Residual Feed Intake and its Effect on Cell-Mediated Immunity in Laying Hens Given Different Propolis Levels

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1 Interntionl Journl of Poultry Siene 7 (11): , 008 ISSN Asin Network for Sientifi Informtion, 008 Residul Feed Intke nd its Effet on Cell-Medited Immunity in Lying Hens Given Different Propolis Levels A. Gll, A.M.H. Ahmed, W.A.H. Ali, M.H. El-Snhoury nd Hedi E. Ahmed 1 Deprtment of Poultry Prodution, Fulty of Agriulture, Ain Shms University, Ciro, Egypt Deprtment of Turkey nd Wterfowl, Animl Prodution Reserh Institute, Centrl Agriulturl Reserh, Rits, Egypt 3 Animl Prodution Reserh Institute, A.R.C., Ministry of Agriulture, Egypt Astrt: The min im of this study ws to investigte the reltionship etween residul feed intke nd some hemtologil prmeters in lying hens fed different level of Propolis. Residul feed intke is used s trit to distinguish effiient nd non-effiient lying hiken. A Phytohemgglutinin-P (PHA-P) ssy nd white lood ells differentition were used to evluting the immune response of the lying hens fed different propolis levels. The present result indited tht the lying hens fed different levels of Propolis hd signifintly hyper responder to PHA-P injetion, higher lymphoytes ount nd lower heterophils ount when ompred with ontrol-group. With respet to residul feed intke, it ws generlly notied tht the oserved feed onsumption vlues were somewht losely to expet feed onsumption vlues for hens fed diet ontining 100 or 150 mg Propolis ompred to remining tretments. Also, the results reveled tht the equtions lulted for lying hens fed diet dding 100 or 150 mg Propolis hd etter rte of determintion (R ) ompred to other fed 0 or 50 mg Propolis. In ordne to orreltion oeffiients, it ould e notied tht the wttle swelling mesured t 4, 48 nd 7 h post PHA-P injetion were signifintly positive orrelted with residul feed onsumption in supplementl propolis t 100 mg. Similr trend ws oserved in 150 mg supplementl propolis lying t 7 h post PHA-P injetion. With respet to white lood ells differentition, the present results showed tht signifintly negtive reltionship etween RFC nd heterophils perentge in ll tretment groups. Inversely, the reltionship etween RFC nd lymphoytes perentge ws signifint nd positive in ll tretment groups. Finlly, it ould e onluded tht the Propolis supplementtion improve the immune sttus of lying hens vi minimizing the residul feed intke. Key words: Residul feed intke, immunologil prmeters, lying hens INTRODUCTION Lying hens n e onsidered s eing genetilly progrmmed for effiient Egg Mss (EM) prodution. There is, however, lot of vrition in effiieny etween individuls within popultion, s shown y vrition in feed onsumption tht nnot e explined y metoli Body Weight (BW), ody weight gin nd Egg Mss (EM). This vrition in feed onsumption is refleted in differenes in Residul Feed Intke (RFI) (Luiting nd Urff, 1991). RFI is defined s the differene etween oserved Feed Intke (FI) nd FI predited from metoli BW (representing mintenne) nd BW gin nd EM (oth representing prodution). RFI is, thus, mesure for feed effiieny: hikens with low RFI (R-) need less feed to reh the sme BW nd prodution level nd re, therefore, more effiient produers thn hikens with high RFI (R+). The phenotype of n niml is funtion of its genotype nd present nd pst environments. Environmentl ftors suh s nutrition, feeding mngement nd temperture my influene growth nd modify sensitivity to infetious gents nd immunoresponsiveness (Zulkifli et l., 1994). Immunoompetene nd growth re influened y geneti nd non-geneti ftors, only reently hs evidene ppered showing negtive reltionship etween ody weight nd immunoompetene in Leghorns (Mrtin et l., 1990), roilers (Qureshi nd Hvenstein, 1994) nd rown egg lyers (Kreukniet et l., 1994). In roiler reeding there is further omplition euse lthough roilers re rered under d liitum feeding, the mngement of roiler reeders involves restrition in feed intke to ontrol over-onsumption nd its deleterious effets on helth nd reprodution (Ktnf et l., 1989). As result of this husndry, reserh results hve umulted on effets of vrious feeding regimens on ody weight (Roinson et l., 1993) nd feed intke nd diet seletion (Fores, 1995). Immune stimultion (Klsing et l., 1987) nd injetion of hikens with IL-1- ontining superntnt from teril endotoxin stimulted hiken HD11 mrophges (Klsing et l., 1987) indued redution of growth nd feed utiliztion in hikens nd elerted musle protein degrdtion. In this study, we evlute the reltionship etween residul feed intke nd ell medited immunity of lying Hy-line hens fed different levels of Propolis. 1105

2 Int. J. Poult. Si., 7 (11): , 008 MATERIALS AND METHODS This experiment ws rried out t poultry reeding frm, Poultry Prodution Deprtment, Fulty of Agriulture, Ain Shms University. One hundred nd twenty 46 -week-old, Hy-line White strin were rndomly ssigned to 4 groups of 30 hens eh. The hens were individully housed in individully ges, on 16 h light shedule. Chikens in ll groups were rered under the sme environmentl, mngeril nd hygieni onditions. Feed nd wter were supplied d liitum. The hens reeived typil lyer diet ontining 800 ME kl/kg nd 18% CP to meet or slightly exeed the nutrient requirement reommended y NRC (1994). The hens were fed sl diet (ontrol) or sl diet ontining 50, 100 nd 150 mg Propolis/kg. Mesurements nd oservtions: Body weight ws individully reorded t 46, 50 nd 54 weeks of ge. Also, numer nd weight of eggs were reorded dily from the weeks of ge. Internl nd eggshell qulity ws determined t 54 weeks of ge. The egg length (long xis) nd width (short xis) were mesured with the eletroni liper. The width to length rtio ws shown in perentge points nd onstituted the egg shpe index. The height of thik lumen (H) nd the egg Weight (W) were used to lulte Hugh units from the formul of Willims (1997): 0.37 HU = 100 log (H W ) where: H = Thik lumen height. W = Egg weight. Yolk dimeter long the hlze line ws determined with the liper (mm). The eggshell, fter the removl of the egg ontent, ws dried. Susequently the eggshell ws weighed to the nerest 0.01 g. Eggshell thikness without inner memrnes ws mesured (mm) with the mirometer. The lumen weight ws lulted from the differene etween the entire egg weight nd the yolk nd eggshell weight. The ontents of yolk, lumen nd the eggshell were expressed s perentges from the weight of fresh egg. The reking strength ws mesured ording to Fthi nd El-Shr (1996) whih ssessed the resistne of the egg to rushing. At 54 weeks of ge, lood smples were tken from the rhil vein into heprinized tues for ll irds. Plsm ws otined from the lood smples y entrifugtion for 10 min t 4000 rpm nd ws stored t -0 C until the time of nlysis. The frozen plsm ws llowed to thw t room temperture prior to nlysis. Plsm totl protein, lumin nd holesterol were determined y enzymti olorimetri methods using ville ommeril kits. The plsm gloulin ws lulted s the differene etween plsm totl protein nd lumin. A Phytohemgglutinin-P (PHA-P) injetion ssy (Cheng nd Lmont, 1988) ws used to evlute in vivo T-ellmedited immune response of Hy-Line lying hens. Birds were injeted intrdermlly in the wttle with 0.5 mg of PHA-P (Sigm Chemil Co., St. Louis, Missouri) in 0.1 ml of Phosphte Buffered Sline (PBS) fter mrking the injetion site. The thikness of wttle ws mesured (to nerest 0.01 mm]) t 0, 4, 48 nd 7 h fter PHA-P injetion. Wttle swelling ws lulted s the differene etween the thikness of the wttle prior to nd fter injetion of PHA-P. At 54 week of ge, lood smples were otined from eh tretment for Heterophil (H) nd Lymphoyte (L) enumertion sed on the proedures of Gross nd Siegel (1983). Briefly, one drop of lood eing smered on eh of glss slides. The smers were stined using Wright s stin. Two hundred leukoytes, inluding grnulr (heterophils) nd nongrnulr (lymphoytes) ones, were ounted on different mirosopi fields representing 00 ells nd the heterophil to lymphoyte rtio ws lulted. Computing dt nd sttistil nlysis: The feed onsumption for hens ws predited to derive regression eqution ording to tretment. Residul Feed Consumption (RFC) ws lulted s the Differene Between Oserved (OFC) nd Expeted Feed Consumption (EFC) for eh experimentl hen. Eh tretment hd its own regression oeffiients ording to the following eqution: where: EFC Bw i EFC = BW i + EM i + )W i + d = Expeted feed onsumption of hen I grms). = Men metoli ody weight of hen I (kg ). = Egg mss prodution of hen I (grms). Em i )w i = Body weight gin., nd = Prtil regression oeffiients. d = Interept. All lultions nd nlyses were mde using Generl Liner Models (GLM) proedure of SAS User's Guide, 001. Correltion oeffiients of RFC with produtive trits were estimted for eh strin using the PROC CORR proedure of SAS. RESULTS AND DISCUSSION Cell-medited immunity, some hemtologil prmeters nd produtive trits of lying hens fed different levels of Propolis re summrized in Tle 1. The PHA intrdermlly retion, T-lymphoytedependent response, hs een well reserhed nd hs een shown to e relile inditor of in viv 1106

3 Int. J. Poult. Si., 7 (11): , 008 Tle 1: Cell-medited immunity, some hemtologil prmeters nd produtive trits of lying hen fed different levels of Propolis Propolis level (mg/kg) Pooled S.E.M Pro. Cell medited-immunity 4 h post PHA-P injetion d h post PHA-P injetion d h post PHA-P injetion d White lood ells Heterophils Lymphoytes Hemtologil prmeters Totl plsm protein (g/dl) Gloulin (g/dl) Cholesterol (mg/dl) Produtive prmeters Oserved feed onsumption Feed onversion rtio Egg numer Egg weight Alumen (%) ns Yolk (%) ns Shell (%) Shell thikness Breking strength d Mens with different letters within row re signifintly differed ellulr immunity in poultry (Goto et l. 1978). The skin infeted with virulent NDV nd susequently treted response reflets omplex series of physiologil either with propolis or honey if ompred with the events suh s mitogen-reeptor nd lymphoyte- infeted groups only. It ws ler tht, propolis ts mrophge intertions, relese of hemil tively s ntivirl gent thn honey. The tretment with meditors, ellulr prolifertion nd hnges in propolis nd honey of NDV infeted hiken groups vsulrity (Chndr nd Newerne, 1977). indued inrese in the ntiody titres nd phgoyti Histologilly, PHA is strongly mitogeni to T- perentge. The inoultion of different ntigens in the lymphoytes nd intrdermli injetions eliit footpd of sensitized nd non-sensitized hikens mrophge infiltrtion nd dense perivsulr indued different degrees of footpd thikness s well umultions of lymphoytes 4 h post-injetion in s ellulr nd vsulr retion depending on the type hikens (Goto et l., 1978; MCorkle et l., 1980). The of inoultion with NDV ntigen. In ordne to white inresed infiltrtion y sophils nd eosinophils 4 h lood ells differentil, it ould e notied tht the post-injetion hs een desried s utneous Propolis levels t 100 nd 150 mg signifintly sophil hypersensitivity response (Stdekerm et l., deresed heterophils ount nd signifintly inresed 1977). The present result showed tht the speifi lymphoytes ount ompred to ontrol-fed group. In responses to PHA-P injetion t ll times showed irds, the heterophil re phgoyti ells whose min is mrked differenes etween tretment groups, with protetion ginst invding miroorgnisms, wheres supplementl Propolis lying hens t ll levels rehing primry funtions of lympho-involve ell-medited nd muh higher responder thn ontrol group. Propolis humorl immunity. Heterophils inrese nd ording to reserh hs shown to e effetive ginst lymphoytes derese when re stressed, so tht the vriety of teri, viruses, fungi nd molds. It hs rtio etween them is index of response to stressor een shown to e non-speifi immunostimulnt. The (Gross nd Siegel, 1985). In ordne to H/L rtio, our delyed hypersensitivity skin test using propolis s results showed tht the propolis supplementtion t sensitizing ntigen showed speifi stimultion to nd 3g/kg diet signifintly inresed the H/L rtio of propolis fter 7 h fter inoultion with speifi ntigen. lying hens. The H/L rtio is reognized mesure of Egyptin propolis gve the typil delyed stress in irds (Mxwell, 1993) tht hs eome hypersensitivity when inoulted to the sensitized vlule tool in stress reserh espeilly when hikens. The thikness index ws 0.90 mm thikness omined with the onveniene nd repetility of if ompred with non-sensitized ontrol group 0.1 mm utomted lood ell ounts. thikness (Hegzi et l., 1996). Hegzi et l. (1995) With respet to hemtologil prmeters, it ould e studied the effet of some ee produts on immune speulted tht the lying hens fed high Propolis levels response of hiken infeted with virulent NDV. They (100 nd 150 mg) were signifintly inresed totl found tht, the mortlity rte ws redued in-groups plsm protein nd plsm gloulin ompred to 1107

4 Int. J. Poult. Si., 7 (11): , 008 ontrol-fed group. Inversely, the Propolis supplementl t high levels signifintly deresed plsm holesterol. The low level of Propolis ws intermedited in ll ses. Similr reports were drwn y Giurge et l. (1981). They indited tht dily dministrtion of propolis extrt to hikens hnged the lood onentrtion of holesterol, totl proteins nd mino id. Also, Propolis stimulted mmmlin tissue regenertion, s it used strong tivtion of mitosis of ells ultured in vitro nd it enhned protein iosynthesis (Grys et l., 1986). The lying hens fed either 100 or 150 mg Propolis onsumed more feed ompred to other fed ontrol-fed group. However, the 50 mg Propolis level ws intermedited. Conerning feed onversion rtio, the present result indited tht the Propolis supplementtion t ll levels hd signifintly improved feed onversion rtio ompred to ontrol-fed group. The egg numer ws signifintly ffeted y Propolis supplementtion. Moreover, the egg numer ws grdully inresed with Propolis level inresing. The lying hens fed diet ontining 100 nd 150 mg Propolis signifintly inresed egg weight ompred to ontrol fed-group. The inrese feed intke nd egg mss in Propolis groups, resulting in signifintly improve feed onversion rtio ompred to ontrol-group. Shlmny nd Shivzd (006) reported tht hiks fed Propolis ontining diets onsumed signifintly higher feed. Bonomi et l. (1976) found n inrese in feed intke when lying hens were fed Propolis versus ontrol groups. Also, they onluded tht inrese in feed intke in the Propolis groups my e due to improved irds helth nd higher pltility of Propolis diets due to mixture of resin, wx, honey nd vnillin ontent of Propolis. Ghislerti (1979) report dditionl weight gins for roiler hikens of up to 0% when 500 ppm of propolis ws dded to their diets. They sid tht this improved effet is prtilly due to its high ontent of flvonoids nd inrese feed intke of Propolis diets thn the ontrol. Experimentl work of Buhtel et l. (1983) showed tht Propolis supplementtion to the rtion of pullets improved feed onversion. This effet is due to high ontent of flvonoids nd helthy onditions of irds fed Propolis. Conerning internl nd eggshell qulity, it ould e notied tht there ws no signifint differene mong tretment groups for oth lumen nd yolk perentge. However, the eggs produed from lying hens fed diet ontining either 100 or 150 mg Propolis were signifintly higher eggshell thikness ompred to other groups. Also, the supplementl Propolis t high level (100 or 150 mg) signifintly inresed eggshell reking strength ompred to remining groups. Multiple regression eqution of feed onsumption: Oserved feed onsumption, egg mss, hnge in ody Tle : Coeffiients of prtil regression nd onstnt for lying hens fed different Propolis levels Propolis (mg/kg) Constnt EM )W BW R EM = egg mss, )W = ody weight hnge, BW = metoli ody weight, R = rte of determintion weight nd metoli ody weight for eh hen within eh tretment ws used to estimte the regression oeffiients re listed in Tle. The results reveled tht the eqution lulted for lying hens fed diet dding 100 or 150 mg Propolis hd etter rte of determintion (R ) ompred to other fed 0 or 50 mg Propolis. Tht is men, the figures of RFC lulted from these equtions re more relile nd hve highly pplile prospetive. The present results reveled tht the regression of ody weight on feed intke my differ etween tretment groups, ut the ury of feed intke preditions is independent of the power used to express metoli ody weight. Pirhner (1985) reported tht differenes in Oserved Feed Consumption (OFC) nd Expeted Feed Consumption (EFC) re used y vriility in severl ftors, suh s omposition of produt (eggs), ody weight hnge, food spillge, metoli rte nd in the ility to synthesis egg nd ody onstituents. Figure 1 illustrtes the oserved ginst expeted feed onsumption for eh hen. While, Fig. depits the residul feed onsumption vlue for eh hen. It ws generlly notied tht the oserved vlues were somewht losely to expeted vlues for hens fed diet ontining 100 or 150 mg Propolis ompred to remining tretments. This djeny ws refleted on RFC, where it ws more onsistent to zero line. The negtive vlues of RFC re desirle rther thn positive ones. The effiient hens whih hve negtive RFC figures were more frequent thn ineffiient ones. The results of El-Syed nd El- Hkim (1994) nd Hussein et l. (000) onfirmed tht the effiient irds were less tives, less het prodution, spent more time resting nd less time stnding thn ineffiient ird. The lst result explined y vn Eerden et l. (004). They reported tht seletion for higher effiieny of food use in lying hens might result in irds tht one less frustrted prior to lying t less stressfully. If redution in feed intke is n inditor of stress, then the oservtion tht feed intke of the R+ hikens deresed onsiderly fter trnsporttion, wheres feed intke of R-hikens remined lmost unffeted. Correltion oeffiient etween RFC, ell-medited immunity nd some produtive trits: Clultion RFC y phenotypi multiple regression nlysis is n 1108

5 Int. J. Poult. Si., 7 (11): , 008 Fig. 1: Oserved vs. expeted feed onsumption of lying hens fed different levels of propolis eptle lterntive in reeding progrm of no supplementl Propolis t 100 mg. Similr trend ws relile estimtes of geneti orreltion re oserved in 150 mg supplementl Propolis lying t 7 ville (Luiting nd Urff, 1991). This suggestion ws h post PHA-P injetion. With respet to white lood ells reported euse the uthor found the estimtes of differentition, it ould e notied tht signifintly geneti orreltion of RFC with the eonomi trits did negtive reltionship etween RFC nd heterophils not lerly differ from zero. Also, Tixier-Boihrd et l. perentge in ll tretment groups. Inversely, the (1995) found tht the geneti orreltion etween RFC reltionship etween RFC nd lymphoytes perentge nd the independent vriles used in the predition ws signifint nd positive in ll tretment groups. Vn eqution (metoli ody weight, hnge in ody weight Eerden et l. (004) onluded tht popultion of nd egg mss) were generlly low, whih onfirms the hikens from ommeril reed shows onsiderle vlidity of the seletion on phenotypi ssessment of vrition in RFI. Speifi ntiody prodution ginst RFC. Dt presented in Tle 3 showed tht the KLH, M. utyrium nd S. enteritidis lipopolyshride, orreltion oeffiients for some quntittive trits with however, is not influened y effiieny in terms of RFI. Residul Feed Consumption (RFC) nd ell medited R+ nimls my hve higher level of non-ntigen immunity. The wttle swelling mesured t ll times speifi ntiodies, s indited y the higher ntiody were negtively orrelted, with sttistilly signifint t response to Slmonell protein. 48 h post PHA-P injetion, with RFC in ontrol-group. Heterophils ount ws signifintly negtive orrelted Similr, trend ws oserved in 50 mg supplementl with residul feed intke in ll tretment groups. Propolis. The wttle swelling mesured t 4, 48 nd 7 Conversely, there ws signifintly positive reltionship h post PHA-P injetion were signifintly positive etween lymphoytes ount nd residul feed orrelted with residul feed onsumption in onsumption in lying hens fed diet ontining 100 or 1109

6 Int. J. Poult. Si., 7 (11): , 008 Fig. : Residul feed onsumption of lying hens fed different levels of propolis Tle 3: Phenotypi orreltion oeffiients mong residul feed onsumption, ell-medited immunity nd some produtive prmeters Propolis (mg/kg) Cell medited-immunity 4 h post PHA-P injetion * h post PHA-P injetion -0.58* * h post PHA-P injetion * 0.53* White lood ells Heterophils -0.51* -0.66** -0.70** -0.68** Lymphoytes ** 0.70** Hemtologil prmeters Totl plsm protein Gloulin * Cholesterol Produtive prmeters Oserved feed onsumption 0.81*** 0.9*** 0.76** 0.63** Feed onversion rtio 0.58* 0.93*** 0.6** 0.79** Egg numer * Egg weight Alumen (%) Yolk (%) 0.53* 0.60** 0.68** 0.59** Shell (%) Shell thikness Breking strength *p<0.05, **p<0.01, ***p< mg Propolis level. Similr trend, ut not signifint, ws notied in 0 or 50 mg Propolis level. There ws signifint negtive reltionship etween plsm gloulin nd residul feed onsumption in lying hens fed diet ontining 150 mg Propolis. Similr trend, ut not sttistilly signifint, ws notied in remining groups. Both oserved feed onsumption nd feed onversion rtio ws signifintly positive orrelted with RFC in ll tretment groups. Positive reltionship etween egg numer nd RFC ws oserved in ll groups. Tixier-Boihrd et l. (1995) indited tht positive orreltion etween RFC nd egg numer. Signifint positive reltionship etween yolk perentge nd RFC ws oserved in ll tretment groups. Similr result ws otined y Fthi nd Gll (007). They reported tht the yolk perentge ws signifintly positive orrelted with RFC in oth rown nd white egg-type strins. El-Syed nd El-Hkim (1994) lulted signifint nd high positive ssoition etween RFC nd the yolk perent in full-si norml nd dwrf hens. The uthor showed tht the positive orreltion proly reflets the inrese in dry mtter perentge nd energy ontent in the egg when the proportion of yolk inreses. REFERENCES Bonomi, A., F. Morletto nd M. Binhi, Propolis in feeds for lying hens. Avioltur, 54: Buhtel, T., S. Ves, A. Dimitrin nd I. Moldovn, Contriution to knowledge of the stimultive effet of propolis on piglet nd pullets. Buletinul Institului Agronomi Cluj Npo Zoot. Med. Vet., 37: Chndr, R.K. nd P.M. Newerne, Nutrition, immunity nd infetions. Plenum Press, New York. 1110

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