Effect of vitamin E and selenium supplement in reducing aflatoxicosis on performance and blood parameters in broiler chicks

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1 Irqi Journl of Veterinry Sienes, Vol. 23, Supplement I, 29 (97-13) Proeedings of the 5 th Sientifi Conferene, College of Veterinry Mediine, University of Mosul Effet of vitmin E nd selenium supplement in reduing fltoxiosis on performne nd lood prmeters in roiler hiks A. A. Shlig College of Agriulture, Tikrit University, Slh Al-deen, Irq Astrt This study ws imed to investigte the suffiieny of Vitmin E nd Selenium Supplementtion to Diets ginst ontining fltoxins on the reltive orgn weights nd vrious struturl lood prmeters. One hundred nd twenty unsexed Ross irds were used from 3 to 7 weeks of ge. Birds were rndomly distriuted nd sujeted to five nutrition tretments s follows: (T1. Control Group:. AF +. Se +. Vit. E, T2. mg AF/kg diet, T3. mg AF /kg diet +.18mg/kg Se +1 I.U Vit. E, T4. mg AF/kg diet +.32mg/kg Se +3 I.U Vit. E, T5. mg AF /kg diet +.5mg/kg Se +5 I.U Vit. E) pollution y fltoxins uses signifint inrese in reltive weights of liver, hert, gizzrd, domen ft, nd spleen. A signifint derese in totl ody nd rss weight gstrointestinl trt long. Moreover existene of fltoxins used signifint derese in vlues of Pked ell volume (PCV), Red lood ells ounts (RBC), Hemogloin (H), men orpusulr hemogloin (MCH) men orpusulr hemogloin onentrtion (MCHC) signifintly moreover, it hs een notied signifint inrese ourrene in vlues of totl white lood ell (WBC) nd rtion of Heterophills to lymphoytes in seond tretment irds influened y Afltoxins. The ddition of grded levels of Vit. E nd Se to the AF ontining diets (T4, T5) improvement totl ody nd rss weight nd internl orgns studied. The ddition of oth Vit. E nd Selenium (T3, T4, T5) to the provender ontining y fltoxins, hd no effets on lood prmeters to their nturl verges t ontrol group. The existene of the two Vit. E nd Selenium in polluted provender doesn't prevent derese in these vlues with signifint improvement (up to 1 I.U Vit. E nd.18mg/kg Se) ourrene from irds group vlues whih hd een fed y polluted provender only. Keywords: Vitmin E, Selenium, Afltoxins, Broiler. Aville online t E E. Ross ( ) :T1). ( ) ( ) ( ) :T4 /.18 +E I.U 1 + / :T3 / :T2 I.U 5 + / :T5 /.32+ E I.U 3 + /.( /.5 + E (P<, ) / 97

2 Irqi Journl of Veterinry Sienes, Vol. 23, Supplement I, 29 (97-13) Proeedings of the 5 th Sientifi Conferene, College of Veterinry Mediine, University of Mosul ( ) (T4,T5).. Interodution Afltoxins (AF) re seondry fungl metolites produed y some strins of Aspergillus flvus nd Aspergillus prsitius. Afltoxin B1, B2, G1 nd G2 re nturl ontminnts of mny niml feed ingredients. AFB1 is the most undnt nd toxi form of ll nturlly ourring fltoxins lso represents 75 of ll fltoxins found in ontminted food nd feeds. It is heptotoxi (1), hep-torinogeni (2), nd tertogeni (3) to vrious niml speies. They were lssified s group I rinogen in humns (4) Afltoxin B1 is iotrnsformed in the liver y monoxygenses nd then trnsformed y ytohromo P45 into fltoxin 8-9 epoxide nd hydroxylted metolites (AFM1, AFP1, AFQ1 nd fltoxiol) (5). Both Vitmin E nd Selenium re essentil nutrients for humns nd nimls. They re involved in the protetion of iologil memrnes ginst lipid peroxidtion nd preventing the free rdils dmge to phospholipids memrnes nd enzymes (6). Vitmin E is the mjor lipid solule ntioxidnt tht is present in iomemrnes, svenges free rdils in the erly stges of lipid peroxidtion nd Selenium is essentil to the tivity of glutthione peroxidse (GSH Px) whih redues H 2 O 2 nd lipid hydroxides to less retive produts (7). It is well understood tht Selenium hs strong intertion with vitmin E in the protetion of ell from importnt moleules (6). Alph toopherol nd glutthione peroxide protet tissues from oxidtive dmge ssoited with the free rdils generted during the respirtory urst of mrophges nd neutrophils (experiened during immune response) (8) nd improvement the ird helth nd immune sttus, Vitmin E nd Se re ntioxidnts essentil for ell survivl in environments ontining peroxides (9,1) The response of roilers to Se supplementtion ould vry gretly with the level of Vit. E in the diet (6) Selenium is etween 5 nd 1 times more effetive s n ntioxidnt thn vitmin E. while Se supplementtion rte of.1-.5 mg/kg is stisftory for most nimls inluding poultry (11) Vitmin E ws reported s n exellent iologil hin reking ntioxidnt tht protets ell nd tissue from lipoperoxidtive dmge indued y free rdils (12,13). Chikens, however n not synthesize Vit. E suh requirements must e met from dietry soures (14) Supplementtion with 1 I.U of Vit. E /kg feed hs een shown to derese the onentrtion of mlondildehyde produt of lipid peroxidtion in the livers of hikens fed ohrtoxinin A nd T-2 toxin ompred to dietry level of 1 I.U of Vit. E /kg feed (15). A highly tive eletrophili ompound tht is intivted y onjugtion with glutthione s- trnsferse (GST) to the AFB glutthione nd exreted through urine nd ile (16). High onentrtion of fltoxin B 1 -GSH-Px onjugte nontoxi metolite ws produed in ile id from rts fed diets ontining fltoxin with Se. Thus dietry ddition of Vit. E nd Se my llevite the fltoxiosis y inresing GSH- Px tivity from IU/g H to186.2 IU/g H nd eventul hnges of toxi sustnes to inert metolites (6) The ojetives of the present study ws to evlute the effet of dietry fltoxin ontmintion lone or with the ddition of Vitmin E nd Se on roiler performne nd lood prmeters. Mterils nd methods The experiment ws onduted t the poultry prodution frm, College of Agriulture, Tikrit University during the period from 4th Mrh to2nd April -29. The totl numer irds ws 12 twenty one dy-old Ross unsexed hiks were individully weighed, wing nded, nd housed in 15 pens under ontinuous lighting. Chiks divided t rndom into 5 tretment groups. Eh one ontins 3 replites. Eight roiler hiks for eh replite group were fed orn soyen mel sed finisher diet ws otined from it ontined 18.3 rude protein, 1.3 lysine nd.52methionine, 31 Kl /Kg metlizle energy. The feed nd wter were d-liitum.chiks were rndomly ssigned to the following tretment groups (T1. Control group: Corn soyen sl diet+. AF +. Se 98

3 Irqi Journl of Veterinry Sienes, Vol. 23, Supplement I, 29 (97-13) Proeedings of the 5 th Sientifi Conferene, College of Veterinry Mediine, University of Mosul +. Vit. E, T2: sl diet+ mg AF/kg diet, T3: sl diet+ mg AF /kg diet +.18mg/kg Se +1 IU Vit. E, T4: sl diet+ mg AF/kg diet +.32mg/kg Se +3 IU Vit. E, T5: sl diet+ mg AF /kgdiet+.5mg/kgse+5i.uvit. E) (Se nd Vit. E were Otined from the stte ompny for drugs industry nd medil pplines, Smrr). Afltoxin (AF) ws prepred through inoultion of rie y Aspergillus prsitius NRRL 2999 (generously from the ollege of griulture, Tikrit Univ., Slh Aldeen, Irq) s desried y (17) nd modified y (18). Fermented rie ws then utolved nd ground, Afltoxin onentrtion ws mesured y spetrophotmetri nlysis (19,2) Lter the rie powder ws then mixed with the diet to otin the level of mg AF/kg. Six irds from eh tretment groups were led y rdi punture for determintion of lood piture. The totl erythroyte ount ws mesured ording to (21). Hemogloin ws determined y the method of (22) Hemtorits were mesured y the miro- hemtorits entrifugtion method while the men orpusulr volume (MCV) men orpusulr hemogloin (MCH) nd men orpusulr hemogloin onentrtion (MCHC) were lulted (23) the sme irds were then killed y ervil dislotion Adomen ft, liver, kidneys, spleen, proventriulus, gizzrd nd hert were refully removed to determine their reltive weights nd dressing perentge of rsses. Dt for ll response vriles were sujeted to ANOVA. The results otined were sttistilly nlyzed using (24). Men showing signifint differenes were ompred y multiple rnge test (25). were onsidered signifint sed on the.5 level of proility. Results The effet of AF lone nd when omintion with grded ddition of Vitmin E nd Se on performne of growing hiks nd the reltive weight of internl orgns from 21 dy old to the ge of 46 dy old show in tle (1). Chiks fed the diet ontining mg AF/kg hd signifintly (P<.5) lower totl ody nd rss weight with length of the gstrointestinl trt. The results indited tht the presene of AF lone in the diet used signifint enlrgement in the size of internl orgns like liver, kidney, spleen, gizzrd, hert nd domen ft. These findings greed with those otined previously (26-28) the liver is onsidered the priniple trget orgn for AF it ws evident from the dt of the present study tht feeding the diet ontining AF lone resulted in signifint inrese in the reltive weight of liver s well s other internl orgns inluding hert kidneys gizzrds proventriulus spleen nd pnres. The results of third tretment differ signifintly from tht of the ontrol group totl ody, rss weight nd gstrointestinl trt long with signifint (P<.5) inresed reltive weights internl orgns. The grded ddition of Vit. E nd Se t the two stted levels T4 nd T5 meliorted prmeters mentioned ove in omprison those of the ontrol group or those of irds fed the diet ontining AF lone. It is thought tht the ddition of Vit. E nd Se (T4,T5) to the AF ontining diets helped in restoring the reltive weights of liver, kidney, spleen, proventriulus, gizzrd nd hert lose to those of the ontrol group. No signifint differenes were oserved in reltive weight of the ove mentioned orgns etween those of the ontrol group nd those fed the AF ontining diets mended with.32mg/kg Se +3 IU Vit. E nd.5 mg/kgse+5 IU Vit. E. The toxin fed irds treted with vitmin E nd Se showed signifint gin of ody weight s ompred to toxin lone fed irds inditing the enefiil effet of vitmin E nd Se on totl ody weight gin during fltoxiosis. The effet of fltoxin with omintion with Vitmin E nd Se on seleted lood prmeters re presented in tle (2). Vlues of hemogloin, hemtorit, men orpusulr volume, men orpusulr hemogloin, nd men orpusulr hemogloin onentrtion were signifintly (P<.5) deresed in hiks fed the diet ontining AF lone. Blood piture of tretment three ws signifintly deresed in H, RBC, PCV, MCH nd MCHC vlue when ompred with ontrol group. Exept MCV vlue whih no signifint differenes were oserved these findings re in greement with those otined previous (29,3). The ddition of Vit. E nd Se t the two stted levels (T4 nd T5) ws similrly effetive in lleviting the negtive effet of AF on ll lood prmeters H, RBC, PCV, MCH nd MCHC signifintly improved the vlues of when ompred with those of irds fed the diet ontining AF lone ut they were still signifintly lower then those of the ontrol group. Dt presented in tle (3) show the effet of dietry tretment on the results indited tht the presenes of Afltoxin lone in the diet used signifint (P<.5) inrese in leukoytes, heterophils nd H/L rtio ompred with those of irds from the ontrol group. with signifint (P<.5) derese in monoytes, lymphoytes, sophile nd eosinophi sme effet shown in tretment three irds result exept eosinophi vlue These findings greed with those otined previous (31,32). The ddition of Vit. E nd Se to the diet hd signifint (P<.5) effet on studied (T4 nd T5) with those of ontrol group. Furthermore levels of irds fed the AF ontining diet mended with Se nd Vit. E However lthough the ddition of Vit. E nd Se to the AF ontminted diets signifintly improved the vlues of when ompred with those of irds fed the diet ontining AF lone ut they were still signifintly lower thn those of the ontrol group. The results of this study suggested tht the Vit. E nd Se in omintion n prtilly meliorte the toxiity of AF in roilers. 99

4 Irqi Journl of Veterinry Sienes, Vol. 23, Supplement I, 29 (97-13) Proeedings of the 5 th Sientifi Conferene, College of Veterinry Mediine, University of Mosul Tle (1) Effet of grded ddition of Vitmin E nd Se on reltive orgn weights of hiks fed diets ontining mg AF/kg diet. Tretments AF mg/kg Vit.E IU Se mg/kg Finl ody weight Crss weight Gstro-intestinl trt Proventriulus Reltive orgn weight (g/1 ody weight) Gizzrd Liver Pnres Spleen Hert domen ft dressing perentge T T T T T Mens within olumn with different letters differ signifintly (P<.5) Disussion There re mny reports on the effets of vrious foods or nutrients nd xenoiotis on AFB1-mromoleule dduts formtion. Oviously the mjor ojetives of these studies were to determine if nd how those nutrients or xenoiotis ould ffet dduts formtion espeilly DNA ddut. This study ws undertken to evluted the effet of feeding AF ontining diet mended with Vit. E nd Se.Inresing Vit. E level in the diet from minimum requirement reommended y (33) 1 IU to 5 IU kg/ diet nd Se from.18 to.5 mg/kg diet influene importnt prodution prmeters suh s performne of growing hiks, reltive weight of internl orgns, hemogloin, hemtorit, men orpusulr volume, men orpusulr hemogloin, nd the men orpusulr hemogloin onentrtion. (34) reported tht the omined Se (.2 mg/kg s N2 SeO3)-VE (1 IU/kg) defiieny enhnes tivtion or inhiits detoxifition of fltoxin B 1 1mg/kg (deresed AFB 1 inding to DNA, RNA, nd protein) in white leghorn hiks whih ws ompletely effetive in preventing oxidtive dithesis nd deth. (35,36) reported tht high onentrtion of fltoxin B1-GSH Px onjugte non toxi metolite ws produed in ile id from rts red diets ontining fltoxin with Se. Thus dietry ddition of Se my llevite the fltoxiosis y iresing GSH Px tivity nd eventul hnges of toxi sustnes to inert metolites. They lso reported tht the Vit. E redued the formtion of AFB 1 dduts in the liver. The oservtion tht the rts fed AF free diet ontining 12 IU VitE/kg showed signifintly enhned lood GSH Px The feeding of AF diets ontining 6 IU Vit. E/kg indued signifint derese in the liver lipid ontents (37). 1

5 Irqi Journl of Veterinry Sienes, Vol. 23, Supplement I, 29 (97-13) Proeedings of the 5 th Sientifi Conferene, College of Veterinry Mediine, University of Mosul Tle (2) Effet of grded ddition of Vitmin E nd Se on hemtologil vlues of hiks fed diets ontining mg AF/kg diet. Tretments AF mg/kg Vit.E IU Se mg/kg Hemogloi n g/dl R.B.C (1 6 / mm 3 ) P.C.V MCV (µm 3) MCH pg MCHC g/dl T T2 T T T5 5.5 Mens within olumn with different letters differ signifintly (P<.5) Tle (3) Effet of grded ddition of Vitmin E nd Se on Differentil ount of norml leukoytes hiks fed diets ontining mg AF/kg diet. Tretments AF mg/kg Vit.E IU Se mg/kg Leukoytes (1 6 / mm 3 ) Monoytes Lymphoytes Heterophil Bsophile Eosinophil H/ L rtio T1 T2 T3 T4 T Mens within olumn with different letters differ signifintly (P<.5)

6 Irqi Journl of Veterinry Sienes, Vol. 23, Supplement I, 29 (97-13) Proeedings of the 5 th Sientifi Conferene, College of Veterinry Mediine, University of Mosul The inhiition of ody growth nd the deline in rss musles weight, musle protein nd RNA synthesis pper to e solely the result of the profound effet of AF. Inhiition of ellulr proesses the most prevlent symptom of fltoxiosis in poultry is redued growth rte nd poor performne. These dverse effets of AF due to norexi, listlessness, inhiition protein nd DNA synthesis nd lipogenesis (38) AF ppers to exert its negtive effet on irds hiefly y depressing DNA nd RNA synthesis nd protein synthesis whih eventully refleted in the finl ody weight nd lter rss yields (49). (4) find out the influenes of 2 mg/kg of fltoxin B 1 nd immunomodultors on the performne of roiler, fltoxin B 1 lone hd signifintly redued the ody weight feed effiieny rss yield immune development ginst Newstle disese in roilers supplementtion of ltoilli, Vit. E nd Se hd improved the ody weight, feed effiieny, rss yield nd immune ginst Newstle disese. Feeding of fltoxin B 1 (1 mg/kg) to 2- week old Jpnese quil for period of 8 weeks inditing tht supplementtion of selenium selentie 5 mg/kg hd some protetive tion ginst the toxi effet mirosopi hnges in the liver hert nd urs of friius lymphoid ggregtion in liver fltoxin B 1 (41). An improved ntioxidnt sttus ws oserved in hiks of seond group with α-toopherol nd selenium supplementtion inluding higher onentrtion vitmin E, inresed tivity of glutthione peroxidse nd lower levels of lipid peroxidtion (42). Aknowledgments This study ws supported y the College of Agriulture, Tikrit University. nd the stte ompny for drugs industry nd medil pplines, Smrr. Referenes 1. O Brien K, Moss E, Judh K & Nel G. Metoli sis of the speies differene to fltoxin B1 indued heptotoxiity. Biohem Biophy Res Commun : Admson RH, Corre P, Seier, SM, MIntire, K & Dlgrd D.W. 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7 Irqi Journl of Veterinry Sienes, Vol. 23, Supplement I, 29 (97-13) Proeedings of the 5 th Sientifi Conferene, College of Veterinry Mediine, University of Mosul 34. Chen J, Goethius M.P, Cmpell T C nd Coms G F Jr. Effets of dietry selenium nd vitmin E on hepti mixed-funtion oxidse tivities nd in vivo ovlent inding of fltoxin B1 in rts. J Nutr.1982.; 2: 324: Degn,G.H. nd H.G. Newmn. The mjor metlites of fltoxin B1in the rt is glutthinon onjygte.chem.biol.inttt.1978;22: Weiss W P, Hogn J S, Smith K L, Hoiet K H. Reltionship mong selenium, vitmin E, nd mmmry glnd helth in ommeril diry herds. J Diry Si.199;73: Choi,Y.K.,K.K.J.jung,K.Y.Che,I.Jng,B.D.Lee.nd K.H.Nhm..Effets of vitmin E nd Selenium supplementtion to diet ontining fltoxin B1on the ontents of liver lipids nd vrious lood prmeters in rts. AJAS.1995;8 : Oguz H., Kurtogh V. Effet of linoptilolite on fttening performne roiler hikens during experimentl fltoxinosis. Brit Poult Si.. 2; 4: Afzl N, Devegowd. G D. The effet of grded levels of dietry fltoxin on ertin iohemil prmeters in roiler reeders.wpc 24 XXII World s poultry Congress, Istnul, Turkey Mni,K K, Sundresn nd Viswnthn K. Effet of immunomodultors on the Performne of roilers in fltoxiosis. Indin Vet J 21; 78: Jkhr K K, Sdn J R. Sequentil pthology of experimentl fltoxiosis in quil nd the effet of Selenium supplementtion in modifying the disese proess. Myopthologi. 24;157(1): Sodhi, S. A. Shrm A.P S. Brr R. S. Effet of α toopherol nd Selenium on ntioxidnt sttus, lipid peroxidtion nd heptopthy indued y mlthion in hiks. pestiide Biohemistry nd physiology.27; 9: