Egg quality, fatty acid composition and immunoglobulin Y content in eggs from laying hens fed full fat camelina or flax seed
© Cherian and Quezada. 2016
Received: 24 August 2015
Accepted: 22 February 2016
Published: 3 March 2016
The current study was conducted to evaluate egg quality and egg yolk fatty acids and immunoglobulin (IgY) content from laying hens fed full fat camelina or flax seed.
A total of 75, 48-week-old Lohman brown hens were randomly allocated to 3 treatments, with 5 replicates containing 5 laying hens each replicate. The hens were fed corn-soybean basal diet (Control), or Control diet with 10 % of full fat camelina (Camelina) or flax seed (Flax) for a period of 16 wk. Hen production performance egg quality, egg yolk lipids, fatty acids and IgY were determined every 28 d during the experimental period.
Egg production was higher in hens fed Camelina and Flax than in Control hens (P < 0.05). Egg weight and albumen weight was lowest in eggs from hens fed Camelina (P < 0.05). Shell weight relative to egg weight (shell weight %), and shell thickness was lowest in eggs from hens fed Flax (P < 0.05). No difference was noted in Haugh unit, yolk:albumen ratio, and yolk weight. Significant increase in α-linolenic (18:3 n-3), docosapentaenoic (22:5 n-3) and docoshexaenoic (22:6 n-3) acids were observed in egg yolk from hens fed Camelina and Flax. Total n-3 fatty acids constituted 1.19 % in Control eggs compared to 3.12 and 3.09 % in Camelina and Flax eggs, respectively (P < 0.05). Eggs from hens fed Camelina and Flax had the higher IgY concentration than those hens fed Control diet when expressed on a mg/g of yolk basis (P < 0.05). Although the egg weight was significantly lower in Camelina-fed hens, the total egg content of IgY was highest in eggs from hens fed Camelina (P < 0.05).
The egg n-3 fatty acid and IgY enhancing effect of dietary camelina seed warrants further attention into the potential of using camelina as a functional feed ingredient in poultry feeding.
KeywordsCamelina seed Egg quality Flax seed Immunoglobulin Y n-3 fatty acids
Polyunsaturated fatty acids (PUFA) of the omega-3 (n-3) family have a wide range of demonstrated health-related benefits. These positive effects include: cardioprotective, anticancer, triglyceride and blood pressure lowering, immune health enhancing and their roles in growth and maturation of central nervous system [1–3]. Dietary n-3 fatty acids include α-linolenic acid (18:3 n-3), eicosapentaenoic acid (EPA, 20:5 n-3), docosapentaenoic acid (DPA, 22:5 n-3) and docosahexaenoic acid (22:6 n-3, DHA). α-linolenic acid is present in terrestrial oils (e.g. flax) while EPA, DPA, and DHA are present in marine oils. Due to the limited consumption of n-3 fatty acid-rich marine foods (e.g. fatty fish) in western countries, there is an increased interest in modifying animal food products with n-3 fatty acids [4, 5]. In this context, studies on enriching poultry foods (e.g. eggs, meat) with n-3 fatty acids through feeding strategies are a topic of continued interest [6, 7].
Oil seeds such as flax are usually incorporated into poultry diets due to their nutritional value such as metabolizable energy and crude protein content. Flaxseed contains about 34 % oil and high content of α-linolenic acid (>50 %) makes it a common feed ingredient for n-3 fatty acid enrichment [8, 9]. Due to the cost and limited availability of feed ingredients rich in n-3 fatty acids, alternate novel n-3 feedstuffs are explored. Camelina sativa or “false flax” or “wild flax” is an oilseed crop of the Brassica (Cruciferae) family that contains high levels of n-3 fatty acids [10, 11]. Although camelina has been cultivated since the Bronze Age, there is renewed interest in camelina as a feedstock for bio-fuel production. Investigations on the nutritional value in poultry feeding such as metabolizable energy [12, 13], digestibility [13, 14], egg and meat n-3 enriching [14–16] and antioxidant properties  of camelina coproducts (e.g. meal, cake) have been documented. However, no information is available on n-3 fatty acid enriching and immune-related effects of feeding full fat seeds of camelina in poultry. Considering the high demand of flax for human health-food uses, finding alternate sources of n-3 fatty acid-rich feeds will reduce production costs and will provide n-3 PUFA-enriched foods for human consumption. In this context, the objectives of the current study were to investigate the effect of feeding full fat camelina seeds to laying hens on egg quality, lipid and fatty acid composition, and egg production, during a 4 mo period of feeding trial. It was hypothesized that feeding camelina seeds will enhance n-3 fatty acid incorporation in eggs without affecting egg quality or hen production aspects. In addition, egg immunoglobulin Y (IgY) content was also determined, as our previous studies have shown that feeding linseed or fish oil rich in n-3 fatty acids led to significant increase in egg yolk IgY . Chicken egg has been extensively studied as an important source of commercial antibodies and investigations on hen diet modulation to produce IgY may provide novel value-added nutraceuticals from eggs.
An institutional animal care and use committee approved all experimental protocols to ensure adherence to Animal Care Guidelines.
Birds, diets, and housing
Chemical composition and nutrient profile of full fat camelina and flaxseed
Gross Energy, kcal/kg
Crude protein, %
Crude fat, %
Amino acid, %
Fatty acid, %
Oleic acid (18:1)
Linoleic (18:2 n-6)
α-Linolenic (18:3 n-3)
Total Monounsaturated fatty acid
Total n-3 fatty acid
Total n-6 fatty acid
Experimental diet composition and calculated nutrient analysis
Soybean meal (49 % CP)
Metabolizable energy, kcal/kg
Crude protein, %
Available phosphorus, %
Fatty acid composition, %
Palmitic acid (16:0)
Plamitolieic acid (16:1)
Stearic acid (18:0)
Oleic acid (18:1)
Linoleic acid (18:2 n-6)
α-Linolenic acid (18:3 n-3)
Eicosaenoic acid (20:1)
Total saturated fatty acids
Total monounsaturated fatty acids
Total n-3 fatty acids
Total n-6 fatty acids
Totaln-6:n-3 fatty acids
Assessing Egg production and Egg quality
Egg production was recorded on individual hens and egg production (%) was calculated as total eggs divided by the total number of days and hens. A total of 10 eggs (2 from each replicate) from each treatment were taken every 28 day to assess egg quality parameters, fatty acid profile, lipid and IgY content. The eggs were weighed, and yolks were separated using an egg separator and were rolled on wet paper to remove any white albumen and were then weighed. Two yolks were pooled to obtain a sample size of 5 for fatty acid analyses. Albumen weight was calculated by subtracting yolk and shell weight from total egg weight. Haugh unit (HU)  (a measure of albumen thickness) was determined by measurement albumen height by using a tripod micrometer. The Haugh units (HU) were calculated by the formula HU = 100 log (H + 7.57 − 1.7 W0.37), where H is the average albumen height (mm) and W is the weight of the egg (g). Shell was wiped clean and weighed. Shell thickness was measured using an electronic micrometer. Yolk color was measured by comparing yolk color to the Roche yolk color fan.
Total lipid and fatty acid analysis
About 2 g of feed or egg yolk, was taken for total lipid extraction using chloroform: methanol (2:1) following the method of Folch et al. . Fatty acid methyl esters were prepared from total lipid extract using methanolic HCl . An internal standard (23:0) (Matreya, PA) was used for fatty acid quantification. The analysis of fatty acids were performed with an Agilent 6890 gas chromatograph (Agilent Technologies, CA) equipped with an autosampler, flame-ionization detector, and fused-silica capillary column, 30 m × 0.25 mm × 0.2 μm film thickness (Supelco, PA). Each sample (1 μL) was injected with helium as a carrier gas onto the column programmed for increased oven temperatures (the initial temperature of 110 °C was held for 0.5 min, then increased by 20 °C/min to 190 °C, held for 7 min, and then increased at 5 °C/min to 210 °C and held for 8 min). Inlet and detector temperatures were both 250 °C. Peak areas and fatty acid percentages were calculated using Agilent ChemStation software (Agilent Technologies, CA). Fatty acid methyl esters were identified by comparison with retention times of authentic standards (Matreya, PA) and were expressed as percentages of total fatty acid methyl esters or as mg/egg.
Determination of IgY concentration in Egg yolk
Egg immunoglobulin Y was isolated from egg yolk by the method described earlier . Briefly, about 150 to 200 mg of pooled egg yolk was diluted 1:6 (v/v) with acidified deionized water (pH 2.5), vortexed well, and stored at 4 °C. After overnight refrigeration, samples were centrifuged at 10,062 × g at 4 °C for 15 min and the supernatants were collected and egg IgY contents were quantified by ELISA using rabbit anti-chicken IgG (Rockland Inc., Gilbertsville, PA) as described earlier . Egg weight and yolk weight were used to calculate IgY in milligrams per gram of egg yolk or total IgY per egg.
The effects of diet on hen production performances, egg quality, egg total lipids, fatty acid composition and IgY content were analyzed by two-way ANOVA using SAS 9.4 . Diet and week were the main factors. Each cage was considered as an experimental unit. Significant differences among treatment means were analyzed by Tukey’s HSD test at P < 0.05 .
Results and discussion
The nutrient profiles of the two oil seeds (camelina and flax) are reported in Table 1. The gross energy content of camelina and flax seed were 6,090 and 6,530 kcal/kg. The crude protein content was higher in camelina seed (25.8 %) than flax seed (19.0 %). The crude fat constituted 38.9 % in camelina seed compared to 42.0 % in flax seed. The protein of camelina and flax seed contained several essential amino acids such as threonine, methionine, valine, isoleucine, leucine, lysine, and phenylalanine (Table 1). The α-linolenic acid (18:3 n-3) in camelina seed was comparatively lower than that of flax seed (Table 1). Linoleic acid (18:2 n-6) was higher in camelina seed than flax seed. Camelina seeds also had very high levels of eicosaenoic acid (20:1). Total saturated fatty acids (14:0 + 16:0 + 18:0) constituted 9.04 and 9.08 % for camelina and flax seed, respectively. Palmitic acid (16:0) was major saturated fatty acid followed by stearic acid (18:0) in both oil seeds. The high protein, energy and n-3 and n-6 fatty acid content of camelina seed makes it a potentially suitable source of plant protein and essential n-6 and n-3 fatty acid source in poultry diets. Among the several minerals in camelina seed, potassium was the major mineral followed by phosphorus, sulfur, magnesium and calcium.
The experimental diet composition, nutrient composition and fatty acid profile of the diets are shown in Table 2. Inclusion of camelina and flax seed led to increase in α-linolenic acid in the diet along with a reduction in saturated fatty acids. With a greater proportion of n-3 PUFAs, the proportion of n-6 PUFA decreased in the diets. Thus, the ratio of n-6 PUFAs to n-3 PUFAs averaged 7.69, 1.43 and 0.71 for Control, Camelina and Flax, respectively. Oleic acid in the diets constituted 19.9, 18.7 and 17.5 for Control, Camelina and Flax, respectively. Total monounsaturated fatty acids (16:1 + 18:1 + 20:1) were higher in Camelina than Flax and Control due to the presence of eicosaenoic acid.
Hen production and Egg quality
Effect of dietary inclusion of camelina and flax seeds on production performance of brown laying hens during 16 wk of feeding period
Diet x Week
Egg production, %
Feed consumption, g/d
Egg weight, g
Yolk weight, g
Shell weight, g
Albumen weight, g
Shell thickness, mm
Yolk weight, %
Shell weight, %
Albumen weight, %
Total lipids and fatty acid composition of Egg yolk
Effect of camelina and flax seed in the diet of layer birds on egg fatty acid composition
Egg lipids, %
Diet x Week
Palmitic acid (16:0)
Palmitoleic acid (16:1)
Stearic acid (18:0)
Oleic acid (18:1)
Linoleic acid (18:2 n-6)
α-Linolenic acid (18:3 n-3)
Eicosenoic acid (20:1)
Arachidonic acid (20:4 n-6)
Docosapentaenoic (22:5 n-3)
Docosahexaenoic acid (22:6 n-3)
Total saturated fatty acids
Total monounsaturated fatty acids
Total omega-6 fatty acids
Total omega-3 fatty acids
Total lipids, %
No difference was observed in the linoleic acid and total n-6 fatty acids content of eggs, although the oil seed diets had lower linoleic acid than Control. However, arachidonic acid and total n-6:n-3 fatty acid ratio was lowest in eggs from Camelina and Flax (P < 0.05). The findings of the present study are in agreement with our previous reported research [14, 21] showing an increase in α-linolenic acid, followed by decreased in arachidonic acid and n-6:n-3 ratio in eggs from hens fed flax seeds or camelina meal. The high linolenic acid and lower ratio of n-6:n-3 fatty acids in Camelina and Flax may have decreased the competition of linoleic with α-linolenic acid for deasturase and elongase enzymes involved in bioconversion to linoleic to arachidonic acid resulting in reduced egg content of arachidonic acid.
Consumer interest in food products enriched with functional nutrients such as n-3 fatty acids are growing rapidly and egg is a suitable vehicle for delivering health-promoting omega-3 fatty acids . For example, consumption of two eggs (1 serving) from hens fed Camelina and Flax could provide over 300 mg of total n-3 fatty acids with over 150 mg of it being long chain 22-C n-3 fatty acids (Fig. 1). The average per capita intake of long chain n-3 PUFA is approximately 0.1–0.2 g per day in North America . The Dietary Guidelines for Americans recommends 0.65 g and the WHO recommends daily intake of 300–500 mg of EPA and DHA . Therefore, consuming two eggs from hens fed Camelina could provide over 30 to 50 % of the extra needed long chain n-3 PUFA in the diet. The role of dietary flax seeds in enriching egg n-3 fatty acids is well documented [9, 21]. However, to the author’s knowledge, the effect of feeding full fat camelina seeds on egg yolk fatty acid composition is not known.
Egg immunoglobulin Y
In conclusion, the results of the present study indicate that feeding camelina or flax seeds enhanced egg production while enriching eggs with health-promoting n-3 fatty acids. The effect that feeding camelina increasing IgY in egg warrants further attention into the immunomodualting properties of this oil seed. Camelina is a nonfood crop and considering the lack of competition of camelina for human food uses, its potential to develop as a functional livestock feed for food enrichment as well as animal health deserves further research.
The authors wish to acknowledge support from the Oregon State University Agriculture Research Foundation award to G. Cherian. The camelina seed used in this study was kindly supplied by Willamette Biomass Processors, Inc, Rickreall, OR.
Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
- Jump DB, Depner CM, Tripathy S. Omega-3 fatty acid supplementation and cardiovascular disease. J Lipid Res. 2012;53:2525–45.View ArticlePubMedPubMed CentralGoogle Scholar
- Kromhout D, Geleijnse JM, de Goede J, Oude Griep LM, Mulder BJ, de Boer MJ, Deckers JW, Boersma E, Zock PL, Giltay EJ. n-3 fatty acids, ventricular arrhythmia-related events, and fatal myocardial infarction in postmyocardial infarction patients with diabetes. Diabetes Care. 2011;34:2515–20.
- Yashodhara BM, Umakanth S, Pappachan JM, Bhat SK, Kamath R, Choo BH. Omega-3 fatty acids: a comprehensive review of their role in health and disease. Postgrad Med J. 2009;85:84–90.View ArticlePubMedGoogle Scholar
- Palmquist DL. Omega-3 fatty acids in metabolism, health, and nutrition and for modified animal product foods. Prof Animal Scientist. 2009;25:207–49.View ArticleGoogle Scholar
- Cherian G. Dietary manipulation of poultry to develop value-added functional foods for humans. In: Gupta L, Singhal KK, editors. Animal Nutrition: Advancements in Feeds and Feeding of Livestock. Jodhpur: Agrotech Publishing Academy; 2011. p. 339–56.Google Scholar
- Cherian G. Lipid modification strategies and nutritionally functional poultry foods. Food science and product technology. In: Nakano T, Ozimek L, editors. Food Science and Product Technology. Trivandrum: Research Sign Post; 2002. p. 77–82.Google Scholar
- Rymer C, Givens DI. N-3 fatty acid enrichment of edible tissue of poultry: a review. Lipids. 2005;40:121–30.View ArticlePubMedGoogle Scholar
- Gonzalez-Esquerra R, Leeson S. Alternatives for enrichment of eggs and chicken meat with omega-3 fatty acids. Can J Anim Sci. 2001;81:295–305.View ArticleGoogle Scholar
- Cherian G. Omega-3 fatty acids: Studies in avians. In: De Meester F, Watson RR, editors. Wild-Type Food in Health Promotion and Disease Prevention: The Columbus® Concept. New York City: Humana Press; 2008. p. 169–78.View ArticleGoogle Scholar
- Budin JT, Breene WM, Putnam DH. Some compositional properties of camelina (Camelina sativa L. Crantz) seeds and oils. J Am Oil Chem Soc. 1995;72:309–15.View ArticleGoogle Scholar
- Cherian G. Camelina sativa in poultry diets: opportunities and challenges. Ch. 17. In: Makkar HPS, editor. Biofuel Co-Products as Livestock Feed: Opportunities and Challenges. Rome: Food and Agricultural Organization of the United Nations; 2012. p. 303–10.Google Scholar
- Aziza AE, Panda AK, Quezada N, Cherian G. Apparent metabolizable energy, crude protein and fatty acid digestibility, egg quality and fatty acid composition of brown laying hens fed camelina or flaxseed meal. J Appl Poult Res. 2013;22:832–41.View ArticleGoogle Scholar
- Pekel AY, Kim JI, Chapple C, Adeola O. Nutritional characteristics of camelina meal for 3-week-old broiler chickens. Poult Sci. 2015;94:371–8.View ArticlePubMedGoogle Scholar
- Cherian G, Campbell A, Parker T. Egg quality and lipid composition of eggs from hens fed camelina sativa. J Appl Poult Res. 2009;18:143–50.View ArticleGoogle Scholar
- Aziza AE, Quezada N, Cherian G. Feeding Camelina sativa meal to meat-type chickens: effect on production performance and tissue fatty acid composition. J Appl Poult Res. 2010;19:157–68.View ArticleGoogle Scholar
- Thacker P, Widyaratne G. Effects of expeller pressed camelina meal and/or canola meal on digestibility, performance and fatty acid composition of broiler chickens fed wheat–soybean meal-based diets. Arch Anim Nutr. 2012;66:402–15.View ArticlePubMedGoogle Scholar
- Aziza AE, Quezada N, Cherian G. Antioxidative effect of dietary Camelina meal in fresh, stored or cooked broiler chicken meat. Poult Sci. 2012;89:2711–8.View ArticleGoogle Scholar
- Wang YW, Cherian G, Sunwoo HH, Sim JS. Dietary polyunsaturated fatty acids significantly affect laying hen lymphocyte proliferation and immunoglobulin G concentration in serum and egg yolk. Can J Anim Sci. 2000;80:597–604.View ArticleGoogle Scholar
- Haugh RR. The Haugh unit for measuring egg quality. US Egg Poultry Magazine. 1937;43:522–55.Google Scholar
- Folch J, Lees M, Sloane-Stanley GH. A simple method for the isolation and purification of total lipids from animal tissues. J Biol Chem. 1957;226:497–509.PubMedGoogle Scholar
- Hayat Z, Cherian G, Pasha TN, Khattak FM, Jabbar MA. Effect of feeding flax and two types of antioxidants on egg production, egg quality, and lipid composition of eggs. J Appl Poult Res. 2009;18:541–51.View ArticleGoogle Scholar
- Selvaraj RK, Cherian G. Changes in delayed type hypersensitivity, egg antibody content and immune cell fatty acid composition of layer birds fed conjugated linoleic acid, n-6 or n-3 fatty acids. Can J Anim Sci. 2004;84:221–8.View ArticleGoogle Scholar
- Ulmer-Franco AM, Cherian G, Quezada N, Fasenko GM, McMullen LM. Hatching egg and newly hatched chick yolk sac total IgY content at three broiler breeder flock ages. Poult Sci. 2012;91:758–64.View ArticlePubMedGoogle Scholar
- SAS Institute. SAS User’s Guide. Statistics, Release 9.2. Cary: SAS Inst. Inc; 2001.Google Scholar
- Steel RGD, Torrie JH. Principles and Procedures of Statistics: A Biometrical Approach. 2nd ed. Toronto: McGraw-Hill Book Co; 1980.Google Scholar
- Valkonen E, Venäläinen E, Tupasela T, Hiidenhovi J, Valaja J. Effect of camelina sativa meal on fatty acids composition of egg yolk and sensory quality of eggs. World Poult Sci J. 2006;62:146–56.Google Scholar
- Rokka T, Alen K, Valaja J, Ryhanen EL. The effect of Camelina sativa enriched diet on the composition and sensory quality of hen eggs. Food Res Intl. 2002;35:253–6.View ArticleGoogle Scholar
- Cherian G. Modifying Egg Lipids for Enhancing Human Health. In: Cherian G, Poureslami, editors. Fats and Fatty Acids in Poultry Nutrition and Health, vol. 4. Leicestershire: CONTEXT Products Ltd; 2012. p. 57–68.Google Scholar
- Virtanen JK, Mursu J, Tuomainen T-K, Virtanen HEK, Voutilainen S. Egg consumption and risk of incident type 2 diabetes in men: the Kuopio Ischaemic Heart Disease Risk Factor Study. Am J Clin Nutr. 2015;101:1088–96.View ArticlePubMedGoogle Scholar
- Betti M, Perez TI, Zuidhof MJ, Renema RA. Omega-3-enriched broiler meat: 3. Fatty acid distribution between triacylglycerol and phospholipid classes. Poult Sci. 2009;88:1740–54.View ArticlePubMedGoogle Scholar
- Cherian G. Eggs and health: nutrient sources and supplement carriers. In: Watson RR, editor. Complementary and Alternative Therapies and the Aging Population. Cambridge: Academic; 2009. p. 333–46.View ArticleGoogle Scholar
- Kris-Etherton PM, Taylor DS, Yu-Poth S, Huth P, Moriarty K, Fishell V, Hargrove RL. The polyunsaturated fatty acids in the food chain in the United States. Am J Clin Nutr. 2000;71:179S–88.
- Hall JA, Jha S, Cherian G. Dietary n-3 fatty acids decrease the leukotriene B4 response ex vivo and bovine serum albumin-induced footpad swelling index in New Hampshire hens. Can J Anim Sci. 2007;87:373–80.View ArticleGoogle Scholar
- Cherian G, Bautista-Ortega J, Goeger DE. Maternal dietary n-3 fatty acids alter cardiac ventricle fatty acid composition, prostaglandin and thromboxane production in growing chicks. Prostaglandins Leukot Essent Fatty Acids. 2009;80:297–303.View ArticlePubMedGoogle Scholar
- Bullock C. Maternal Diet and Essential Fatty Acid Metabolism in Progeny Chickens, M.S Thesis. Corvallis: Oregon State University; 2013.Google Scholar