Effects of Probiotic (Bacillus subtilis) on Laying Performance, Blood Biochemical Properties and Intestinal Microflora of Shaoxing Duck

Transcription

1 Interntionl Journl of Poultry Science 10 (): 53-59, 2011 ISSN Asin Network for Scientific Informtion, 2011 Effects of Proiotic (Bcillus sutilis) on Lying Performnce, Blood Biochemicl Properties nd Intestinl Microflor of Shoxing Duck Wei Fen Li, Imrn Rshid Rjput, Xin Xu, Y Li Li, Jin Lei, Qin Hung nd Min Qi Wng Key Lortory of Moleculr Animl Nutrition nd Feed Sciences, Ministry of Eduction, College of Animl Science, Zhejing University, Hngzhou, , P.R. Chin Astrct: The study ws conducted to evlute the effects of proiotic supplementtion (Bcillus sutilis) in diet on performnce, lood properties nd microflor of Shoxing duck. A popultion of 200 lying ducks (160 dys old) ws divided into two groups ech hving five replictions. The control (T0) were fed on sl diet while T1 with Bcillus sutilis 1 x 10 CFU/kg in ddition to sl diet for thirty five dys. The results showed tht for T1 group, the egg lying rte incresed (3.79%) significntly, while decrese of out 12.60%, 23.52% nd 40.14% ws oserved in egg triglyceride, totl cholesterol nd mlondildehyde, respectively. However, the lood glucose content nd lkline phosphtse ctivity incresed y 15.13% nd 53.41%, significntly. Moreover, there ws decrese of lood cholesterol, uric cid nd mmoni in T1 y 2.6%, 22.39% nd 62.5%, respectively. As compred to T0, T1 showed significnt incresed in the mount of cecl microflor, i.e. generl eroic cteri (1.6%) nd Lctocillus (9.92%). In conclusion, supplementtion of Bcillus sutilis in diets hd significnt effects on some spects of egg composition, iochemicl properties of lood nd few microflor of Shoxing ducks. Key words: Proiotics, Bcillus sutilis, totl cholesterol, shoxing duck MATERIALS AND METHODS The study ws conducted to investigte the effects of supplementtion (Bucillus sutilis) on the performnce, iochemicl chnges in lood nd cecum microflor of Shoxing Ducks. A totl of 200 shoxing ducks (160-dy- old t lying, (verge weigh 1.72±0.02 kg) were rndomly divided into two groups, with five replictions ech group nd 20 ducks per repliction. Ducks were rre; in stndrd frm with spreded litter (whet rown 2-3 inches thick) ducks hd free ccess of feed nd wter with fcility of plyground nd fresh wter pond. promoters, feed svers, nutritionl ioregultors nd Control group (T0) ws only fed on sic diet (Tle 1) help in improving the performnce nd helth (Vnell for 35 dys while treted group (T1) ws fed on sic diet et l., 1990; Jinet l., 199), y relesing of proteses, with Bcillus sutilis (1 x 10 cfu/kg). The nutrients in the INTRODUCTION Proiotics re living microes given orlly to proliferte in the Gstrointestinl Trct (GI) of the host nd crete eneficil conditions for nutrients utiliztion (Nhshon et l., 2005; Jin, et l., 1996; Jin et l., 2000). Proiotic produce positive lnce of digestive microflor nd limiting the dmge cused y pthogenic cteri, improves epithelil cell integrity nd incresed immune response (Vnell et l., 1990; Jin et l., 2000; Wenk, 2000; Pnd et l., 2001; Linge, 2005). However, if microorgnisms nd/or sustnces, which contriute to the proper microil lnce, were dded to the diet, the niml would continully receive oost to estlish the proper microil popultion. Mny investigtors otined dt in the literture relting to the use of cteril cultures to control nd/or promote the proper environmentl conditions for the estlishment of n idel microil popultion in n niml s digestive trct through ppliction of cultured cteri. Species tht hve trditionlly een regrded sfe proiotics re Enterococcus, Bifidocterium nd Bcillus (Ishishi nd Ymzki, 2001). Preventive ppliction of proiotics chieved etter utiliztion of nutrients nd they hve positive effect on environment of gstrointestinl trct (Cpcrov et l., 2009). These ct s growth et-mnnnse nd mny other enzymes, these germinting nd outgrowing spores ssist in the feed digestion process (Hooge, 2003). Severl studies demonstrted tht the supplementtion of proiotics to poultry diets incresed performnce of the irds, stilized the intestinl microil flor nd lso reduced incidence of disese (Smny nd Ymuchi, 2002; Hooge et l., 2004). Therefore keeping in view the importnce of the suject nd rpid scrce of wterfowls the present study hd een designed to evlute the effects of proiotic (Bcillus sutilis) on performnce, lood properties nd cecum microil popultion of the shoxing ducks. Corresponding Author: Min Qi Wng, Key Lortory of Moleculr Animl Nutrition nd Feed Sciences, Ministry of Eduction, College of Animl Science, Zhejing University, Hngzhou, , P.R. Chin 53

2 Tle 1: Ingredients nd nutrient levels of sl diet Ingredients Percentge Nutrients Content (%) Corn 46.5 DE (MJ/kg) Soyen mel 23.0 CP 1.00 Rpeseed cke.0 C 3.09 Whet flour 10.0 P 0.75 Mono clcium phosphte 1.5 Lys 0.95 Limestone 6.7 Met 0.35 Sodium chloride Premix compound Premix compound ech kilogrm contined: vitmin A, 5,000 IU; choleclciferol, 1,500 IU; tocopheryl cette, 11 IU; mendione, 1.1 mg; thimine HCl, 3.0 mg; rioflvin, 5.0 mg; pyridoxine HCl, 2.2 mg; cynocolmin, 0.66 meq; nicin, 44 mg; C pntothente, 12 mg; choline chloride, 220 mg; folic cid, 0.55 mg; D-iotin, 0.11 mg; Mn, 0.0 mg; Zn, 60.0 mg; Fe, 30.0 mg; Cu, 5.0 mg; I, 2.0 mg; nd Se, 0.15 mg. Premixes were formulted to meet recommended levels for minerls nd vitmins (NRC, 1994). Composition ws nlyzed y AOAC (1990) sl feed were nlyzed y the method of Numnn nd Bssler (1993) (Tle 1). Nutrient levels of the sl diet were nlyzed for CP, Lys, Met, C nd P y the methods of AOAC (1990). Smpling, testing nd oservtions: Feed consumption, Egg numer nd weight were recorded dily during the whole test period. Egg lying rte, men egg weight, dily egg mss, feed-egg rtio were determined using formul, 1, 2, 3 nd 4 respectively. Totleggnumer Egglyingrte = x100% Totl duck numer xdys Totleggmss Men egg weight = Totl eggnumer Dily eggmss = Egglyingrte xmen egg weight Totl feedconsumption Feed- eggrtio = Totlegg weight Collection of eggs ws performed weekly nd three eggs of ech replicte were rndomly selected to nlyze egg qulity for egg weight, shell thickness, horizontl-verticl rtio, yolk colour nd hugh unit. Concentrtions of triglyceride, cholesterol nd mlondildehyde in egg yolk were mesured using commercil kits (Roche Dignostics GmH, Mnheim, Germny) on iochemistry utonlyzer (RX Dyton, Shining Sun Technology, Beijing, Chin). At the end of experiment, three ducks with similr ody weight of ech replicte were selected, then weighed nd slughtered fter fsting for 12 h. The hert, liver, spleen, pncres of ech duck were collected, weighed nd kept t -0 C. Reltive orgn weight ws determined ccording to the following formul: reltive orgn weight = orgn weight/live ody weight. Moreover, After merciful killing, weight of internl orgns, such s liver, spleen, pncres nd hert ws recorded. Ten eggs from ech replicte were rndomly selected to collect the yolk. The egg yolk cholesterol ws extrcted y the method of Folch et l. (1956) s modified y (Wshurn nd Nix, 1974) nd their chemicl composition ws determined ccording to the methods of Assocition of Officil Anlyticl Chemists (AOAC, 1990). Cecl contents were lso collected for microil nlysis. Blood smples were collected from crotid rtery loodletting using 23 guge needle. Serum ws seprted nd purified y 10 minutes centrifugtion (5,500 x g), then spirted y pipette nd kept in 1.5 ml, ependrof tues t -0 C for further nlysis. Biochemicl lood indexes i.e. triglyceride, totl cholesterol, glucose, uric cid, serum mmoni, lkline phosphtse, glutmic oxlocetic trnsminse, Glutmic pyruvic trnsminse were lso nlyzed y RX Dyton uto nlyzer using (Roche Dignostics commercil) kits. Microil nlysis Preprtion of test smple: Cecum content (1 g) ws diluted in sterile diluents peptone wter solution (9 ml) -1-5 to mke primry dilution (10 ). Then series up to 10 dilution ws prepred y trnsferring primry dilution (1 ml) into test tue contining sterile diluents (9 ml) to -2 otin 10 dilution nd repeting the opertions with -2 sterile diluents (9 ml) using the 10 nd further dilutions to otin 10, 10 nd/or 10. The cecl content smples from ech group were used to enumerte the count of totl eroic cteri, totl neroic cteri, Bifidocterium, E. coli, Lctocillus nd Bcillus y selective culture medium fter collection of cecl contents Pre prepred test smple (1 ml) of 10, 10 nd/or 10 dilutions ws trnsferred into petri dishes contining selective medi in duplicte through dispensing pipette (100 µl) with sterile plstic tips nd wrm sterile gr medium (15 ml) ws mixed with inoculums. The mixture ws llowed to solidify nd incuted. Prllel to tht, control pltes were lso prepred using similr medium (15 ml) to check the sterility. Counting of cecl microflor: The dishes contining more thn 30 nd/or fewer thn 300 colonies were selected nd counted using colony counter. The result ws clculted using following formul: c N = (n + 0.1xn)d 1 2 3c : Sum of colonies counted on ll the dishes retined. n1 : Numer of dishes retined in the first dilution. n2 : Numer of dishes retined in the second dilution. d : Dilution fctor corresponding to the first dilution. 54

3 Tle 2: Effect of supplementl Bcillus sutilis on lying performnce of Shoxing ducks Group Live ody weight (kg) Egg lying rte (%) Men egg weight (g) Dily egg mss (g) Feed-egg rtio T ± ± ± ± ±0.039 T ± ± ± ± ±0.093 Mens in columns with different letters re significntly different from ech other (p<0.05) Sttisticl nlysis: Dt ws processed using SPSS 16.0 for Windows (SPSS Inc., Chicgo, IL). Piredsmples t test ws chosen s compre men to nlyze index differences etween group T0 nd T1. The proility of (p<0.05) ws considered s sttements of sttisticl significnce. RESULTS AND DISCUSSION The effect of supplementl proiotic Bcillus sutilis on the lying performnce of Shoxing ducks is presented in Tle 2. Live ody weight of irds ws not significntly improved comprtively to control group; results showed tht the supplementtion of 1 x 10 Bcillus sutilis/kg diet rought out significnt (p<0.05) improvements in intensity of egg lying in supplemented group s compred to control groups. However, slight numericl improvements were oserved in mens of dily egg collection, feed conversion rtio nd verge egg weight, when irds were fed with 1 x 10 Bcillus sutilis/kg diet. It is of interested to note tht egg weight did not decline y the increse of intensity of egg lying in Bcillus sutilis fed ducks. These results re in conformity with the findings of Zeweil nd Ismil (199) nd Adel- Azeem et l. (2005), who lso indicted tht egg production, egg weight nd egg mss were improved in lying hens fed proiotic-supplemented diets. The positive effects of Bcillus sutilis supplementtion to Shoxing ducks could e due to decrese in the multipliction of hrmful cteri resulting from improvement in gut environment nd enhnced nutrient utiliztion (Miles, 1993). Bcillus sutilis my, lso, enhnce enzymtic ctivity in the digestive trct resulting in improving nutrient utiliztion. The positive effects of proiotic supplementtion, oserved in the present study, re in greement, with y El-Sheikh (2006). The effects of the dietry supplementtion with proiotic on reltive weight of some internl orgns of Shoxing ducks re summrized in Tle 3. These results suggested tht the effect of proiotic supplementtion s feed dditive hs not significntly (p>0.05), ffective on liver, spleen, hert nd pncrese weights Tle 3: Effect of supplementl Bcillus sutilis on weights of selected orgns Orgns/(g) T0 T1 Liver 47.03± ±2.64 Spleen 0.53± ±0.11 Hert 10.36± ±0.53 Pncres 5.24± ±0.15 Mens in rows with sme letters re non-significnt (p>0.05). T0 = Control, T1 = B. sutilis treted Tle 4: Effect of supplementl Bcillus sutilis on egg qulity of Shoxing ducks Contents T0 T1 Egg weight (g) 63.26± ±1.57 Shell thickness (mm) 0.39± ±0.00 Horizontl-verticl 0.77± ±0.01 Egg yolk color 11.20± ±0.00 Hugh unit 2.16± ±2.12 Triglyceride (mmol/l) ± ±2.95 Totl cholesterol (mmol/l) ± ±2.29 Mlondildehyde ± ±39.99 Mens in rows with different letters re significntly different from ech other (p<0.05). T0 = Control, T1 = B. sutilis treted respectively. Jin et l. (199) nd Bozkurt et l. (2005) lso found tht dietry preiotic nd proiotic supplementtion did not stimulte the internl orgn weight of roilers. The present results re in disgreement with the findings of Hill et l. (1957), Visek (197), Henry et l. (196) nd Engerg et l. (2000) indicted tht dietry inclusion of feed grde ntiiotics, given s growth promoters, reduced liver, spleen nd intestine weight y thinning the intestinl wll evoked prticulrly y ntimicroil ctivity in gut lumen. However, series of reports suggested similr ntimicroil mode of ction for preiotics (Iji et l., 2001; Shne, 2001; Ferket, 2004), orgnic cids (Broek, 2000; Diner, 2004) nd proiotics (Vnell et l., 1990; Yeo nd Kim, 1997; Wenk, 2000). Moreover, definitive dt re lcking with respect to effects of dietry proiotics on the internl orgns of poultry. Anlysis of the egg qulity trits of shoxing ducks fter supplementtion of Bcillus sutilis, dt showed in Tle 4. Proiotic (Bcillus sutilis) inclusion did not influence significntly on the egg weight, shell thickness, horizontl-verticl egg yolk color nd hugh unit (p>0.05), re in greement with the findings of Cernigli et l. (193), Mohn et l. (1995), Hdddin et l. (1996) nd Chen nd Chen (2003). Complementry reports y the Nhshon et l. (1996) nd Hdddin et l. (1996) suggested tht dditionl concentrtion of iologicl dditives did not influence the egg qulity significntly (p>0.05). But, present results re entirely different s reported y Nhshon et l., 1992; Tortuero nd Fernndez, 1995, showed tht using vitl iomss of proiotic supplements ffects the egg qulity significntly (p<0.05) tht might e relted to the concentrtion, strin of cteri, induction root nd the form of ppliction (viility, dry or Liquid nd their products). Dmron et l. (1976) nd Jensen et l. (197) lso found significnt (p<0.05) improvements in interior 55

4 Tle 5: Effect of supplementl Bcillus sutilis on iochemicl prmeters of lood in Shoxing ducks Contents T0 T1 Glucose (GLU, mmol/l) 25.41± ±1.159 Triglyceride (TG, mmol/l) 4.6± ±0.345 Totl cholesterol (T-CHL, mmol/l) 3.435± ±0.299 Uric cid (UA, mg/l) ± ±1.69 Serum mmoni (SA, umol/l) ± ± GlutmicoxlocitteTrnsminse (GOT) ± ±1.54 Glutmic Pyruvic Trnsminse (GPT) ± ±0.92 Alkline Phosphtse (ALP/100 ml) ± ±4.57 Mens in rows with different letters re significntly different from ech other (p<0.05). T0 = Control, T1 = B. sutilis treted egg qulity s mesured y hough units in hens fed distillers feeds nd corn fermenttion solules. In some reports, presented y Nhshon et l. (1992) nd Tortuero nd Fernndez (1995) the dosges of proiotic nd concentrtion of cteri used in the diet produces significnt effects on egg qulity. In diets there re more cteri per grm of feed compring with Nhshon et l. (1996) nd Hdddin et l. (1996), 2200 mg/kg vs mg/kg; 10 cfu/gr feed vs cfu/g feed respectively. Thus, increse of qulity might e relted 9 to the vitl form with higher doses up to 10 cfu/g feed of proiotic. In ddition, proiotics hd significnt (p<0.05) effect on egg yolk cholesterol, shown in Tle 4. Hdddin et l. (1996); Sutton et l. (194) nd Mrks nd Wshurn (1991) lso reported tht inclusion of Lctocillus cidophilus in three ges (40, 44 nd 4 week) ffects egg cholesterol in 40 week of production not 44 nd 4. The results of present study reveled tht the dietry proiotic Bcillus sutilis suppressed the cteril growth tht produce urese which correlte to lood uric cid nd mmoni showed (Tle 5), significntly decresed in tretment group (p<0.05) s compre to control. A similr effect ws found when roiler chicks were fed diet supplemented with Lctocillus cidophilus (Tortuero, 1973; Frncis et l., 197). In the present study, depressed net mmoni production in lood contents y dietry supplementtion of proiotics (Tle 5) might e due to sorption of mmoni y Bcillus sutilis (Hedon nd Dwson, 1990) nd suppressing urese ctivity cn e eneficil for improving helth of wterfowls. Reduction of lood ure nd mmoni concentrtions due to supplementl dietry proiotics hs een reported in rts (Duffy nd Brooks, 199), poultry (Blog et l., 1994), rits (Hussin et l., 1996; Al-Br nd Al-Aghri, 1996) nd steers (Hussin nd Cheeke, 1995). However, Lowe nd Kershw (1997) oserved tht lood ure significntly incresed in the cts. The Tle 5 showed significnt (p<0.05) increse in glucose nd decese in totl cholesterol of lood. The present results nd the findings of Yoon et l. (2004); Al-Kssie nd Ad-Aljleel (2007) re similr. However, Kim et l. (2003) reported tht roiler fed on diet contined A. oryze cused reduction in cholesterol level of lood serum of roiler. Sme results were otined y Al-Sodini (2005) y using proiotic Aspergillus niger s feed supplement. Tle 6: Effect of supplementl Bcillus sutilis on cecl microe s concentrtion of Shoxing ducks Microes (cfu/g) T0 T1 Aeroic cteri.317± ±0.121 Aneroic cteri ± ±0.276 Bifidocterium ± ±0.565 E. coli.271± ±0.294 Lctocillus 7.977± ±0.244 Bcillus 9.624± ±0.24 Mens in rows with different letters re significntly different from ech other (p<0.05). T0 = Control, T1 = B. sutilis treted Addition of proiotics in feed improves the overll ody iochemicl fctors, ut stress of dditionl dietry supplement my e cuse of increse on glucose in lood. Triglyceride in lood did not show significnt difference in control nd tretment group. However, lkline phosphtse ws significntly higher in tretment group. No significnt difference ws oserved in glutmicoxlocitte trnsminse nd glutmic pyruvic trnsminse etween control nd tretment groups. The present results re in greement with Adel-Ftth et l. (200), who reported non significnt (p>0.05) difference in the SGPT nd SGOT levels. Appliction of Bcillus sutilis s feed supplement ffects the microes of cecum significntly (p<0.05) incresed the popultion of eroic microes nd lctocillus (Tle 6). The presented results re in greement with the findings of Dimcho et l. (2005). In ddition, no numericl improvement nd degrdtion ws oserved in cecum generl neroic cteri, Bifidoterium, E. coli nd Bcillus. These results resemle with Hristev et l. (2004) who found tht, proiotics did not influence on concentrtion of Eenterococci growth in Muskovy ducks. The results of current study gives line to oserve further study out the proiotics on qulity improvements. Moreover, Pndy et l. (2010) concluded tht dietry supplementtion of proiotics in the diet of poultry could e given without producing ny dverse effect. REFERENCES Adel-Azeem, F.A., G.M. Nemtllh nd F.A.A. Irhim, Effect of dietry protein level with some nturl iologicl feed dditives supplementtion on productive nd physiologicl performnce of Jpnese quil. Egy. Poult. Sci., 25:

5 Adel-Ftth, S.A., M.H. El-Snhoury, N.M. El-Medny nd F. Adel-Azeem, 200. Thyroid ctivity, some lood constituents, orgns morphology nd performnce of roiler chicks fed supplementl orgnic cids. Int. J. Poult. Sci., 7: Al-Br, A.M. nd A.M. Al-Aghri, Influence of deodorse in comintion with different levels of protein on rit feed intke, ody weight nd utiliztion of ure. 6th World Rit Cong., Toulouse, Frnce, 1: Al-Kssie, G.A.M. nd R.A. Ad-Aljleel, Effect of Glli cid Irq proiotic on some hysiologicl properties nd intestinl microflor in roiler chicks. Proceeding of second Veterinry, Scientific Conference. College Vet. Med. Univ. Bghdd, pp: Al-Sodini, A.A.H.K., Using the Aspergillus niger s preiotic in rtion of roiler Ross. Strin. Thesis of M.Sc College of Agriculture, Bghdd University of Irq. AOAC, Officil methods of nlysis, Officil Anlyticl Chemists, 15th Edn., Arlington. Inc, USA. Blog, J.M., N.B. Anthony, C.W. Wll, R.D. Wlker, R.C. Rth nd W.E. Huff, Effect of urese inhiitor nd ceiling fns scites in roiler. 2-Blood vriles, scites scores nd ody nd orgn weights. Poult. Sci., 73: Bozkurt, M., K. Kucukyilmz, A.U. Ctl nd M. Cinr, Growth performnce nd crcss yield of roiler chickens given ntiiotic, mnnn oligoscchride nd dextrn oligoscchride supplemented diets. Nutrition Biotechnology in the Feed nd Food Industries. Proc. 21st nnul symp. Lexington, Kentucky, USA., pp: 69. Broek Vn Den, G. Ir., Orgnic cid: Nturl link etween drug nd growth promoter. Feed Mix (Specil), pp: Cpcrov, M., J. Kovcik nd M. Mellen, Chnges of iochemicl prmeters poultry lood fter ppliction of proiotic preprte. Slovk Univ. Agric., Nitr, pp: Cernigli, G.J., A.C. Goodling nd J.A. Herert, 193. The response of lyers to feeding Lctocillus fermenttion product. Poult. Sci., 62: Chen, Y.C. nd T.C. Chen, Effects of commercil proiotic or preiotic supplementtion on production, size nd qulity of hen s egg. Poult. Sci., 2 (Suppl. 1): 330 (Astr). Dmron, B.L., A.R. Eldered nd R.H. Hrms, An improvement in interior egg qulity y the feeding of rewers dried grins. Poult. Sci., 55: Diner, J., Orgnic cids: Cn they replce ntiiotic growth promoters? Feed Int., 25: Dimcho, D., B. Svetln, S. Tsvetomir nd V. Ttin, Effect of feeding lctin proiotic on performnce, some lood prmeters nd cecl microflor of mule ducklings. Trki J. Sci., 3: Duffy, C. nd Brooks, 199. Using yucc schidiger in pig diets: Effects on nitrogen metolism. Proc. Alltech s 14th Annu. Symp. Nottinghm University Press, Nottinghm, UK., pp: 61. El-Sheikh, H.M.M., Effect of proiotic supplementtion to roiler chicken diets on its performnce, crcss chrcteristics nd lood constituents, M.Sc. Thesis, Fculty of Agriculture (S Bsh), Alexndri Uni, Egypt. Engerg, R.M., M.S. Hedemnn, T.D. Leser nd B.B. Jensen, Effect of zinc citrcin nd slinomycin on intestinl microflor nd performnce of roilers. Poult. Sci., 79: Ferket, P.R., Alterntives to ntiiotics in poultry production: Responses, prcticl experience nd recommendtions nutritionl iotechnology in the feed nd food industries. In: Proc. Alltech s 20th Annu. Symp. Nottinghm University Press, pp: Folch, J., M. Less nd G.H. Slone Stnley, A simple method for the isoltion nd purifiction of totl lipid from niml tissues. J. Bio. Chem., 226: Frncis, C., D.M. Jnky, A.S. Arf nd R.H. Hrms, 197. Interreltionship of lctocillus nd zinc citrcin in the diets of turkey poults. Poult. Sci., 57: Hdddin, M.S.Y., S.M. Adulrhim, E.A.R. Hshlmoun nd R.K. Roinson, The effects of Lctocillus cidophilus on the production nd chemicl composition of hen's eggs. Poult. Sci., 75: Hedon, R.D. nd K.A. Dwson, Yucc extrct controls tmospheric mmoni levels. Feedstuffs, 62: Henry, P.R., C.B. Ammermn nd R.D. Miles, 196. Influence of virginimycin nd dietry mngnese on performnce, mngnese utiliztion nd intestinl trct weight of roilers. Poult. Sci., 65: Hill, C.H., A.D. Keeling nd J.W. Kelly, Studies on the effects of ntiiotics on the intestinl weights of chicks. J. Nutr., 62: Hooge, D.M., Bcillus spores my enhnce roiler performnce. Feed Stuffs, 75: Hooge, D.M., H. Ishimru nd M.D. Sims, Influence of dietry Bcillus sutilis C-3102 spores on live performnce of roiler chickens in four controlled pen trils. J. Applied Poult. Res., 13: Hristev, H., A. Bochukov nd G. Penchev, Comprtive study on the effect of Lctin proiotic on some microil nd histologicl chrcteristics of the digestive trct of Muscovy ducklings. J. Cent. Eur. Agric., 4:

6 Hussin, I., A.M. Ismil nd P.R. Cheeke, Effects Nhshon, S.N., H.S. Nkue nd I.W. Mirosh, of feeding yucc schidiger extrct in diets vrying in crude protein nd ure contents on growth performnce nd cecum nd lood ure nd mmoni concentrtions of rits. Anim. Feed Sci. Technol., 62: Performnce of single com white leghorn fed diet supplemented with live microil during the growth nd egg lying phses. Anim. Feed Sci. Technol., 57: Nhshon, S.N., H.S. Nkue nd L.W. Mirosh, Hussin, I. nd P.R. Cheeke, Effect of dietry Effect of direct-fed microils on nutrient retention Yucc schidiger extrct on rumen nd lood nd production prmeters of lying pullets. Poult. profiles of steers fed concentrte-or roughgesed diets. Anim. Feed Sci. Technol., 51: Sci., 71(suppl.1): 111(Astr). Nhshon, S.N., H.S. Nkue nd L.W. Mirosh, Iji, P.A., A.A. Ski nd D.R. Tivey, Intestinl Performnce of Single Com White Leghorn lyers structure nd function of roiler chickens on diets fed with live microil during the growth nd egg. supplemented with mnnn oligoscchride. J. Trki J. Sci., Vol. 3. Sci., Food Agric., 1: Numnn, C. nd R. Bssler, Chemicl nlyses Ishishi, N. nd S. Ymzki, Proiotics nd of feed stuff. Method ook III. (3rd Edn.), VDLUFAsfety. Am. J. Clin. Nutr., 73: Press. Drmstdt, Germny. Jensen, L.S., C.H. Chng nd S.P. Wilson, 197. Interior egg qulity: Improvement y distillers feeds nd Pnd, A.K., M.R. Reddy nd N.K. Prhrj, trce elements. Poult. Sci., 57: Dietry supplementtion of proiotic on serum Jin, L.Z., Y.W. Ho, A.M. Adullh nd S. Jlludin, cholesterol nd gut microflor of roilers. In. J. Effects of Lctocillus culture on the digestive Anim. Sci., 71: enzymes in chicken intestine. Proc. th Animl Pndey Anju, Milind M. Nik nd S.K. Duey, Science Congress, Tokyo, Chi, Jpn, pp: 224- Orgnic metolites produced y Virio 225. Jin, L.Z., Y.W. Ho, A.M. Adullh nd S. Jlludin, Digestive nd cteril enzyme ctivities in roiler fed diets supplemented with Lctocillus cultures. Poult. Sci., 79: Jin, L.Z., Y.W. Ho, N. Adullh, M.A. Ali nd S. Jlludin, 199. Effects of dherent Lctocillus cultures on growth, weight of orgns nd intestinl microflor nd voltile ftty cids in roilers. Anim. Feed Sci., 70: Kim, S.H., S.Y. Prk, D.J. Yu, S.J. Lee, K.S. Ryu nd D.G. prhemolyticus strin An3 isolted from Gon mullet inhiit cteril fish pthogens. Afr. J. Biotechnol., 9: Smny, M. nd K. Ymuchi, Histologicl ltertions of intestinl villi in chickens fed dried Bcillus sutilis vr. ntto. Comp. Bio. Physio., 133: Shne, M.S., Mnnn oligoscchrides in poultry nutrition: mechnism nd enefits. In: Proc. Alltechs 17th Ann. Symp., Science nd Technology in the Feed Industry. Eds Lyons, T.P. nd Jcques, K.A., Lee, Effect of feeding Aspergillus oryze Nottinghm University Press, pp: ferments on performnce, intestinl microflor, Sutton, C.D., W.M. Nuir nd G.E.J. Mitchell, 194. lood serum components nd environmentl Cholesterol metolism in lying hen s influenced fctorsin roiler. Koreon J. Poult. Sci., 30: y dietry cholesterol, cloric intke nd genotype. Linge, P., The use of proiotics nd yest Poult. Sci., 63: derivtes in Indi. World Poult., 21: Tortuero, F. nd E. Fernndez, Effect of Lowe, J.A. nd S.J. Kershw, The meliorting inclusion of microil culture in rley-sed diets effect of Yucc schidiger extrct on cnine nd fed to lying hens. Anim. Feed Sci. Technol., 53: feline fecl rom. Res. Vet. Sci., 63: Mrks, H.L. nd H.W. Wshurn, Plsm nd yolk Tortuero, F., Influence of implnttion of cholesterol levels in Jpnese quil diver gently selected for plsm cholesterol response to Lctocillus cidophilus in chicks on the drenocoticotropin. Poult. Sci., 70: growth, feed conversion, ml sorption of fts Miles, R.D., Mnipultion of the microflor of the syndrome nd intestinl flor. Poult. Sci., 52: gstrointestinl trct: Nturl wys to prevent coloniztion y pthogens. In: Biotechnology in the Vnell, N., E. Teller nd M. Focnt, Proiotics in Feed Industry. Proc. of Alltech's Ninth Annul Symposium, T.P. Lyons, Ed. Alltech Technicl Pulictions. Nicholsville, Ky, USA., pp: Mohn, B., R. Kdirvel, M. Bhskrn nd A. Notrjn, Effect of proiotic supplementtion on serum/ yolk cholesterol nd on egg shell thickness in lyers. Br. Poult. Sci., 36: niml nutrition: A review. Arch. Anim. Nutr., 40: Visek, W.J., 197. The mode of growth promotion y ntiiotics. J. Anim. Sci., 46: Wshurn, K.W. nd D.F. Nix, A rpid technique for extrction of yolk cholesterol. Poult. Sci., 53:

7 Wenk, C., Recent dvnces in niml feed Yoon, C., C.S. N, J.H. Prk, S.K. Hn, Y.M. Nm nd J.T. dditives such s metolic modifiers, Kwon, Effect of feeding multiple proiotics on ntimicroil gents, proiotics, enzymes nd performnce nd fecl noxious gs emission in highly ville minerls. Asin-Austr. J. Anim. Sci., roiler chicks. Koren J. Poult. Sci., 3: : Zeweil, H.S. nd F.S.A. Ismil, 199. Effect of proiotic or Yeo, J. nd Kyu-Il Kim, Effect of feeding diets herl feed dditives on the production nd egg contining n ntiiotic, proiotic, yucc extrct on qulity of lying Jpnese quil hens. J. Agric. Sci., growth nd intestinl urese ctivity in roiler 23: chicks. Poult. Sci., 76: