Egg quality, fatty acid composition and immunoglobulin Y content in eggs from laying hens fed full fat camelina or flax seed

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1 Cherin nd Quezd Journl of Animl Science nd Biotechnology (2016) 7:15 DOI /s y RESEARCH Open Access Egg qulity, ftty cid composition nd immunogloulin Y content in eggs from lying hens fed full ft cmelin or flx seed Git Cherin * nd Nthlie Quezd Astrct Bckground: The current study ws conducted to evlute egg qulity nd egg yolk ftty cids nd immunogloulin (IgY) content from lying hens fed full ft cmelin or flx seed. Methods: A totl of 75, 48-week-old Lohmn rown hens were rndomly llocted to 3 tretments, with 5 replictes contining 5 lying hens ech replicte. The hens were fed corn-soyen sl diet (Control), or Control diet with 10 % of full ft cmelin (Cmelin) or flx seed (Flx) for period of 16 wk. Hen production performnce egg qulity, egg yolk lipids, ftty cids nd IgY were determined every 28 d during the experimentl period. Results: Egg production ws higher in hens fed Cmelin nd Flx thn in Control hens (P < 0.05). Egg weight nd lumen weight ws lowest in eggs from hens fed Cmelin (P < 0.05). Shell weight reltive to egg weight (shell weight %), nd shell thickness ws lowest in eggs from hens fed Flx (P < 0.05). No difference ws noted in Hugh unit, yolk:lumen rtio, nd yolk weight. Significnt increse in α-linolenic (18:3 n-3), docospentenoic (22:5 n-3) nd docoshexenoic (22:6 n-3) cids were oserved in egg yolk from hens fed Cmelin nd Flx. Totl n-3 ftty cids constituted 1.19 % in Control eggs compred to 3.12 nd 3.09 % in Cmelin nd Flx eggs, respectively (P < 0.05). Eggs from hens fed Cmelin nd Flx hd the higher IgY concentrtion thn those hens fed Control diet when expressed on mg/g of yolk sis (P < 0.05). Although the egg weight ws significntly lower in Cmelin-fed hens, the totl egg content of IgY ws highest in eggs from hens fed Cmelin (P <0.05). Conclusions: The egg n-3 ftty cid nd IgY enhncing effect of dietry cmelin seed wrrnts further ttention into the potentil of using cmelin s functionl feed ingredient in poultry feeding. Keywords: Cmelin seed, Egg qulity, Flx seed, Immunogloulin Y, n-3 ftty cids Bckground Polyunsturted ftty cids (PUFA) of the omeg-3 (n-3) fmily hve wide rnge of demonstrted helth-relted enefits. These positive effects include: crdioprotective, nticncer, triglyceride nd lood pressure lowering, immune helth enhncing nd their roles in growth nd mturtion of centrl nervous system [1 3]. Dietry n-3 ftty cids include α-linolenic cid (18:3 n-3), eicospentenoic cid (EPA, 20:5 n-3), docospentenoic cid (DPA, 22:5 n-3) nd docoshexenoic cid (22:6 n-3, DHA). α-linolenic cid is present in terrestril oils (e.g. flx) while EPA, DPA, nd DHA re present in mrine * Correspondence: git.cherin@oregonstte.edu Deprtment of Animl nd Rngelnd Sciences, Oregon Stte University, Corvllis, OR 97331, USA oils. Due to the limited consumption of n-3 ftty cidrich mrine foods (e.g. ftty fish) in western countries, there is n incresed interest in modifying niml food products with n-3 ftty cids [4, 5]. In this context, studies on enriching poultry foods (e.g. eggs, met) with n-3 ftty cids through feeding strtegies re topic of continued interest [6, 7]. Oil seeds such s flx re usully incorported into poultry diets due to their nutritionl vlue such s metolizle energy nd crude protein content. Flxseed contins out 34 % oil nd high content of α- linolenic cid (>50 %) mkes it common feed ingredient for n-3 ftty cid enrichment [8, 9]. Due to the cost nd limited vilility of feed ingredients rich in n-3 ftty cids, lternte novel n-3 feedstuffs re explored Cherin nd Quezd. Open Access This rticle is distriuted under the terms of the Cretive Commons Attriution 4.0 Interntionl License ( which permits unrestricted use, distriution, nd reproduction in ny medium, provided you give pproprite credit to the originl uthor(s) nd the source, provide link to the Cretive Commons license, nd indicte if chnges were mde. The Cretive Commons Pulic Domin Dediction wiver ( pplies to the dt mde ville in this rticle, unless otherwise stted.

2 Cherin nd Quezd Journl of Animl Science nd Biotechnology (2016) 7:15 Pge 2 of 8 Cmelin stiv or flse flx or wild flx is n oilseed crop of the Brssic (Crucifere) fmily tht contins high levels of n-3 ftty cids [10, 11]. Although cmelin hs een cultivted since the Bronze Age, there is renewed interest in cmelin s feedstock for io-fuel production. Investigtions on the nutritionl vlue in poultry feeding such s metolizle energy [12, 13], digestiility [13, 14], egg nd met n-3 enriching [14 16] nd ntioxidnt properties [17] of cmelin coproducts (e.g. mel, cke) hve een documented. However, no informtion is ville on n-3 ftty cid enriching nd immune-relted effects of feeding full ft seeds of cmelin in poultry. Considering the high demnd of flx for humn helth-food uses, finding lternte sources of n-3 ftty cid-rich feeds will reduce production costs nd will provide n-3 PUFA-enriched foods for humn consumption. In this context, the ojectives of the current study were to investigte the effect of feeding full ft cmelin seeds to lying hens on egg qulity, lipid nd ftty cid composition, nd egg production, during 4 mo period of feeding tril. It ws hypothesized tht feeding cmelin seeds will enhnce n-3 ftty cid incorportion in eggs without ffecting egg qulity or hen production spects. In ddition, egg immunogloulin Y (IgY) content ws lso determined, s our previous studies hve shown tht feeding linseed or fish oil rich in n-3 ftty cids led to significnt increse in egg yolk IgY [18]. Chicken egg hs een extensively studied s n importnt source of commercil ntiodies nd investigtions on hen diet modultion to produce IgY my provide novel vlue-dded nutrceuticls from eggs. Methods An institutionl niml cre nd use committee pproved ll experimentl protocols to ensure dherence to Animl Cre Guidelines. Birds, diets, nd housing The cmelin nd flx seed used in the current study were nlyzed for gross energy, crude protein, mino cids, minerls nd crude ft content t the University of Arknss Poultry Science Centrl Anlyticl Lortory nd for ftty cids t Oregon Stte University (Tle 1). A totl of 75, 48-week-old rown lying hens (Lohmn Brown) were rndomly llocted to 3 tretment groups, with 5 replictes contining 5 lying hens ech replicte. The irds were kept in individul cges (18 in 21 in 23 in) (width length height). The hens were fed cornsoyen sl diet (Control, negtive control), or Control diet with 10 % of full ft cmelin seeds (Cmelin) or flx seed (Flx, positive control) (Tle 2). All the diets were isocloric nd isonitrogenous. The hens originted from study investigting the use of oil seeds in egg lying hens nd were fed diets contining corn-soy-sed control or Tle 1 Chemicl composition nd nutrient profile of full ft cmelin nd flxseed Nutrient composition Cmelin seed Flx seed Gross Energy, kcl/kg 6,090 6,530 Crude protein, % Crude ft, % Amino cid, % Asprtic cid Threonine Serine Glutmic cid Glycine Alnine Vline Isolecucine Leucine Tyrosine Phenyllnine Lysine Tryptophn Histidine Arginine Cystine Methionine Ftty cid, % Plmitic (16:0) Steric (18:0) Oleic cid (18:1) Linoleic (18:2 n-6) α-linolenic (18:3 n-3) Eicosenoic (20:1) Totl Sturted Totl Monounsturted ftty cid Totl n-3 ftty cid Totl n-6 ftty cid Minerls, ppm P 7,027 6,449 K 8,612 7,025 C 2,113 2,022 Mg 3,013 3,439 S 5,827 1,828 N Fe Mn Zn Cu

3 Cherin nd Quezd Journl of Animl Science nd Biotechnology (2016) 7:15 Pge 3 of 8 Tle 2 Experimentl diet composition nd clculted nutrient nlysis Experimentl diets 1 Ingredients, % Control Cmelin Flx Corn grin Soyen mel (49 % CP) Whet middling Limestone Diclcium Phosphte Premix Slt Cmelin seed Flx seed Nutrient composition Metolizle energy, kcl/kg 2,800 2,800 2,800 Crude protein, % Clcium, % Aville phosphorus, % Lysine, % Cystine, % Methionine, % Arginine, % Threonine, % Ftty cid composition, % Plmitic cid (16:0) Plmitolieic cid (16:1) Steric cid (18:0) Oleic cid (18:1) Linoleic cid (18:2 n-6) α-linolenic cid (18:3 n-3) Eicosenoic cid (20:1) Totl sturted ftty cids Totl monounsturted ftty cids Totl n-3 ftty cids Totl n-6 ftty cids Totln-6:n-3 ftty cids Control, Cmelin, nd Flx represent corn-soyen mel sl diet (Control); or sl diets contining Cmelin seed t 10 % (Cmelin) or flx seed t 10 % (Flx). 2 Suppled per l feed: Vit A, 740,000 IU; Vit D ,000 ICU; Vit E 1,200 IU; Vit B 12, 1.6 mg; Rioflvin, 800 mg; Pntothenic cid, 1,000 mg; Nicin, 6,000 mg; Mendione, 135 mg; Choline, 500 mg; Thimine, 135 mg; Folic cid, 45 mg; Pyridoxine,180 mg; d-iotin, 0.15 mg; Ethoxyquin, 2.5 μg; Mngnese, 2.5 %; Zinc, 92.4 mg; Selenium, 120 ppm; Zinc, 2.00 %; Choline, 50,000; Copper sulfte 2,000 ppm; Iodine ppm; Iron 1.8 % cmelin or flx mel for period of four months [12]. Birds were tken off the experimentl diets for 6 wk efore the initition of the current study. Wter nd feed were provided d liitum. The lying hens did not receive ny vccines or drugs during the entire experimentl period. The experimentl diets were fed for period of 16 wk. The irds were mintined on 16 L:8D photoperiod nd stndrd conditions of temperture nd ventiltion s per University Poultry Frm stndrd operting procedures. Assessing Egg production nd Egg qulity Egg production ws recorded on individul hens nd egg production (%) ws clculted s totl eggs divided y the totl numer of dys nd hens. A totl of 10 eggs (2 from ech replicte) from ech tretment were tken every 28 dy to ssess egg qulity prmeters, ftty cid profile, lipid nd IgY content. The eggs were weighed, nd yolks were seprted using n egg seprtor nd were rolled on wet pper to remove ny white lumen nd were then weighed. Two yolks were pooled to otin smple size of 5 for ftty cid nlyses. Alumen weight ws clculted y sutrcting yolk nd shell weight from totl egg weight. Hugh unit (HU) [19] ( mesure of lumen thickness) ws determined y mesurement lumen height y using tripod micrometer. The Hugh units (HU) were clculted y the formul HU = 100 log (H W0.37), where H is the verge lumen height (mm) nd W is the weight of the egg (g). Shell ws wiped clen nd weighed. Shell thickness ws mesured using n electronic micrometer. Yolk color ws mesured y compring yolk color to the Roche yolk color fn. Totl lipid nd ftty cid nlysis Aout 2 g of feed or egg yolk, ws tken for totl lipid extrction using chloroform: methnol (2:1) following the method of Folch et l. [20]. Ftty cid methyl esters were prepred from totl lipid extrct using methnolic HCl [21]. An internl stndrd (23:0) (Mtrey, PA) ws used for ftty cid quntifiction. The nlysis of ftty cids were performed with n Agilent 6890 gs chromtogrph (Agilent Technologies, CA) equipped with n utosmpler, flme-ioniztion detector, nd fused-silic cpillry column, 30 m 0.25 mm 0.2 μm film thickness (Supelco, PA). Ech smple (1 μl) ws injected with helium s crrier gs onto the column progrmmed for incresed oven tempertures (the initil temperture of 110 C ws held for 0.5 min, then incresed y 20 C/min to 190 C, held for 7 min, nd then incresed t 5 C/min to 210 C nd held for 8 min). Inlet nd detector tempertures were oth 250 C. Pek res nd ftty cid percentges were clculted using Agilent ChemSttion softwre (Agilent Technologies, CA). Ftty cid methyl esters were identified y comprison with retention times of uthentic stndrds (Mtrey, PA) nd were expressed s percentges of totl ftty cid methyl esters or s mg/egg.

4 Cherin nd Quezd Journl of Animl Science nd Biotechnology (2016) 7:15 Pge 4 of 8 Determintion of IgY concentrtion in Egg yolk Egg immunogloulin Y ws isolted from egg yolk y the method descried erlier [22]. Briefly, out 150 to 200 mg of pooled egg yolk ws diluted 1:6 (v/v) with cidified deionized wter (ph 2.5), vortexed well, nd stored t 4 C. After overnight refrigertion, smples were centrifuged t 10,062 g t 4 C for 15 min nd the superntnts were collected nd egg IgY contents were quntified y ELISA using rit nti-chicken IgG (Rocklnd Inc., Gilertsville, PA) s descried erlier [23]. Egg weight nd yolk weight were used to clculte IgY in milligrms per grm of egg yolk or totl IgY per egg. Sttisticl nlyses The effects of diet on hen production performnces, egg qulity, egg totl lipids, ftty cid composition nd IgY content were nlyzed y two-wy ANOVA using SAS 9.4 [24]. Diet nd week were the min fctors. Ech cge ws considered s n experimentl unit. Significnt differences mong tretment mens were nlyzed y Tukey s HSD test t P < 0.05 [25]. Results nd discussion The nutrient profiles of the two oil seeds (cmelin nd flx) re reported in Tle 1. The gross energy content of cmelin nd flx seed were 6,090 nd 6,530 kcl/kg. The crude protein content ws higher in cmelin seed (25.8 %) thn flx seed (19.0 %). The crude ft constituted 38.9 % in cmelin seed compred to 42.0 % in flx seed. The protein of cmelin nd flx seed contined severl essentil mino cids such s threonine, methionine, vline, isoleucine, leucine, lysine, nd phenyllnine (Tle 1). The α-linolenic cid (18:3 n-3) in cmelin seed ws comprtively lower thn tht of flx seed (Tle 1). Linoleic cid (18:2 n-6) ws higher in cmelin seed thn flx seed. Cmelin seeds lso hd very high levels of eicosenoic cid (20:1). Totl sturted ftty cids (14:0 + 16:0 + 18:0) constituted 9.04 nd 9.08 % for cmelin nd flx seed, respectively. Plmitic cid (16:0) ws mjor sturted ftty cid followed y steric cid (18:0) in oth oil seeds. The high protein, energy nd n- 3 nd n-6 ftty cid content of cmelin seed mkes it potentilly suitle source of plnt protein nd essentil n-6 nd n-3 ftty cid source in poultry diets. Among the severl minerls in cmelin seed, potssium ws the mjor minerl followed y phosphorus, sulfur, mgnesium nd clcium. The experimentl diet composition, nutrient composition nd ftty cid profile of the diets re shown in Tle 2. Inclusion of cmelin nd flx seed led to increse in α-linolenic cid in the diet long with reduction in sturted ftty cids. With greter proportion of n-3 PUFAs, the proportion of n-6 PUFA decresed in the diets. Thus, the rtio of n-6 PUFAs to n-3 PUFAs verged 7.69, 1.43 nd 0.71 for Control, Cmelin nd Flx, respectively. Oleic cid in the diets constituted 19.9, 18.7 nd 17.5 for Control, Cmelin nd Flx, respectively. Totl monounsturted ftty cids (16:1 + 18:1 + 20:1) were higher in Cmelin thn Flx nd Control due to the presence of eicosenoic cid. Hen production nd Egg qulity A 4 % increse in egg production ws noted in hens fed the Cmelin nd Flx (P < 0.05) (Tle 3). No difference in egg production could e oserved etween hens fed Cmelin nd Flx. A significnt increse feed consumption (g/d) ws oserved in hens fed Cmelin nd Flx compred to Control hens (P < 0.05). Egg weight nd lumen weight ws lowest in eggs from hens fed Cmelin (P < 0.05). Shell weight reltive to egg weight (shell weight %), nd shell thickness ws lowest in eggs from hens fed Flx (P < 0.05). No difference ws noted in Hugh unit, yolk:lumen rtio, nd yolk weight. Reduction in egg nd lumen weight oserved in the current study corroorted with our previous studies on feeding cmelin mel in lying hen diets [12]. Cmelin elongs to the Brssic fmily nd species of this fmily re high in non-strch polyscchrides, glucosinoltes nd other ntinutritionl fctors such s trypsin inhiitor [26]. While ssessing the digestiility in hens fed diets contining cmelin mel we oserved significnt reduction in crude protein digestiility [12]. It is not known if the protein digestiility ws impired in the current study ffecting nitrogen retention nd egg lumen weight. Most studies on cmelin in lyer hen feeding were on coproducts such s mel or cke [14, 26, 27]. To uthor s knowledge, no studies hve shown impct of dietry full ft cmelin seeds on egg chrcteristics. Inclusion of flx seeds in lying hens diets significntly decresed the yolk color (Tle 3). The yellow color of egg yolk is produced y xnthophyll pigments, derived from diet. In this study, we oserved lighter egg yolk color from lying hens fed flx seeds, proly due to lesser pigments in the diets. Inclusion of flx seed led to reduction (~9 %) in the mount of corn compred to Control diets used in the diets consequently ffecting the pigment content nd leding to pler yolk. However, we did not mesure the pigment content in the experimentl diets. Totl lipids nd ftty cid composition of Egg yolk Totl lipid content nd ftty cids profiles of egg yolk of lying hens fed Cmelin nd Flx is presented in Tle 4. No difference ws oserved in the totl ft content of eggs nd ws 33.04, 33.30, nd % for Control, Cmelin nd Flx, respectively (P > 0.05). As the hens, ged significnt increse in egg lipids ws noted.

5 Cherin nd Quezd Journl of Animl Science nd Biotechnology (2016) 7:15 Pge 5 of 8 Tle 3 Effect of dietry inclusion of cmelin nd flx seeds on production performnce of rown lying hens during 16 wk of feeding period Egg prmeters Dietry tretments 1 Pooled P vlue Control Cmelin Flx SEM Diet Week Diet x Week Egg production, % Feed consumption, g/d Egg weight, g Yolk weight, g Shell weight, g Alumen weight, g Shell thickness, mm Yolk weight, % Yolk color Shell weight, % Alumen weight, % Hugh unit Yolk:lumen , Mens within row with no common superscript differ significntly (P < 0.05). Pooled SEM = stndrd error of men 1 Control, Cmelin nd Flx represent corn-soyen mel sl diet (Control); or sl diets contining cmelin (Cmelin) or flx seeds (Flx) t 10 % A significnt diet week interction ws oserved for egg lipids. The ltertion of egg lipids through diet hs long een studied ut still receives lot of ttention ecuse of its implictions for humn helth [28, 29]. Furthermore, consumers re ecoming incresingly interested in purchsing eggs from hens fed plnt-sed feeds contining high mount of n-3 polyunsturted ftty cids. In the current study it ws oserved tht feeding oil seeds such s cmelin nd flx led to significnt chnges in egg yolk ftty cid profile (Tle 4, Fig. 1). Plmitic cid ws the mjor sturted ftty cid in eggs Tle 4 Effect of cmelin nd flx seed in the diet of lyer irds on egg ftty cid composition Dietry Tretments d P-Vlue Egg lipids, % Control Cmelin Flx Pooled SEM Diet Month Diet x Week Plmitic cid (16:0) Plmitoleic cid (16:1) Steric cid (18:0) Oleic cid (18:1) Linoleic cid (18:2 n-6) α-linolenic cid (18:3 n-3) Eicosenoic cid (20:1) Archidonic cid (20:4 n-6) Docospentenoic (22:5 n-3) 0.06 c Docoshexenoic cid (22:6 n-3) Totl sturted ftty cids Totl monounsturted ftty cids Totl omeg-6 ftty cids Totl omeg-3 ftty cids Totl omeg-6:omeg Totl lipids, % ,,c Mens within row with no common superscripts differ significntly (P < 0.05). Pooled SEM = pooled stndrd error of men. d Control, Cmelin nd Flx represent corn-soyen mel sl diet (Control); or sl diets contining cmelin (Cmelin) or flx seeds (Flx) t 10 % Totl sturted ftty cids (14:0 + 16:0 + 18: ); Totl monounsturted ftty cids (16:1 + 18:1 + 20:1 + 22:1); Totl n-6 polyunsturted ftty cids (18:2 n :4 n :3 n :4 n :5 n-6); Totl n-3 polyunsturted ftty cids (18:3 n :5 n :6 n-3)

6 Cherin nd Quezd Journl of Animl Science nd Biotechnology (2016) 7:15 Pge 6 of 8 followed y steric cid. No effect of dietry tretments on egg plmitic cid (P = 0.09) nd totl sturted ftty cids (14:0 + 16:0 + 18:0 + 20:0) ws oserved (P < 0.085). Oleic cid ws lower in eggs from hens fed Cmelin nd Flx when compred to Control (P < 0.05). Eicosenoic (20:1) ws higher in egg yolks from hens fed Cmelin thn Flx nd Control which is not surprising ecuse the cmelin seeds contined 15.7 % of eicosenoic cid. Overll, ddition of oil seeds in the hen diet led to reduction in totl monounsturted ftty cids (16:1 + 18:1 + 20:1) in egg. The inclusion of full ft oil seeds incresed the concentrtion of α-linolenic cid, DPA nd DHA in the egg yolk (P < 0.05). The increse in DPA nd DHA in eggs from hens fed Cmelin nd Flx suggest tht lying hens cn desturte nd elongte α-linolenic cid, the prent n-3 ftty cid precursor to form long chin 22-C n-3 ftty cids. The highest incorportion of DPA ws oserved in eggs from hens fed Cmelin. Docospentenoic cid is usully the mjor long chin n-3 ftty cid in poultry tissues fed plntsed sources with high α-linolenic cid nd the current study is in greement with other reported reserch [30]. The n-3 ftty cid content of eggs from Cmelin in the current study ws lower thn our previous reported reserch on feeding cmelin mel to hens. Full ft seeds of the Brssicc fmily like cmelin re high in cell wll non strch polyscchrides which limit exposure of seed oils to hen digestive enzymes ffecting digestiility nd egg incorportion. No difference ws oserved in the linoleic cid nd totl n-6 ftty cids content of eggs, lthough the oil seed diets hd lower linoleic cid thn Control. However, rchidonic cid nd totl n-6:n-3 ftty cid rtio ws Fig. 1 α-linolenic cid, long chin n-3 ftty cid nd totl n-3 ftty cid supplied through eggs from lyer hens fed Control, Cmelin or Flx. - Mens etween diets nd within ftty cid type without common letter differ significntly (P < 0.05). Control, Cmelin nd Flx represent corn-soyen mel sl diet (Control); or sl diets contining cmelin (Cmelin) or flx seeds (Flx) t 10 % lowest in eggs from Cmelin nd Flx (P < 0.05). The findings of the present study re in greement with our previous reported reserch [14, 21] showing n increse in α-linolenic cid, followed y decresed in rchidonic cid nd n-6:n-3 rtio in eggs from hens fed flx seeds or cmelin mel. The high linolenic cid nd lower rtio of n-6:n-3 ftty cids in Cmelin nd Flx my hve decresed the competition of linoleic with α-linolenic cid for desturse nd elongse enzymes involved in ioconversion to linoleic to rchidonic cid resulting in reduced egg content of rchidonic cid. Consumer interest in food products enriched with functionl nutrients such s n-3 ftty cids re growing rpidly nd egg is suitle vehicle for delivering helthpromoting omeg-3 ftty cids [31]. For exmple, consumption of two eggs (1 serving) from hens fed Cmelin nd Flx could provide over 300 mg of totl n-3 ftty cids with over 150 mg of it eing long chin 22-C n-3 ftty cids (Fig. 1). The verge per cpit intke of long chin n-3 PUFA is pproximtely g per dy in North Americ [32]. The Dietry Guidelines for Americns recommends 0.65 g nd the WHO recommends dily intke of mg of EPA nd DHA [32]. Therefore, consuming two eggs from hens fed Cmelin could provide over 30 to 50 % of the extr needed long chin n-3 PUFA in the diet. The role of dietry flx seeds in enriching egg n-3 ftty cids is well documented [9, 21]. However, to the uthor s knowledge, the effect of feeding full ft cmelin seeds on egg yolk ftty cid composition is not known. Egg immunogloulin Y ImmunogloulinY concentrtion of egg yolk nd totl IgY in the egg is shown in Fig. 2. The hens fed Cmelin nd Flx hd the higher IgY concentrtion thn those hens fed Control diet when expressed on mg/g of yolk sis (P < 0.05). Although the egg weight ws significntly lower in Cmelin-fed hens, the totl egg content of IgY ws highest in eggs from hens fed Cmelin (P < 0.05). Previously we reported significnt increse in hen serum IgG nd egg IgY in hens fed diets high in n-3 ftty cids such s linseed or fish oil when compred to sunflower oil or niml tllow [18]. The rtio of dietry n-6 to n-3 PUFA is decisive fctor in modultion of IgG formtion y immune cells nd susequently trnsporttion in the plsm nd deposition to egg yolk. In the current study dietry n- 6 to n-3 ftty cid rtio ws over 5 to 11-fold higher in Control compred to Cmelin nd Flx. Chnges in dietry n-6 to n-3 ftty cid rtios cn induce significnt ltertions in immune cell PUFA composition nd eicosnoid production in lyer hens [33]. Our previously reported reserch hs shown tht high dietry n-3 ftty cids cn led to decresed production of rchidonic cid-derived eicosnoids such s prostglndin E2 s well s interleukins

7 Cherin nd Quezd Journl of Animl Science nd Biotechnology (2016) 7:15 Pge 7 of 8 Acknowledgments The uthors wish to cknowledge support from the Oregon Stte University Agriculture Reserch Foundtion wrd to G. Cherin. The cmelin seed used in this study ws kindly supplied y Willmette Biomss Processors, Inc, Rickrell, OR. Received: 24 August 2015 Accepted: 22 Ferury 2016 Fig. 2 Effect of cmelin nd flx seed in the diet of lyer irds on egg immunoglouliny (IgY) content. - Mens etween diets without common letter differ significntly (P < 0.05). Control, Cmelin nd Flx represent corn-soyen mel sl diet (Control); or sl diets contining cmelin (Cmelin) or flx seeds (Flx) t 10 % (IL-6), which cn ffect the production of immunogloulins [34, 35]. No informtion on feeding full ft cmelin seeds on egg yolk IgY is known. Over the lst decde, use of chicken eggs for hrvesting ntiodies for therpeutic use hs een ttempted. In this context, use of diet in modulting egg IgY my open new venues for isolting vlue-dded functionl ingredients for phrmceuticl or other nutrceuticl purposes. Conclusions In conclusion, the results of the present study indicte tht feeding cmelin or flx seeds enhnced egg production while enriching eggs with helth-promoting n-3 ftty cids. The effect tht feeding cmelin incresing IgY in egg wrrnts further ttention into the immunomodulting properties of this oil seed. Cmelin is nonfood crop nd considering the lck of competition of cmelin for humn food uses, its potentil to develop s functionl livestock feed for food enrichment s well s niml helth deserves further reserch. Competing interests The uthors declre tht they hve no competing interests. Authors contriutions GC crried out the experimentl design, reviewed the literture, conducted sttisticl nlyses, drfted nd edited the mnuscript. NQ collected the dt nd conducted chemicl nlysis. All uthors red nd pproved the finl mnuscript. 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