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S. M. Weedman, M. H. Rostagno, J. A. Patterson, I. Yoon, G. Fitzner and S. D. Eicherintestinal microbial ecology of weanling pigs

Yeast culture supplement during nursing and transport affects immunity and

doi: 10.2527/jas.2009-25392011, 89:1908-1921.J ANIM SCI

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at Serials/Acq. Dept., Library on June 21, 2011jas.fass.orgDownloaded from
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Yeast culture supplement during nursing and transport affectsimmunity and intestinal microbial ecology of weanling pigs1,2

S. M. Weedman,* M. H. Rostagno, J. A. Patterson,* I. Yoon, G. Fitzner,and S. D. Eicher3

*Purue University, West Lafayette, IN 47907; Livestock Behavior Research Unit, USDA-ARS,

West Lafayette, IN 47907; an Diamon V Mis Inc., Cear Rapis, IA 52407-4570

ABSTRACT: The objectives of this stuy were to e-termine the infuence of a Saccharomyces cerevisiaefer-mentation prouct on innate immunity an intestinamicrobia ecoogy after weaning an transport stress.In a ranomize compete bock esign, before weaningan in a spit-pot anaysis of a 2 2 factoria arrange-ment of yeast cuture (YY) an transport (TT) afterweaning, 3--o pigs (n = 108) were ranomy assignewithin itter (bock) to either a contro (NY, mik ony)

or yeast cuture iet (YY; eivere in mik to provie0.1 g of yeast cuture prouct/kg of BW) from 4 to 21.At weaning ( 21), ranomy, one-haf of the NY anYY pigs were assigne to a 6-h transport (NY-TT anYY-TT) before being move to nursery housing, anthe other one-haf were move irecty to nursery hous-ing (NY-NT an YY-NT, where NT is no transport).The yeast treatment was a 0.2% S. cerevisiaefermen-tation prouct an the contro treatment was a 0.2%grain bank in fee for 2 wk. On 1 before transportan on 1, 4, 7, an 14 after transport, boo was co-ecte for eukocyte assays, an mesenteric ymph noe,jejuna, an iea tissue, an jejuna, iea, an cecacontents were coecte for To-ike receptor expression(TLR); enumeration of Escherichia coli, tota coiforms,an actobacii; etection of Salmonella; an microbia

anaysis. After weaning, a yeast transport interactionfor ADG was seen (P= 0.05). Transport affecte (P=0.09) ADFI after weaning. Yeast treatment ecreasehematocrit (P= 0.04). A yeast transport interactionwas foun for counts of white boo ces (P= 0.01)an neutrophis (P= 0.02) an for the neutrophi-to-ymphocyte ratio (P= 0.02). Monocyte counts reveaea transport (P= 0.01) effect. Interactions of yeast transport (P= 0.001) an yeast transport ay (P

= 0.09) for TLR2 an yeast transport (P= 0.08) forTLR4 expression in the mesenteric ymph noe wereetecte. Day affecte actobacii, tota coiform, anE. colicounts. More pigs were positive for Salmonellaon 7 an 14 than on 4, an more YY-TT pigs werepositive (P= 0.07) on 4. The number of bans formicrobia ampicons in the ieum was greater for pigsin the contro treatment than in the yeast treatment on 0, an this number tene to ecrease (P= 0.066)between 1 an 14 for a pigs. Simiarity coefficientsfor jejuna contents were greater (P= 0.03) for pigs feNY than for those fe YY, but pigs fe YY ha greatersimiarity coefficients for iea (P= 0.001) an ceca (P= 0.058) contents. The number of yeast transport ay interactions emonstrates the compexity of thestress an ietary reationship.

Key words: immune response, intestina microbiota, pig, transport, weaning, yeast cuture

2011 American Society of Animal Science. All rights reserved. J. Anim. Sci. 2011. 89:19081921oi:10.2527/jas.2009-2539

INTRODUCTION

After weaning, piget immunity is compromise by

exposure to mutipe stressors. These stressors ea

to reuce fee intake an ater the eveopment ancommunity structure of the commensa bacteria pop-uation in the intestine. Consequenty, weaning an

transport stress enhances the vunerabiity to cooniza-tion by pathogenic bacteria. Thus, eveoping methosto stimuate the innate immune system of the pig mayecrease susceptibiity of the host.

Severa attempts have been mae to aeviate weaningstress by feeing ietary mouators, incuing spray-rie pasma (Pierce et a., 2005), vitamin E (Fragouet a., 2006), atere CP an fermentabe carbohyrates(Bikker et a., 2006), nonigestibe oigosaccharies annonstarch poysaccharies (Van Neve et a., 2005),

1Mention of trae names or commercia proucts in this pubica-tion is soey for the purpose of proviing specific information anoes not impy recommenation or enorsement by the USDA.

2Fune in part by Diamon V Mis Inc. (Cear Rapis, IA).3Corresponing author: [email protected] September 30, 2009.Accepte January 11, 2011.

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an yeasts (Li et a., 2006) an yeast proucts (Li eta., 2005; Eicher et a., 2006; Hahn et a., 2006). Yeastsuppements are promising because they are capabeof enhancing periphera as we as intestina immunity.Yeast cuture is a rie fermente prouct composeof resiua ea yeast (Saccharomyces cerevisiae) anthe meia on which it was grown. Portions of the yeastce wa, incuing -gucans an mannans, have im-

munomouating properties. The -gucan componentcan mouate both the innate an aaptive arms ofthe immune system (Xiao et a., 2004). Stuies of theeffects of yeast cuture or yeast extracts on immunityin mice are abunant, but few pubications have a-resse the effects of yeast on immune mouation inswine. Even fewer stuies have incorporate the effectsof yeast extracts or yeast cutures on commensa bac-teria. The objectives of this stuy were to eterminethe infuence of a S. cerevisiaefermentation prouct oninnate immunity an intestina microbia ecoogy afterweaning an transport stressors in pigs.

MATERIALS AND METHODS

Anima use was approve by the Purue AnimaCare an Use Committee.

Animals, Experimental Design, and Housing

The experiment was conucte at the Purue Uni-versity Swine Unit in 2 repications, 1 in the fa an1 in the spring. One hunre eight pigets (54 perrepication) from 24 mutiparous sows (12 per repica-tion) were iniviuay weighe at 72 h after farrowing.Within each itter, 4 pigets were chosen base on simi-

ar BW, an BW an sex were baance among ittersan pigets were assigne to 1 of 4 treatments arrangein a factoria array. Factors were yeast suppementation[yeast (YY) or no yeast (NY)] an transport [trans-porte (TT) an no transport (NT)]. The experimentwas anayze as 2 separate experiments, preweaningan postweaning. The preweaning ata was treate asa ranomize compete bock esign, with yeast treat-ment an the contro iet as treatments an itter asthe bocking factor. The postweaning ata were ana-yze as a spit pot in time with a 2 2 factoria ar-rangement of the treatments yeast cuture (YY) antransport (TT). At experimenta 0, the yeast suppe-ment (XPC, Diamon V Yeast Cuture, Diamon VMis Inc., Cear Rapis, IA) was eivere in mik toprovie 0.1 g of yeast cuture prouct/kg of BW from 4 to 21. At weaning ( 21), one-haf of the NY an YYpigs (ranomy assigne within itter) were transportefor 6 h (NY-TT an YY-TT) before being move tonursery housing, an the other one-haf were moveirecty to nursery housing (NY-NT an YY-NT). Pigsin the YY-NT an YY-TT treatments were suppe-mente with 0.2% yeast suppement an those in the

NY-NT an NY-TT treatments receive a 0.2% grainbank in fee for 2 wk.

The yeast cuture was fe in warme (37C) whoebovine mik at 0.1 g of yeast cuture/kg of BW. Yeastcuture [(0.1 g of yeast cuture/kg of mean BW of thepigs) 54] was mixe into 54 mL of mik (1 mL/pig-et). Iniviua voumes of mik containing yeast cuturewere aministere oray to each piget to achieve 0.1

g of yeast cuture/kg of BW. Pigs in the contro treat-ments receive ony mik ajuste for BW. IniviuaBW of pigets were recore twice weeky, an yeastcuture in the mik aiquots was ajuste unti wean-ing at approximatey 21 of age. One aitiona pigetfrom each of 12 itters at each repication was ranomyassigne to the nontransporte treatments (NY-NT orYY-NT) as the baseine contros before transport. Pig-ets were coor coe with paint markers for treatmentientification.

At weaning, 48 pigets (24 per repication), 1 NY an1 YY ranomy assigne from each itter, were trans-porte (TT) for 6 h by traier (bee with shavings

with ampe space aowance, 0.18 m2

/piget; Sutheranet a., 2009) over a esignate route to incue high-way an interstate roas (2 transports). Pigets werepenne within the transport vehice with ittermatesbecause it is we estabishe that isoation is an ex-treme stressor (Herskin an Jensen, 2000; Kanitz et a.,2004). Pigets reste an i not fight at any point inthe transport, presumaby because they were with it-termates an ominance was areay estabishe, anbecause they were not in competition for foo or wa-ter. The remaining pigets (NT) were move into thenursery environment at the same time the transportepigets were oae onto the traier.

Pigets were house iniviuay after weaning. Penswere arrange by ay of euthanasia within the room,with treatments ranomy pace within the ay of eu-thanasia pens. This pacement was teste as a ranomvariabe, an pacement within the room was foun notto be significant (P> 0.10). Each treatment was repre-sente in an en pen at east once. Experimenta ietswere compose of phase 1 an phase 2 iets (Tabe 1)with a grain bank (grain mixture that was use tomake the yeast cuture) to repace the yeast cuture.For the yeast treatments, yeast cuture was incue at0.2% of the nursery iet an the iet was ajuste forthe 15% CP suppie by the yeast prouct. Weaningpigs were fe free choice for up to 2 wk after weaning.On the ay of weaning before transport, 6 baseine pig-ets from each of the ietary treatments (NY or YY)were humaney kie with 0.22 mL of BeuthanasiaD(Schering-Pough Anima Heath Corporation, Summit,NJ) per kiogram of BW, foowe by exsanguination.These serve as en points ( 21) for the preweaningpigs an as the baseine vaues for postweaning pigs.Immeiatey after the pigets returne from transportan at 4, 7, an 14 after transport, 6 pigets from

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each treatment group (NY-NT, YY-NT, NY-TT, anYY-NT) were kie for each repication.

Sample Collection and Measurements

Performance. After birth, iniviua BW of pig-ets were recore twice weeky unti weaning. Pigetswere weighe at weaning an on 4, 7, or 14 after

transport. Iniviua fee consumption was recore ateach weighing interva after weaning an fee conver-sion was etermine.

Hematology. Before euthanasia, pigets were re-straine using a V-trough, an juguar boo was co-ecte into 2 vacuum tubes: one 5-mL EDTA tube (12mg in a 7-mL raw tube; Becton Dickinson, FrankinLakes, NJ) an one 10-mL tube containing aci citrateextrose [ACD; 1.5 mL of soution A (22.0 g of trisoi-um citrate; 8 g of citric aci; an 24.5 g of extrose/L)in an 8.5-mL raw tube; Becton Dickinson]. Boo wasstore on ice unti assaye. Boo from the EDTA tubeswas use for a 5-part ifferentia hematoogy anaysis

using a Hemavet 950 Hematoogy Anayzer (Drew Sci-entific, Daas, TX). Boo ce measures consiste ofhematocrit percentage; counts of tota white boo ces(WBC), neutrophis, monocytes, an ymphocytes;an percentages of neutrophis, monocytes, an ym-phocytes. Boo coecte into vacuum tubes containingACD was use for periphera eukocyte fow cytometricanaysis.

Leukocyte Function. Custer of ifferentiation(CD) 14 is part of the ipopoysaccharie receptor thatis primariy on monocytes, macrophages, an enriticces invove in antigen processing an presentation.The expression of CD18, as part of an ahesion mo-ecue, is upreguate on activate ces, particuary onneutrophis. Ce surface expression of CD14 an CD18was anayze using 3 aiquots of 500 L each of whoeboo from the ACD tubes. Boo was incubate ina 37C water bath for 30 min before aition of anti-boies. To the first tube, 10 L of phycoerythrin-con-jugate, monocona mouse anti-human CD14, coneTUK4 (DakoCytomation, Carpinteria, CA) an 20 Lof monocona mouse anti-human eukocyte functionaantigen-1, -chain/fuorescein isothiocyanate CD18,cone 6.7 antiboies (BDPharmingen, San Jose, CA;100 mg/L) were ae to etect ce surface marker

expression. FuoSpheres, carboxyate-moifie micro-spheres (12.5 L; 1.0 m; Moecuar Probes, Eugene,OR), were ae to the secon tube for microbeaphagocytosis anaysis. Another tube remaine withoutantiboy to serve as unstaine contro ces an aowegating of autofuorescence. Sampes were incubate ina 37C water bath for 30 min. Ces were yse by hy-potonic ysis, in which 900 L of co sterie water wasae to each sampe tube for 45 s. After re booce ysis, 0.1 mL of 10 Hanks Baance Sat Sou-tion (HBSS) was ae to restore isotonicity. Sampeswere centrifuge for 3 min at 1,800 gat 4C an ce

ysis was repeate. Sampes were washe with 1 mLof 1 HBSS an centrifuge at 1,800 gfor 3 min at4C. To preserve sampes for fow cytometric anaysis,ces were suspene in 500 L of 2% paraformaehyein 1 HBSS. Fow cytometry was conucte using aCouter Epics XL-MCL Fow Cytometer an SystemII software (Beckman Couter Inc., Miami, FL). Thefow cytometer use a 488-nm air-cooe argon aser forexcitation, a 525-ban pass fiter for etection of fuo-rescein isothiocyanate emissions, an a 575-ban passfiter for etection of phycoerythrin emission. For eachsampe, a tota popuation of 10,000 ces was anayze.

RNA Extraction and PCR. Mesenteric ymphnoes (MLN) ocate proximay to the ieoceca junc-tion, an one 3-cm section of the ieum an the jejunumwere harveste asepticay from each piget after exsan-

guination. Sections were strippe of the serosa ayeran cut aong the mesentery to expose the inner area ofthe tissue. Tissue was rinse with sterie 1 HBSS anpace in 3.6-mL cryovia tubes with 3 mL of RNAater(Ambion Inc., Austin, TX) for RNA extraction anstore at 80C. Sampes were homogenize an RNAwas extracte by using a tissue griner an RNeasyMini Kit (Qiagen, Vaencia, CA). Extracte RNA wasquantifie using the GeneQuant pro RNA/DNA Cacu-ator (Amersham Biosciences Corp., Piscataway, NJ).

Compementary DNA was ampifie with mastermix, compose of reagents incue in the TaqMan

Table 1. Ingreient composition of experimenta iets(as-fe basis) fe to pigs after weaning ( 21 to 35) thatincue the yeast suppement or a grain bank

Ingreient, % Phase 1 Phase 2

Finey groun corn 34.8 39.3Spray-rie whey 25.0 25.0Soybean mea, 47.5% 12.5 20.0Anima pasma 6.7 5.0

White swine grease 6.0 2.5Fishmea, menhaen 6.0 2.5Lactose 5.0 Boo mea, swine 1.7 2.5Dicacium phosphate 0.5 1.1Cacium carbonate 0.4 0.6Zinc oxie, 72% 0.375 0.375Sat pain (Keer Mixing1) 0.25 0.30Suppement2 0.20 0.20dl-Methionine 0.15 0.15l-Lysine hyrochorie 0.15 0.15Vitamin premix3 0.15 0.15Nonsufur trace minera premix4 0.125 0.125Seenium, 0.05% 0.05 0.05

1Morton Keer Mixing Sat (Morton Sat, Chicago, IL).2Grain bank or Saccharomyces cerevisiaeyeast suppement (XPC,

Diamon V Mis Inc., Cear Rapis, IA). The yeast suppement pro-vies not ess than 15% CP, 1.50% fat, an 22.0% fiber.

3Provie the foowing per kiogram of iet (as-fe basis): vitaminA, 6,600 IU; vitamin D3, 660 IU; vitamin E, 44.0 IU; menaione, 2.2mg; vitamin B12, 38.5 g; ribofavin, 8.8 mg; d-pantothenic aci, 22.0mg; niacin, 33.0 mg.

4Provie the foowing per kiogram of iet (as-fe basis): Fe, 84.88mg; Zn, 84.88 mg; Mn, 10.5 mg; Cu, 7.88 mg; I, 0.29 mg; Se, 0.3 mg.

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Reverse Transcription Reagent Pack (Appie Biosys-tems, Foster City, CA) an prepare in a sterie ispos-abe reservoir (Vistaab Technoogies, Mt. Kisco, NY),for a 100-L fina voume. It containe 50 units/Lof Muti-Scribe reverse transcriptase, 25 mM MgC2,2.5 Mranom hexamers, 0.4 units/L of ribonuceaseinhibitor, 50 Meoxynuceotie 5-triphosphate, anTaqMan reverse-transcription buffer (TaqMan reverse-

transcription reagents, Appie Biosystems). SampeRNA an ribonucease-free water (Ambion Inc.) wereae to the master mix at appropriate quantities.Compementary DNA was ampifie (Hybai PCRExpress Thermo Cycer, Miwest Scientific, St. Louis,MO) using the foowing cycing conitions: 50C for 2min of activation, foowe by 40 cyces of 95C for 10min, 95C for 15 s, 60C for 1 min, an a fina stage of60C for 5 min, with a hoing temperature of 4C. Theresuting cDNA was store at 80C unti PCR wasperforme.

After reverse transcription, the reative abunanceof To-ike receptor (TLR) 2 an TLR4 were eter-

mine by quantitative reverse-transcription PCR usingprimers an probes (Tabe 2) eveope using PrimerExpress an synthesize by Appie Biosystems. Theabunance of genes of interest reative to the quantityof 18S rRNA in tota RNA isoate from homogenizeMLN, jejuna, an iea tissue was quantifie using anABI Prism 7000 Sequence Detection System (AppieBiosystems). Reactions were carrie out using the ap-propriate TaqMan probe (200 nM), forwar primeran reverse primer, PCR master mix (Appie Biosys-tems), an 5 L of the cDNA sampe an a programconsisting of an initia step at 50C for 2 min, fooweby 50 cyces of 10 min at 95C, 20 s at 95C, an 1 min

at 60C. Commerciay avaiabe eukaryotic 18S rRNAprimers an probes (Appie Biosystems) were use asan enogenous contro.

Microbial Analysis. Intestina contents (2 mL)were coecte asepticay after saughter from the je-junum, ieum, cecum, an MLN for microbia anaysis.Iniviua contents of the jejunum, ieum, an cecumwere ivie into 2 aiquots, one for microbia profiingusing enaturing graient-ge eectrophoresis (DGGE)an one for enumeration of Escherichia coli, tota co-

iforms, an Lactobacillus. Part of the secon aiquotan MLN were seectivey enriche for isoation of Sal-monella.

To etect E. coli, tota coiforms, an Lactobacillus,intestina contents were seriay iute in buffere pep-tone water (Neogen, Lansing, MI) an incubate over-night at 37C. Sampes (1 mL) were pate on Petrifim(3M, St. Pau, MN) for enumeration of E. colian tota

coiforms an on Rogosa agar (Difco, Lawrence, KS) forenumeration of actobacii an were incubate for 24 hat 37C. To etect Salmonella, ceca contents an MLNwere iute 10-fo in tetrathionate broth (Neogen)an incubate overnight at 37C. Sampes (1 mL) weretransferre to 10 mL of Rappaport-Vassiiais broth(Neogen) an incubate for 24 h at 42C. Loops ofRappaport-Vassiiais enrichment were streake ontoXLT4 pates (Neogen) an incubate for 24 h at 37C.Back-coore coonies were ientifie as Salmonellapositive by using Rambach agar (DRG InternationaInc., Mountainsie, NJ).

For microbia profiing, jejuna, iea, an ceca con-

tents were coecte into 1.5-mL microfuge tubes anfrozen at 80C unti assaye. Tissues were homoge-nize (UtraTurrax T 25, IKA Works Inc., Wiming-ton, NC), an genomic DNA was extracte from bothtissues an contents by using a Fast DNA Spin Kitfor Soi (MP Biomeicas LLC, Soon, OH). ExtracteDNA was quantifie using the GeneQuant pro RNA/DNA Cacuator (Amersham Biosciences Corp.) anthen iute for PCR anaysis.

Poymerase chain reaction proceures were as e-scribe by Wiiams et a. (2008). Briefy, the V3 re-gion of the 16S rDNA was ampifie an bans wereseparate on 30 to 60% enaturing graient poyacry-

amie ges (8%) using a DCoe Universa MutationDetection System (Bio-Ra, Hercues, CA). Sampeswere eectrophorese at 60C for 5.5 h. Fragment pat-tern reateness was etermine using BionumericsSoftware (version 2.5, Appie Maths, Austin TX).Simiarity coefficients were erive from the pairwisecomparison of baning patterns between 2 sampesan were cacuate using the DICE agorithm forbans within sampe, as escribe by Burkhoer eta. (2008).

Table 2. Primers an probes use for quantitative reverse-transcription PCR for To-ike receptor (TLR) 2 anTLR4 gene expression

Gene Primer or probe Sequence Accession number

TLR2 Forwar primer (sense) TGT GGC CAT CGC TGC TAA C AY392087Reverse primer (antisense) GGG ACA CCA CGA CAA TAA CCT TProbe VIC1 TCA TCC AGG AAG GTT TCC ACA AAA GTC G MGBNFQ2

TLR4 Forwar primer (sense) TGT GGC CAT CGC TGC TAA C AB232527Reverse primer (antisense) GGG ACA CCA CGA CAA TAA CCT TProbe VIC TCA TCC AGG AAG GTT TCC ACA AAA GTC G MGBNFQ

1VIC is a proprietary esignation (Appie Biosystems, Foster City, CA).2MGBNFQ = minor groove biner nonfuorescent quencher.



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Statistical Analysis

Data were teste for normaity using the Univariateproceure (SAS Inst. Inc., Cary, NC) an transformeas necessary: WBC were square root transforme,a microbia anaysis were natura og-transforme,an iea TLR2 an TLR4 an jejuna TLR2 wereog-transforme. Data are presente as east squaresmeans SE. The treatments were anayze as a facto-ria arrangement of yeast an transport. Performance(BW, ADG, an fee conversion) was anayze as apreweaning segment an as a postweaning segment.Boy weight, ADG, ADFI, an G:F were anayze asrepeate measures using mixe moe proceures (SASInst. Inc.), with pig as the experimenta unit. For thefirst 21- anaysis, the fixe effects, consisting of the 2ietary treatments (yeast an contro), ay, an theirinteractions, were teste as a repeate measure. For thefirst 21 , 0 BW was use as a covariate. The firstcovariate was use because BW ifferences betweentreatments were etecte at experimenta 0.

For postweaning ata, the mixe moes proceuresof SAS were use to anayze the spit-pot esign, withthe factoria arrangement of yeast an transport as thewhoe pots an ay as the subpot. The fixe effectsteste were ay an treatments (yeast, transport, antheir interaction). For the ast 2 wk when the yeastwas eivere in ry fee, BW at 21 was use as acovariate. The secon covariate was use because ofanticipate ifferences cause by the treatments uringthe nursery phase of the experiment. Season (fa anspring) an room were teste as ranom variabes anremove because they were not significant. Error termswere etermine by the mixe moes proceures, which

eiminates the nee for error term construction by au-tomaticay incorporating the correct error term intotest statistics. Repication an sex were teste as ran-om variabes for each an were incue in the moeony when they were significant. The same statisticaanayses were use for boo anaysis an eukocytefunctions (hematocrit percentages; WBC counts; neu-trophi, ymphocyte, an monocyte counts an percent-ages; neutrophi-to-ymphocyte ratio; CD14 an CD18ce surface expression; an phagocytic ces), micro-bia anaysis (E. coli, actobacii, an tota coiforms),an RNA expression of pathogen recognition moecues(TLR2 an TLR4). Treatment ifferences were eter-

mine using Fishers protecte LSD an Tukey-Kramermeans separation. Significance is reporte at = 0.10.However, exact probabiity vaues are provie for theiscretion of the reaer.

Salmonellaata were anayze by 2, an the effectsof repication, treatment, ay, an sex were teste.Chi-square anaysis was use to separate ay effectswithin treatments an treatment effects within ays forSalmonellacounts. The tota number of bans, withina sampe, an simiarity coefficients, within treatmentsan across treatments (cross-proucts), of DGGE ata

were statisticay anayze as ANOVA using the PROCMIXED comman of SAS as escribe by Burkhoeret a. (2008), with treatment an ay as the variabes.Cross-proucts were use as an estimate of no treat-ment effect for instances in which simiarity vaues werenumericay simiar but baning patterns were ifferent(Burkhoer et a., 2008).

RESULTS

Performance

Initia BW (Tabe 3) was greater for pigets in theYY than in the NY treatment (P= 0.02). Therefore, 0 BW was use as a BW covariate for the first 21 .Yeast was not significant (P= 0.11), but a ay effect(P= 0.001) was etecte in the first 21 . Boy weightat 21 was use as a covariate for the remaining 2 wkof growth anaysis. A yeast transport interaction (P= 0.05) an a ay effect (P= 0.001) were etecte inthe anaysis of ADG for 21 to 35 of the stuy. Aver-

age aiy fee intake (Tabe 3) was affecte (P= 0.09)by transport uring wk 3 (the first week after weaning).Fee efficiency was not affecte by treatments (Tabe3). Pigs suppemente with yeast ha esser hematocritpercentages than nonsuppemente pigs (P = 0.04),an after 1, hematocrit percentages increase for apigs (P= 0.001) through the en of the stuy.

Hematology

White boo ce counts (Tabe 4) were affecte by ayeast transport interaction (P= 0.01) an a yeast transport ay interaction (P= 0.04). On 1, WBC

counts were ess for the YY-TT treatment than for theNY-NT treatment. By 4, pigets in the NY-TT anYY-NT groups ha greater WBC counts than pigetsin the NY-NT or YY-TT group. A yeast transportinteraction (P = 0.02) an a ay effect (P = 0.01)were etecte for neutrophi counts (Tabe 4), such thatpigs in the YY-TT group ha smaer neutrophi countsthan i pigs in the NY-TT or YY-NT group, an neu-trophi numbers increase throughout the stuy. Lym-phocyte counts ha a ay effect (P= 0.01) an a yeast transport ay interaction (P= 0.02) such that on 4, pigs in the YY-TT group ha the east ympho-cytes compare with pigs in the NY-TT an YY-NT

group, but on 14, pigets in the YY-NT group hathe east ymphocyte numbers compare with pigets inthe NY-NT group. Monocyte counts reveae a trans-port effect (P = 0.01) an a ay effect (P = 0.01).Monocyte counts of transporte pigs were greater thanthose of nontransporte pigs, an counts increasethroughout the stuy foowing transport. Anaysis ofneutrophi-to-ymphocyte ratios (Tabe 4) reveae ayeast transport ay interaction (P= 0.04), a yeast transport interaction (P= 0.04), a ay effect (P=0.04), an a transport effect (P= 0.06). On 1, pig-

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Table

3.Performanceofpigletsfrombirthto35dofage1forpigsfedayeastcultureorthecarrieronlyfromd3to35andtransportedfor6hornot

transportedatweaning(d21)in

a22factorialarrangementoftreatmentsford21to35

Item

Treatmentmean2

SEM

P-value

NY-NT

NY-TT

YY-NT

YY-TT

Y

east

Day

Trn3

Yeast

day

Yeast

trn

Yeast

trnday

InitialBW,4d

3,kg

1.99

2.05

0.06

0.02

Wk1,4 d3to7

BWatd7,kg(ADG,kg/d)

2.72(0.23)

2.74(0.20)

0.16(0.02)

0.11

0.001

0.45

Wk2,4 d7to14

BWatd14,kg(ADG,kg/d)

4.61(0.30)

4.75(0.29)

0.16

Wk3,4 d14to21


6.43(0.26)

6.22

5.88(0.23)

6.02

0.26(0.02)

Postweaningwk4and5,5

d21to35


7.48(0.17)

7.05(0.12)

7.14(0.18)

7.82(0.26)

0.26(0.02)

0.84

0.001

0.77

0.05

0.21


8.69(0.25)

9.4(0.29)

9.58(0.29)

9.10(0.24)

0.26(0.04)

Wk3ADFI,kg/d,d21to28

0.23

0.22

0.23

0.19

0.02

0.18

0.09

0.24

ADFI,kg/d,d28to35

0.58

0.64

0.63

0.58

0.05

0.83

0.89

0.22

G:Fwk4,d21to28

0.45

0.46

0.47

0.43

0.05

0.37

0.16

0.68

G:Fwk5,d28to35

0.64

0.71

0.65

0.69

0.08

0.84

0.71

0.45

1Leastsquaresmeans;noneweretransformed.

2NY-NT=controldietandnottransp

orted;NY-TT=controldietandtransported;YY-NT=yeastculturediet(XPC,D

iamondVMillsInc.,CedarRapids,IA)a

ndnottransported;YY-TT

=yeastculturedietandtransported.

3Trn=transport.

4NY-NT(n=29),NY-TT(n=24),Y

Y-NT(n=30),andYY-TT(n=23).

5NY-NT,NY-TT,YY-NT,andYY-TT

(n=6).



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ets in the YY-NT group ha a greater ratio than ipigets in the NY-TT or YY-TT group, an on 14,pigets in the YY-NT group ha a greater ratio thanpigets in the NY-NT or YY-TT group.

Leukocyte Function and RNA Expressionof TLR

Percentage of ces positive for fuorescence resut-ing from bea phagocytosis i not iffer (P> 0.10)among the treatments or by ay (ata not shown).Fuorescence from CD14 expression was affecte byay (P= 0.04) but not by treatments. Fewer ces ex-presse CD14 on 1 an 7 than on 4 an 14. Thepercentage of ces positive for CD18 fuorescence wasaffecte by ay (P= 0.001) but not treatment. Moreces expresse CD18 on 14 than on 1, 4, or 7 aftertransport.

In jejuna tissues, a ay effect was etecte (P =0.07) for TLR4 (Figure 1). No main effects or interac-

tions were observe for expression of TLR2 or TLR4in iea tissues (P> 0.10). Day effects were etectefor RNA expression of TLR2 (P= 0.01) an TLR4 (P= 0.01) in MLN. Yeast transport interactions wereetecte in MLN for TLR2 (P= 0.001) an TLR4 (P= 0.08) expression. Aitionay, a yeast transport ay interaction (P= 0.09) for TLR2 in the MLN wasobserve. For both TLR2 an TLR4 expression, theYY-NT treatment ha the greatest peak expression on 1 after weaning, but NY-TT an YY-TT treatmentsaso ha greater expression of TLR2 compare with theNY-NT treatment.

Microbial Analysis

Lactobacii, tota coiforms, an E. coliwere enumer-ate from jejuna, iea, an ceca contents (Tabe 5).Lactobacius counts were affecte by ay in the jeju-num, ieum, an cecum such that counts were greater (P< 0.001) on 4 an 7 compare with 1 an 14. Tota

Table 4. Effects of yeast cuture an transport by ay on hematoogy1 for pigs fe a yeast cuture or the carrierony from 3 to 35 an transporte for 6 h or not transporte at weaning ( 21) in a 2 2 factoria arrangementof treatments2

Item HCT, %WBC,3

106/mLNeutrophis, 106/mL

Lymphocytes, 106/mL

Monocytes, 106/mL

Neutrophi:ymphocyteratio

Day an treatment 0NY 33.6 2.4 10.1 2.4 6.4 1.8 2.3 1.8 0.34 0.4 4.42 3.4

YY 32.2 2.4 11.2 2.4 6.0 1.8 4.0 4.3 0.25 0.2 3.53 3.2 1NY-NT 25.8 1.85 11.1 0.80a 6.0 0.46 4.4 0.37 0.36 0.04 1.46 0.10ab

NY-TT 28.4 1.85 10.4 1.80ab 4.8 0.46 5.0 0.37 0.25 0.04 1.04 0.10b

YY-NT 30.3 1.85 9.3 0.80ab 5.4 0.46 3.4 0.37 0.20 0.04 1.81 0.10a

YY-TT 26.1 1.85 7.7 0.80b 3.9 0.46 3.4 0.37 0.27 0.04 1.16 0.10b

4NY-NT 27.9 1.77 11.4 0.98b 4.9 0.56 4.9 0.45ab 0.24 0.05 1.03 0.11NY-TT 33.2 1.77 13.5 0.98a 5.7 0.56 5.9 0.45a 0.39 0.05 1.00 0.11YY-NT 14.5 1.77 16.9 0.98a 7.1 0.56 7.4 0.45a 0.38 0.05 1.19 0.11YY-TT 26.1 1.77 7.7 0.98b 3.9 0.56 3.4 0.45b 0.27 0.05 1.16 0.11

7NY-NT 29.5 2.26 12.2 0.98 6.4 0.56 5.1 0.45 0.41 0.05 1.39 0.12NY-TT 31.2 2.26 14.2 0.98 8.0 0.56 5.0 0.45 0.44 0.05 1.77 0.12YY-NT 28.2 2.26 10.9 0.98 5.0 0.56 5.5 0.45 0.28 0.05 1.08 0.12

YY-TT 30.2 2.26 15.0 0.98 7.0 0.56 7.0 0.45 0.58 0.05 1.11 0.12 14NY-NT 30.4 2.40 15.3 1.04 6.5 0.60 7.4 0.48a 0.69 0.05 0.86 0.13b

NY-TT 35.6 2.40 18.4 1.04 9.1 0.60 6.7 0.48ab 1.16 0.05 1.54 1.13ab

YY-NT 31.3 2.40 15.4 1.04 8.9 0.60 4.2 0.48b 1.05 0.05 2.23 1.13a

YY-TT 35.5 2.40 12.7 1.04 4.9 0.60 5.3 0.48ab 1.20 0.05 0.90 0.13b

Effect, P-vaueYeast 0.04 0.17 0.08 0.15 0.23 0.87Transport 0.40 0.66 0.30 0.81 0.01 0.06Day 0.001 0.01 0.01 0.01 0.01 0.04Yeast transport 0.36 0.01 0.02 0.49 0.84 0.02Yeast transport ay 0.87 0.04 0.10 0.02 0.18 0.04

a,bMeans with ifferent superscripts within a ay for each variabe iffer (P< 0.05).1Nontransforme means SE.2HCT = hematocrit; WBC = white boo ces; NY = contro iet; YY = yeast cuture iet (XPC, Diamon V Mis Inc., Cear Rapis, IA);

NY-NT = contro iet an not transporte; NY-TT = contro iet an transporte; YY-NT = yeast cuture iet an not transporte; YY-TT= yeast cuture iet an transporte.

3Vaues were square root transforme for statistica anaysis.

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coiform counts were affecte by ay in the jejunum (P= 0.04) an ieum (P= 0.03) such that 1 ha smaercounts than 7. Tota coiform counts in the cecum (P= 0.059) ha a ay effect such that counts were ess on

1 than on 7. Escherichia colicounts in the jejunumtene (P = 0.06) to iffer by ay such that countswere greater on 1 compare with 14. On 1, E. colicounts in the ieum were greater (P= 0.007) compare

Figure 1. Reative abunance of To-ike receptor (TLR) 2 (panes a, c, an e) an TLR4 (panes b, , an f ) compare with the 18S internastanar from tota RNA extracte from jejuna tissues (panes a an b), iea tissues (panes c an ), an mesenteric ymph noes (panes ean f) at weaning ( 0) an 1, 4, 7, an 14 after transport. Treatments were a factoria arrangement of iet an transport. Pigs were fe yeastcuture (YY; XPC, Diamon V Mis Inc., Cear Rapis, IA) or contro (NY) iets from 1 through 35, an one-haf of the pigs on each of thoseietary treatments were transporte for 6 h (YY-TT an NY-TT) or not transporte (YY-NT an NY-NT) at weaning ( 21). Data for ieaTLR2 an TLR4 an for jejuna TLR2 were og-transforme for statistica anaysis. Yst = yeast; Trn = transport.



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with those on 7 an 14. Escherichia colicounts in thececum were greatest (P= 0.002) on 1.

Isoation of Salmonellafrom MLN an ceca contents(P> 0.05) i not iffer among treatments. However,across a treatments, ceca Salmonellaiffere (P 0.10) the number of bans in the jejuna, iea, orceca contents after transportation (ata not shown).

A ay effect (P= 0.066) in number of bans in cecacontents was etecte such that ban numbers on 1were greater than those on 14 (22.9, 19.0, 19.5, an17.6 bans for 1, 4, 7, an 14, respectivey). Numbersof bans over a posttransport ays were simiar forjejuna an iea contents an were ess (P = 0.001)than in ceca contents (11.6, 11.1, an 19.8 bans, re-spectivey).

Simiarity coefficients of the DGGE ata were morecompex, with significant ifferences (P < 0.05) be-tween ietary treatments, transportation, an ays af-ter transportation, as we as interactions (Tabe 7).Simiarity coefficients on 0 were east (P< 0.05) inthe jejunum an increase from 55.1 to 71.9 an 70.7in the ieum an cecum, respectivey. Simiarity coeffi-cients were numericay greater than in our earier work(Wiiams et a., 2008), but in both stuies, simiar-ity coefficients increase aong the igestive tract (P< 0.05). Means of simiarity coefficients for ay, yeastsuppementation, an transport vaues within the jeju-num an ieum were not ifferent (P> 0.10) an were

ess (P< 0.05) than vaues for the cecum (52.8, 51.6,an 67.8, respectivey). Day effects (P < 0.05) weresuch that simiarity coefficients ecrease between 0 an 1 irrespective of iet or transport status, anthen increase over time towar the initia 0 vaues,athough there was consierabe variation in response(Tabe 7). Over a ays, simiarity coefficients for thecontro treatments (NY-NT an NY-TT) were greater(P= 0.029) in the jejunum than for the pigs fe yeast(YY-NT an YY-TT), whereas simiarity coefficientswere greater in the ieum (P< 0.001) an cecum (P=0.058) for pigs fe yeast.

DISCUSSION

Pigets experience severa stressors that accompanyweaning: a new environment, mixing with unfamiiar

Table 6. Effects of yeast cuture an transport by ay on the pig mesenteric ymph noe (MLN) an ceca Salmo-nellaoccurrences1 (n = 6 pigs/treatment per ay)

Tissuean treatment2

0 1 4 7 14

Positive Negative Positive Negative Positive Negative Positive Negative Positive Negative

Cecum

NY-NT 3 3 3 3 1a

5 6 0 5 1NY-TT 3 3 2a 4 6 0 6 0YY-NT 3 3 3 3 1a 5 6 0 6 0YY-TT 2 3 4b 2 6 0 6 0

MLN3 NY-NT 3 3 6 0 5 1 5 1 6 0NY-TT 5 1 5 1 4 2 5 1YY-NT 2 4 5 1 6 0 6 0 6 0YY-TT 2 3 6 0 5 1 5 1

a,bMeans of ceca tissue positive for Salmonellawith iffering superscripts within a ay iffer (P= 0.07).1Positive = positive for etection of Salmonella; negative = negative for etection of Salmonella.2NY-NT = contro iet an not transporte; NY-TT = contro iet an transporte; YY-NT = yeast cuture iet (XPC, Diamon V Mis Inc.,

Cear Rapis, IA) an not transporte; YY-TT = yeast cuture iet an transporte.3Within the MLN, a ay effect was etecte for a treatments (P< 0.05).



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pigs, materna separation, a new source of nourishment,

an sometimes transportation. Exposing the piget to anutrient either through the sow iet (K. Scott, PurueUniversity, West Lafayette, IN, unpubishe ata) orthrough irect suppementation (Bruininx et a., 2004;Mei an Xu, 2005; Kuer et a., 2007) is a methothat has shown some promise in aeviating part ofthe weaning stress. Using a S. cerevisiae fermentationprouct to ai in the ietary transition of weaning anstressors that often accompany weaning was the focusof this stuy.

We observe ony imite effects of yeast on BW be-fore weaning, which is consistent with the growth re-sponse to yeast in other stuies with pigs for the first 2

wk after weaning (van er Peet-Schwering et a., 2007)an with airy caves (Magahes et a., 2008). Trans-port effects were the ony effect on BW after weaningin this stuy. Aitionay, ADFI was affecte ony bytransport. However, neither van er Peet-Schwering eta. (2007) nor Magahes et a. (2008) provie yeastcuture uring sucking, as was one in the presentstuy. Athough pigs fe yeast ha greater BW gain inthe first 3 wk, those ifferences were abrogate by theen of the first week postweaning. By the en of thestuy, fee intake was more simiar, impying ony brief

effects of transport an iffering effects of yeast in ry

fee than in iqui fee. We foun that ony transportecrease ADFI in the first week after transport anweaning, but it i not change G:F. However, Carsonet a. (2005) foun greater ADG an ADFI when nurs-ery pigs were fe yeast extract, but this i not trans-ate into improve G:F. Simiary, Shen et a. (2009)foun improve ADG an ADFI when pigs were feyeast cuture. Part of that improvement was attributeto changes in gut morphoogy an immune mouationsrefecte by pasma IFN- concentrations that wereess than in contros at 7 postweaning but that weregreater than in contros at 21 after weaning.

In this stuy, changes in immune ce popuations

over time were reate to yeast cuture treatment antransport. Transport, together with yeast suppemen-tation, suppresse neutrophi an WBC counts thatwou typicay increase after a stressor. This stuyha weaning stress foowe immeiatey by transportstress, which create a compoun stress situation typi-cay seen in prouction. Transport treatments aso e-crease the neutrophi-to-ymphocyte ratio eary, anwith yeast treatment it remaine at a smaer ratio at 14. Age-reate increases in CD14 an CD18 expres-sion were evient, but no treatment ifferences were

Table 7. Denaturing graient ge eectrophoresis simiarity coefficients of jejuna, iea, an ceca bacteria speciesin pigets1

Tissue an treatment 0 1 4 7 14 Cross-prouct2 SEM

JejunumNY-NT 56.23 40.64c,z 62.75a,x 56.80a,y 62.00x 50.72y 4.17NY-TT 60.61a,xy 43.94c,z 66.05a,x 56.01xy 53.65y 10.2YY-NT 53.93 35.65c,z 48.21bc,y 56.96a,x 55.44xy 48.22y 5.89YY-TT 44.74bc 56.24ab 46.46b 51.11 48.64 5.89

Cross-prouct 61.63 48.72b

50.09bc

56.51a

56.00SEM 6.25 5.89 4.17 5.89 10.2

IeumNY-NT 65.83b 44.81b,yz 38.53c,z 37.67c,z 59.94x 44.90y 4.26NY-TT 54.66a,x 55.19ab,x 43.87bc,y 49.21xy 49.39xy 6.02YY-NT 77.92a 57.11a 54.30b 56.50a 56.82 55.07 4.26YY-TT 47.85ab,y 64.18a,x 46.32b,y 58.74x 46.51y 3.30Cross-prouct 64.46b 52.92ab 52.35b 46.56b 58.02SEM 2.61 4.26 3.30 3.30 6.02

CecumNY-NT 69.89 62.20bc 68.32b 67.06c 66.59b 65.32 3.52NY-TT 57.86c,y 78.52a,x 69.03bc,x 60.00b,y 61.39y 6.10YY-NT 71.96 70.55a,xy 66.67b,xy 77.76ab,x 74.93a,x 66.55y 6.10YY-TT 66.67ab,y 63.44b,y 83.08a,x 66.07b,y 65.86y 6.10Cross-prouct 75.47 65.26b 68.17b 73.78b,c 67.47b SEM 3.07 6.10 6.10 3.52 6.10

acMeans within coumns of a tissue with ifferent superscript etters are significanty ifferent (P< 0.05).xzMeans within rows of a tissue with ifferent superscript etters are significanty ifferent (P< 0.05). The NY-TT -4 vaue of 43.94 was

significanty ifferent from a the other vaues, an thus the z esignation. The xy esignation for the NY-TT -14 vaue of 56.01 inicates thatit is simiar to the -1 vaue of 60.61 an is aso simiar to an between the -7 vaue of 66.05 an the cross-prouct vaue of 53.65, which arestatisticay ifferent. In the cecum for 4, the NY-TT vaue of 78.52 is significanty ifferent from a other -4 vaues, except for the YY-NTvaue of 66.67 (because of greater variation within these vaues), even though the YY-NT vaue is ess than the NY-NT vaue of 68.32.

1Least squares means. NY-NT = contro iet an not transporte; NY-TT = contro iet an transporte; YY-NT = yeast cuture iet (XPC,Diamon V Mis Inc., Cear Rapis, IA) an not transporte; YY-TT = yeast cuture iet an transporte.

2Cross-proucts are comparisons of iniviua sampes within an between treatments an ay.

,Because of arge SE within one of the treatment sets, means within rows or within coumns of a tissue, respectivey, were not ifferent (P>0.05).

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etecte. Custer of ifferentiation 14 is part of theipopoysaccharie receptor that functions in interac-tions with an ientification of gram-negative bacte-ria, such as E. coli an Salmonella. The istributionof monocytes, macrophages, an enritic ces aongwith the frequencies of CD14 expression on those cesin intestina an systemic ymphoi tissues respon tointestina coonization by actic aci bacteria ifferenty

than oes an enteric pathogen such as the human ro-tavirus (Zhang et a., 2008). Lactobacii counts as weas the CD14 expression on immune ces were not if-ferent among treatments. Lipopoysaccharie biningcan be accompishe through other substances founin porcine mik (Shahriar et a., 2006), such as acto-ferrin, soube CD14, an serum amyoi A. However,ony actoferrin cou withstan igestion (Lee et a.,1998). Serum amyoi A concentrations were foun toincrease significanty in oer pigs after a ong trans-port (Pieiro et a., 2007). Therefore, in this stuy, ifipopoysaccharie was being controe by moecuesother than ce surface-expresse CD14, expression of

ce surface CD14 may not have been stimuate.The ack of phagocytic ce response is curious. In

aition to neutrophis, a subpopuation of eukocytes,natura kier (NK) ces, are the preominant innateimmune ces for pigs (Pintari an Saamer, 2008).These ces an T ces typicay express the CD8 cesurface protein (Denyer et a., 2006). Feeing yeast cu-ture to nursery pigs ecrease the CD4:CD8 ratio (Shenet a., 2009), impying that yeast cuture enhances theNK ce or T ce popuations. In vitro work (Jen-sen et a., 2008) showe that yeast cuture enhanceexpression of CD69 an CD25. Increase ymphocytenumbers, in aition to neutrophi popuations, may

be refective of NK an cytotoxic ce activity in thisstuy. That occurrence may have resute in ess neu-trophi stimuation an reuce CD18 ce surface ex-pression an phagocytosis by the neutrophis.

Intestina microbia ecoogy iffere for E. coliin thejejunum in the current stuy. Previousy, E. coliconcen-trations in the cecum, but not in the coon or rectum,were reuce by feeing a yeast cuture to weaning pigs(Shen et a., 2009). When aggutination of pathogenicbacteria by a yeast extract was teste, 64% of E. colistrains that were teste an 67% of Salmonella spp.strains that were teste were foun to aggutinate tothe yeast prouct (Kogan an Kocher, 2007). Salmo-nella entericaserovar Typhimurium strains (70%) anS. entericaserovar Enteritiis (86%) were even greater.The authors cite 3 mechanisms for the effect of theyeast prouct on pig heath: 1) inhibition of pathogenahesion to gastrointestina epitheia tissue, 2) stimu-ation of immune ces in Peyers patches, or 3) asorp-tion of mycotoxins an inhibiting their actions, or a 3.

Diet ha no effect on the number of preominantbacteria (bans) in the various sections of the intestinatract. This is not surprising in that the yeast prouctwas not fe at the substrate eve. Even though there

was no iet effect on the preominant bans, the atai not rue out an effect on ess ominant bacteriapopuations that affect pig heath. Davis et a. (2007)saw an increase in the number of bans 20 after wean-ing in pigs fe an antibiotic compare with contro pigs,but no time course was aresse. There was aso noeffect of transportation, which is somewhat surprisingconsiering that stress is thought to isturb the gas-

trointestina tract an aow for fuctuation in bacteriapopuations in the intestina tract. However, one mayhave fuctuation in bacteria species an sti have thesame number of preominant bacteria. There was a ayeffect in ceca contents, in which the number of banswas greatest at 1 an ecine thereafter. Konstan-tinov et a. (2003) foun a ifference in the numberof bans after weaning, as in the present stuy. Theincrease in the number of preominant bans in cecacontents is consistent with the we-known increase inbacteria numbers an species in the cecum comparewith the jejunum an ieum.

Even though the number of preominant species

increase from the jejunum to the cecum, suggestingmore compex microbiota, the simiarity coefficientswere greater in the cecum than in the jejunum, sug-gesting that the igesta fux in the proxima sma in-testine maintains a greater iversity of bacteria in thejejunum compare with the cecum. Numericay, the 0 simiarity coefficients were greater in a sections ofthe intestina tract than at any ay postweaning. Post-weaning, simiarity coefficients were east at 1 angeneray increase over time, suggesting that weaningmay cause a isruption of the intestina microbiota.The greater simiarity coefficients in the ieum an ce-cum, but not the jejunum, for pigs fe yeast may sug-

gest that the yeast has ess effect in the jejunum. Eitherthe increase fux rate or the greater microbia iversi-ty (smaer simiarity coefficient) in the jejunum causesthe reuce response to the yeast prouct. This is incontrast to the ata of van er Peet-Schwering et a.(2007) in which they observe a ecrease in iversity inpigs fe yeast. The ata emonstrate that weaning oescause fuctuation in the preominant bacteria popua-tions in the intestina tract. An interesting question iswhether there is a greater effect on nonpreominantbacteria popuations, such as coiforms, that may havesignificant effects on anima heath. However, our atashowe ony ay effects on tota coiforms.

Overa, many compex interactions were etecte.However, a time effect persiste across most variabes.We first saw increase TLR2 an TLR4 in MLN on 1. That response was greatest in pigs in the YY-NTgroup, suggesting that the yeast cuture ha primeimmune ces for the changes that occurre with wean-ing. By 4, Salmonellaspp. ha increase in the MLN,an by 7, cecum Salmonellaspp. ha increase. Theimmune response of increase monocytes was observeby 7 an 14. There were many transport an yeastinteractions with ay. However, the most striking ef-



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fect of the combination was reuce WBC, specificayreuce neutrophis.

The yeast treatment ha a prouction response sim-iar to yeast cuture treatments in other stuies. A-though much variabiity in responses epens on size,-gucan origin, an extraction processes (Sonck et a.,2010), it appears that proviing a smaer fragment ofa specific portion of the yeast, as in a yeast extract,

provies a ifferent response than when the whoe yeastcuture is fe (Kogan an Kocher, 2007). Aitionay,the quaity of the housing environment an the stressorsthat were experience cou expain some of the iffer-ences that were note between this an other stuies.Pigets in our stuy i not have to compete for feeor ominance after weaning because of the iniviuahousing. Athough they ha visua, auitory, an sometactie access to other pigets, the reative isoation mayhave been a negative factor. Overa, the resuts suggestthat yeast cuture has immunomouating propertiesthat may ater interactions with intestina microbiaecoogy. However, more compex stuies are require to

euciate this reationship. It appears that eary eiv-ery of a yeast suppement to pigets has some benefits.

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