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The Effect of Carot Leaves Meal Fermented in Diets on Egg Production, Yolk Cholesterol and Beta-Carotene in Yolk of Hens
I Gusti Nyoman Gde Bidura, Ni Wayan Siti, Anak Agung Putu Putra Wibawa, I Nyoman Tirta Ariana and Eny Puspani
Faculty of Animal Science, Udayana University, Denpasar-Bali, Indonesia E-mail: [email protected]
ABSTRACT
The aim of this study was to examine the effect of carrot (Daucus carota) leaves fermented with Saccharomyces spp.
(fermented Carrot leave meal/FCL), on the content of cholesterol and β-carotene in the yolk and the performance of egg production in laying hens. Before being added to the diet, carrot leaf flour was fermented with Saccharomyces spp. culture.
(3.106 cfu/g) with 3 days of fermentation. Two hundred and eighty-eight laying hens aged 40 weeks were randomly assigned to 4 treatment diets. Dietary treatments were as follows: 0%, 2%, 4%, and 6% FCL levels as a feed ingredient in diets. The diets formulation based on equal amounts of protein and energy. The results showed that 2-6% FCL in the diet, significantly (P<0.05) increased egg production, egg weight, and feed efficiency compared to control. Yolk color, weight, and concentration of β-carotene in the yolk increased significantly (P<0.05) in the FCL treatment group, while the yolk cholesterol concentration decreased (P<0.05) than control . It can be conclusion that provision of 2-6% FCl in the feed can increase hen-day production and the β-carotene in yolk, while the cholesterol content in the yolk decreases.
Key words:
Carrot leaf waste, yolk color, shell thickness, hens
Introduction
Since the use of antibiotics in poultry feed is prohibited to increase the quantity and quality of poultry products, researchers have begun to focus on the use of phytogenic compounds as non-antibiotic growth promoters. There are many studies investigating herbal plants as growth promoters, improving livestock health, and quality of poultry production [1,2,3,4,5,6,7]. According to [8], the general advantages of herbal medicines are effectiveness, product safety, acceptance by farmers, and affordability.
Generally, as a source of yolk color in feed for hens is yellow corn, because yellow corn contains high xanthophyll, lutein, and zeaxanthin dyes [9]. But, using of yellow corn as a feed ingredient still competes with human food needs.
Lack of carotenoid dyes can cause egg yolks to turn pale and this is not liked by consumers, because it looks unhealthy [10]. Therefore, it is necessary to find an alternative to corn feed with feed ingredients that are not competitive with humans.
Interesting to study is the post-harvest waste from carrot plants, namely carrot leaves. Carrots (Daucus carota L.), belonging to the Apiaceae family, are a major vegetable source of β-carotene (BC). β-carotene compounds in carrot leaves are effective in egg yolk pigmentation. The main carotenoid compounds in carrot leaves are carotenoids α- carotene, β-carotene, and β-cryptoxanthin [11]. In addition, the antioxidant properties of BC will be able to prevent egg damage, because storage of BC is in egg yolk [12], and BC can improve the immune system by increasing antibody responses in poultry.
Carrot plants cultivated throughout the world are root vegetables that are biochemically rich in sources of minerals, fiber, carbohydrates, flavonoid antioxidants, essential micronutrients, and BC [13]. Carrot production is quite high, that is, in every 1 ha of carrot plant, the actual productivity is 15 tons of carrot tubers and 5% of it as carrot leaf waste which is not used as human food [14]. The nutrient content of carrot leaf flour is quite good, namely 9.27% crude protein;
1.20% crude fat; and 2487 kcal/kg metabolized energy [15]. However, the weakness of carrot leaves is their high crude fiber content, which is 19.64%, so they need to be fermented before being given to poultry. The use of carrot leaf meal up to a level of 6% in the ration did not have an impact on feed consumption and feed efficiency. Yeast of Saccharomyces spp. was used as a fermentation inoculum, because it has been reported to be able to degrade crude fiber [16,17].
Since the use of antibiotics is prohibited to increase the quantity and quality of egg products, researchers are interested in phytogenics as non-antibiotic egg production promoters. The purpose of this study was to increase egg production, the content of beta-carotene in the yolk, and reduce the cholesterol content in the yolk in laying hens.
Material and Methods
Animals, diets and management. A total of two hundred and eighty-eight laying hens aged 40 weeks old laying hens of nearly similar initial body weight (1740.82±40.58 g) were assigned into 4 experimental groups each including 72 hens and maintained at individually cages of 35 x 40 x 40 cm. Carrot leaves (Daucus carota L.) were supplied from a carrot farm in the Baturiti area, Tabanan-Bali, Indonesia, after harvesting the fruit. The carrot leaves were cleaned of existing dirt, then cut into small pieces with a grass sieve machine, then dried in the sun. After drying, then finely ground with a diameter of 1-2 mm. Before being added to the diet, carrot leaf flour was fermented (FCL) with
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Saccharomyces spp. culture. (3.106 cfu/g) with 3 days of fermentation. Dietary treatments were as follows: 0%, 2%, 4%, and 6% FCL levels as a feed ingredient in diets. The diets formulation based on equal amounts of protein (CP 18%) and energy (ME: 2900 kcal/kg). All laying hens were fed 120 g per head per day in two doses (morning and evening) and drinking water was given ad libitum. Eggs were collected twice a day (morning and evening).
Table 1 Composition of feed ingredients and nutritional content calculation results in feed treatments for laying hens aged 40-50 weeks
Ingredients (%) Level of FCL in diets (%)
0 2 4 6
Yellow corn 53 51.8 50.8 49.6
Pollard 13 12 11.08 10.2
Soybean 14 14.1 14.1 14.35
Fish meal (CP 45%) 13.75 13.78 13.9 13.8
Coconut oil 0.53 0.53 0.45 0.4
FCL 0 2 4 6
NaCl 0.25 0.25 0.25 0.25
Mineral 5.47 5.54 5.42 5.4
Total 100 100 100 100
Composition*):
Metabolizable energy (ME) (Kcal/kg) 2900 2901 2901 2901
Crude protein (%) 18.02 18 18.01 18.01
Ether Extract (%) 6.87 6.83 6.71 6.64
Crude fibre (%) 3.2 3.47 3.75 4.05
Ca (%) 3.8 3.82 3.81 3.81
P (%) 1.43 1.44 1.44 1.44
Arginine (%) 1.31 1.3 1.29 1.28
Cysteine (%) 0.33 0.33 0.33 0.33
Lysine (%) 1.4 1.4 1.4 1.39
Methionine (%) 0.44 0.43 0.43 0.43
Tryptophan (%) 0.25 0.24 0.24 0.24
*) Calculations based on the nutrient composition table according to [18]
Egg performance and quality. Feed consumption was calculated every week, by means of the difference between the amount of feed provided and the remaining feed from each experimental unit (a total of 24 experimental units). Total feed consumption were expressed in g/head/70 days. Eggs were collected daily and egg counts were recorded to determine the average egg number (including abnormal eggs), which was represented as a percentage of the average number of hens laying eggs during the period (eggs/head/day). Eggs were weighed individually on an analytical balance with a sensitivity of 0.01 g. After weighing, the eggs were submitted for analysis of specific gravity (g/cm3).
Egg Multitester-EMT 7300 tool to find out HU, color, egg weight, and albumen height. Flat plane glass used as a base for laying egg fragments. This glass was used in measuring egg yolk height, albumen width, and yolk width using a tripod micrometer. Measurement of yolk color value using the "Roche yolk color fan" in the form of a standard color fan sheet with a score of 1-15 from pale to dark orange or dense. Cholesterol levels in serum and yolk were analyzed according to the procedure of [19] and the content of β-carotene in the yolks following the procedure of [20].
Result
There was no significant difference (P>0.05) in final body weight, total feed consumption, and egg weight between the treatment groups (Table 2). There was a significant increase (P<0.05) in egg number, hen-day production, total egg weight, egg mass, and feed conversion ratio (FCR) with FCL supplementation in the diet.
Table 2. The effect of FCL in ration on hens egg production in laying hens from 40-50 weeks of age
Variables Treatment Groups1
SEM2
A B C D
Initial body weight (g bird-1) 1738.16a3 1750.71a 1747.45a 1751.48a 39.692 Final body weight (g bird-1) 1808.31a 1836.18a 1850.72a 1871.93a 60.385 Feed consumption (g bird-1 /70 days) 7883.40a 7969.50a 8035.90a 7963.20a 98.463
Egg numbers (egg bird-1/70 days) 56.50b3 62.50a 62.58a 62.55a 0.0275
Hen-day production (%) 80.71b 89.29a 89.40a 89.36a 2.0727
Egg weight (g egg-1) 61.13a 61.43a 61.52a 61.49a 0.293
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Total egg weight (g bird-1/70 days) 3453.85b 3839.38a 3849.92a 3846.20a 98.459
Egg mass (g bird-1/weeks) 345.39b 383.94a 384.99a 384.62a 9.385
FCR (g of feed/g of egg) 2.28a 2.08b 2.09b 2.07b 0.042
Note:
1. Group of hens fed without FCL as control (A); 2% FCL (B); 4% FCL (C); and 6% FCL (D), repectively.
2. Standart error of the treatmen means.
3. Values with different letters in a row are significantly different (P<0.05)
Table 3 shows the egg quality parameters for chickens fed on feed containing fermented carrot leaf flour (FCL). There was no effect (P>0.05) in albumen weight and albumen percentage, specific gravity, Haugh unit, and egg shape between groups (A, B, C, and D). The eggshell thickness was increased (P<0.05) with FCL supplemented in the feed.
There was a significant difference (P<0.05) for yolk weight and yolk percentage, eggshel weight and eggshell percentage which were higher for diets containing FCL.
Table 3: Internal egg quality of hens fed supplemented with varied levels of FCL
Variables Treatment Groups1)
SEM2
A B C D
Eggshel weight (g) 6.31b 6.66a 6.81a 6.75a 0.107
Eggshell (%) 10.27b 10.74a 10.92a 10.81a 0.128
Yolk weight (g) 15.87b 16.60a 16.64a 16.86a 0.196
Yolk (%) 25.82b 26.75a 26.69a 26.98a 0.207
Albumen weight (g) 63.91a 62.51a 62.39a 62.21a 0.592
Albumen (%) 39.27a 38.78a 38.90a 38.87a 0.394
Specific gravity (g/cm3) 1.063a 1.065a 1.067a 1.066a 0.079
Shell thickness (mm) 0.389b 0.425a 0.427 0.429 0.0115
Haugh unit (albumen height:egg weight) 75.92a 76.39a 76.15a 75.99a 0.393 Egg shape (egg width/egg length) x 100% 75.37a 74.81a 74.95a 74.82a 0.794 Note:
1. Group of hens fed without FCL as control (A); 2% FCL (B); 4% FCL (C); and 6% FCL (D), repectively.
2. Standart error of the treatmen means.
3. Values with different letters in a row are significantly different (P<0.05)
Table 4 shows the concentration of yolk cholesterol and β-carotene in yolk, yolk color, and blood lipid profile from laying hens feed containing fermented carrot leaf flour (FCL).. There was no significantly different (P>0.05) in trigliceride, hight lipoprotein density (HDL), and low lipoprotein density (LDL) in serum of hens between treatments (A, B, C, and D). The use of FCL (B, C, and D) in the diet significantly (P<0.05) increased the yolk color score and the concentration of beta-carotene in the yolk, but decreased the cholesterol content in the serum and yolk. In more detail, yolk color and BC content in the yolk were presented in Figures 1 and 2.
Table 4. Concentration of cholesterol and β-carotene in yolk from laying hens feed FCL meal
Variables Treatment Groups1
SEM2
A B C D
Yolk color (1-15) 6.85b 8.92a 9.17a 9.06a 0.417
β-carotene (µg kg-1) 1.039b 1.285a 1.304a 1.427a 0.0591
Yolk cholesterol (mg/100 g) 11.85a 10.39b 10.25b 10.17b 0.352
Blood lipid profile (mg dl-1)
Total cholesterol 148.05a 132.71b 128.03b 125.95b 4.097
Triglyceride 27.04a 25.84a 26.16a 25.29a 2.583
HDL 98.17a 96.52a 96.93a 97.65a 6.048
LDL 39.07a 38.21a 35.74a 36.41a 5.29
Note:
1. Group of hens fed without FCL as control (A); 2% FCL (B); 4% FCL (C); and 6% FCL (D), repectively.
2. Standart error of the treatmen means.
3. Values with the same letter in one row, showed not significantly different (P>0.05).
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0 1 2 3 4 5 6 7 8 9 10
A (control) B (2% FCL) C (4% FCL) D (6% FCL)
Yolk color (1-15)
0 0.2 0.4 0.6 0.8 1 1.2 1.4 1.6
A (control) B (2% FCL) C (4% FCL) D (6% FCL)
β-carotene (µg kg-1)
Figure 1. Effect of FCL supplementation in diet (groups B, C, and D) on value of yolk color (1-15) and yolk beta- carotene content in laying hens.
Discussion
Performance of laying hens (egg numbers, egg mass, and feed efficiency) increased with FCL supplementation in the diet, while feed consumption did not show any difference between treatments. Herbal leaf fermentation can improve the quality of herbal leaves and has no effect on feed intake in chickens [3,21,22]. The production and quality of chicken eggs increased with the supplementation of herbal extracts without or fermented through drinking water or mixed in the diet [1,2,3,21,23]. The increase in egg performance was due to the content of phytochemical compounds in carrot leaves, such as: saponins, flavonoids, and tannins [24]. In addition, β-carotene compounds in FCL can be converted into vitamin A which plays a role in epithelial cell differentiation and maintains digestive organs, so that it will affect the digestibility and absorption of nutrients [25]. The inclusion of carrot food in the feed can increase the value of retinol and antioxidant compounds [26].
FCL supplementation in the diet can increase eggshell percentage, yolk percentage, and shell thickness. The results of the research of [20] showed that increasing the application of Moringa leaf flour in the diet significantly increased egg production, egg mass, Mg and Ca content in egg shells. The research of [25] found that supplementation of carrot waste flour in broiler rations up to 6% had the same effect on the calcium mass of meat, but decreased the mass of protein in the meat. In line with the research of [27], that supplementation of 1-8% carrot leaf powder in the feed did not significantly affect all external egg quality variables (albumin index, egg weight index, HU, egg yolk index, and chicken egg yolk color). Similar to [28] reported that herbal flour in hen diets did not produce a difference in the percentage of albumen. Similar to [29] that the use of 0.8% carrots in a diet containing sorghum had no effect on the specific gravity of eggs. The level of protein in the feed greatly affects the external quality of eggs [30]. Increased nutrient intake due to fermented carrot leaves. The nutritional value of the feed increases with the fermentation process [16]. Increased digestibility of feed in chickens can affect protein synthesis and calcium mineral intake which can affect protein mass and calcium mass in chicken products [25]. The range of Haugh units (white height: egg weight) of eggs in this study was between 75-76.4. The Haugh unit in this study is still lower than that of [27] who reported that supplementation of 1-4 g of Carrot leaf powder in feed resulted in Haugh units greater than 79. Hammershøj et al. [11]
reported that carrot supplementation in the diet reduced egg weight, but the percentage of yolk was not different compared to the control (without carrot supplementation).
The use of colored carrots as forage in laying hens feed can increase the total carotenoid concentration in egg yolks to 25-75% higher than chickens not fed carrots [11]. Beta-carotene has an accumulation rate of less than 1% in egg yolks.
Some researchers reported that there was a linear increase in the amount of retinol in the yolk, due to an increase in the amount of BC in the feed [20,31]. BC content in egg yolk ranged from 1.07-2.12 μg/kg [32] and 0.16-1.62 μg/kg [33].
The increasing number of BC in the feed as a result of the use of FCL in the diet will have an effect on increasing the yellow color and beta-carotene content in the yolk [20]. According to [34], carrot leaves contain beta-carotene as much as .2-6.1 mg/100g fresh weight, while yellow corn contains: 0.051 mg/100g. The yolk color increased by addition of 0.8% carrot flour in the feed [26].
Provision of 70 g of carrot flour per day into the ration of laying hens has been shown to effectively increase the color value of egg yolks, especially the content of lutein, alpha-carotene and BC [11]. Inhibition against Escherichia coli and Salmonella sp. really depends on the type of plant herbs and the concentration of herbal extracts used [35].
Cholesterol content in yolks can be suppressed by beta-carotene compounds contained in herbal leaves (Moringa,
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Sauropus) [20]. According to [36], the presence of BC contained in Noni herbs can reduce cholesterol levels in the blood, because BC can inhibit the action of the HMG-CoA reductase enzyme which plays a role in the formation of mevalonate in cholesterol biosynthesis. Suppression of mevalonate inhibits the formation of squalene and lanosterol, both of which are involved in cholesterol biosynthesis through a series of reactions. Teteh et al. [37] reported that the decrease in fat and cholesterol content in egg yolks was caused by increased levels of saponins contained in herbal leaves, as antinutrients that can reduce digestion and lipids. Fermentation of herbal leaves (Sauropus) can increase the nutrition of herbal leaves and at the level of 14% fermented herbal leaves in the ration produces the lowest cholesterol levels [38]. According to [39], the use of carrot flour in the diet can activate the calcium activated neutral protease (CANP) enzyme and at the level of 2-6% carrot waste flour in broiler feed, has not had a significant effect on carcass fat and carcass skin pigmentation. Pambayun [40] reported that phenolic compounds in herbal extracts played a role in inhibiting the growth Terpenoid compounds can be antibacterial by damaging bacterial cell membranes [41].
Conclussion
It was concluded that supplementation of 2-6% fermented Carrot leaf meal in diet laying hens increased egg numbers, hen-day production, total egg weight,egg mass, feed efficiency, yolk percentage, eggshell percentage, yolk color, and concentration of β-carotene in the yolk. On the other hand, it significantly reduced cholesterol in the yolk.
Acknowledgements
The authors thank technician Mr. Kapong for hen management, technician Sagung Ngurah Mayuni and Emy Suastini for cholesterol and beta-carotene analysis, and Mr. Lecir for carrot supply. Thanks also to the Chancellor of Udayana University, Denpasar Bali, Indonesia for the funds provided, so that this research can be completed.
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