S. Canan Bölükbası, M. Kuddusi Erhan and Hilal» Urüsan

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1 doi: +.,+.+/jps.31, Copyright,+, Jpn Poultry Science Assocition. S. Cnn Bölüksı, M. Kuddusi Erhn nd Hill» Urüsn Atturk University, Fculty of Agriculture, Deprtment of Animl Science, Erzurum, Turkey This reserch ws conducted to determine ects of dietry ergmot oil levels (,,/., /., 1/. m l/kg) on performnce, egg qulity, lood metolic profile nd ftty cid composition of egg yolk in lying hens. Sixty four of 01 weeks old white Lohmn LSL lying hens were rndomly ssigned to four groups eqully ( n +0). Ech tretment ws replicted four times Dietry supplementtion of ergmot oil hd no significnt ect on feed conversion rtio, egg weight, nd egg production, shell thickness, rtio of lumen nd shell. But, supplementtion of ergmot oil decresed feed intke. The ddition of /. m l/kg ergmot oil to the lying hens feed led to significnt increse in the yolk rtio. It ws lso oserved tht egg shell rtio, serum cholesterol nd clcium concentrtion reduced significntly with supplementtion of ergmot oil in lying hens diets. The highest IgG concentrtion ws otined from hens fed,/. m l/kg ergmot oil. Addition of ergmot oil to feeds significntly incresed eicospentenoic cid (EPA), docoshexenoic cid (DHA) nd n-- concentrtion nd decresed n- 0/n-- rtio in egg yolk. The results of this reserch indicted tht the ddition of ergmot oil to the lying hens feed led to significnt decrese in the feed intke nd concentrtions of serum cholesterol. It ws lso concluded tht dietry supplementtion of ergmot oil significntly incresed egg shell strength nd, the EPA, DHA nd n-- rtio of the egg yolk. Key words: ergmot oil, cholesterol, egg production, ftty cid, lying hen J. Poult. Sci.,.1: ,,+ Introduction mtory, nti-dirrhoe properties) nd ctivity in di# erent ody systems, e.g. endocrine nd immune system. Growing concern out ntiiotic growth promoters in Some reserchers reported tht plnt extrcts stimulted niml nutrition hs creted tremendous orts to develop the voluntry feed intke especilly in young nimls nd use di# erent plnt compounds s possile nturl (Rieu et l., +331 ; Ultee et l.,,,; Ginnens et l., lterntives. Phytogenic feed dditives or plnt extrcts,-; Kroismyr et l.,,2). lso referred to s Phytogenics - re n extremely heterogeneous Rmkrishn et l. (,-) nd Willims nd Los (,+) group of feed dditives originting from leves, ssumed tht phytogenics my stimulte the production of roots, tuers or fruits of hers, spices or other plnts. digestive enzymes such s lipse nd mylse, thus hving They re either ville in solid, dried nd ground form eneficil ect on nutrient utiliztion in di# erent cteor s extrcts or essentil oils.(steiner,,3). gories of nimls. Essentil oils hve een known iologicl ctivity, in- It hs een reported severl plnt extrcts hd ntiterolemics cluding nticteril, ntivirucidl, ntifungl, hypocholes- microil, nticoccidil, fungicidl or ntioxidnt properties nd nti-inflmmtory ects (Cown, +333 ; in vitro (Cown, +333 ; Fleiro et l.,,-). Some studies Crig, +333 ; Fleiro et l.,,-). Recoqilly (,0) repthogens, hve shown tht essentil oils of rosemry ( Rosmrinus ported tht plnt extrcts nd their essentil oils hve o$ cinlis), sge ( Slvi sclre), thyme ( Thymus vulgris) wide rnge of ctivities, including inhiitory ction on in this respect were the most ctive ginst strins of ects on physio-pthologies (e.g. nti-inflm- E. coli (Smith-Plmer et l., +332 ; Hmmer et l., +333; Dormn nd Dens,,). Jmroz et l. (,-) deter- Received: August +.,,3, Accepted: Decemer.,,3 Relesed Online Advnce Puliction: Jnury,/,,+ mined tht plnt extrct (crvcrol, cinnmldehyde nd Correspondence: S. Cnn Bölük ı, Attürk University, Agriculturl cpsicin) reduced the totl E. coli nd Clostridium Fculty, Animl Science Deprtment,,/,. Erzurum, Turkey. perfringes numers in the intestines of roiler chickens. (E-mil: cnnoluksi@hotmil.com) Bergmot ( Citrus ergmi), n romtic her, is

2 164 J. Poult. Sci.,.1 (,) memer of the Rutcee. The essentil oil of ergmot oil Tle +. Composition of diets o# ered to lying hens is extrcted from the peel of the fruit y cold-pressing procedure or stem distilltion. Bergmot essentil oil is Ingredients Composition (g/kg) composed of vrious chemicl constituents nd includes Corn /-/40- -pinene, -pinene, myrcene, limonene (. ), -erg- terpineol (Cum et l., +33+ ). Kırslr et l. (,+) indi- Soyen mel..,/14.. Whet 24/0 ptene, -isolene, linlool ( 2 ), linlyl cette (,2 ), Limestone 342 nerol, neryl cette, gerniol, gerniol cette nd - Soyen oil +/4- Diclcium phosphte ++4/. cted tht the chemicl composition of the ergmot oil + Vit Min premix, ws stongly influenced y hrvesting time. It ws de- Slt / clerded tht ergmot, nd its mjor components citrl, DL-methionine 33 +4,- limonene nd linlool, hve een generlly recognised s sfe (Food nd Drug Administrtion,,/). L-Lysine Totl 4/ + It hs een reported tht ergmot oil exhiited nlgesic, ntidepressnt, ntifungl, ntiseptic, ntiiotic, nti- Crude Protein +04, Clculted composition spsmodic, clmtive (Fisher nd Phillips,,0; Snguinetti et l.,,1). Fisher nd Phillips (,0) nd Dens nd Ritchie ( +321) shown tht citrus essentil oils (ergmot, lemon, ME, kcl/kg Methionine Lysine Met nd Cys,1/ , ornge etc) hve potentil ctericidl properties not only + Supplied per kilogrm of diet: +, IU vitmin A;, / IU ginst yest, moulds nd spore forming cteri ut lso vitmin D -; - IU vitmin E;. mg vitmin K -; - mg vitmin B + ; ginst food-poisoning cteri. There re few pulished dt on the ects of essentil 0mg vitmin B,; - mg nicin; + mg clcium D-pntothente; / mg vitmin B 0 ; 4+/ mg vitmin B +, ; + mg folic cid; 4/ oils components in lying hens. Also, there is no study mg D-iotin; / mg vitmin C; - mg choline chloride; 2 mg known out the ect of ergmot oil on the egg prmeters nd lying hens performnce in the chickens. The mngnese; 0 mg iron; 0 mg zinc; / mg copper; 4/ mg colt;, mg iodine; 4+/ mg selenium. ojective of this study ws to reserch the ects of ergmot oil on the lying performnce, egg qulity, egg yolk ftty cid composition nd metolic profile of lying ugtion t. xg for + minute t,. AST, ALT, hens. triglyceride, VLDL, totl protein, lumin, phosphorus, Mteril nd Methods Experimentl Design nd Animls cholesterol, clcium nd IgG were determined in serum y spectrophotometric metod using commericl kits (Roche) in uto-nlyzer (Cos 0, Jpn). A totl of 0. Lohmn LSL lying hens ged 01 weeks were ssigned to. tretments in completely rndomized Ftyy Acid Anlysis Ftty cid nlyses were performed t the Biotechnolodesign. Four lyers were housed in one cge (/.0.0 gy Appliction nd Reserch Centre. After extrcting cm) with four nimls. The dietrt tretments were: (Folch et l., +3/1) yolk smples were methlted for gs m l/kg ergmot oil (sl diet- Tle + ),.,/,./ or chromtogrphic nlysis (GC- Agilent 032 Mss, 1/. m l/kg ergmote oil. The ergmot oil contin, -- fused silic cpillry column, nd film thickness of,/. limonene, +, linlool,,. linlyl cette (ergmot oil mm). Oven temperture ws from +0/ to, t / / ws purchsed from Ege Lokmn Sn. Tic. Compny, min. Detector temperture ws, ; hed pressure ws Mnis, Turkey). During the experiment ( 2 week) hens / psi. were provided feed nd wter d liitum. Egg production Di# erences etween groups were nlysed with one-wy nd feed consumption were recorded dily. Egg weight nlysis of vrince (ANOVA) y using the sttisticl ws meusered iweekly. Feed conversion rte ws ex- pckge SPSS for Windows ( +333 ), version +.. Tretpressed s kilogrm feed intk eper kilogrm egg produc- ment mens were evluted for sttisticlly significnt tion. Eggs used for evlution of yolks nd lumen were di# erences using Tukey test. roken nd the weights of yolks, lumen, shell, shell strength nd shell thickness recorded ( n 2). At the end of the experiment, lood smples were tken ech tret- Results Feed intke, feed conversion rtio, egg production nd ment ( n 2) in order to determine some lood meto- egg weight of the lyers fed diet contining ergmot oil lites. Also, smple of four eggs ws rndomly collected re presented in Tle,. Throughout the experimentl from ech experimentl group to sses ftyy cid compo- period, the feed intke ws significntly ( P /. ) desition of egg yolk t the end of experiment. cresed in lyers fed diets contining ergmot oil supplementtion Collection of Blood Smples compred to tht of the control. The diet Blood smples were tken from wing ven into dditive- with 1/. m l/kg ergmot oil hd induced decreses in free lood tues. Serum ws otined followinge centrif- feed intke y +-./. There were no ects of the ex-

3 Bölük ı et l. : Bergmot Oil Supplementtion in Lying Hens Diet 165 Tle,. E# ects of dietry ergmot oil on the productive performnce of lying hens Groups Feed Feed conversion Egg production Egg intke (g) rtio (g: g) ( ) Weight (g) ++/4/,412,4/1 4/ 014, Bergmot oil 4,/ m l/kg ++,4. +4/,,4.. 4/ 14, Bergmot oil 4/ m l/kg c +, ,4-/ ,4+ Bergmot oil 41/ m l/kg c ,4, , +43 P : P 4/, : not significnt,,, c: Column mens with no common superscript di# er significntly. Tle -. Influence of dietry Bergmot oil on egg qulity of lying hen Groups Yolk ( ) Alumen ( ) Shell ( ) Hugh Unit Shell thickness Breking strength ( mm) - kg mc, / +4, 4., 1,4/ -4, Bergmot oil 4,/ m l/kg, / 4./ , 124/, / Bergmot oil 4/ m l/kg -, / , 2-4/,4-.-0, Bergmot oil 41/ m l/kg , 4,0 2041,4, ,0 42 P : P 4/, : P 4+, : not significnt,,, c: Column mens with no common superscript di# er significntly. Tle.. Influence of dietry Bergmot oil on metolic profil of lying hen Prmeter Bergmot oil Bergmot oil Bergmot oil 4,/ m/kg l 4/ m/kg l 41/ m/kg l P AST U/L / ,2 ALT U/L, , Triglyceride mg/dl +-4--,.4/ ,.400, Cholesterol mg/d l , ,40 3,400 /4- VLDL mg/dl , ,04 +,4/ Totl protein g/dl /410 4+, /4/0 4+, 04-4,- /4/0 4+2 Alumin g/dl,4,- 42,4,- 40,4, 4+,4, 43 C mg/d l,, / , P mg/dl /4, 4-. /40 42 /4, 4/. IgG g/ l , 4. +4/ / 4/ VLDL very low density lipoprotein. perimentl diets on feed conversion rtio, egg production concentrtion. Serum cholesterol concentrtion for hens nd egg weight. fed /. nd 1/. m l/kg ergmot oil ws lower thn hens The ect of experimentl diets of on egg qulity prm- fed sl diet nd.,/ m l/kg ergmot oil. Bergmot oil eters were shown in Tle -. The ddition of ergmot oil supplementtion decresed serum clcium (C) concento the diet hd no impct on lumen percentge of egg, trtion ( P /. ). The diet with /. m l/kg ergmot oil eggshell thickness nd Hugh unit in this study. The yolk hd induced decreses in C concentrtion y /1. mg/dl percentge of egg ws significntly ( P /. ) influenced compred to control group. IgG concentrtion ws signifisignificntly y tretment. The group fed with the /. m l/kg erg- cntly ( P /. ) influenced y tretment. There were mot oil showed the highest yolk percentge of egg. Shell increse in IgG concentrtion with supple- percentge of egg ws significntly influenced y tret- menttion,/. m l/kg ergmot oil. ment. The ddition of ergmot oil to the sl diet Tle / shows the ect of dietry fctors on egg yolk significntly ( P +. ) reduced shell percentge of egg ftty cid composition. None of the dietry fctors # ectcompred to control group. Shell reking strength in- ed plmitic, plmitoleic, steric, oleic, linoleic, linolenic, cresed dependent on incresing of dietry ergmot oil rchidonic, totl sturted ftty cid (SFA), mono- (1/. m l/kg). unsturted ftty cid (MUFA), polyunsturted ftty None of the dietry fctors # ected serum sprtte cid (PUFA) nd n-0 concentrtions in egg yolk in the trnsminse (AST) lnine trnsminse (ALT), tri- present study. glyceride, VLDL, totl protein, lumin nd phosphorus The groups fed on the /. nd 1/. m l/kg ergmot oil

4 166 J. Poult. Sci.,.1 (,) Tle /. Influence of dietry ergmot oil on ftty cid composition of egg yolk lipids of lying hen Ftty cids Bergmot oil Bergmot oil Bergmot oil ( ) 4,/ m/kg l 4/ m/kg l 41/ m/kg l Plmitic,,411 4,3, ,/43 4.0,,4,+ 4/2 Plmitoleic -4 43,40 4,+,41/ 4+,4.2 4+/ Steric 34,/ ,+ +-4/2 4,/ ,- Oleic , / 4. Linoleic / Linolenic 4+/ , 4,0 4-4,, 4- Archidonic 4-/ 41 4, 40 4,3 4, EPA,4, 43, ,420 4+,,42, 42 DHA 42 4, +4-, SFA -, ,4/. 4+. MUFA., ,4, +4/ // PUFA,,4 +41,, , ,,4+0 n-- -4+/ /.4/+ 4,1.4, 4,/ n ,1 +4-/ +/ / +4/. n- 0/n-- / , -4/, 4+0.4,0 4,2 : P 4/, : P 4+, : not significnt,,, c: Column mens with no common superscript di# er significntly. P diet hd the gretest the proportions of EPA thn other with the present findings, Bölüksı et l. (,3) showed groups. The concentrtions of DHA nd n-- in egg yolk tht supplementtion of dietry essentil oil hd no ect lipids ws significntly ( P /. ) incresed levels of die- on feed e$ ciency, egg weight nd egg production. try ergmot oil. Supplementl ergmot oil tended to There ws significntly increse in yolk percentge of decrese n- 0/n-- rtio in egg yolk of lying hens. The egg with supplementtion./ m l/kg ergmot oil. Similowest proportions of n- 0/n-- in yolk lipids ws otined lrly, Denli et l. (,.) reported tht ddition of + g/kg with the diet contining /. nd 1/. m l/kg ergmot oil. Nigell Stiv extrcts in diets of lying quils incresed Discussion weight of yolk in quil egg. In contrst Bölüksı et l. (,2) reported tht ddition of essentil oils( thyme, It ws reported tht essentil oils hve stimulting sge, rosemry) to diet significntly reduced proportion of ect on niml digestive systems (Lnghout,,; Wil- yolk compred with the control. However, Bölüksı et lims nd Los,,+; Rmkrishn et l.,,-). They l. (,3) found tht Nigell Stiv oil ( +,, nd - m l/kg) postulted tht these ects could e due to the incresed supplementtion in lyer diets hd no impct on yolk production of digestive enzymes nd the improved utiliz- percentge. tion of digestive products through enhnced liver functions. The ddition of ergmot oil to the diet cused The results of this experiment showed tht supplement- significnt decrese percentge of egg shell in this study. tion of the diet with ergmot oil hd no ect on feed In contrst this study, Bölüksı et l. (,2) reported conversion, egg production nd egg weight compred with tht supplementing thyme, sge nd rosemry oil to diet of controls, ut cused significnt decrese feed intke. As lying hen incresed proportion of egg shell ove the result, ddition of ergmot oil directly ected some of control. Similrly, Denli et l. (,.) found tht ddition the performnce indices such s reducing feed consump- of + g/kg lck seed extrct incresed weight of egg shell tion ut no significnt di# erences in the others in this in quil egg nd they reported tht lck seed extrct present study. Di# erent essentil oils cted di# erently to could e ssocited with the included components such s performnce of lying hens the controversil results could thymocinene nd crvcrol. Egg shell strength for hens e due to the ctive sustnces of these oils. However, fed 1/. m l/kg ergmot oil ws highest thn other groups. Hertrmpf (,+) reported tht essentil oils derived In the present experiment AST, ALT, triglyceride, from spices nd hers could e successfully used s growth VLDL, totl protein, lumin nd phosphorus for concenpromoters, since they incresed the feed intke due to trtion were not # ected y supplementtion of ergmot their romtic chrcteristics in chickens. In contrst to oil. Serum cholesterol concentrtion reduce y the sup- our results, some reserchers reported tht supplement- plementtion of ergmot oil. This lso confirms erlier tion of the diet with essentil oils hs improved egg findings of Bölüksı et l. (,1,,2,,3), Cse et l. production, feed e$ ciency, egg weight (El-Sheikh et l., ( +33/ ) nd Elson ( +330 ). The hypocholesterolemic ects +332 ; Akhtr et l.,,-; Denli et l.,,.; Botsoglou of essentil oil re medited y down-regulting the reguet l.,,/; Nsir et l.,,/; Aydın et l.,,0; Çuk ltory enzyme, --hydroxy---methylglutryl coenzyme A et l.,,0;bölüksı et l.,,1,,2). In ccordnce (HMG-CoA) reductse. It ws reported the inhiition of

5 Bölük ı et l. : Bergmot Oil Supplementtion in Lying Hens Diet 167 heptic --hydroxy---methylglutryl coenzyme A (HMG- As to our knowledge, no puliction exists on this topic CoA) reductse ctivity which is key regultory enzyme in literture no comprison could e mde with other in cholesterol synthesis y essentil oils (Crowell, +333). studies. Further work is needed in order to give more In ccordnce with the present findings, Cse et l. ( +33/ ) detiled informtion on this topic in chickens. reported tht inhiition of HMC-CoA reductse lowered In conclusion, supplementtion of di# erent levels of serum cholesterol y, in poultry. It hs een reported ergmot oil directly ected some of the performnce (Qureshi et l., +322) tht when cockerels re fed dietry indices such s reducing feed consumption ut no signifilimonene t levels of,/ + ppm for,0 dys, heptic cnt di# erences in the others. It ws lso determined tht HMG-CoA reductse ctivity nd serum cholesterol show cholesterol concentrtions of serum of lying hens were dose-dependent decrese wheres heptic ftty cid syn- significntly reduced y diet including ergmot oil. The thetse ctivity ws un# ected. ddition of.,/ m l/kg ergmot oil to the lying hens feed The diet with ergmot oil hd induced decreses in C led to significnt increse in the IgG concentrtion in the concentrtion compred to control group. This sitution serum. DHA, EPA nd n-- rtio of the egg yolk incresed might e depend on the reduction in feed consumption. wheres the proportions of n- 0/n-- of the egg yolk de- But, supplementtion of 1/. m l/kg ergmot oil in sl cresed dependent on incresing of dietry ergmot oil diet did not # ect eggshell strength. Occhiuto nd Circost levels. It suggested tht it should e use 1/. m l/kg ergmot ( +330, +331) explned tht ergmottin, nother importnt oil level to increse egg shell strength nd EPA, DHA component of the non-voltile frction of ergmot nd n-- concentrtions in egg yolk in during lte lying, esentil oil, might e endowed with C ntgonist pro- period. perties in vitro. Also it is reported tht ergmottine hs ility to inhiit clcium overlod (Occhiuto nd Circost, +330). It is reported tht plnt extrcts nd their essentil oils hve wide rnge of ctivities, including include the stimultion of ppetite nd feed intke, the improvement of endogenous digestive enzyme secretion nd the ctiv- tion of immune responses nd ntioxidnt ctions (Brtt et l., +332 ; Jmroz et l.,,-,,/; Botsoglou et l.,,/). In this study,.,/ m l/kg ergmot oil diet hd significntly higher IgG concentrtion of serum thn tht of the other groups. It is known tht mny essentil oils hve remrkle ility to oth support the immune system nd increse one s rte of heling (Recoqilly,,0). The ddition of ergmot oil to feed significntly incresed n- 0/n-- rtio in the egg yolk in this reserch. cresed the DHA, EPA nd n-- concentrtion nd de- Dens et l. ( +33- ) nd Recsn et l. ( +331) reported tht dietry supplementtion with plnt essentil oils protects nd mintins levels of PUFA in cell memrnes. Morever, Youdim nd Dens (,) found tht ddition of thyme oil nd thymol in diets of rt chnged in ftty cid composition in the rin phospholipid frction. And they reported tht thyme oil nd thymol provided eneficil ects on the ntioxidnt sttus of the rt rin, which my in turn hve influenced the concentrtions of PUFA, especilly DHA. Therefore the findings of the present study my lso show tht ergmot oil provided eneficil ects on the ntioxidnt sttus of the egg yolk, resulting in concentrtions of PUFA, especilly DHA nd EPA. In contrst our study, it ws reported tht rosemry extrct supplementtion in lyer diets hd no ect on ftty cid composition of egg yolk of lying hen (Glort et l.,,+). In study conducted in roilers, it ws shown tht thyme essentil oil cused lower concentrtions of SFA nd PUFA in the leg nd rest tissues, wheres it cused higher MUFA concentrtions (Bölüksı et l.,,0). References Akhtr MS, Nsır Z nd Aid AR. E# ect of feeding powdered Nigell stiv L. seeds on poultry egg production nd their suitility for humn consumption. Veterinrski-Arhiv, 1-: ,-. Aydin R, Bl MA,» Ozuğur AK, Toprk HHC, Kmlk A nd Krmn M. E# ects of lck seed ( Nigell stiv L.) supplementtion on feed e$ ciency, egg yield prmeters nd shell qulity in chickens. Pkistn Journl of Biologicl Science, 3 :,.-,.1.,0. Brtt MT, Dormn JD, Dens SG, Figueiredo AC, Brroso JG nd Ruerto G. Antimicroil nd ntioxidnt properties of some commercil essentil oils. Flvour nd Frgrnce Journl, +- :,-/, Botsoglou NA, Florou-Pneri P, Botsoglou E, Dtos V, Ginnens I, Koidis A nd Mitrkos P. The ect of feeding rosemry, oregno, s# ron nd -tocopheryl cette on hen performnce nd oxidtive stility of eggs. South Africn Journl of Animl Science, -/: +.- +/+.,/. Bölüksı SC, Erhn MK nd» Ozkn A. E# ect of dietry thyme oil nd vitmin E on growth, lipid oxidtion, met ftty cid composition nd serum lipoproteins of roilers. South Africn Journl of Animl Science, -0: ,0. Bölüksı SC, Erhn MK nd Kynr» O. E# ect of Dietry Thyme Oil on Lying Hens Performnce, Cholesterol Rtio of Egg Yolk nd Escherichi coli Concentrtion in Feces. - rd Joint Meeting of the Network of Universities nd Reserch Institutions of Animl Science of the South Estern Europen Countries, Thessloniki + +, Ferury.,1. Bölüksı SC, Erhn MK nd Kynr» O. The ect of feeding thyme, sge nd rosemry oil on lying hen performnce, cholesterol nd some proteins rtio of egg yolk nd Escherichi Coli count in feces. Archives für Geflügelkunde, 1, :,-+,-1.,2. Bölüksı SC, Kynr» O, Erhn MK nd» Urüsn H. The E# ect of Feeding Nigell Stiv Oil on Lying Hen Performnce, Cholesterol nd Some Proteins Rtio of Egg Yolk nd Escherichi Coli Count in Feces. 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