Evaluation of Moringa oleifera leaf meal utilization as soybean meal protein substitution in rations to produce functional quail eggs

Rafidatun Nisa¹, Sumiati² and Tuti Suryati³

¹ Graduate School of Animal Nutrition and Feed Science, IPB University Department of Animal Nutrition and Feed Technology, Faculty of Animal Science, IPB University, Bogor 16680, Indonesia
sumiati@apps.ipb.ac.id
² Department of Animal Nutrition and Feed Technology, Faculty of Animal Science, IPB University, Bogor 16680, Indonesia
³ Department of Animal Products Technology, Faculty of Animal Science, IPB University, Bogor 16680, Indonesia

Abstract

The purpose of this study was to evaluate the effect of utilizing Moringa oleifera leaf meal as a substitute for soybean meal protein in the diet to produce functional quail eggs. This study used 240 quails aged 6-14 weeks. A completely randomized design (CRD) with 4 treatments and 5 replicates was used in this study. Treatments consisted of P0: Control ration, P1: Moringa oleifera leaf meal protein replaces 10% soybean meal protein, P2: Moringa oleifera leaf meal protein replaces 20% soybean meal protein, P3: Moringa oleifera leaf meal protein replaces 30% soybean meal protein. Data were analyzed using analysis of variance (ANOVA), if there were significantly different, Tukey test was conducted. The results showed that the use of moringa oleifera flour had no significant effect on egg performance and physical quality, but had a significant effect (p<0.05) on yolk color score, vitamin A, cholesterol content, yolk MDA and antioxidant activity. In conclusion, the utilization of moringa leaf meal as a substitute for soybean meal protein up to 30% in the diet increased yolk color score, vitamin A content and antioxidant activity, decreased cholesterol, as well as egg yolk MDA content, without affected performance and physical quality of eggs.

Keywords: egg quality, functional egg, moringa oleifera, quail, vitamin A

Introduction

Quail eggs are one of the livestock commodities that have a comprehensive nutritional content, making them very popular among the public as an economical and easily accessible source of animal protein. Apart from being a source of protein, eggs also contain various important nutrients such as vitamins, minerals, amino acids, essential fatty acids, phospholipids, lutein, as well as various bioactive components, antioxidants, and choline (Sanlier and Ustun 2021). Components in quail eggs that are considered very necessary in people's body resistance, namely: vitamin A and antioxidants. Therefore, it is necessary to utilize quail eggs as a food source of vitamin A and antioxidants.

Vitamin A is essential for human endurance and vision. Vitamin A deficiency can cause visual impairment, decreased immunity, and increased risk of infection (Waskitoningtyas and Pratama 2021). Likewise with the antioxidants present in quail eggs. Antioxidants in eggs also play an important role in maintaining health and reducing the risk of chronic diseases such as heart disease and cancer (Prihayati 2014). Therefore, it is important to maintain the vitamin A and antioxidant content in quail eggs by providing quality feed such as soybean meal.Soybean meal is the main protein source feed for poultry because of its complete amino acid content (Ispitasari and Haryanti 2022). The protein in soybean meal can increase egg production (Fadillah 2022). However, the high price of soybean meal due to imports is an obstacle (Santi 2017). To overcome this problem, local feed is needed that can replace soybean meal. Moringa leaves are one of the potential local feed alternatives.

Moringa leaves are a medicinal plant rich in antioxidant compounds such as flavonoids and β- carotene (Satriyani 2021). Moringa leaf flour can serve as a β-carotene supplement, producing antioxidant-rich quail eggs (Prihayati 2014). The vitamin A content in moringa leaves is higher than carrots, and contains phytochemicals such as flavonoids, saponins, tannins, and phenolic compounds with antimicrobial activity (Bukar et al 2010). Flavonoids can slow down osteomalacia, lower blood cholesterol, and increase high density lipoprotein HDL. Saponins have anticancer, antimicrobial, and cholesterol-lowering properties, while tannins can be used as antiviral, antibacterial and antitumor (Yildirım and Kutlu 2015). Research by Satria et al (2016) reported that the addition of 2% moringa flour in feed can increase consumption and egg weight of chickens and reduce feed conversion and yolk cholesterol. Bidura et al (2020) also reported that the use of 4%-6% moringa flour in laying hen feed increased calcium and β-carotene content in egg yolks and reduced egg yolk cholesterol. Therefore, this study aimed to evaluate the utilization of moringa leaf meal as a substitute for soybean meal protein in feed to produce functional quail eggs.

Materials and methods

Making Moringa oleiferaleaf flour

The process of making modified Moringa oleifera leaf flour from Siti et al (2019) begins with the separation of dark green leaves from the twigs, aerated at room temperature for 2 days, then dried in an oven at 55^oC for 24 hours. After that, the leaves were ground using a blender to form Moringa oleifera leaf flour (mash). Nutrient and phytochemical content of Moringa oleifera leaf flour (Table 1).

Research design and feed

The feed was prepared based on Leeson and Summers (2005) and adjusted to the nutritional requirements of laying quail. The research feed ingredients (Table 2) were then mixed until homogeneous. A total of 240 quails aged 6 were divided into four treatment groups, with each group consisting of five replications and twelve quails per replication and were placed in a colony cage. Maintenance was preceded by a two week adaptation period using multilevel colony cages. Cages were placed according to randomization. Treatments consisted of P0: Control feed, P1: Moringa oleifera leaf meal protein replaces 10% soybean meal protein, P2: Moringa oleifera leaf meal protein replaces 20% soybean meal protein and P3: Moringa oleifera leaf meal protein replaces 30% soybean meal protein. The observed variables were quail performance, eggs physical quality, yolk antioxidant activity, yolk malondialdehyde (MDA), cholesterol and vitamin A content of the yolk eggs produced.

Data analysis

The experimental design used a completely randomized design (CRD) with 4 treatments and 5 replicates. Data were analyzed by analysis of variance (ANOVA). If the treatment had a significant effect (p<0.05), it was continued with the Tukey test using SPSS 25 software. Vitamin

A content of eggs was analyzed descriptively.

Results and discussion

Quail performance

The performance of the research quails fed the treatments from 8-14 weeks of age is presented in Table 3.

Based on the results of ANOVA analysis of variance in Table 3, it shows that the utilization of moringa leaf meal as a substitute for soybean meal protein in feed has no significant effect on feed consumption, egg production, egg weight, egg mass production and feed conversion. Quail feed consumption in this study was 20.49-21.15 g head^-1 day^-1. This is due to the nutrient content in the treatment of moringa flour addition with the control ration is relatively the same so that nutrient consumption is also the same, especially energy consumption. Irawan et al (2012) stated that basically quail will consume feed to fulfill their energy. Wahju (2004) also states that quail will consume feed until their energy is fulfilled. The addition of moringa flour in feed can increase feed consumption compared to the control treatment. Feed consumption in this study was higher than the research of Khoir et al (2021) with feed consumption of 20.03-20.87 g head^-1 day^-1. Feed consumption that is not significantly different in each treatment makes protein consumption in each treatment relatively the same. Protein is an important nutrient for the body because it consists of amino acids that cannot be synthesized by the body. Amino acids available in the feed will be used to synthesize skeletal muscle or build body tissues, meet production needs and the excess will be converted into energy (Aziz et al 2020). Lisnahan et al (2018) also explained that amino acids play an important role in the body's metabolism to maintain health, growth and development of reproductive organs. According to Kashavarz (2003) variations in protein consumption will have an impact on egg production, egg weight and egg mass produced.

Another factor that causes feed consumption is the temperature of the cage environment. The average temperature of the cage environment at the time of the study ranged from 26-28^oC.

Laying poultry can maintain stable production in temperatures of 10-30ºC (Suprijatna et al 2009), while the ideal air humidity for quail is between 30-80% (Wuryadi 2013). According to Ustomo (2016) the effect of Heat Stress Index (HSI) on performance is: HSI <150 does not cause performance problems; HSI 155 is the upper limit of performance decline; HSI 160 causes a decrease in feed intake, an increase in water intake, and a decrease in performance; HSI 165 triggers early death and permanent damage to the lungs and circulatory system; and HSI 170 causes high mortality. During the rearing period the average HSI was 128.46.

Physical quality of eggs

The physical quality of the research quail eggs fed the treatments from 8-14 weeks of age is presented in Table 4.

Based on the results of ANOVA analysis of variance in Table 4, it shows that the utilization of moringa leaf meal as a substitute for soybean meal protein in feed has no significant effect on egg white, egg yolk, shell, shell thickness, haugh unit (HU), egg index, egg white index and egg yolk index but significantly different (p<0.05) on egg yolk color score. This is because the type of feed and the level of feed consumption given moringa leaf flour with the control treatment are similar, resulting in egg weights that are not much different. The increase in overall egg weight is influenced by white and yolk weights. According to Burlando et al (2010) Moringa leaves contain methionine which functions as an essential amino acid that greatly affects egg weight and will certainly affect the increase in protein content in eggs. Methionine amino acids are superior to other amino acids in increasing egg weight (Leeson and Summers 2001). Methionine acts as a methyl donor, helping metabolize protein, chlorine and carbohydrates in the body. Calcium and phosphorus in feed play an important role in the formation of a good egg shell. The calcium content in moringa flour given did not increase the thickness of quail egg shells. According to Achmanu et al (2011) the thickness of the egg shell is closely related to the level of calcium it contains.

Utilization of moringa flour in feed can increase the yolk color score compared to the control treatment. This is influenced by the content of carotenoid compounds contained in moringa flour.

The more the use of moringa flour in feed, the intensity of egg yolk color increases. According to Sujana et al (2006) that feed will affect the color of egg yolk if the feed ingredients used contain carotenoid pigments such as β-caroteneand xanthophyll. β-carotenecontained in Moringa leaves reaches 47.110 μg/g (Winarno 2003). This study showed that the utilization of moringa flour as a substitute for soybean meal protein at the 30% level was the best level to produce egg yolk color scores. The yolk color score in this study was lower than the research of Paryanta et al (2019), which was 7.86-8.09 by giving drinking water with moringa leaf solution, it was thought that this was due to the use of moringa leaves at a lower level than the research of Paryanta et al (2019). According to Sudaryani (2003) the color of egg yolks affects consumer tastes where in general consumers prefer egg yolks with a concentrated color.

The HU value,egg index, egg white index and yolk index that were not significantly different were probably caused by the nutrient content in the feed. In this study, the protein content containing moringa flour decreased from the control treatment, so the egg index, egg white index and yolk index did not increase. In addition, the content of tannin compounds in the treatment ration also affects egg weight. According to Hagerman (2002) tannins have properties as protein precipitators so that the availability of protein decreases and results in egg index, egg white index and yolk index decreasing. According to Tugiyanti and Iriyanti (2012) egg quality can be measured based on the HU value which is determined based on albumen height.

HU value is the logarithm of egg white height which is then correlated with egg weight, if the difference in egg weight is not significantly different then the difference in HU is also not significantly different. In this study, feed protein containing moringa flour tended to be lower.

However, this tendency did not cause the HU value to decrease. According to USDA (2000), HU values are classified into four grades: AA quality for eggs with HU values >72, A quality for HU values between 60-72, B quality for HU values between 31-60, and C quality for HU values <31.

In this study, the HU value obtained was included in the AA quality category with a HU value of >72 which indicates good egg quality. According to Kashavarz (2003) variations in protein consumption both increase and decrease will have an impact on the weight of the eggs produced. The higher the egg weight the greater the egg index obtained. According to Argo et al (2013) the egg index is influenced by the content of protein, fat, and essential amino acids in the feed, and other factors that affect the egg index include egg size, length of storage, temperature, vitelin membrane quality, and feed nutrition.

Chemical quality content of egg yolk

The chemical quality of egg yolks of research quails fed the treatment diet from 8-14 weeks of age is presented in Table 5. Based on the results of ANOVA analysis of variance in Table 5, the treatment of moringa leaf meal as a substitute for soybean meal protein up to 30% in feed resulted in a significant decrease (p<0.05) in cholesterol content, egg yolk MDA, antioxidant activity measured by the IC50 method and increased the vitamin A content of quail eggs.

The results showed that the highest vitamin A content was found in the P3 treatment with the utilization of moringa flour as a substitute for soybean meal protein up to 30% in feed, which amounted to 2376 IU/100 g. The data showed an increase in vitamin A egg yolk by 76% (Figure 1) when compared to the control ration. This is due to the content of moringa leaf meal which can increase vitamin A in egg yolk. The use of moringa flour which is high in vitamin A in feed acts as an antioxidant. Vitamin A content in egg yolk is influenced by β-carotene intake. β-carotene is a type of carotenoid that acts as pro-vitamin A, which will be converted into vitamin A in the intestinal mucosa and absorbed in the form of vitamin A. Leeson and Summers (2001) stated that the vitamin A content of egg yolk can increase with increasing provitamin A content in the diet. The vitamin A content of egg yolk in the control treatment had a smaller vitamin A content than the other treatments.

The lowest cholesterol content was found in the P3 treatment with the utilization of moringa flour as a substitute for soybean meal protein up to 30% in the feed, namely 433.38 mg/100 g, here there was a 16% decrease in cholesterol (Figure 1) when compared to the control ration.

Carotenoids are one of the active substances in moringa flour that can help reduce cholesterol content in quail eggs. This is in line with the opinion of Olugbemi et al (2010) that moringa leaves have a hypocholesterolemic effect that not only lowers cholesterol in eggs but also improves the nutritional quality of eggs including the content of vitamin A. The results of this study indicate that the higher the content of moringa leaf flour in the ration, the smaller the cholesterol content of egg yolk. The low cholesterol content in the P3 treatment may also be due to the high crude fiber content in moringa flour.

Based on the results of the malondialdehyde(MDA) content test, egg yolks treated with moringa leaf flour as a substitute for soybean meal protein up to 30% showed lower egg yolk MDA results compared to the control treatment. Egg yolk MDA levels of the control treatment had a value of 0.60 mg/kg, while the P3 treatment amounted to 0.40 mg/kg. The lower yolk MDA levels may be due to the high antioxidant activity of moringa flour in the feed. MDA is an indicator that is often used to measure lipid peroxidation in the body which is closely related to oxidative stress (Ayala et al 2014). The high concentration of MDA indicates the oxidation process that occurs in the cell membrane. Utilization of moringa flour as a substitute for soybean meal protein up to 30% can reduce egg yolk MDA levels by 33%. This indicates that the provision of moringa leaf meal has the potential as an antioxidant to reduce the effects of oxidative stress and improve health in quail.

Flavonoids are active substances contained in Moringa leaves that have the potential to increase antioxidant activity in quail. The role of antioxidants is to convert free radicals into more stable compounds. Moringa leaf flour can increase antioxidant activity in eggs as indicated by a lower IC50 value. The lower the IC50 value of an ingredient, the higher the antioxidant activity (Molyneux 2004). Utilization of moringa leaf meal as a soybean meal protein substitution up to 30% can reduce the antioxidant activity of egg yolk IC50 by 77% (Figure 1).

Figure 1. Percentage increase in vitamin A as well as decrease in cholesterol content, egg yolk MDA and antioxidant activity IC50 of quail eggs in the study

Conclusion

The use of Moringa oleifera leaf meal as a substitute for soybean meal protein in feed up to 30% improved yolk colour score, vitamin A content and antioxidant activity of eggs, and reduced cholesterol and MDA levels of egg yolk, without compromising the performance and physical quality of quail eggs.

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