Int J Enteric Pathog May;5(2):35-40

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1 Interntionl Journl of Enteric Pthogens Int J Enteric Pthog My;5(2): Originl Article doi /ijep Supplementtion of Blend of Beneficil Bcteri nd Antiodies on Growth Performnce, Intestinl Mucos Morphology nd Right Hert Filure of Jpnese Quil (Coturnix coturnix jponic) Mohmmd Hsn Mehrei Hmzekolei 1, Adol Krim Zmni Moghddm 1 *, Seyed Sttr Tohidifr 1, Hjr Prskiy 1, Ali Heidri 2 1 Deprtment of Clinicl Sciences, Fculty of Veterinry Medicine, Shhrekord University, Shhrekord, Irn 2 Grduted Student of Veterinry medicine, Shhrekord University, Shhrekord, Irn *Corresponding Author: Adol Krim Zmni Moghddm, Deprtment of Clinicl Sciences, Fculty of Veterinry Medicine, Shhrekord University, Shhrekord, Postl Box: 115, Irn. Tel: ; Fx: ; Emil: zmni@vet.sku.c.ir Pulished Online Mrch 26, 2017 Keywords: Jpnese quil, Proiotic, Growth performnce, Right hert filure, Intestinl morphology Astrct Bckground: Erly nutrition of chicks with eneficil cteri might help in occupying the inner surfce of the intestinl trct. Interference of pthogens in intestinl microiot is well known s rrier effect, cteril interference, nd competitive exclusion. Ojectives: It ws hypothesized tht competitive exclusion in Jpnese quils with lend of eneficil cteri (Aqulend Avin proiotic) would enhnce quils growth performnce nd intestinl mucosl morphology. Furthermore, the study ws performed t 2100 m ove se level t Shhrekord University, so nother hypothesis ws the cpility of the proiotic for inhiiting right hert filure. Mterils nd Methods: One hundred fifty-six Jpnese quils were divided into 4 groups: 2 groups (Aqulend nd control) t stndrd environmentl temperture nd 2 (Aqu-stress nd Contstress) t cold-hypoxic environmentl sitution. Aqulend groups received the proiotic in the first 3 dys of life in drinking wter (0.5 g/100 irds/dy). Results: Feed conversion rtio (FCR) ws significntly reduced t the end of the experiment (dy 35) in oth Aqulend nd qu-stress groups compred to control nd cont-stress groups, respectively (P < 0.05). Crcss chrcteristics improvements in treted quils were not significnt. However, slight improvements in crcss nd spleen reltive weights were oserved (P > 0.05). Cont-stress group hd higher RV: TV rtio (0.28) nd heterophil: lymphocyte (H: L) rtio (1.22) thn qu-stress group: (0.25) nd (1.20), respectively (P > 0.05). Dt regrding to intestinl mucos morphology ws controversil ut the proiotic ws le to elevte duodenum villi surfce (P < 0.05) nd lso jejunum nd ileum lmin propri thickness. Conclusion: Otined dt suggests tht ddition of Aqulend Avin proiotic in the first 3 dys of life my improve growth performnce nd some intestinl mucos chrcteristics of Jpnese quils. Moreover, the proiotic might reduce right hert filure nd stress induced y cold-hypoxic sitution. Received Decemer 28, 2016; Revised Ferury 27, 2017; Accepted Mrch 7, 2017 Bckground Jpnese quil (Coturnix coturnix jponic) hs een considered s n importnt lortory niml nd lso n economiclly importnt egg nd met-type poultry worldwide. 1 However, stressful conditions cuse gret economic losses when lrge-scle rering fcilities hve een utilized for the quils. 2 Stressful sitution often depends on the region of rering; for instnce, cold-hypoxic sitution is one of such stressful situtions which occur in some mountinous res like Chhrmhl v Bkhtiri, which is the highest province of Irn. It could e stted tht environmentl ir chnges, like cold-stress is one of the most significnt dverse fctors tht might significntly influence metolism nd susequently the production of roiler chickens. 3 In iotic conditions to control the infectious nd in prticulr metolic diseses nd to improve the performnce, ntiiotics hve een used widely y producers 4 which hve their dverse effects in met qulity nd sfety. On the other hnd, cold-hypoxic environment could hsten the prevlence of right hert filure nd scites syndromes prticulrly in oilers nd to lesser degree in quils, resulting in reduced growth Copyright 2017 The Author(s); Pulished y Alorz University of Medicl Sciences. This is n open-ccess rticle distriuted under the terms of the Cretive Commons Attriution-NonCommercil 4.0 Interntionl License ( which permits copy nd redistriute the mteril just in noncommercil usges, provided the originl work is properly cited.

2 performnce nd high mortlity in irds. 5 Erly nutrition of chicks with eneficil cteri might help the intestinl trct in occupying the inner surfce of gut. Interference of pthogens in the intestinl microiot is well known s rrier effect, cteril interference, nd competitive exclusion. 6 Mechnisms tht re involved in the inhiition of pthogens consist of competing for nutrients, production of toxic compounds nd conditions (cteriocins, low ph, nd voltile ftty cids), competing for ttching to the sites of gut epithelium, nd stimulting the immune system. 7-9 Inoculting 1-d old roilers with eneficil cteri nd preiotics is effective for improving resistnce to pthogenic cteri nd intestinl function. 10,11 Aqulend Avin proiotic is lend of eneficil cteri nd ntiodies including Lctocillus, Streptococcus nd Bifidocterium strins which provides roilers with essentil microflor nd specific ntiodies ginst predominnt poultry pthogens. 12 Ojectives To the est of our knowledge, the effects of lends of eneficil cteri on right hert filure in cold stress sitution were not evluted in Jpnese quil; so the study imed to figure out the effects of Aqulend Avin proiotic ( lend of eneficil cteri nd ntiodies) on the growth performnce, intestinl mucos morphology, heterophil: lymphocyte (H: L) rtio nd right hert filure in Jpnese quils rered in norml nd cold hypoxic environment. Mterils nd Methods Animls, Mngement nd Tretments The experiment ws performed in experimentl poultry frm of Shhrekord University, Irn, which is locted t 2100 m ove se level. Quils were kept, rised, treted nd trnsported sed on the pproved rules for the humn tretment of nimls ccepted y the Institutionl Animl Cre nd Use Committee of Shhrekord University. One hundred fifty-six dy-old Jpnese quils (Coturnix coturnix jponic) from oth sexes were divided into 12 floor pens. Ech pen mesured 1.4 m 2 (13 quils/pen) nd ell drinker nd feed trough ws provided in ech pen. For mking deep litter system wood shvings were used s the litter mteril. Twenty-three hours of light ws ville for the irds in the first 7 dys nd then decresed to 20 hours t the end of the experiment for ll groups. At dy 1, environmentl temperture ws 36ºC nd ws grdully reduced to 24 C t the end of the experiment for norml temperture groups. At the end of the second week, 2 groups were trnsferred to nother re with the environmentl temperture of C till the end of the experiment in order to induce cold-hypoxic sitution. Quils were divided into 4 groups (3 replictes/ group), with 39 irds in ech group. Tretments were designed s follows: Group 1 (Aqulend): quils were fed sl corn soy en mel sed diet (Tle 1), nd received 0.5 g/100 irds/dy (0.065 g/replicte/dy) of Aqulend Avin (Agrnco Corp., Gles Interntionl Plz 2655 S Le Jeune Rd., Suite 805 Corl Gles, Fl USA), during the first 3 dys of life, in the drinking wter. Group 2 (control): irds were fed sl corn soy en mel sed diet, ut without ny proiotic ddition in the drinking wter. Group 3 (qu-stress): the group received the sme mount of Aqulend Avin s group 1, ut ws trnsferred to cold environment t the end of the second week. Dietry formul ws the sme s others. Group 4 (cont-stress): quils were fed the sme sl corn soy en mel sed diet without inclusion of proiotic, ut were sujected to stressful sitution of cold environment from dy 15. Mesurements Body weight (BW) gin nd food intke were mesured weekly in ech pen, s shown in the entire tril. Feed conversion rtio (FCR) dt were lso clculted nd corrected to exclude mortlity BWs in the entire tril. On dy 35th, 12 irds/tretment were selected rndomly for lood collection. Blood smples (3 ml) were collected from the rchil vein nd one drop ws smered on slide. Approximtely, fter 2-4 hours of fixtion with methyl lcohol, the smers were stined y My- Grünwld nd Giems stins. 13 A 100 grnulr nd nongrnulr leucocytes, were mesured on the slide using light microscopy, nd H:L rtios were determined. Moreover, fter leeding, the selected quils were euthnized for crcss processing. Assessed dt were hot eviscerted crcss, liver, spleen nd lung weight. To clculte right to totl ventriculr weight rtio (RV: TV rtio), herts were lso hrvested nd the ventricles were dissected nd weighed seprtely. The intestinl morphometric prmeters including Tle 1. Ingredients of the Experimentl Diet (g/kg) Ingredients Corn-Bsed Diet Corn seed Soyen mel Soyen oil 20.6 Di-clcium phosphte 8.3 Clcium cronte 12.6 Slt 1.6 Minerl-vitmin premix 5 DL-methionine 1.4 Vitmin D 3 1 Vitmin E c 1.5 Ech kilogrm of the minerl-vitmin premix contins vitmin A IU; vitmin D IU; vitmin E 1600 mg; vitmin K 3 35 mg; vitmin C 1200 mg; vitmin B 1 30 mg; vitmin B mg; vitmin B mcg; nicotinic cid 1300 mg; pntothenic cid 225 mg, choline chloride 8200 mg, iotin 3300 µg; nd Mn 1200 mg; Zn 1000 mg; Fe 1800 mg; Cu 400 mg; Se 8 mg; iodine 38 mg; nd C 180 g. Ech kg of vitmin D3 (choleclciferol) contins IU. c Ech kg of vitmin E (lph-tocopherol) contins IU. 36 Interntionl Journl of Enteric Pthogens Volume 5, Issue 2, My 2017

3 villus sizes (length, width, surfce re nd lmin propri thickness) were ssessed in the duodenum, jejunum nd ileum of ech euthnized quil t the end of the experiment. Briefly, fter fixtion of the mentioned segments of intestine in Clrk solution, sections were stined y periodic cid Schiff (PAS) regent, nd then rows of villi were seprted in thickness of the sections, trnsferred over glss slides nd covered with coverslip. Eyepiece grticules (10 ) nd mgnifiction of 100 were used in microscope to mesure villus sizes in these smples. The villus length nd width were used to clculte villus surfce re ccording to formul 2π (villus width/2) (villus length), s descried y Skmoto et l. 14 The spce etween se of the villus nd top of the musculris mucos ws mesured to find out the lmin propri thickness. Sttisticl Anlysis Results were nlyzed using SigmPlot 12.0 pckge. One-wy nlysis of vrince (ANOVA) test ws used to understnd the significnt differences mong groups nd P vlues < 0.05 were considered s significnt differences. Results There ws significnt decrement in FCR in Aqulend group compred to other groups t fourth nd fifth weeks (P < 0.05) which mens lower costs per ech kilogrms of met production. Additionlly, qu-stress group gined etter FCR thn cont-stress groups in fourth (P < 0.05) nd 5th (P > 0.05) weeks. Moreover, in the lst 2 weeks, FCR of control nd qu-stress groups were significntly (P < 0.05) lower thn cont-stress group (Tle 2). Crcss nd orgns reltive weights were not significntly different t the end of the experiment mong 4 tretments (Tle 3). However, quils in Aqulend group hd slightly higher percentge of crcss nd spleen weights reltive to BW (P > 0.05). Figures 1, 2 nd 3 mnifest villus surfce re nd lmin propri thickness of duodenum, jejunum nd ileum in the 4 groups. As it is noticele in Figure 1, Aqulend group hd the highest surfce re compred to control (P < 0.05), ut the lmin propri thickness ws not significntly different mong groups (P > 0.05) except cont-stress group which ws the lest (P < 0.05). On the other hnd, jejunum villi morphology results were different mong groups nd there were not ny sttisticlly significnt differences mong groups, lthough Aqulend group showed slightly thicker lmin propri compred to control (P > 0.05) nd oth of them were significntly thicker thn the 2 cold-hypoxic groups (P < 0.05) (Figure 2). Menwhile, ileum surfce re of control group ws incresed more thn other groups. However, the increment ws not significnt in comprison to Aqulend nd qu-stress groups (P > 0.05), ut similr to jejunum lmin propri, ileum lmin propri for Aqulend group ws slightly higher thn others. RV: TV rtio ws highest in cont-stress group nd lowest in oth stndrd temperture groups. Aqu-stress group gined lesser mount of RV: TV rtio (0.25) thn cont-stress group (0.28) (P > 0.05). In ddition, H: L rtio ws higher in cont-stress group (1.22) thn tht in qustress group (1.2) (P > 0.05) nd the differences etween Aqulend nd cont-stress groups were significnt Tle 2. Weekly Feed Conversion Rtio in 4 Groups t Dy 35 Aqulend Control Aqu-stress Cont-stress Week ± ± Week ± ± Week ± ± ± ± 0.24 Week ± ± ± ± C Week ± ± ± ± 0.374,,c Different letters within row demonstrte sttisticlly significnt differences (P < 0.05). Tle 3. Percent of Reltive Weight of Crcss, Liver, Spleen, nd Lung to Live Weight t Dy 35 in 4 Groups Crcss (%) Liver (%) Spleen (%) Lung (%) Aqulend ± ± ± ± Control ± ± ± ± Aqu-stress ± ± ± ± Cont-stress ± ± ± ± 0.030,,c Different letters within row demonstrte sttisticlly significnt differences (P<0.05). Tle 4. RV: TV nd H: L Rtios in 4 Groups t Dy 35 Aqulend Control Aqu-stress Cont-stress RV: TV 0.24 ± ± ± ± H: L 1.12 ± ± c 1.20 ± c 1.22 ± 0.016,,c Mens within row with the sme letter re not significntly different (P<0.05). Interntionl Journl of Enteric Pthogens Volume 5, Issue 2, My

4 (P < 0.05) (Tle 4). Duodenum mucos morphology c Discussion The primry role of proiotics in the first dys of irds lives is to inhiit the pthogens from occupying inner intestinl surfce s competitive exclusion which cn villus surfce re lmin propri Aqulend control Aqu-stress cont-stress Figure 1. Duodenum Villus Surfce Are nd Lmin Propri Thickness in 4 Groups t Dy 35.,,c Different letters within row demonstrte sttisticlly significnt differences (P < 0.05) Figure 2. Jejunum Villus Surfce Are nd Lmin Propri Thickness in 4 Groups t Dy 35,,c Different letters within row demonstrte sttisticlly significnt differences (P<0.05) Aqulend control Aqu-stress cont-stress Jejunum mucos morphology c c villus surfce lmin propri Figure 3. Ileum Villus Surfce Are nd Lmin Propri Thickness in 4 Groups t Dy 35.,,c Different letters within row demonstrte sttisticlly significnt differences (P<0.05). Ileum mucos morphology d villus surfce lmin propri Aqulend control Aqu-stress cont-stress fcilitte growth nd hold considerle promise for the helth cre industry. Growth performnce results in this study reveled tht ddition of Aqulend Avin proiotic to drinking wter during the first 3 dys of life could pprently decrese feed: gin rtio in comprison with control group t the end of the experiment (P < 0.05). On the other hnd, Aqulend decresed FCR in quils which were rised under cold-hypoxic stress sitution nd the decrement ws enough to e sttisticlly significnt in fourth week compred to cont-stress group nd lso to help this group not to hve ny significnt difference with control group. Effects of eneficil cteri on BW gin nd FCR hs een descried to e controversil. There re some results in greement with our dt which showed dvntgeous effects of proiotics nd syniotics on BW nd FCR On the other hnd reports suggesting no impct of proiotics on BW re lso ville. 19,20 This conflict might hve connection with dosge, concentrtion, strin nd form of cteri used nd lso the methods of using proiotics. 21 Bsed on our dt lthough not significnt, Aqulend group hd slightly higher reltive weights of crcss nd spleen. Furthermore, qu-stress group ws le to increse the percent of crcss, spleen nd lung in comprison with cont-stress group, lthough not significnt. Severl studies hve shown tht syniotics, preiotics nd proiotics hd no significnt positive effects on crcss yield of quils nd roilers. 19,22-25 There hve een such controversil results in this study regrding intestinl villi morphology. The villi surfces were igger in duodenum segments of Aqulend fed quils. Solis de los Sntos et l 26 reported tht the lrgest increses of villi ws in duodenum villi which ws significntly elevted in preiotic-treted irds. The jejunum nd ileum surfce re of cont-stress quils were reduced significntly compred to qu-stress irds (P < 0.05), which ws lso in greement with the study worked y Solis de los Sntos et l. 26 In ddition, lmin propri thickness of ileum nd jejunum ws elevted in Aqulend nd qu-stress groups compred to control nd cont-stress groups, respectively. Lmin propri thickness demonstrtes intestinl helth ecuse it consists of dendritic cells tht survey the contents of the lumen nd immunize the ird ginst infection y stimulting the dptive immune response, modifying mucin production nd defensin secretion, incresing gut motility, nd IgA production. 27 Thin scttered lmin propri is more quickly penetrted y pthogens; so it is n essentil component of the immune system. 28 Results regrding right hert filure index shows tht RV/TV rtio ws higher in cont-stress group versus qu-stress group which might show the inhiition of right ventriculr filure nd scites syndrome in qustress group. However, signs of scites syndrome were not oservle in quils s significnt hydropericrdium nd mortlity. Although occurrence of scites syndrome is more common in roilers thn in other poultry Interntionl Journl of Enteric Pthogens Volume 5, Issue 2, My 2017

5 Sffr nd Khjli 29 discussed tht the lower numer of mortlity in proiotic-receiving groups due to scites syndrome might correlte with decresed mmoni production in the intestine. Our dt is lso in greement with those of Solis de los Sntos et l 26 tht declred the first connection of gut development to scites syndrome in roiler chicks. Dt out H: L rtio ws lso represented to e the highest in cont-stress nd the lowest in Aqulend group tht shows lesser stress in Aqulend group under norml temperture. It is suggested tht proiotic could llevite the stress of feed deprivtion nd enhnce the immune-competence of chickens 30 which is in greement with the dt presented here. Both norml environment groups gined fewer H: L rtio thn two cold hypoxic environment groups. Moreover, cont-stress group showed slightly higher mount of H: L rtio which might revel higher stress induced y cold-hypoxic sitution thn qu-stress group. Totlly oth Aqulend groups showed etter response to physiologicl responses. Conclusion This study showed tht ddition of Aqulend Avin proiotic in the first 3 dys of life cn significntly reduce FCR, either in norml, or in cold-stressful situtions. This report is the first one to induce right hert filure in Jpnese quil y cold hypoxic sitution to evlute the effects of inclusion of proiotic on reducing RV: TV rtio. Treted irds were lso more resistnt to stressful conditions while compring H: L rtio. Dt regrding to intestinl mucos morphology ws controversil ut Aqulend Avin proiotic ws le to elevte duodenum villi surfce nd lso jejunum nd ileum lmin propri thickness. Authors Contriutions Study concept nd design: AKZM; Acquisition of dt: MHMH, SST, HP nd AH; Anlysis nd interprettion of dt: AKZM nd MHMH; Drfting of the mnuscript: AKZM nd MHMH. Ethicl Approvl The Institutionl Animl Cre nd Use Committee of Shhrekord University pproved the study. Conflict of Interest Disclosures The uthors declre tht they hve no confict of interests. Finncil Support Vice Chncellor for Reserch t Shrekord University supported the study. Acknowledgements The uthors re gretly indeted to Vice Chncellor for Reserch t Shrekord University to the funds grnted for this work. References 1. Hosseini F, Mehrei Hmzekolei MH, Zmni Moghddm AK, Arh H, Tohidifr SS. Norml ultrsonogrphic imges of reproductive orgns of femle Jpnese quils (Coturnix coturnix jponic): lortory niml model. L Anim. 2017;51(1): doi: / El Trny M. Impct of cge stocking density on egg lying chrcteristics nd relted stress nd immunity prmeters of Jpnese quils in sutropics. J Anim Physiol Anim Nutr (Berl). 2016;100(5): doi: /jpn Nguyen P, Greene E, Donoghue A, Huff G, Clrk F, Dridi S. A new insight into cold stress in poultry production. Adv Food Technol Nutr Sci Open J. 2016;2(1): Smnt G, Ghosh C, Smnt G. Sfe food fro, roiler chicks nd jpnese quil with lterntive ntiiotic growth promoters. Interntionl Journl of Bio-Resource, Environment nd Agriculturl Sciences. 2016;2(1): Swyne DE, Glisson JR, McDougld LR, Noln LK, Surez DL, Nir VL. Diseses of Poultry. John Wiley & Sons; Vuong CN, Chou W-K, Hrgis BM, Berghmn LR, Bielke LR. Role of proiotics on immune function nd their reltionship to ntiiotic groewth promoters in poultry: rief review. Int J Proiotics Preiotics. 2016;11(1). 7. Sd N, Delttre C, Urdci M, Schmitter J-M, Bressollier P. An overview of the lst dvnces in proiotic nd preiotic field. LWT Food Sci Technol. 2013;50(1):1-16. doi: /j. lwt Frei R, Akdis M, O Mhony L. Preiotics, proiotics, syniotics, nd the immune system: experimentl dt nd clinicl evidence. Curr Opin Gstroenterol. 2015;31(2): Reid G. Proiotics: definition, scope nd mechnisms of ction. Best Prct Res Clin Gstroentero. 2016;30(1): doi: /j.pg Zhng Z, Kim I. Effects of multistrin proiotics on growth performnce, pprent ilel nutrient digestiility, lood chrcteristics, cecl microil shedding, nd excret odor contents in roilers. Poult Sci. 2014;93(2): Huff G, Huff W, Rth N, El-Gohry F, Zhou Z, Shini S. Efficcy of novel preiotic nd commercil proiotic in reducing mortlity nd production losses due to cold stress nd Escherichi coli chllenge of roiler chicks. Poult Sci. 2015;94(5): doi: /ps/pev Tlzdeh F, Myhi M, Zeinli S. Effect of Aqulend Avin proiotic strins of Lctocillus, Streptococcus nd Bifidocterium on systemic ntiody responses ginst influenz nd Newcstle diseses vccine in roiler chickens. Int J Enteric Pthog. 2016;4(2):e doi: / ijep Lucs AM, Jmroz C. Atls of Avin Hemtology. Wshington, DC: U.S. Dept. of Agriculture; Skmoto K, Hirose H, Onizuk A, et l. Quntittive study of chnges in intestinl morphology nd mucus gel on totl prenterl nutrition in rts. J Surg Res. 2000;94(2): Awd W, Ghree K, Adel-Rheem S, Böhm J. Effects of dietry inclusion of proiotic nd syniotic on growth performnce, orgn weights, nd intestinl histomorphology of roiler chickens. Poult Sci. 2009;88(1): Mountzouris K, Tsirtsikos P, Klmr E, Nitsch S, Schtzmyr G, Fegeros K. Evlution of the efficcy of proiotic contining Lctocillus, Bifidocterium, Enterococcus, nd Pediococcus strins in promoting roiler performnce nd modulting cecl microflor composition nd metolic ctivities. Poult Sci. 2007;86(2): Smli HE, Senkoylu N, Koc F, Knter M, Agm A. Effects of Enterococcus fecium nd dried whey on roiler performnce, gut histomorphology nd intestinl microiot. Arch Anim Nutr. 2007;61(1): doi: / Agool A, Omidiwur B, Odu O, Odupitn F, Iyyi E. Effect of proiotic nd toxin inder on performnce, intestinl microiot nd gut morphology in roiler chickens. J Anim Sci Adv. 2015;5(7): Shrifi MR, Shms-Shrgh M, Dstr B, Hssni S. The effect of dietry protein levels nd syniotic on performnce prmeters, lood chrcteristics nd crcss yields of Jpnese quil (Coturnix coturnix Jponic). Itl J Anim Sci. Interntionl Journl of Enteric Pthogens Volume 5, Issue 2, My

6 2011;10(1):e Srngi NR, Bu L, Kumr A, Prdhn C, Pti P, Mishr J. Effect of dietry supplementtion of preiotic, proiotic, nd syniotic on growth performnce nd crcss chrcteristics of roiler chickens. Vet World. 2016;9(3): doi: /vetworld Klvthy R, Adullh N, Jlludin S, Ho Y. Effects of Lctocillus cultures on growth performnce, dominl ft deposition, serum lipids nd weight of orgns of roiler chickens. Br Poult Sci. 2003;44(1): Ckir S, Midilli M, Erol H, et l. Use of comined proioticpreiotic, orgnic cid nd vilmycin in diets of Jpnese quils. Rev Med Vet (Toulouse). 2008;159(11): Chumpwdee S, Chinrsri O, Sntweesuk S. Effect of dietry inclusion of cssv yest s proiotic source on growth performnce nd crcss percentge in Jpnese quils. Pkistn J Nutr. 2009;8(7): Pelíci K, Mendes A, Sldnh E, et l. Proiotic nd preiotic utiliztion in diets for free-rnge roiler chickens. Revist Brsileir de Ciênci Avícol. 2004;6(2): doi: / S X Shin T, Ky I, Unl Y, Elmli DA. Dietry supplementtion of proiotic nd preiotic comintion (Comiotics) on performnce, crcss qulity nd lood prmeters in growing quils. J Anim Vet Adv. 2008;7(11): Solis de los Sntos F, Frnell M, Tellez G, et l. Effect of preiotic on gut development nd scites incidence of roilers rered in hypoxic environment. Poult Sci. 2005;84(7): Mcpherson AJ, Hrris NL. Interctions etween commensl intestinl cteri nd the immune system. Nt Rev Immunol. 2004;4(6): Grtner LP, Hitt JL. Colour Textook of Histology. Phildelphi: WB Sunders; Sffr A, Khjli F. Appliction of mel feeding nd skip--dy feeding with or without proiotics for roiler chickens grown t high-ltitude to prevent scites mortlity. Am J Anim Vet Sci. 2010;5: Yurong Y, Ruiping S, Shimin Z, Yio J. Effect of proiotics on intestinl mucosl immunity nd ultrstructure of cecl tonsils of chickens. Arch Anim Nutr. 2005;59(4): doi: / Interntionl Journl of Enteric Pthogens Volume 5, Issue 2, My 2017