CC BY
3JWPR
Journal of World's Poultry Research
2021, Scienceline Publication
J. World Poult. Res. 11(3): 293-301, September 25, 2021
Research Paper, PII: S2322455X2100035-11 License: CC BY 4.0
DOI: https://dx.doi.org/10.36380/jwpr.2021.35
Effects of Different Levels of Moringa oleifera Whole Hydroalcoholic Extract and Seed Powder on the Hatching Rate, Nutritional Value, and Immune Response of Chukar Partridge Eggs
Hassan Habibi \ Mohammad Amin Kohanmoo2, and Najmeh Ghahtan3
'Department of Animal Sciences, Faculty of Agricultural and Natural Resources, Persian Gulf University, Bushehr, Iran 2Department of Plant Breeding, Faculty of Agricultural and Natural Resources, Persian Gulf University, Bushehr, Iran 3Department of Medicinal Chemistry, Faculty of Chemistry, Shiraz University of Technology, Shiraz, Iran
Corresponding author's E-mail address: [email protected]; ORCID: 0000-0001-8162-6205
Received: 08 June 2021 Accepted: 05 August 2021
ABSTRACT
The present study aimed to investigate the effect of different levels of Moringa oleifera whole seed powder (MOWSP) and whole seed hydroalcoholic extract (MOWSE) on biochemical factors including minerals, fatty acids profiles, Haugh units, cholesterol content, immune response, and hatchability rate of the eggs of Chukar partridge. A total of 225 Chukar partridge were randomly divided into five groups with three replicates of 15 birds in each group. The MOWSP was provided as a supplement at the rates of 0 g (control), 5 g, and 10 g per each kg of a diet and MOWSE at the rates of 0.5 % and 1% in drinking water. Hatchability rate and Haugh unit were, respectively, increased and decreased in all treatments in comparison with the control group. The highest and the lowest hatchability rates were recorded in the MOWSE-1% and MOWSE-0.5% supplemented groups, respectively. Birds fed with MOWSE-1% had significantly higher Iron levels than birds fed with the control diet. However, copper, zinc, and magnesium levels in the Chukar partridge eggs had no significant change, compared with the control group. Further, the C18:1, C17:0, and C16:0 of eggs were increased in response to the increase of dietary MOWSP supplementation, however, proportions of C18:0 and C18:2 decreased. It was also found that MOWSE-1% increased the antibody titers of Newcastle Disease vaccine on 69 days and MOWSP-1% and MOWSE-1% increased the titers of Avian Influenza on 59 days. It was concluded that 1% of MOWSP or MOWSE is a beneficial additive for Chukar partridge.
Keywords: Alectoris chukar, Cholesterol, Fatty acids profiles, Hatchability, Minerals
By 2050, the world's population will reach 9.1 billion, which is 34% more than today. Annual meat production will need to rise by over 200 million tons to reach 470 million tons (FAO, 2009). The Chukar partridge (Alectoris chukar, Aves: Galliformes, hereafter, chukar) has a very wide distribution, ranging from Eurasia, China, and Mongolia in the east, to southeastern Europe in the west (Habibi et al., 2019a). The Chukar is one of the most important game birds (Barbanera et al., 2007). Chukar partridges are exploited for hunting, meat, and egg production (Pourghanbari et al., 2016). Partridge eggs are an excellent source of nutrients for humans due to having unsaturated fatty acids, low cholesterol levels, and high levels of minerals (Réhault-Godbert et al., 2019).
However, there has been little research on the quality of chukar partridge eggs. Therefore, providing an effective and practical strategy to increase egg hatchability can be crucial in terms of production (Alikwe and Omotosho, 2013; Ahmad et al., 2017; Ahmad et al., 2018). Various factors can affect egg hatchability and biochemical properties such as age, genetics, gender ratio, storage period, feeding, weight of breeder animals, and egg weight (Caglayan, 2014).
Proper nutrition and the use of beneficial supplements in the diet of laying birds have been reported as main factors affecting the quality of eggs and hatchability rate (Habibi et al., 2019). In recent decades, an intensive amount of research has been focused on the development of natural growth promoters to enhance poultry production by stimulating their immune system,
TBBTSKBUjliBfflSri Habibi H, Kohanmoo MA, and Ghahtan N (2021). Effects of Different Levels of Moringa oleifera Whole Hydroalcoholic Extract and Seed Powder on the Hatching Rate, Nutritional Value, and Immune Response of Chukar Partridge Eggs. J. World Poult. Res., 11 (3): 293-301. DOI: https://dx. doi.org/10.36380/jwpr.2021.35
reducing feed costs, and increasing weight gain. Restriction on the use of man-made antibiotic growth promoters has created the need for comprehensive research on potential alternatives. One possible alternative is phytogenic feed additives (Windisch et al., 2008). In contrast to synthetic feed additives, phytogenic additives are more favorable to customers and clear up health concerns since they are safe and eco-friendly (Imoleayo Sarah Oladeji et al., 2019). A traditional source of nutritional supplements is Moringa medicinal tree. There are 13 species in Moringa genus among which the Moringa oleifera (M. oleifera) is the most studied and long-term used species (Nur Zahirah Abd Rani et al., 2018; Abdulkarim et al., 2005; Alikwe and Omotosho, 2013). M. oleifera also known as the drumstick tree grows in semiarid, tropical, and subtropical areas and is used for several purposes (Worku, 2016).
M. oleifera leaf (MOL) contains 25-27% crude protein (Gadzirayi et al., 2012) and high amounts of minerals and vitamins. The protein quality of MOL has been reported to be comparable to that of milk and eggs (Castillo, 2018). Chemical analysis of M. oleifera seed has revealed circa ether extract, dry matter as well as ash, crude protein, crude fiber, and has been applied in animal diets (Mabruk et al., 2014, Ayasan, 2015; Ahmad et al., 2017). M. oleifera seeds have been reported to be a good source of proteins, minerals, and fats (Compaore et al., 2011). Several studies have been performed to assess the effects of M. oleifera supplementation in broiler and layer chickens (Nkukwana et al., 2014; Mabusela et al., 2018). However, there is no literature for evaluation of the effects of M. oleifera supplementation on hatchability rate and egg nutritional components. Therefore, this study was conducted to investigate the effects of M. oleifera supplementation on hatchability rate, immune response, and egg quality.
MATERIALS AND METHODS
Experimental birds
A total of 225 (seven-month-old) chukar partridges were randomly divided into five groups with three replicates of 12 females and 3 males. The average body weight did not differ between the groups at the beginning of the trial period. All birds were allowed to adapt for a period of seven days, consuming an ad libitum commercial diet for laying partridge (Table 1). Strict sanitation practices were maintained in the facility throughout the experiment. The cages were cleaned daily to reduce the probability of any disease outbreak. Vaccinations and
medications were imposed when deemed important during the experimental period. The control group (group T1) was fed the same diet throughout the experiment. The remaining four groups were fed the control diet with supplementation with M. oleifera whole seed powder (5 and 10 g/kg for group T2 and T3, respectively) and 0.5% and 1% M. oleifera whole seed hydro-alcoholic extract in drinking water for groups T4 and T5, respectively.
Table 1. Composition of basal diet1
Ingredient g/kg
Corn 518.00
Soybean meal 355.00
Soybean oil 31.40
Dicalcium phosphate 7.00
Limestone 75.00
Sodium chloride 2.80
Sodium bicarbonate 1.00
L-Lys-HCl 1.30
DL-Met 3.40
Vitamin and mineral premix1 5.00
Phytase 10000 0.10
Total 1000.00
Analysis
Metabolizable energy, Kcal/kg 2800.00
Crude protein 19.84
Calcium 3.10
Available phosphorous 0.32
Sodium 0.15
Chloride 0.23
Lysine 1.08
Methionine 0.48
Methionine + Cysteine 0.88
Threonine 0.65
Tryptophan 0.22
Arginine 1.26
Isoleucine 0.77
Valine 0.83
Provided the following per kg of diet: Vitamin A, 10000 iu; Vitamin D3, 4500 IU; Vitamin E, 65 Iu; Vitamin K3, 3 mg; Vitamin B1, 2.5 mg; Vitamin B2, 6.5 mg; Vitamin B3, 60 mg; Vitamin B5, 18 mg; Vitamin B6, 3.2 mg; Vitamin biotin, 0.22 mg; Folic acid, 1.9 mg; Vitamin B12, 0.017 mg; Choline chloride, 1400 mg, Mn, 120 mg; Zn, 110 mg; Fe, 20 mg; Cu, 16 mg; I, 1.25 mg; Se, 0.3 mg
Preparation of Moringa seed powder
The seeds of M. oleifera were harvested from fully grown Moringa trees in Bushehr Province of southern Iran. Afterward, the seeds were dried, ground, and added to the diet.
Extraction of Moringa seed extract
Hydro-alcoholic extract (ethanol 70%) was prepared by seed soaking for 48 hours at room temperature and then filtered with filter paper.
Analysis of minerals
Minerals were evaluated using plasma atomic emission spectroscopy (ICP-AES, OPTIMA 5300DV, PerkinElmer, Waltham, MA) as a formerly reported procedure. In brief, 400 mg of the seed powder was weighed into a beaker and was digested in 4 mL of HNO3-HClO4 (4:1). Then it was heated to get dry. The residue then was treated with 0.1 N HNO3 and its volume was increased to 25 mL with double-distilled water. Certified standard minerals were applied for the determination of the elements (AOAC, 1990).
Cholesterol assay
Egg collection from adult chukar was performed daily (since seven months of age). In order to measure the cholesterol, 1 g of yolks was added to 9 mL water containing 2% NaCl and was kept in a shaking rotary for 2 hours. Subsequently, 1 mL of the diluted yolk was diluted 10 times. Then, 10 ^l of the sample was added to 100 ^l of salt solution and 1 mL of the enzymatic reagent. Standard cholesterol also passed the same steps. Samples were kept in a water bath at 37° C for 15 minutes, and the light absorbance of the samples was measured at 500 nm wavelength. 10 ^l of deionized water was used as a blank sample (Behnamifar et al., 2015).
Haugh unit
Haugh unit (HU) values were calculated using the following formula (Aboonajmi., 2010):
HU =100 Log (H+7.57-L7XW0'37)
Where, H is albumen height in millimeters and W denotes egg weight in gram.
Hatchability
All the experimental groups were placed into an incubator on the same date. The setter part of the incubator was set at 37.8°C and 55% RH, and eggs were automatically turned every hour. On day 20 of incubation, all the experiment eggs were transferred to a hatchery set at 37.0°C temperatures and RH was increased to 75% and turning of eggs was stopped in all batches. At the end of the incubation period, unhatched eggs were collected and counted.
Fatty acid profile
Gas Liquid Chromatography (GLC) method was applied for the analysis of fatty acids (FA). Fatty acid methyl esters (FAME) were prepared by transesterification (Garces and Mancha, 1993), and 1 ^l FAME was introduced to the GLC set and the resolution for each fatty acid was recorded. Standards of Fas were injected under the same temperature and pressure, and unknown fatty acids were detected by comparing obtained parameters with standards' ones. The levels of each FA in the FAME were measured by Shimadzu CR4-A Chromatopac. Thrombogenicity index and atherogenicity index (AI) and were assessed with the following formula:
Al = [(£712: 0 + 4 X C14: 0 4- C16:0)]/(£AGMl +
Haemagglutination inhibition test
All groups were vaccinated subcutaneously in the breast at 49 days with the killed AI-ND (H9N2 subtype) vaccine. Blood samples were taken on day 42 for ND antibodies and AI antibodies. Blood samples were left without anticoagulants to clot. The serum was dissociated by centrifugation at 3000 rpm for 10 min. Microtechnics of Haemagglutination inhibition (HI) test was performed according to Takatasy (1955). The Geometric mean titer (GMT) was calculated according to Brugh (1978).
Statistical analysis
Obtained data were analyzed by SPSS 16.0 (SPSS Inc., USA). Kolmogorov-Smirnov and Levene tests were applied to determine normality and homogeneity of the variances, respectively. Parametric data were presented as means ± standard deviation, and were compared between the dietary groups by one-way ANOVA and Duncan multiple comparison test (Duncan, 1995). The differences were considered statistically significant with p < 0.05.
RESULTS
Determination of minerals
The effect of Moringa seed extract and Moringa seed powder on the mean values of four minerals are shown in Table 2. Our results revealed that the iron (Fe) content of Chukar partridge eggs increased in response to the increase of dietary Moringa seed extract supplement (p > 0.05). The statistical analysis of data revealed that the
group supplemented with 1% Moringa seed powder recorded the highest Copper among different groups (p > 0.05). However, copper and magnesium levels in the Chukar partridge eggs were not significantly changed compared with the control group (p < 0.05).
Egg cholesterol, haugh unit, hatchability
Table 3 indicates the effect of Moringa seed extract and Moringa seed powder on the mean values of Haugh unit and cholesterol composition for different treatments. All treatment groups had lower levels of egg yolk cholesterol compared to the control group. Hatchability rate (Figure 1) and Haugh unit fractions were, respectively, increased and decreased in all treatments in comparison to the control group. The highest hatchability and the lowest value rates were recorded in the 1% Moringa seed extract and 0.5% Moringa seed powder supplemented groups, respectively.
Fatty acid profile
Fatty acid contents of total lipid in egg yolk in different treatments are presented in Table 4. The findings revealed that C18:1, C17:0, and C16:0 of Chukar partridge eggs increased in response to the increase of dietary Moringa seed supplementation, however, decreases were remarkable in C18:0 and C18:2.
Antibody titer against ND and AI virus
The result of the Antibody titers (Avian Influenza and Newcastle disease) of Chukar Partridge samples are presented in Figures 2 and 3. Using Moringa seed extract-1% in the diet significantly increases Newcastle disease in Japanese quail in comparison to both controls and different levels of other medicinal herb powders on 69 (Figure 2). In this study, Moringa seed powder-1% and Moringa seed extract-1% all increased the titers of Avian Influenza on the day 59 days (Figure 3).
Table 2. Mineral composition of chukar partridge eggs in different treatments (mean ± SD)
Treatment T1 T2 T3 T4 T5
Copper 0.654 ± 0.1a 0.617 ± 0.68a 0.672 ± 0.096a 0.595 ± 0.098a 0.585 ± 0.11a
Zinc 7.75 ± 0.33a 6.83 ± 0.48ab 6.56 ± 1.2b 7.06 ± 0.59ab 7.25 ± 0.42ab
Magnesium 32.13 ± 0.33a 30.58 ± 1.98a 30.09 ± 3.80a 29.04 ± 1.24a 29.99 ± 0.87a
Iron 11.99 ± 0.05b 12.29 ± 1.00b 13.10 ± 1.76ab 13.29 ± 1.45ab 14.21 ± 0.60a
a-b Means within a row sharing a common superscript are not different (p < 0.05). T] T4: 0.5%Moringa seed extract, T5: 1%Moringa seed extract : Control, T2: 0.5%Moringa seed powder, T3: 1% Moringa seed powder,
Table 3. Haugh unit and cholesterol composition of chukar partridge in different treatment
Treatment T1 T2 T3 T4 T5
Cholesterol 22.25 ± 0.95a 22.00 ± 0.81a 19.25 ± 0.50b 18 ± 0.81c 16.25 ± 0.50d
Haugh unit 79.66 ± 1.52a 77.00 ± 1.73a 79.00 ± 1a 79.33 ± 1.52a 79.00 ± 1a
a-c Means within a row sharing a common superscript are not different (p < 0.05). T1 T4: 0.5%Moringa seed extract, T5: 1%Moringa seed extract : Control, T2: 0.5%Moringa seed powder, T3: 1% Moringa seed powder,
Table 4. Fatty acid composition of total lipids in egg yolk of chukar partridge in different treatments
Treatment T1 (%) T2 (%) T3 (%) T4 (%) T5 (%)
C14:0 0a 0a 0a 0.07 ± 0.13a 0.24 ± 0.42a
C16:0 29.09 ± 3.59c 30.23 ± 0.93bc 30.91 ± 0.87bc 32.95 ± 0.48ab 34.78 ± 1.45a
C16:1 0b 0b 0b 0.17 ± 0.15b 0.71 ± 0.48a
C17:0 4.30 ± 0.9a 4.70 ± 1.15a 5.44 ± 1.64a 5.63 ± 0.84a 6.11 ± 1.81a
C18:0 10.51 ± 2.71a 8.96 ± 0.63ab 6.97 ± 0.65bc 5.29 ± 0.37c 5.16 ± 1.05c
C18:1 30.78 ± 1.65c 32.84 ± 1.82bc 37.37 ± 0.79a 35.11 ± 1.45ab 35.37 ± 0.66ab
C18:2 17.00 ± 1.21a 14.38 ± 1.32bc 15.74 ± 2.86a 14.69 ± 1.00a 14.70 ± 1.63a
C18:3 0a 0a 0a 0.03 ± 0.06a 0.09 ± 0.16a
Unknown 2.24 ± 0.15b 2.85 ± 0.33ab 3.17 ± 0.22a 2.96 ± 0.06ab 2.91 ± 0.57ab
a-c Means within a row sharing a common superscript are not different (p < 0.05). TV Control, T2: 0.5%Moringa seed powder, T3: 1%Moringa seed powder, T4: 0.5%Moringa seed extract, T5: 1%Moringa seed extract
Figure 1. Hatchability of chukar partridge in different treatments. TV Control, T2: 0.5%Moringa seed powder, T3: 1%Moringa seed powder, T4: 0.5%Moringa seed extract, T5: 1%Moringa seed extract. The a-c above the columns show significant differences between each group (p < 0.05).
A
C
8 7 6
2 5
B> 4
3 2 1 0
ab
Tl-ND T2-ND T3-ND T4-ND T5-ND Treatment
B
3.4 3.35 3.3
2 3.25 ft ,„
o 3.2
3.15
3.1 3.05
ns
Jj
Tl-ND T2-ND T3-ND T4-ND T5-ND Treatment
10
9.5
0.
5 8.5
D
7.5
ab
ab
ab
Tl-ND T2-ND T3-ND T4-ND T5-ND Treatment
Figures 2. A: Effect of different dietary herbal plants ratios on antibody titer against Avian influenza (AI) virus of chukar partridge on days 49, 59, and 69. B: Avian influenza on day 49, C: Avian influenza on day 59, D: Avian influenza on day 69.
Tj: Control, T2: 0.5% Moringa seed powder, T3: 1% Moringa seed powder, T4: 0.5% Moringa seed extract, T5: 1% Moringa seed extract. The a-c above the columns shows significant differences between each group (p < 0.05).
A
B
• sV ^f tf
V < Treatment
^^^ ^^^ ^^^ ^^
C
Treatment
Figures 3. A: Effect of different dietary herbal plants ratios on antibody titer against Newcastle disease (AI) virus of chukar partridge on days 49, 59, and 69. B: Newcastle disease on day 49, C: Newcastle disease on day 59, D: Newcastle disease on
day 69. Ti: Control, T2: 0.5% Moringa seed powder, T3: 1%Moringa seed powder, T4: 0.5% Moringa seed extract, T5: 1%Moringa seed extract. The a-c above the columns shows significant differences between each group (p < 0.05).
DISCUSSION
Herbs containing biologically active substances have been evaluated in so many studies for curative and health properties agents (Gerzilov et al., 2015) and now serve as a possible alternative nutritional supplement for optimal health, quantity, and quality of poultry products (Khan et al., 2017). M. oleifera has been reported to be rich in potassium, zinc, calcium, iron, and magnesium (Gopalakrishnan et al., 2016). Moringa powder has been also used for iron supplementation for the treatment of anemia. Moringa has been found to have more iron levels than spinach (Gopalakrishnan et al., 2016). Iron levels of Chukar partridge eggs were found to be the highest in the group with 1% Moringa seed extract (p > 0.05) and we believe that the reason is high levels of iron in M. oleifera (Portugaliza and Fernandez, 2012). It has been reported that phytogenic additives may significantly change the
levels of minerals in poultry products (Herke et al., 2016a; Herke et al., 2016b).
Declined levels of cholesterol in the groups treated with M. oleifera powder might be because of a P-sitosterol because of its structural similarity to cholesterol, so, it can decrease the intestinal absorption of cholesterol (Marrufo et al., 2013; Ahmad et al., 2018). Avian embryos are supplied by lipids of the yolk, and the membrane phospholipids of many cell types in the embryo have characteristic fatty acid profiles which are related to the special functions and particular stages of tissues differentiation (Surai et al., 2001). The findings of this article showed that C18:1, C17:0, and C16:0 of Chukar partridge eggs increased in response to the increase of dietary Moringa seed supplementation, however, decreases were remarkable in C18:0 and C18:2. It has been found that a diet supplemented by M. oleifera can reduce the levels of short-chain fatty acids, palmitic acid, cholesterol in serum and meat, and increase the levels of unsaturated
fatty acids (UFA) in chickens (Kout Elkloub et al., 2015; Mabusela et al., 2018). Antioxidant agents such as flavonoids, ascorbic acid, and phenolics compounds could be responsible for the inhibition of cholesterol synthesis and unsaturated fatty acid could be increased by M. oleifera (Speake et al., 1998; Marrufo et al., 2013). There is a need for antioxidant protection in chicken embryonic tissues as they have high amounts of highly polyunsaturated fatty acids in their lipid fraction. High levels of endogenous antioxidants are critical for embryonic tissue protection during hatching as oxidative stress (Surai et al., 2016). The increase in hatchability at higher levels of M. oleifera seed meal can be attributed to the increase of unsaturated fatty acids (Gakuya et al., 2014; Surai et al., 2016).
Using Moringa seed extract-1% in the diet significantly increases Newcastle disease in Japanese quail in comparison to both controls and different levels of other medicinal herb powders on 69 days. In this study, Moringa seed powder-1% and Moringa seed extract-1% all increased the titers of Avian Influenza on 59 days. Nonspecific defense mechanisms, and humoral and cellular immunities of the animal immune system have been stimulated and suppressed by herbal plant supplements. Nutrition is a critical determinant of immune responses. Natural products can be used as immunostimulants (Stanwell-Smith, 2001; Yassein et al., 2015). Medicinal plants having glycosides and carbohydrates are considered beneficial to immune system mechanisms by increasing body power (Yadav et al., 2014). Antimicrobial compounds (lipophilic compounds) and antioxidants (polyphenols, tannins, anthocyanins, glycosides) in M. oleifera may bind to the cytoplasmic membrane and destroy free radicals, activating antioxidant enzymes. As a result, it inhibits oxidases and therefore these elements are more available to birds (Jabaeen et al., 2008). Vitamins A, C and E as well as their provitamins existing in M. oleifera leaves are known to embay free radicals and may have immune protective effects (DanMalam et al., 2001).
CONCLUSION
Dietary supplementation with 0.5% or 1% Moringa seed powder and extract can improve cholesterol levels, hatchability, fatty acid profiles, and iron of eggs during storage, without any adverse effect on either laying performance or egg quality in chukar partridge. Therefore, it can be concluded that 1% of Moringa seed powder and extract are beneficial additives to the diets of chukar partridge.
DECLARATIONS
Acknowledgments
The authors would like to thank Mr. Ali Kameli for her help with the project during the experimental period.
Competing interests
The authors of this study declare no conflict of interest.
Authors' contribution
Habibi and Ghahtan were involved in the data collecting, statistical analysis, and drafting of the manuscript. Kohanmoo read and approved the final manuscript.
Ethical consideration
Ethical issues (Including plagiarism, consent to publish, misconduct, data fabrication and/or falsification, double publication and/or submission, and redundancy) have been checked by the authors.
REFERENCES
Abdulkarim SM, Long K, Lai OM, Muhammad SKS and Ghazali HM (2005). Some physico-chemical properties of Moringa oleifera seed oil extracted using solvent and aqueous enzymatic methods. Food Chemistry, 93, 253-263. DOI:
https://doi.org/10.1016/j.foodchem.2004.09.023
Aboonajmi M, Akram A, Nishizu T, Kondo N, Setarehdan SK and Rajabipour A (2010). An ultrasound based technique for the determination of poultry egg quality. Research in Agricultural Engineering, 56: 26-32. DOI: https://doi.org/10.17221/18/2009-RAE
Ahmad S, Khalique A, Pasha TN, Mehmood S, Hussain K, Ahmad S, Shaheen MS, Naeem M and Shafiq, M (2017) Effect of Moringa oleifera (Lam.) pods as feed additive on egg antioxidants, chemical composition and performance of commercial layers. South African Journal of Animal Science, 47: 864-874. DOI: https://doi.org/10.4314/sajas.v47i6.14
Alikwe PCN and Omotosho MS (2013). Evaluation of the proximate, chemical and phytochemical composition of Moringa oleifera leaf meal as potential food/feed stuff for man and Non ruminant livestock. Agrosearch, 13: 17-28. DOI:
https://doi.org/10.4314/agrosh.v13iL2
Association of Official Analytical Chemists (AOAC) (1990). Association of official analytical chemists. Official Methods of Analysis, 15th Ed (Washington), USA.
Ayasan T (2015). Use of Moringa oleifera in poultry and ruminant nutrition. Türk Tarim Gida Bilim ve Teknoloji Dergisi, 3: 425-429. DOI: https://doi.org/10.24925/turjaf.v3i6.425-429.327
Behnamifar A, Rahimi S, Karimi Torshizi MA, Hasanpour S, and Mohammadzade Z (2015). Effect of thyme, garlic and caraway herbal extracts on blood parameters, productivity, egg quality, hatchability and intestinal bacterial population of laying Japanese quail. Iranian Journal of Veterinary Medicine, 9: 179-187. DOI: https://doi.org/10.22059/IJVM.2015.55286
Brugh TM. (1978). A simple method for recording and analyzing
serological data. Avian Diseases, 22: 362-365. DOI: https://doi.org/10.2307/1589552
Barbanera F, Guerrini M, Hadjigerou P, Panayides P, Sokos C, Wilkinson P, Khan AA, Khan BY, Cappelli F, and Dini F (2007). Genetic insight into Mediterranean chukar (Alectoris chukar, Galliformes) populations inferred from mitochondrial DNA and RAPD markers. Genetica, 131: 287-298. DOI: https://doi.org/10.1007/s10709-006-9138-x.
Castillo LRI, Portillo LJJ, León FJ, Gutiérrez DR, Angulo EMA, Muy-Rangel MD and Heredia JB (2018). Inclusion of Moringa leaf powder (Moringa oleifera) in fodder for feeding Japanese Quail (Coturnix coturnix japonica). Brazilian Journal of Poultry Science, 20: 15-26. DOI: https://doi.org/10.1590/1806-9061-2017-0410
Caglayan T, Kirikci K and Aygun A (2014). Comparison of hatchability and some egg quality characteristics in spotted and unspotted partridge (Alectoris chukar) eggs. The Journal of Applied Poultry Research. 23 (2): 244-251. DOI: https://doi.org/10.3382/iapr.2013-00899
Compaoré WR, Nikiéma PA, Bassolé HIN, Savadogo A, Mouecoucou J, Hounhouigan DJ and Traoré SA (2011). Chemical composition and antioxidative properties of seeds of Moringa oleifera and pulps of Parkia biglobosa and Adansonia digitata commonly used in food fortification in Burkina faso. Current Research Journal of Biological Sciences, 3: 64-72. DOI: https://doi.org/10.12691/aifi-6-4-4
Dan-Malam HU, Abobakar Z ad Katsayal (2001). Pharmacognostical studies of Moringa oleifera Lam. seeds. Nigerian Journal of Natural Products and Medicine, 5: 45-49. DOI: https ://doi .org/10.4314/ninpm.v5i1.11723
Duncan D (1995). Multiple range and multiple F test. Biometrics, 11: 142. DOI: https://doi.org/10.2307/3001478
Gadzirayi CT, Masamha B, Mupangwa JF and Washaya S (2012). Performance of broiler chickens fed on mature Moringa oleifera leaf meal as a protein supplement to soyabean meal. International Journal of Poultry Science, 1: 5-10. DOI: https://doi.org/10.3923/iips.2012.5.10
Gakuya DW, Mbugua PN, Mwaniki SM, Kiama SG, Muchemi GM and Njuguna A (2014). Effect of supplementation of Moringa oleifera (LAM) leaf meal in layer chicken feed. International Journal of Poultry Science, 13: 379-84. DOI:
https://doi.org/10.3923/ijps.2014.379.384
Gerzilov V, Nikolov A, Petrov P, Bozakova N, Penchev G and Bochukov A (2015). Effect of a dietary herbal mixture supplement on the growth performance, egg production and health status in chickens. Journal of Central European Agriculture, 16: 10-27. DOI: https://doi.org/10.5513/JCEA01/16.2.1580
Habibi H, Ghahtan N, and Brooks DM (2019). Effect of sex ratio, storage time and temperature on hatching rate, fertility and embryonic mortality in Chukar partridge (Alectoris chukar). Animal Reproduction Science, 203: 68-74. DOI: https://doi.org/10.1016/i.anireprosci.2019.02.009
Herke R, Gálik B, Bíro D, Rolinec M, Jurácek M, Arpásová H and Hanusovsky O (2016a). The effect of essential plant oils on mineral composition of egg mass and blood parameters of laying hens. Journal of Central European Agriculture, 17, 1150-67. DOI: https://doi.org/10.5513/JCEA01/17.4.1824
Herke R, Gálik B, Bíro D, Rolinec M, Jurácek M, Arpásová H and Wilkanowska A (2016b). The effect of a phytogenic additive on nutritional composition of turkey meat. Journal of Central European Agriculture, 17: 25-39. DOI:
https://doi.org/10.5513/JCEA01/17.L1664
Alabi OJ, Malik AD, Ng'ambi JW, Obaje P and Ojo BK (2016). Effect of Aqueous Moringa Oleifera (Lam) Leaf Extracts on Growth Performance and Carcass Characteristics of Hubbard Broiler Chicken. Brazilian Journal of Poultry Science, 19(2):273-280. DOI: https://doi.org/10.1590/1806-9061-2016-0373
Food and Agriculture Organization (FAO) (2009). How to Feed the World in 2050.
http://www.fao.org/fileadmin/templates/wsfs/docs/expert paper/Ho w_to_Feed_the_World_in_2050.pdf
Garces R and Mancha M (1993). One-Step lipid extraction and fatty acid methyl esters preparation from fresh plant tissues. Analytical Biochemistry, 211: 139-143. DOI:
https://doi.org/10.1006/abio.1993.1244
Gopalakrishnan L, Doriya K and Kumar DS (2016). Moringa Oleifera: A review on nutritive importance and its medicinal application. Food Science and Human Wellness, 2: 49-56. DOI: https://doi.org/10.1016/j .fshw.2016.04.001
Khan I, Zaneb H, Masood S, Yousaf MS, Rehman HF and Rehman H (2017). Effect of Moringa oleifera leaf powder supplementation on growth performance and intestinal morphology in broiler chickens. Journal of Animal Physiology Animal Nutrition, 101: 114-21. DOI: https://doi.org/10.1111/ipn.12634
Kout Elkloub ME, Moustafa R, Shata FH, Mousa MA, Hanan AH, Alghonimy and Youssef SF (2015). Effect of Using Moringa Oleifera leaf meal on performance of Japanese Quail. Egyptian Poultry Science Journal, 35: 1095-108. DOI: https://doi.org/10.21608/EJNF.2017.104115
Marrufo T, Nazzaro F, Mancini E, Fratianni F, Coppola R, De Martino L, Agostinho AB and De Feo, V. (2013). Chemical composition and biological activity of the essential oil from leaves of Moringa oleifera Lam. cultivated in Mozambique. Molecules, 18: 1098911000. DOI: https://doi.org/10.3390/molecules180910989
Mabusela SP, Nkukwana TT, Mokoma M and Muchenje V (2018). Layer performance, fatty acid profile and the quality of eggs from hens supplemented with Moringa oleifera whole seed meal. South African Journal Animal Science, 48: 234-43. DOI: https://doi.org/10.4314/sajas.v48i2.4
Marrufo T, Nazzaro F, Mancini E, Fratianni F, Coppola R, De Martino L, Agostinho AB and De Feo V (2013). Chemical composition and biological activity of the essential oil from leaves of Moringa oleifera Lam. cultivated in Mozambique. Molecules, 18: 1098911000. DOI: https://doi.org/10.3390/molecules180910989
Nkukwana T, Muchenje V, Pieterse E, Masika P, Mabusela T, Hoffman L and Dzama K (2014). Effect of Moringa oleifera leaf meal on growth performance, apparent digestibility, digestive organ size and carcass yield in broiler chickens. Livestock Science, 161: 139-146. DOI: https://doi.org/10.1016/Uivsci.2014.0L001
Portugaliza HP and Fernandez TJ (2012). Growth performance of cobb broilers given varying concentrations of malunggay (Moringa oleifera Lam.) aqueous leaf extract. Online Journal of Animal and Feed Research, 2: 465-9. Available at: https://www.cabdirect.org/cabdirect/abstract/20133038898
Pourghanbari GH, Nili H, Habibi H, Morovati M, Salehi E, and Sadoughifar R (2016). Response of Chukar partridge performance and blood parameters to different dietary crude protein. Advances in Bioresearch, 7: 162-169. DOI:
https://doi.org/10.13140/RG.2.1.3282.0084
Rehault-Godbert S, Guyot N, Nys Y (2019). The Golden Egg: Nutritional Value, Bioactivities, and Emerging Benefits for Human Health. Nutrients, 11 (684): 1-26. DOI: https://doi.org/10.3390/nu11030684
Speake BK, Murray AMB, Noble CR (1998). Transport and transformations of yolk lipids during development of the avian embryo. Progress in Lipid Research, 37: 1-32. DOI: https://doi .org/10.1016/S0163 -7827(97)00012-X
Stanwell-Smith R. 2001. Hygiene and the immune system. American Society for Clinical Nutrition, 43: 61-64. DOI: https://doi.org/10.1053/iinf.2001.0859
Surai PF, Bortolotti GR, Fidgett AL, Blount JD, Speake BK (2001). Effects of piscivory on the fatty acid profiles and antioxidants of avian yolk: Studies on eggs of the gannet, skua, pelican and
cormorant. The Zoological Society of London, 255: 305-312. DOI: https://doi.org/10.1017/S0952836901001406
Surai P F, Fisinin VI and Karadas F (2016). Antioxidant systems in chick embryo development. Part1. Vitamin E, carotenoids and selenium. Animal Nutrition, 2: 1-11. DOI: https://doi.org/10.1016/j .aninu.2016.01.001
Takatsy G (1955). The use of spiral loops in serological and virological micromethods. Acta Microbiologica et Immunologica Hungarica 3:191-202. https://doi.org/10.1556/AMicr.50.2003A5
Yassein DMM, Abdallah EA, Ismail II, and Faddle AA (2015). Effect of dietary supplementation of pomegranate poeel powder and butylated hydroxy toluene on some productive, physiological and immunological parameters of Japanese quail. Egyptian journal of Animal Production, 52: 105-113. DOI:
https://doi.org/10.21608/ejap.2015.170899
Windisch W, Schedle K, Plitzner C and Kroismayr A (2008). Use of phytogenic products as feed additives for swine and poultry. Animal Science Journal, 86: 140-148. DOI: https://doi.org/10.2527/jas.2007-0459
Worku A (2016). Moringa oleifera as a Potential Feed for Livestock and Aquaculture Industry. African Journal of Agricultural Science and Technology, 4 (4): 666-676. https://docplayer.net/53686830-Moringa-oleifera-as-a-potential-feed-for-livestock-and-aquaculture-industry.html
Yadav M, Jhunjhunwala, Phung TP, Lupardus P, Tanguay J, Bumbaca S, Franci C, Cheung TK, Fritsche J, Weinschenk T, Modrusan Z, Mellman I, Lill JR and Delamarre L (2014). Predicting immunogenic tumour mutations by combining mass spectrometry and exome sequencing. Nature, 515: 572-576. DOI: https://doi.org/10.1038/nature14001
