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. 2023 May 8;102(8):102775. doi: 10.1016/j.psj.2023.102775

Effect of water supplementation of Magic oil at different growing periods on growth performance, carcass traits, blood biochemistry, and ileal histomorphology of broiler chickens

Elsayed OS Hussein 1, Gamaleldin M Suliman 1, Abdullah N Al-Owaimer 1, Hani H Al-Baadani 1, Maged A Al-Garadi 1, Hani A Ba-Awadh 1, Mohammed M Qaid 1, Ayman A Swelum 1,1
PMCID: PMC10242640  PMID: 37269792

Abstract

Natural antibiotic substitutes have recently been used as growth promoters and to combat pathogens. Therefore, this study aimed to assess the effects of adding Magic oil (nano-emulsified plant oil) at different growing periods on growth performance, histomorphology of the ileum, carcass traits, and blood biochemistry of broiler chickens. A total of 432-day-old Ross 308 chicks were randomly assigned to 1 of 6 water supplementation treatment groups based on growing periods, with 4 groups of Magic oil programs compared to probiotic (Albovit) as a positive control and nonsupplemented group as a negative control, with 9 replicates each with 8 birds (4♂ and 4♀). The periods of adding Magic oil Magic oil were 35, 20, 23, and 19 d for T1, T2, T3, and T4, respectively. Birds’ performance was evaluated during 0 to 4, 4 to 14, 21 to 30, 30 to 35, and overall days old. Carcass parameters, blood chemistry, and ileal histomorphology were examined on d 35. The findings showed that birds in the T4 group of the Magic oil supplementation program (from 1 to 4 and 21 to 35 d of age) consumed 1.82% and 4.20% more food, gained 3.08% and 6.21% more, and converted feed to meat 1.39% and 2.07% more than Albovit and negative control, respectively, during the experiment (1–35). Magic oil particularly T1 (Magic oil is supplemented throughout the growing period) and T4 programs improved intestinal histology compared to the negative control. There were no changes (P > 0.05) between treatments in carcass parameters and blood biochemistry. In conclusion, water supplementation with Magic oil for broilers improves intestinal morphometrics and growth performance similar to or better than probiotic, especially during brooding and overall periods. Further studies are needed to evaluate the effect of adding both nano-emulsified plant oil and probiotics on different parameters.

Key words: broiler chicken, growth performance, ileal histomorphology, nano-emulsified plant oil, probiotic

INTRODUCTION

At the government and corporate levels in developing countries, there is growing interest in increasing poultry production in sufficient quantities to achieve self-sufficiency in poultry meat (Attia et al., 2022). Antibiotic resistance is a growing threat that is caused in part by incorrect or excessive antibiotic use in poultry and farm animal production (Haulisah et al., 2021; Pan et al., 2022). Consequently, the development of new types of feed additives that are resistant to drug resistance, nontoxic, and nonresidue, and the search for antibiotic alternatives to improve the performance of poultry and other livestock in the postantibiotic era are inevitable trends for future development (Chandra et al., 2021; Ruan et al., 2021). Feed additives do not only prevent or reduce a problem but also provide significant benefits to users in a variety of ways. And, while no one would suggest using a wide range of additives, prudent use of the most appropriate products pays off, and an important new consideration for feed additives is zoning additives specific to different stages of growth and development (El-Saadony et al., 2022).

Recently, the world has been striving to use antibiotic alternatives, including essential oils (EOs) and their nano-emulsions (Sharma et al., 2023), probiotics (Abd El‐Hack et al., 2020), amino acids (Gelinski et al., 2023), herbal extracts (Al-Garadi et al., 2022; Al-Garadi et al., 2023a), and green nanoparticles (Abd El-Ghany et al., 2021) to improve birds’ performance, control pathogens and avoid antibiotics resistance and misuse risks.

Nanotechnology applications in food and animal nutrition research could assist researchers in obtaining precise information data about the location of nutrients or bioactive feed constituents in cellular and tissue portions (Book chapter; Sounkaria et al., 2023). Nanotechnology has appeared as a technological breakthrough for the poultry and agri-feed industries (Prasad et al., 2022). This helps to boost global feed production while also improving feed quality, safety, and nutritional value (King et al., 2018; Nagarajan et al., 2021; Book chapter).

The use of nano-emulsions, microemulsions, polymeric nanoparticles, liposomes, polymeric micelles, and colloidal submicrometric lipid nanocarriers is a promising pharmaceutics tool (Frank et al., 2015; Mishra et al., 2018; Musielak et al., 2022). Nano-emulsions are a type of actively stable emulsion in which surfactant molecules use surface tension to stabilize the oil and water phases (Zhu et al., 2019; Zhang et al., 2021). They are used in the feed, pharmaceutical, medical applications, and cosmetic industries to coat, encapsulate and poorly disperse water-soluble bioactive ingredients. Many advantages make nano-emulsions with low turbidity and high stability rates attractive delivery systems (Abd El-Hack et al., 2021).

As powerful natural antimicrobial agents in food, nano emulsions can help EOs to be used in dietary products by increasing their resonance in the areas where microbes live, thereby enhancing their antimicrobial activity and food applications (Pathania et al., 2018; Liao et al., 2021; Moazeni et al., 2021). Despite their highly potent, lipophilic, and aromatic active ingredients, EOs are not broadly used in the food industry and animal nutrition due to some application limiting factors such as low water solubility, volatility, bioavailability, stability, and limited supply routes.

The most recent nanotechnology advancements can overcome these existing barriers, allowing EOs to be used as low-dose preservatives in food supply chains (Liao et al., 2021). Encapsulation of EOs in nano-emulsions has been revealed as a novel method to increase their stability, efficacy, and commercial viability (El-Sayed and El-Sayed, 2021; Moazeni et al., 2021). Essential oils need to be nano formulated with a small droplet size (20-500 nm) to avoid evaporation and maintain biological function by enhancing cellular uptake and bioavailability of numerous hydrophobic drugs. This results in increasing their solubility, physical stability, and dissolution rate, and bioavailability (Salman et al., 2021; Singh et al., 2021; Ameta et al., 2023).

Exogenous emulsifier supplementation to a broiler diet enriched with high hydrophilic-to-lipophilic balance enhanced body weight and apparent metabolizable energy content while having little influence on fat digestibility (San Tan et al., 2016). Furthermore, when fat was added to broiler diets, nutrient digestibility increased. This could be due to improved emulsion of unsaturated fatty acids and excellent absorption in the digestive tract (Kamran et al., 2020).

Supplementation with dietary encapsulated EOs and organic acids improved growth performance and gut health in Clostridium perfringens-infected broilers by positively modulating the gut microbiota community, strengthening the intestinal barrier function, and differentially regulating enteric immune response (Pham et al., 2020).

The microcapsules had antibacterial and antioxidant properties, and their use as a dietary supplement in broilers challenged with Salmonella Enteritidis at levels of 0.5, 1, and 2 kg/ton improved growth performance, antioxidant status, intestinal microbial residents, ileal morpho-structure, and regulated inflammation and antioxidant genes in ileal tissue (Moharreri et al., 2022). In addition, the gastrointestinal tract microbiome is linked to chicken health and has a substantial impact on the digestion, absorption, and metabolism of feed nutrients in chickens (Lee et al., 2017; Wang et al., 2021). Essential oils as growth-promoting additives reduced feed conversion ratio (FCR), decreased cecal microbes, enhanced nutrient digestibility, and tendency to increase serum protein metabolites of broiler chickens (Irawan et al., 2020).

Magic oil is a novel plant oil nano-emulsion. It is a monounsaturated fatty acids/polyunsaturated fatty acids blend, comprising primarily of linolenic acid (n-3) and linoleic acid (n-6)-oil group (Dhull et al., 2020). The use of nano-emulsified vegetable oil (NEVO) in broiler production diets as a nano-feed additive is intended to boost productivity and produce omega -6 and omega -9 enriched meat. NEVO contains approximately 7% linolenic acid and approximately 50% linoleic acid (Abd El-Hack et al., 2015). Abudabos et al. (2021) conducted a recent study and recommended that NEVO and betaine supplements improve performance, especially at high ambient temperatures in broiler houses. In addition, Suliman et al. (2023a,b) and Al-Garadi et al. (2023b) conducted a recent study and recommended that Magic oil, betaine and/or probiotics be added to the water of growing broiler programs due to favorable meat quality and the most favorable cooking loss value especially under heat stress condition. There are little or no scientific research on supplementing Magic oil with drinking water. Therefore, the priority and novelty of this study to determine the ideal times programs to add Magic oil to drinking water in order to understand how to provide broiler chickens with the best performance results at different stages of supplementation during different growth periods. The researchers aimed to understand how Magic oil, a novel nano-emulsion source of omega-3 and omega-6 polyunsaturated fatty acid as well as probiotics, affected broiler chicken growth performance, ileal histomorphology, carcass characteristics, and serum biochemistry.

MATERIALS AND METHODS

The Ethics Committee of Scientific Research, King Saud University (KSU), Saudi Arabia, accepted this study (Approval No: KSU-SE-21-02).

Experimental Design, Bird's Management, and Tested Products Composition

On arrival, 432-day-old birds (Ross 308) were sexed, weighed individually and divided into 6 treatments with similar body weights. Each group was further distributed to 9 replicates, each with 8 birds (4♂ and 4♀). The experiment was performed in poultry unit under controlled conditions at 22°C to 24°C at the Animal Production Department, KSU (24°43′28.8"N 46°37′07.9"E). The birds were reared in flour cage pens (100 cm width, 100 cm length) under similar managerial and sanitary conditions. The treatment diets were fixed in all experimental groups and formulated to be isocaloric and isonitrogenous to match/exceed the recommendations in the commercial sector in Saudi Arabia. A standard starter (d 0–15) and finisher (d 16–35) diets were offered in mash form based on corn-soy bean meal (Suliman et al., 2023a).

All additives were supplemented to water and were not part of the nutrient matrix. The birds were randomly assigned to 1 of 6 treatment groups upon arrival (Figure 1): T1: Magic oil was supplemented from d 1 to slaughter (d 35). T2: Magic oil was supplemented from d 1 to 4, then from d 17 to 21, and from d 25 to slaughter. In T3 group: Magic oil was supplemented from d 1 to 4, then from d 17 to slaughter. In the T4 group: Magic oil was supplemented from d 1 to 4 and from d 21 to slaughter. Positive control (PC): Probiotic was supplemented from d 1 to 4 and from d 16 to 18. Negative control (NC): no additive (neither Magic oil nor probiotic were supplemented). The periods of adding Magic oil were 35, 20, 23, and 19 d for T1, T2, T3, and T4, respectively. In T2, T3, and T4, the periods of application were specified coinciding with the exposure of birds to stress conditions as post-transportation, vaccination, and adaptation (d 1–4), poster doses vaccination (d 17–21), and finishing period and preparation for slaughter (d 25–35). Usually, nano-emulsified plant oils are used throughout the growing period of broiler chickens, hence the intermittent introduction of Magic oil in this study was also proposed to reduce the total cost of production by reducing the cost of supplementation by nearly half.

Figure 1.

Figure 1

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Experiment design and composition of tested products (Magic oil and Albovit) T1: Magic oil supplementation from 1 to 35 d of age. T2: Magic oil supplementation from 1 to 4, 17 to 21, and 25 to 35 d of age. T3: Magic oil supplementation from 1 to 4 and 17 to 35 d of age. T4: Magic oil supplementation from 1 to 4 and 21 to 35 d of age. Positive control (PC): Probiotic supplementation from 1 to 4 and 16 to 18 d of age. Negative control (NC): Unsupplemented control.

Each liter of Magic oil (Atco pharma, Cairo, Egypt) contains 98.5% nano-emulsified crude oil including 26% monounsaturated fat, 59% polyunsaturated fat (50% linoleic acid; omega-6 & 7% linolenic acid; omega-3;), 14% saturated fats only and vitamin E. Each kg of Albovit (Albafarma, Istanbul, Turkey) contains Enterococcus faecium (3.3 × 1012 CFU), Galacto-oligosaccharides (136,000 mg), Vitamin D3 (200,000 IU), and Vitamin C (200,000 mg).

Performance Measurements

The amount of feed rejected from the amount of feed offered was used to calculate the daily feed intake. Body weight was measured on 5 d basis. Every 5 d, the total feed intake, body weight gain (BWG), and FCR for each group were computed.

Ileal Histology

On d 35, a 1 cm section of the lower ileum from 2 birds per replicate was cut, washed in physiological saline. After that, ileum segments were fixed in 10% formalin for 3 d and then mounted onto slides after cutting up into 5 µm thick slices. The slides were stained with Alcian blue and Hematoxylin & Eosin (H&E stain). Height and width of the villi and histopathological changes were measured as described by Azzam et al. (2020) based on at least 10 well-oriented villi per section using a microscope (IX71 Inverted Olympus) and PC-based image analysis system (Olympus DP72 Microscope Digital Camera; Olympus NV, Aartselaar, Belgium) with software Analysis (Cellsens Digital Imaging Software for Research Application).

Carcass Measurements

On d 35, 18 birds per group (9 ♂and 9 ♀) were randomly selected. After slaughter, the abdominal fat, liver, heart, and gizzard were dissected then and weighed. The yield percentages of the carcass, abdominal fat, liver, heart, and gizzard were calculated relative to live weight (Abudabos et al., 2013). After completion of the experiment, the biological wastes including remaining carcasses and parts were discharged by the guidelines developed by the Labs and Risk Management Unit (ALARM). They were double-wrapped and frozen until disposal. Thereafter, The Safety and Security Committee collected all biological wastes in a specialized room for disposal of chemical and biological wastes.

Blood Biochemistry

By the end of the experiment (d 35), blood samples were collected from exsanguinated birds during Halal slaughter (two 3 mL-sample from 2 birds per each replicate). The collected samples were centrifuged at 3,000 rpm for 10 min. Serum was collected and then transferred to clean plastic tubes, and stored at -20°C until further analysis. Total protein (TP), albumin (ALB), glucose (GLU), triglycerides (TRI), total cholesterol (CHO), alanine transaminase (ALT), uric acid (UA), and aspartate aminotransferase (AST) were analyzed spectrophotometrically (Randox, London, United Kingdom) with reagent kits (Randox, London, United Kingdom) based on the manufacturer's guidelines. Serum globulin = TP - ALB. All the analyses were carried out with duplicate methods.

Statistical Analysis

The data were analyzed by 1-way ANOVA analysis using Statistical Analysis System package version 9.4 software (SAS Institute Inc., Cary, NC) in a general linear model procedure (SAS, 2004). The model used was as follows:

Yij=μ+Ti+eij

Where:

Yij is the observed j variable in the ith treatment;

μ is the overall mean;

Ti is the treatment effects ith; eij is a random residual error.

The PDIFF option was used to test the means for parameters that had significant differences in the analysis of variance. In general, P < 0.05 was chosen as the threshold for statistical significance. All values were expressed as statistical means ± standard error of the mean (SEM).

RESULTS

Growth Performance

The effects of Magic oil supplementation on broiler performance at different experimental periods are shown in Table 1. From d 1 to 4, no differences (P > 0.05) in FI and FCR were detected between the treatments. Body weight gain was significantly (P < 0.05) higher in the T4 Magic oil group than in the NC group and comparable to the PC group. T4 had the highest BWG compared to other Magic oil treatments, T2 and T3 had intermediate BWG, and T1 had the lowest value BWG during the first 3 d of broiler age. It was found that adding Magic oil (particularly T4) and probiotic supplements to birds aged 1 to 4 d during the prestarter period was effective compared to NC.

Table 1.

Effects of water supplementation of nano-emulsified plant-oil and probiotic on broiler performance (Feed intake [g], body weight gain [g], and Feed conversion ratio [FCR]) during 35 d of age (n = 9).

Parameter Treatments1
 SEM P-value
T1 T2 T3 T4 PC NC
1–4
 Feed intake 14.7 14.1 14.6 15.3 15.9 14.5 0.43 0.07
 Body weight gain 13.9b 14.4ab 14.3ab 15.3a 15.4a 13.8b 0.29 <0.001
 FCR 1.06 0.98 1.03 1.00 1.03 1.06 0.03 0.38
4–14
 Feed intake 48.6 49.6 48.4 48.8 49.1 48.3 0.68 0.78
 Body weight gain 37.1 37.1 36.6 37.1 36.0 35.8 0.64 0.49
 FCR 1.31 1.34 1.32 1.32 1.36 1.35 0.02 0.55
14–21
 Feed intake 106.3a 104.1ab 104.4ab 105.5ab 102.1ab 97.8b 1.93 0.04
 Body weight gain 75.3 75.2 76.9 76.7 75.3 71.4 1.46 0.12
 FCR 1.41 1.39 1.36 1.38 1.36 1.37 0.01 0.11
21–30
 Feed intake 148 146 147 148 145 143 1.74 0.31
 Body weight gain 102 99 98 103 101 100 1.68 0.23
 FCR 1.45 1.49 1.52 1.44 1.44 1.44 0.03 0.34
30–35
 Feed intake 176.8 173.0 176.7 179.8 176.9 173.5 4.32 0.89
 Body weight gain 113.2 109.3 112.3 114.7 112.7 112.1 4.41 0.98
 FCR 1.47 1.60 1.58 1.57 1.58 1.55 0.03 0.92
1–35
 Feed intake 3,502 3,463 3,479 3,522 3,459 3,380 43.61 0.29
 Body weight gain 2,429 2,364 2,389 2,480 2,407 2,335 40.58 0.18
 FCR 1.44 1.47 1.46 1.42 1.44 1.45 0.01 0.15
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T1: Magic oil supplementation from 1 to 35 d of age. T2: Magic oil supplementation from 1 to 4, 17 to 21, and 25 to 35 d of age. T3: Magic oil supplementation from1 to 4 and 17 to 35 d of age. T4: Magic oil supplementation from 1-4 and 21 to 35 d of age. Positive control (PC): Probiotic supplementation from 1 to 4 and 16 to 18 d of age. Negative control (NC): Unsupplemented control.

a.b

Means within a row with no common superscript differ significantly (P ≤ 0.05).

When compared to PC and NC, broiler performance did not differ significantly (P > 0.05) between water-supplemented groups at other growing periods. However, when compared to the NC, T1 consumed the most feed (P < 0.05), whereas the other groups consumed similarly from 14 to 21 d of age. From 14 to 21 d of age, all Magic oil groups were fed in the same manner as PC. Overall, T4 followed by T1 of Mgic oil outperformed NC and PC treatments numerically (P > 0.05) during the entire experimental period (1–35 d of age).

Ileal Histomorphology

As shown in Table 2, the ileal morphometry data of the different treated groups presented that T1 had the longest ileal length, T2 had the intermediate length, and T3 and T4 had the shortest (P < 0.001), compared to the PC and NC groups.

Table 2.

Effects of water supplementation of nano-emulsified plant-oil and probiotic on intestinal histology of broiler chickens at 35 d of age (n = 9).

Water supplementation treatments1
 SEM P-value
T1 T2 T3 T4 PC NC
Villi length (μm) 506.4a 453.6b 396.7c 393.9c 427.0bc 404.9c 10.4 <0.001
Villi width (μm) 76.07a 70.22ab 71.66a 58.10c 78.98a 60.96bc 2.34 <0.001
Villi surface area (mm2) 0.12a 0.10b 0.09bc 0.07d 0.10b 0.08cd 0.004 <0.001
Crypt-depth (μm) 33.70c 34.04c 38.37bc 45.35a 43.45ab 27.78d 1.45 <0.001
Villi length/ crypt depth 15.67a 13.89a 10.63b 9.54b 10.62b 14.73a 0.52 <0.001
Goblet cells numbers 150.73a 122.03bc 102.70cd 128.78b 118.73bc 93.30d 4.76 <0.001
GCD/100 µm villus area 14.89ab 13.51bc 13.06bc 16.43a 14.24b 11.64c 0.53 <0.001
Goblet cells scores3 3.0ab 2.7bc 2.6bc 3.3a 2.8b 2.3c 0.11 <0.001
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T1: Magic oil supplementation from 1 to 35 d of age. T2: Magic oil supplementation from 1 to 4, 17 to 21, and 25 to 35 d of age. T3: Magic oil supplementation from 1 to 4 and 17 to 35 d of age. T4: Magic oil supplementation from 1 to 4 and 21 to 35 d of age. Positive control (PC): Probiotic supplementation from 1 to 4 and 16 to 18 d of age. Negative control (NC): Unsupplemented control.

abcd

Means within a row with no common superscript differ significantly (P ≤ 0.05).

3

Goblet cells scores= (goblet cell density per 100 µm villus area)/ 5 µm of cutting thick slices.

Abbreviation: GCD, goblet cell density.

The T1, T3, and PC groups had the greatest ileal width, whereas the other groups did not differ significantly from the NC group. When compared to the NC, the villus surface area of the ilium of birds increased significantly (P < 0.001) in T1, followed by both groups T2 and PC. Compared to the NC, the crypt-depth of ilia of birds increased significantly (P < 0.001) in group T4, followed by group PC, then groups T2, T3, and T4. The villus length/crypt depth ratio increased (P < 0.001) in groups T1 and T2 compared to T3, T4, and PC groups, but did not differ in comparison to NC group. Goblet cells were highest (P < 0.001) in treatment 1 and lowest in NC, with intermediate numbers in other groups. Goblet cell density (GCD)/100 µm villus area revealed a higher density (P < 0.001) in group T4, and lowest in NC, with intermediate density in other groups. On the other hand, goblet cells scores revealed a higher score (P < 0.001) in group T4 (3.3), and lowest in NC (2.3), with intermediate scores in other groups.

The results of the histomorphology examination of the bird's intestine in the different experimental groups are shown in Figure 2. NC with enterocyte detachment and moderate goblet cell proliferations. T1 had goblet cell hyperplasia with mild ileal villus enterocytes and parts in their Lumina. T2 demonstrates free lumen with fewer goblet cells and no defects on their enterocytes. T3 showing an ileal villus sided with goblet cell proliferation. T4 showed thickened villi with congested blood capillaries of the lamina propria with mild goblet cell proliferation. PC group revealed a villus that was arranged higher and wider, with prominent goblet cell hyperplasia.

Figure 2.

Figure 2

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Broiler intestines after water supplementation of nano-emulsified plant-oil and probiotic. Stained with Alcian blue and H&E stain (×200). NC. Negative control, no additive, demonstrating enterocyte detachment with moderate goblet cell proliferation. T1. Magic from d 1 to slaughter, demonstrating goblet cell hyperplasia with mild ileal villus enterocytes and parts in their Lumina. T2. Magic from 1 to 4 d then from 17 to 21 d and from 25 to slaughter, demonstrating free lumen with a decreased number of goblet cells and no defects on their enterocytes. T3. Magic from 1 to 4 d, then 17 d to slaughter, demonstrating an ileal villus sided with goblet cell proliferation. T4. Magic from 1 to 4 d and from 21 to slaughter, showing thickened villi with congested blood capillaries of lamina properia with mild goblet cell proliferation. PC. Positive control: Probiotics from 1 to 4 d and 16 to 18 d, showed that a villus arranged higher and wider, with marked goblet cell hyperplasia.

Carcass Traits

As shown in Table 3, there were no significant differences in carcass traits and carcass yield of broilers at 35 d between birds that received Magic oil water supplementation at different growing periods when compared to PC and NC.

Table 3.

Effects of water supplementation of nano-emulsified plant oil and probiotic on carcass measurements of broiler chickens at 35 d of age (n = 18).

Treatments1
 SEM P-value
T1 T2 T3 T4 PC NC
Live weight (g) 1,944 1,981 1,933 1,918 1,950 1,964 29.4 0.71
Hot carcass weight (g) 1,402 1,433 1,389 1,388 1,417 1,426 23.3 0.65
Carcass yield (%) 72.1 72.3 71.9 72.3 72.7 72.6 0.40 0.63
Abdominal fat (%) 1.40 1.40 1.40 1.30 1.50 1.50 0.10 0.61
Liver (%) 2.40 2.30 2.20 2.20 2.30 2.3 0.10 0.46
Heart (%) 0.60 0.60 0.60 0.60 0.60 0.6 0.001 0.39
Gizzard (%) 1.80 1.60 1.70 1.80 1.60 1.7 0.10 0.30
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T1: Magic oil supplementation from 1 to 35 d of age. T2: Magic oil supplementation from 1 to 4, 17 to 21, and 25 to 35 d of age. T3: Magic oil supplementation from1 to 4 and 17 to 35 d of age. T4: Magic oil supplementation from 1 to 4 and 21 to 35 d of age. Positive control (PC): Probiotic supplementation from 1 to 4 and 16 to 18 d of age. Negative control (NC): Unsupplemented control.

The relative weights of abdominal fat, liver, heart, and gizzard were calculated relative to live weight.

Blood Biochemistry

Table 4 summarizes the effects of Magic oil and probiotic supplementation on blood biochemical indices (TP, ALB, globulin, UA, GLU, CHO, TRI, ALT, and AST) at 35 d of age. According to the statistical analysis, the treatment groups had no significant effects on the blood biochemical parameters of broiler chickens. This meant that the blood parameters were all within normal ranges.

Table 4.

Effects of water supplementation of nano-emulsified plant oil and probiotic on blood biochemical indices of broiler chickens at 35 d of age.

Treatments1
 SEM P-value
T1 T2 T3 T4 PC NC
Total protein 2.76 2.94 2.94 2.97 2.94 2.80 0.10 0.52
Albumen 1.34 1.35 1.31 1.36 1.37 1.30 0.05 0.85
Globulin 1.42 1.60 1.63 1.61 1.56 1.50 0.08 0.43
Uric acid 3.14 2.72 2.64 2.54 2.70 2.49 0.25 0.53
Glucose 236.19 233.9 227.7 242.48 229.83 232 8.35 0.87
Cholesterol 174.95 172.92 168.01 175.53 155.61 158 8.97 0.47
Triglycerides 106.08 103.03 96.17 99.78 91.87 106.2 6.37 0.55
ALT 9.23 10.33 8.97 9.4 8.53 8.88 0.52 0.22
AST 302.83 263.73 282.04 283.33 276.14 282.06 13.11 0.47
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T1: Magic oil supplementation from 1-35 d of age. T2: Magic oil supplementation from 1 to 4, 17 to 21, and 25 to 35 d of age. T3: Magic oil supplementation from1 to 4 and 17 to 35 d of age. T4: Magic oil supplementation from 1 to 4 and 21 to 35 d of age. Positive control (PC): Probiotic supplementation from 1 to 4 and 16 to 18 d of age. Negative control (NC): Unsupplemented control. n = 12 blood samples.

Abbreviations: ALT, alanine transaminase, AST, aspartate transaminase.

DISCUSSION

Poultry production is currently in high demand around the world. Performance parameters are optimized for this purpose, such as fast-growing broiler chickens with low feed consumption and a better flock health status. Because of the increased demand, many antibiotic-free products are now being used. As there is increased pressure to reduce the amount of antibiotics used as bacteriostatic or bactericidal agents for poultry, there is an urgent need for novel solutions to maintain poultry productivity and efficacy. Among the substitutes, EOs have enormous potential because they are natural, free of hazardous deposits and chemicals, and less toxic. Essential oils are plant-based extracts, and there are approximately 3,000 known EOs, 300 of which have been identified as useful and commercially important. It has been demonstrated that EOs have extensive in vitro and in vivo research to produce special effects on a wide range of pathogens (Raza et al., 2022).

Dietary palm oil at 4% improved feed intake, and dietary inclusion of palm oil at 2 and 3% increased live weight by 4% and 3%, respectively, over the control (Rahman et al., 2010). They concluded that profitable broiler production without palm oil supplementation might be possible due to similar earned profit from each group. Increased dietary fat levels such as palm oil at 5% had no effect on daily BWG and feed consumption (Rahman et al., 2010). Less performance of broiler chickens (Rahman et al., 2010) or rabbits (Fernández-Carmona et al., 1998) perhaps attributed to the negative digestive influences of vegetable oil-based feeds with other nutrients such as starch, resulting in lower intake and output. Adding NEVO to broiler chicken drinking water at a rate of 5 mL/L increased average daily gain by 5.7%, performance efficiency factor by 7.7%, and FCR by 6.1% over control during d 21 to 35, while feed consumption remained unchanged (Abudabos et al., 2021). In rabbits, NEVO at 1.0 mL/L of water increased growth more than NEVO at 0.5 mL/L of water (Ghanima et al., 2021). According to some studies, broiler chickens can use considerable amounts of nutritive fat as an energy source (Rahman et al., 2010; Singh and Gaikwad, 2020).

In this study, the addition of Magic oil especially from d 1 to 4, revealed improvement in birds BWG. As a result, a good start leads to a good finish. Along the experiment period (1–35), broiler chickens in the T4 group of the Magic oil supplemenation program consumed feed at a rate 4.20% and 1.82% higher than those in the NC and PC groups, respectively, followed by those in the T1 (3.61%), and the T2 and T3 were intermediate (2.46% and 2.93% higher than those in the NC, respectively). Furthermore, broiler chickens in the T4 of Magic oil outperformed BWG in those in the NC by 6.21% and in the PC by 3.08%, followed by those in the T1 (4.03%), whereas those in the T2 and T3 were intermediate (1.24% and 2.31% higher than NC, respectively). Consequently, T4 of Magic oil improved feed conversion at a rate that was 2.07% compared to NC and 1.39% compared to PC, followed by T1 (0.69%). These findings are consistent with the fact that dietary inclusion of nanoencapsulated EOs improved bird body weight, as demonstrated by Hosseini and Meimandipour (2018), who investigated the effect of thyme oil encapsulated with chitosan on broiler performance, increasing BWG and FCR at different growing periods. In addition, the dietary inclusion of broiler ration with EO and probiotics resulted in an enhancement of BWG of broiler chickens (Abd El-Hack et al., 2021; Abd El-Hack et al., 2022). It was reported that broilers supplemented with NEVO, consisting of polysorbate-80, vitamin E, and soybean oil, effectively improved growth performance parameters under heat stress conditions (Abudabos et al., 2021). In ovo administration of nano encapsulated thyme oil was increase in performance of broiler chicken (Yaseen et al., 2022). In line with Nobakht et al. (2011) who observed that the addition of sunflower, canola, and soybean oils to the diets of chicks had no effect on their liver weights. On d 1 to 35, the nano emulsion of the EOs could increase BWG and FCR in comparison to conventional EOs, implying that Magic oil can efficiently increase BWG and FCR of the broilers due to stimulation of the digestive and immune systems by the active compounds in plant oil. The effect of the NEVO supplement have improved the final BWG, and FCR of rabbit (Ghanima et al., 2021).

The morphometric histologic characterization of the bird intestine on d 35, on the other side, revealed that the villi length, villi width, villi surface area, and goblet cell numbers were significantly higher in the T1 of Magic oil than in the NC by 25.07, 24.79, 50, and 61.6%, respectively. Crypt-depth, GCD/100 µm villus area, and goblet cell scores were improved in the T4 of Magic oil by 63.3, 41.2, and 43.48%, respectively, compared to the NC. The villi of small intestinal are lined with a variety of epithelial cells, including goblet cells and enterocytes that are derived from crypt stem cells and are vital for nutrient uptake and host defense (Carulli et al., 2014; Reynolds et al., 2020). Goblet cells in chickens, secrete acidic mucin 2, which is a major component of the mucus that lines the intestinal tract and serves as a protective barrier between epithelial cells of the intestine and pathogens, and is thus essential for chick health (Reynolds et al., 2020). When goblet cells produce the protective mucus layer during the establishment of the intestinal microbiome, they reduce bacterial density (Apajalahti et al., 2004).

Probiotics-treated groups revealed significant improvement in intestinal morphometric parameters including intestinal length, width, and villus surface area followed by Magic oil supplied groups (T1 and T4). While, the crypt-depth, villus length/crypt depth, goblet cells number, and GCD/100 µm villus area revealed a significant increase in Magic oil-treated groups followed by PC group in comparison with control untreated birds in the NC group. These outcomes were parallel to Bogusławska-Tryk et al. (2021) who confirmed that in-ovo administration of prebiotics, probiotics, and synbiotics revealed a positive impact on the production parameters and intestinal morphology in broiler chickens. Also, Jadhav et al. (2015) concluded that the improvement in the intestinal histologic morphometric parameters enhances nutrient absorption which is later reflected in the improvement of BWG of treated birds.

Incorporating probiotics, prebiotics, and/or phytogenics into chicken diets is an efficient way to support growth and optimize digestive tract health in poultry (Shehata et al., 2022). Furthermore, the early development of duodenal and ileal morphologic and functional traits of broilers in the starter stage improves early growth performance (Jha and Kim, 2021). Probiotics and Magic oil may alter the microbiota of the gut and improve the integrity of epithelial cells of the intestine, resulting in improved nutrient absorption and thus growth performance.

The program of adding Magic oil from d 1 to 35 of age leads to an increase in the number of goblet cells. In the current study, a noticeable increase in crypt depth was noticed in the program of adding Magic oil from 1 to 4 and 21 to 35 d of age. The increase in crypt depth may be satisfactory due to stem cells proliferating in the crypt, potentially increasing the number of mucin protein-producing goblet cells and enterocytes (Hutsko et al., 2016).

Supplemented water with Magic oil from 1 to 4 and 21 to 35 d decreased ileum villi length, villus width, villus surface area, and villus length: crypt to depth ratio. This finding may be due to irritation of intestinal tissues (Windisch et al., 2008; Zeng et al., 2015). The intestinal villus is related to a bird's ability to absorb nutrients from its diet (Wang et al., 2015). The absorptive area may increase by increasing villus length, resulting in improved digestive enzyme activity and increase transport of nutrients at the villus surface (Tufarelli et al., 2010). The intestinal villus can familiarize and react morphologically to feed micro and macrostructure factors (Laudadio et al., 2012; Ege et al., 2019). In agreement with Ege et al. (2019), a greater villus width, villus length, and villus length/crypt depth ratio indicated that larger digestive and absorption areas were noted in the birds fed Magic oil from 1 to 35 d of age compared to control.

Nano-emulsified plant oil and probiotic have significantly improved gain body weight at 1 to 4 d of age and numerically promote overall broiler growth compared to the control. This may be attributed to enhancing goblet cells and absorptive surface area may result in increasing nutrient absorption. Nonetheless, improving nanoemulsion use in broiler diet by improving target-specific nanoparticles to improve broiler growth, physiological parameters, and intestinal absorptive function is required. There is an imperative need to employ nano-emulsified plant oil, which has the potential to improve bioavailability mechanisms. The function of nano-emulsified beneath plant EO in terms of transportation, toxicity, and bioavailability to the intestinal ecosystem is still being researched. As well as nano-emulsified plant oil needed further study with various doses to detect suitable levels that have the potential promote broiler growth and development. This could be attributed to the presence of bioactive substances that promote nutrient digestion and absorption. Similar results confirmed the association between botanicals or their derivatives in bird diets and good digestive health via antioxidant and antimicrobial activities, which could improve digestion, improve liver function, and promote pancreatic enzyme secretion in the intestine (Langhout, 2000; Seidavi et al., 2022). Improved efficiency of nutrient metabolism could lead to improved growth performance indices. However, performance criteria cannot always be used to determine the impacts of phytogenic supplements or their active ingredients (Lee et al., 2003). In addition, plant-derived active components such as tannins, flavonoids, and saponins have been used in many in vitro and in vivo studies (Kostadinović and Lević, 2018; Abdelli et al., 2021; Iyayi et al., 2021; Agubosi et al., 2022). They found that coconut, sunflower, bitter, neem, or their extracts may have antimicrobial, antioxidant, anti- and inflammatory activities.

Data suggest that an increase in absorption area may be related to an increase in the number of goblet cells and consequently increases feed intake in the group receiving Magic oil, especially at 1 to 35 d of age, but does not necessarily improve feed conversion efficiency. Further research is required to find out an optimal marketing age in terms of economic considerations to calculate the costs and profits at different marketing ages (Abougabal, 2020). Broiler chickens supplemented with Magic oil from 1 to 35 d had increased feed intake at 4 to 21 d as well as increased absorption villus surface area and goblet cells. Furthermore, birds supplemented with watery Magic oil, particularly from 1 to 4 and 21 to 35 d of age, as well as probiotics, gain more body weight during the first 4 d of age.

It was found that dietary nano-emulsified plant oil supplements had no impact on dressing, abdominal fat, or gizzard, which was in agreement with Abudabos et al. (2021). Moreover, these results are consistent with those of Nisar et al. (2021), who revealed that dietary supplementation did not affect carcass traits. T4 attempted (P > 0.05) to reduce abdominal fat by 13.33% compared to controls. High dietary energy EOs are improved carcass traits (Ghanima et al., 2021; Pirastehfard et al., 2021). Dietary nano-thyme oil was not affected the carcass traits of broiler chicken (Yaseen et al., 2022).

The obtained results here revealed that the water supplemented with Magic oil has no impact on blood biochemical indices compared with PC and NC at 35 d of age. These findings confirmed that both Magic oil and probiotics are safe to be administrated by the birds, as they revealed no change in blood chemistry at the age of marketing. These results agreed with Midilli et al. (2004) who observed that feeding of broilers to probiotics had no impact on serum TP in the treated groups. In addition, Mathivanan and Kalaiarasi (2007) noticed that the serum alkaline phosphatase, ALT, AST, and TP levels in broilers treated with probiotics did not differ from the control. The addition of nano-boron to the ration can lead to improvement in some blood parameters of the broiler (Ali and Abbas, 2022). Biochemical blood tests are used to assess the physiological, nutritional, and pathologic status of animals. Our outcomes agree with those of Ghanima et al. (2021) who observed that there was no change in plasma levels of ALB or TRI in rabbits fed 1 mL NEVO/L; however, they disagree with those of Ghanima et al. (2021), who found that 1 ml NEVO /liter decreased total lipids and total CHO while increasing TP and globulin. Dietary supplementation with sunflower oil as a vegetable source did not affect plasma total lipids and CHO levels in rabbits, according to Fekete et al. (1990). ALT and AST levels were used to determine the normal liver function (Ghasemi-Sadabadi et al., 2021), and no changes in this study indicated that NEVO or probiotics were safe for liver function. The medicinal properties of many plants are linked to secondary metabolites such as EOs (Abd El-Hack et al., 2015). The addition of a nano Moringa leaves extract to drinking water resulted in a noteworthy increase in TP, globulin, and ALB concentrations. While there were no significant differences in UA and GLU concentrations across all experimental treatments (Oraibi and Ali, 2021). A significant improvement in serum metabolites was noted with increasing NEVO levels. A remarkable increase in the serum TP and globulin associated with a substantial decrease in the serum total lipids and total CHO levels was also noticed with increasing NEVO levels (Ghanima et al., 2021).

Here, the water supplements, the nano-emulsified, and the Albovit probiotic were used as PC and NC groups, respectively. Overall, birds in the T4 group of the Magic oil supplementation program along the experiment (1–35) consumed 4.20% more feed than the NC, gained 6.21% more than the NC, and improved feed conversion by 2.07% more than the NC. On the other hand, NEVO could be compared with antibiotics, probiotics, or conventional EOs to test the possibility of using it as a replacer for these materials. In the T4 group of the Magic oil supplementation program, the birds consumed 1.82% more food and gained 3.08% more than the Albovit group. Moreover, the conversion of feed to meat was improved by 1.39% more than Albovit group. It is recommended that further studies have to compare plant products and/or EOs with antibiotics, probiotics, prebiotics, and symbiotic agents of unknown mechanisms to estimate their actual values to growing broilers. It is also advisable to approach nano-emulsified plant oils against conventional ones to estimate their actual usefulness in poultry production.

CONCLUSIONS

In conclusion, the water supplementation of Magic oil particularly throughout the growing period (T1: d 1–35) and T4 (from d 1 to 4 then from d 21 to 35) improved BWG from 1 to 4 d as probiotic and feed intake from 14 to 21 d, enhanced the intestinal morphometric, improved goblet cell proliferation similar (T1, T2, and T3) or better (T4) than probiotic. No significant effect was observed for the water supplementation of Magic oil and probiotics on carcass measurements and blood biochemical indices of broiler chickens at 35 d of age. Further studies are needed to evaluate the effect of adding both nano-emulsified plant oil and probiotics on different parameters.

Acknowledgments

ACKNOWLEDGMENTS

This research was funded by the Deputyship for Research & Innovation, Ministry of Education in Saudi Arabia for funding this research work through the project no. (IFKSURG-2-432).

The authors extend their appreciation to the Deputyship for Research & Innovation, Ministry of Education in Saudi Arabia for funding this research work through the project no. (IFKSURG-2-432).

Institutional Review Board Statement: The study was approved by the Ethics Committee of Scientific Research, King Saud University (KSU), Saudi Arabia (Approval No: KSU-SE-21-02).

Data Availability Statement: The data presented in this study are available on request from the corresponding author.

Authors' Contributions: Conceptualization, G.S. and E.H.; methodology, A.A., G.S., and E.H.; software, H.A.; validation, M.A., G.S,. and E.H.; formal analysis, M.A.; investigation, A.A., G.,S. and E.H.; resources, A. A.; data curation, H.B-A.; writing—original draft preparation, M.Q., G.,S. and E.H.; writing—review and editing, A.S.; visualization, G.S.; supervision, A.S.; project administration, G.S., and A.A.; funding acquisition, A. A. All authors have read and agreed to the published version of the manuscript

DISCLOSURES

The authors declare no conflicts of interest.

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