Fouda, T. A. ct al ••. ISSN 1110-7.~19

EFFECT OF SELENIUM DEFICIENCY ON OVINE IMMUNE STATUS, FREE RADICALS PRODUCTION AND ITS

CORRELATION TO DISEASE OCCURRENCE

Fouda, T. A.': El-Sayed. O. R.·· and Nasr. M. Y.· ..

1'13

Oc:panlnt'o{s of [nternal Medicine & InfcdJous Olsea('ies·, Biochemi,,!ry ,llld Blochemislry or Ilutrition··,

Faculty ofVelerlnruy Medicine. MansolJt'a Univer('iity and Depart! 'II or t\nllt'w.1 f,tC'dieinl· U.,

[<'acuIty ofVelerinary Medidne. Tantn Univcrsil /. 1'~i!.Yp[

ABSTRACT

Tilt.,.;; stud!} have been applied on a total oJA/tv sheep qI both sexes sdecled Jhllli

illrcl' dUJerent f\heep }locks, in order (0 declare OK> ~a(}C{s q{ sereJtfWll dc./kk'nqJ 011 (//('

immunc Ji..mdinn, production qr free radical .. and incidence of some disease ronlifliolls.

TI1C scfee/ed animal .. have oc"'(;!n allotted into tlVO groups, Group -1 involved those dint·

enUyand apparently healthy animal!> (n ""' !O) and tvere served as control group. While.

group -2 tnuolued those antmals (H :;:;: 401 that had (lie dinical sign oj emaclafioll. rough

coot and poor borfy condition,

Two blood sumptes (heparinized and nan·heparlniZed) have been oiJlnirK"!l .fmm

each animal in both groups and 1!Jelt:' sUQJccted to lht' Caborat(wy tm-estfgaliolls. 11),('

obtained biochemi.cm d.ai.a ret.>ealed a signij'<cant redHdion in the rn('nll values qr II Urif'.

oxide. T3 , total lipids and trig[yccrides associated Willl significant elevaliOl\:) in tilt'

'l1<--'an ~altjes oj Malortdialdchyde and T4 fn seieniwl1-dt;/kient sheep in colll[Jntison

wilh those oIhealthy ones.

Dclcrminatton oj the enzymatic activities particularly ca1ala. ... e, Stlperoxld(> dis 1Il1l­

lase and glucose-6·phosphnte dehydrogenase ltalle been carded out, and the oiJlnirlf'd

resulls revealed that selenium-deficient sheep had a reduced values oj tllese eIlX!}IlU;S

{f compared with those oj healthy ones. 'l11is reduction migllt be all:ributed to fhe otX',

consumption oj these antioxidants resuUedJrom increased levels qrtheJree radicals in

the cells of sdellium1ieftcierit sheep. Moreover, seleniwn·deJkiellt sheep had q lowe/'

oalucs Jor btood~sclenium, immunoglobtdLtts (JgM and 19GJ wilkh mati result in ill­

crL"a.. .... 1d risk when those dc:.r!Cient an/rna!s are exposed 10 nJurther disease problem.

In addition. ,dluiw- f'.xarnina(iou qf mitk samples rt."Vooted that, seieniulll d(:fidcfl(·

sheep had Q higher somatic cell counts tharl Ihost' oJ the heaUfllJ sheep, indicalillU

"Mansour«. Vet. Med. J, (173 . 186) -Vol. V. No. 1.-:?003

Fowl«. T. A. et «LH

some son qf a ~ltb-cUnical.lonn oj mas£itL ... in (hose sheep. Diet an.uysis re,->eak~ fila!

1Tl(' /He(In t!(!lues rd'selenium were t){!r)j low thun the reL-'OmmeIHlei t'Olues.lnr the Wli'

m(d~.

III nmdtlsion. selenium uejlciency is associated with decrea."i(;'u immwlOglolJUlins,

tllcr{.J(L"illlg Iree radicals productkm mith consequenl cellular damage and incn:."Ostng

risk qr diseases

INTRODUCTION

174

Sdcnilull is an css('nUal elcment for growth. rcproduciton. and rli"'l,.'USCS prcv('ntitHl ill all all\­

rnals. Its primary fnn~' lion Is to protcct reH mcmbrancs nnd proleiu8 from damaging ('licmicals

th,u arc formed during nonnat metubolJsm. Body selenium (,Ol1ccntraHons arc diredly r'CIaled to

dielary intake of sclen1mn in plant produt'ls normally fed 10 Uvcstock (Gates and Johnson.

2002).

Sekniulll is it component of the J!lutathlon peroxIdase enzyme, 1111s enzyme is imporlalli in

:lrn!edin~ tIl(' Intcgrlt¥ of cell membranes. During normal meiabo}fSITL the cell prodllces rC'a:::tlv('

(orlll:$ of oxygen ipt'Joxldes) which. Jf not aUcr('d. will dama~{' the unsaturated fatly <Kids JlJlllld

In the cell memhrane, Membrane damage will dIsrupt eell function and adversely ~,nC<:'! 1 h(, anl~

mal health (Gatee and Joltnaon.2002).

'This selenium-dependcnt enzyme catalyzes the reactions that aid in the rcdnclion or iJolh in­

,;rgaoic and organlc hydro-peroxides. Vitamin E and selenIum through the giuta!hiull l)t'rox\d.l5e

(;nzymc arc involved In Intracellular dcfensClj against oxjdant dnmagc (Debskl et .Ql •• 1992).

AI lcas( clc\'en selcIl.OpTOtclns have been charactcrt~d, and the."c i~ evidence that mJtHtlnllal

scienoproldns exist. M')st of the biologically active T3 (trHodothyronifw) in the cirClllafion and itlM

~ide ('ell::; is created by the removal of one iodIne atom from 1'4 (thyroxtn), In a reaction t'ataly/..cd

hy ~d('nJllm-depcndent iodothyronine dciodinasc enzyl1!cs three different scleniulIl-dcj>clltkni

iodo1hyrollinc dciodin:'!,.es (types 1, 2 and 3) can both activate and imu::tJvutc thyl'Old 11Ol'I1wnc,

making sdcnium an e~ntlal element lor normal developmcnt, gmwth, and met<)bo(j~m lhron~h

the IX'l;ulalion of thyroid hormones (Larsen.1998: and Holben &: Smith.1999).

Selellilull deficiency has been associated witlllmpalroo function of the Imrmmc system. Mon'­

qVCl", st'lclHum supplementation tn fndlvJduals who are not overtly selenium dctlclcnt appears 10

'lliull1lalc (11(' immUne response, It Is also indicated by many auUtors Lhal scleniutn plays a role

;n regulating the L"Xpression of ceil signaling molecules culled cytok1nes which on'hesll"ak lhe

Inunmw rcsl)(m~c. Selenium deficiency suppresses the Immune sy.'item responsc, inted('rlllg

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Foud~ T. A. et aL.

175

wH!I lis abllHy to fight. inleclion and lowering resistance to dl~K'ase, These problems Ii)!' (he new­

born (:811 mean the dIfference in lheir survival or death (Combs and Gray,199S).

Selenium affeets not only polymorph 11l1ch.'ar neutrophil func(kJll, but also lymphocy(\: pmlifc

mOon. cytn(oxlcHy, and anUbody produd[on (Grasso et at. 1990). Impaired bovine lYlllpilo<)'1c

function and signaling in response to selenium delkkncy luay ('ause ihe delaycd mUm, of poly­

morphnuclc<lr (PMN) cells to the mammary gland observed m severe clinical signs in st'lt'nimH

deficient animal challenged v-ith g, col! (Erskine et al,. 19(9). Pi, '1cr IgG cont'Clltralions also

were observed in the scleniulll·supplemen(cu ponIes Knight and T;;znlk (1990).

In addition, Selenium supplementaU(U) has been linked to low('r herd somatic eell ('OlllltS llnd

has been shown to decrease the prevalence and severily of dilliml mastitis IEcskine et

al .• 1989).

The f'ontcnt of selenIum in blood is a rdlable indicator of the selenium Slatn$ in nni!ll.th,; alld

jhe valm!s less than 0.04 ppm, 0.05 ppm, ami O.07ppm ltI bee!, sheep and ('ow's. rc~pe('livd.\'

were ['('ported as a dc1kient values (Combs and Gray, 199B). Ovinc uloud .sckllltHil l'Olwellti',\,

Lion less than 0.04 ppm are eonsidered diagnostic of selenium defiCiency (Whanger et aL, 1977),

Con~cqllcn!ly, this study was desIgned in order 10 dcclare the dTt't'is of selenllll1\ dd)('iC"twy

on the immune funcBon. production of free radicals ami occurrencc of somc disClUW umdltimls

in sheep.

MATERlALS AND METHODS

Animals:

ThfL'<: dirrcrcul sheep nocks have bceil ex.anlin(~d and a lolai of 50 shcep of bo:h sexes, ngjng

2<1 years were selected for this study, 'I11esc animals were raised freely on gmsses nod olher

field stubblcs in a new reclaimed areas and sandy soil. 111e selected animals have heen iH:}l! uu­

der dnse observation and were examined clinically aeeoruJng to Kelly {19S4). TIle exammed uui·

mals have been ~llloHed into two gOOllpS, Group -I inVOlved those clinically and apparently

healthy animals (n "" 10) and were served as control group; Mcanwlllle. ~rotiP ~2 Involved (hOHt.:

animals (n '" 40) that had the clinical sign of emaciation, rough ('oat, low n:prothtdivl' perfor'

mance. slmnling movements and poor body condltloR

Samples and sampling protocol:

Two types of blood samples have been r;:ubmttted from each animal and were subjected iiar In­

bora tory Investigations. TIle first sample was nbtaiuCt:l Oll lJeparinLzed tubes and was used imme­

diately for d('tcrmJnafJon of enzymatic acUvlties. The second sample was obtained 011 11011 heparl-

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Fouda. T. A. et Ql ... 176

nlLed lubes lor obtaining blood serum which 'WaS kept frozen untU blocheOlleal analy~ls for (he

selected p.Hmmders. 111e blood samples were obtained from the diseased as welt a~ lhe appar­

ently hc.nUhy sheep. A randomIzed milk samples were obtaIned from a number or sheep and

\\"en: Ilsed for detcnninatlon of somatic cell count as well as biochemical analysiS for some pa­

ro;;lmeters. Also, a n.:presentative samples wtrc obtained fmm the animal s diet and wen' :-ubJcd­

ed lo annlysls_

Biochemical anal,.. mood sera samples were analyzed For the selected parameters j)dl'l!r:ularly selenium by uslllP;

atomic absorption spt'f:'trophotometer. lmmunoglobulines IIlG and IgM were mcasHH'd IJY \1,,­

lop; Cd t<:teetrophoresls aceordtng to the methods adopted by Stegmann et aI. (1987),

Bioehemical analysts for determlnaUon of the enzymatic m:t1vHie~ in whole blood samples for

('utalflse enryme, superoxlde dislTlutase and CSH were carrted out ftecordlng to the mcHlOds (\{'­

wrlbcd by Cohen et al .• (1970). MIsra and FTidovich. (1972) EUmans's. (1959). rrsppdlvdy.

Meanwhile, blood seta sHOlple.$ were sU~lcdcd to the bioeh,cmlcal analysis fur de(e-rlllination of

nitr!(' oxide (Privat et al .. 1997). Wglyt"Crldes (Young and Postaner, 1975), lotal {'li()le~teml

{Oecg Hnd Zelgenohnn. 1982). high dC1H;Uy lipoprotein (Warnick et a1..1983). Low <It:lIHlly lip­

oprotein (Fdedwald et al .• 1972), serum malondlaldehydc (Daper and Hadly. 1990), '1';\ {Coop­

er,1982}, '1'4 (Schall et al*. 1978) and total lipids (Frlngs and Dunn. 1970). In uddHloll stllllHli('

{'ell eount in milk was carried out using Skar s method (A.P.H.A •• 1985).

Ration analysis:

'I1lc oblaineti samples were analyzed by wet ashing technique for detenninatioll or s('\('IJilim

cou('cnlmtinn (A.O.A.C .. 1984),

Statistical analysl$ ;

loe mCHJl values arId standard error were calculated COl" the oblalned data, ami Ihe signill­

cances for ,nil mcans have been carned out by applying Hest USing the SPSS cOllipuler pmArillll_

'flu;, \!'alw:~ have been cakulatcd aet.'Ordlng to Snedecor and Cochran (1989).

RESULTS

11le ('linie .. l examinatlon revealed Ihal. the animals of the 2nd group sbowed poor pcrJ(w-·

!Hanee witiJ the major clinIcal signs of cough, rough ('oaL iJs.5OClated with snumlng. III nddUiulI,

there Wt;f'(' a low rc.productivc perfonnancc. shuffilng movcments, dian-hea and lIJ.·thl'!I"t.

·nRc ohlHined results of biochemical analysis revealed a significant dc"Creasc in the iltl..'an val·

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Fouda. T. A. et at..

177

lies of nitric oxide {NO). lotal lipids, tnglycelidcs fTO) and T:\. Meanwhile. there is -all elevation in

the mean nllues of malomUaldhyde (MDA). eholCNlerol. low density lipoproteins (LDL). and thy­

roxin {T4) in bloou serllm of sheep with selenium dellcieney (tflblc, 11.

::\1canwhik. enzymatic adlVltles were Numma07-ed In Table (2) whll'h exhibits a signilkilnl de·

crease in tile level of catalase (CA), supcroxidc dlSrtllltasc (SOD), rcd\l{'ed ~lotalh}on(' (GSH). Clnd

glucose·6·phnsphate dehydrogenase {G6p·D)lcvcls in crythwcytt,s or sh(~cp w1th selelliulIl dr-H­

eleney.

The nblaillcd rc~ults of blood sera analysis [or selenium concl',drations and inllmlllo~tohll'

lins, particularly 19M and J:gG and cellular examination of milk ;..unples for tlll' sOlllalk cdl

eounts were expl'cssed in table (3]. The results revealed a sig:nilkuHt rcdu("UQO III iht.' menll val-­

lICS of selenium, 19.\1 and IgG io sdenium·del1etcnt sheep whclI compared \\/1111 apparently

healthy animals, associated with signilkanl ele .... ations In the lIle<lIl values of milk ,,"omalk I'd!

counts.

'nll~ obtained results of dltl analysis revealed thai scJcniwll ('oneentratlon was lowt'!' limn till'

rcrollllnen(kd requirenient for the animal: and Ule obtained vHtue wa:~ 0.056 ppm.

DISCUSSION

Sincc [he selenium intake Is eonsldcrcd to depend on the selenium content in fced HiHff;. sup­

plted to the animals; therefore it was erucial to analyze thc di.et [or !'>Clenium contnll. The ob·

tained fCSlllts of diet analysis revealed that selenium concentration was lowcr Itmll nit' IT,{",OI1l­

mended re(luirel1leni for the animals (0.056 Vs 0.1 ppmt As a dietnry level of O.l ppm sdl'nilllll

is qUil sullklent to IKevenl the signs of (kfidency in mosl animal specks (Harrison and Con­

"'d. 1984).

It was postulated by (Fang and Yang 20(2) thal, free radlcals (~upcroxide. llHrit' pXi{k Hlld

hydroxyl radicalsl and other readive spcrlcs {hydrogen peroxide, peroXYJlllritc, Hnd ltYP(lt'hlnr­

flUS a\"jell nrc produced In the body, primarily as a result of aerobic metabolism. t"mlioxi(!<m{s .', (glutathione, arginine. citrulline, 1atuine. creatine, selenium, zinc. and vltumin 1':) and Halioxi·

dant Clli)'!1lCS (superoxitlc dismutasc. catalase, glutathione reductase, and glututhlOlw p('l'()xi·

da1>;;Csl <:x('rt synergistic actions in scaven.t,.'ing free radicals and consequently proter-ting I1w body

cells alitl Ii~sues 11'001 the deleterious ciTed of free radicals,

Nitric oxide INO) Its synthesized by the enzyme nitrk" uxide synlhetase, whit-h ('\llivexis th(~

amino tn'tel L-arginine to ~Hrul1lne and nitti(', oxide. Nitric oxide plays a role In cells ,·oilnmmi­

cates wHh one another (signalll'ansdudiun), and it has n critical role in the host fCsponse In ill-

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Fouda. T. A. e:t al.~~ 178

k{"! lOlL hI I his regard, it appears that the joxle properHes of nitric uxide have been harnessed by

the immune system to kJII or at least to slow the growth of invadin~ organisms (Marletta and

Spiering. 2003).

The obtained results revealed a slgnil"icant reduction In the mean values of nitrk oxide and

glucose-6-phosphate dch}'tlrogenase In seleniunt-deficient sheep. the results which are in a!?;rce·

mcnt with [hose obtained by Litl et at. (2000) and Leopold et a1. (2003) who fountllhat. selt>

nium sollltion (~ouid increase the content of NO In the mlec'lj blood :.:cril and may bc asso('lated

with the increase in anUoxldative activity, suppressIon of (ree radk;d's intervention. and lhe ex·

eessive I>dense of NO. Oxidative stress may be induced by gcncratiOlJ of reactive oxygcn species

(ROS) and other free radicals. The generation of lliese reactive oxygen species is known \0 be m;~

soe1ated with a decrease In antioXidant levels (Yldlrlm et al .• 2003).

Leopold et aI. (2003} reported that glt1cose-6~phosphute dehydrogenase. tht~ nr~t l'm:ymr of

the pcntose phosphate pathway, Is the principal Intracellular SOurCe of NADPR NADPTI i!i Il!i~

IIzcd as a ('Of actor by vascular endolhelial edl nitric oxldc synthetase to generate Ilillic (lxitlt~

Meanwhile. It was revealed by Grasso et al. (1990) that, the production of hydrOL!cn peroxide

\tlt'r,HS('"d (wo folds In the netltrophlls obtained frOnl cows fed the selenium deJidclll did ('om~

pared w1111 ~clcl1tum st1pp!emcnlcd diet. Although hydrogen peroxide is necessary lilr mo!'>\ of Ihe

oxidatl\'e killing mechanIsms used by neu(.-ophils excess hydrogcn peroxide potclllialJy ('"nil Lw

delrtmentat 10 thc eell Itself. Increased coneentratlon of free radicals may be formed, which !'an

Inleraet with otber cellular comp<lncnts. resulting in the formation of UpJd peroxides,

Most of the mammalian selenium ts lnC<lrp<lrated or bound by certaIn proteins, sdenoprotclns

or selenium-binding proteins. respecHvety (Evenson and Sunde, 1988 and Burk et a1.. 1991].

One of the most important selenium pen!cln In mammals is ~lutathion peroxidase (Zachara.

1992), Glululhton peroxidase catalyzes the reduction of hydrogcn peroxides and Oq.!Hllil' perox­

Ides and thus protecting the membranes against oxidative damage (Ullrey. ISB7).

Selenium Hnd vitamin E act synergisticaUy and having an effect on the immune sysh'm. 'I1u .. 'y

mudUy Ihe intera(,liom' of ma('rophagcs <Jnd lymphocytes (Afzal et aI •• 1984} or Olej as 'U1lin .. '{i­

danis nn the eeUs involved in the immunological rcactions (Reffet et a1.. 1988). lymphHt'ylC5 afe

rc~arded as the cell populaUon most sensitive to pernxidatlon due to the high frcc lipid Hdd COll­

tent of their membrane,; (Nemec et aI •• 1990).

mood Icu('ocytcs have sIgnificantly hlghcr GSH .. PX activity cell than do erythrocytes (Scholz

and Hutchmson. 1979). Glutathion peroxidase may be Involved in phagocytosis by lCll('OCytc~

which is a major immune mechanism of b..'l('terial ckanmcc (Er$klne et aI •• 1987), Dtccrcu,'i('d

GS('l~PX tWllvit:v in phagocytic cell".! has been reported 1n selenium· ddlejent animals (Serfass &:

Mansour«, Vet. Moo. J. Vol. V. No. I. 2003

Toada. T. A. et at ...

179

Ganther ,1976} and has been us."ioclated with reduced bactericidal t'apaclty of I1cn!mpllils (Sex.

fass &: Ganther. 1976; Gyang et aI .• 1984; kanafani and Mattln. 1985),

Thc obtained results revealed a significant increase In '1'4 aud significant decrt'asc ill T:1 in Sf>

IClIlHm·ddki{"rH sheep. Such results cuntd he attrIbute to lhc reduction in dclodillu!o;c Iype 1 {OI­

l) aetivilics in (he periphemJ tissues tn selenium-deficIent animals (Beech et al. 1995), The re'

duellon in dclodinase uctlVJty in the peripheral tissues results in de:l:rcased rate or deiodillisallOf\

of1'4 lo'1':<: with consequent elevation of cIrt'ulaling T.j (Arthur et n!. 1990).

In this ~ludy selenhtm-dcfielent sheep showed significant redw it'nl in thc llH:an values of to~

lallipids ,and trlglycerides associated wjtb significant devalulfl-s in L1le mean V<llIlCS of cholC's\('·

rol. low dcnsHy lipoproteIn and rnalondialdihyde. Meanwhile High censlty Iipoprok'ills showrd

not) signlllx;:mt variations between the healthy and selenium deficient sheep.

'me re(I\l(~liol1 in total lipids in scIenitlm·ddkient sheep eould be attributed to tile l"t'dllC(i1l1l

in lipid absorption from small intestine due to pancrcalie u!rophy thai may Imvc ft'slIl1nl from

sdcntul'll deficiency wlUl consequent redudlon In panneatlc enzymes 11('el'$S,\I), for lipid nb:mq}­

Hon (Bunk and Combs. 19tH).

MOl"{'over. it was reported by Mazur et al. (1996} that sdcnimll deficiency have n~s!lllcd in

increased conCentration", of plasma cholcskrol and apuHpoprntein R. Meanwhilc. ~vlCllitllil ddl:-­

{'icmy in {'(lmbinafion wHh vitamIn E dcJ'idel\cjo' leads lo an increase in plasma 10\-\/ density lipo­

protein (LOLl and apolipopmtein D ('oncen! fal ions.

The obtained resnlts of this study revealed that there Is a direc! correlation between [he blond

values of selcnium and immunoglobulins, Reduced selenium concentrations In the hlond W;IS as·

socialcd with l'L"(luc-ed 19lvl and {gG in selenium-deficient sht'ep when compared with tildr \t::\'t'b

in hcallllY ones. 'Ibe results- which arc In eoncern with those reported hy Finch and Turner

(198-6). Knight and Tyznlk (1990), Larsen et ai, (1988), Mavromatis et a},(1990). and. SpaU~

holz et al.(l973}.

°nlen: is ,\Ii t'vldcn('c Indicating an enhanced irnnltll1oglobllUn M and Immuno141olmlln (; anti­

body tuns in mice fed sdenil.lm SpaUholz et aI. (1973). In addition. the resuits oblnhwd by

Flneh and Turner (1986) ~howetl that llM.rginally !:leleniull1 dcJkient lambs are able 10 die!!

strong anhbody ti[{TS to a bacterial anUgen and thaI supplemented with sdenlllm produces. HI

besl, a l1mrgrnat enhancement of the respunses obsel"\-"cd.

CllIlkally, it wali oDse.-<led in this Sll1dy that. five .-::ascs of IllHsUtlS have bcen obst'ry('d In sck­

nium-deficient sheep and associated with highly slgniHcant increase in the mean valllt~R of ~).

maUe cdl count in comparison \.\-ith the values In healthy ones, Selenium dcii<'ielH'Y has \){'t:n

assodaled with increased prevalence of mastitis- in dairy cuws und Diller luclatin~ 1llamnmls

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Fouda. T. A. et aL ••

180

(Grasso ct ai •• 1990). Goats and eatUc consuming selenIum deficient diets have milk wHh hi~h

somatic ("t:ll munts and decreased milk production (El"$kine et .Ill.. 1989), This indicates lhHI

seleniulll pli.lYs an Impmtallt role 1n mammaty resistance to infcclion, but the prt'CiHt' meelm­

nism is still unknown,

Such results cHuld be explained as, the generated peroxides leads to damage of the epithelial

cells in which llle neutrophHs have accumulated (Badwey and ka..rnovsky. 1980; Fanton and

ward. 1982). In the mammary gland, these damage 10 U1e cpHhdi;d (lssucs may lead In nit irre­

versible loss of scr.retory tissue and reduction of milk yield from U1<' :.!Iand (Grasso et at. 1990),

It could be (.'(mciudcd from thiS study that. Understanding free radicals bloluJ.tV I", lle('cs~uy

lor desi({nlng an optimal nutritional countermeasurc against cylotoxlCily. The knowlt'dg{' of cozy­

malic and non-enzyrnatjc oxidaUve defemw mechanisms wlll sCIile us a guiding print'iplc fill' es­

tabUshlnf4 Ih.: most effectIve nutrltion support 10 ensure the biological safety. Apl)wprialt' dietary

inici'VcnliollS may reduce the potentially damaging eJfccts of free mdl~als generated dllrin~ me­

tabolism and varlous physiological condlliolls. The results indIcates thal seleniuUl Cllh(lIl('('S Hit'

znUoxidanl capacity 1n oniIilllls.

Mansoura. Vet. Ned. J. Vol. V, NO.1. 2003

Foudu. T. A. et at ••

Table (J): The mean values of some blood biochemical parameters oI sch'muUl­~~~ficie11l aud clinical(v _healthy sheep"

Biochemical parameters Control animals (Grollp -1) Seleltium-dejicietll

(mimals (Group -2) Nitric oxide (nmol/ml) 48,59 ± 0,82 13.05± 063' MDA ("moVL) O.44±Om 1.41 ± 0,1' T4 (ng/ml) 35.33 ± 0,64 55.26 ± 0.61' n (ng/ml) 17,79 ± 0,92 2.38 ± 0,5S'

Total lipids (mg/dL) 427,2±21 322,6 ± 13 ' Cholesterol (rng/dL) 95.48 ± 5,9 117,64 ± 1 7' Triglyceride. (mgldL) 249,98 ±O,7 233.43 ± 1.4' HUL ( mgfdL) 45,1 ± 2.4 44,7 ± 1.1 I.UL (mgidL) 134,94 +, 46 166,28 ± 1.06"

·,Weans are sigrt~ficantly d?fferenl at the level (P _~, 05)

Table (2): The mean values of some enzymatic activities i/1 the blood of sclel/iwlI~

_ ._~/eficie1lf and clillicaily healthy sheep,

BiociJemica/ parameters COlltrol allimals (GrQup -1) Selenium-deficient

Calalase

SOU

GSH

G6P-U

(nmol/ml)

(",mollL)

(ng/rnL)

(UIIO" RBCs)

~ __ .. _____ ~ ________ , ... __ . ___ ____ ..!!..I!..n.n_i1:~s (Grou}'.. M2) 48.59 ± 0,82 13,05 ± 0.63'

1.41 ± 0, I 0.44 ± 0.07'

55,33 ± 0.64

257,) ± 5,2

35,26 ± 0,61'

210,8 ± 8,9'

·Means are significantly different at the level (P SO,05)

Table (3): The mean values of selenium and immunoglohulines in fhe blood. alUl

~1!..lilfiC cell count in the milk of selenium-deficient and clinically healthy sheep.

Biochemical potameters Control Qumtols (Group -I) Selenium~deficiellt

_____ ~~~ __ __"."imaisJGrouR.-2) .. Selenium ppm

IgG mg/dl

69,56± ),6 2156.4± 30,6

15,25 ± 2.1'

1325.5 ± 23.2'

IgM mg/d/ 196,2 ± 12,5 132.2 ± 6,8*

.S_o_m_a_li_cc~el~l~co~u~n~t _______ 7~3~,6~x~I~~~±~6~,4 ____ 2~6~7~.6~x~I~0_J±~1~2=.3=6~.4~·_ *Mcatls are slgnijicofJlly different allhe level (P ::1),05)

181

Milnsoura. Vet. Med •• J, Vol. V. No.1. 2()()3

Fouda. T. A. er: at .. 182

REFERENCES

ACzel. M.; Ttmgerdy. R.; Ellis. C. Kimbullng. C. and Morn •• C, (1984) : Protcclioll of roms

a~ain:1t epidld}'tlliUs by a brucella ovls-vit E ad,luvanl van'.Ine. Vet. Immun. Inmnmopa­

thol., 7; 293-304

A. 0, A. C. (I984) : OfUcia} methods of antilysIS. ASbocialion of oflkia.1 analytical chemIsts. 131h

edltlun. Washingwn DC. USA

A. P. H. A. (l985) : Americ.'ln Public Hcalth AssocIation, Slandard methods, 3rd edHion. Ncw

York, LISA

Arthur. J. R .• : Nieol. F.: Rae, P. W. H. and Beckett, G. J. (1990): El1cets of selenlullJ df'fit'icn"

cy on the thyroid gland and on plasma and pHtllt.o'11)' thyrulrophin and !!:I'owth horlilolle

('oll~cntraUons In the raL Clin, Chcm. EnzymoJ. COlHm, 3:209·214

Dadwey. J. A. and Karnovsky. M. L. (1980) : Active oxy,t;:Cll s-pecles and {he flllH'\iOll!' nf phuj..(o­

('ylic ICllcocy1es. Ann, Rev. Biocbem. 49: 695·726

Beech, S. G.: Walker. S. W"~ Beckett. G. J.: Arthur. J. R.; Nicol. F. and Lee. D. {1995] : 1<:1'­

feet of selenium depiction on thyroidal type-I iodolhyronlne deJodlnase adlvHy ill isola!­

cd human thyrocytcs and rat thyroid and liver. Analyst 120:827-831,

Dunk, M. J .. Combs~ G. F. Jr. (1981) : lkJntionshlp of selellium-dependent gluiailliOlIC pcruxi.

di.\sc activlty and nutritional pancreatic atrophy in sclcnium-ddklclil ('l!kk.,,>. J

NI)lr.: 111(9): H31 1-20

lJurk. R. F.: hill. H. E.; Read. D. R. and Bellew. T. (1991) : l<:esptJnsc of ral se\('lIoprulcin l'

io selenium adminIstration and iaie of Its selenium. Am. J. Physiol. 261. E2G-I~;m

Cohen. G. Oembiee. O. and Marcus, J. (1970) : Measurement of catalase acHvlty in tissue

(·xtnwls. Anal, Bioch.34:30-38,

Combs. G. F. and Gray. W. P. [1998): Chemopreventive agents: SeJenium. PharnmeoloJ!..vamI

·l1l!:rapcutics. VoL 79; 197-192

Cooper, E. A" (198!tl) ; Radioimmunous.<;ay of tllyroxinc and 3,3,5 tr11odothyronirw {rrvCl'se) ill

humun amninlW fluid. Clin. ChC»l. Ada, llA,57~66.

naper. H. H. and Madly. M. (1990) : Malondlaldhydc determination as index of lipld I1t~roxida~

lion, Methods gnzymoL186:421-431.

Debskl. D .• Zarski. T. p, and Milner. J. A. (1992) : Selenium contcnt and d1strlbutltm in bovlnc

nn human milk from dl.llerent regIons of Poland, J. VeL M('d.A. 39: 313-318.

Deeg. R. and Zelgenohrm., J, (198.2) : Enzymatic and eolorlmcteric t('st for dctcnnil1Hliol1 nl' LO'

MartSOuru. Vet. Med. J. Vol. V. No.1. 2003

Fouda. T. A. €t 111. ••

183

tal eholcsteroL CHn. Chem.28, 1574.

Ellman's, G.L. (1959) : Tissue sulphahydryl groups, Arch, lliochem.

Erskine, R. J.: Ebel'hart. R. J.; Hutchinson. L, J, and Schou, R, W. (989) : HiOlXI

sc!enJum conccntrat1ons ulld ~ll\lalhtone peroxidase u('UviUes in dairy herds with

high and low somatic cell coun/s, JAVMA. Vol. 190. No. I I.

Evenson, J. K. and Sunde. R, (1988) : -Selenium inCOl"pora!1oll illll! sclennproleins In lhe St,

adequnle and Se deficient rat. Pro('. Soc. Exp, BioI, :\100 .. ! "~'I': 169-180

Fang. Y. Z., Yang S. and. Wu,G. (2002): Frce radicals. anlim..id;nn". nnd I1lllrilimL Nu!riiioll.

Od; Jail 0):812·9.

Fantone, J. C. and. Ward, P. A. ~1982} : Rotc of oxygen deriVed freT radicals and ml'i.;\!Jolt!t'S In

lcu('ocyte-dependenllnflammatory reactions. ;\Ill, J, PathoL 107: 397-418

Finch. J. M, and Ttttner, R. J, (1986) : SelenIum suppkmcnlallon in tam!)',: dkc[s nil

antibody responses to a salmonella vaccine. Vet Ret'. Oct 25: 119; 171:4~m- ! ,

Fliedwald. W. T.: Levy. R. [. and Fl'edrlckkson. D. S.(1984) : EsHnlHUnn of LDL-c in plasma

wit hOll! ll$e of prcpruUve ultraccntrifuge, Clin.Chern, 18.49~·502,

Fringe:. C. S. and Dun.n. R. J .• (1970) : Colorirncilic dctcmlinatiol1 or :it'I'lIIll j1!\a! lipid", lJil.'wcl

Oll sl1fpho~ph(!sphovanillln reaction. Am, J ,CHn. Palho1.53, H9- 9 1.

Gates, N, L. and. Johnson. K. A. (2002) : Sclt'nlum related disordcr~ in W.tshlll!!!()l1 Uvcshwk.

htl 1':/ ! cru.cache,WSH.cdu/Cc:publications/cb 1607/eb 1607.hlml

Grasso. P. J.; Scholz. R. W.; Erskine. R. J, and Eberhart, R. J. (1990) : Phagocylosb. bacted­

cidal activity, and o.ddaUve met.-lbollsm of milkncu\lophils hum dalry ('ows fed sdt'ni ..

um supplemented and selenium delklcnt diets. Am, J. Vet. Hes .. Vol, 51, No, 2: 269

Gyang. E. 0,; Stevens, J. B, and Olson. W, G. (1984) : EJkcts of selenium vitumin E Ini('('llt){);S

on bovine polymorphnueleated iencocyes phagocytosis and kiUin of Staphylili'O('('IIS au­

re\ls. Am,), Vet. Res .. 45: 175·]77

Hamson. J. H. and Conrad. H. R. (l984) : Selenium content and glutathione pcrnxidafw,l('­

IIvHy jn tissue. J. Dairy Sci., 6-7: 2464·2470

Holben. D. H, and Smith. A. M. (1999) : The diverse role of the sdenltllH within

~lenoprotejl1s: A rcview. JOllrthl1 of the AmeriC'{Ul Dietetic A%otiatinn. Vol. 9!):

H:}fi-84~{

Kana(ani. H. and Martin, S, :E. (1985) : Ca!<\lse and Stll:IC1'oxide disl1luiasc udiviUct-; ill virnk'l!l

ilml non~virulent Staphylococcus allrellS Isolate-s. J. elin. MlcrobloL, 21: 607·G10

'Mansouro. Vet. Med, J. Vol. v. No.1. 2003

184

Kelly. W. R. (1984) : Vetertnary Cllnieal Diagnosis. 3rd edition. BuiUlar Tindall. Londun

Knight, D. A, and Tyznik. W, J. ~1990) : TIle .. Heet of dietmy selenium nn hl.mlOral

iillnlUnoCQmpetcnee of pollics. J ArHm Sci. May:68(51: 131 1-7

Larsen. p, R. (1998) : The thyroid gland. In Wilson ct at Edltion, Williams Textbook nf Endo­

crinology, 9th Lxlttiotl. 389~515. WJ3, Saunders. PhIladelphia

Lstrsen. H. J.; Moksnes, K. and OVemes, G. (1988) : lnlluen. {' of selenium on antibody

production lu sht--ep. Res Vet Sr.I du1;45t 11:4-1 0

Leopold. J. A.; Walker, J.; Scribner, A. W.; Voetsch B.: Zhang. Y. Y.; Loscalzo. A. J.; Stan­

ton, R. C. and Loscalzo. J. (2003) : GltH'osc-6-phosphntc dehydrop:en(lst' lHndulates

vascular endothelJal growth fU<:toT-mcdialed angiogenesis. nioL Chem.30

Llu. S.; ShiH, 0.; Uu. G.; Chen. H.; Liu. S. and Hu, Y. (2000) : Rolc$ or Se and NO in apoplO .. ';ts

uf hepatoma cl'-lIs. Life ScL29; 68 (61 ; 603-10.

Marietta. A. and Spiering. M. (2003] : Trace elements and nitri(! oxide funC'linn, d. NUT. !3:J

151,1431J433,

Mav:romatis, J.; Koptopoulos. G.; Kyrlak18. S. C.: Papasteriadis, A. and SaouJldts. K.

(1999) ; ElTeets of lo('opherul amI "dc.nium on pregnant sows and their piglcls huum·

nity nod perionnan('c .• J. VeL Mcd" A. 46, 545-553

MUllr, A.: Nassir. Jr.: Gueux. E.; Maundras C.; Bellange:r. J.; G:rolier. P.; Rock. E •. and Rays­

slguIer. Y. (1996) : Dicts dcficlent in selenium and vitamin E alTc('t plasma tipopro4dn

and apollpoprotcin cou{''entrations In the rat. ilL J. Nutr,76i6):S99·907,

Misra. H. P. and Frldovich. I. (1972) : Thc role of superoxide dismutase uoion In tlw oxidatlnn

of epinephrIne and simple assay lor superox1dc disHlutasc, J, Blot. Chcm. 247:3170.

Nemec. M.; Hidiroloou. M.: Nielsen. K. and Proulx, J. (1990) : Effeet Orvitamin f':;' .. ml sdclli­

um supp1cmentJl,Uon on some immune parameters follo'W1ng vaccination again",! bn.wcl­

)(lsis 1n cattle, .J. Animal Sci.. 68: 4303~4;309

Privat. C.; Lantolne. F.: BedJoul, F.; Van Brussel. E. M.; Devynck. J. and Oevynek. M. A.

[1997) : :.litrle oxide prOOl,.leHQtl by endothelial ('~II~: eomparlsoD of three n)('lhod~ of

quantification .. Life Sci 61: 119:]-1202,

Reffett, J. K.; Spears, W. J. and Brown, T. T. ,1988) : Effects of dietary selenium on lite pri·

mary and secondary immune response In calves clml1cngcd with infectious bovine rhi­

llo(meheHis virus. J. NutoUQo. 118; 229-235.

MUMouro. Vet. Meet J. Vol. V. NO.1. 2003

Fouda. T. A. et aL.

185

Schall. R. F, ; Faster, A. S.; Hansen. H. W.; Kern. C. W. and Tenogo, H, J. (1978) : A scm;j·

tiw manual enzymc imml1ll0u"""tlY lor fhyroxine. Clin. ChclIl.24. 1801- IH04.

Scholz. R. W. and Hutchinson. L. J. (1979) : DistribUU(!fl of glllLalhlone perPxiddsl' adivity

and selcnium in the blood of daily cows. Am. J. Vet. He:'> .. 40: 245 .. 249

Senedecor. G. W. and Cochran. p, W. (1989) : Statistical Mt:thmls, Bth cd" lnw;,~ 51<\11' Univer­

sHy. Press. Nnes, LA.

Serfass. R. F. and Ganther. H. E. (1976) i Effeets of dlCb.ny sekl·jllm and tOl'oph('wl un gIll'

ldthione pe-coxidasc and superoxidc dismutase activities ill r t! phagocyirs. Life Set. 19:

1139-1144.

Stegmann, H.; Burgermeistel'. H. and Klogeneeb Lenfort. E. (I987) : Pcnla-phnn-' <:Ind

fllOllo-phnrc ekctrophoresis and isoeleetric ro(:usin~. lsi edilion Wafer lit' C,myler.

fkrllfl

SpaUholz. J. E.; Martin. J. L.; Gerlach, M. L. and Heinzerllng, R. H. (I973) : Enlimw('d im­

mUlloglobulin M and JnllnunoglnbuIin G tlnlibody titers In mice fed seleilium. tnlcvl

tlillnun. Nov:8(5):84I w2,

UUrey. D. E. (987) : nCocheml('al and physiolo~it'al indications or selenium "lalus In

.anlmals. J, AnimaJ Sci" 65: 1712-1726,

Warnick. G. R.; Benderson, V. and Albers. N. {1983} : Selected mcthod for tldel'lHinaliol\ of

HDL-c. Clio. Chcm.1O:91-99.

Weisburger. J. H. (l99S) : Eva1uallon or rhe evidence on the rnk of lol1lalo prodtWls ill disca.:x·

prcvenlion. Prot:. Soc. EXp. DioL Mcd.;218(2): 140-14:1.

Whanger. P. D.; Weswtg. P. H. and Schmitz, J. A, (I977) : l-:Hed~ of selenium and vitamin I~

011 blood selenium levels, glutathione pcroxidase actlVlties and while mllsdl' tlise<lsC in

sheep fed purmed or hay d1ets. J, Nutrition. 107: 1298·1307

Yildirim. M,; Baysai. V.: lnatOI. H. S.; Keslci. D. and Delibas. N. [20(3) : 'I1le role of oxidan!s

tind antioxidants in generalized vHlligo. J. DennatoL:30{2J; 104·8.

YOtUlg, D. and Postaner. L. (1975); Eillymalic and {'olorilllf:!('ck le::-t for detenlllH,,\il)lI of Irl­

J,!ly<'cridcs. ClirL Chem.21 .5,

Zachara. B. A. (1992) ~ Mammtlian sclcnoprntei:ns. J. Traf'e Element Electrt)lyj(:~ !k"llh Dis­

caHC.6: 137·151.

Mansoura. Vet. Med. J. Vol. v. No.1. 2003

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