Immune Response of Broiler Chicks Fed Yeast Derived Mannan Oligosaccharides and Humate Against Newcastle Disease

Transcription

1 World Applied Sciences Journl 18 (6): , 2012 ISSN IDOSI Publictions, 2012 DOI: /idosi.wsj Immune Response of Broiler Chicks Fed Yest Derived Mnnn Oligoscchrides nd Humte Aginst Newcstle Disese 1 2 Abdolhfm Mehdi nd Ghhri Hsn 1 Fculty of Veterinry Medicine, Urmi Brnch, Islmic Azd University, Urmi, Irn 2 Deprtment of Poultry Science, Fculty of Veterinry Medicine, Urmi Brnch, Islmic Azd University, Urmi, Irn Abstrct: The im of study ws to investigte e effect of yest cell wll derived mnnn oligoscchrides (MOS) nd Humte (HU) supplemented to broiler diet on ntibody response ginst Newcstle disese virus (NDv). The investigtions were crried out in completely rndomized experimentl design. Chicks were divided into 8 tretment groups, wi 4 replictes per tretment nd 10 chicks per replicte. Tretments were: 1) negtive control group, neier vccinted ginst Newcstle disese virus nor given dditives; 2) positive control or broilers were vccinted ginst Newcstle disese virus + 0 dditives; 3,4 nd 5) positive control plus diet supplemented wi 0.1, 0.2 nd 0.3% humte, respectively; 6, 7 nd 8) positive control plus diet supplemented wi 0.1, 0.2 nd 0.3% mnnn oligoscchrides, respectively. Broiler ws fed corn-soyben mel bsed diets. For ntibody nlyses, blood smples were collected by wing veins nd e titers of ntibody ginst Newcstle disese virus were determined using hemgglutintion-inhibition test. Compred to e positive control, e ntibody titers ginst NDv were determined significntly lower in negtive control group from 35 to 42 dys of ge. This study demonstrted n increse in e ntibody titers in broilers fed diets contining HU nd lso indicte t inclusion of MOS resulted in incresed ntibody titers ginst NDv in e ird, fif nd six weeks of ge. Thus immune function could be modified wi dietry mnnn oligoscchrides nd humte supplementtion. Key words: Mnnn oligoscchride Humte Humorl immunity Newcstel disese Broiler INTRODUCTION trnsglctooligoscchride nd mnnn oligoscchride (MOS). The MOS is derived from e outer cell wll of The worldwide increse in e use of ntibiotics s n yest nd its evlution in diets for breeders is of integrl prt of e poultry nd livestock production prticulr interest becuse it not only shifts industry to tret nd prevent infectious bcteril diseses gstrointestinl microflor blnce towrd beneficil nd s grow promoters t suberpeutic levels in feeds orgnisms [1] but lso hs immunomodultory properties hs led to e problem of e development of bcteril [2]. Scchromyces cerevisie nd Scchromyces ntibiotic resistnce during e pst yers. Alterntive boulrdii re e exmples of yest lso used in solutions to e use of ntibiotics hve been sought nd preprtion. The yest cell wll hs powerful ntigenic ere re now vilble number of direct-fed microbil stimulting properties nd it is well estblished t or probiotic products which help to mintin e blnce is property is chrcteristic of e mnnn chin [3]. of e intestinl microflor in rnge of food niml Supplementtion of poultry diets wi MOS results in species. Severl pproches re to use Probiotics, improved production in terms of body weight gin prebiotics, orgnic cids nd enzymes/microbil to nd feed conversion [4], prtly due to its hypoesized competitively exclude pogenic bcteri from e GI nutrient spring effect but primrily due to its influence trct of livestock; using well-defined gents t tckle on nutrient utiliztion in e gstrointestinl trct [5]. In bo endemic disese nd food borne zoonoses. Prebiotic ddition, previous reports suggest t MOS includes fructooligoscchride, glctoligoscchride, supplementtion resulted in significnt improvement in Corresponding Auor: Ghhri Hsn, Deprtment of Poultry Science, Fculty of Veterinry Medicine, Urmi Brnch, Islmic Azd University, Urmi, Irn. 779

2 ntibody responses in broiler nd lyers [2]. Beneficil Antibody responses hve been used s mesures of e effects of yest products in ruminnts re due to humorl immune sttus of bird. From is perspective; one incresed concentrtion of totl nd cellulolytic ruminl of e objectives of our study ws to investigte e bcteri [6], which my increse vilbility of ME from effects of feeding MOS-supplemented diets to broiler on diets, ereby incresing production. Some studies hve Antibody titers ginst Newcstle disese virus. confirmed e effects of yest culture (YC) in incresing Humte is highly compressed prehistoric compost concentrtions of commensl microbes or suppressing t cn be mined from deposits in e ground. Humte is pogenic bcteri [7]. In recent in vitro study [8], e composed of minly humic cid, crbon nd some orgnic ddition of soluble frction of YC showed n mtter. Humic substnces re widely spred in nture. ntiinflmmtory effect in conjunction wi ctivtion of They occur minly in hevily degrded pet but lso in ll nturl killer cells nd B lymphocytes. In ddition, oers nturl envirornnents in which orgnic mterils nd hve reported t yest products ffect nutrient microorgnisms re, or hve been present [14]. The digestibility [9] nd intestinl mucosl development [10]. romtic structures re ubiquitous in humic substnces. Furermore, supplementtion wi oligoscchrides my Specil physicochemicl property of humic substnces is hve prebiotic effect by incresing production of lctic responsible for some erpeutic rections occurring in cid, us incresing e prolifertion of beneficil tissues. The erpeutic properties of humtes hve been bcteri nd boosting e immune system [2]. Reserch described s ntibcteril, ntitoxic, ntiulcerogenic, hs indicted t supplementtion wi MOS results in ntirritic, ntillergic, immunomodultory nd ntin improvement in intestinl morphology nd intestinl inflmmtory [15]. Dietry humtes my lso influence enzyme ctivity [11]. These workers supplemented niml performnce indirectly by ltering e sorghum/lupin-bsed diet wi four levels (0, 1, 3 nd 5 gstrointestinl ecosystem since e microbil fctors t g/kg) of commercil MOS (Bio-Mos ) nd reported influence competition in e gstrointestinl trct centre beneficil effect on jejunl villi height nd ctivities of on e respective orgnisms present nd eir bility to mltse, leucine minopeptidse nd lkline scvenge nd grow on vilble nutrients [16]. Certin phosphtse. By blncing e intestinl environment modi?ed forms of humic cid hve been shown to possess nd stimulting e immune response, MOS hve ntivirl ctivities ginst herpes simplex nd humn been shown to increse grow in broilers [11]. Mnnn immunode?ciency viruses [17]. At concentrtion of 100 oligoscchrides lso reduces pogen coloniztion ìg/ml of mmonium humte in vitro hve been described by cting s n nlogous receptor to type I fimbrie nd resulting in e successful use of is gent s topicl by decresing e number of binding sites in e tretment for herpes virus-induced skin diseses [13]. intestine [12]. Inflmmtory immune responses re Schneider et l. (1996) [18] reported on e nti-hiv ssocited wi nutrient mobiliztion during growing ctivity of synetic humte nlogues derived from phse nd lower feed intke. Thus, dietry hydroquinone. These compounds inhibited HIV-1 immunomodultors t increse humorl immunity nd infection of MT-2 cells wi n impressively low IC 50 of decrese e immunologicl stress will ffect broiler ng/ml. The infectivity of HIV prticles ws performnce [13]. There is lso some evidence t MOS inhibited by interference wi e CD4-induced proteolytic my hve ntioxidnt properties, possibly leding to clevge of e V3-loop of virion gp120su. Humte improvement in ctivity of ntioxidnts such s should inhibit pogenic bcteril nd moulds grow, glutione peroxidse (GSH-Px) nd superoxide us decresing levels of mycotoxin, which should led to desmutse (SOD). It is importnt to tke ll of ese improved gut hel [19]. Humic cids hve lso been effects of MOS on bo microflor popultions nd e successfully used s nti-inflmmtory gents becuse of immune system when looking t its effects on disese eir locl nti-inflmmtory, hyperemic nd nlgesic sttus of birds. In ddition, previous reports suggest t properties [20]. MOS supplementtion resulted in significnt improvement In lterntive medicine, it is long time t HU ws in ntibody responses in lyers [2]. Defense cells in e known to improve e function of e immune system. We liver detect some molecules known s pogens erefore investigted e effects of dietry ssocited moleculr pttern t re specific of ech supplementtion of HU nd MOS preprtion on microorgnism. These molecules include yest cell wll, ntibody production ginst NDv s n indictor of peptoglycns, lipopolyscchrides nd glycolipids [4]. humorl immunity of birds. 780

3 MATERIALS AND METHODS by hemgglutintion-inhibition (HI) test. At e ge of 10 dys, ll chicks were vccinted wi Hitcher A totl of 384 one-dy-old broiler chicks (not sexed) B1 ND vccine by eye dropper nd bivlent killed were dpted for 3-d period before commencement of 1 vccine (ND plus AI) by inocultion ccording to e e tril. During is period, e birds were submitted to recommendtion of e mnufcturer. Blood smples conventionl broiler chicken mngement nd housed in were collected every week from e wing veins floor pens in n environmentlly controlled broiler house of individul chickens in ll groups nd eir ser wi litter floors. They received commercil broiler were seprted nd inctivted t 56 c for 30 min nd strter diet, (similr to e negtive control diet in e kept t-20 c until nlysis for e level of ND ntibody respective experiments, formulted to meet or exceed e [22]. nutritionl requirements of broilers, s recommended by The experiment lsted for 42 dys. Dt re expressed e Ntionl Reserch Council (NRC), 1994 [21]. Lighting s men ±SEM nd were evluted wi ANOVA for ws provided for 23 h/d. At 7-d of ge, 384 chicks of complete rndomized design, using e generl liner similr weights were rndomly ssigned to 32 pens in models procedure of SAS softwre [23]. The tretment e sme broiler house used for e dpttion period. mens wi significnt differences were compred by The birds were ssigned to e following tretment using Duncn's new multiple rnge tests. groups (in 3 replictes) of 12 chicks: 1) negtive control (CTL-), neier vccinted nor given MOS nd HU; 2) RESULTS AND DISCUSSION positive control (CTL+) or broilers were vccinted ginst infectious bursl disese virus, MOS nd HU-free Effect of dietry HU nd MOS on ntibody titers diet; 3, 4 nd 5) CTL+ plus diet supplemented wi 0.1, 0.2 ginst ND virus is shown in Tble 1. On e 7 nd 14 nd 0.3% HU (HU = Frmgültor DRY humte), dy of e study, ere ws no difference mong ntibody respectively; 6, 7 nd 8) CTL+ plus diet supplemented titers of experimentl groups. Animls of CTL-groups wi 0.1, 0.2 nd 0.3% MOS; respectively. Ech kilogrm showed reduction (P<0.01) in ntibody titers ginst ND of humte contined 160 mg polymeric polyhydroxy cid virus s compred to e CTL+ t 21, 35 nd 42 dys of (humic, fulvic, ulmic nd humtomelnic cids), mg ge. During dys 21 to 35 supplementl HU nd MOS SiO2 nd oer minerls (Mn, 50 mg; Zn, 60 mg; Fe, 60 mg; improved ntibody titers ginst ND virus. The results Cu, 5 mg; Co, 0.2 mg; I, 1 mg; Se, 0.5 mg nd Al, N, K, Mg presented in is study demonstrted t e HU dose nd P in trce mounts. dependently potentited e ntibody response ginst The bsl control diet ws formulted nd ND virus in broilers. compounded to meet e nutritionl requirements of Compred wi e positive control (CTL+), commercil broilers during e strter period (0 to 3 wk: supplementtion wi MOS t 0.2 % incresed ntibody corn grin, 61.5 kg; soyben mel, 30.5 kg; soyben oil, 3.1 titers, its effect t oer inclusion levels (0.1 % or 0.3 %) kg; oyster shell, 1.2 kg; diclcium phosphte, 1.95 kg; ws lower, us t dy 21 nd 35 of ge supplementtion minerl mix, 0.25 kg; slt, 0.48 kg; vitmin mix, 0.25 kg; L- wi MOS t 0.2 % showed best effect on ntibody titers Lysine, 0.2 kg; DL-Meionine, 0.27 kg to mke up 21.9% ginst ND virus. At dy 35 ntibody titers ginst ND crude protein; 3100 kcl/kg ME; 1.2% Lysine; 0.94% virus in CTL+ group t hs ny dditive, differ Meionine; 0.9% Clcium; nd 0.45% Avilble significntly wi oer groups. phosphorus) nd grower period (4 to 6 wk: corn grin, 65.2 No significnt differences on ntibody titers were kg; soyben mel, 27.4 kg; soyben oil, 3.5 kg; minerl mix, found between e HU-treted birds nd e HU-treted 0.25 kg; vitmin mix, 0.25 kg; slt, 0.4 kg; oyster shell, 1.1 birds in e entire experimentl period, but levels of kg; diclcium phosphte, 1.6 kg; DL-Meionine, 0.2 kg to dditives hve vrious effect. Present study s well s mke up 19.58% crude protein nd 3180 kcl/kg ME; previous studies shows t HU nd MOS hve revivl 1.13% Lysine; 0.75% Meionine; 0.85% Clcium nd effect on immune responses. The feeding of mnnn 0.42% Avilble phosphorus). oligoscchrides nd humte t level of 0.2 nd 0.3% Prior to division of chicks (1-d-old) in to vrious respectively in e rtion incresed e ntibody groups, blood smple ws tken from ech bird nd e production ginst ND virus in broilers from 21 to 35 dys titers of mternl ntibody ginst Ndv were mesured of ge. 1 Newpsol 102, Inctivted W/O Emulsion ND+AI Vccine, Psouk Biologicl Co. 781

4 Tble 1: Effect of mnnn oligoscchrides nd humte on NDV ntibody titers in broiler chicks from 7 to 35 dys of ge tretment ND ntibody titer Vc HU % MOS % 7 dy 14 dy 21 dy 35 dy 42 dy c 0.77 d 0.00 b ** b 2.33 c b 3.33 b b 4.23 b b 4.11 b b 3.67 b b 4.33 b VAC: vccintion; HU: humte; MOS: mnnn oligoscchrides., b, c Mens in row wi no common superscripts differ significntly (P = 0.01). Ech vlue represents e Mens of e 12 chicks per tretment. Vlues represent e men wi SE in preneses for ech tretment. ** The negtive control groups did not contin detectble levels of ntibody titers. Vertebrtes re cpble of two types of immune These results demonstrted e positive influence of HU responses, innte nd dptive. The dptive (lso clled nd MOS on e response to vccintion of e chicken s e specific or cquired) immune response involves immune system. This effect on ntibody titers might hve recognition nd effector ctions t re highly specific to been due to influence of e MOS on immune system; e precise foreign entity t hs triggered e response. improved intestinl bsorption of some nutrients, such s The innte nd dptive immune responses do not Zn, Cu, Se; or bo effects. Improved intestinl lumen operte in isoltion nd ech is dependent on or hel ws observed in MOS fed mle pullets [25] s well enhnced by elements of e oer. Immuniztion in e s in 1-wk old mle broilers fed Scchromyces cerevisie clinicl sense is e process of mnufcturing immune (SC) cell wll [9]. Ingestion of diets contining MOS defense, which is, rtificilly helping e body to defend resulted in higher titers, in comprison to birds which it using e effectors nd mechnisms of e immune were fed diets wiout MOS, specificlly during e weeks system. In ctive immuniztion, which we commonly cll in which titers tended to decrese. These findings re in vccintion, modified form of e nturl immunogen greement wi ose reported by Shshidhr nd (e vccine) is dministered so t e host develops Devegowd (2003) [4]. In greement wi is study, specific B nd T memory cells t cn ct ginst e beneficil effects of yest cell wll (YCW) on humorl nturl immunogen. Vccintion is n effective mens to immunity were lso observed in broiler chicks [7]. Much prevent nd/or reduce e dverse effects of specific of e nture of mechnism ccountble for diseses in poultry. Control of virl diseses is dependent immunomodultion ssocited wi e MOS remins to upon prevention rough snittion nd biosecurity nd be delineted. One hypoesis is t defense cells in e by vccintion. The immune response fter vccintion is gut-ssocited lymphoid tissue (GALT) detect e n elegnt tool for studying e effect of diet dditive presence of microbes by recognizing molecules unique to bo in humn nd niml subjects. Antibody titer microorgnisms t re not ssocited wi host cells. responses hve been used s mesures of humorl These unique molecules re clled Pogen Associted immune sttus of birds [24]. In our study, for evluting Moleculr Ptterns (PAMP). These include yest cell e effect of MOS nd HU on immune system wll components such s mnnn nd glucn long experimentl, chicks were vccinted wi Hitcher B1 ND wi oer microbil molecules such s peptidoglycn, vccine by eye dropper nd bivlent killed vccine (ND lipopolyscchride nd glycolipids [3]. Mnnn nd plus AI) by inocultion on dy 10. glucn of e yest cell wll my bind to pttern- Antibody titer ws significntly incresed in e HU recognition receptors on vriety of defense cells of e nd MOS group in is tril (Tble 1) nd suggests t GALT nd in turn ctivte immune defenses such s HU nd MOS my lso be influencing systemic immunity. phgocytosis, e lterntive complement pwy nd 782

5 e lectin pwy. In poultry, reports pertining to e contining HU resulted in more high titers, in effect of MOS inclusion in e breeder diet on mternl comprison to birds which were fed diets wiout ntibody titers in progeny re sprse. Yest cell contins dditives (MOS or HU) specificlly during e weeks in 1,400 units/kg of phytse nd Liu et l. (2008) [26] which titers tended to decrese. Present study s well s reported t ppliction of phytse in nutritionlly previous studies show t HA hve revivl effect on mrginl diets could enhnce lymphocyte numbers nd immune responses. Dekker nd Melden (2003) [15] e serl nd mucosl ntibodies of broilers, suggesting reported t Oxihumte (99% purified HA) incresed e t phytse my hve role in gstrointestinl hel prolifertive response of phytohemgglutinin (PHA) nd immune competence. Severl workers [1] reported t stimulted lymphocytes in dose-relted mnner. Also, SC improved e efficcy of e immune system, improved oxihumte incresed e expression of CD25 significntly intestinl lumen hel nd incresed digestion nd on PHA-stimulted cells. Oxihumte cused sttisticlly bsorption of nutrients, which resulted in better significnt increse in IL-2 nd IL-4 production by performnce. Supplementl yest cell (YC) incresed P stimulted lymphocytes. A slight but not significnt nd C digestibility. Erlier study [25] lso reported increse in IL-6 production by oxihurnte ws lso improved utiliztion of phosphorus or clcium in observed. An increse in IL-2 is necessry for cellphosphorus-insufficient or sufficient diets of poultry medited (TH l-type) immunity, s well s n increse in IL-4 when YC ws supplemented. nd IL-6, is necessry for humorl (TH 2-type) immunity, Mnnn oligoscchrides lso my modulte humorl indicte t is compound cn be n effective immunity ginst e coloniztion of specific pogens, immunostimulnt which my prove suitble for ppliction llowing em to be presented to immune cells s by ptients suffering from virl nd bcteril infections. ttenuted ntigens due e production of short chin Dekker nd Melden (2003) [15] reported t chickens ftty cid (SCFA) derived from MOS fermenttion t treted wi 100 mg oxihurntelkg/dy showed two to decreses intestinl ph nd reduces e presence of four fold increse in titers ginst Newcstle disese. This pogenic bcteri. Butyrte is one of e SCFA result indictes e bility of ese compounds to produced nd its deficiency is ssocited wi intestinl stimulte e humorl immune response. Vn Rensburg et trophy nd functionl fils, including decresed l. (2002) [17] found t is product stimultes immune response, however, n incresed butyrte supply lymphocyte prolifertion, importnt for cell medited leds to e development of intestinl epielium, cell immunity necessry to cope wi opportunistic infections, differentition nd incresed immune response, probbly by incresing e production of e Interleukin-2 (IL). due to e higher number of T-cells in e intestine [4]. It Joone et l. (2003) [20] found t oxihumte incresed e ws proposed t oligoscchrides in e MOS could in vitro prolifertive response of PHA-stimulted humn bind to viruses nd work s djuvnts of vccines to lymphocytes, from concentrtion of 20 µg oxihumte/ml increse e titers of ntibody in MOS-treted birds [3]. nd upwrds. This implies t MOS my stimulte e humorl immune The effects of HU on ntibody titers re due to its system to produce more ntibodies. Spring et l. (2000) [1] ntivirl properties, phgocytic ctivity of leukocytes, reported t pogens wi e mnnose-specific type-1 dhesibility nd ctivtion of neutrophils [27], improving fimbrie, such s E. coli nd Slmonell, re ttrcted to nutritive vlue of feed [16], bility to block coloniztion of e mnnns nd redily bound wi e crbohydrte pogens in e gstrointestinl trct [13] nd improving insted of ttching to intestinl epielil cells, Mnnn immune functions [27]. The immune response is medited oligoscchrides lso decresed e coloniztion by by mcrophges ctivtion since mcrophges hve been Escherichi coli nd incresed Lctobcillus in e ctivted by HU, s demonstrted by Chen et l. (2003) chicken gut. Antibody titers vried wi e levels of [27]. An ntivirl property, t concentrtion of 100 ìg/ml MOS supplemented nd ws best when 2g/kg of MOS of mmonium humte (e mmonium slt of humic cid) ws supplemented under e experimentl conditions of in vitro hs been described by Thiel et l. (1981) [28]. On is study lough is difference ws not significnt. In e oer hnd, e ntimicrobil effects of HU hve been summry, e results of e current study indicte t described in e literture [29]. Visser (1985) [14] MOS improves immune functions of broilers. suggested t humic cids my induce chnge in In is study e inclusion of HU resulted in metbolism, llowing microorgnisms to utilize different incresed ntibody titers ginst NDV in e 21 (P<0.01) substrtes which could ccount for our observed shift in nd 35 (P<0.01) dys of ge. Ingestion of diets E coli popultions. 783

6 R et l. (2006) [30] hve shown t e reltive 8. Jensen, G.S., K.M. Ptterson nd I. Yoon, Yest weights of e burs of fbricius incresed in culture hs nti-inflmmtory effects nd specificlly birds given 0.25% HU suggesting possible ctivtes NK cells. Comp. Immunol. Microbiol. Infect. immunostimultory effect t hs been considered Dis., 31: to be n effect of HU. Humte dministrtion for 9. Sntin, E., A. Miork, M. Mcri, M. Grecco, 8 weeks improved grow performnce in e newborn J.C. Sncheez, T.M. Okd nd A.M. Mysk, kids nd lso hd n effect t 21 d on skin rection to Performnce nd intestinl mucos development of phytohemgglutinin, possibly n effect on cell-medited broiler chickens fed diets contining Scchromyces immune response [31]. cerevisie cell wll. J. Appl. Poult. Res., 10: In conclusion ccording to founding of present 10. Zhng, A.W., B.D. Lee, S.K. Lee, K.W. Lee, G.H. An, study, HU nd MOS cn modulte e ntibody response K.B. Song nd C.H. Lee, Effects of yest ginst Newcstle disese vccine in broiler; however is (Scchromyces cerevisie) cell components on wrrnts furer evlution of humic substnces nd grow performnce, met qulity nd ilel mucos mnnn oligoscchrides on oer vccines in different development of broiler chicks. Poultry Science, species nd evlution of moleculr spects. 84: Iji, P.A., A.A. Ski nd D.R. Tivey, Intestinl REFERENCES structure nd function of broiler chickens on diets supplemented wi mnnn oligoscchride. J. Sci. 1. Spring, P., C. Wenk, K.A. Dwson nd K.E. Newmn, Food Agric., 81: The effects of dietry mnnn oligoscchrides 12. Oliveir, M.C., D.F. Figueiredo-Lim, D.E. Fri Filho, on cecl prmeters nd e concentrtions of enteric R.H. Mrques nd V.M.B. Mores, Effect of bcteri in e cec of Slmonell-chllenged broiler mnnn oligoscchrides nd/or enzymes on chicks. Poultry Science, 79: ntibody titers ginst infectious bursl nd 2. Cotter, P.F., A. Mlzone, B. Pluch, M.S. Lilburn nd Newcstle disese viruses. Arq. Brs. Med. Vet. A.E. Sefton, Modultion of humorl immunity Zootec., 61(1): in commercil lying hens by dietry prebiotic. 13. Klocking, R., Humic substnces s potentil Poultry Science, 79(Suppl.1): 38 (Abstr.). erpeutics. In: Humic Substnces in e Globl 3. Bllou, C.E., A study of e immunochemistry Environment nd Implictions on Humn Hel, of ree yest mnnns. J. Biol. Chem., Eds., N. Senesi nd T.M. Mino. Elsevier Science, 245: pp: Shshidhr, R.G. nd G. Devegowd, Effect of 14. Visser, S.A., Effect of humic cids on numbers dietry mnnn oligoscchride on broiler breeder nd ctivities of microorgnism wiin physiologicl production trits nd immunity. Poultry Science, groups. Org. Geochem., 8: : Dekker, J. nd C.E. Medlen, Oxihumic cid nd 5. Sonmez, G. nd M. Eren, Effects of its use in e tretment of vrious conditions. United supplementtion of zinc bcitrcin, mnnn Sttes Ptent: oligoscchride nd probiotic into e broiler feed on 16. Kocbgli, N., M. Alp, N. Acr nd R. Khrmn, morphology of e smll intestine. Vet. Fk. Derg The effects of dietry humte supplementtion Uludg Univ., 18: on broiler grow nd crcss yield. Poultry Science, 6. Newbold, C.J., R.J. Wllce, X.B. Chen nd 81: F.M. Mcintosh, Different strins of 17. vn Rensburg, C.E.J., J. Dekker, R. Weis, T.L. Smi, Scchromyces cerevisie differ in eir effects on vn E.J. Rensburg nd J. Schneider, ruminl bcteril numbers in vitro nd in sheep. J. Investigtion of e nti-hiv properties of oxihumte. Anim. Sci., 73: Chemoerpy, 48: Go J., H.J. Zhng, S.H. Yu, S.G. Wu, I. Yoon, J. 18. Schneider, J., R. Weis, C. Mnner, B. Kry, Quigley, Y.P. Go nd G.H. Qi, Effects of A. Werner, B.J. Seubert nd U.N. Riede, yest culture in broiler diets on performnce Inhibition of HIV-1 in cell culture by synetic nd immunomodultory functions. Poultry Science, humte nlogues derived from hydroquinone: 87: mechnism of inhibition. Virology, 218:

7 19. Ghhri, H., R. Hbibin nd M. Abdolhfm, 26. Liu, N., Y.J. Ru, A.J. Cowieson, F.D. Li nd Evlution of e efficcy of esterified X.C.H. Cheng, Effects of Phytte nd Phytse glucomnnn, sodium bentonite nd humic cid on e performnce nd immune function of broilers to meliorte e toxic effects of fltoxin in broilers. fed nutritionlly mrginl diets. Poultry Science, Turkish Journl of Veterinry Animl Science, 87: (4): Chen, C.H., J.J. Liu, F.J. Lu, M.L. Yng, Y. Lee nd 20. Joone, G.K., J. Dekker nd C.E.J. Vn Rensburg, T.S. Hung, The effect of humic cid on e Investigtion of e immunostimultory dhesibility of neutrophils. Thromb. Res., 108: properties of oxihumte. Verlg. Z. Nturforsch., 28. Thiel, K.D., B. Helbig, R. Klöcking, P. Wurtzer, 58: M. Sprössig nd H. Schweizer, Comprison of 21. Ntionl Reserch Council (NRC), Nutrient e in vitro ctivities of mmonium humte nd Requirements of Poultry. 9 rev edn. Ntionl of enzymti clly oxidized chlorogenic nd cffeic Acdemy Press. Wshington, DC. cids ginst type 1 nd type 2 humn herpes virus. 22. Alln, W.H. nd R.E. Gough, A stndrd HI test Phrmzie, 36: for Newcstle disese vccintion nd chllenge. 29. EMEA, Humic cids nd eir sodium slts, VET. REC., 95: summry report. Committee for Veterinry 23. SAS Institute, SAS User s Guide. Version 8.02 Medicinl Products. Eur. Agency Evl. Med. Prod. ed. SAS Institute Inc., Cry, NC Skln, D., D. Melmed nd A. Friedmn, Accessed June 10, The effect of vrying levels of dietry vitmin A on 30. R, N.C., W.E. Huff nd G.R. Huff, Effects of immune response in e chick. Poultry Science, humic cid on broiler chickens. Poultry Science, 73: : Brdley, G.L., T.F. Svge nd K.I. Timm, Agzzi, A., G. Ciglino, G. Mncin, G. Svoini The effects of supplementing diets wi nd V. Dell Orto, Effects of dietry humtes on Scchromyces cerevisie vr. boulrdii on mle grow nd n spect of cell-medited immune poult performnce nd ilel morphology. Poultry response in newborn kids. Smll Ruminnt Reserch, Science, 73: :