Flaxseed Oil for Omega 3 Fatty Acids Enrichment in Eggs and Performance in Laying Chickens

B. Prakash1,*, S.V. Rama Rao1, M.V.L.N. Raju1, S.S. Paul1, A. Kannan1, Y. Praduman2, P.K. Santosh1
1ICAR-Directorate of Poultry Research, Rajendranagar, Hyderabad-500 030, Telangana, India.
2ICAR-Indian Institute of Oilseeds Research, Rajendranagar, Hyderabad-500 030, Telangana, India.
Background: The beneficial effects of omega-3 fatty acids on growth, health and immune function for human is well established. Supplementation of various n-3 fatty acids into the diet of laying hens is a nutritional attempt to increase the levels of n-3 PUFA in the eggs. Therefore, the present study was carried out to determine the effect of dietary supplementation of flaxseed oil (SFO) as a source of omega-3 polyunsaturated fatty acids, on the fatty acid composition of egg, laying performance and egg stability upon storage in laying hens. 

Methods: Laying hens of 30-week age (n=168) were selected and randomly distributed into two dietary groups consisting of 14 replicates each having 6 birds in each replicate. Iso-nitrogenous and iso-caloric two diets were prepared with and without FSO. Daily recording of eggs, feed intake and egg collection was carried out throughout the experiment. The eggs were subjected for fat extraction and estimation of fatty acid composition. 

Result: The higher (P<0.05) feed intake and lower (P<0.05) egg production in groups fed FSO supplemented diet compared to those groups fed control diet. Further, improved (P<0.05) feed conversion ratio among the groups fed control diet compared to those groups fed FSO supplemented diet. Eggs contained higher (P<0.05) á-linolenic acid, eicosenoic acid, n3 fatty acids in groups fed FSO supplemented diets compared control diet. The stability parameters did not differ by supplementing FSO compared to groups fed control diet. Therefore, it is concluded that FSO can be supplemented up to 6% for feeding of laying hens for producing higher omega 3 fatty acids in eggs. Further, supplementation of 6% FSO did not affect the egg quality parameters upon storing for 21 days at 4°C. Background: The beneficial effects of omega-3 fatty acids on growth, health and immune function for human is well established. Supplementation of various n-3 fatty acids into the diet of laying hens is a nutritional attempt to increase the levels of n-3 PUFA in the eggs. Therefore, the present study was carried out to determine the effect of dietary supplementation of flaxseed oil (SFO) as a source of omega-3 polyunsaturated fatty acids, on the fatty acid composition of egg, laying performance and egg stability upon storage in laying hens. 
The beneficial effects of omega-3 (n-3) polyunsaturated fatty acids (PUFA) on growth, health and immune function for humans is well established (Goyal et al., 2014; Lee et al., 2019). Therefore, supplementation of various n-3 fatty acids into the diet of laying hens has been a nutritional attempt to increase the levels of n-3 PUFA in the chicken eggs (Oliveira et al., 2010) and meat (Panda et al., 2015). Dietary supplements for n-3 PUFA are fish oil, flaxseed, or microalgae (Alagawany et al., 2019) have been extensively studied. Though fish oil is rich in n-3 PUFA, but there are negative reports that laying hens fed with fish oil-added diet produced fishy odor eggs and contains heavy metals in eggs (Coorey et al., 2015). The dietary microalgae have been recently marketed as a source of n-3 PUFA, but the relatively higher production cost and uneconomical (Fraeye et al., 2012). The dietary flaxseed oil has been used to produce n-3 enriched eggs or meats in the poultry industry (Oliveira et al., 2010; Petrovic et al., 2012). Nevertheless, most of the studies shows that the supplementation of SFO is lower, even less than 1% in the diet (Lee et al., 2021). However, it has been reported that the supplementation of FSO linearly increases the omega 3 fatty acids in eggs. Further, as PUFA in layer diets increases the susceptibility of eggs to lipid oxidation (Cherian et al., 2007) increases. Therefore, the present study was conducted to determine the performance, egg stability and fatty acid content of eggs in laying hens fed higher quantity FSO.
White Leghorn layers (F-line) 30-week age (n=168) were housed and randomly distributed into two dietary groups consisting of 14 replicates each having 6 birds in each replicate. The experiment was carried out from 33 to 40 weeks of age at ICAR-Directorate of Poultry Research, Hyderabad from August 2021 to February 2022. Iso-nitrogenous and iso-caloric two diets were prepared with and without FSO (6%; Table 1). Daily recording of eggs, feed intake and egg collection was carried out throughout the experiment. The crude protein levels in both the diets were analysed by the Kjeldahl method after acid hydrolysis using AOAC (2005).
 

Table 1: Ingredient and nutrient content of experimental diets.


       
The eggs were subjected extraction of fat and estimation of fatty acid composition using Gas chromatograph. Briefly, the CP-Sil 88 capillary column (50 m x 0.32 mm x 0.2 µm) was used at oven temperature (60°C, 1 min, 25°C/min to 160°C, 28 min, 25°C/min to 190°C, 17 min, 25°C/min to 220°C, 10 min) using flame ionization detector and detector temperature was 250°C (Araujo et al., 2010). The FSO and oil extracted from egg yolk was subjected for estimation of fatty acid composition. Further, the eggs were stored during the experiment at weekly interval (14 eggs from each treatment, each from each of the replicate) for estimation of egg quality parameters of eggs (stored for 1, 2 and 3 weeks) during the experiment for determining the egg weight, density, Hough unit, shell weight and shell thickness.
               
The variations in data of different parameters were analyzed using one-way analysis of variance procedure of SAS version 9.2 (2008; SAS Institute Inc., Cary, North Carolina, USA). The model included the different treatments were designated as the fixed factors and response  variables were taken as the dependent variables.
The FSO was subjected for estimation of fatty acid composition and was fond to have contain α-linolenic acid (C18:3n-3) (52.52%), Linoleic acid (C18:2n-6) (14.27%), Oleic acid (C18:1n-9) (21.13%), Stearic acid (C18:0) (5.58%) and Palmitic acid (C16:0) (6.15%) of total fatty acids in FSO. The values of fatty acids recorded in FSO in the present study are in agreement with the reported values of Lee et al., (2021).
       
The feed intake was higher (P<0.05) in FSO supplemented groups compared to those groups fed control diet (Table 2 and 3) during both the periods (33-36 and 37 to 40 weeks). Whereas, significantly lower hen day egg production (P<0.05) was recorded in FSO supplemented groups compared to those groups fed control diet. Improved (P<0.05) feed conversion (FCR) ratio among the groups fed control diet compared to those groups fed FSO supplemented diet. It is well established that the fat is used to decrease dustiness and enhance energy density in poultry diets. However, supplementation of flaxseed oil at 4 per cent improved the body weight gain and feed efficiency of broiler chickens with no adverse effects on carcass parameters (Bharat et al., 2017). The increased feed intake and reduced hen day egg production in laying hens at higher levels of FSO (4.5%) supplementation is also reported by Promila et al., (2017). However, the body weight gain, feed consumption as well as feed conversion ratio are not affected by dietary incorporation of FSO up to 2 and 3% in broiler chickens (Panda et al., 2015).
 

Table 2: Effect of feeding diet containing flaxseed oil on feed intake and egg production in IWF line during 33-36 weeks.


 

Table 3: Effect of feeding diet containing flaxseed oil on feed intake and egg production in IWF line during-40 weeks.


       
Egg yolk contained significantly (P<0.05) higher α-linolenic acid, eicosenoic acid, n3 fatty acids in groups fed FSO supplemented groups. The ratio of n6 to n3 decreased (P<0.05) significantly in groups fed FSO supplemented diets compared to those groups fed control diet (Table 4). It is established that the dietary supplementation of FSO linearly increases the percentages of heptadecanoic acid, eicosatrienoic acid and DHA in egg yolks (Seyyed and Hasan, 2018; Lee et al., 2021). Similar to our findings, the ratio of n-6 to n-3 fatty acids in egg yolks decreases with increase the supplemental levels of FSO in the diets (Lee et al., 2021). The main biological role of α-linolenic acid is to serve as a substrate for the synthesis of EPA and DHA (Liang et al., 2017). Laying hens have the ability to elongate and desaturate α-linolenic acid to the functional EPA and DHA (Ehr et al., 2017).  Therefore, it is evident from the present study that the feeding diets supplemented FSO, which is rich in á-linolenic acid resulted in higher content of EPA and DHA.
 

Table 4: Effect of flaxseed oil in the diet of layer birds on egg fatty acid composition.


               
In present experiment, none of the egg stability parameters were differed by supplementing FSO compared to those group fed control diet up to 21 days of storage in the refrigerator at 4°C (Table 5). PUFA rich eggs are more susceptible to oxidation as PUFAs have several double bonds (Wang et al., 2017). It has been reported that the lipid oxidation is a process that affects egg yolk lipid stability during storage (Faitarone et al., 2016; Omri et al., 2019) and affects egg quality parameters (Omri et al., 2019). However, the egg quality parameters did not vary and remained comparable with those groups fed control diet. 
 

Table 5: Effect of flaxseed oil on egg quality parameters that were stored up to 21 days in refrigerator at 4°C.

Flax seed oil can be supplemented up to 6% for feeding of laying hens for producing higher omega 3 fatty acids in eggs. Further, supplementation of 6% flax seed oil did not affect the egg quality parameters upon storing for 21 days at 4°C. Supplementation of 6% flax seed oil did not affect the egg production. However, the higher feed intake and poor feed efficiency was recorded in laying hens fed flax seed oil supplemented diet.
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