1. Introduction
The global demand for an adequate and reliable supply of milk is predicted to increase by 35% by 2030, owing primarily to increased demand in Asia [1,2]. As a result, in recent decades, various countries—including Saudi Arabia—have encouraged the expansion and development of dairy production in order to fulfil increasing consumer demand [3,4]. However, various socioeconomic and environmental issues confront milk production strategies, including the risks linked with the use of growth promoters and antibiotics in livestock [5,6]. Such difficulties have inspired a worldwide search for natural feed supplements that can improve cow milk yields [7,8,9].
Condensed tannins, for example, have demonstrated a range of antimicrobial properties in ruminants, as well as a great potential to affect the rumen environment, with the specific consequence of enhancing milk efficiency and nitrogen use in lactating dairy cows [10]. Moreover, some earlier reported studies highlighted the capacity of condensed tannin to mitigate methane emissions from cattle [11,12]. Essential oils, such as cinnamaldehyde with its potent antimicrobial activity, have been shown to favorably modify dairy cow performance and rumen fermentation [13,14]. Additionally, various beneficial effects have been reported for plant-based bioactive compounds such as curcumin, such as boosting immune status and protecting against infectious disease in dairy cows [15] and dairy sheep [16]. Additionally, capsaicin—an active phenolic compound in Capsicum spp.—has been established to be an antibacterial agent that improves rumen fermentation [17], a regulator of glucose homeostasis through an insulin secretion mechanism, and a stimulant of digestive enzyme secretion [18]. Furthermore, other bioactive plant secondary components such as piperine have been reported to improve rumen fermentation and boost animal production efficiency and health [19,20].
Based on the information provided, it was hypothesized that feeding a blend of encapsulated cinnamaldehyde, condensed tannins, and plant-based bioactive compounds could improve lactation performance and dairy cow health. Hence, this study evaluated the effects of using Actifor® Pro as a functional feed supplement (FFS) containing the above-mentioned ingredients on the milk production efficiency and composition of multiparous Holstein cows during mid-lactation. Additionally, blood biochemical indicators of liver and kidney function as well as the cows’ electrolyte balance and protein profiles were also studied to evaluate the safety of this product.
2. Materials and Methods
2.1. Animals, Diets, Investigational Design, and Housing
The experiment was conducted between September and November 2020 at the National Agricultural Development Company (NADEC) in Haradh, Kingdom of Saudi Arabia. All procedures and treatments in the current study were approved by the Research Ethics Committee at the King Faisal University. The present study included sixty Holstein multiparous (mean ± SD: 3.71 ± 0.8 lactations) dairy cows. The experimental animals were on average (mean ± SD) 116.1 ± 17.1 days in milk, 606 ± 9.3 kg body weight, and producing 45.73 ± 6.7 kg/d milk yield. Cows were grouped by parity and daily milk yield and then randomly allocated to the experimental groups in a completely randomized block design. Animals were randomly assigned to the control group (CON: n = 30), which received a total mixed ration (TMR) diet; or the FFS (n = 30) treatment group, which received TMR fortified with a mixture of functional feed supplements at a rate of 35 g/cow/day. The amount of supplement (Actifor® Pro, Delacon company, Engerwitzdorf, Austria) used in this study followed the recommendation of the production company. Actifor® Pro is a patented formulation that contains encapsulated cinnamaldehyde, curcumin, condensed tannins, capsaicin, and piperine. The experiment was continued for 70 days in total.
Daily feed intake was measured throughout the experiment by subtracting the offered feed from the uneaten feed. All cows were subjected to a 14-day ration adaptation period before being put on their assigned diet for a 70-day supplementation period (experimental period). All cows were kept in tie-stalls in a mechanically ventilated barn, with free access to TMR and freshwater. At the start and during the trial, the dairy cows’ health status was checked and recorded. The TMR consisted of concentrates and forages at a ratio of 53.6:46.4, respectively (DM basis). A description of the formulation and proximate composition of the TMR is presented in Table 1. Throughout the experiment, TMR samples were taken fortnightly and kept at −20 °C until further analysis.
2.2. Ration Chemical Analysis
By the end of the study, composite samples of the TMR were dried at 60 °C for 48 h in a forced-air oven and ground in a Wiley mill (Arthur H. Thomas, Philadelphia, PA, USA) to pass through a 1 mm screen. The chemical composition of the TMR was determined using AOAC [21] procedures. The content of dry matter (procedure 934.01) and organic matter (procedure 942.05) was estimated in all samples. Crude protein (N × 6.25) content estimation was performed using the Kjeldahl method (procedure 990.03). Ether extract (procedure 2003.06) content was estimated using petroleum ether in a Soxhlet extractor (Sigma-Aldrich, St. Louis, MO, USA). Acid detergent fiber (ADF) and neutral detergent fiber (NDF) levels were measured following the methods of the AOAC [21] (Method 973.18) and Van Soest, et al. [22], respectively. All chemical analyses were calculated on a dry-matter basis (DM).
2.3. Milk Yield and Composition
Milk was sampled on days 1, 14, 28, 42, 56, and 70. Cows were machine-milked daily at 4.00, 12.00, and 20.00 h, and composite samples (20 g/kg milk production) were taken at each milking time to determine the milk composition. An equal proportion from each milking was composited for each cow and sent immediately to NADEC laboratories for analysis. Milk samples were evaluated for protein, lactose, fat, ash, and total solids using mid-infrared (FTIR) spectrophotometry (MilkoScan™ FT1, SCANCO, San José, Costa Rica). The total somatic cell count in the milk samples was measured using a FossomaticTM device 7 (FOSS, Hilleroed, Denmark).
The data for the energy-corrected milk (ECM), fat-corrected milk (3.5% FCM), gross energy content of the milk, and the milk energy output were calculated as follows [24,25,26]:
ECM (kg/day) = daily milk yield (kg) × [20.7 + protein (g/kg) × 24.2 + fat (g/kg) × 38.3 + lactose (g/kg) × 16.54]/3140 (1)
3.5% FCM = (fat yield (kg) ×16.216) + (milk yield (kg) × 0.4324)(2)
Milk energy content (MJ/kg) = 4.184 × [(protein (g/kg) × 24.13 + fat (g/kg) × 41.63 + lactose (g/kg) × 21.60 − 117.2)/10000] × 2.204(3)
Milk energy output (MJ/d) = daily milk yield (kg/d) × milk energy content (MJ/kg).(4)
2.4. Serum Biochemistry
Blood samples (10 cows per treatment) were collected before the morning feed on days 1, 14, 28, 42, 56, and 70 of the experiment. The blood was taken from the tail vein (vena coccygeal) into a sterile tube without anticoagulants. After clotting, serum samples were separated by centrifuging at 2000× g and 4 °C for 10 min. Then, the resulting serum was transferred to 2 mL labeled plastic tubes and stored at −80 °C until analysis. Serum biochemical parameters were analyzed using a fully automatic biochemical analyzer (VetScan VS2, Abaxis, Inc., Union City, CA, USA). Serum biochemical measurements were conducted to determine the serum levels of total protein (TP), albumin (ALB), gamma glutamyl transferase (GGT), creatine kinase (CK), alkaline phosphatase (ALP), blood urea nitrogen (BUN), alanine aminotransferase (AST), magnesium (Mg), inorganic phosphorus (PHOS), and calcium (Ca). The globulin content (GLO) was computed by subtracting the ALB from the TP.
2.5. Statistical Analysis
All quantitative data (dry-matter intake, milk yield, and composition and blood profile) were analyzed with the period (week) as a repeated measure using the PROC MIXED method of SAS [27] and individual cows as the trial unit. The statistical model included the effects of supplementation, period, and their interaction (supplementation × period). To achieve normality, somatic cell count values were log-transformed (base-10 log) before analysis. Continuous parameter values at the end of the 14-day adaptation period were included as covariates in each of the respective data analyses. The significance level was set at a p-value of less than 0.05 (p ≤ 0.05), and a tendency was accepted when p ≤ 0.10 and >0.05. In case of the existence of significant effects, means were compared using the least squares means probability of difference.
3. Results
3.1. Lactation Performance, Feed Intake, and Feed Efficiency
Figure 1 illustrates the daily milk yield of lactating Friesian cows fed with the experimental diets (control vs. FFS) over ten weeks. Despite the higher values observed in the daily milk yield in the FFS group compared to the control group during all weeks of the experiment (Figure 1), the treatment × period interaction was not significant. Daily milk production (p = 0.01) and solids-not-fat yield (p = 0.05) were significantly higher in cows that had received FFS compared to the control group (Table 2). In addition, 3.5% FCM, ECM, lactose and protein yields, and milk energy output tended to be greater (p ≤ 0.10) in dairy cows that consumed FFS during the experimental period. No interaction effect between the treatment and period was noted for milk yield and composition (Table 2).
A significant treatment × period interaction was identified (p ≤ 0.02) with respect to the milk efficiency and somatic cell count. Cows receiving FFS had a lower (p = 0.049) somatic cell count on weeks 6–8 of the experiment compared with those on the control diet (Figure 2A). In addition, the dry-matter intake tended to be higher (p = 0.064) in animals that consumed FFS during weeks 0–2 and 2–4 of the trial period (Figure 2B). FFS supplementation significantly increased the feed efficiency, expressed as ECM /DM intake (p = 0.046), and milk yield/DM intake (p = 0.011) on weeks 4–6 and 6–8 of the experiment (Figure 2C,D).
3.2. Blood Measurements
Most serum biochemical parameters were not changed (p ≥ 0.114) between the FFS and control cows, including TP, GLO, BUN, CK, ALP, AST, GGT, Ca, PHOS, and Mg (Table 3). However, a greater concentration of serum ALB (p = 0.007) was detected in cows supplemented with FFS. There were no significant treatment × time interactions for the serum biochemical parameters. Interactions between treatment and time with respect to the concentrations of serum hepatic enzymes (ALP, AST, and GGT) were not detected (p > 0.05; Figure 3). In addition, the serum protein parameters BUN and CK were not significantly affected by interactions between treatment and time (Figure 4 and Figure 5). The treatment × day interaction was also not significant for the major serum minerals measured (Figure 6).
4. Discussion
In the current experiment, lactating Holstein cows fed FFS-fortified diets had significantly greater milk production compared to those fed with control diets. The FFS-associated boost in milk production could be highly related to the biological activities of its ingredients. In this regard, the addition of 100 mg curcumin/kg diet for 15 days has a beneficial impact on the milk yield of lactating ewes [16]. Additionally, an obvious increase in milk production was recorded for cows fed diets fortified with an herb mixture comprising Curcuma longa [28]. Additionally, earlier studies found that adding an essential oil blend containing cinnamaldehyde to lactating cows enhanced milk yield [29,30]. The favorable impact of cinnamaldehyde in improving the rumen fermentation efficiency could be responsible for optimizing milk yield [31,32]. Additionally, in a recent study by Abulaiti et al. [33], capsaicin supplementation in Chinese Holstein dairy cows maintained milk production and composition. Furthermore, the increasing tendency in the dry-matter intake of lactating Holstein cows fed with FFS-fortified diets could also contribute to the enhancement of milk production [34].
Of note, in the current study, changes in not only the milk yield were observed by the addition of FFS to the lactating Friesian cows’ feed, but also in the milk’s composition. A significant increase in the solids-not-fat yield and an increasing tendency in the lactose and protein yields were recorded in Holstein cows fed FFS-fortified feeds compared with cows in the control group. However, a significant decrease in the somatic cell count was recorded in the milk of cows of the FFS groups compared to control group. Similarly, supplementing lactating dairy cows with a blend of cinnamaldehyde, thymol, and orange peel increased the protein content of their milk [35]. Additionally, supplementing dairy ewes with a 100 mg curcumin/kg diet decreased the somatic cell count in their milk [16]. The anti-inflammatory and antioxidant actions of curcumin could act both in the mammary gland and systemically, controlling inflammatory responses and reducing inflammatory cells in milk [36,37]. Importantly, the presence of piperine in the formulation could potentiate the biological activities of curcumin [38]. Additionally, when capsaicinoids are injected postruminally, they trigger significant changes in the physiological responses linked to inflammation and milk yield in lactating cows [39]. Moreover, several research works have suggested that low somatic cell count values are correlated with increased milk production [40,41]—a finding observed in the current investigation.
Feed additives must be safe for the health and wellbeing of a cow to support her milk production and milk quality [42]. In the present study, several blood biochemical indicators were estimated to determine the safety of FFS as a feed supplement for dairy cows. Measurements of the serum activities of the hepatic enzymes AST, GGT, and ALP are considered as a reliable indicator of liver function in dairy cows [43,44,45]. Additionally, CK has been used as a screening parameter in the diagnosis of endometritis muscular damage or hypocalcemia in dairy cows [46]. The present data on serum AST, GGT, ALP, and CK show that no significant alterations exist in Holstein cows fed FFS-fortified diets compared to those fed control diets. Hence, the provision of FFS efficiently enhanced the cows’ performance without affecting their liver function or muscle condition.
Serum levels of TP, GLO, and BUN have been extensively used in dairy animals to determine the amount of protein required for milk production [47]. The normal ranges that have been recorded demonstrate the optimal conditions for milk production. BUN levels may increase during water deprivation [48], thirst, diarrhea, urinary diseases [49], pregnancy toxemia [50], and acidosis [51], none of which were the case in the current study. Cows fed FFS showed higher serum ALB levels (p = 0.007) than those fed with the CON diet. This could be because cows fed with FFA have a tendency to increase their dry-matter intake, as observed in the current experiment—or, it could be due to an increase in the amino acid availability of cows fed with FFA. Waghorn, et al. [52] observed a significant increase in post-ruminal amino acid flux and intestinal amino acid availability as a result of feeding with condensed tannins.
Major blood mineral concentrations provide an indication of the health status of dairy animals. The role of macro minerals such as Ca, Phos, and Mg are more important than other minerals, and their balance in the body improves animal health, reproduction, and production performance [47]. Hypomagnesaemia or hypocalcemia in high-producing dairy cows increases the risk of some disorders such as milk fever [53]. Hypocalcemia reduces milk production and increases diarrhea and vagus indigestion due to the inhibition of feed intake [54]. Herein, serum mineral concentrations within the normal range reflect that animal diets contain an adequate amount of minerals [47].
5. Conclusions
Overall, the results showed that the dietary application of Actifor® Pro as a functional supplement mixture including encapsulated cinnamaldehyde, condensed tannins, capsaicin, piperine, and curcumin to Holstein lactating cows at 35 g/cow/day enhanced milk production and feed efficiency. Moreover, the functional supplement mixture increased blood albumin levels and had no negative effect on other blood measurements or milk composition. Further studies are recommended to evaluate the effects of functional supplement mixtures on the physiological and metabolic aspects of high-producing cows during the transition period.
Conceptualization, S.B.A., A.M.A., Y.M.A. and A.T.M.; methodology, S.B.A., A.M.A., Y.M.A. and A.T.M.; software, S.B.A. and A.T.M.; formal analysis, A.A.A.-S.; investigation, S.B.A., A.M.A. and Y.M.A.; resources, S.B.A., A.M.A. and Y.M.A.; data curation, S.B.A., A.M.A., Y.M.A. and A.T.M.; writing—original draft preparation, A.A.A.-S.; writing—review and editing, S.B.A.; supervision, S.B.A.; funding acquisition, S.B.A. All authors have read and agreed to the published version of the manuscript.
Not applicable.
The datasets used along with this research are available from the corresponding author upon reasonable request.
The authors would like to acknowledge the Deanship of Scientific Research, Vice Presidency for Graduate Studies and Scientific Research, King Faisal University, Saudi Arabia.
The authors declare no conflict of interest.
Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Enlarge this image.Figure 1. Milk yield of the lactating Friesian cows assigned to receive 35 g/cow/d of a functional supplement mixture (FFS group; n = 30), or not (Control group; n = 30), for 10 weeks. The supplement contained condensed tannins, essential oils, and pungent compounds (Actifor Pro; Delacon Biotechnik GmbH; Steyregg, Austria). The values shown are the means ± SE.
Enlarge this image.Figure 2. Somatic cell count (A), dry-matter intake (B), and feed efficiency (C,D) of lactating Friesian cows assigned to receive 35 g/cow/d of functional supplement mixture (FFS group; n = 30), or not (Control group; n = 30), for 10 weeks. The supplement contained condensed tannins, essential oils, and pungent compounds (Actifor Pro; Delacon Biotechnik GmbH; Steyregg, Austria). The values shown are the means ± SE. Treatment comparison within weeks; * p ≤ 0.05.
Enlarge this image.Figure 3. Changes in the activity of alanine aminotransferase (AST; (A)), alkaline phosphatase (ALP; (B)), and gamma glutamyl transferase (GGT; (C)) in the blood serum of lactating Friesian cows assigned to receive 35 g/cow/d of a functional supplement mixture (FFS group; n = 10), or not (Control group; n = 10), for 70 days.
Enlarge this image.Figure 4. Changes in the concentrations of total protein (TP; (A)), albumin (B), and globulin (GLOB; (C)) in the blood serum of lactating Friesian cows assigned to receive 35 g/cow/d of a functional supplement mixture (FFS group; n = 10), or not (Control group; n = 10), for 70 days.
Enlarge this image.Figure 5. Changes in the blood urea nitrogen (BUN; (A)) concentration and creatine kinase (CK; (B)) activity of lactating Friesian cows assigned to receive 35 g/cow/d of a functional supplement mixture (FFS group; n = 10), or not (Control group; n = 10), for 70 days.
Enlarge this image.Figure 6. Changes in the concentrations of Calcium (Ca; (A)), Phosphorus (PHOS; (B)), and Magnesium (Mg; (C)) in the blood serum of lactating Friesian cows assigned to receive 35 g/cow/d of a functional supplement mixture (FFS group; n = 10), or not (Control group; n = 10), for 70 days.
Ingredients and chemical composition (on a DM basis) of the basal diet 1 fed to the lactating dairy cows during the experimental period.
| Ingredients | [g/kg Feed] | Nutrient Levels | [g/kg Feed] |
|---|---|---|---|
| Corn grain | 295.6 | Chemical analysis 3 | |
| Wheat bran | 17.7 | Organic matter | 921.6 |
| Soybean meal | 31.9 | Crude protein (N × 6.25) | 158.6 |
| Corn silage | 49.6 | Ether extract | 40.4 |
| Uncorticated cottonseed meal | 85.0 | Neutral detergent fiber | 304.0 |
| Alfalfa hay | 343.4 | Acid detergent fiber | 194.4 |
| Wheat straw | 70.8 | NFC 4 | 418.6 |
| Limestone | 12.3 | Calculated values 5 | |
| Sodium bicarbonate | 7.3 | Digestible crude protein | 669.1 |
| Canola meal oil-free | 70.7 | Metabolizable energy, Mcal/kg | 2.19 |
| Palm oil hydrogenated | 7.8 | Net energy for lactation, Mcal/kg | 1.38 |
| Calcium salt palm oil | 5.9 | NDICP | 18.4 |
| Premix 2 | 1.2 | ADICP | 12.4 |
| Distiller yeast | 0.4 | ||
| Mycotoxin binder | 0.4 |
Note: NDICP, neutral detergent insoluble protein; ADICP, acid detergent insoluble protein. 1 The basal diet based on a concentrate feed mixture and forages at a ratio of 53.6:46.4, respectively. 2 Premix contents per kg: 141 g Ca, 87 g P, 45 g Mg, 14 g S, 120 g Na, 6 g K, 944 mg Fe, 7613 mg Zn, 484 mg Cu, 748 mg Mn, 58 mg I, 81 mg Co, 13 mg Se, 248 000 IU vitamin A, 74 000 IU vitamin D3, 1656 IU vitamin E. 3 According to the AOAC [
Intake, milk yield, and composition of lactating Friesian cows fed a basal diet supplemented with a functional feed supplement mixture.
| Treatment | SEM | p-Value | ||||
|---|---|---|---|---|---|---|
| Control | FFS | Treatment | Period | Treatment × Period | ||
| Number of cows | 30 | 30 | - | - | - | - |
| Production (kg/d) | ||||||
| Milk | 45.01 | 46.59 | 0.320 | 0.010 | <0.001 | 0.918 |
| Fat corrected milk 3.5% | 34.77 | 36.43 | 0.743 | 0.100 | <0.001 | 0.206 |
| Energy corrected milk (ECM) | 34.74 | 36.15 | 0.436 | 0.093 | 0.001 | 0.329 |
| Total solids | 4.98 | 5.04 | 0.057 | 0.512 | <0.001 | 0.564 |
| Solids-not-fat | 4.03 | 4.14 | 0.030 | 0.050 | 0.091 | 0.946 |
| Fat | 0.93 | 0.99 | 0.029 | 0.285 | 0.001 | 0.184 |
| Protein | 1.50 | 1.55 | 0.011 | 0.051 | 0.030 | 0.943 |
| Lactose | 2.12 | 2.25 | 0.016 | 0.051 | 0.078 | 0.948 |
| Ash | 0.38 | 0.36 | 0.016 | 0.412 | 0.053 | 0.626 |
| Milk energy output (MJ/d) | 107.86 | 112.21 | 1.367 | 0.098 | 0.001 | 0.333 |
| Milk composition (g/kg) | ||||||
| Total solids | 110.62 | 108.15 | 0.897 | 0.169 | 0.099 | 0.525 |
| Solids-not-fat | 89.48 | 87.46 | 0.311 | 0.349 | <0.001 | 0.860 |
| Fat | 21.14 | 20.69 | 0.584 | 0.692 | 0.032 | 0.225 |
| Protein | 33.42 | 33.22 | 0.123 | 0.348 | <0.001 | 0.869 |
| Lactose | 48.70 | 48.42 | 0.165 | 0.356 | <0.001 | 0.859 |
| Ash | 8.42 | 7.60 | 0.317 | 0.200 | 0.206 | 0.715 |
| Somatic cell count (×105) | 3.68 | 2.85 | 0.55 | 0.452 | 0.395 | 0.049 |
| Milk energy content (MJ/kg) | 2.40 | 2.41 | 0.024 | 0.872 | 0.136 | 0.366 |
| Dry matter intake (kg/d) | 25.40 | 25.76 | 0.733 | 0.536 | 0.279 | 0.064 |
| Feed efficiency | ||||||
| Kg milk/kg DM intake | 1.78 | 1.81 | 0.014 | 0.187 | <0.001 | 0.011 |
| Kg ECM/kg DM intake | 1.37 | 1.40 | 0.018 | 0.361 | <0.001 | 0.046 |
SEM: standard error of the mean. The basal diet based on 536 g of concentrates feed mixture/kg DM and 464 g forages/kg DM with no additive (Control group), or with the addition of 35 g/cow/d of functional supplement mixture (FFS group).
Blood biochemistry of lactating Friesian cows fed a basal diet supplemented with a functional feed supplement mixture.
| Treatment | SEM | p-Value | ||||
|---|---|---|---|---|---|---|
| Control | FFS | Treatment | Period | Treatment × Period | ||
| Total protein, g/dL | 8.05 | 8.21 | 0.060 | 0.154 | 0.004 | 0.578 |
| Albumin, g/dL | 3.65 | 3.80 | 0.024 | 0.007 | 0.039 | 0.599 |
| Globulin, g/dL | 4.41 | 4.42 | 0.053 | 0.813 | 0.081 | 0.803 |
| Urea nitrogen, mg/dL | 15.35 | 14.93 | 0.255 | 0.424 | 0.001 | 0.927 |
| Creatine kinase, U/L | 221.23 | 273.23 | 26.123 | 0.326 | 0.659 | 0.988 |
| Alkaline phosphatase, U/L | 40.06 | 38.66 | 1.158 | 0.628 | 0.568 | 0.956 |
| Alanine aminotransferase, U/L | 70.55 | 74.66 | 1.515 | 0.114 | <0.001 | 0.566 |
| Gamma glutamyl transferase, U/L | 27.68 | 29.26 | 0.919 | 0.221 | 0.774 | 0.999 |
| Calcium, mg/dL | 9.42 | 9.34 | 0.042 | 0.417 | 0.530 | 0.928 |
| Phosphorus, mg/dL | 7.06 | 7.13 | 0.109 | 0.642 | 0.030 | 0.656 |
| Magnesium, mg/dL | 2.31 | 2.37 | 0.019 | 0.148 | 0.009 | 0.959 |
SEM: standard error of the mean. The basal diet based on 536 g of concentrates feed mixture/kg DM and 464 g forages/kg DM with no additive (Control group) or with the addition of 35 g/cow/d of a functional supplement mixture (FFS group).
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; Al-Sagheer, Adham A 3
1 Department of Animal and Fish Production, King Faisal University, P.O. Box 402, Al-Ahsa 31982, Saudi Arabia;
2 Department of Animal and Fish Production, King Faisal University, P.O. Box 402, Al-Ahsa 31982, Saudi Arabia;
3 Department of Animal Production, Faculty of Agriculture, Zagazig University, Zagazig 44511, Egypt