Effects of Sea Buckthorn Fruits on Production Performance and Egg Quality of Chicken Gallus gallus domesticus ‘Wuhei’ABSTRACT
This study was conducted to examine the effects of sea buckthorn fruits (SBF) on the production performance and egg quality of chickens during the initial stage of the laying period. A total of 160 chickens were randomly divided into four groups, with 40 birds in each group. The chickens of the control group were fed with a standard feed, while those in the three other groups were fed with the feed supplemented with 1, 2 and 4 g/kg of SBF powder, respectively. It was found that the dietary intake of SBF powder had little effect on the initiation time of egg production, laying rate, average egg weight, yolk weight, yolk ratio, eggshell thickness, albumen texture and yolk texture of the eggs; but the addition of SBF powder at dosages of 2 and 4 g/kg caused significant increase in yolk color and HU values (p<0.01), and the 4 g/kg dose resulted in a marked increase in albumen height (p<0.01). Importantly, dietary intake of SBF powder reduced the cholesterol level in the eggs (p<0.01). Moreover, supplementation of SBF powder into the feed increased the contents of saturated fatty acids (SFAs), monounsaturated fatty acids (MUFAs), and polyunsaturated fatty acids (PUFAs), with the PUFAs/SFAs ratio enhancing (p<0.001) by adding 2 g/kg SBF powder to the feed. In summary, our study suggests that SBF has potential for application in the production of high-quality eggs with reduced cholesterol, enhanced PUFAs/SFAs ratio, and better aroma and taste.
Keywords:
Sea buckthorn; laying chickens; production performance; egg quality; cholesterol; fatty acid
INTRODUCTION
Eggs are a cheap, widely available, affordable, and consumed food worldwide. They are a rich source of proteins and amino acids (including all the 8 essential amino acids), and essential nutrients such as unsaturated fatty acids, carotenoids and riboflavin, all of which are beneficial to human health (Zeisel & Corbin, 2012; Rehault-Godbert et al., 2019; Myers & Ruxton, 2023). For example, eggs have a relatively higher amount of unsaturated (monounsaturated plus polyunsaturated) fatty acids (5.31 g per 100 g of whole egg), including omega-3 fatty acids (n-3 PUFA), arachidonic acid (AA) and docosahexaenoic acid (DHA) (Rehault-Godbert et al., 2019). The dietary intake of n-3 PUFA reduces the risk of heart disease, inhibits the growth of prostate and breast cancer, and is required for normal fetal brain and visual development (Derbyshire, 2018; Lewis et al., 2000). Long-chain polyunsaturated fatty acids can also decrease allergic sensitization of infants (Miles et al., 2021). However, it is notable that despite many nutritional benefits, eggs contain high levels of dietary cholesterol (Wang et al., 2019), which has been repeatedly related with a higher risk to cardiovascular disease and diabetes (Shi et al., 2011; Shin et al., 2017; Chen et al., 2021; Wang et al., 2021; Yakti et al., 2024). Hence, improvement of the nutritional elements in eggs has increasingly become an important research topic (Park et al., 2015; Grcevic et al., 2019; Omri et al., 2019).
Plants and plant-derived compounds have been reported as effective feed additives to improve the production of livestock and poultry (Dannenberger et al., 2018; Qin et al., 2020; Fievez et al., 2021; Bayir & Yanik, 2025). Dietary supplementation of a phytogenic feed additive containing a blend of poplar (Populus deltoides) and eucalyptus (Eucalyptus citriodora) leaves was shown to enhance both cell-mediated and humoral immune responses, and increase milk production in buffaloes (Dey et al., 2021); mulberry leaf flavonoids were found to promote egg production and antioxidant capacity in chickens (Huang et al., 2021); and phytogenic tannins were demonstrated to improve rumen metabolism and ruminant nutrition (Patra & Saxena, 2011). Sea buckthorn (Hippophae rhamnoides L.), a deciduous shrub that can grow under harsh conditions such as frost and drought, contains many bioactive compounds, including polyphenols, carotenoids, flavonoids, and unsaturated fatty acids. It has been widely used in feed additives for cattle (Fievez et al., 2021), pigs (Dannenberger et al., 2018), ducks (Yao et al., 2023) and chickens (Chand et al., 2018; BenMahmoud et al., 2021). The fruits, seeds and leaves of sea buckthorn have been reported to increase egg weight and laying rates in poultry (Biswas et al., 2010; Chand et al., 2018; Yao et al., 2023). In addition, supplementation of sea buckthorn pomace to the feed of laying chickens has been found to improve the lipid oxidation status of egg yolks, which eventually leads to the extension of the shelf life of eggs (Panaite et al., 2021). However, their potential as feed additives in improving nutritional elements in eggs remains largely unexplored. The aim of this study is thus to examine the effects of the supplementation of sea buckthorn fruit powder on the production performance, and egg quality of the five black chicken (Gallus gallus domesticus ‘Wuhei’), a southern China specialty breed.
MATERIALS AND METHODS
G. gallus domesticus ‘Wuhei’ has a breeding history of over 1,300 years in China. Because of its distinctive “five black” characteristics, i.e. black feathers, black skin, black meat, black bones, and black internal organs, it stands as a unique and globally rare poultry breed. The eggs it lays have green shells, making it an exceptional avian treasure.
Experimental design and chicken husbandry
The study was approved by the Ethics Committee of Experimental Animals of Kashi University (approved number: 2025016). A total of 160 chickens of 14 weeks of age were purchased from Guizhou Longxiang Breeding Co., Ltd (China). The chickens were acclimated for a week, and then divided at random into four groups, each consisting of 40 chickens. All the chickens were raised in open sided henhouses under similar environmental conditions, with a cycle of 16 h light/8 h darkness at Yaoshang Village in Guiyang City, Guizhou Province. The poultry houses have a layer of dried straw covering the ground, and food and water containers hanging in the air to ensure the hygiene of feed and water. Chickens of group one (control) were fed with the standard feed for laying hens based on corn, wheat, and soybean (Table 1), while chickens of other three groups were fed the same feed, supplemented with 1 (T1), 2 (T2) or 4 (T3) g/kg of dried sea buckthorn fruit (SBF) powder, respectively (Fig. 1).
A schematic diagram of the experiment. The time points for feeding and sampling are shown. Administration of the feed supplemented with sea buckthorn fruit powder began on the 15th week, and eggs were sampled from groups 1, 2, 3 and 4 on the 24th, 25th, 26th and 27th weeks, respectively.
SBF powder was purchased from Anhui Changfuguo Pharmaceutical Co., Ltd (Bozhou, China). Feed and water were provided ad libitum. All chickens were fed twice a day (usually at 9:00 am and 17:00 pm).
Production performance assay
The experiment lasted for 12 weeks, from September 2024 to December 2024. The body weights (BW) of chickens were determined at the beginning (15 weeks of age) and at the end (27 weeks of age) of the experiment. The health and laying of eggs were checked every day till the end of the experiment. The eggs laid were collected every morning before feeding, numbered and weighed. The average daily feed intake [ADFI (g/d) = cumulative feed intake/(number of birds × number of days)], average egg weight (AEW = total daily egg mass/laying number), laying rate [LR (%) = (laying number/layer number) × 100], and feed conversion ratio [FCR = total feed intake/total egg weight] of the laying chickens were calculated, and used to monitor their production performance.
Egg quality assay
From week 24 and for 4 weeks, when the laying rate was over 1/3 for every group, to the end of experiments, 20 eggs from each group were sampled randomly every week. Ten out of the 20 eggs were used for egg quality assays, and 10 were used for sensory evaluation. All the analyses were completed within 24 h of egg collection. The eggs were weighed and broken on Petri plates to determine albumen height. The measurements of albumen height and egg weight were used to calculate the Haugh unit score (HU). Yolk color intensity was evaluated and scored according to a yolk color fan (DSM, Heerlen, Netherlands) (1, light yellow; 15, orange). The thickness of eggshell was measured at three different locations (middle, broad and narrow ends) with the use of a digital micrometer gauge and expressed as the mean value.
Sensory Evaluation
The 10 eggs of each group were boiled at 100 ºC for 15 min for sensory evaluation. The boiled eggs were cooled under running cold water for 15 min and then evaluated on a 5-point hedonic scale: 5, very high quality; 4, high quality; 3, satisfactory quality; 2, unsatisfactory quality; and 1, poor quality. The following sensory quality attributes were evaluated: appearance (including the appearance of the whole egg and its longitudinal cross-section), aroma, albumen texture, yolk texture, and taste. The analysis was conducted by 5 trained panelists selected for their sensory sensitivity. The panelists did not know which group the eggs came from and assessed the hard-boiled eggs under the same environments.
Chemical analysis
A total of 10 fresh eggs were randomly sampled from each group on week 27, with 5 eggs being used for the determination of cholesterol, and 5 eggs the measurement of fatty acids.
The cholesterol content in the eggs was determined according to GB5009.128-2016 (Chinese Standard GB 5009.128-2016, 2016), the national standard for food safety. In short, all 5 eggs were broken and mixed thoroughly. An aliquot of 10 g of the sample was mixed with 30 mL of 100% ethanol and 10 mL of 60% KOH, and saponified at 100 ºC for 1 h. After saponification, the fat was extracted with petroleum ether-anhydrous ethanol mixture, and subjected to analysis using gas chromatograph (TRACE1300, Thermo Scientific, USA). The column used was TG-5MS (30 m × 0.25 mm × 0.2µm). The injector temperature was set at 280 ºC, and the detector temperature was set at 300ºC. The initial temperature program was 20ºC for 0.5 min, and then the temperature was increased at a rate of 40 ºC/min until 280ºC, which lasted for 10 min. The carrier gas was nitrogen, the flow rate was 1.0 mL/min, and the injection volume was 1.0 µL. The analysis was repeated three times.
The fatty acids in the eggs were analyzed according to GB5009.168-2016 (Chinese Standard GB 5009.168-2016, 2016). In brief, 5 eggs were broken and mixed thoroughly. An aliquot of 10 g of the sample was taken into a 250 mL beaker, mixed with 100 mg of pyrogallic acid and 2 mL of 95% ethanol, and shaken thoroughly. The fat was extracted with petroleum ether-anhydrous ether mixture, and subjected to analysis using a gas chromatograph (TRACE1300, Thermo Scientific, USA). The column used was SP-2560 (100 m × 0.25 mm × 0.2µm). The injector temperature was set at 230ºC, and the detector temperature was set at 250 ºC. The initial temperature program was 100 ºC for 13 min, and then the temperature was increased at a rate of 10 ºC/min to 180 ºC, which lasted for 6 min, followed by 180ºC~200ºC, 1ºC/min, 20 min; and 200ºC~230ºC, 4ºC/min,10.5 min. The carrier gas was nitrogen, the flow rate was 1.0 mL/min, and the injection volume was 1.0 µL. The analysis was repeated three times.
Statistical analysis
All data were analyzed using the general linear model program of SPSS 27. For data that met the assumptions of variance analysis, one-way ANOVA with LSD (Least Significant Difference) was applied, and Duncan’s multiple comparison test was used for further analysis. For data that did not meet the assumptions of variance analysis, the Kruskal-Wallis test was conducted, and then followed by the Mann-Whitney test. The results of each group were expressed as the mean, and the standard error of the mean (SEM). A p-value of <0.05 was considered statistically significant.
RESULTS
All the chickens were maintained in good health throughout the experimental period. The chickens of all 4 groups were found to start laying eggs at the age of 18th week. Fig. 2 shows the laying rate of the chickens at different weeks of the experiment in all the groups, and Table 2 presents the effects of different levels of SBF powder on feed intake and production performance. The laying rate in Table 2 was the average laying rate from the beginning of egg-laying to the end of the trial.
Curves showing the weekly laying rate of the chickens at different weeks of the experiment. The chickens of all groups started laying eggs at the 18th week of age, and no significant changes in laying rate were observed.
It was found that the supplementation of SBF powder had little influence on the average daily feed intake, feed conversion ratio, initiation time of egg production, and laying rate of the chickens, but the supplementation of SBF powder at a dosage of 4 g/kg caused a decrease in the final body weight (p<0.05).
Table 3 shows the effects of supplementation of SBF powder on egg quality. No significant difference was observed on the average egg weight, eggshell thickness, yolk weight, yolk ratio and egg shape index of the chickens. By contrast, the addition of SBF powder at dosages of 2 g/kg and 4 g/kg resulted in a significant increase in the yolk pigmentation and HU values (p<0.01), and addition of SBF powder at a dosage of 4 g/kg led to a marked increase in the albumen height (p<0.01).
The effects of different levels of SBF powder on cholesterol and fatty acids in the eggs are shown in Table 4. It was found that the addition of SBF powder into the feed even at a dosage of 1 g/kg caused a significant reduction of egg cholesterol (p<0.01), with greater reductions in groups receiving higher doses. In contrast, the addition of SBF powder at dosages of 2 g/kg and 4 g/kg resulted in a significant increase in saturated fatty acids (SFAs), monounsaturated fatty acids (MUFAs) and polyunsaturated fatty acids (PUFAs; p<0.01). It was notable that the addition of SBF powder into the feed at a dosage of 2 g/kg caused a marked increase in the PUFA/SFA ratio.
The effects of SBF powder on average intensity values of egg sensory attributes are presented in Table 5. The dietary SBF powder showed no negative effects on the general appearance, albumen texture and yolk texture of the eggs (p>0.05), but it did improve the aroma and taste of the eggs (p<0.05).
DISCUSSION
The pursuit of healthier food sources has become a common trend among consumers. Plants and their compounds are increasingly being used to improve the production of livestock and poultry (Dannenberger et al., 2018; Qin et al., 2020; Fievez et al., 2021; Bayir & Yanik, 2025). This study examined the effects of dried SBF powder supplemented into feed on the production performance and egg quality of Wuhei chickens during the initial stage of the laying period. It has been reported by Chand et al. (2018) that supplementation of sea buckthorn seed into the feed causes a marked decrease in yolk cholesterol among Rhode Island Red×Fayoumi laying hens. This study clearly shows that dietary intake of SBF powder is capable of significantly reducing the cholesterol level in the eggs, agreeing with the results of Chand et al. (2018). In addition, we also show that the addition of SBF powder into the feed increases SFA, MUFA and PUFA contents. Notably, the PUFAs/SFAs ratio was apparently enhanced by adding 2 g/kg SBF powder to the feed. Reduced cholesterol and enhanced PUFA/SFA ratio are desirable egg qualities, as both cholesterol and SFA have the potential to increase cholesterolemia (Vlaicu et al., 2021).
Cholesterol in eggs is either absorbed from feed or synthesized by chickens themselves. Sea buckthorn is a rich source of phytosterols, mainly β-sitosterol (Cenkowski et al., 2006). It has been reported that phytosterols have anticholesterolemic activity (Khan et al., 2012) by blocking the absorption of cholesterol in the intestine of chickens (Davis et al., 1955). Besides, phytosterols have lower hydro-solubility than cholesterol, thus the bioavailability of β-phytosterols for intestinal micelles is greater than that of cholesterol, which may eventually lead to a reduced cholesterol absorption rate (Shahid et al., 2015). Moreover, phytosterols can also reduce hepatic biosynthesis of cholesterol, thus limiting its amount in eggs (Golimowski et al., 2022). In contrast to cholesterol absorption in vitro and production in vivo, PUFAs are not synthesized by chickens themselves, and must be supplied in the diets. It is known that sea buckthorn is among the richest sources of α-linolenic acid, linoleic acid and palmitoleic acid, which may contribute to the enrichment of PUFAs in the eggs (Favé et al., 2004).
It has been reported that sea buckthorn can improve egg weight and laying rate in poultry across multiple experimental settings (Biswas et al., 2010; Chand et al., 2018; Vlaicu et al., 2021). Our study shows that SBF powder has little effects on the initiation time of egg production, laying rate, average egg weight, eggshell thickness, yolk weight, yolk ratio and egg shape index. The discrepancy may be due to the species and/or different ages of the chickens used. Notably, our study demonstrates that the addition of SBF powder to the feed improves the yolk pigmentation, HU values, and albumen height of the chicken eggs. Improvement in egg yolk color has been attributed to the transfer of carotenoids, xanthophylls, and chlorophylls in diets to the yolks (Öztürk et al., 2024). Thus, it is possible that the increase in yolk pigmentation caused by feeding SBF powder may also be due to the higher content of carotenoids in sea buckthorn fruits (Ren et al., 2020). Many people prefer eggs with intense yolk color, as it is often linked to perceptions of nutritional value and freshness of eggs (Hisasaga et al., 2022). HU value and albumen height are both indicators of egg freshness. It has been found that supplementation of dried rosehip fruit pomace to the diets of laying hens can promote the antioxidant potential of eggs, thus improving eggs stability and protecting the oxidation of egg fats (Mierlita et al., 2024). Similarly, the addition of sea buckthorn pomace to the feed of laying hens can improve the lipid oxidation status of egg yolks, thereby significantly reducing the peroxide value. A decrease in the peroxide value can enhance the retention rate of nutrients in eggs and slow down the accumulation of harmful products, ultimately extending the shelf life of eggs (Panaite et al., 2021). Thus, we believe that SBF powder can also improve the antioxidant potential of eggs, thereby helping to maintain them fresh.
Previous studies have demonstrated that supplementation of rapeseed products to layer diets can compromise the sensory attributes of eggs, in particular by imparting a fishy taint to eggs (Elangovan et al., 2001; Goldberg et al., 2016). In the present study, the addition of SBF powder into feeds apparently improved the aroma and taste of the eggs, while it had little effect on the general appearance, albumen texture, and yolk texture. This seems to be the first report that sea buckthorn fruit can optimize sensory attributes of eggs such as aroma and taste.
Finally, we show that the addition of SBF powder into feeds has no effect on the average daily feed intake, indicating that SBF powder does not adversely affect the appetite or feed consumption of the laying chickens. However, the feed with 4 g/kg SBF powder results in reduced final body weight of the chickens, suggesting that a higher dosage of SBF powder may not be conducive to chicken growth. The reason for this is unknown at present. One possibility is that a higher dosage of SBF powder may impair metabolism of the laying chickens, which will in turn affect their growth. This issue deserves further study.
CONCLUSION
The present study explored the effects of sea buckthorn fruits on the production performance and egg quality of five-black chickens, a specialty breed from China. It shows that sea buckthorn fruits as feed additives can improve egg quality by reducing cholesterol levels and increasing PUFAs/SFAs ratio during the initial stage of the laying period. The addition of sea buckthorn fruits to feed can also improve the aroma and taste of eggs.
ACKNOWLEDGEMENTS
This work was supported by grants (022023184, 02202350) from the research start-up funds of Kashi University to SZ.
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FUNDING
This work was supported by grants (022023184, 02202350) from the research start-up funds of Kashi University to SZ.
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DATA AVAILABILITY STATEMENT
The data supporting the findings of this study are available from the corresponding author upon reasonable request.
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DISCLAIMER/PUBLISHER’S NOTE
The published papers’ statements, opinions, and data are those of the individual author(s) and contributor(s). The editor(s) disclaim responsibility for any injury to people or property resulting from any ideas, methods, instructions, or products referred to in the content.
The data supporting the findings of this study are available from the corresponding author upon reasonable request.
