRS did not induce n AA imlnce, sed on the positive N lnce nd similr N retention for oth of the diets. The use of different protein sources ffected the plsm ure concentrtion, which is metolic end product of protein ctolism or the product of solule protein fermenttion in the rumen. Although the ruminl NH 3 -N concentrtions within the smpling periods (0, 3, nd 6 hours fter feeding) did not differ significntly etween the two diets, the high plsm ure concentrtion for the SCRS diet, reflected y the rpid degrdtion of N compounds nd the rpid sorption of NH 3 -N through the rumen, resulted in n incresed level of plsm ure. Furthermore, the N excretion vi the urine tended to increse over time. Similrly, Leiholz nd Cook (1967) showed tht elevted plsm ure concentrtions incresed the N excretion rte vi the urine nd the rte of return of N to the rumen s ure in the sliv in sheep fed lflf hy nd concentrte. The circulting plsm NEFA concentrtion ws mrkedly relted to the nutritionl sttus. The plsm NEFA concentrtion increses with high rtes of lipolysis, such s occurs with fsting, negtive energy lnce or stress (Trenkle & Kuhlemeier, 1966). Hence, the comprle plsm NEFA concentrtions indicted comprle nutritionl sttus etween sheep fed the SCRS nd CON diet. Amino cid nd glucose kinetics In the present study, oth plsm [1-13 C]Leu nd α-[1-13 C]KIC enrichments were determined, ut only plsm α-[1-13 C]KIC enrichment is discussed ecuse it represents n ccurte index of the intr- nd extr-cellulr mino cid exchnge (Mgni et l., 1994). The plsm LeuTR oserved in our study were comprle to previous studies using single-isotope infusion of [1-13 C]Leu to determine protein kinetics in sheep (Sno et l., 2004; 2009); similrly, the plsm GluTR oserved in our study were comprle to those from previous study on glucose kinetic in sheep determined using single-isotope infusion of [U- 13 C] glucose (Sno et l., 2000). These results indicte tht the doule isotope used in our experiment did not lter either the plsm Leu or 371 J. Anim. Sci. Adv., 2012, 2(4):

13 EFFECTS OF REPLACING CONCENTRATE WITH SOYBEAN CURD RESIDUE SILAGE ON RUMINAL glucose kinetics of the sheep. The use of different protein sources resulted in vrying plsm AA concentrtions, which could influence the turnover rte of proteins (Nielsen et l. 1993). Although reports on the AA kinetics of soyen-sed diets in ruminnts re limited, the results from other studies in pigs (Deutz et l., 1998) nd humns (Bos et l., 2003) showed tht the AAs from soyen-sed diets re rpidly digested nd sored, indicting n enhnced AA ctolism. The low concentrtion of plsm AAs in the sheep fed the SCRS diet could hve een cused y the utiliztion of AAs s n energy sustrte to support protein turnover within the tissues; thus, the plsm LeuTR mesured were comprle etween the two diets. In ddition, the dietry tretments were pproximtely isoenergetic nd isonitrogenous, which could hve contriuted to the comprle plsm LeuTR for the two diets ecuse the plsm LeuTR ws positively correlted with the dietry protein nd metolizle energy intke in sheep (Liu et l., 1995) nd cows (Lpierre et l., 2002). Similrly, Sno et l. (2009) reported comprle plsm LeuTR for different protein sources (ure nd soyen mel) in sheep fed the isoenergetic nd isonitrogenous diets. The GluTR is influenced y severl fctors, including the energy intke nd supply of gluconeogenetic sustrte to the liver (Ortiques- Mrty et l., 2003). The propionte produced in the rumen is mjor glucogenic sustrte nd precursor of de novo glucose synthesis (Herein et l., 1978). In the present study, lthough the glucogenic propionte concentrtion in the rumen ws higher for the SCRS thn CON diet, the plsm GluTR were similr for oth of the diets. In ccordnce with this result, n incresing supply of glucogenic propionte did not systemticlly increse plsm GluTR in steers (Sel & Prker, 1994) nd sheep (Linington et l., 1998). Herein et l. (1978) nd Sel et l. (1992) pointed out tht similr energy intke resulted in similr plsm GluTR in sheep nd growing steers. Thus, the isoenergetic diets in the present study did not significntly influence the plsm GluTR. The comprle results for the plsm Leu nd glucose kinetics indicted similr nutritive vlue for the two diets. Similr plsm Leu nd glucose kinetic responses could e chieved y replcing commercil concentrte with SCRS in the sme mount of metolizle energy intke. In conclusion, replcing commercil concentrte with SCRS in the diet of sheep did not impir ruminl fermenttion, nd enhnced the glucogenic propionte concentrtion. The inclusion of SCRS in the diets of sheep could ensure positive N lnce nd resulted in similr plsm AA nd glucose kinetics with commercil concentrte when formulted in the sme energy intke. Thus, feed cost cn e reduced y replcing commercil concentrte with soyen curd residue silge. Furthermore, technologicl developments in the use of residue nd y-products will help to improve the feed self-sufficiency rtes nd mke niml griculture more sustinle. Acknowledgements The uthors re grteful to Hirkw Food Co, Ltd. for providing the soyen curd residue for the experiments. References AOAC. 1990: Officil Methods of Anlysis.15th ed. Assoc. Off. Anl. Chem., Arlington, VA. Amh K, Sski Y, Segw T (1996). Utiliztion of tofu (soyen curd) y-product s feed for cttle. Food nd Fertilizer Tech. Accessed My 27, Beuchemin KA (1991). Effects of dietry neutrl detergent fier concentrtion nd lff hy qulity on chewing, rumen function, nd milk production of diry cows. J. Diry Sci., 74: Bequette BJ, Metclf JA, Wry-Chen D, Bckwell FRC, Sutton JD, Lomx MA, McRe JC, Loley GE (1996). Leucine nd protein metolism in the lctting diry cow mmmry glnd: responses to supplementl dietry crude protein intke. J. Diry Res., 63: Bergen WG, Hennemn HA, Mgee WT (1973). Effect of dietry protein level nd protein source on plsm nd tissue free mino cids in growing sheep. J. Nutr., 103: Bos C, Metges CC, Gudichon C, Petzke KJ, Pueyo ME, Morens, C, Everwnd J, Benmouzig R, Tome D (2003). Postprndil kinetics of dietry mino cids re the min determinnt of their metolism fter soy or milk protein ingestion in humns. J. Nutr., 133: Ci Y, Benno Y, Ogw M, Kumi S (1999). Effect of pplying lctic cid cteri isolted from forge crops on fermenttion chrcteristics nd eroic deteriortion 372 J. Anim. Sci. Adv., 2012, 2(4):

14 HARJANTI ET AL. of silge. J. Diry Sci., 82: Clder AG, Smith A (1988). Stle isotope rtio nlysis of leucine nd ketoisocproic cid in lood plsm y gs chromtogrphy/mss spectrometry. Use of tertiry utyldimenthylsilyl derivtives. Rpid Comm. Mss Spectrom., 2: Co Y, Tkhshi T, Horiguchi K (2009). Effects of ddition of food y-products on the fermenttion qulity of totl mixed rtion with whole crop rice nd its digestiility, preference, nd rumen fermenttion in sheep. Anim. Feed Sci. Technol., 151: Cho JH, Min BJ, Chen YJ, Yoo JS, Wng Q, Kim JD, Kim IH (2007). Evlution of FSP (fermented soy protein) to replce soyen mel in wened pigs: growth performnce, lood ure nitrogen nd totl protein concentrtions in serum nd nutrient digestiility. Asin-Aust. J. Anim. Sci., 20: Clifford AJ, Tillmn AD (1968). Ure nd isolted soyen protein in sheep purified diets. J. Anim. Sci., 27: Deutz NEP, Bruins MJ, Soeters PB (1998). Infusion of soy nd csein protein mels ffects interorgn mino cid metolism nd ure kinetics differentilly in pigs. J. Nutr., 128: Dhoudi A, Monser L, Sdok S, Adhoum N (2007). Vlidtion of flow-injection-gs diffusion method for totl voltile sic nitrogen determintion in sefood products. J. Food Chem., 103: Djikstr J (1994). Production nd sorption of voltile ftty cids in the rumen. Livest. Prod. Sci., 39: Fujit T, Kjit M, Sno H (2006). Responses of whole ody protein synthesis, nitrogen retention nd glucose kinetics to supplementl strch in gots. Comp. Biochem. Physiol., 144B: Given DI, Rulquin H (2004) Utilistion y ruminnts of nitrogen compounds in silge-sed diets. Anim. Feed Sci. Technol., 114: Hrris PM, Skene PA, Buchn V, Milne E, Clder AG, Anderson SE, Cornell A, Loley GE (1992). Effect of food intke on hind-lim nd whole ody protein metolism in young growing sheep: chronic studies sed on rterio-venous techniques. Br. J. Nutr., 68: Herein JM, Vn Mren RW, McGillird AD, Young JW (1978). Rumen propionte nd lood glucose kinetics in growing cttle fed isoenergetic diets. J. Nutr., 108: Huggett AG, Nixon DA (1957). Enzymtic determintion of lood glucose. J. Biochem., 66: 12. Iphrrguerre IR, Clrk JH, Freemn DE (2005). Vrying protein nd strch in the diet of diry cows. 1. Effects on ruminl fermenttion nd intestinl supply of nutrients. J. Diry Sci., 88: Knus WF, Beermnn DH, Tedeschi LO, Czjkowski M, Fox DG, Russel JB (2002). Effects of ure, isolted soyen protein nd lood mel on growing steers fed cornsed diet. Anim. Feed Sci. Techol., 102: Lpierre H, Loley GE (2001). Nitrogen recycling in the ruminnt: A Review. J. Diry Sci., 84: E223-E236. Lpierre H, Blouin JP, Bernier JF, Reynolds CK, Dureuil P, Loley GE (2002). Effect of supply of metolizle protein on whole ody nd splnchnic leucine metolism in lctting cows. J. Diry Sci., 85: Leiholz J, Cook CF (1967). Free mino cids, mmoni nd ure concentrtions in the lood plsm of strved lms. J. Nutr., 93: Linington MJ, Meyer JHF, vn der Wlt JG (1998). Ruminl VFA production rtes, whole ody metolite kinetics nd lood hormone concentrtions in sheep fed high nd low fire diets. S. Afr. J. Anim. Sci., 28: Liu SM, Loley GE, McLeod NA, Kyle DJ, Chen XB, Ørskov ER (1995). Effects of long-term protein excess or deficiency on whole ody protein turnover in sheep nourished y intrgstric infusion of nutrients. Br. J. Nutr., 73: M CY, Liu WS, Kwok KC, Kwok F (1997). Isoltion nd chrcteriztion of protein from soymilk residue (okr). Food Res. Int., 29: MAFF. Ministry of Agriculture, Forestry nd Fisheries Reserch Council (1995). Stndrd tles of feed composition in Jpn. Centrl ssocition of livestock industry. Jpn. Mgni F, Arnoldi L, Glti G, Kienle M (1994). Simultneous determintion of plsm levels of α- ketoisocproic cid nd leucine nd evlution of α-[1-13 C]ketoisocproic cid nd [1-13 C]leucine enrichment y gs chromtogrphy-mss-spectometry. Anl. Biochem., 220: Mly PS, Perek AE, Weisjerg MR, Hvelplund T, Mdsen J (2003). Digestion nd pssge kinetics of fire in mture diry heifers mintined on poor qulity hy s ffected y the source nd level of nitrogen supplementtion. Anim. Feed Sci. Technol., 109: Nielsen K, Kondrup J, Elsner P, Juul A, Jensen ES (1994). Csein nd soyen protein hve different effects on whole ody protein turnover t the sme nitrogen lnce. Br. J. Nutr., 72: NRC (1985). Nutrient Requirements of Sheep, 6 th revised edition. Ntionl Acdemy Press, Wshington, DC. Ortigues-Mrty I, Vernet J, Mjdou L (2003). Whole ody glucose turnover in growing nd non-productive dults ruminnts: met-nlysis nd review. Reprod. Nutr. Dev., 43: Oltjen RR, Putnm PA (1966). Plsm mino cid nd nitrogen retention y steers fed purified diets contining ure or isolted soy protein. J. Nutr., 89: O Toole D (1999). Review: Chrcteristics nd use of okr, the soyen residue from soy milk production. J. Agric. Food Chem., 47: Prior RL (1976). Effects of dietry soy or ure nitrogen nd feeding frequency on nitrogen metolism, glucose metolism nd urinry metolite excretion in sheep. J. Anim. Sci., 42: J. Anim. Sci. Adv., 2012, 2(4):

15 EFFECTS OF REPLACING CONCENTRATE WITH SOYBEAN CURD RESIDUE SILAGE ON RUMINAL Rocchiccioli F, Leroux JP, Crtier P (1981). Quntittion of 2-ketocids in iologicl fluids y gs chromtogrphy chemicl ioniztion mss spectrometry of o- trimethylsilyl-quinoxlinol derivtives. Biomed. Mss Spectrom., 8: Sno H, Konno S, Shig A (2000). Chromium supplementtion does not influence glucose metolism or insulin ction in response to cold exposure in mture sheep. J. Anim. Sci., 78: Sno H, Kjit M, Fujit T (2004). Effect of dietry protein intke on plsm leucine flux, protein synthesis, nd degrdtion in sheep. Comp. Biochem. Physiol., 139B: Sno H, Shiski S, Swd H (2009). The effect of the source of nitrogen supplementtion on nitrogen lnce, rtes of plsm leucine turnover, protein synthesis nd degrdtion in sheep. Arch. Anim. Nutr., 63: SAS SAS/STAT softwre: chnges nd enhncement through Relese SAS Institute Inc. Cry. NC, USA. Schelling GT, Hinds FC, Htfield EE (1967). Effect of dietry protein levels, mino cid supplementtion nd nitrogen source upon the plsm free mino cid concentrtions in growing lms. J. Nutr., 92: Sel CJ, Prker DS, Avery PJ (1992). The effect of forge nd forge-concentrte diets on rumen fermenttion nd metolism of nutrients y mesenteric nd portldrined viscer in growing steers. Br. J. Nutr., 67: Sel CJ, Prker DS (1994). Effect of intrruminl propionic infusion on metolism of mesenteric nd portl drined-viscer in growing steers fed forge diet: I. Voltile ftty cids, glucose nd lctte. J. Anim. Sci., 72: Tkhshi T, Horiguchi K, Goto M (2005). Effect of crushing unhulled rice nd the ddition of fermented juice epiphytic lctic cteri on the fermenttion qulity of whole crop rice silge, nd its digestiility nd rumen fermenttion sttus in sheep. Anim. Sci. J., 76: Tylor KACC (1996). A simple colorimetric ssy for murmic cid nd lctic cid. Appl. Biochem. Biotechol., 56: Trenkle A, Kuhlemeier KV (1996). Reltionship of rumen voltile cids, lood glucose, nd plsm nonesterified ftty cids in sheep. J. Anim. Sci., 25: Tserng KY, Klhn SC (1983). Estimtion of glucose cron recycling nd glucose turnover with [U- 13 C]glucose. Am. J. Physiol., 245: E476-E482. Wolfe RR (1984). Trcers in Metolic Reserch. Rdioisotope nd stle isotope/mss spectrometry methods. Aln R. Liss, Inc, New York. Xu CC, Suzuki H, Toyokw K (2001). Chrcteristics of ruminl fermenttion of wethers fed tofu cke silge with ethnol. Anim. Sci. J., 72: Yng CMJ (2005). Soyen milk residue ensiled with penut hulls: fermenttion cids, cell wll composition, nd silge utiliztion y mixed ruminl microorgnisms. Biores. Technol., 96: Yilmz A, Altinok S, Kocs Z (2009). An investigtion on qulity prmeters of the silges mde y corn nd soyen grown in different seeding rtes. J. Anim. Vet. Adv., 8: J. Anim. Sci. Adv., 2012, 2(4):