Endogenous ileal amino acid excretion in monogastric animals

166 Pages·2012·4.1 MB·English

by  Butts, Christine Ann

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Endoge usI leaAlm inoA cid
�xcretiionnM onogastrAinci mals
A thespiress entienda rtpialf ulfi!m.ettt
oft l1ree quitemfeontrti sdl eeg roefe
D0:cto,r ofP hilosoipnhA yn imaS�c ience
atM asseUyn ivers-ity
ii
ABSTRACT
A new metfhocxlrt hdee terminaotfie onnod genoiulse aamli no aecxicdr etion
undecro nditioofpn esp tidael imentawtaisro enfi neadn de vlauatewdi tsht udies
involvtihnelg a borartaotryT. h ere fmedm ethcxlw asu sed itnov esttiega aspecotfs
endogenioluesa l aamciinfdol owi nt hep igF.i vset udiweesr e condutchtreede,
witrha ts tawnoid n volvtihngrego winpgi g.
1.P reliimnaryi nveisgttaionesva luatetdh ef ilrtatieofnif cienocfy
ultrafiltradteivoinc aensd,e xamintehdr eper e-filtrattrieoanet nmtfso rr ait leal
digesttrai:c hloroaacceit(diT cC A)a ndp ercohrliacc i(dP CAp)r eciitpatiaonnd,
centrifugatio(nS PIN)T.h er ecoveorfny i trogfeolnl owiunlgtr afilatrtio(nm olecular
weigehxtc lusliiomn1i t0, 000D altonsfi)f toefpe unr ifpireodt epienp,ti daen d amino
acisdo lutioinnsd icaatnee ffde ctivfiel tration (>o9n0n %o)m inmaoll ecruw leaight
byt heu ltraifltratdieovnie cs.D eterminaotfin oiontrg ena nda minaoc idisnt he
resultfriancgti onfsol lowiTnCgA andP CA precipitaatnidco enn trifugatioon fr at
iledailg esitnad icattheaPdtC A wast hem osetf fectipvreec ipitEanndto.g enous
ileal amino aciidn t ehxegcr rowenitigro ants afnee dn zymichayldloryl ysceads ein
(EHC)b ased wdiiteshtu bsequetrneta tmenotft hed igesta tuhseui lntrga filtration
tecohlnogwye ret hedne terminTewde.l v1e00 g male rwaetresf ede ithaenrE HC­
baseddi eotr par oteriene-d fieatn ds ampleosf d igeswtear ec olleedca tfter
sluaghetr.T hed iegsat fromt he6 EHC-fedr atwse reu ltraiflteedra fter
cetrniufgatioann dt hhei gmho lecuwleairg hfrtac tiaodnd etdo t hep reicpitaTthee.
protne-rifeef edr athsa ds ignainfitcl(yP <0.0l5o)w earm inaoc ifdl owtsh atnh ose
ratfsed thEeH C-basdeidew ti tshu besquetrneta tmenotf t hdei gesta.
2.T hep rooprtioonfes n dogenporuost eipne-p,t idaen-d fraemei naoc id
nirtoge(nN )i nd igsetNa fromt hed itsaill euomf t her ati mmeidatelayf ter
collieocnot rf ollowisntgo rafrgoez e(n- 20a°nCd - 916°Cw)e rec ompareEdi.g hteen
growirnagt wse reg ivean p roetinre-ef diefto r6 days,e uthanaasnedsd am pleosf
digeswteare c ollecfrtoemdt hete rmin2a0l comf i leumTh.e s toraogfde i gesdtiad
nots ingificanatfelfyct th epr oprotionosfN -contaisnuibtnsagn cienst hep recipitate
plurest entaotreu ltrafiltrfartatieco nsO.n avera,g6 e7%o ft hteo tal dNig weasst a
int hep recipiptlautrsee tentfaratcet iaonnd 3 3% otfo tdail gesNt waa s tihne
ultrafiltrafrtaec tionF.r eaem inaoc iNd a ndp eptidNe w ere1 0.4 a1n0.d6 %o ft otal
digesta N, respectively.
3.T hee fefctof u sindgi fefrenfltu sihngm edifao rt hec ollecotifoi nl eal
digesotna tchoem posoinoti fe ndoegnouNs w ase xamineTdw.e lvgreo wingr ats
wereg ivean p roeti-nfredei eatn ds amlpes iolfe dailg eswtear e collfreocmt tehde
iii
euthanaansiemudas lie nigt dhiesrt illoerpd h ywsaitoelsroa gliaicsnta ehfll eu shing
mediuTmh.e rwea nso s ignifiecfafnoetfcc otl lemcetitohnod t holene vels
ofN -co�ntgs ubstiannr caeetns d ogeinloeduaislg esta.
4.T hee ffeocfst tsao tfbe od y nitrboagleanan nctdeh per eseonfdc iee tary
peptiadnedps r otieni ndt ihgee strtaicovtnet heex creotifeo nnd ogeanmoiunso
acids thfrieol me ouftm h pei wge rien vestigEantdeodg.e nous lysine excretion was
determipniegdgs i vfaeop nrr oteidni-eafrtnEe, He C -az, e ionra- s ynthaemtiinco
aicdb-asdeide Etn.d ongoeufsl ofwosar m inaoci dost htehra lnyi snwee re
determfionre dop nit ghpser ot-efrieanen EdH C-badsieedSt sim.xa le (p1ik5gg s
livewewiegrahelt l)oc attoee adco hft hfeo ur direectesi tvahednedid fe ot1r 0d ays.
Them eaenn dogeinloeluaysls ifnleof wostr h zee iann EdH Cf epdi gwse rneo t
significdainftflebyru weten reth igh(ePr< 0.t0h5at)nh oset hpferoo tr eina-nfrede
synthaemtiinacoc id fewdh opsmieeg asfln o wwse re sniogtn ificdainft1_1fltey r e
froema coht hTehre.m eaenn dogeinloefluaoslw f so arm iancoi odtsh tehral ny sine
were highefro tr(h EPeH< C0f .e0pd5i )gc so mpatreotd h e anitmhapelr so toeni n­
free edxiceeftpop,trr ol,gi lnyecainnader ginine.
5.T hee ffoefcf to oddr mya ttienrt aoknee n dogeinloeuaasml i naoc id
excreotfti hopeni ugn der paelpitimdeenw taasdti eotne rmiSniexdt.me aelnpe i gs
(50 lkigv eweeiagcfihhtt t)e da Tw-ictahn niuntl hae teirlmeiwunemar lfee adt8
levoeflf sood drmya ttienrtf aokpree rioodf8s d aysT.h ee xperiimnevnotlt vweod
triaolf8s p igesa ccho,m priasc irnoogsv sed re siEganc.trh i ailn vol4 vpeadi rs of
pigwsi teha ch rpeacieri ovnieon f4g s equeonfctre esa tmeEnatc.she quence
compsre4id l evoeflf so oddry mattienrt aakrer anignae L da tsiqnu aTrheef. o od
drmya ttienrt awkeerse 0.06, an0d0. .018a2,n, 0d 0. .0015.0,0 07.,0a 9n 0d. 11
0.75 -1
metabloilviecw eciwgh)td ayfotrh fier satn sde cotrnida relssp,e ctively.
Therwea as ni ncrienia lseeea xlc retioofan m iancoi ndistr,o agneddnr mya tter
with
increafsoiodndr gmy a ttTehre.r e swiegrnei fi(cPa<n0t.l 0i5n)re ealra tionships
betweeennd ogeinloeauamsli naoc iadn ndi trogeexnc reatnidfo onod dry matter
intaekxec efpoltry signleu,t aamciiacdn pdh enylawlhainciihnn cear seeidna
curvilmiannenaeTrrh e.s ree latiodnesthenirepmudsin ,d pehry siolomgoircea lly
normcaoln dittihoanunsn dperro teianl-ifrmetieeon ntp,ar ovpirdeel imdiantoarany
thmea gniotfus dmeai lnlt esatimnianaloc ildo sisnet shp ei g.
lV
ACKNOWLEDGEMENTS
Is incertehlanyk m y supervisoDrrsP ,a uMlo ughaann dA ssoactie
ProfesBsiolrSl m ithf,o rth eirg uidanacned e ncouragemtehrnotu ghout
thisst udy.
I am alsgor ateffuolr twhiel lisnugp poarntd e xpertoifs eD r
Gordon ReynDorlD dasv,i Cda rarn dD rC am Reidf ort hes urgearnyd
pos-toperatciavreoe f t hep igsT.h ankasr ea lsdou et o:M s Felicity
JacksoMni,s sR osemarWya tsonM,r sG eraldiWnoeo d,M r JuliRaeni d,
andM rsC aroFll ygefro rt heiars sistawnictehc hemicaanlay lsesM;r s
IrenHea ll afsors istadnurcien sgu rgerMyr; BarrPya rlaanned Mr
GrahamP earosnf ort echniacsasli stanDcreD ;o riaGna rriacnkd M r
CarlosS osaf ora dviocens tatisatniacylasle s.
Iw isht oa cknowleThdeg eN ew ZealanPdo rkIn dustrByo arfdo r
fundintgh er eesarch.
I wast heg ratefrueilcp ieonfta MasseUyn iversiGtrya duate
Assistantasnhdia p M asseUyn iversViitcyCe h ancellSotrusd Gyr ant.
The guidanocfet heM asseUyn iversAintiym aElt hicCsomm ittee
ina ppronvgit he experipmreonttoaclio nlvso lvtihneg uosfae n imailss
acknowledged.
Thec ompanionsahnidap d vicoefA lisoDna rrgaha ndo thepro st­
graduteas tudenatnsd, stmaefmfb e·sr oft heD epartmeonftA nimal
Sceincei sg ratefualclkyn owledgFeidn.a llIwy i,s ht oe xpremsys
fondeasptp rieactitoon mhyu sbanMdu rrafyo rh isl ovinsgu pport,
encuoragemeanntda ssistatnhcreo ughtohuists t udy.
CONTENTS
PAGE
Abstract
n
Acknowledgements iv
LiosTfta bles
IX
Introduction 1
Chapt1e r
RevieowfL iterature 3
1.I1n troduction 3
1.P2r otDeiignes tainodAn b sorption 3
1.2G.a1s triDci gestion 4
1.2D.i2g esitinto hnSe m aIlnlt estine 4
1.2.3 MDuicgoessatli on 5
1.2A.b4s orpotifto hPner oducotfPs r otDeiigne stion 5
1.2D.i5g esint tihoLena rIgnet estine 7
1.G3 astrointesPtriontSaeelic nr etions 8
1.3.1 SaliSveacrrye tPiroont ein 9
1.3G.a2s triPcr otSeeicnr etion 10
1.3.P2h.y1s iolCoognitrocofGal al s trSiecc retion 11
1.3.E2f.f2oe fcA tg ea nDdi eotnG astrPirco tein
Secretion 12
1.3P.a3n crPeraottiSecei cnr etion 13
1.3.P3h.y1s iolCoognitrocofPal al n crSeeactriect io1n4
1.3.E3f.f2oe fcA tg ea nDdi eotnP ancrPeraottieci n
Secretion 15
1.3.D3i.e2t.parary o tein 15
13..3.D2i.ebtf aaartny sd t arch 16
·
1.3.D3i.e2t.fiacbr rye 17
1.3.R3a.t2oe.fa d d aptoaftip oann crseeactriecti on
tod iet 18
1.3.S3u.m2m.aeor fpy a ncraedaatpiitoctan odt iet 18
1.3B.i4lP er otSeeicnr etion 19
1.3.P4h.y1s iolCoognitrocofBal il l See cretion 21
1.3.E4ff.e2oc ftD i eotnB ilSee cretion 21
vi
PAGE
�.3I.n5t esStiencarelti on 22
1.53..B1r uBsohr dEenrz ymes 23
1.3P.l6a sPmroat eSienc retion 24
1.3E.p7i thCeellLoilsa sl 24
1.3M.i8c robPiraolt ein 25
1.3M.u9c uSse cretion 26
1.T4o tQaula nttiioefsE ndoegnouNsi troagnedAn m ino
AcidSsec retIendtt oh e GastroTirnatcestt inal 29
1.D5 iegstioofEn n dogenPoruost Seeicn retions 32
1.D6 eterminaoftE indoong enNoiutsr ogaendnA minAoc id
ExcretiinDo ena lg esDti-aM ethodology 34
1.6P.r1o tein-Free Method 35
1.6.2 SAymnitnAhoce iDtdii ect 38
1.36R .egressMieotnh od 38
1.6H.o4m oargMientihnode 39
1.T6r.a5c Meert hod 42
1.6P.e6p tiAdlei nmteation 43
1.C7 onclusion 44
1.R8e ferences 45
Chapt2e r
EndogenoAumsi noA ciFdl owa tt heT erminllaelu m
oft he Rat DeteUrnmdienCreo dn ditioonfPs e ptide
Alimentation 69
�lA�tract W
70
2.I2n troduction
72
2.E3x eprimental
2.3V.a1l idoaftU ilotrna fiTletrcahtinoonl ogy 72
2.3E.v2a luaotfPi roen- FiTlrtreaatitomne nts 72
2.3.C2o.l1l eocfIti loenDa ilg esta 72
2.3T.re2a.tm2e notf Digesta 73
2.3.F2r.a3c tioonfta hStieuo pne rnatants 73
2.3D.e3te rmionfEa tinodno geAnmoiunAsoc iFdl ows 74
2.3C.h4e miAcnaall ysis 74
2.3D.a5tA an alysis 75
vii
PAGE
2.R4es uls t 75
2.4V.a1l idaotifUo lntrafil tratiMoent hod 75
2.4P.r2e -filTtrreaatitmoenn ts 75
2.4E.n3d ogeAnmoiunAsoc iFdl ows 77
2.D5i scussion 79
2.5.1 VaolfUi ldtrafilatrtiaotinTo enc hnology 79
2.5P.r2e -FiTlrtreaatitmoeonnf tD igesta 80
2.5E.n3d ogeAnmoiunAsoc iFdl ow 81
2.R6e efrences 83
Chapte3r
ProteiPne-p,t iadned-F reAem inoA cid-Nitrino gen
EndogenoDuisg esNtiat rogaettn h e Terminal lleum
oft heR at 85
3.A1b stract 86
3.I2n troduction 86
3.E3x perimental 88
3.R4es ults 91
3.D5i scussion 95
3.R6e efrences 99
Chapt4e r
CompositoifoE nn dogenoDuesaD li gesNtiat rogFeonr
theR a-tTheU seo fD istiWlalteedfr o rD igesCtoal lecti1o0n2
u��rnct 1m
4.I2n troduction 103
4.E3x perimental 103
4.R4es ults 104
4.D5i scussion 105
4.R6e efrences 107
Chapte5r
EndogenoLuyss ianned A minoA ciFdl owast t heT erminal
lleumo ft heG rowinPgi g( 15k-g2l 5iv eweiDgehtte)r mined
Vlll
PAGE
UndePrr otein-SFyrneteh,e Atmiicn oA cidP,e ptiadned
ProteAilni mentation 108
5.A1b stract 109
5.I2n trotdiuocn 101
5.E3x eprimental 111
5.13A .nimals 111
5.3D.i2e ts 121
5.3E.x3p erimPreocnetdaulr e 121
53..C4h emiAcnaall ysis 141
5.3D.a5tA an alysis 151
5.R4es ults 161
5.D5i scussion 191
5.R6e efrences 172
Chapte6r
TheE ffecotf F oodD ryM atter InontE ankdeo genous
lleaAlm inoA ciEdx cretDieotne rmiUnnedde Cro nditions
ofP eptiAdlei mentaitn�t ohne5 0k gL iveweiPgihgt
6.A1b stract 123
6.I2n troduction 123
6.E3x perimental 143
6.13 A.nimaanlHdso using 143
6.3S.u2r gIimcpalla notfSa itmipoln e
T-canniuntl hTaees r miinlaelu m 143
6.3D.i3e atnsFd e eding 143
6.3E.x4p erimPreocnetdaulr e 153
6.53 C.hemiAcnalal ysis 163
63..D6a tAan alysis 173
6.R4e sults 173
6.D5i scussion 183
6.R6e ferences 124
Summarayn dC onclusions 174
IX
LISTO F TABLES
TABLE PAGE
1
1.1 Amounotfns i troge(ngd ay)-s ecreitnettdoh e
gastrointtreatsco tiftn ha5el0 k g livepwiegi ght 30
1
12. Endogenaomuisna oc isde cre(tigod nasy) -i nto
thgea strointtreascotitftn hapeli g 31
13. Endogenaomuisna oc isde cret(igdo an-y1s)r emaining
unidgesatneuddn absoarttb heed oeftn hdie l eumt hoef
pig 33
14. Summaryo f litvearlautfeuoserrn e d goenoiulsea amli no
-1
acid ex(cgkr getidornmy a ttienkrte ai)n t hpei g ·
determuinndepedrr o itner-efalei mentation 36
15. Summaryo ft hlei atteurvrael ufeoser n dogeinloeuasl
-1
aminaoc iedx cre(tigko gn dry materti ntaiknte h)pe i g
determbiytn herede egsrsimoent hod 40
-1
21. Inrgeidencto mposi(tgki gona idryr weigohftt )h e
enyzmicahlyldoyrl ysceads ein a(nEpdHr Con)t- eefrie
diets 73
2.2 Ther ecorvyoe fn irtoge(nN )for mp uirifepdr otein,
peptaindadem inaoc isdou ltioinnts hr eet entaantde
filtraafttueel rtr afiltration 76
2.3 Mean( ±SEe)n dogeanmoiunsao c ifdl oawtst htee rminal
ileoufmr at(sn =f6e)ad n e nzyamlilhcyyd lryosceads ein-
basdeided ett eirnmeodnt hteo tuanltr eated odri gesta
digesta stuocb ejneutrcigtfaetipdol nuu sl trafialtitron 78
2.4 Mean(± SEe)n dogeanmoiunasoc ifldo wast t htee rminal
ileoufmr aftesad n e nzymihcyadrlollyy csaesde( iEnH C)
baseddi edte termoinnd eidg essatmalp essu jbectteod
centrifugtiaonp luusl trafilatrnadt ifloonw s ffdeo ra rats
proteriedeni -eft 79
31. Ingrieedncotm pos(igtk ig-1o n drwye igohfta )
air
protfeeriedn ifeettd o ogwrinrga ts 89
-
3.2 Mean propoofrn tiitornno-sogc netaicnomipnogu nidns
thper ecipaintudal ttreaf iolfitr laetdaeil g efsotrraa ts