Enhancing the internal quality of lohmann brown eggs through carrot leaf extract supplementation

*Corresponding author: Ni Wayan Siti Copyright © 2025 Author(s) retain the copyright of this article. This article is published under the terms of the Creative Commons Attribution License 4.0. Enhancing the internal quality of lohmann brown eggs through carrot leaf extract supplementation Dinda 1, I Gusti Nyoman Gde Bidura 2, Ni Wayan Siti 2, * and Ni Wayan Ayu Ningsih 2 1 Master’s Program, Faculty of Animal Husbandry, Udayana University, Badung, Bali Indonesia. 2 Faculty of Animal Husbandry, Udayana University, Badung, Bali, Indonesia. World Journal of Biology Pharmacy and Health Sciences, 2025, 24(02), 533-538 Publication history: Received 08 October 2025; revised on 22 November 2025; accepted on 24 November 2025 Article DOI: https://doi.org/10.30574/wjbphs.2025.24.2.1028 Abstract This study aims to examine the supplementation of carrot leaf water extract through drinking water to improve the internal quality of lohmann brown chicken eggs. The experimental design used was a completely randomized design (CRD) consisting of four treatments and five replicates, with each replicate using four lohmann brown chickens. The treatments given were lohmann brown chickens given drinking water without carrot leaf water extract (A), lohmann brown chickens given drinking water with 2% carrot leaf water extract (B), lohmann brown chickens given drinking water with 4% carrot leaf water extract (C), and lohmann brown chickens given drinking water with 6% carrot leaf water extract (D). The variables observed were egg weight, egg density, egg index, egg white index, egg yolk index, egg yolk color, Haugh unit, pH, cholesterol, and beta-carotene. The results of the study showed that the supplementation of carrot leaf extract had a significant effect (P<0.05) on egg white index, Haugh unit, egg yolk color, cholesterol, and betacarotene, while the other variables showed no significant difference (P>0.05). Based on the research results, it can be concluded that the supplementation of carrot leaf water extract can increase the egg white index, Haugh unit, egg yolk color, and beta-carotene content and reduce cholesterol levels in the egg yolk. Keywords: Lohmann Brown; Carrots; Egg Yolk Color; Cholesterol; Beta Carotene 1. Introduction Eggs are a source of animal protein from lohmann brown laying hens that are in high demand because they contain essential nutrients and are affordable. Currently, consumers are beginning to pay attention to the quality of the eggs they consume, with a tendency to prefer eggs with high internal quality, such as high Haugh units, bright yolk color, high β-carotene, and low cholesterol. Efforts to improve the internal quality of eggs require the provision of high-quality feed and feed additives. Antibiotic growth promoters (AGPs) are feed additives for poultry that have been banned because they can leave residues and are harmful to consumer health [1]. Therefore, natural feed supplements from natural ingredients such as carrot leaf waste are needed as a substitute for AGPs to increase the production and quality of free range chicken eggs. Carrot leaves (Daucus carota) contain compounds that are almost identical to those found in carrots, including phytochemicals such as flavonoids, phenols, terpenoids, steroids, tannins, polyphenols, carotenoids, and β-carotene, which can reduce fat, act as antioxidants, prevent cancer, and boost the immune system of laying hens [2]. According to [3], carrot production in Indonesia reached 720.09 thousand tons, an increase of 10.64% from 650.86 thousand tons in 2020. Research by [4] showed that water extract of carrot leaves obtained through cold maceration at a concentration of 2% had a strong inhibitory effect on Staphylococcus aureus bacteria and could improve yolk color and reduce World Journal of Biology Pharmacy and Health Sciences, 2025, 24(02), 533-538 534 cholesterol levels in the yolk [5]. Therefore, this study was conducted to examine the effect of carrot leaf water extract in improving the internal quality of Lohmann chicken eggs. 2. Materials and methods 2.1. Materials The chickens used in this study were 80 lohmann brown chickens aged 30 weeks with an average weight and standard deviation of 1952.13±146.55 g. The cages used were battery colony cages. Feed and drinking water were provided ad libitum and consisted of 44.38% yellow corn, 36.98% concentrate, 18.49% rice bran, and 0.15% premix. The concentrate used was CP124 layer concentrate produced by PT Charoen Pokphand Indonesia (CPI). Table 1 Nutritional content of feed ingredients for lohmann brown chickens Feed Composition % ME CP CF CFib Ca P Met Lys Corn 44.38 1495 3.81 1.82 0.98 0.01 0.13 0.08 0.09 CP124 Concentrate 36.98 924 11.46 0.74 3.33 4.44 0.19 0.30 0.63 Rice Bran 18.49 444 2.22 0.43 4.72 0.04 0.19 0.05 0.08 Premix 0.15 - - - - - - 0.05 0.05 Total (%) 100 2863 17.49 2.99 9.02 4.49 0.50 0.42 0.80 2.2. Carrot Leaf Extract Production Process Preparation of carrot leaf extract (CLE) was carried out by using 1 kilogram of fresh carrot leaves, which were crushed in heated water at a 1:1 ratio. The mixture was left to stand for 24 hours before being filtered to separate the coarse leaf particles from the liquid extract. The resulting carrot leaf extract was then ready to be supplementation to lohmann brown chickens according to the experimental treatments Table 2 Laboratory Analysis Results of the Phytochemical Compounds in Carrot Leaf Aqueous Extract Parameter Content IC50 (ppm) 2172.77 Fhenol (mg/100g) 28.57 Flavonoids (mg/100g) 35.6033 Tannins (mg/100g) 29.30 Beta-carotene (mg/100g) 908.75 Saponins Weakly positive (+) Terpenoids Strongly positive (+++) Source: Food Analysis Laboratory, Faculty of Agricultural Technology, Udayana University 2.3. Experimental Design This study used a completely randomized design (CRD) with 4 treatments and 5 replicates. Each experimental unit consisted of 4 lohmann brown chickens. The treatments used were: lohmann brown chickens given drinking water without carrot leaf extract (A), given drinking water with 2% carrot leaf extract (B), given drinking water with 4% carrot leaf extract (C), and given drinking water with 6% carrot leaf extract (D). 2.4. Observed Variables • Egg weight Egg weight was obtained by weighing the eggs using a digital scale with an accuracy of 0.001g. • Egg specific gravity World Journal of Biology Pharmacy and Health Sciences, 2025, 24(02), 533-538 535 Egg specific gravity was calculated as the ratio of egg weight to egg volume. Egg volume was measured by immersing the egg in a measuring cylinder filled with water and recording the increase in water volume. • Yolk and Albumen Indices The yolk and albumen indices were determined by measuring the height and diameter of the yolk and albumen using a caliper. The yolk index was calculated according to the formula described in SNI 01-39262006 [6]. The yolk index was calculated as follows: Yolk Index = 𝑌𝑜𝑙𝑘 𝐻𝑒𝑖𝑔ℎ𝑡 (𝑐𝑚) 𝑌𝑜𝑙𝑘 𝐷𝑖𝑎𝑚𝑒𝑡𝑒𝑟 (𝑐𝑚) Albumen Index = 𝐴𝑙𝑏𝑢𝑚𝑒𝑛 𝐻𝑒𝑖𝑔ℎ𝑡 (𝑐𝑚) 𝐴𝑙𝑏𝑢𝑚𝑒𝑛 𝐷𝑖𝑎𝑚𝑒𝑡𝑒𝑟 (𝑐𝑚) • Egg yolk color Egg yolk color was measured using the Roche Yolk Color Fan, which ranges from 1 to 15. • Haugh Unit Haugh Unit values were calculated using the following formula: HU= 100 Log (H+7,57 – 1,7 W0,37) Explanation: HU = Haugh Unit, H = Height of thick albumen, W = Egg weight • Acidity (pH) Egg pH was measured using a pH meter by homogenizing the egg yolk and albumen together. • Cholesterol Yolk cholesterol content was analyzed using the UV-Vis spectrophotometric method (AOAC, 2005). Fat was extracted using a chloroform:methanol mixture (2:1), followed by saponification with KOH to isolate cholesterol. The results were expressed as milligrams of cholesterol per gram of egg yolk. • Beta-Carotene According to [7], β-carotene content was determined by extracting 0.10–0.50 g of sample using a mixture of acetone and petroleum ether, followed by centrifugation until a clear supernatant was obtained. The supernatant was washed with distilled water, dried with Na₂SO₄, and diluted with petroleum ether to a final volume of 10 ml. Absorbance was measured at 450 nm using a spectrophotometer, and total carotene was calculated using the formula: Total Carotene (µg/100 g) = 𝑇𝑜𝑡𝑎𝑙 𝑉𝑜𝑙𝑢𝑚𝑒 𝑥 𝐴𝑏𝑠𝑜𝑟𝑏𝑎𝑛𝑐𝑒 𝑥 100 0.2 𝑥 𝑆𝑎𝑚𝑝𝑒𝑙 𝑊𝑒𝑖𝑔ℎ𝑡 2.5. Data Analysis The research data were analyzed using analysis of variance. If there were significant differences (P<0.05) between treatments, then Duncan's multiple range test was used [8]. 3. Results and Discussion Based on the results of the study, the supplementation of carrot leaf aqueous extract had no significant effect (P>0.05) on egg weight. The average egg weight obtained in this study ranged from 61.414 g to 64.322 g, which is close to the normal egg weight of lohmann brown hens, ranging from 62.0 to 64.6 g/egg [9]. The relatively similar egg weights among treatments indicate that supplementation up to 6% did not affect protein absorption from the feed. This is consistent with [10], who reported that egg weight is influenced by the amount of dietary protein absorbed by the bird, laying age, and uniformity of sexual maturity. The average egg specific gravity of lohmann brown hens receiving carrot leaf aqueous extract ranged from 1.041 to 1.088, showing no significant difference (P>0.05). The similarity in egg specific gravity values between treatments was attributed to the non-significant differences in egg weight (Table 3). Table 3 Effects of Carrot Leaf Aqueous Extract Supplementation on Internal Egg Quality Variable Treatment1) SEM2) A B C D Egg Weight (g) 61.414a3) 64.131a 64.322a 62.391a 1.130 World Journal of Biology Pharmacy and Health Sciences, 2025, 24(02), 533-538 536 Egg Specific Gravity (g/ml) 1.041a 1.058a 1.088a 1.054a 0.030 Egg Index (%) 78.819a 78.462a 79.504a 76.473a 0.911 Yolk Index (cm) 0.329a 0.286a 0.323a 0.305a 0.014 Albumen Index (cm) 0.037c 0.051bc 0.058ab 0.064a 0.004 Haugh unit 56.412b 65.356ab 73.914a 75.797a 4.881 pH 7.786a 7.536a 7.528a 7.746a 0.144 Notes : A = drinking water without carrot leaf aqueous extract; B = drinking water with 2% carrot leaf aqueous extract; C = drinking water with 4% carrot leaf aqueous extract; D = drinking water with 6% carrot leaf aqueous extract.; Standard error of the treatment means (SEM).; Different superscript letters within the same row indicate significant differences (P < 0.05). The analysis also showed that the supplementation of carrot leaf aqueous extract did not significantly affect (P>0.05) the egg index. The relatively similar egg index values among treatments were closely related to the similarity in egg weights (Table 3). This finding aligns with [11], who stated that egg index is influenced by egg weight; the heavier the egg, the higher the resulting egg index, and vice versa. The egg yolk index of eggs treated with carrot leaf extract showed no significant effect (P>0.05). The yolk index values ranged from 0.29 to 0.33, which is considered normal and close to the standard yolk index value of 0.33–0.50. The similarity in yolk index, including yolk height and diameter, is likely due to the absence of significant differences in egg weight among treatments (Table 5.1), which influences yolk size [11]. The addition of 4% and 6% carrot leaf water extract resulted in significantly different egg white index values (P<0.05) compared to the control treatment. These differences may be attributed to the presence of phytochemicals such as tannins, flavonoids, polyphenols, and other bioactive compounds that act as antioxidants, protecting proteins from oxidative damage and helping maintain albumen viscosity [12]. [13] Also reported that phytochemical compounds act as antibacterial agents that inhibit microbial growth in eggs, thereby preventing protein denaturation The results of this study indicate that the supplementation of carrot leaf aqueous extract had a significant effect (P<0.05) on haugh unit values. As the level of carrot leaf extract increased, the Haugh unit value also increased (Table 3). This significant increase was attributed to the presence of phytochemical compounds such as flavonoids, which act as antioxidants and antibacterial agents, protecting proteins from degradation and inhibiting bacterial growth, thereby preventing protein denaturation. The average HU values obtained in this study were higher than those reported by [14], who administered herbavit (turmeric, moringa leaves, and ginger) to laying hens and obtained HU values of 64.32– 66.76. Table 4 Effect of Carrot Leaf Aqueous Extract Supplementation on Egg Yolk Color, Cholesterol, and Beta-Carotene Content of Lohmann brown Eggs Variable Treatment1) SEM2) A B C D Egg Yolk Color 7.6b3) 8.6a 8.8a 8.8a 0.224 Cholesterol (mg/g) 19.39a 18.27b 18,28b 18.14b 0.305 Beta-Carotene (mg/100 g) 0.037b 0.042a 0.041a 0.040a 0.001 Notes : A = drinking water without carrot leaf aqueous extract; B = drinking water with 2% carrot leaf aqueous extract; C = drinking water with 4% carrot leaf aqueous extract; D = drinking water with 6% carrot leaf aqueous extract.; Standard error of the treatment means (SEM).; Different superscript letters within the same row indicate significant differences (P < 0.05). The supplementation of carrot leaf aqueous extract did not have a significant effect (P>0.05) on egg pH. The phytochemical components such as flavonoids, polyphenols, and other bioactive compounds present in carrot leaf extract did not directly influence the buffering system responsible for maintaining pH stability during egg formation [12]. Statistical analysis of the yolk color of lohmann brown chickens fed 2%–6% carrot leaf extract showed significant differences (P<0.05) compared to A. This shows that yolk color tends to increase in line with increasing levels of carrot World Journal of Biology Pharmacy and Health Sciences, 2025, 24(02), 533-538 537 leaf extract. [15] stated that the carotenoid pigments from carrot leaves are absorbed into the blood, circulate throughout the body, and accumulate in the tissues, thereby affecting the yellow color of the eggs. The cholesterol content of eggs fed carrot leaf extract decreased significantly and was statistically different (P<0.05). The significant decrease in cholesterol was due to the flavonoid content in carrot leaves. Flavonoids can lower cholesterol levels by inhibiting cholesterol absorption in the intestines and binding bile acid formation reactions to inhibit fat absorption and increase fat excretion, including cholesterol, through feces [16]. The beta carotene content in egg yolks produced in this study showed significantly different results (P<0.05). The supplementation of carrot leaf water extract, which is high in beta-carotene, affected the amount of beta-carotene in egg yolks. [17] reported that beta-carotene from carrots is absorbed into the blood, then distributed throughout the livestock's body and accumulated in the tissues, thereby increasing the beta-carotene content in eggs. The increased beta-carotene content resulted in a high egg yolk color score and reduced cholesterol (Table 4). In line with the research by [15], the supplementation of carrot leaf supplements to lohmann brown chickens had a significant effect on egg yolk color, beta-carotene, and cholesterol 4. Conclusion Based on the results of the study, it can be concluded that the supplementation of carrot leaf water extract can increase the egg white index, haugh unit, yolk color, and beta carotene content, as well as reduce cholesterol levels in the egg yolk. Compliance with ethical standards Acknowledgements The author would like to thank the Dean of the Faculty of Animal Husbandry, Udayana University, for his assistance and support during the research. Disclosure of Conflicts of Interest We declare that there is no conflict of interest with any financial organization related to the material discussed in this paper. Statement of ethical approval The animal ethics committee approved the use of 80 lohmann brown hens aged 30 weeks for this study, from the Faculty of Veterinary Medicine, Udayana University, Badung, Bali, Indonesia References [1] Shaffitri, L. R., R. P. Perdana, N. Ilham, and E. A. Suryana. 2024. Implementation of feed business policies to support the development of the poultry industry. Agricultural Policy Analysis. 22(1): 1-15. [2] Dewi, A. C., M. M. D. Utami, T. Budiati, W. Suryaningsih, and N. Ningsih. 2024. Dissemination of Carrot Leaf Extract Feed Supplement Technology to Increase Laying Hen Productivity in Kecamatan Gumukmas. National Conference For Community Service. 7(2) : 403-407. [3] Central Statistics Agency. 2021. Carrot Production Growth in Indonesia. Indonesian Central Statistics Agency [4] Hadyarrahman Z, K.M. Yuliawati, and L. Syafnir. 2017. Influence of differences of leaves leaf extraction (Daucus carota L.) on antibacterial activity on Staphylococcus aureus and Escheriacia coli”, Prosiding Farmasi, 3(2): 634641 [5] Puspani, E., D.P.M.A. Candrawati and I.G.N.G. Bidura. 2018. The effect of aqueous extract of two leaves (Sauropus androgynus and Moringa oleifera) and its combination on egg production, yolk colour and yolk cholesterol levels in egg laying hens. International Journal of Multidisciplinary Approach and Studies Volume 05, (5) : 23-32. [6] Muchtadi, Tien R. "Sugiyono, and Ayustaningwarno, F. (2010)." Food science. World Journal of Biology Pharmacy and Health Sciences, 2025, 24(02), 533-538 538 [7] Siti, N.W., Bidura, I.G.N.G., Mayuni, S.N., Suasta, I.M., Wibawa, A.A.P.P. & Utami, I.A.P., 2019. Effect of Moringa oleifera leaf powder in diets on feed digestibility and external egg quality characteristics in laying hens. International J. Fauna Biological Studies 6(4), 113-118. [8] Steel, R. and J. Torrie, "Principles and procedures of statistics"New York, Toronto, London: McGraw-Hill Book Company, Inc. xvi + 481 pp. 1960 [9] Hy-Line. 2019. Management guide. West Des Moines [10] Hanapis, E.J. Guntoro, and Aswana. 2020 The effect of partially replacing commercial feed with market waste carrot flour up to a level of 12% on the weight of quail eggs (Coturnix-coturnix japonica). Stock Peternakan Journal. 2(1):16-25 [11] Hadrawi, J., S.P. Pitres, and Basri. 2022. The effect of moringa leaf powder supplementation on the production performance and egg quality of laying hens. Journal of Animal Science and Technology. 3(2): 43-48 [12] Ginting, N.R. 2025. The Effect of Carrot Leaf Extract (Daucus carota L.) on the Chemical Quality of Laying Hen Eggs. Thesis. Faculty of Animal Husbandry, Jember State Polytechnic. [13] H. Lukman. and Suryono, S., 2020. Characteristics of egg white and yolk pH, fat content, and organoleptic value of duck eggs injected with garlic (Allium sativum, Linn.) solution. Scientific Journal of Animal Science, 23(1), 16– 21. [14] Rahmawati, N., and A.C. Irawan. 2021. The effect of adding herbafit to feed on the physical quality of laying hen eggs. Journal of Tropical Animal Nutrition. 4(1):1-14 [15] Siti, N.W. dan I.G.N.G. Bidura. 2022. Effects of carrot leaves on digestibility of feed, and cholesterol and betacarotene content of egg yolks. South African Journal of Animal Science, 51 (6): 786-792 [16] Teteh, A., E. Lawson, K. Tona, E. Decuypere, and M. Gbeassor. 2013. Moringa oleifera leave: Hydro-alcoholic extract and effects on growth performance of broilers. International J. Poult. Sci. 12(7), 401-405. [17] Hammershoj, M., U. Kidmose, and S. Steenfeldt. 2010. Deposition of carotenoids in egg yolk by short-term supplement of coloured carrot (Daucus carota) varieties as forage material for egg-laying hens. J. Sci. Food. Agric. 90(7), 1163-1171.