A.A. Ogunjobi, A.T. Adesoji, O.J. Babayemi and S. Omotosho. Department of Botany and Microbiology, University of Ibadan, Ibadan, Nigeria 2

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1 Worl Journl of Agriulturl Sienes 8 (4): , 2012 ISSN IDOSI Pulitions, 2012 DOI: /iosi.wjs Effets of Comine Strters Culture of Ltoillus plntrum on Fermenttion Qulity, Aeroi Stility n Aeptility y Ruminnt of Pnium mximum Silge A.A. Ogunjoi, A.T. Aesoji, O.J. Byemi n S. Omotosho 1 Deprtment of Botny n Miroiology, University of In, In, Nigeri 2 Deprtment of Animl siene, University of In, In, Nigeri Art: This uy exmine e effets of two L. plntrum rters t re physiohemilly ifferent on 6 Pnium mximum silge fermenttion. The grss ws inoulte wi pproximtely 10 fu/g of e rter ulture. Aeroi ility, hemil omposition n eptility of e silge proue y ruminnt were monitore t 15 ys intervls for 60 ys. The uies oserve rpi rop in e ph of e inoulte silge to 3.90 t y 30, inrese in e ount of lti i teri (LAB) from 9.00 log fu/g t y 15 to log fu/g t y 60 while erese in spoilge miroorgnisms like mol from 4.09 log fu/g t y 15 to log fu/g t y 60 in e inoulte silge. The rue protein (CP) of e silge ws lso oserve to inrese from 6.56 g/100 DM on fresh wilte grss to 9.63 g/100 ry mtter (DM) t y 30 n 45 in e inoulte silge while e neutrl etergent fier (NDF) n i etergent fier (ADF) of e silge erese in e inoulte silge. The NDF erese from g/100 DM t y 15 to g/100 DM on y 60. The inoulte n e ontrol silge were not eroilly le when expose to ir t e 45 n 60 y of ensiling. The spoilge orgnism like ye inrese from 4.34 log fu/g t y 1 of exposure to 4.4 log fu/g t e seon y of exposure t e 45 ys of ensiling. The We Afrin wrf sheep use in is uy showe low eptility to e inoulte silge n e ontrol silge wi COP of 0.98, 0.90, 0.81 n 0.58 t ys 15, 30, 45 n 60 respetively ompre to e uninoulte ontrol whih hs oeffiient of performne (COP) of 1.01, 1.10, 1.18 n 1.42 t ys 15, 30, 45 n 60 respetively. Key wors: Strter ulture Silge Spoilge Miroorgnisms Ltoillus plntrum Sheep INTRODUCTION pollutnt to e environment n re regre s nturl prouts [4]. Mny types of homofermenttive lti i The preservtion of forge rops s silge epens teri (LAB) hve een ommerilly prepre s silge on e proution of suffiient i to inhiit tivity of itives. These selete rins enourge rpi ph unesirle miroorgnisms uner neroi onitions. eline n inhiit Dry Mtter loss s well s lorii The epiphyti lti i teri (LAB) present on forge fermenttion, erey improving preservtion in silo [5]. rops onvert sugr into lti i in e ensiling On mny frms, silge is often expose to vrying proess. As result, e ph is reue, n e forge is egrees of eroi eteriortion efore n uring preserve. However, when e silo is opene, eroi feeing. For exmple, e inility to remove suffiient onitions previl uring feeing time. The silge is quntities of silge from silos etween feeings n result sujete to eroi miroil grow n is potentilly in prolonge exposure to ir. Also,poorly pke silge is unle [1-3]. noer ommon reson for silges spoiling efore In orer to improve e ensiling proess, vrious feeing. In n ttempt to erese lor, some prouers hemil n iologil itives hve een evelope. hoose to fee eir heifers or eers one every 2 or 3 The iologil itives re vntgeous euse ey ys. When silge is purhse n move y truks, re sfe, esy to use, non-orrosive to mhinery, non- silge my sit on n open p n expose to ir for Corresponing Auor: A.A. Ogunjoi, Deprtment of Botny n Miroiology, University of In, In, Nigeri. 436

2 Worl J. Agri. Si., 8 (4): , 2012 severl ys. Silos t re not well pke n sele Preprtion of Silge Inoulnts: Ientifie n n expose re prtiulrly sujete to eroi hrterize Ltoillus plntrum from Pnium eteriortion. Fees t hve unergone eroi mximum silge from Environmentl n Biotehnology eteriortion hve eir nutritionl vlue reue n LAB, Deprtment of Botny n Miroiology, University present hzrs to e environment (isposl of spoile of In, In, Oyo Stte, Nigeri ws inoulte into fee). Therefore, improving e eroi ility of silges 500mL of prepre MRS ro in Erlenmeyer s flsk. oul onfer suntil vntge to prouers [6]. It ws lter inute on shker inutor for 2-3 ys Mny reserhers hve propose wys to improve e t 250rpm. After whih ws ilute wi 2500 ml of erile eroi ility of silges. For inne, pplying iille wter. Plte ount on MRS gr ws rrie out propioni i-se itives hve enhne e eroi y pipetting 1ml into e gr plte. ility of orn silge [7]. However, suh itives re oly n us miroil inoulnts re suitute s Sheule of Ensiling Experiment: Pnium mximum eonomil lterntives. Also, ing of enefiil grss t ws elish t e tehing n reserh frm miroorgnisms (miroil inoulnts) to fees prior to of e university of In, In, Oyo Stte, Nigeri ensiling improves fermenttion [8]. L. uhneri hs een ws hrvee in e mon of Novemer, 2008 fter Six mrkete in omintion wi homolti i teri, weeks of re-grow. The grss ws hoppe etween 2- whih re ommonly e to forge to inrese lti 3 m n wilte for 24 h. Smples of e wilte grss i proution, rpi rop of ph, n to improve DM ws tken for hemil n miroiologil nlysis. reovery [9]. However, ere re no long term uies on The hoppe grss ws en ivie into equl portion of e effets of omintion of two physiohemilly 1kg n pke into polyene gs n properly sele ifferent homolti i teri on silge. to exlue ir for neroi onition fter inoultion The im of is uy is to etermine e effets of 6 wi pproximtely 10 fu/g of e L. plntrum. inoulting physiohemilly ifferent isoltes of The silge ws opene fter 15, 30, 45 n 60 ys for L. plntrum on P. mximum silge. It lso exmines its miroiologil, hemil, n eptility y ruminnt. effet on fermenttion, eroi ility n eptility The eroi ility te ws rrie out t 45 n 60 y ruminnt nimls. ys of ensiling. There ws ontrol i.e. uninoulte silge t eh of e y in whih e silge ws opene. MATERIALS AND METHODS The experiment ws replite 3 times to reue error. Sreening of Isoltes: The ulture meium use for e Miroil, Physil n Chemil Anlysis of e Silge: sreening ws MRS ro. The orgnisms were ultivte Extly 1 g of e silge ws mixe wi 9 ml of erile t ifferent temperture of 25 C, 35 C, 37 C, 40 C n 45 C iille wter t eh of e ys of opening e in ro. The orgnisms were lso ultivte in ro silge. This ws for seril ilution etween 10 6 to prepre into vrious ph of 2, 4, 6 n 8 in miro The ilutions were en plte on e following mei: erophili environment. The orgnisms were grown in De-Mn-Rogos-Shpe gr (MRS) to etet LAB, eroi environment i.e. in n inutor t 37 C n Nutrient Agr (NA) to etet eroi teri, Ye neroi environment i.e. miroerophili environment in extrt Agr (YEA) for eteting ye, Potto Dextrose ro to etermine e effets of oxygen on e isoltes. Agr (PDA) to etet Moul n Eosin Meylene Blue The orgnisms were grown in ifferent MRS ro (EMB) for oliforms. Coliforms were ultivte 37 C for prepre t ifferent NCl onentrtion of 3%, 5%, 7% 1-2 ys. LAB ws ultivte t 37 C in miroerophili n 9%. After 48 h of grow of e isolte ulture, environment for 2 ys. The eroi teri were lso grow ws monitore wi spetrophotometer ultivte t 1-2 ys t 37 C, Moul n Ye were (SpetrumL 752S). The isolte ulture ws ultivte ultivte t 25 C for 3 ys. Numers of olony forming in MRS ro for 48hrs n e entrifuge t 3000 rpm for (fu) were expresse in log fu/ml/g. 15 min. The superntnt ws en titrte wi 0.25 mol Temperture of e silge ws mesure wi NOH n 1mL of phenolphlein initor (0.5% in ermometer. After whih 25 g of e silge ws tken 50% lohol). The titrtle iity ws lulte s from e silge t eh of e ys of opening n mixe lti i (%). The titrtle i ws lulte wi 100 ml of iille wter. It ws shken on shker oring to A.O.A.C [10]. t 250 rpm for 15 min n filtere wi filter pper. 437

3 Worl J. Agri. Si., 8 (4): , 2012 The filtrte ws use for e etermintion of e ph wi ph meter. DM i.e. ry mtter ontent of e silge were etermine y oven rying t 80 C for 48 h. Ash, rue fire, eer extrt ontent of e silge ws etermine oring to A.O.A.C. [10]. Neutrl etergent fire (NDF), Ai etergent fire (ADF) n Ai etergent lignin (ADL) were etermine oring to Goering n Vn Soe, [11], Crue protein (CP) ontent of e silge ws lso etermine y e Kjelhl meo [10]. Aeroi Stility Te: At 45 n 60 ys of ensiling, e silge ws sujete to eroi ility te t l for 7 ys. Smples of e silge were ple in petri ish n ple in esitors. Smples were tken t y 0, 2, 5, n 7 for e miroiologil nlysis of e silge. Moreover, e ount of e LAB, ye, moul, eroi teri n oliforms were etermine s mentione ove. Aeptility y Ruminnt: Eight We Afri wrf sheep weighing kg n out two yers ol were use to evlute e free hoie intke of e silge (ontrol n LAB trete) t eh ys of termintion of e experiment i.e. 15, 30, 45 n 60 ys respetively. In o e LAB trete n e ontrol silge ple in rtegi feeing troughs mesuring 1 m x 2 m, 2.5 kg eh of e silge ws ple on it. The sheep were en llowe to fee for 6 h per y on eh y of e fee out. Consumption ws mesure y eution of remnnts from e mount of silge serve fter 4 h. The silge preferre ws ssesse from e oeffiient of preferene (COP) vlue, lulte from e rtion etween e intke of e iniviul silge trete wi LAB or ontrol ivie y e verge intke of e silge trete wi LAB or ontrol [12] on is sis, silge tretment ws inferre to e reltively eptle if COP ws greter n unity. Sttiil Anlysis of Silge: The t were nlyze y nlysis of vrine n men were seprte y Dunn s multiple rnge [13]. RESULTS Physiohemil Sreening of LAB Isoltes: After e physiohemil sreening of ifferent (eighteen) isoltes of Ltoillus plntrum n two isoltes of L. revis. Two L. plntrum were selete for e inoultion of e P. mximum silge. The results of ese tes were shown in Tles 1, 2, 3 n 4 respetively inite Tle 1: Grow of LAB in MRS ro t ifferent temperture etermine y optil ensity Orgnisms 25 C 35 C 37 C 40 C 45 C All e numers on e orgnisms olumn represent L. plntrum exept numers 33 n 45 whih represent L. revis Tle 2: Optil ensity grow of LAB isolte in MRS ro t ifferent ph Orgnisms ph 2 ph 4 ph 6 ph All e numers on e orgnisms olumn represent L. plntrum exept numers 33 n 45 whih represent L. revis 438

4 Worl J. Agri. Si., 8 (4): , 2012 Tle 3: Optil ensity grow of LAB isoltes in MRS ro t ifferent NCl onentrtion Orgnisms 3% 5% 7% 9% All e numers on e orgnisms olumn represent L. plntrum exept numers 33 n 45 whih represent L. revis Tle 4: Quntity of lti i ( ) proue n grow of LAB isoltes in ifferent O regime 2 Orgnisms Titrle lti i Orgnisms Aneroi Aeroi All e numers on e orgnisms olumn represent L. plntrum exept numers 33 n 45 whih represent L. revis Ltoillus plntrum (i.e. Org 60) grow over wie rnge of ph hving e highe grow optil ensity of t ph 8 n optil ensity of t ph 2 mong e isoltes sreene. Org 19 proue e highe onentrtion of lti i of in MRS ro. Therefore, ese two L. plntrum were selete for e inoultion. Miroil Suession in Silge t Different Dys of Ensiling: The result of miroil suession in silge t ifferent ys ws shown in tle 5. It ws oserve t, e spoilge orgnisms in e silge were inhiite in e silge. The totl eroi teri erese to 3.04 log fu/g t en of e 60 y of ensiling from 5.48 log fu/g t y 15 of ensiling. Ye ount in e inoulte silge ws lso oserve to e inhiite ut e ontrol silge ws lower t y 15 n 30. The ount erese from 5.3 log fu/g on e fresh wilte P. mximum to 4.00 log fu/g t y 15 n 30. It ws oserve t, e moul ount ws lower in e ontrol silge wi erese from 5.00 log fu/g on fresh wilte grss to 3.46 log fu/g in e ontrol silge t y 15 while 4.09 log fu/g ws oserve in e inoulte silge t e sme y. But lesser ount of 3.48 log fu/g, 3.56 log fu/g, 2.84 log fu/g t y 30, 45, n 60 respetively ws oserve in e inoulte silge. The LAB ount in e silge i not show signifint ifferene (p<0.05) t y 15 while signifint ifferene (p<0.05) ws oserve t y 30, 45 n 60 respetively. The oliform ount in e silge i not show signifint ifferene (p<0.05) t y 30 of ensiling ut signifint ifferene (p<0.05) ws oserve t e oer ys of ensiling. The oliform ount in e inoulte silge rnge from 4.00 log fu/g to 4.71 log fu/g while t in e ontrol silge rnge from 3.65 log fu/g to 4.30 log fu/g. Physil Assessment n Chemil Composition of e Silge t Different Dys of Ensiling: These results of e physil n hemil omposition of e silge t ifferent ys of ensiling were shown in Tles 6 n 7. The olour of guine grss (silge) in ll e LAB trete n untrete i.e. e ontrol silge vries from olive-green to greenish yellow t 15, 30, 45 n 60 ys of ensiling respetively. The smell/tes of 15 ys silge were mil fruity smell to shrp fruity smell for ll orgnism inoulte silges. Mil vinegr smell to shrp n very shrp vinegr oor/tes were pereive for e guine grss (ontrol silge) n ll e inoulte silge from ys 30 to 60 respetively. The ruture of guine grss silge for ll e silge were firm, visile n seprle in e overll silge roughout e entire perio of ensiling. 439

5 Worl J. Agri. Si., 8 (4): , 2012 Tle 5: Miroil suession in silge t ifferent ys of ensiling n miroil ount on fresh unensile wilte P. mximum (log fu/g) Dys of ensiling Totl eroi teri Control Totl eroi teri ount on fresh grss 3.6 Coliform Control Coliform ount on fresh wilte P. mximum 5.0 LAB Control LAB ount on fresh wilte P. mximum 5.0 Ye Control Ye ount on fresh wilte P. mximum 5.3 Moul Control Moul ount on fresh wilte P. mximum 5.0 Lti i teri (LAB), represent silge trete wi omintion of e two L. plntrum Vlues rry ifferent letters were signifintly ifferent t p < 0.05 Tle 6: Physil qulity of silge y olour, ruture n te of silge Tretments Dys ensile Prmeters Control 15 Colour Olive green Olive green Smell/tes Mil fruity smell Shrp fruity smell Struture Firm n seprle Firm n seprle 30 Colour Olive-green Olive-green yellow Smell/tes Vinegr Very shrp vinegr Struture Firm n seprle Visile n seprle 45 Colour Olive green Olive green Smell/tes Very shrp vinegr Very shrp vinegr Struture Visile, firm n seprle Seprle n visile 60 Colour Olive green Olive green Smell/tes Mil vinegr Very shrp vinegr Struture Visile n seprle Visile n seprle is e silge inoulte wi e omintion of e two L. plntrum Signifint ifferene (p<0.05) etween e ontrol inrese in e DM of e silges (inoulte n n inoulte silge ws oserve in e ph of e silge uninoulte) ompre to e DM of e fresh wilte t y 15 n 30 respetively. But no signifint grss. ifferene ws oserve in e ph t y 45 n 60 of The CP of e silge inrese from 6.56 g/100 DM ensiling. The ph of e ontrol silge ws 4.73 t y 30 oserve in e fresh wilte grss to 9.63 t y 15 n 30 while t of e inoulte silge ws 3.90 t y 30. in e inoulte silge whih erese to 8.53 g/100 DM No signifint ifferene (p>0.05) t y 15 ws n 8.31 g/100 DM t y 45 n 60 respetively. In e oserve in e temperture of e silge t y 15, 45 n uninoulte silge e CP of e silge inrese up to 60. At y 30 where signifint ifferene (p<0.05) ws 7.86 g/100 DM t y 45 ut erese to 6.78 g/100 DM oserve e temperture of e ontrol silge ws 28 C t y 60. while t of e inoulte silge ws C. Signifint ifferene (p<0.05) ws oserve in e Signifint ifferene (p<0.05) ws not oserve in Etylene extrt( EE) in ll e ys of ensiling etween e e DM of e silge t y 15 n 30. But ere ws ontrol n inoulte silge. 440

6 Worl J. Agri. Si., 8 (4): , 2012 The ADF ontent of e silge ws oserve to e Miroil Suession in Silge fter Exposure to Air signifintly ifferene (p<0.05) etween e ontrol n t 45 Dys of Ensiling: The results of e miroil inoulte silge in ll e ys of ensiling. The ADF ws suession fter exposure of e silges to ir t oserve to e lower (36.00 g/100 DM t y 30 n 45) in 45 ys of ensiling were shown in Tle 8. The e inoulte silge ompre to e uninoulte ount of e LAB ws oserve to erese from ontrol (37 g/100 DM t y 30 n 45). Similrly tren 9.74 log fu/g t e fir y of exposure to ws lso oserve in e NDF vlue in ll e silge. n 7.24 log fu/g t e 2 y n own to 3.44 log fu/g t The ADL of e ontrol silge t y 15 ws 7.89 y 7. Derese ws lso oserve in e ontrol g/100 DM while t of e inoulte silge ws 8.57 silge n erese to 3.64 log fu/g ws oserve g/100 DM n inrese to 8.60 g/100 DM t y 60 ws on 7 y of exposure. oserve in e ontrol while inrese to 8.82 g/100 DM The ye in e inoulte silge inrese rilly ws oserve t y 60 in e inoulte silge. n to 4.4 log fu/g t 2 y of exposure from 4.34 log fu/g The L. plntrum i not hve ny effet on e t e 1 y Inrese to 5.88 log fu/g ws oserve t rue fier (CF) of e silge euse signifint e 5 y of exposure. In e ontrol silge similr ifferene (p<0.050) ws not oserve in e silge inrese ws lso oserve ut 2.17 fu/g ws oserve (ontrol n inoulte) t ll e ensile ys. n in e ye ount on e 2 y. Tle 7: Physil n hemil omposition of silge t ifferent ys of ensiling Dys of ensiling Tretments ph Control ph of fresh wilte grss 6 0 Temperture ( C) Control DM (g/100g DM) Control DM of fresh wilte grss CP (g/100g DM) Control CP of fresh wilte grss 6.56 EE (g/100g DM) Control EE of fresh wilte grss Ash (g/100g DM) Control Ash of fresh wilte grss 8.00 ADF (g/100g DM) Control ADF of fresh wilte grss NDF (g/100g DM) Control NDF of fresh wilte grss ADL (g/100g DM) Control ADL of fresh wilte grss CF (g/100g DM) Control Dry mtter (DM), Crue protein (CP), Eer extrt (EE), Ai etergent fire (ADF), Neutrl etergent fire (NDF), Ai etergent Lignin (ADL), Crue fire (CF), ws silge inoulte wi omintion of e two L. plntrum. Vlues rry ifferent letters were signifintly ifferent t p <

7 Worl J. Agri. Si., 8 (4): , 2012 Tle 8: Miroil suession in silge fter exposure to ir t 45 ys of ensiling (log fu/g) Dys of exposure to ir Tretments Aeroi teri Control LAB Control Ye Control Moul Control Lti i teri (LAB), is silge inoulte wi e omintion of e two L. plntrum Vlue rry ifferent letters were signifintly ifferent t p < 0.05 Tle 9: Miroil suession in silge fter exposure to ir t 60 ys of ensiling Dys of exposure to ir Tretments Aeroi teri Control LAB Control Ye Control Moul Control Lti i teri (LAB), is silge inoulte wi omintion of e two L. plntrum. Vlue rry ifferent letters were signifintly ifferent t p < 0.05 Tle 10: Aeptility of orgnism trete n untrete P. mximum silge y We Afrin wrf sheep Dys of ensiling n eptility Tretment MDI COP MDI COP MDI COP MDI COP Control (MDI)(kgDM): men y mtter intke n (COP) oeffiient of preferene. is e silge inoulte wi e omintion of e two L. plntrum The moul of e silge inrese from 4.27 log fu/g exposure of e silge inrese in ll e silge. t e 1 to 5.90 log fu/g t 5 y in e ontrol silge, The ye inrese from 4.34 log fu/g t 1 y to while inrese from 3.56 log fu/g to 4.98 log fu/g t log fu/g t 5 y in e inoulte silge, while y ws oserve in e inoulte silge. n erese to 2.17 log fu/g t 2 y from 4.16 log fu/g t 1 y ws oserve in e uninoulte silge. Miroil Suession in Silge fter Exposure to Air t n The moul ount erese to 2.10 log fu/g t 2 60 Dys of Ensiling: The results of e miroil y from 4.27 log fu/g t1 y in e ontrol silge. suession t 60 ys of ensiling were shown in Tle 9. Inrese from 3.56 logfu/g t 1 y to 3.94 log fu/g t Spoilge miroorgnisms like ye, moul n eroi n 2 y n 4.98 log fu/g t 5 y ws oserve in e teri were oserve to inrese s e ys of inoulte silge. 442

8 Worl J. Agri. Si., 8 (4): , 2012 The eroi teri inrese from 4.94 log fu/g t silge ores well [18]. Therefore, in is uy, e ount 1 y to 9.84 log fu/g t 5 y in e ontrol silge, of LAB on e fresh grss ws low while t of e ye while inrese to 5.99 log fu/g t 5 y from 3.94 log ws high. But it ws oserve t s e ys of ensiling fu/g t 1 y ws oserve in e inoulte silge. inrese e ount of e LAB inrese t y 15 whih The LAB ount erese from 8.46 log fu/g to 4.87 erese t y 30 n inrese gin t y 45 n 60 log fu/g t 5 y in e ontrol silge, while erese to respetively in e inoulte silge. At 30 y of 4.99 log fu/g t 5 y from 9.74 log fu/g t 1 y ws ensiling it ws oserve in is uy t e ount of oserve in e inoulte silge. LAB ws higher in e ontrol silge n in e inoulte silge. This my e s result of ompetition Aeptility of Inoulte n Uninoulte P. etween e miroil inoulnts n e epiphyti LAB Mximum Silge y We Afrin Dwrf Sheep: The results of e eptility te re shown in Tle 10. Dt in tle inite t e sheep use onsume more of e silge from e ontrol silge n t of e inoulte silge. Hene, s e ys of ensiling inrese e COP of e inoulte silge erese while t of e ontrol silge inrese. The COP rnge etween 1.01 n 1.42 for e ontrol silge while t of e inoulte silge rnge etween 0.58 n 0.98 for ll e ensile ys. The MDI for e ontrol silge rnge etween 2.02 n 2.20 while t of e inoulte silge rnge etween 0.84 n DISCUSSION In orer to improve silge qulity, mny LAB-ontining iologil itives hve een evelope n re urrently ville [3, 14, 15]. These inoulnts my inhiit e grow of hrmful teri n enhne lti i fermenttion uring ensiling uie perios. The nturlly ourring LAB on plnt influenes e tion of silge inoulnts euse e introue teri mu ompete wi ese LAB [16]. Lin et l. [16] lso reporte t epiphyti LAB ply mjor role in silge fermenttion, n e numers of LAB hve eome signifint ftor in preiting e equy of silge fermenttion n etermining wheer to pply silge teril inoulnts. Among epiphyti LAB, lti i-prouing oi, e.g., Streptooi, Leuonoos, Peiooi, Ltooi, n Enterooi, rt ltte prouing fermenttion proess in silge, reting n eroi environment suitle for e evelopment of Ltoilli, lough it ws shown t ey grew vigorously only in e erly ge of ensiling proesses [17]. In ontr wi ese lti i-prouing oi, ltoilli ply n importnt role in promoting lti i fermenttion for longer time. Epiphyti ltoilli ounts on silge rops re usully low n vrile when 5 e ltoilli reh level of t le 10 fu/g of FM or it my e euse e two inoulnts re ntgonizing eh oer in e sme eologil environment sine two miroil inoulnts wi two ifferent physiologil properties re eing omine togeer. Previous uy wi e inoultion of miroil inoulnts use in is uy singly i.e. not in omintion shows shrp inrese in e ount of LAB s e ys of ensiling inrese wi rpi inhiition of spoilge orgnisms like eroi teri, ye, n Moul [19]. But e results in is uy ontrit e previous results euse s e ys of ensiling inrese ere ws no rpi erese in e ount of spoilge orgnism espeilly ye. This onform to report of Kung et l. [20] t inoultion oes not ffet e ount of ye n moul in whole high moiure orn silge (HMCS) when inoulte wi L. uhneri (6.6 x10 5 fu/g of HMCS ut when e pplition rte ws 4x10 5 fu/g of HMCS, 6x10 fu/g of HMCS n 8x10 fu/g of 5 5 HMCS, lso ey reporte t e ount of ye n moul ws reue ompre to e uninoulte silge. Hene, in is uy e ount of e spoilge orgnism my e reue if e miroil ount use for e inoultion of e silge is inrese or erese. Nevereless, rpi erese in e ount of ye is not n eviene of poor fermenttion sine e ount of LAB oserve in is uy is more n wht Hellings et l., [18] suggee shoul e e ount of LAB in silge wi goo fermenttion. All silge expose to ir eventully eteriortes s result of eroi miroil tivity [21]. The ftors t influene eteriortion inlues oxygen (exposure time), omposition of e miroil popultion, surte type, n temperture [21]. Ltte-ssimilting ye (Shromyes, Cni, Cryptoous, n Pihi spp.) re usully e initil use of eroi eteriortion [6]. Some rins of ltte-ssimilting ye grow well in n environment in whih e ph level is etween 3 n 8 n e temperture is uner 40 C [21]. These re onitions ommon in orn silge when it is fir expose 443

9 Worl J. Agri. Si., 8 (4): , 2012 to ir. As lti i n oer resiul sugrs re omue n ssimilte y ye, e temperture rts to rise [21]. One e temperture is ove 45 C, e mount of ye present elines n oer miroil orgnisms (illi) egin to umulte. As eroi eteriortion tkes ple, ere re hnges in e hemil prmeters of silge [22]. The ph level tens to inrese, mmoni n mines umulte, n e levels of orgni is (lti n eti i) ten to eline [21]. In is uy, it ws lso oserve t s e y of exposure of e silge to ir inrese e ount of spoilge orgnism like ye, eroi teri n moul inrese in e silge. The ount of e LAB whih is responsile for e proution of lti i erese s e time of exposure inrese in e silge. This proly oul hve resulte in e erese in e ph of e silge n erese in e temperture of ll e silge s ll ese proesses ourre in e silge. Homofermenttive teri, suh s Ltoillus plntrum, Ltoillus sei, Peioous spp. n Enteroous spp., minly proue lti i. Heterofermenttive teri, suh s Ltoillus uhneri, proue lti i, eti i, enol, n ron ioxie. Generlly, lti i is preferre in e silo euse it is ronger i n eti i [23]. Lti i reues ph fer, erey reuing plnt respirtion n enzyme tivity n inhiiting oer teri. However, eti i is etter inhiitor of ye, n mintins etter eroi ility n lti i. This ws similr to wht ourre in e LAB inoulte silge wi omintion of L. plntrum in is uy wi rpi reution of e ph of e silge ompre to t of e uninoulte silge. The min purpose of heterofermenttive inoulnts is to improve eroi (e presene of oxygen) ility y reuing e level of ye in e silge (high levels of ye n use heting). Ltoillus uhneri is e min heterofermenttive LAB use in forge rops in e U.S. [23]. Ltoillus uhneri proues more eti i n homofermenttive teri [24]. Therefore, in is uy espite e omintion of e two physiohemilly ifferent isoltes of L. plntrum e eroi ility of e inoulte n e uninoulte silge ws very poor wi inresing e ount of spoilge miroorgnisms wiin short perio of time. The inoulte silge ws even oserve to e less le n e ontrol silge t rily inrese in e ount of e moul n ye t e seon y of exposure whih ws not oserve in e ontrol silge. This ws similr to e report of Jonsson, [21] t some rins of ltte ssimilting ye grow well in n environment in whih e ph level is etween 3 n 8. Similr report ws lso reporte in e uies of Moon, [25] n Woolfor [26] t e i tolernt ye t ssimilte ltte (n usully not teri) re primrily responsile for e spoilge when expose to ir. The uors went furer to report t lti i y itself is not n effetive ntimyoti gent. However, in is uy, ere ws proly to e enough lti i in e silge t e y 45 n 60 of ensiling in e inoulte silge n e ontrol silge reily ville s surte for e ye. Kung n Rnjit [27] reporte t e inoulnts ontining len of homolti teri, propioniteri, n enzyme improve silge fermenttion y using more extensive homolti fermenttion n resulte in silge wi lower ADF n NDF ontent. This ws similr to wht ws oserve in is uy in whih silge inoulte wi e omintion of e two physiohemilly ifferent L. plntrum resulte in silge wi reue ADF n NDF ontent. Wheres, Aesogn et l. [28] in eir own uy reporte t e ermugrss silge ADF n NDF ws not ffete wi tretment wi inoulnts ontining mixture of P. pentoseu, L. uhneri n et-glunse, lph-mylse, n xylnse ontrry to e finings of is uy. Kung n Rnjit [27] lso reporte t eir inoulnt i not ffet e CP of eir silge whih ws similr to e report of Kleinshmit n Kung [24] in eir uy wi orn silge inoultion wi L. uhneri n Peioous pentoseus R1094 ut ontrry to e report of is uy. Therefore, e omintion of inoulnts use in is uy ws le to inrese e nutritionl qulities of e silge y reuing e NDF n ADF n inresing e CP of e silge. Silge is n importnt soure of lti i for e omei ruminnt. Cttle my e mintine entirely on silge n e intke of lti i y ese nimls my mount to 1 kg per y. It is iffiult to evlute e utiliztion of is frtion for proution. A numer of trils hve een rrie out ompring milk proution or weight gins, or o, in ruminnts fe silge n hy me from e sme soure of plnt mteril. Results from ese trils hve een vrile [29-35] ut, in generl, e slightly smller intke of ry mtter when silge ws fe ws offset y higher proution per unit of ry mtter. The net result of proution from nimls fe silge ws 444

10 Worl J. Agri. Si., 8 (4): , 2012 equl to or slightly greter n proution from ose fe 3. Weinerg, Z.G., G. Ashell, Y. Hen n A. Azrieli, hy. Therefore, e uninoulte silge oserve to e The effet of pplying lti i teri t well epte in is uy oul hve een of vntge ensiling on e eroi ility of silges. Journl of in t e sheep oul hve goo proution n Applie Bteriology, 75: feeing on inoulte silge. The reson why e nimls 4. Fily, I., et l., Silj klitesinin onsume less of e inoulte silge ompre to e rtt r lms n yeni geliþmeler. Interntionl uninoulte ontrol oul not e sertin ut it hs Animl Nutrition Congress 2000, s, een shown t e infusion of lti i into e rumen Isprt (in Turkish) [36, 37], or supplementing e rtion wi lti i 5. MDonl, P., A.R. Henerson n S.J.E. Heron, [32, 38-41], my use epression in voluntry intke The Biohemiry of Silge. Chlome of ry mtter in nimls fe hy [32, 37, 38, 39, 42]. Pulitions, Welton, Linoln, UK. The epression ws usully less n t oserve when 6. Rnjit N.K. n Kung L., Jr The Effet of silge ws fe, lough e mount of lti i e Ltoillus uhneri, Ltoillus plntrum, or ws similr to e quntities whih woul e ingee Chemil Preservtive on e Fermenttion n from feeing silge. However, Rooke [43] lso suggee Aeroi Stility of Corn Silge. Journl of Diry t e lti i my hve iret effet on pltiility, Siene, 83: sine sour te hs een ssoite wi reue 7. Kung, L., Jr., A.C. Sheper, A.M. Smgl, pltility. Any iret effets of lti i on K.M. Enres, C.A. Bessett, N.K. Rnjit n silge DM intke my e more importnt in e short J.L. Glney, The effet of preservtives se term n e long term n relte to negtive feek on propioni i on e fermenttion n eroi [44]. Therefore, it my e e presene of high ility of orn silge n totl mixe rtion. onentrtion of lti i s result of rpi Journl of Diry Siene, 81: fermenttion of e silge t ws responsile for low 8. Kung, L., Jr., R.S. Tung n K. Miorowski, eptility of e silge. Effet of miroil inoulnt n/or glyopeptie ntiioti on fermenttion n eroi ility of CONCLUSION wilte lflf silge. Animl Fee Siene Tehnology, 35: The omintion of e physiohemilly ifferent 9. Kung, L., Jr., M.R. Stokes n C.J. Lin, Silge L. plntrum ws effetive in rpily eresing e NDF itives. In Silge Siene n Tehnology. n ADF of e silge while e CP of e silge ws lso D.R. Buxton, R.E. Muk, n J.H. Hrrison, (es.). inrese. But it ws oserve t ensiling for more n Amerin Soiety of Agronomy, In., Crop Siene 45 ys le erese in e CP for e inoulte silge. Soiety of Ameri, Pulitions, Mison, WI, We, erefore, sugge t in orer to mintin goo pp: protein qulity of e silge, silge inoulte wi 10. A.O.A.C Offiil meos of nlysis of e omintion of ese two L. plntrum shoul not e AOAC, 15 e. AOAC, Arlington, Virgini, USA. fermente up to 45 y efore feeing out to nimls. 11. Goering, H.K. n P.J. Vn Soe, Forge fier nlyses. Agriulture Hnook, Nr. REFERENCES 379, Wshington 12. Byemi, O.J., F.T. Ajyi, A.A. Tiwo, 1. Ohym, Y. n S. Hr, Grow of yes M.A. Bmikole n A.K. Fjimi, Performne of isolte from silges on vrious mei n its We Afrin wrf gots fe Pnium mximum reltionship to eroi eteriortion of silge. Jpn n onentrte iets supplemente wi ll Journl of Zootehnology Siene, 46: (Ll purpureus), leuen (Leuen 2. Ohym, Y., S. Mski n S. Hr Ftors leuoephl) n Gliriii (Gliriii sepium) influening eroi eteriortion of silges n folige. Nigeri Journl of Animl Proution, hnges in hemil omposition fter opening 33(1): silos. Journl of Siene n Foo Agriulture, 13. Dunn, D.B., Multiple Rnge F-Te Biometry, 26: :

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12 Worl J. Agri. Si., 8 (4): , Emery, R.S., L.D. Brown, C.P. Huffmn, T.R. Lewis, 42. Bentley, O.G., R.R. Johnson, G.W. Royl, J.P. Everett n C.A. Lssiter, Comprtive F. Deerge, L.E. Kunkle, W.J. Tyznik n D.S. Bell, Feeing Vlue of Lti Ai n Grin for Diry Stuies on e Feeing Vlue of Aeti, Cttle. Journl of Animl Siene, 30: Propioni n Lti Ais wi Growing- Fttening 39. Thoms, J.W., L.A. Moore, M. Okmoto n Lms. Ohio Agrulturl Experiment Sttion, J.F. Sykes, A Stuy of Ftors Affeting Rte Reserh Bulletin 774, Wooer, OH, USA. of Intke of Heifers Fe Silge. Journl of Diry 43. Rooke, J.A., The effet of inresing iity or Siene., 44: osmollity of grss silge y e ition of free or 40. Rloff, H.D. n Shultz, L.H Some Effets of prtilly neutrlize lti i on silge intke y Feeing Lttes to Diry Cows. Journl of Diry sheep n upon osmollity n i-se lne. Siene, 46: Journl of Animl Siene, 61: Ekern, A. n J. Trei, Effiieny of 44. Offer, N.W., A omprison of e effets on energy utiliztion y young ttle ingeing voluntry intke y sheep of ietry ition of iets of hy, silge n hy supplemente eier silge juies or lti i solutions of e wi lti i. Journl of Diry Siene, sme neutrlizing vlue. Journl of Animl Siene, 46(6): 522-9, June. 64: