INFLUENCE OF DAILY SUPPLEMENTATION OF CARIB PODS (Ceratonia Siliqua L.) AS POLYPHENOL RICH PLANTS ON RUMEN FERMENTATION AND LAMBS GROWTH PERFORMANCE

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1 Scientific Ppers Series Mngement, Economic Engineering in Agriculture nd Rurl Development INFLUENCE OF DAILY SUPPLEMENTATION OF CARIB PODS (Certoni Siliqu L.) AS POLYPHENOL RICH PLANTS ON RUMEN FERMENTATION AND LAMBS GROWTH PERFORMANCE Mohmed Smir KHALEL 1, Mohmed Helmy YACOUNT 1, Aymn Adel-Mohsen HASSAN 1, Amr Mohmed SHWERAB 1, Dorin MOCUTA 2, Adrin TUREK-RAHOVEANU 2 1 Animl Production Reserch Institute, Agriculture Reserch Center, Deprtment of y-products Utiliztion, Dokki, Giz, Egypt, Emils: drmohmedes@yhoo.com; helmy_ycout555@hotmil.com; ymnn19@hotmil.com; mr.shwer@yhoo.com 2 University of Agriculturl Sciences nd Veterinry Medicine, Fculty of Mngement nd Economic Engineering, Buchrest, 59 Mrsti Boulevrd, District 1, , Romni, Emil:dorinmocut@yhoo.com Corresponding uthor: ymnn19@hotmil.com Astrct This study ws conducted to evlute the influence of dily supplementtion of cro Pods (Certoni Siliqu L.) t the rte of 0, 25, 50 nd 100 gm on digestiility coefficients, rumen fermenttion, degrdtion kinetics, microil protein synthesis, some lood prmeters, ntioxidnt ctivity nd lms growth performnces. Three rms were consigned for the digestiility trils, while other three femles with rumen fistul were used for the in situ trils. Fourty lms (6-8 months) were used for feeding trils (90 dys). Dt showed tht dily supplement with 50gm cro pods hd higher digestion coefficients, lower rumen NH 3-N, ut higher TVFA's concentrtion, nd microil- N synthesis compred with other supplementtion rts. In ddition to more effective degrdility of DM nd CP, feed nd economic efficiencies nd dily weight gin. Supplementtion with 100 g cro pods ws the worst one. Insignificnt differences were found for lmost lood constitutes mong the experimentl diets, they were in the norml rnges. So, it could e concluded to dily supplement sheep with 50gm of cro pods in order to chieve good productive performnce of lms. However, it still needs to crry long term trils on the field of met or milk production. Key words: cro pods, digestiility, rumen fermenttion, degrdtion kinetics growing lms, lood constitutes INTRODUCTION Cro tree (Certoni siliqu L.) is ntive to the Mediterrnen re, the min producers of cro fruits re Spin, Itly, Portugl, Morocco nd Greece [23]. Trditionlly, cros were cultivted for humn nd niml nutrition, while nowdys cro seeds nd pods hve wide ppliction s nturl food dditives nd stilizer gents in the food industry, e.g. coco sustitute, gums, sugrs, everges or phrmceuticl nd cosmetic industries [55]. However, the presence of tnnins in cro pods with their properties such s ntidirrhel, nticteril, ntioxidnt nd free-rdicl scvenging nd ntiprolifertive ctivity in liver cells my hve some eneficil effects on humn nd niml helth [12] [16]. Cro pods hve een wide used in niml nutrition, for sheep [33], lms [47], rits [26], nd poultry [44]. Tnnins, cn e divided ccording to their chemicl structure into four mjor groups: condensed tnnins, hydrolysle tnnins, phlorotnnins nd complex tnnins [50]. They hd the ility to form insolule complexes with proteins or digestive enzymes which reduce digestiility of dietry proteins [31]. On the other hnd, dietry tnnins pper to hve some other eneficil effects in ruminnts, they minly prevent excessive ruminl degrdtion of dietry proteins, reduce the ctivities of microil proteses [8].The present pper imed to evlute the effect of four supplementtion rtes (0, 25,

2 Scientific Ppers Series Mngement, Economic Engineering in Agriculture nd Rurl Development nd 100 gm.) of cro pods on digestiility, rumen fermenttion, degrdtion kinetics, some lood constitutes nd lms performnce. MATERIALS AND METHODS A totl of 40 mle growing lms (8 months) with n verge weight of 24.56±0.13 kg were rndomly llotted into 4 groups (10 nimls ech) to study their growth performnce. Four experimentl concentrte feed mixtures (CFM) were formulted with 14 % CP. Lms were fed the CFM's nd rice strw (RS) nd dily supplemented with 0 or 25, 50 or 100 mg cro pods for 90 dys ccording to [42]. A digestiility tril ws conducted y 12 dult rms (3 rms for ech group), they were individully llotted in metolic cges for 3 weeks s dpttion period followed y 7 dys for collecting urine nd feces. Animls were fed s ove t 8.00 m nd 4.00 pm., while wter ws ville ll over the dy. Tle 1 illustrted composition of CFM nd its chemicl composition nd cro pods. Feces nd urine were collected quntittively once dy efore the morning mel t 8.00 m nd weight fresh. Dily smples representtive 20 % of fresh feces nd urine, then the seven dys comined collection of smples were stored t - 20 C nd kept for routine nlysis. Feeds nd fecl smples were nlyzed for crude protein (CP) y Kjeldhl, crude fier (CF), ether extrct (EE) nd sh, while urine ws nlyzed for nitrogen (N) ccording to AOAC [4]. Cell wll (neutrl detergent fier (NDF), cid detergent fier (ADF), cid detergent lignin (ADL), cellulose nd hemicelluloses content ws determined y [58] procedure. Vlues of the totl digestile nutrients (TDN) were clculted ccording to the clssic formul of Cheeke on dry mtter sis [15]. Live ody weight (g) nd dily feed consumption (g) were individully recorded. Feed conversion ws clculted s feed consumption (g) / weight gin (g). Economicl efficiency (Y) ws clculted ccording to the following eqution: Y = (A- B) / B x 100, where (A) is the selling cost of 274 the otined gin nd (B) is the feeding cost of this gin. Tle 1. Composition of CFM & its chemicl nlysis nd cro pods (on DM sis). Ingredients Cro pods CFM (g/kg) Yellow corn 330 Brley grin 200 Whet rn 170 Soyen mel (44 % CP) 140 Olive cke 70 Molsses 50 Limestone 20 Sodium chloride 15 Minerl-vitmin premix 2 5 Chemicl nlysis (g/kg DM): Dry mtter Orgnic mtter Crude protein Crude fier Ether extrct NFE Ash NDF ADF ADL Hemicellulose Cellulose Neutrl 68.4 detergent solule Totl phenolic Totl tnnins One kilogrm of premix contin: Vit. A IU, Vit. D IU, Vit. E 1000mg, Vit. B mg B mg, Vit. B6 100 mg, Vit. B12 10 mg, Pntothenic cid 3.33 g, Biotin 3mg, Folic cid 0.83 g, Zn g, Mn 5 g, Fe 12.5 g, Cu 0.5 g, Se 16.6 mg nd Mg 66.7 g. 1 NFE = 100 (% CP+ % EE + % Ash + % NDF) Rumen fermenttion trils Three fistulted Brki femle sheep (45.5±0.5kg BW) were used for rumen fermenttion trils. Smples of rumen liquor were tken t 0, 1, 3 nd 6 h post feeding from for ech tretment, to e nlyzed for ph using Orion 680 digitl ph meter. Rumen fluid smples were preserved for mmoni nitrogen (NH3-N) determintion ccording to Preston [46], totl voltile ftty cid

3 Scientific Ppers Series Mngement, Economic Engineering in Agriculture nd Rurl Development (TVFA s) y using stem methods [61]. Microil counts (cteri nd protozo) in the ruminl fluid were determined using counting cell (Hwskley, UK) s descried y Demeyer [17]. Rumen volume ws determined y the colorimetric method using Cr-EDTA ccording to El-Shzly [22]. Synthesized microil protein (MP g/dy) in the rumen ws clculted ccording to Borhmi [13]. Nylon gs technique ws used to determine degrdility kinetics of DM nd CP. Two polyester gs (7x15 cm) with pore size of 45 µm were used for ech incution time. Approximtely 5 g of ir-dried diet (ground to 2 mm) were plced in ech g. All gs were incuted in the rumen, then they were withdrwn fter 3, 6, 12, 24, 48 nd 72 h; rinsed in tp wter until the wter ecme cler, then they were squeezed gently. Microorgnisms ttched to the residul smple were eliminted y freezing t -20ºC [32]. Zero-time wshing losses () were determined y wshing 2 gs in running wter for 15 min. The degrdtion kinetics of DM nd CP were estimted (in ech g) y fitting the disppernce vlues to the eqution p= + (1-e) s proposed y McDonld [35]. The effective degrdility (ED) for tested rtions were estimted from the eqution of McDonld [35], ED = + c/ (c + k), where ED (effective degrdility), (rpidly degrded frction), (slowly degrded frction), c (rte of degrdtion) nd k is the out flow rte ssumed to e 0.05/h for concentrte under the feeding condition in this study. Blood smples were collected t the end of the experimentl period from ll lms. Blood smples were otined from the jugulr vein of the lms in the morning efore ccess to feed nd wter. Serum ws otined y centrifugtion of lood nd stored t 20 C until the nlysis time. Glucose concentrtion ws determined ccording to Trinder [54]; serum cholesterol [53]; serum totl protein (TP) ccording to Henry [29], lumin (A) concentrtion ccording to Doums [20]. Kidney function ws evluted y mesuring lood ure using the colorimetric methods of Henry nd Todd [30]. Liver function ws ssessed y mesuring the ctivities of sprtte minotrnsferse (AST) nd lnine mino trnsferse (ALT) y the method of Reitmn nd Frnkel [48]. Antioxidnt enzyme ctivities The concentrtion of SOD ws determined s descried y Ogunugfor [43]. The ctlse ctivity (CAT) ws determined s descried y Usoh et l., (2005) [57]. The ctivity of glutthione peroxidse (GPx) nd reduced glutthione were determined y the method of Beutler [10]. Lipid peroxide concentrtion mesured s thiorituric cid rective sustnces (TBARS) ccording to Trott [56]. Sttisticl Anlysis The otined dt were sujected to sttisticl nlysis using generl liner models (GLM) procedure of SAS [49]. Significnt differences mong mens were seprted using LSD test ccording to Duncn [21] nd significnce ws declred t P<0.05. RESULTS AND DISCUSSIONS Digestiility coefficients All vlues of digestiility coefficient were significntly incresed (P<0.05) for sheep supplemented with 50 gm cro pods compred to other groups (Tle 2). Insignificnt differences were found etween the 25 nd 50gm groups, while less (p<0.05) vlues were shown y 100 gm group These could minly relted to the effect of tnnins in cro pods on DM intke wheres less DM intke could found. Wghorn reported tht condensed tnnins could reduce fier, CP nd OM digestiility due to their inding properties nd inhiition of rumen microes nd lso they re ruminlly indigestile [60]. Consequently, digestiility of dietry components will e ffected nd lter the end products of fermenttion [37]. In this study digestiility of CP nd CF ws incresed with the supplementtion of cro pods (50 g/h/d). However, the ction of tnnins on nimls proly depends on their soluility, in the gstrointestinl trct [9] But this effect ws in certin level, s [52] nd [59] indicting no dverse effect of dietry inclusion of CT elow 5% level on nutrient intke nd utiliztion. This ws confirmed y the finding 275

4 Scientific Ppers Series Mngement, Economic Engineering in Agriculture nd Rurl Development in this study, for group fed 50 g/h/d. Group supplemented with 50gm cro pods hd the highest (P<0.05) totl digestile nutrients (TDN) nd digestile crude protein (DCP) vlues thn other groups. On the other hnd 100 gm s0upplement ws the less (P<0.05) vlue of TDN nd DCP (Tle 2). Other groups hd intermedite vlues with insignificnt effect (P>0.05). These results were greed with the finding of Dey [18]. Tle 2. Apprent digestiility coefficients, nutritive vlues nd nitrogen utiliztion of the experimentl diets Items Control Cro pods SEM P vlue DM intke (g/h/d): CFM RS Totl DMI, g Digestiility coefficients (%): DM c OM c CP c CF c EE c NFE c Nutritive vlues (%): TDN c DCP c Nitrogen utiliztion (g/h/d): N-intke (g/d) Urine-N (g/d) Fecl-N (g/d) c N-digested (g/d) c N-lnce (g/d) c N- lnce s % of N-digested c c mens in the sme row with different superscripts re significntly differ (P< 0.05). N-intke ws lower (P<0.05) for group dily received 100 gm/h, while other groups hd insignificnt differences. Although ll groups of nimls hd positive N lnce, those receive 100 gm/h/d ws the less (P<0.05) one. So, it ws reflected the worst utiliztion one. It is cler tht the group received 50 gm cro pods group ws the etter (P<0.05) one in their N utiliztion compred with ll groups. The less N utiliztion of tht group dily received 100 gm/d, could e relted to the tnnin effect on reducing digestiility of dietry proteins y their ility to form insolule complexes with proteins or digestive enzymes [10]. However, with incresing level of cro pods, NI decrese nd oth FN nd UN increse, which resulted in less N utiliztion. So, the more (P<0.05) CP digestiility nd N-utiliztion of rms dily supplemented with 50 gm./h, ws within the level suggested y Bhtt et l. (2000) [11] nd Dey et l. (2008) [19] s it ppered from the increse numer of cteri in the rumen s well. Ruminl fermenttion Ruminl ph vlues were not significntly ffected y different levels of cro pods supplementtion (Tle 3). So, no ny phenomenon of cidosis ws noticed, s cro pods hve potentility to prevent ruminl cidosis y reducing rpid strch hydrolysis. Concentrtion of ruminl metolites (NH3-N nd VFA s) ws significntly (P< 0.05) vried mong the experimentl rtions. Supplementtion with 100 mg cro pods hd the lower NH3-N nd TVFA s concentrtions; while 50 mg ws recorded the highest vlue of TVFA s compred with other groups. However, tnnins hd limited effects on the vlues of ph, VFA production ut resulted in noticele decrese in NH3. Molr proportion (%) of propionic (P) cid ws not 276

5 Scientific Ppers Series Mngement, Economic Engineering in Agriculture nd Rurl Development significntly ffected y different levels of cro pods supplementtion, while cetic (A) cid, A:P rtio nd rumen volume (L) were higher (P<0.05) with 50 mg supplementtion. In the sme mne, Beuchemin reported tht incresing supplementtion levels of quercho CT resulted in decresing totl VFA concentrtion, ut in contrs cette nd A:P rtio were lso reduced [6]. On the other hnd, Crull oserved no chnge in totl VFA concentrtion in sheep supplemented with lck wttle tree CT, ut cette ws decresed, while propionte incresed [14]. Benchr nd Aguerre found tht totl concentrtions of VFA nd molr proportions of individul VFA were not ffected y feeding quercho CT [7] [1]. So, the effect of cro pods on totl VFA concentrtion nd VFA pttern hve een vrile mong studies depending on the dosge rte nd the consists of condensed tnnin in cro pods Tle 3. Effect of different levels of cro pods on rumen prmeters of Brki sheep Items Control source [38], [24] nd [41]. Microil nitrogen synthesis (MN) ws rnged from to (g/d) for 100 nd 50 mg supplementtion respectively, which reflected y the rte of out flow, s it considered s suitle for efficient ruminl cellulolytic cteri nd MN synthesis with the dose of 50 mg in this study. However, ruminl NH3- N levels for sheep fed 50 gm cro pods in this study, were higher thn the criticl level (25mg) suggested y Stter & Slyter [51] for microil protein synthesis. Supplementtion of cro pods significntly (P<0.05) decresed popultion of totl Protozo s compred with control. So, these results could end out tht cro pods could chnge popultion of rumen microorgnism. However, ruminl protozo count is vrile due to the tnnins effects s it hd een reported in mny ssys crried out in vivo [46]. Cro pods SEM P vlue ph NH 3-N concentrtion c (mg/100mlr.l) TVFA concentrtion c (meq/100 mlr.l) Acetic cid, % c propionic cid, % Butyric cid, % Acetic : propionic rtio Rumen volume (L) 3.11 c d 0.07 < Rte of out flow (%h) c 5.58 d Microil N yield (g /d) c d Totl cteri counts, c 0.04 < cfu/ml Totl protozo counts, 106 cfu /ml c ,, c nd d : mens in the sme row with different superscripts re significntly (P<0.05) different. Degrdtion kinetics It ws illustrted in (Tle 4) tht wshing loss frction for DM, nd CP mong groups ws insignificntly different (P> 0.05) mong groups. while, degrdle frction nd rte of degrdtion c ws higher (P<0.05) for the control group thn other groups. The effective degrdility ED of DM or CP ws lower (P<0.05) for group supplemented with 100 gm. cro pods. While, it ws higher for control group. These could e relted to the less nutrients digestiility of 100 gm cro pods group, nd s tnnins re cple in inding with dietry proteins, resulted in less degrdle nutrients in the rumen [9], [25], [45] nd [40]. Although, CT could decrese ruminl degrdility of CP, it increses mount of CP reches the omsums nd smll intestine [40].This could e confirmed in the present 277

6 Scientific Ppers Series Mngement, Economic Engineering in Agriculture nd Rurl Development study s RUP ws less (P<0.05) for group with 100 mg. cro pods. Tle 4. Degrdtion kinetics of DM nd CP in the rumen of sheep fed the experimentl diets Items Control DM Cro pods SEM P vlue A B c C c EDDM CP A B c C c EDCP c RUP c , nd c : mens in the sme row with different superscripts re significntly (P<0.05) different. ED: effective degrdility= + [ c/c + k], where k is pssge rte (ssumed to e 0.05/hr.). RUP = ED (Orskov nd McDonld, 1979). = solule frction (%). = potentilly degrdle frction (%). c = rte of degrdility (% h -1 ). RUP = ruminely undegrdle protein Growth performnce Highest (P<0.05) finl ody weight nd verge dily weight gin were recorded with lms dily fed 50 gm cro pods, while the lowest vlues were recorded with the group fed 100 gm. Other groups hd intermedite vlues without significnt differences (P>0.05). These could e due to the more feed intke of 50 gm cro pods followed y other groups nd the less intke of 100 gm group. Although tht dietry CT generlly tend to decrese DMI when it is in high level, ut there re exceptions [42]; [47], the first two reported tht no dverse effect on DMI, BW or ADG, while the second found tht there is n increse in DMI. So, the lst could e supported our finding with 50 gm supplementtion of cro pods. However, this ws reflected in more economic efficiency with 50 gm cro pods group. Tle 5. Growth performnce of lms fed experimentl diets Items Control Cro Pods SEM P vlue Initil ody weight, kg Finl ody weight, kg c Dily weight gin, g/d c Dily feed intke, g/d Feed conversion rtio c Economic efficiency: Averge dily feed cost (L.E) Price of dily gin(l.e) c 0.3T Economicl return (L.E /h/d) Economic efficiency (%) c Reltive economic efficiency c ,,c different letters within row denote significnt differences etween tretments (P<0.05). These results greed with the finding of Alonso-Diz who reported decline in ody weight gin of gots with incresing concentrtions of CT in their feed. He dded tht concentrtions of CT eyond 6% tended to relte negtively on ody weight gin 278 which supports with the oservtions of [2]. Lizrdo nd Andres-Elis reported tht dietry cro (contined CT) did not ffect the growth of wened piglets [36] nd [3]. However, mny mmmls, especilly rowsers, re le to produce proline-rich

7 Scientific Ppers Series Mngement, Economic Engineering in Agriculture nd Rurl Development slivry proteins (PRP) tht re le to in to dietry CT to inctivte them [5]. Blood prmeters The lood prmeters re presented in Tle 6. Dily supplementtion of cro pods (100 gm) ws resulted in less (P<0.05) lood glucose, totl protein (TP) nd ure-n, while other cro pods groups ws shown more (P<0.05) concentrtion of TP nd gloulin, this could e due to the feed intke nd digestiility coefficients for these groups. On the other hnd, control hd the more concentrtion of glucose nd cholesterol. Higher (P<0.05) concentrtion of lumin nd ure-n ws found for control nd tht supplemented with the less level of cro pods (25 nd 50 gm/h/d). However, ll groups hd insignificnt different for liver nd kidney functions (AST, ALT nd cretnine). So, nothing unexpected phenomen hppened in the present study. Tle 6. Blood serum iochemicl components of lms fed experimentl diets sitems Control Cro pods SEM P vlue Glucose (mmol/l) c Cholesterol (mmol/l) Totl protein (g/l) c Alumin (g/l) Gloulin (g/l) c Ure N (mmol/l) Cretinine (μmol/l) AST (IU/L) ALT (IU/L) , nd c: mens in the sme row with different superscripts re significntly (P<0.05) different. Antioxidnt enzymes ctivity Dt in Tle (7) indicted tht supplementtion with different levels of cro pods significntly (P<0.05) incresed SOD, CAT, GPx nd GST ctivities nd decresed TBARS in plsm compred to the control group. This reveled tht cro pods could prevent the lipid peroxidtion y enhncing the SOD, CAT nd GPx ctivities. So, the phenolic compounds in cro exert powerful ntioxidnt effects nd inhiit lipid peroxidtion y ntioxidtive enzymes [35], [34] nd [28]. The ntioxidnt ctivity cused y the presence of these compounds could hve dditionl effects, spring other ntioxidnts nd protecting molecules from oxidtive dmge during digestion nd preserving the intestinl epithelium from potentil oxidtive dmge cused y dietry fctors or cteril metolism [27]. Tle 7. Chnges in the plsm ntioxidnt ctivities in lms supplemented with different levels of cro (Certoni siliqu L) Items Control Cro pods SEM P vlue TBARS GPx GST CAT SOD nd : mens in the sme row with different superscripts re significntly (P<0.05) different. CONCLUSIONS From this study it cn e concluded tht dily supplement with Cro Pods especilly t 50 gm could improve the productive performnces of the growing Lms nd chieved etter feed efficiency, economic efficiency nd reltive economic efficiency. 279

8 Scientific Ppers Series Mngement, Economic Engineering in Agriculture nd Rurl Development Moreover, the hemtologicl nd iochemicl prmeters investigted in this study did not revel ny signs of illness in lms due to dily consumption of 50 gm Cro Pods. However it needs more investigtion for long term feeding in the field of eef or diry milk production. REFERENCES [1] Aguerre, M. J., Wttiux, M. A., Cpozzolo, M. C., Lencioni, P., Crl, C., 2010, Effect of querchochestnut tnnin extrcts t two dietry crude protein levels on performnce nd rumen fermenttion of diry cows. J. Diry Sci. 93(Suppl. 1):445. [2]Alonso-Díz, M.A., Torres-Acost, J.F.J., Sndovl- Cstro, C.A., Hoste, H., 2010, Tnnins in tropicl tree fodders fed to smll ruminnts: A friendly foe? Smll Ruminnt Reserch 89: [3]Andres-Elis, N., Pujols, J., Bdiol, I., Torrllrdon, D., 2007, Effect of nucleotides nd cro pulp on gut helth nd performnce of wenling piglets. Livest. Sci. 108: [4]AOAC, 2007, Officil Method of Anlysis. (18 th ed), Assocition of Officil Anlyticl Chemists. Wshington, DC, USA. [5]Austin, P.J., Suchr, L.A., Roins, C.T., Hgermn, A.E., 1989, Tnnins-inding proteins in sliv of deer nd their sence in sliv of sheep nd cttle. J. Chem. Ecol. 15, [6]Beuchemin, K. A., McGinn, S. M., Mrtinez, T. F., McAllister, T. A., 2007, Use of condensed tnnin extrct from quercho trees to reduce methne emissions from cttle. J. Anim. Sci. 85: [7]Benchr, C., McAllister, T. A., Chouinrd, P. Y., 2008, Digestion, ruminl fermenttion, cilite protozol popultions, nd milk production from diry cows fed cinnmldehyde, quercho condensed tnnin, or Yucc schidiger sponin extrct. J. Diry Sci. 91: [8]Bengly, S., Mhlongo, S., Nshli, I. V., 2007, The effect of wttle tnnin on intke digestiility, nitrogen retention nd growth performnce of gots in South Afric. Livestock Reserch for Rurl Development 19(4): Article 50. [9]Ben-Slem, H., Nefzoui, A., Ben-Slem, L., Tissernd, J.L., 1999, Different mens of dministering polyethylene glycol to sheep: effect on the nutritive vlue of Acci cynophyll Lindl. Folige Anim. Sci. 68: [10]Beutler, E., Dixon, O., Kelly, B.M., 1963, Improved method for the determintion of lood glutthione. J. L. Clin. Med. 61: [11]Bhtt, R., Krishnmoorthy, U., Mohmmed, F., 2000, Effect of feeding tmrind (Tmrindus indic) seed husk s source of tnnin on dry mtter intke, digestiility of nutrients nd production performnce of crossred diry cows in mid-lcttion. Anim. Feed Sci. Technol. 83: [12] Bigi, G., Cipollini, I., Pulicks, B.R., Roth, F.X., 2010, Effect of tnnins on growth performnce nd intestinl ecosystem in wened piglets. Arch. Anim. Nutr. 64:121. [13]Borhmi, B.E.A., Fhmy, W.G., El-Shzly, K., 1992, Rumen environment microil protein synthesis nd nitrogen lnce in sheep. In: A Proceeding of "Mnipultion of rumen micro-orgnisms". Inter. Confe. From Sept [14]Crull, J. E., Kreuzer, M. Mchmüller, A., Hess, H. D., 2005, Supplementtion of Acci mernsii tnnins decreses methnogenesis nd urinry nitrogen in forge-fed sheep. Aust. J. Agric. Res. 56: [15]Cheeke, P. R., Ptton, N. M., Tempelton, G. S., 1982, Rit production. 5 th Edition, Interstte Printers nd Pulishers, Dnville 11. [16]Custodio, L., Fernndes, E., Escp, A. L., Fjrdo, A., Aligue, R., Alericio, F., Neng, N. R., Nogueir, J. M. F., Romno, A., 2011, Antioxidnt nd cytotoxic ctivities of cro tree fruit pulps re strongly influenced y gender nd cultivr. J. Agric. Food Chem. 59: [17]Demeyerd, I., 1981, Rumen microes nd digestion of plnt cell wlls. Agriculture nd Environment, 6, [18]Dey, A., Dutt, N., Shrm, K., Pttnik, A. K., 2007, Livestock Reserch Rurl Development. 19 (12): 295. [19]Dey, A., Dutt, N., Shrm, K., Pttnik, A.K., 2008, Effect of dietry inclusion of Ficus performnce of lms. Smll Rum Res.,75: [20]Doums, B.T., Wtson, W.A., Biggs, H.G., 1971, Alumin stndrds nd mesurement of serum with romocresol green. Clin. Chem. Act., 31(1): [21]Duncn, D.B., 1955, Multiple rnges nd multiple F- test. Biometric 11: [22]El-Shzly, K. Ahmed, E.I.A., Ng, M.A., Borhmi, B.E.A., 1976, A clorimetric technique using chromium-ethylen dimins tetrcette for mesuring rumen volume. J. Agric. Sci. Cm., 87: 369. [23]FAO FAOSTAT sttisticl dtse. Rome (ville t fostt.fo.org). [24]Frutos, P., Hervás, G., Giráldez, F.J., Fernández, M., Mntecón, A.R., 2000, Digestive utiliztion of quercho-treted soy en mel in sheep. J. Agr. Sci., 134, [25]Frutos, P., Hervás, G., Giráldez, F.J., Mntecón, A.R., 2004, Review. Tnnins nd ruminnt nutrition. Spnish Journl of Agriculturl Reserch 2: [26]Gsmi-Bouker, A., Bergoui, R. Khldi, A. Mosquer-Losd, M. R., Kett, A., 2008, First ttempt to study cro pulp utiliztion in rits feeding. World J. Agric. Sci. 4(1): [27]Goñi, I., Serrno, J., 2005, The intke of dietry fier from grpe seeds modifies the ntioxidnt sttus in rt cecum. J. Sci. Food Agric., 85: [28] Hrorne, J.B., Bxter, H.,1999, The Hndook of Nturl Flvonoids, Vol 2. John Wiley nd Sons, New York. [29]Henry, R.J., Cnnon, D.C., Winkelmn, J.W., 1971, Clinicl chemistry: Principles nd Techniques.

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