Interntionl Journl of Poultry Science 7 (12): 1232-1236, 2008 ISSN 1682-8356 sin Network for Scientific Informtion, 2008 Effect of Different Dietry Energy Sources on Induction of Ftty Liver-Hemorrhgic Syndrome in Lying Hens Jinwei Zhng, Diwen Chen nd ing Yu niml Nutrition Institute, Sichun griculturl University, Y n, Sichun, 625014, Chin strct: This experiment ws conducted to determine the effect of different dietry energy sources on heptic lipid content nd induction of Ftty Liver-Hemorrhgic Syndrome in lying hens. One hundred nd fifty Lohmnn commercil lyers were divided into three groups of fifty irds ech. ccording to single fctoril rrngement, irds were given one of three experimentl diets with control(group 1, 2.65Mcl/kg),or high energy diet which ws offered with 7% lrd (group 2, 3.06Mcl/kg) nd offered with 26% cornstrch (group 3, 3.00Mcl/kg). High energy diets decresed feed intke nd egg production (p<0.01), verge egg weight of group 3 (cornstrch) ws decresed (p<0.01); In chicks fed the 7% lrd diet or 26% cornstrch d liitum, heptic EE (ether extrct) nd triglyceride were incresed significntly, cornstrch could increse heptic EE, triglyceride nd totl cholesterol more significnt thn lrd. The verge ctivity of LT nd ST of high energy ws incresed to e more thn 2 times thn control tretment of the lst three times lood collection. The LDH of high energy diet ws incresed with no significnt difference of the first three times lood collection. In our experiment, the irds fed the 7% lrd diets hd the lrgest increse in plsm triglyceride, ut there were no significnt tretment effects on plsm triglyceride content y cornstrch ll the experiment period. The verge hemorrhge score of group 2(lrd diet) ws the highest in the three groups. In chicks fed the 7% lrd diet or 26% cornstrch d liitum hd significntly higher MD vlues. In the present study, plsm LT, ST, LDH ctivities nd TC concentrtion were positively correlted with heptic TG concentrtion (p<0.01). The results showed tht mesurement of enzyme ctivities indictive of liver dmge in irds, prticulrly ST, LT, nd plsm TC concentrtion, is vlule tool in the dignosis of ftty liver-hemorrhgic syndrome in flock of lyers. These results suggest tht dietry crohydrte cn induce FLHS more efficiently tht dietry ft, the hens overfeed high ft diets hve significntly more liver hemorrhge thn high crohydrte diets. Key words: Energy sources, ftty liver-hemorrhgic syndrome, crohydrte, ft, lying hen INTRODUCTION Ftty Liver-Hemorrhgic Syndrom (FLHS) is metolic disorder tht occurs widely in poultry flocks nd hs een reported to e the most frequently dignosed condition in commercil lying hens (Squires nd Leeson, 1988). The decrese in egg production nd incresed mortlity ssocited with FLHS cuse considerle economic loss to egg producers. Severl fctors hve een implicted s hving potentil contriutory elements towrds the occurrence of FLHS, including nutrition, environment, genetics, endocrine, toxins etc (Squires nd Leeson, 1988; Wlzem et l., 1993). Despite extensive reserch in ll these res, the etiology of FLHS remins idiopthic nd no definitive prevention nd cure methods for FLHS hve een outlined for use in live irds. Previous experiments hve demonstrted high energy diet cn effectively induce FLHS in the lying hen nd some plsm enzyme ctivities hve een used to dignose FLHS (Diz et l., 1999; Yousefi et l., 2005), ut the reserches out the effect of different dietry energy sources such s crohydrte nd ft on induction of ftty liver-hemorrhgic syndrome in commercil lying hens hve not een descried efore. It hs een suggested tht oxidtive dmge to the cellulr nd orgnelle memrnes of the liver increses the susceptiility of the liver to hemorrhge (Squires nd Leeson, 1988; Schumnn et l., 2003). Some reserches on the mmmls find tht there re two hits in the process of Stetoheptitis, the first hit is the homeostsis of lipid metolism destroyed, heptic net lipid cn proly increse; the second hit is the inflmmtion (Dy nd Jmes, 1998), so we think tht the heptic net lipid deposition is proly the first step in the process of FLHS of the lying hen, nd the reltionship etween the selected plsm enzyme ctivities for the dignosis of FLHS nd heptic lipid must e determined. MTERILS ND METHODS nimls nd diets: This experiment ws conducted ccording to protocols pproved y Sichun griculturl University niml Cre nd Use Committee. One hundred nd fifty Lohmnn lyers (30 weeks of ge, individully cged) were used in this experiment. Hens 1232
were divided into three groups of fifty irds ech. ll irds were given experimentl diets d liitum from ttery cges equipped with seprte nipple drinkers. Three diets were prtilly formulted in ccordnce with the nutrient requirements of poultry (Ntionl Reserch Councile, 1994). The groups were: 1) group 1 (control, 2.65Mcl/kg); 2) group 2 (high ft diet, offered y lrd, 3.06Mcl/kg) ;3) group 3 (high crohydrte diet, offered y cornstrch, 3.00Mcl/kg). The experiment continued for 85 dys. Smple collection nd nlysis: Egg production nd feed intke ws recorded dily nd expressed y whole experiment period per group. The weight of the irds were mesured t the strt nd the end of the 85-d experiment period. From d 30, lood smples were tken from wing vein t 14-d intervl from 9 irds of ech groups which were mrked y symol. The plsm ws seprted y centrifugtion on 2000g for 5 min nd o susequently stored t -20 C until nlysis. The ctivities of plsm LT, ST, LDH, GLU,TG,TC were determined with commercil kits y SHIMDZU CL8000 clinicl chemistry nlyzer, nd MD y spectrophotometer thiorituric cid method (T). ll the mrked irds were humnly killed t the end of the experiment nd the livers were crefully removed, weighed, nd scored for liver hemorrhge y ssigning score from 0 to 3 with 0 indicting no hemorrhges; 1, up to 10 sucpsulr petechil or ecchymotic hemorrhges; 2, more thn 10 sucpsulr petechil or ecchymotic hemorrhges; nd 3, mssive liver hemorrhge (Diz et l., 1999). Liver smples were tken for the determintion of dry mtter (Vcuum Freezing) nd ft content. Sttisticl nlysis: Dte re expressed s the men ± SEM. Dt were nlyzed using the generl liner model procedure of SPSS 11.0. Men vlues were compred y multiple rnge test (Duncn). The level of significnce ws p<0.05. RESULTS Productive performnce: The effect of different dietry energy sources on productive performnce of lying hen were shown in Tle 2, compred with group 1, ody weight of group 2 nd group 3 were incresed significntly (p<0.05); high energy diets decresed Feed intke nd Egg production significntly (p<0.01); compred with group 1 nd group 2, verge egg weight of group 3 (cornstrch) ws decresed (p<0.01); feedegg conversion rte of group 2 ws decresed with no significnt difference. The composition of liver lipid: Composition of liver lipids were shown in tle 3. For chicks fed the 7% lrd diet or 26% cornstrch d liitum, heptic ether extrct nd triglyceride were incresed significntly (p<0.01) compring with sic diet tretment. Totl cholesterol ws incresed with no significnt difference (p>0.05). s cn e seen from the Tle 3, compred with group 1, phospholipid of group 2 ws incresed significntly (p<0.05), there ws no significnt difference etween the group 2 nd group 3. From tle 3, cornstrch could increse heptic EE (ether extrct), triglyceride nd totl cholesterol more significnt thn lrd. Plsm iochemicl indices: Plsm iochemicl indices which were used to dignosis FLHSH were shown in Tle 3. In the present study, plsm LT, ST, LDH, GLU ctivities nd TG,TC concentrtion were determined. In chicks fed the high energy diet d liitum, The verge ctivity of LT nd ST ws incresed to e more thn 2 times thn control tretment of the lst three times lood collection. The LDH of high energy diet ws incresed with no significnt difference of the first three times lood collection, ut verge ctivity of LDH ws more higher thn the other two groups when the chicks were fed the 7% lrd diet. esides these enzymes, plsm TG nd TC were influenced y high energy diets (Tle 4). In our experiment,the irds fed the 7% lrd diets hd the lrgest increse in plsm triglyceride, ut there were no significnt tretment effects on plsm triglyceride content y cornstrch ll the experiment period, nd it ws interesting tht plsm triglyceride content of group 3 ws even lower thn tht of the control tretment ll the experiment period. In the experiment, plsm TC content of group 2 ws incresed significntly (p<0.01) compring with diet tretment of the lst two times lood collection. Liver hemorrhge score: In the experiment, hens consuming lrd nd cornstrch d liitum develop ftty liver (Tle 5). In generl, the hens overfeed high energy diets ecome oese, develop lrge ftty liver nd hve significntly more liver hemorrhge (wlzem, 1993). ut in our experiment, the verge hemorrhge score of group 2 (lrd diet) ws the highest in the three groups. The ftty liver of group 3 hve no significnt hemorrhgic phenomenon. The concentrtion of MD in plsm were influenced y high energy diets (Tle 5). s cn e seen from the tle 5, hemorrhge score nd the MD concentrtion of plsm hve the sme trend. In chicks fed the 7% lrd diet or 26% cornstrch d liitum hd significntly higher MD vlues thn the control tretment. Result showed tht hens fed the lrd hd higher plsm MD vlue nd hd significntly more severe liver hemorrhge scores thn the hens fed on the cornstrch diet. Compred with group 1, the MD concentrtion of group 2 nd group 3 ws incresed significntly (p<0.01). Correltion coefficients: In the present study, plsm LT, ST, LDH ctivities nd TC concentrtion were positively correlted with heptic TG concentrtion (p<0.01), ecuse the heptic EE is most formed with 1233
Tle 1: Composition nd nutrient levels of experimentl diets Group 1 Group 2 Group 3 Ingredients () (Lrd) (Cornstrch) Corn 65.30 56.60 39.50 Soyen mel 24.90 26.60 11.30 Zein Mel - - 13.00 Cornstrch - - 26.00 Lrd - 7.00 - Clcium cronte 7.52 7.52 7.49 Monoclcium phosphte 0.70 0.72 0.80 DL-methionine 0.10 0.10 0.12 Lysine HCL 0.02-0.33 Chloride 0.05 0.05 0.05 Slt 0.37 0.37 0.37 Vitmin premix 0.04 0.04 0.04 Minerl premix 1.00 1.00 1.00 Clculted nlysis CP (%) 16.50 16.50 16.50 ME (Mcl/kg) 2.65 3.06 3.00 Met (sulphur 0.37 (0.65) 0.36 (0.65) 0.40 (0.65) mino cids) Lys (%) 0.84 0.84 0.84 C (%) 3.50 3.50 3.50 P (%) 0.32 0.32 0.32 Supplied per kilogrm of diets: Vitmin, 5000 IU; Vitmin D 3,500 IU; Vitmin E, 5 IU; Vitmin K, 1 IU; Vitmin 1, 1.5 mg, Vitmin 2, 2.5mg, C-pntothente, 2.5mg, nicin cid, 10 mg; pyridoxine,3mg; iotin, 0.1mg; folic cid, 0.25mg; Vitmin 12, 0.005mg. Supplied per kilogrm of diets: MnSO 4. 7H2 O,100mg,; FeSO 4. 7H2 O, 220mg; ZnSO 4. 7H2O, 150mg; CuSO 4. 7H2 O, 20mg; KI, 2mg; N2SeO 3,0.4 mg. Tle 2: Tretment effects on productive performnce of lying hen Lrd Cornstrch ody weight gin (g) 113.33±23.71 185.56±25.12 186.67±39.41 Feed intke (g/d) 121.72±0.00 111.13±0.00 111.47±0.00 verge egg weight (g) 59.35±0.63 59.17±0.50 55.97±0.52 Egg production ( %) 94.06±0.01 82.44±0.11 86.11±0.13 feed-egg conversion rte 2.04 1.89 2.00 Men vlues in row without the sme superscript smll letter re different(p<0.05),those without the sme superscript cpitl letter re significntly different(p<0.01). Tle 3: Tretment effects on the composition of liver lipid Lrd Cornstrch EE% (EE/Fresh liver ) 8.20±0.01 14.10±0.01 15.00±0.01 TG% (TG/Fresh liver) 4.93±0.27 8.72±0.67 9.65±0.45 TC (mg/100g Fresh liver) 389.34±17.05 418.37±24.88 420.52±12.84 PL% (PL/Fresh liver) 2.09±0.11 2.61±0.16 2.28±0.12 Men vlues in row without the sme superscript smll letter re different(p<0.05),those without the sme superscript cpitl letter re significntly different(p<0.01). triglyceride, heptic EE ws positively correlted with heptic TG concentrtion (p<0.01); heptic TG concentrtion lso positively correlted with heptic TG concentrtion (p<0.01). Plsm GLU nd TG concentrtion were not significntly correlted with liver TG concentrtion (p>0.05) DISCUSSION This study ws crried out to investigte the effects of different dietry energy sources on induction of Ftty Liver-Hemorrhgic Syndrome nd liver lipid content in lying hens. Two energy sources diets were used in this experiment. Egg production nd feed intke of hens fed high energy diets were considerly less thn the hens of control group. This result ws consistent with tht of Hrms et l. (2000) nd Yousefi et l. (2005), who reported tht hens fed high energy diet consumed less feed thn hens fed control diet, one of the mjor resons is tht energy content hs key role in the control of food intke (McN nd oormn, 2002). Chnges in dietry energy concentrtion modulte feed efficiency through dependent pthwys. s dietry energy increses, energy needs re stisfied with decresing feed intke. (Plvnik et l., 1997), so, egg production ws limited y decresed feed intke. Hens consuming the diet d liitum or intuted with the diet in quntities equivlent to usul dily energy intke mintined norml rtes of lying, did not ecome oese, nd did not develop liver hemorrhge. Heptic lipidosis ws often referred to s Ftty liver, the etiology of these syndrome hs een proposed to e nutritionl in nture, oserved prcticlly in response to forcefeeding high energy diets (Vn Elswyk et l., 1994). Only few reserchers hve investigted the effect of different energy sources on heptic lipids of lying hens. Our experiment hs showed tht not only crohydrte ut lso ft could increse heptic lipid content. nd triglyceride is the min lipid which deposited in the liver (Wlzem, 1996). The mrked incresing in percentge of triglyceride nd phospholipid is cused y high energy diets nd there is direct reltionship etween triglyceride nd totl ft content in ftty livers of lying hens. In the present study, plsm LT ctivity ws found to e out 2 times higher in hens of group 3 thn in hens of control group, plsm ST ctivity ws incresed with the sme trend. s two importnt trnsminses in heptocytes, these trnsminses could e relesed from the cytolymph when the hptocytes hd een destroyed. So, some trnsminses nd LDH were used to detecting the Hemorrhgic Syndrom, dmnifiction of the heptocytes nd inflmmtion. In our study, from the second lood collection, plsm ST, LT nd LDH ctivity of hens in group 2 ws higher thn group 3. this result ws supported y the liver hemorrhge score nd MD in plsm in this study. Lipid nd glucose re min energy resource of nimls, the first step in lipid metolism is the hydrolysis of the lipid in the cytoplsm to produce glycerol nd ftty cids. The liver hs centrl role in the storge nd distriution within the ody of ll fuels, including glucose, ut the metolism of glucose nd ftty cids is different. Glucose will e oxidized y ll tissues to synthesize TP. The most importnt pthwy which egins the complete oxidtion of glucose is clled glycolysis. nd et oxidtion is the min pthwy of ftty cids metolism. So, when ftty cid deposited in liver, et oxidtion stress would e strengthened, this is 1234
Tle 4: Tretment effects on plsm iochemicl indices LT (U/L) ST (U/L) LDH (U/L) GLU (mmol/l) TG (mmol/l) TC (mmol/l) -------------------------------------------------- The first lood collection ----------------------------------------------- 0.73±0.10 122.14±2.60 144.44±4.30 13.81±0.18 14.99±0.99 3.14±0.16 Cornstrch 0.98±0.10 128.58±3.76 140.44±13.86 14.08±0.19 18.23±1.07 3.87±0.27 lrd 0.81±0.09 129.43±4.30 147.11±7.17 14.18±0.17 13.87±0.67 3.76±0.33 ------------------------------------------------- The second lood collection ------------------------------------------- 0.38±0.11 145.01±5.44 169.91±12.30 15.84±0.41 15.14±1.77 3.27±0.50 Cornstrch 1.84±0.61 147.34±5.23 217.29±22.21 17.50±0.42 23.81±1.36 4.52±0.09 lrd 0.69±0.18 132.59±2.58 184.33±12.76 16.06±0.31 15.06±2.56 3.46±0.43 ------------------------------------------------- The third lood collection ----------------------------------------------- 0.61±0.11 132.40±4.35 162.89±11.54 15.83±0.36 15.19±1.86 4.38±0.31 Cornstrch 1.50±0.34 152.89±22.97 190.78±18.76 16.71±0.26 20.50±1.61 5.27±0.36 lrd 1.21±0.17 135.61±3.37 193.33±13.50 16.13±0.34 14.49±1.12 4.20±0.38 ------------------------------------------------ The fourth lood collection ---------------------------------------------- 0.67±0.13 135.20±5.56 147.50±9.48 C 14.28±0.41 14.93±1.82 2.91±0.30 Cornstrch 1.94±0.29 210.68±21.23 214.56±13.72 17.31±0.40 28.34±2.05 4.83±0.54 lrd 1.58±0.19 165.73±15.19 190.11±22.38 18.93±0.35 13.18±1.65 4.04±0.53 ------------------------------------------------ The fifth lood collection -------------------------------------------------- 0.84±0.17 136.08±5.04 161.78±15.91 15.43±0.35 14.72±1.17 2.28±0.28 Cornstrch 1.97±0.20 211.16±16.55 213.11±15.05 16.60±0.28 25.76±2.41 4.70±0.62 lrd 1.62±0.14 169.87±11.03 162.44±7.91 16.10±0.46 14.22±1.43 4.22±0.45 Men vlues in row without the sme superscript smll letter re different (p<0.05), those without the sme superscript cpitl letter re significntly different (p<0.01). Tle 5: Tretment effects on the liver hemorrhge score nd MD in plsm Lrd Cornstrch hemorrhge score 0.89±0.35 1.78±0.32 1.22±0.32 MD (nmol/l) 4.93±0.27 9.65±0.45 8.72±0.67 Men vlues in row without the sme superscript smll letter re different (p<0.05), those without the sme superscript cpitl letter re significntly different (p<0.01). Tle 6: Correltion coefficients mong heptic TG nd some iochemicl indices Heptic TG Significnt level LT (plsm) 0.611 ** ST (plsm) 0.580 ** LDH (plsm) 0.527 ** GLU (plsm) 0.363 NS TG (plsm) 0.300 NS TC (plsm) 0.508 ** EE (liver) 0.924 ** TC (liver) 0.624 ** PL (liver) 0.433 * NS=not significnt; * p<0.05; ** p<0.01 cuse of oxidtive dmge from n incresed production of free rdicls in heptocytes. MD is lipid per oxidtive products, indirectly reflecting the degree of per oxidtive dmge of the ody y the per oxidtive degree of lipids (Squires nd Wu, 1992; ucl, 1996; hsn et l., 2003; To et l., 2005). So, our result showed tht hens consumed the diet with lrd, the liver hemorrhge score nd MD in plsm were higher thn hens consumed the cornstrch diets. The sme trend could e found within the ST, LT nd LDH ctivity. The results showed tht mesurement of enzyme ctivities indictive of liver dmge in irds, prticulrly ST, LDH, nd GDH, is vlule tool in the dignosis of ftty liver-hemorrhgic syndrome in flock of lyers (Diz et l., 1999). s cn e seen from our result (Tle 6), correltion coefficients mong heptic TG nd some iochemicl indices were first reserched. In the present study, plsm LT, ST, LDH ctivities nd TC concentrtion were positively correlted with heptic TG concentrtion (p<0.01), ut the plsm LDH ctivities of hens in group 3 hs no significnt difference with control group. So,we do not think tht plsm LDH ctivities ws sensitive tool which could e used to dignose the ftty liver-hemorrhgic syndrome in flock of lyers. These results suggest tht dietry crohydrte cn induce FLHS more efficiently tht dietry ft, the hens overfeed high ft diets hve significntly more liver hemorrhge thn high crohydrte diets. CKNOWLEDGEMENT This study is supported y Progrm for Chngjing Scholrs nd Innovtive Reserch Tem in University with grnt No. IRTO555-5, Chin Ministry of Eduction. REFERENCES hsn, H.,. li nd R. li, 2003. Oxygen free rdicls nd systemic utoimmunity. Clin. Exp. Immunol., 131: 398-404. ucl, R., 1996. Lipid nd lipoprotein oxidtion: sic mechnisms nd unresolved questions in vivo. Redox Rep., 2: 291-307. Dy, C. nd O. Jmes, 1998. Stetoheptitis: tle of two "hits". Gstroenterology, 114: 842-845. Diz, G.J., E.J. Squires, R.J. Julin, 1999. The Use of Selected Plsm Enzyme ctivities for the Dignosis of Ftty Liver-Hemorrhgic Syndrome in Lying Hens. vin Dis., 43: 768-773. 1235
Hrms, R.H., G.. Russell nd D.K. Slon, 2000. Performnce of four strins of commercil lyers with mjor chnges in dietry energy. J. ppl. Poult. Res., 9: 535-541. McN, J.M. nd K.N. oormn, 2002. Poultry feedstuffs: supply, composition nd nutrition vlue. C Interntionl, 81: 1681-1693. Plvnik, I., E. Wx, D. Skln nd S. Hurwitz, 1997. The response of roiler chickens nd turkey poults to dietry energy supplied either y ft or crohydrtes. Poult. Sci., 76: 1000-1005. Ntionl Reserch Council, 1994. Nutrient Requirements of Poultry, Ntionl Reserch Council Ntionl cdemy Press Wshington, D.C. 1994. Schumnn,.E., E.J. Squires, S. Leeson nd. Hunter, 2003. Effect of hens fed dietry flxseed with nd without ftty liver supplement on heptic, plsm nd production chrcteristics relevnt to ftty liver hemorrhgic syndrome in lying hens. r. Poult. Sci., 44: 234-244. Squires, E.J. nd S. Leeson, 1988. etiology of ftty liver syndrome in lying hens. r. Vet. J., 144: 602-609. Squires, E.J. nd J. Wu, 1992. Enhnced induction of heptic lipid peroxidtion y ferric nitrilotricette in chickens susceptile to ftty liver rupture. r. Poult. Sci., 33: 329-337. To, X., Z.R. Xu, X.Y. Hn., Y.Z. Wng nd L.H. Zhou, 2005. Effects of fluoride levels on lipid peroxidtion nd ntioxidnt systems of growing/finishing pigs. sin-ust. J. nim. Sci., 18: 552-556. Vn Elswyk, M.E.,.M. Hrgis, J.D. Willims nd P.S. Hrgis, 1994. Dietry menhden oil contriutes to heptic lipidosis in lying hens. Poult. Sci., 73: 653-662. Yousefi, M., M. Shivzd nd I. Sohri-Hghdoost, 2005. Effect of Dietry Fctors on Induction of Ftty Liver-Hemorrhgic Syndrome nd its Dignosis Methods with Use of Serum nd Liver Prmeters in Lying Hens Int. J. Poult. Sci., 4: 568-572. Wlzem, R.L., C. Simon, T. Morishit, L. Lowenstine nd R.J. Hnsen, 1993. Ftty liver hemorrhgic syndrome in hens overfed purified diet. Selected enzyme ctivities nd liver histology in reltion to liver hemorrhge nd reproductive performnce. Poult. Sci., 72: 1479-1491. Wlzem, R.L., 1996. Lipoproteins nd the lying hen: form follows function. Poult. vin iol. Rev.,7:31-64. 1236