{"paper_id":"2a2f0735-b0fd-44f6-bbc6-7b0bf7fe0192","body_text":"Impact of in ovo feeding of nano-encapsulated garlic essential oil on broiler hatch percentage, performance, intestinal development, and blood parameters | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Impact of in ovo feeding of nano-encapsulated garlic essential oil on broiler hatch percentage, performance, intestinal development, and blood parameters Negin Amiri, Mohsen Afsharmanesh, Mohammad Khajeh Bami This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7119473/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 9 You are reading this latest preprint version Abstract The purpose of this research was to determine the impact of in-ovo injection of garlic essential oil (GEO) on post-hatch broiler chicken performance, intestinal development, hatch percentage, and blood parameters. 360 Ross 308 eggs weighing 62 ± 2 g were selected and 15 eggs per replicate were used in a completely randomized design. The experimental treatments were as follows: (1) control or non-injected, (2) in-ovo injection of physiological saline or positive control, (3) in-ovo injection of 0.1 free GEO, (4) 0.2 free GEO, (5) 0.1 nanoencapsulated GEO, (6) 0.2 nanoencapsulated GEO into the yolk sac at day 7 of incubation period. In contrast to the control treatment (injection-free), the results demonstrated that in-ovo injection of free GEO lowered hatch percentage (P < 0.05). The post-hatch broiler chicken performance was unaffected by different treatments. The breast glycogen concentration and the villus height and villus height to crypt depth ratios were higher in the 0.1 ml of nanoencapsulated GEO group (P < 0.05). The in-ovo injection of nanoencapsulated GEO decreased E. coli counts compared to control group (P < 0.05). In conclusion, chitosan nanoencapsulation has the potential to protect bioactive substances and enhance the benefits of GEO in newly hatched chickens. In-ovo injection Yolk sac Garlic essential oil Nano-encapsulation Broiler chicken Introduction The poultry meat and egg are widely regarded as suitable sources of nutrients, particularly proteins and during the past several decades, a lot of people have become interested in using poultry raised organically. Additionally, the search for new antimicrobial agents has been prompted by the rise in antibiotic resistance as well as unfavourable side effects of some antibiotic medications (Yazdanpanah Goharrizi and Tasharofi 2017 ). Various plant parts and their essential oils (EO) are recognized for their antibacterial properties and advantageous biological effects (Dehghani et al. 2018 ). One of the most important and beneficial plants used for medicine is garlic ( Allium sativum ). The plant has been used for many centuries to treat intestinal parasites as well as a variety of fungal, viral, and bacterial infections in various nations to prevent health problems (Abd El-latif et al. 2013 ). Diallyl disulfide, allyl methyl trisulfide, allyl (E)-1-propenyl disulfide, and allyl methyl disulfide were all included in the chemical makeup of GEO (Satyal et al. 2017 ). Contrarily, nanotechnology is a novel technology that can be used to improve food quality and the effectiveness of food production, storage, processing, transportation, traceability, and safety (Mahmoud 2012 ). It is expected that the possibilities will change as a result of nanotechnology. Increased surface area to volume due to nanoprocessing improves absorption and reduces the need for supplements (Jafari et al. 2022 ). Nanoencapsulation, a novel technique to create the potential for a better impression of the covered materials, is the application of nano coating to various materials. Combining these two hypotheses shows that using plant EO and nanotechnology yields better results. Therefore, the purpose of this research was to determine the impact of in-ovo injection of GEO into the yolk sac on post-hatch broiler chicken performance, intestinal development, hatch percentage, and blood parameters. Materials and methods Preparation of materials Volatile oil was distilled from the ground plant material using Clevenger distillation apparatus (Herbal Exir Co., Mashhad, Iran). Ethanol and acetic acid were provided by Merck (NYSE: MRK). Nanoencapsulation were produced using chitosan, derived from crab shell and sodium triphosphate pentabasic (TPP) and provided from Sigma Chemical Company (St. Louis, MO.). Nanoencapsulation of the GEO was performed by ionic gelation according to the procedure of Sawtarie et al. ( 2017 ). Chitosan was dissolved at a concentration of 1 mgmL − 1 in 1% (w = v) acetic acid and sonicated before the solution became transparent. The dropwise addition of 10mL TPP solution (1 mgmL − 1 ) to a 25mL chitosan solution (pH = 5), under constant stirring at room temperature, produced chitosan-TPP nanoparticles by ionic gelation. For the preparation of chitosan-TPP nanoparticles loaded with herbal EO, 20% (w = v) EO were added to the chitosan solution before adding the TPP solution. Design and animals A total of 360 fertile eggs (flock with 38 week) with mean weight 62 ± 2 g (Ross 308) were used in this 5-week trial (3 weeks of incubation and 2 weeks of growing period). Eggs were randomly assigned to 6 treatments with 4 replications of 15 fertile eggs each. The experimental treatments involved: 1) control or non-injected, 2) in-ovo injection of 0.1 ml physiological saline or positive control, 3 and 4) in-ovo injection of 0.1 and 0.2 ml free GEO, 5 and 6) in-ovo injection of 0.1 and 0.2 ml nanoencapsulated GEO. Setters with automatic temperature control (37.5 ± 0.1) and rotation of the eggs every 1 hr were maintained at 60% relative humidity until day 18 of incubation, when eggs were transferred to hatcher with 70% relative humidity, the egg rotation was stopped. After hatch, newly-hatched-chicks were kept in pens for 2 weeks. Diet was formulated with UFFDA based on corn and soybean meals (Table 1 ) and the chicks had ad libitum access to water and feed (formulated according to the standards prescribed in Ross 308 Broiler Nutrition Specification) (Aviagen 2014 ). Table 1 Ingredient and composition of the basal diet. Item Amount Ingredients (%) Corn 523.70 Soybean meal 390.00 Corn gluten 25.00 Soybean oil 16.50 Calcium carbonate 12.50 Dicalcium phosphate 16.00 Salt 2.50 Sodium bicarbonate 2.00 DL-Methionine 3.20 L- Lysine HCL 2.50 L-Threonine 1.00 Vitamin and mineral premix 1 5.00 Phytase 10000 (hostazym-p) 0.05 Calculated chemical composition Metabolisable energy (MJ/kg) 12.20 Crude protein 220.50 Methionine 6.28 Methionine + cysteine 9.25 Lysine 12.60 Threonine 8.38 Tryptophan 2.28 Arginine 13.60 Calcium 10.10 Available phosphorous 5.03 Sodium 1.70 Linoleic acid 19.40 1 Provided per kg of diet: retinyl acetate, 10000 IU; cholecalciferol, 4500 IU; DL-α-tocopheryl acetate, 65 IU; menadione, 3 mg; thiamin, 2.5 mg; Riboflavin, 5.2 mg; Niacin, 35 mg; Pantothenic Acid, 18 mg; pyridoxine, 3.2 mg; Biotin, 0.18 mg; Folic acid, 1.6 mg; cyanocobalamin, 0.017 mg; choline choloride, 800 mg and antioxidant, 2.5 mg. Provided per kg of diet: copper, 16 mg; iodine, 1.25 mg; iron,20 mg; manganese,120 mg; selenium, 0.30 mg and zinc, 100 mg. In-ovo injection procedure The injection site was disinfected with 96% alcohol, and then, 0.1 and 0.2 ml of each solution was injected into the yolk sac at day 7 of incubation period, using a 23-gauge needle with depth of 28 mm from the broad end of the egg. Then, the holes were sealed using melt paraffin. Data collection The hatch percentage and chicken weight on the first day for every replicate was recorded and then experimental period lasted 14 days. The feed amounts offered and refused were measured at days 7 and 14 for each pen to calculate feed intake (FI). Also, chicks were weighed at above-mentioned days to calculate body weight gain (BWG) changes. By having these two measurements, feed conversion ratio (FCR) was calculated as feed intake consumed per unit of weight gained. At days of hatch and 7, two chicks from every replicate were slaughtered to measure glycogen content of liver and breast and jejunum morphology. Within 1 min after the birds were killed, liver and breast were taken in plastic bags and frozen in -20°C until the start of procedure. Afterwards, 0.2 g samples of liver and breast were added to 1 ml of 8% perchloric acid, homogenized (in ice) for 1 min and centrifuged at 14,000 × g at 4°C for 30 min. The supernatant was removed, and 1 ml petroleum ether (60–80°C) was added. After being mixed, the petroleum ether fraction was removed, and samples from the bottom layer were transferred to a new tube for glycogen determination. For histopathologic analysis, 2-cm tissue samples from the middle of jejunum were obtained and fixed in 10% buffered formalin (100 ml of 40% formaldehyde, 4 g phosphate, 6.5 g dibasic sodium phosphate and 900 ml of distilled water) for 24 hr, and then, the 10% buffered formalin was renewed. Tissues were dehydrated by transferring through a series of alcohols with increasing concentrations, placed into xylol and embedded in paraffin. A microtome was used to make five cuts that were 5 µm. The cuts were stained with haematoxylin–eosin. The values were measured using a light microscope. Measurements of villus height and width and crypt depth were determined at a magnification of 10X. A minimum of five measurements per slide were made for each parameter and averaged into one value (Tako et al. 2004 ). The contents of ileum (10 cm anterior to the junction with caecum and rectum) were collected from 60 birds and put into sterilized vials, and then went for counting the microbial population such as E. coli and Lactobacilli which transported immediately to the laboratory. Population of E. coli and Lactobacilli were estimated as CFU g − 1 using subsequent dilution method. Bacterial counts were performed and cultured using MacConkey agar (at 37°C in an incubator for 1 d) for E. coli and De Man, Rogosa and Sharpe (at 37°C in an incubator for 2 d) for Lactobacilli (Yang et al. 2012 ). Blood samples were obtained from chicks mentioned above and drawn into tubes. After coagulation, the serum samples were centrifuged at 4000 rpm for 10 min, and analysis of serum glucose and cholesterol contents was carried out by enzyme procedure of CHOD-PAP and biochemistry kit (Asan Pharm. Co., Korea) (Richmond 1973 ). Statistical analysis All data were subject to analysis of variance following the GLM procedure in a completely randomized design using the SAS software program. The difference among treatments was compared using the Duncan’s multiple range test. The treatment effect was considered significant at a probability value below 0.05. Results Hatchability and body weight The effects of injecting several types of GEO in ovo on hatchability and chick body weight at hatching are shown in Table 2 . The results showed that in-ovo administration of GEO had a substantial impact on hatchability but experimental treatments had no significant impact on the body weight of the chicks (P > 0.05). The free GEO groups (0.1 and 0.2 ml) had the lowest hatchability, whereas the control treatment (without injection) had the highest. The lack of a noticeable difference between the treatments containing encapsulated GEO and the control group is interesting (without injection). Table 2 Effects of in ovo feeding of garlic essential oil (GEO) on hatchability and hatched chick weight. Treatment Weight (g) Hatch (%) Control (non-injected) 43.31 89.50 a 0.1 ml physiological saline 43.93 82.75 bc 0.1 ml free GEO 43.71 79.50 c 0.2 ml free GEO 43.80 81.00 c 0.1 ml nano-encapsulated GEO 44.06 88.00 ab 0.2 ml nano-encapsulated GEO 43.48 87.50 ab SEM 0.182 1.26 P -value 0.085 0.000 a,b,c Values within a column with different superscripts differ significantly at P < 0.05. Performance Table 3 shows the impact of in-ovo feeding different types of GEO on the performance of broiler chickens. The findings show that experimental treatments had no effect on BWG, FI, or FCR (P > 0.05). Table 3 Effects of in ovo feeding of garlic essential oil (GEO) on growth performance of broilers (1 to 14 d). Treatment Body weight gain (g/b/d) Feed intake (g/b/d) Feed conversion ratio (g/g) 1–7 d 8–14 d 1–7 d 8–14 d 1–7 d 8–14 d Control (non-injected) 147.1 344.4 149.3 442.3 1.01 1.28 0.1 ml physiological saline 147.2 337.6 147.9 440.9 1.00 1.30 0.1 ml free GEO 147.7 339.1 148.2 441.2 1.00 1.30 0.2 ml free GEO 147.9 338.4 150.6 443.6 1.01 1.31 0.1 ml nano-encapsulated GEO 151.0 348.5 148.2 440.5 0.982 1.26 0.2 ml nano-encapsulated GEO 150.2 348.2 148.8 440.8 0.990 1.26 SEM 1.50 3.11 1.35 1.36 0.014 0.012 P -value 0.344 0.064 0.739 0.583 0.496 0.067 Glycogen content of liver and pectoral muscle The concentrations of glycogen in the breasts of 7-day-old chicks were significantly influenced (P < 0.05) (Table 4 ). So, the level of breast glycogen was highest in the 0.1 nanoencapsulated GEO group, whereas the lowest level was observed in the control group. Liver glycogen concentration at hatch and 7 days old were not significantly different between treatments. Table 4 Effects of in ovo feeding of garlic essential oil (GEO) on glycogen levels in both liver tissue and breast muscle of broilers. Treatment Breast glycogen Liver glycogen 0 d 7 d 0 d 7 d Control (non-injected) 0.153 0.280 b 0.136 1.04 0.1 ml physiological saline 0.214 0.302 ab 0.122 0.953 0.1 ml free GEO 0.214 0.292 ab 0.190 0.946 0.2 ml free GEO 0.235 0.359 ab 0.173 0.989 0.1 ml nano-encapsulated GEO 0.237 0.401 a 0.174 1.06 0.2 ml nano-encapsulated GEO 0.235 0.391 ab 0.165 1.14 SEM 0.027 0.025 0.024 0.053 P -value 0.284 0.009 0.383 0.134 a,b Values within a column with different superscripts differ significantly at P < 0.05. Jejunum morphology and microbial population The effect of in-ovo feeding of different forms of GEO on intestinal morphology and the microbial population of broiler chickens is presented in Table 5 . Based on the results, the villus width, crypt depth, and lactobacillus bacterial population were not influenced by experimental groups (P > 0.05). However, villus height, villus height to crypt depth ratios, and E. coli bacterial population were significantly affected (P < 0.05). The villus height and villus height to crypt depth ratios were considerably higher in the 0.1 ml of nanoencapsulated GEO group compared to control group (P < 0.05). Also, the in-ovo injection of nanoencapsulated GEO decreased E. coli counts compared to both control groups (P < 0.05). Table 5 Effects of in ovo feeding of garlic essential oil (GEO) on intestinal morphology and microbial population of broilers. Treatment Villus height µm)) Villus width (µm) Crypt depth µm)) Villus height/crypt depth Lactobacillus E.coli Control (non-injected) 295.25 d 37.75 32.75 7.97 b 4.39 4.47 a 0.1 ml physiological saline 285.50 cd 41.25 31.75 9.00 ab 4.37 4.66 a 0.1 ml free GEO 314.75 abc 41.00 32.75 6.67 ab 4.37 4.45 bc 0.2 ml free GEO 305.50 bcd 41.00 33.50 9.13 ab 4.54 4.54 ab 0.1 ml nano-encapsulated GEO 356.50 a 44.25 33.75 10.56 a 4.45 4.33 c 0.2 ml nano-encapsulated GEO 343.50 ab 43.75 35.25 9.75 ab 4.49 4.36 c SEM 0.441 1.01 1.45 11.20 0.060 0.033 P -value 0.000 0.064 0.286 0.014 0.273 0.000 a,b,c,d Values within a column with different superscripts differ significantly at P < 0.05. Blood parameter The effects of in-ovo injection of different forms of GEO on the blood parameters of broilers are shown in Table 6 . According to the results, while in-ovo administration did not affect the amount of serum glucose at 7 days and serum cholesterol at 0 and 7 days (P > 0.05), the serum glucose level was affected at zero-day-old chicks (P < 0.05). The highest serum glucose was estimated in the 0.1 free GEO and control treatments, and the lowest was observed in the 0.2 non-encapsulated GEO group. Table 6 Effects of in ovo feeding of garlic essential oil (GEO) on blood parameters of broilers. Treatment Glucose (mg/dl) Cholesterol (mg/dl) 0 d 7 d 0 d 7 d Control (non-injected) 174.5 ab 200.5 421.8 588.5 0.1 ml physiological saline 191.8 a 214.2 422.8 547.0 0.1 ml free GEO 189.5 a 269.5 376.3 581.3 0.2 ml free GEO 135.3 b 192.0 405.0 825.7 0.1 ml nano-encapsulated GEO 171.0 ab 217.0 395.0 516.8 0.2 ml nano-encapsulated GEO 170.0 ab 197.7 402.8 520.3 SEM 10.94 17.24 20.82 38.50 P -value 0.048 0.063 0.630 0.670 a,b Values within a column with different superscripts differ significantly at P < 0.05. Discussion The current study found that the highest percentage of hatch was related to the control treatment and the average weight of chicks at the time of hatching was not affected by the treatments. In another study, Fazli et al. ( 2015 ) reported that in-ovo injection of garlic extract did not seem to affect the hatched chick weight. Furthermore, GEO injected into eggs could not improve body weight in chicks at hatch (Mahjar and Al-Salhie 2022 ). On the other hand, in-ovo administration with 150 µl GEO was found to have a higher hatchability than other treatments and decreased mortality in broiler chickens. Researchers indicated that in-ovo medical plant injection could increase total antioxidant capacity and decrease free radical production, which would lead to improved hatchability (Radwan Nadia et al. 2008 ; Surai et al. 2003 ). It seems that the positive effect of GEO on hatchability may be due to its beneficial role on embryo development through antioxidant and antibacterial properties (Mahjar and Al-Salhie 2022 ). In the present study, BWG, FI and FCR at day 7 and 14 were not affected by any injection treatments. On the impact of in-ovo injection of GEO on chicken performance, however, no research has been found. According to Kumar et al. ( 2022 ), adding nanoencapsulated eugenol to garlic extract in broiler diets had no appreciable impact on FI and FCR of the starter/brooding period (0 to 10 days of age) (Kumar et al. 2022 ). Additionally, feeding laying hens garlic had no noticeable impact on their FI (Adebiyi et al., 2022 ). Other studies investigated the effect of garlic supplementation on broiler performance. The results section states that FI, FCR, and BWG were not significantly influenced by experimental treatment (Onibi et al. 2009 ). The reasons may be related to the gastrointestinal tract and the gut enzyme secretion of broiler chickens which are not fully developed during this period (0 to 14 days of age). The result of the present study shows that there were no significant differences in glycogen levels in both tissues and breast muscle among groups of zero-day chickens. The amount of breast glycogen was improved by the injection of nanoencapsulated GEO, but the amount of liver glycogen was not affected by the treatments. Muscle glycogen levels in poultry meat are closely associated with pH and meat technological quality (Scheffler et al. 2013 ). It is proven that low levels of muscle glycogen indicate a decrease in meat pH (Sugiharto 2022 ).Pre-slaughter stress reduces glycogen stores in poultry muscles and could affect the physical and chemical properties of broiler meat (Sugiharto 2022 ). Also, in a study, Kumar et al. ( 2019 ) reported that garlic supplementation increases the glycogen of broiler muscle. On the other hand, garlic has a variety of bioactive compounds like flavonoids that may increase the glycogen content of the liver with antioxidant properties (Kumar et al. 2019 ). In this trial, nanoencapsulated GEO significantly improved intestinal morphology and microbial population. The villus height and villus height to crypt depth ratios were considerably higher in the 0.1 ml of nanoencapsulated GEO group compared to control group. Also, the in-ovo injection of nanoencapsulated GEO decreased E. coli counts compared to both control groups. Consistent with the results of the present study, Karangiya et al. ( 2016 ) found that the use of 1% garlic in broiler feed significantly increased the villus height. In another study, researchers found that the height, width, and area of the jejunum and duodenum villus increased in broiler chickens supplemented with garlic compared to control (Bahadoran et al. 2016 ). According to Amiri et al. ( 2021 ), a dietary of 100 mg/kg nanoencapsulated GEO showed a higher amount of villus height and villus height to crypt depth ratios in broiler intestine. Interestingly, garlic exhibits higher activity against gram-negative and gram-positive bacteria (Munir 2015 ). In vitro and in vivo, GEO nanoencapsulation with chitosan had significantly higher antibacterial activity against E. coli compared to free form (Amiri et al. 2021 ). It may be suggested that nanoencapsulated GEO has antioxidant properties (Jakubcová et al. 2014 ) in addition to antimicrobial activity (Dávila-Rodrguez et al. 2020). Moreover, it is noteworthy that nanoencapsulation is an improvement method that allows precise and controlled release of bioactive compounds during digestion, which could protect the antibacterial and antioxidant properties of essential oils (Amiri et al. 2021 ). At hatch day, serum glucose increased in the free GEO and control treatments. Although a similar study was not found on the effect of in-ovo injection of GEO on broiler blood parameters, the researchers noticed that adding garlic extract to broiler diets had no significant effect on blood cholesterol levels (Brzóska et al. 2015 ). Furthermore, Parvin et al. ( 2021 ) stated that garlic inclusion in broiler diets significantly reduces blood glucose. In addition, a report indicated that garlic powder in the broiler diet did not significantly change blood cholesterol concentration compared with the control group (Rahimi et al. 2021 ). Glucose levels may be affected by organosulfur compounds like allicin from garlic (Onunkwo et al. 2019 ). Allicin's mode of action is through competing with insulin for its insulin-inactivating sites in the liver, resulting in lower glucose levels (Ala Al Deen 2007). On the other hand, it has been suggested that the compounds in garlic may induce antagonistic activity with insulin and lead to a slight increase in glucose levels (Ala Al Deen 2007). Conclusion According to the results, in-ovo injection of GEO into the yolk sac at day 7 of incubation period reduced the hatching rate. The nanoencapsulated GEO increased breast muscle glycogen, improved intestinal villi, as well as reduced pathogenic populations like E. coli. In general, nanocapsulation with chitosan increased the effectiveness of GEO. Abbreviations GEO, garlic essential oil; EO, essential oils; TPP, sodium triphosphate pentabasic; FI, feed intake; BWG, body weight gain; FCR, feed conversion ratio. Declarations Compliance with Ethical Standards Funding Conflict of Interest The authors declare that they have no conflict of interest. Ethical approval The experiment was conducted according to the animal welfare guidelines at the Veterinary Control and Research Institute of Kerman, Iran (approval number: IR.UK.VETMED.REC 2019-03-05). Consent for publication For this type of study, consent for publication is not required. Author Contribution M.A. conceived and designed the experiment, supervised the project, contributed to data interpretation, and wrote the main manuscript text. N.A. conducted the animal trial, collected samples, and performed the initial data analysis. M.K.B. contributed to statistical analysis and assisted in interpreting the results and manuscript writing. All authors reviewed and approved the final version of the manuscript. Data availability statement Data will be made available on request. 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Scheffler TL, Scheffler JM, Kasten SC, Sosnicki AA, Gerrard DE (2013) High glycolytic potential does not predict low ultimate pH in pork. Meat Sci 95 (1):85-91. https://doi.org/10.1016/j.meatsci.2013.04.013. Sugiharto S (2022) Potential Benefits of Plant-Derived Products on Broiler Meat Characteristics-A Short Review. J. sain peternak. Indones. 17 (1):29-43. https://doi.org/10.1016/j.meatsci.2013.04.013. Surai AP, Surai PF, Steinberg W, Wakeman WG, Speake BK, Sparks NH (2003) Effect of canthaxanthin content of the maternal diet on the antioxidant system of the developing chick. Br Poult Sci 44 (4):612-619. https://doi.org/10.1080/00071660310001616200. Tako E, Ferket PR, Uni Z (2004) Effects of in ovo feeding of carbohydrates and beta-hydroxy-beta-methylbutyrate on the development of chicken intestine. Poult Sci 83 (12):2023-2028. https://doi.org/10.1093/ps/83.12.2023. Yang CM, Cao GT, Ferket PR, Liu TT, Zhou L, Zhang L, Xiao YP, Chen AG (2012) Effects of probiotic, Clostridium butyricum, on growth performance, immune function, and cecal microflora in broiler chickens. Poult Sci 91:2121-2129. https://doi.org/10.3382/ps.2011-02131. Yazdanpanah Goharrizi L, Tasharofi S (2017) Inhibiting Aspergillus flavus growth and aflatoxin decrement in aquatic feed with Satureja hortensis essen-tial oil. Res J Vet Pract 4 (4):71-75. https://doi.org/10.14737/journal.rjvp/2016/4.4.71.75. Additional Declarations No competing interests reported. Cite Share Download PDF Status: Under Review Version 1 posted Editorial decision: Revision requested 07 Sep, 2025 Reviews received at journal 06 Sep, 2025 Reviews received at journal 30 Aug, 2025 Reviewers agreed at journal 26 Aug, 2025 Reviewers agreed at journal 20 Aug, 2025 Reviewers invited by journal 20 Aug, 2025 Editor assigned by journal 16 Jul, 2025 Submission checks completed at journal 16 Jul, 2025 First submitted to journal 14 Jul, 2025 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {\"props\":{\"pageProps\":{\"initialData\":{\"identity\":\"rs-7119473\",\"acceptedTermsAndConditions\":true,\"allowDirectSubmit\":false,\"archivedVersions\":[],\"articleType\":\"Research Article\",\"associatedPublications\":[],\"authors\":[{\"id\":505776055,\"identity\":\"727c2ced-2567-40f8-aaad-bf7d61e35238\",\"order_by\":0,\"name\":\"Negin Amiri\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"Shahid Bahonar University of Kerman\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Negin\",\"middleName\":\"\",\"lastName\":\"Amiri\",\"suffix\":\"\"},{\"id\":505776057,\"identity\":\"8d66b664-5eb8-4805-9bb2-26e98297ebf7\",\"order_by\":1,\"name\":\"Mohsen Afsharmanesh\",\"email\":\"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAAu0lEQVRIiWNgGAWjYFACxgfMQFKOgQfMkyBGC7MBSIsx6VoSG3iIdZZu+2HmzwU199I3nDnA+OEHg0U+QS1mZ5LZpGccK87dcLaBWbKHQcKygaCWA/nHmHnYEnI3nGdgkAb6xYCwLecfM3/m+ZeQbnCegfk3cVpuJDNI87YlJBicbWAj0pYbj9mkefsSDGeeOdhm2WNAlMOSgQ77liDPdyb58I0fFXWEtSABxgYGBpI0jIJRMApGwSjACQDqPTTQl3IjfwAAAABJRU5ErkJggg==\",\"orcid\":\"\",\"institution\":\"Shahid Bahonar University of Kerman\",\"correspondingAuthor\":true,\"prefix\":\"\",\"firstName\":\"Mohsen\",\"middleName\":\"\",\"lastName\":\"Afsharmanesh\",\"suffix\":\"\"},{\"id\":505776060,\"identity\":\"a06e272c-19b1-4531-a627-810087d3a893\",\"order_by\":2,\"name\":\"Mohammad Khajeh Bami\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"Shahid Bahonar University of Kerman\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Mohammad\",\"middleName\":\"Khajeh\",\"lastName\":\"Bami\",\"suffix\":\"\"}],\"badges\":[],\"createdAt\":\"2025-07-14 09:38:11\",\"currentVersionCode\":1,\"declarations\":\"\",\"doi\":\"10.21203/rs.3.rs-7119473/v1\",\"doiUrl\":\"https://doi.org/10.21203/rs.3.rs-7119473/v1\",\"draftVersion\":[],\"editorialEvents\":[],\"editorialNote\":\"\",\"failedWorkflow\":false,\"files\":[{\"id\":90068677,\"identity\":\"0f88d1cf-eebc-439b-abe9-de261d2050d8\",\"added_by\":\"auto\",\"created_at\":\"2025-08-28 06:15:22\",\"extension\":\"pdf\",\"order_by\":0,\"title\":\"\",\"display\":\"\",\"copyAsset\":false,\"role\":\"manuscript-pdf\",\"size\":890126,\"visible\":true,\"origin\":\"\",\"legend\":\"\",\"description\":\"\",\"filename\":\"manuscript.pdf\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-7119473/v1/7722f8a7-78d1-4be7-b4eb-94746809c1ac.pdf\"}],\"financialInterests\":\"No competing interests reported.\",\"formattedTitle\":\"Impact of in ovo feeding of nano-encapsulated garlic essential oil on broiler hatch percentage, performance, intestinal development, and blood parameters\",\"fulltext\":[{\"header\":\"Introduction\",\"content\":\"\\u003cp\\u003eThe poultry meat and egg are widely regarded as suitable sources of nutrients, particularly proteins and during the past several decades, a lot of people have become interested in using poultry raised organically. Additionally, the search for new antimicrobial agents has been prompted by the rise in antibiotic resistance as well as unfavourable side effects of some antibiotic medications (Yazdanpanah Goharrizi and Tasharofi \\u003cspan citationid=\\\"CR32\\\" class=\\\"CitationRef\\\"\\u003e2017\\u003c/span\\u003e). Various plant parts and their essential oils (EO) are recognized for their antibacterial properties and advantageous biological effects (Dehghani et al. \\u003cspan citationid=\\\"CR9\\\" class=\\\"CitationRef\\\"\\u003e2018\\u003c/span\\u003e). One of the most important and beneficial plants used for medicine is garlic (\\u003cem\\u003eAllium sativum\\u003c/em\\u003e). The plant has been used for many centuries to treat intestinal parasites as well as a variety of fungal, viral, and bacterial infections in various nations to prevent health problems (Abd El-latif et al. \\u003cspan citationid=\\\"CR1\\\" class=\\\"CitationRef\\\"\\u003e2013\\u003c/span\\u003e). Diallyl disulfide, allyl methyl trisulfide, allyl (E)-1-propenyl disulfide, and allyl methyl disulfide were all included in the chemical makeup of GEO (Satyal et al. \\u003cspan citationid=\\\"CR25\\\" class=\\\"CitationRef\\\"\\u003e2017\\u003c/span\\u003e).\\u003c/p\\u003e\\u003cp\\u003eContrarily, nanotechnology is a novel technology that can be used to improve food quality and the effectiveness of food production, storage, processing, transportation, traceability, and safety (Mahmoud \\u003cspan citationid=\\\"CR17\\\" class=\\\"CitationRef\\\"\\u003e2012\\u003c/span\\u003e). It is expected that the possibilities will change as a result of nanotechnology. Increased surface area to volume due to nanoprocessing improves absorption and reduces the need for supplements (Jafari et al. \\u003cspan citationid=\\\"CR11\\\" class=\\\"CitationRef\\\"\\u003e2022\\u003c/span\\u003e). Nanoencapsulation, a novel technique to create the potential for a better impression of the covered materials, is the application of nano coating to various materials.\\u003c/p\\u003e\\u003cp\\u003eCombining these two hypotheses shows that using plant EO and nanotechnology yields better results. Therefore, the purpose of this research was to determine the impact of \\u003cem\\u003ein-ovo\\u003c/em\\u003e injection of GEO into the yolk sac on post-hatch broiler chicken performance, intestinal development, hatch percentage, and blood parameters.\\u003c/p\\u003e\"},{\"header\":\"Materials and methods\",\"content\":\"\\u003cp\\u003e\\u003cb\\u003ePreparation of materials\\u003c/b\\u003e\\u003c/p\\u003e\\u003cp\\u003eVolatile oil was distilled from the ground plant material using Clevenger distillation apparatus (Herbal Exir Co., Mashhad, Iran). Ethanol and acetic acid were provided by Merck (NYSE: MRK). Nanoencapsulation were produced using chitosan, derived from crab shell and sodium triphosphate pentabasic (TPP) and provided from Sigma Chemical Company (St. Louis, MO.). Nanoencapsulation of the GEO was performed by ionic gelation according to the procedure of Sawtarie et al. (\\u003cspan citationid=\\\"CR26\\\" class=\\\"CitationRef\\\"\\u003e2017\\u003c/span\\u003e). Chitosan was dissolved at a concentration of 1 mgmL\\u003csup\\u003e\\u0026minus;\\u0026thinsp;1\\u003c/sup\\u003e in 1% (w\\u0026thinsp;=\\u0026thinsp;v) acetic acid and sonicated before the solution became transparent. The dropwise addition of 10mL TPP solution (1 mgmL\\u003csup\\u003e\\u0026minus;\\u0026thinsp;1\\u003c/sup\\u003e) to a 25mL chitosan solution (pH\\u0026thinsp;=\\u0026thinsp;5), under constant stirring at room temperature, produced chitosan-TPP nanoparticles by ionic gelation. For the preparation of chitosan-TPP nanoparticles loaded with herbal EO, 20% (w\\u0026thinsp;=\\u0026thinsp;v) EO were added to the chitosan solution before adding the TPP solution.\\u003c/p\\u003e\\u003cp\\u003e\\u003cb\\u003eDesign and animals\\u003c/b\\u003e\\u003c/p\\u003e\\u003cp\\u003eA total of 360 fertile eggs (flock with 38 week) with mean weight 62\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;2 g (Ross 308) were used in this 5-week trial (3 weeks of incubation and 2 weeks of growing period). Eggs were randomly assigned to 6 treatments with 4 replications of 15 fertile eggs each. The experimental treatments involved: 1) control or non-injected, 2) \\u003cem\\u003ein-ovo\\u003c/em\\u003e injection of 0.1 ml physiological saline or positive control, 3 and 4) \\u003cem\\u003ein-ovo\\u003c/em\\u003e injection of 0.1 and 0.2 ml free GEO, 5 and 6) \\u003cem\\u003ein-ovo\\u003c/em\\u003e injection of 0.1 and 0.2 ml nanoencapsulated GEO.\\u003c/p\\u003e\\u003cp\\u003eSetters with automatic temperature control (37.5\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.1) and rotation of the eggs every 1 hr were maintained at 60% relative humidity until day 18 of incubation, when eggs were transferred to hatcher with 70% relative humidity, the egg rotation was stopped. After hatch, newly-hatched-chicks were kept in pens for 2 weeks. Diet was formulated with UFFDA based on corn and soybean meals (Table\\u0026nbsp;\\u003cspan refid=\\\"Tab1\\\" class=\\\"InternalRef\\\"\\u003e1\\u003c/span\\u003e) and the chicks had ad libitum access to water and feed (formulated according to the standards prescribed in Ross 308 Broiler Nutrition Specification) (Aviagen \\u003cspan citationid=\\\"CR5\\\" class=\\\"CitationRef\\\"\\u003e2014\\u003c/span\\u003e).\\u003c/p\\u003e\\u003cp\\u003e\\u003cdiv class=\\\"gridtable\\\"\\u003e\\u003ctable float=\\\"Yes\\\" id=\\\"Tab1\\\" border=\\\"1\\\"\\u003e\\u003ccaption language=\\\"En\\\"\\u003e\\u003cdiv class=\\\"CaptionNumber\\\"\\u003eTable 1\\u003c/div\\u003e\\u003cdiv class=\\\"CaptionContent\\\"\\u003e\\u003cp\\u003eIngredient and composition of the basal diet.\\u003c/p\\u003e\\u003c/div\\u003e\\u003c/caption\\u003e\\u003ccolgroup cols=\\\"2\\\"\\u003e\\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c1\\\" colnum=\\\"1\\\"\\u003e\\u003c/div\\u003e\\u003cdiv align=\\\"char\\\" char=\\\".\\\" class=\\\"colspec\\\" colname=\\\"c2\\\" colnum=\\\"2\\\"\\u003e\\u003c/div\\u003e\\u003cthead\\u003e\\u003ctr\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003eItem\\u003c/p\\u003e\\u003c/th\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003eAmount\\u003c/p\\u003e\\u003c/th\\u003e\\u003c/tr\\u003e\\u003c/thead\\u003e\\u003ctbody\\u003e\\u003ctr\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003eIngredients (%)\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003eCorn\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003e523.70\\u003c/p\\u003e\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003eSoybean meal\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003e390.00\\u003c/p\\u003e\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003eCorn gluten\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003e25.00\\u003c/p\\u003e\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003eSoybean oil\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003e16.50\\u003c/p\\u003e\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003eCalcium carbonate\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003e12.50\\u003c/p\\u003e\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003eDicalcium phosphate\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003e16.00\\u003c/p\\u003e\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003eSalt\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003e2.50\\u003c/p\\u003e\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003eSodium bicarbonate\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003e2.00\\u003c/p\\u003e\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003eDL-Methionine\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003e3.20\\u003c/p\\u003e\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003eL- Lysine HCL\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003e2.50\\u003c/p\\u003e\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003eL-Threonine\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003e1.00\\u003c/p\\u003e\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003eVitamin and mineral premix\\u003csup\\u003e1\\u003c/sup\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003e5.00\\u003c/p\\u003e\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003ePhytase 10000 (hostazym-p)\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003e0.05\\u003c/p\\u003e\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003eCalculated chemical composition\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003eMetabolisable energy (MJ/kg)\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003e12.20\\u003c/p\\u003e\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003eCrude protein\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003e220.50\\u003c/p\\u003e\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003eMethionine\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003e6.28\\u003c/p\\u003e\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003eMethionine\\u0026thinsp;+\\u0026thinsp;cysteine\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003e9.25\\u003c/p\\u003e\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003eLysine\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003e12.60\\u003c/p\\u003e\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003eThreonine\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003e8.38\\u003c/p\\u003e\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003eTryptophan\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003e2.28\\u003c/p\\u003e\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003eArginine\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003e13.60\\u003c/p\\u003e\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003eCalcium\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003e10.10\\u003c/p\\u003e\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003eAvailable phosphorous\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003e5.03\\u003c/p\\u003e\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003eSodium\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003e1.70\\u003c/p\\u003e\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003eLinoleic acid\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003e19.40\\u003c/p\\u003e\\u003c/td\\u003e\\u003c/tr\\u003e\\u003c/tbody\\u003e\\u003c/colgroup\\u003e\\u003ctfoot\\u003e\\u003ctr\\u003e\\u003ctd colspan=\\\"2\\\"\\u003e\\u003csup\\u003e1\\u003c/sup\\u003e Provided per kg of diet: retinyl acetate, 10000 IU; cholecalciferol, 4500 IU; DL-α-tocopheryl acetate, 65 IU; menadione, 3 mg; thiamin, 2.5 mg; Riboflavin, 5.2 mg; Niacin, 35 mg; Pantothenic Acid, 18 mg; pyridoxine, 3.2 mg; Biotin, 0.18 mg; Folic acid, 1.6 mg; cyanocobalamin, 0.017 mg; choline choloride, 800 mg and antioxidant, 2.5 mg. Provided per kg of diet: copper, 16 mg; iodine, 1.25 mg; iron,20 mg; manganese,120 mg; selenium, 0.30 mg and zinc, 100 mg.\\u003c/td\\u003e\\u003c/tr\\u003e\\u003c/tfoot\\u003e\\u003c/table\\u003e\\u003c/div\\u003e\\u003c/p\\u003e\\u003cp\\u003e\\u003cb\\u003eIn-ovo injection procedure\\u003c/b\\u003e\\u003c/p\\u003e\\u003cp\\u003eThe injection site was disinfected with 96% alcohol, and then, 0.1 and 0.2 ml of each solution was injected into the yolk sac at day 7 of incubation period, using a 23-gauge needle with depth of 28 mm from the broad end of the egg. Then, the holes were sealed using melt paraffin.\\u003c/p\\u003e\\u003cp\\u003e\\u003cb\\u003eData collection\\u003c/b\\u003e\\u003c/p\\u003e\\u003cp\\u003eThe hatch percentage and chicken weight on the first day for every replicate was recorded and then experimental period lasted 14 days. The feed amounts offered and refused were measured at days 7 and 14 for each pen to calculate feed intake (FI). Also, chicks were weighed at above-mentioned days to calculate body weight gain (BWG) changes. By having these two measurements, feed conversion ratio (FCR) was calculated as feed intake consumed per unit of weight gained.\\u003c/p\\u003e\\u003cp\\u003eAt days of hatch and 7, two chicks from every replicate were slaughtered to measure glycogen content of liver and breast and jejunum morphology. Within 1 min after the birds were killed, liver and breast were taken in plastic bags and frozen in -20\\u0026deg;C until the start of procedure. Afterwards, 0.2 g samples of liver and breast were added to 1 ml of 8% perchloric acid, homogenized (in ice) for 1 min and centrifuged at 14,000 \\u0026times; g at 4\\u0026deg;C for 30 min. The supernatant was removed, and 1 ml petroleum ether (60\\u0026ndash;80\\u0026deg;C) was added. After being mixed, the petroleum ether fraction was removed, and samples from the bottom layer were transferred to a new tube for glycogen determination.\\u003c/p\\u003e\\u003cp\\u003eFor histopathologic analysis, 2-cm tissue samples from the middle of jejunum were obtained and fixed in 10% buffered formalin (100 ml of 40% formaldehyde, 4 g phosphate, 6.5 g dibasic sodium phosphate and 900 ml of distilled water) for 24 hr, and then, the 10% buffered formalin was renewed. Tissues were dehydrated by transferring through a series of alcohols with increasing concentrations, placed into xylol and embedded in paraffin. A microtome was used to make five cuts that were 5 \\u0026micro;m. The cuts were stained with haematoxylin\\u0026ndash;eosin. The values were measured using a light microscope. Measurements of villus height and width and crypt depth were determined at a magnification of 10X. A minimum of five measurements per slide were made for each parameter and averaged into one value (Tako et al. \\u003cspan citationid=\\\"CR30\\\" class=\\\"CitationRef\\\"\\u003e2004\\u003c/span\\u003e).\\u003c/p\\u003e\\u003cp\\u003eThe contents of ileum (10 cm anterior to the junction with caecum and rectum) were collected from 60 birds and put into sterilized vials, and then went for counting the microbial population such as \\u003cem\\u003eE. coli\\u003c/em\\u003e and \\u003cem\\u003eLactobacilli\\u003c/em\\u003e which transported immediately to the laboratory. Population of \\u003cem\\u003eE. coli\\u003c/em\\u003e and \\u003cem\\u003eLactobacilli\\u003c/em\\u003e were estimated as CFU g\\u003csup\\u003e\\u0026minus;\\u0026thinsp;1\\u003c/sup\\u003e using subsequent dilution method. Bacterial counts were performed and cultured using MacConkey agar (at 37\\u0026deg;C in an incubator for 1 d) for \\u003cem\\u003eE. coli\\u003c/em\\u003e and De Man, Rogosa and Sharpe (at 37\\u0026deg;C in an incubator for 2 d) for \\u003cem\\u003eLactobacilli\\u003c/em\\u003e (Yang et al. \\u003cspan citationid=\\\"CR31\\\" class=\\\"CitationRef\\\"\\u003e2012\\u003c/span\\u003e).\\u003c/p\\u003e\\u003cp\\u003eBlood samples were obtained from chicks mentioned above and drawn into tubes. After coagulation, the serum samples were centrifuged at 4000 rpm for 10 min, and analysis of serum glucose and cholesterol contents was carried out by enzyme procedure of CHOD-PAP and biochemistry kit (Asan Pharm. Co., Korea) (Richmond \\u003cspan citationid=\\\"CR24\\\" class=\\\"CitationRef\\\"\\u003e1973\\u003c/span\\u003e).\\u003c/p\\u003e\\u003cdiv id=\\\"Sec3\\\" class=\\\"Section2\\\"\\u003e\\u003ch2\\u003eStatistical analysis\\u003c/h2\\u003e\\u003cp\\u003eAll data were subject to analysis of variance following the GLM procedure in a completely randomized design using the SAS software program. The difference among treatments was compared using the Duncan\\u0026rsquo;s multiple range test. The treatment effect was considered significant at a probability value below 0.05.\\u003c/p\\u003e\\u003c/div\\u003e\"},{\"header\":\"Results\",\"content\":\"\\u003cp\\u003e\\u003cb\\u003eHatchability and body weight\\u003c/b\\u003e\\u003c/p\\u003e\\u003cp\\u003eThe effects of injecting several types of GEO in ovo on hatchability and chick body weight at hatching are shown in Table\\u0026nbsp;\\u003cspan refid=\\\"Tab2\\\" class=\\\"InternalRef\\\"\\u003e2\\u003c/span\\u003e. The results showed that \\u003cem\\u003ein-ovo\\u003c/em\\u003e administration of GEO had a substantial impact on hatchability but experimental treatments had no significant impact on the body weight of the chicks (P\\u0026thinsp;\\u0026gt;\\u0026thinsp;0.05). The free GEO groups (0.1 and 0.2 ml) had the lowest hatchability, whereas the control treatment (without injection) had the highest. The lack of a noticeable difference between the treatments containing encapsulated GEO and the control group is interesting (without injection).\\u003c/p\\u003e\\u003cp\\u003e\\u003cdiv class=\\\"gridtable\\\"\\u003e\\u003ctable float=\\\"Yes\\\" id=\\\"Tab2\\\" border=\\\"1\\\"\\u003e\\u003ccaption language=\\\"En\\\"\\u003e\\u003cdiv class=\\\"CaptionNumber\\\"\\u003eTable 2\\u003c/div\\u003e\\u003cdiv class=\\\"CaptionContent\\\"\\u003e\\u003cp\\u003eEffects of in ovo feeding of garlic essential oil (GEO) on hatchability and hatched chick weight.\\u003c/p\\u003e\\u003c/div\\u003e\\u003c/caption\\u003e\\u003ccolgroup cols=\\\"3\\\"\\u003e\\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c1\\\" colnum=\\\"1\\\"\\u003e\\u003c/div\\u003e\\u003cdiv align=\\\"char\\\" char=\\\".\\\" class=\\\"colspec\\\" colname=\\\"c2\\\" colnum=\\\"2\\\"\\u003e\\u003c/div\\u003e\\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c3\\\" colnum=\\\"3\\\"\\u003e\\u003c/div\\u003e\\u003cthead\\u003e\\u003ctr\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003eTreatment\\u003c/p\\u003e\\u003c/th\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003eWeight (g)\\u003c/p\\u003e\\u003c/th\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c3\\\"\\u003e\\u003cp\\u003eHatch (%)\\u003c/p\\u003e\\u003c/th\\u003e\\u003c/tr\\u003e\\u003c/thead\\u003e\\u003ctbody\\u003e\\u003ctr\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003eControl (non-injected)\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003e43.31\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e\\u003cp\\u003e89.50\\u003csup\\u003ea\\u003c/sup\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003e0.1 ml physiological saline\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003e43.93\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e\\u003cp\\u003e82.75\\u003csup\\u003ebc\\u003c/sup\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003e0.1 ml free GEO\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003e43.71\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e\\u003cp\\u003e79.50\\u003csup\\u003ec\\u003c/sup\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003e0.2 ml free GEO\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003e43.80\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e\\u003cp\\u003e81.00\\u003csup\\u003ec\\u003c/sup\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003e0.1 ml nano-encapsulated GEO\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003e44.06\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e\\u003cp\\u003e88.00\\u003csup\\u003eab\\u003c/sup\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003e0.2 ml nano-encapsulated GEO\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003e43.48\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e\\u003cp\\u003e87.50\\u003csup\\u003eab\\u003c/sup\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003eSEM\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003e0.182\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e\\u003cp\\u003e1.26\\u003c/p\\u003e\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003e\\u003cem\\u003eP\\u003c/em\\u003e-value\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003e0.085\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e\\u003cp\\u003e0.000\\u003c/p\\u003e\\u003c/td\\u003e\\u003c/tr\\u003e\\u003c/tbody\\u003e\\u003c/colgroup\\u003e\\u003ctfoot\\u003e\\u003ctr\\u003e\\u003ctd colspan=\\\"3\\\"\\u003e\\u003csup\\u003ea,b,c\\u003c/sup\\u003e Values within a column with different superscripts differ significantly at P\\u0026thinsp;\\u0026lt;\\u0026thinsp;0.05.\\u003c/td\\u003e\\u003c/tr\\u003e\\u003c/tfoot\\u003e\\u003c/table\\u003e\\u003c/div\\u003e\\u003c/p\\u003e\\u003cp\\u003e\\u003cb\\u003ePerformance\\u003c/b\\u003e\\u003c/p\\u003e\\u003cp\\u003eTable\\u0026nbsp;\\u003cspan refid=\\\"Tab3\\\" class=\\\"InternalRef\\\"\\u003e3\\u003c/span\\u003e shows the impact of \\u003cem\\u003ein-ovo\\u003c/em\\u003e feeding different types of GEO on the performance of broiler chickens. The findings show that experimental treatments had no effect on BWG, FI, or FCR (P\\u0026thinsp;\\u0026gt;\\u0026thinsp;0.05).\\u003c/p\\u003e\\u003cp\\u003e\\u003cdiv class=\\\"gridtable\\\"\\u003e\\u003ctable float=\\\"Yes\\\" id=\\\"Tab3\\\" border=\\\"1\\\"\\u003e\\u003ccaption language=\\\"En\\\"\\u003e\\u003cdiv class=\\\"CaptionNumber\\\"\\u003eTable 3\\u003c/div\\u003e\\u003cdiv class=\\\"CaptionContent\\\"\\u003e\\u003cp\\u003eEffects of in ovo feeding of garlic essential oil (GEO) on growth performance of broilers (1 to 14 d).\\u003c/p\\u003e\\u003c/div\\u003e\\u003c/caption\\u003e\\u003ccolgroup cols=\\\"7\\\"\\u003e\\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c1\\\" colnum=\\\"1\\\"\\u003e\\u003c/div\\u003e\\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c2\\\" colnum=\\\"2\\\"\\u003e\\u003c/div\\u003e\\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c3\\\" colnum=\\\"3\\\"\\u003e\\u003c/div\\u003e\\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c4\\\" colnum=\\\"4\\\"\\u003e\\u003c/div\\u003e\\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c5\\\" colnum=\\\"5\\\"\\u003e\\u003c/div\\u003e\\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c6\\\" colnum=\\\"6\\\"\\u003e\\u003c/div\\u003e\\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c7\\\" colnum=\\\"7\\\"\\u003e\\u003c/div\\u003e\\u003cthead\\u003e\\u003ctr\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003eTreatment\\u003c/p\\u003e\\u003c/th\\u003e\\u003cth align=\\\"left\\\" colspan=\\\"2\\\" nameend=\\\"c3\\\" namest=\\\"c2\\\"\\u003e\\u003cp\\u003eBody weight gain\\u003c/p\\u003e\\u003cp\\u003e(g/b/d)\\u003c/p\\u003e\\u003c/th\\u003e\\u003cth align=\\\"left\\\" colspan=\\\"2\\\" nameend=\\\"c5\\\" namest=\\\"c4\\\"\\u003e\\u003cp\\u003eFeed intake\\u003c/p\\u003e\\u003cp\\u003e(g/b/d)\\u003c/p\\u003e\\u003c/th\\u003e\\u003cth align=\\\"left\\\" colspan=\\\"2\\\" nameend=\\\"c7\\\" namest=\\\"c6\\\"\\u003e\\u003cp\\u003eFeed conversion ratio (g/g)\\u003c/p\\u003e\\u003c/th\\u003e\\u003c/tr\\u003e\\u003c/thead\\u003e\\u003ctbody\\u003e\\u003ctr\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003e1\\u0026ndash;7 d\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e\\u003cp\\u003e8\\u0026ndash;14 d\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e\\u003cp\\u003e1\\u0026ndash;7 d\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e\\u003cp\\u003e8\\u0026ndash;14 d\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e\\u003cp\\u003e1\\u0026ndash;7 d\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e\\u003cp\\u003e8\\u0026ndash;14 d\\u003c/p\\u003e\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003eControl (non-injected)\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003e147.1\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e\\u003cp\\u003e344.4\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e\\u003cp\\u003e149.3\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e\\u003cp\\u003e442.3\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e\\u003cp\\u003e1.01\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e\\u003cp\\u003e1.28\\u003c/p\\u003e\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003e0.1 ml physiological saline\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003e147.2\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e\\u003cp\\u003e337.6\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e\\u003cp\\u003e147.9\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e\\u003cp\\u003e440.9\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e\\u003cp\\u003e1.00\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e\\u003cp\\u003e1.30\\u003c/p\\u003e\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003e0.1 ml free GEO\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003e147.7\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e\\u003cp\\u003e339.1\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e\\u003cp\\u003e148.2\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e\\u003cp\\u003e441.2\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e\\u003cp\\u003e1.00\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e\\u003cp\\u003e1.30\\u003c/p\\u003e\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003e0.2 ml free GEO\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003e147.9\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e\\u003cp\\u003e338.4\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e\\u003cp\\u003e150.6\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e\\u003cp\\u003e443.6\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e\\u003cp\\u003e1.01\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e\\u003cp\\u003e1.31\\u003c/p\\u003e\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003e0.1 ml nano-encapsulated GEO\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003e151.0\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e\\u003cp\\u003e348.5\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e\\u003cp\\u003e148.2\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e\\u003cp\\u003e440.5\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e\\u003cp\\u003e0.982\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e\\u003cp\\u003e1.26\\u003c/p\\u003e\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003e0.2 ml nano-encapsulated GEO\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003e150.2\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e\\u003cp\\u003e348.2\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e\\u003cp\\u003e148.8\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e\\u003cp\\u003e440.8\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e\\u003cp\\u003e0.990\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e\\u003cp\\u003e1.26\\u003c/p\\u003e\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003eSEM\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003e1.50\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e\\u003cp\\u003e3.11\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e\\u003cp\\u003e1.35\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e\\u003cp\\u003e1.36\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e\\u003cp\\u003e0.014\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e\\u003cp\\u003e0.012\\u003c/p\\u003e\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003e\\u003cem\\u003eP\\u003c/em\\u003e-value\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003e0.344\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e\\u003cp\\u003e0.064\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e\\u003cp\\u003e0.739\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e\\u003cp\\u003e0.583\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e\\u003cp\\u003e0.496\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e\\u003cp\\u003e0.067\\u003c/p\\u003e\\u003c/td\\u003e\\u003c/tr\\u003e\\u003c/tbody\\u003e\\u003c/colgroup\\u003e\\u003c/table\\u003e\\u003c/div\\u003e\\u003c/p\\u003e\\u003cp\\u003e\\u003cb\\u003eGlycogen content of liver and pectoral muscle\\u003c/b\\u003e\\u003c/p\\u003e\\u003cp\\u003eThe concentrations of glycogen in the breasts of 7-day-old chicks were significantly influenced (P\\u0026thinsp;\\u0026lt;\\u0026thinsp;0.05) (Table\\u0026nbsp;\\u003cspan refid=\\\"Tab4\\\" class=\\\"InternalRef\\\"\\u003e4\\u003c/span\\u003e). So, the level of breast glycogen was highest in the 0.1 nanoencapsulated GEO group, whereas the lowest level was observed in the control group. Liver glycogen concentration at hatch and 7 days old were not significantly different between treatments.\\u003c/p\\u003e\\u003cp\\u003e\\u003cdiv class=\\\"gridtable\\\"\\u003e\\u003ctable float=\\\"Yes\\\" id=\\\"Tab4\\\" border=\\\"1\\\"\\u003e\\u003ccaption language=\\\"En\\\"\\u003e\\u003cdiv class=\\\"CaptionNumber\\\"\\u003eTable 4\\u003c/div\\u003e\\u003cdiv class=\\\"CaptionContent\\\"\\u003e\\u003cp\\u003eEffects of in ovo feeding of garlic essential oil (GEO) on glycogen levels in both liver tissue and breast muscle of broilers.\\u003c/p\\u003e\\u003c/div\\u003e\\u003c/caption\\u003e\\u003ccolgroup cols=\\\"6\\\"\\u003e\\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c1\\\" colnum=\\\"1\\\"\\u003e\\u003c/div\\u003e\\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c2\\\" colnum=\\\"2\\\"\\u003e\\u003c/div\\u003e\\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c3\\\" colnum=\\\"3\\\"\\u003e\\u003c/div\\u003e\\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c4\\\" colnum=\\\"4\\\"\\u003e\\u003c/div\\u003e\\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c5\\\" colnum=\\\"5\\\"\\u003e\\u003c/div\\u003e\\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c6\\\" colnum=\\\"6\\\"\\u003e\\u003c/div\\u003e\\u003cthead\\u003e\\u003ctr\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003eTreatment\\u003c/p\\u003e\\u003c/th\\u003e\\u003cth align=\\\"left\\\" colspan=\\\"2\\\" nameend=\\\"c3\\\" namest=\\\"c2\\\"\\u003e\\u003cp\\u003eBreast glycogen\\u003c/p\\u003e\\u003c/th\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c4\\\"\\u003e\\u0026nbsp;\\u003c/th\\u003e\\u003cth align=\\\"left\\\" colspan=\\\"2\\\" nameend=\\\"c6\\\" namest=\\\"c5\\\"\\u003e\\u003cp\\u003eLiver glycogen\\u003c/p\\u003e\\u003c/th\\u003e\\u003c/tr\\u003e\\u003c/thead\\u003e\\u003ctbody\\u003e\\u003ctr\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003e0 d\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e\\u003cp\\u003e7 d\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e\\u003cp\\u003e0 d\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e\\u003cp\\u003e7 d\\u003c/p\\u003e\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003eControl (non-injected)\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003e0.153\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e\\u003cp\\u003e0.280\\u003csup\\u003eb\\u003c/sup\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e\\u003cp\\u003e0.136\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e\\u003cp\\u003e1.04\\u003c/p\\u003e\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003e0.1 ml physiological saline\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003e0.214\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e\\u003cp\\u003e0.302\\u003csup\\u003eab\\u003c/sup\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e\\u003cp\\u003e0.122\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e\\u003cp\\u003e0.953\\u003c/p\\u003e\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003e0.1 ml free GEO\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003e0.214\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e\\u003cp\\u003e0.292\\u003csup\\u003eab\\u003c/sup\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e\\u003cp\\u003e0.190\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e\\u003cp\\u003e0.946\\u003c/p\\u003e\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003e0.2 ml free GEO\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003e0.235\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e\\u003cp\\u003e0.359\\u003csup\\u003eab\\u003c/sup\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e\\u003cp\\u003e0.173\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e\\u003cp\\u003e0.989\\u003c/p\\u003e\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003e0.1 ml nano-encapsulated GEO\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003e0.237\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e\\u003cp\\u003e0.401\\u003csup\\u003ea\\u003c/sup\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e\\u003cp\\u003e0.174\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e\\u003cp\\u003e1.06\\u003c/p\\u003e\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003e0.2 ml nano-encapsulated GEO\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003e0.235\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e\\u003cp\\u003e0.391\\u003csup\\u003eab\\u003c/sup\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e\\u003cp\\u003e0.165\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e\\u003cp\\u003e1.14\\u003c/p\\u003e\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003eSEM\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003e0.027\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e\\u003cp\\u003e0.025\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e\\u003cp\\u003e0.024\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e\\u003cp\\u003e0.053\\u003c/p\\u003e\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003e\\u003cem\\u003eP\\u003c/em\\u003e-value\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003e0.284\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e\\u003cp\\u003e0.009\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e\\u003cp\\u003e0.383\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e\\u003cp\\u003e0.134\\u003c/p\\u003e\\u003c/td\\u003e\\u003c/tr\\u003e\\u003c/tbody\\u003e\\u003c/colgroup\\u003e\\u003ctfoot\\u003e\\u003ctr\\u003e\\u003ctd colspan=\\\"6\\\"\\u003e\\u003csup\\u003ea,b\\u003c/sup\\u003e Values within a column with different superscripts differ significantly at P\\u0026thinsp;\\u0026lt;\\u0026thinsp;0.05.\\u003c/td\\u003e\\u003c/tr\\u003e\\u003c/tfoot\\u003e\\u003c/table\\u003e\\u003c/div\\u003e\\u003c/p\\u003e\\u003cp\\u003e\\u003cb\\u003eJejunum morphology and microbial population\\u003c/b\\u003e\\u003c/p\\u003e\\u003cp\\u003eThe effect of \\u003cem\\u003ein-ovo\\u003c/em\\u003e feeding of different forms of GEO on intestinal morphology and the microbial population of broiler chickens is presented in Table\\u0026nbsp;\\u003cspan refid=\\\"Tab5\\\" class=\\\"InternalRef\\\"\\u003e5\\u003c/span\\u003e. Based on the results, the villus width, crypt depth, and \\u003cem\\u003elactobacillus\\u003c/em\\u003e bacterial population were not influenced by experimental groups (P\\u0026thinsp;\\u0026gt;\\u0026thinsp;0.05). However, villus height, villus height to crypt depth ratios, and \\u003cem\\u003eE. coli\\u003c/em\\u003e bacterial population were significantly affected (P\\u0026thinsp;\\u0026lt;\\u0026thinsp;0.05). The villus height and villus height to crypt depth ratios were considerably higher in the 0.1 ml of nanoencapsulated GEO group compared to control group (P\\u0026thinsp;\\u0026lt;\\u0026thinsp;0.05). Also, the \\u003cem\\u003ein-ovo\\u003c/em\\u003e injection of nanoencapsulated GEO decreased \\u003cem\\u003eE. coli\\u003c/em\\u003e counts compared to both control groups (P\\u0026thinsp;\\u0026lt;\\u0026thinsp;0.05).\\u003c/p\\u003e\\u003cp\\u003e\\u003cdiv class=\\\"gridtable\\\"\\u003e\\u003ctable float=\\\"Yes\\\" id=\\\"Tab5\\\" border=\\\"1\\\"\\u003e\\u003ccaption language=\\\"En\\\"\\u003e\\u003cdiv class=\\\"CaptionNumber\\\"\\u003eTable 5\\u003c/div\\u003e\\u003cdiv class=\\\"CaptionContent\\\"\\u003e\\u003cp\\u003eEffects of in ovo feeding of garlic essential oil (GEO) on intestinal morphology and microbial population of broilers.\\u003c/p\\u003e\\u003c/div\\u003e\\u003c/caption\\u003e\\u003ccolgroup cols=\\\"7\\\"\\u003e\\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c1\\\" colnum=\\\"1\\\"\\u003e\\u003c/div\\u003e\\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c2\\\" colnum=\\\"2\\\"\\u003e\\u003c/div\\u003e\\u003cdiv align=\\\"char\\\" char=\\\".\\\" class=\\\"colspec\\\" colname=\\\"c3\\\" colnum=\\\"3\\\"\\u003e\\u003c/div\\u003e\\u003cdiv align=\\\"char\\\" char=\\\".\\\" class=\\\"colspec\\\" colname=\\\"c4\\\" colnum=\\\"4\\\"\\u003e\\u003c/div\\u003e\\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c5\\\" colnum=\\\"5\\\"\\u003e\\u003c/div\\u003e\\u003cdiv align=\\\"char\\\" char=\\\".\\\" class=\\\"colspec\\\" colname=\\\"c6\\\" colnum=\\\"6\\\"\\u003e\\u003c/div\\u003e\\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c7\\\" colnum=\\\"7\\\"\\u003e\\u003c/div\\u003e\\u003cthead\\u003e\\u003ctr\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003eTreatment\\u003c/p\\u003e\\u003c/th\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003eVillus height\\u003c/p\\u003e\\u003cp\\u003e\\u0026micro;m))\\u003c/p\\u003e\\u003c/th\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c3\\\"\\u003e\\u003cp\\u003eVillus width\\u003c/p\\u003e\\u003cp\\u003e(\\u0026micro;m)\\u003c/p\\u003e\\u003c/th\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c4\\\"\\u003e\\u003cp\\u003eCrypt depth\\u003c/p\\u003e\\u003cp\\u003e\\u0026micro;m))\\u003c/p\\u003e\\u003c/th\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c5\\\"\\u003e\\u003cp\\u003eVillus height/crypt depth\\u003c/p\\u003e\\u003c/th\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c6\\\"\\u003e\\u003cp\\u003eLactobacillus\\u003c/p\\u003e\\u003c/th\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c7\\\"\\u003e\\u003cp\\u003e\\u003cem\\u003eE.coli\\u003c/em\\u003e\\u003c/p\\u003e\\u003c/th\\u003e\\u003c/tr\\u003e\\u003c/thead\\u003e\\u003ctbody\\u003e\\u003ctr\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003eControl (non-injected)\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003e295.25\\u003csup\\u003ed\\u003c/sup\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c3\\\"\\u003e\\u003cp\\u003e37.75\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c4\\\"\\u003e\\u003cp\\u003e32.75\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e\\u003cp\\u003e7.97\\u003csup\\u003eb\\u003c/sup\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c6\\\"\\u003e\\u003cp\\u003e4.39\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e\\u003cp\\u003e4.47\\u003csup\\u003ea\\u003c/sup\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003e0.1 ml physiological saline\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003e285.50\\u003csup\\u003ecd\\u003c/sup\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c3\\\"\\u003e\\u003cp\\u003e41.25\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c4\\\"\\u003e\\u003cp\\u003e31.75\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e\\u003cp\\u003e9.00\\u003csup\\u003eab\\u003c/sup\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c6\\\"\\u003e\\u003cp\\u003e4.37\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e\\u003cp\\u003e4.66\\u003csup\\u003ea\\u003c/sup\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003e0.1 ml free GEO\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003e314.75\\u003csup\\u003eabc\\u003c/sup\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c3\\\"\\u003e\\u003cp\\u003e41.00\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c4\\\"\\u003e\\u003cp\\u003e32.75\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e\\u003cp\\u003e6.67\\u003csup\\u003eab\\u003c/sup\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c6\\\"\\u003e\\u003cp\\u003e4.37\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e\\u003cp\\u003e4.45\\u003csup\\u003ebc\\u003c/sup\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003e0.2 ml free GEO\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003e305.50\\u003csup\\u003ebcd\\u003c/sup\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c3\\\"\\u003e\\u003cp\\u003e41.00\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c4\\\"\\u003e\\u003cp\\u003e33.50\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e\\u003cp\\u003e9.13\\u003csup\\u003eab\\u003c/sup\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c6\\\"\\u003e\\u003cp\\u003e4.54\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e\\u003cp\\u003e4.54\\u003csup\\u003eab\\u003c/sup\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003e0.1 ml nano-encapsulated GEO\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003e356.50\\u003csup\\u003ea\\u003c/sup\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c3\\\"\\u003e\\u003cp\\u003e44.25\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c4\\\"\\u003e\\u003cp\\u003e33.75\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e\\u003cp\\u003e10.56\\u003csup\\u003ea\\u003c/sup\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c6\\\"\\u003e\\u003cp\\u003e4.45\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e\\u003cp\\u003e4.33\\u003csup\\u003ec\\u003c/sup\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003e0.2 ml nano-encapsulated GEO\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003e343.50\\u003csup\\u003eab\\u003c/sup\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c3\\\"\\u003e\\u003cp\\u003e43.75\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c4\\\"\\u003e\\u003cp\\u003e35.25\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e\\u003cp\\u003e9.75\\u003csup\\u003eab\\u003c/sup\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c6\\\"\\u003e\\u003cp\\u003e4.49\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e\\u003cp\\u003e4.36\\u003csup\\u003ec\\u003c/sup\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003eSEM\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003e0.441\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c3\\\"\\u003e\\u003cp\\u003e1.01\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c4\\\"\\u003e\\u003cp\\u003e1.45\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e\\u003cp\\u003e11.20\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c6\\\"\\u003e\\u003cp\\u003e0.060\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e\\u003cp\\u003e0.033\\u003c/p\\u003e\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003e\\u003cem\\u003eP\\u003c/em\\u003e-value\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003e0.000\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c3\\\"\\u003e\\u003cp\\u003e0.064\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c4\\\"\\u003e\\u003cp\\u003e0.286\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e\\u003cp\\u003e0.014\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c6\\\"\\u003e\\u003cp\\u003e0.273\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e\\u003cp\\u003e0.000\\u003c/p\\u003e\\u003c/td\\u003e\\u003c/tr\\u003e\\u003c/tbody\\u003e\\u003c/colgroup\\u003e\\u003ctfoot\\u003e\\u003ctr\\u003e\\u003ctd colspan=\\\"7\\\"\\u003e\\u003csup\\u003ea,b,c,d\\u003c/sup\\u003e Values within a column with different superscripts differ significantly at P\\u0026thinsp;\\u0026lt;\\u0026thinsp;0.05.\\u003c/td\\u003e\\u003c/tr\\u003e\\u003c/tfoot\\u003e\\u003c/table\\u003e\\u003c/div\\u003e\\u003c/p\\u003e\\u003cp\\u003e\\u003cb\\u003eBlood parameter\\u003c/b\\u003e\\u003c/p\\u003e\\u003cp\\u003eThe effects of \\u003cem\\u003ein-ovo\\u003c/em\\u003e injection of different forms of GEO on the blood parameters of broilers are shown in Table\\u0026nbsp;\\u003cspan refid=\\\"Tab6\\\" class=\\\"InternalRef\\\"\\u003e6\\u003c/span\\u003e. According to the results, while \\u003cem\\u003ein-ovo\\u003c/em\\u003e administration did not affect the amount of serum glucose at 7 days and serum cholesterol at 0 and 7 days (P\\u0026thinsp;\\u0026gt;\\u0026thinsp;0.05), the serum glucose level was affected at zero-day-old chicks (P\\u0026thinsp;\\u0026lt;\\u0026thinsp;0.05). The highest serum glucose was estimated in the 0.1 free GEO and control treatments, and the lowest was observed in the 0.2 non-encapsulated GEO group.\\u003c/p\\u003e\\u003cp\\u003e\\u003cdiv class=\\\"gridtable\\\"\\u003e\\u003ctable float=\\\"Yes\\\" id=\\\"Tab6\\\" border=\\\"1\\\"\\u003e\\u003ccaption language=\\\"En\\\"\\u003e\\u003cdiv class=\\\"CaptionNumber\\\"\\u003eTable 6\\u003c/div\\u003e\\u003cdiv class=\\\"CaptionContent\\\"\\u003e\\u003cp\\u003eEffects of in ovo feeding of garlic essential oil (GEO) on blood parameters of broilers.\\u003c/p\\u003e\\u003c/div\\u003e\\u003c/caption\\u003e\\u003ccolgroup cols=\\\"6\\\"\\u003e\\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c1\\\" colnum=\\\"1\\\"\\u003e\\u003c/div\\u003e\\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c2\\\" colnum=\\\"2\\\"\\u003e\\u003c/div\\u003e\\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c3\\\" colnum=\\\"3\\\"\\u003e\\u003c/div\\u003e\\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c4\\\" colnum=\\\"4\\\"\\u003e\\u003c/div\\u003e\\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c5\\\" colnum=\\\"5\\\"\\u003e\\u003c/div\\u003e\\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c6\\\" colnum=\\\"6\\\"\\u003e\\u003c/div\\u003e\\u003cthead\\u003e\\u003ctr\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003eTreatment\\u003c/p\\u003e\\u003c/th\\u003e\\u003cth align=\\\"left\\\" colspan=\\\"2\\\" nameend=\\\"c3\\\" namest=\\\"c2\\\"\\u003e\\u003cp\\u003eGlucose (mg/dl)\\u003c/p\\u003e\\u003c/th\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c4\\\"\\u003e\\u0026nbsp;\\u003c/th\\u003e\\u003cth align=\\\"left\\\" colspan=\\\"2\\\" nameend=\\\"c6\\\" namest=\\\"c5\\\"\\u003e\\u003cp\\u003eCholesterol (mg/dl)\\u003c/p\\u003e\\u003c/th\\u003e\\u003c/tr\\u003e\\u003c/thead\\u003e\\u003ctbody\\u003e\\u003ctr\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003e0 d\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e\\u003cp\\u003e7 d\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e\\u003cp\\u003e0 d\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e\\u003cp\\u003e7 d\\u003c/p\\u003e\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003eControl (non-injected)\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003e174.5\\u003csup\\u003eab\\u003c/sup\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e\\u003cp\\u003e200.5\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e\\u003cp\\u003e421.8\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e\\u003cp\\u003e588.5\\u003c/p\\u003e\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003e0.1 ml physiological saline\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003e191.8\\u003csup\\u003ea\\u003c/sup\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e\\u003cp\\u003e214.2\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e\\u003cp\\u003e422.8\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e\\u003cp\\u003e547.0\\u003c/p\\u003e\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003e0.1 ml free GEO\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003e189.5\\u003csup\\u003ea\\u003c/sup\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e\\u003cp\\u003e269.5\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e\\u003cp\\u003e376.3\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e\\u003cp\\u003e581.3\\u003c/p\\u003e\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003e0.2 ml free GEO\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003e135.3\\u003csup\\u003eb\\u003c/sup\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e\\u003cp\\u003e192.0\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e\\u003cp\\u003e405.0\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e\\u003cp\\u003e825.7\\u003c/p\\u003e\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003e0.1 ml nano-encapsulated GEO\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003e171.0\\u003csup\\u003eab\\u003c/sup\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e\\u003cp\\u003e217.0\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e\\u003cp\\u003e395.0\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e\\u003cp\\u003e516.8\\u003c/p\\u003e\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003e0.2 ml nano-encapsulated GEO\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003e170.0\\u003csup\\u003eab\\u003c/sup\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e\\u003cp\\u003e197.7\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e\\u003cp\\u003e402.8\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e\\u003cp\\u003e520.3\\u003c/p\\u003e\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003eSEM\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003e10.94\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e\\u003cp\\u003e17.24\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e\\u003cp\\u003e20.82\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e\\u003cp\\u003e38.50\\u003c/p\\u003e\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003e\\u003cem\\u003eP\\u003c/em\\u003e-value\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003e0.048\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e\\u003cp\\u003e0.063\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e\\u003cp\\u003e0.630\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e\\u003cp\\u003e0.670\\u003c/p\\u003e\\u003c/td\\u003e\\u003c/tr\\u003e\\u003c/tbody\\u003e\\u003c/colgroup\\u003e\\u003ctfoot\\u003e\\u003ctr\\u003e\\u003ctd colspan=\\\"6\\\"\\u003e\\u003csup\\u003ea,b\\u003c/sup\\u003e Values within a column with different superscripts differ significantly at P\\u0026thinsp;\\u0026lt;\\u0026thinsp;0.05.\\u003c/td\\u003e\\u003c/tr\\u003e\\u003c/tfoot\\u003e\\u003c/table\\u003e\\u003c/div\\u003e\\u003c/p\\u003e\"},{\"header\":\"Discussion\",\"content\":\"\\u003cp\\u003eThe current study found that the highest percentage of hatch was related to the control treatment and the average weight of chicks at the time of hatching was not affected by the treatments. In another study, Fazli et al. (\\u003cspan citationid=\\\"CR10\\\" class=\\\"CitationRef\\\"\\u003e2015\\u003c/span\\u003e) reported that \\u003cem\\u003ein-ovo\\u003c/em\\u003e injection of garlic extract did not seem to affect the hatched chick weight. Furthermore, GEO injected into eggs could not improve body weight in chicks at hatch (Mahjar and Al-Salhie \\u003cspan citationid=\\\"CR16\\\" class=\\\"CitationRef\\\"\\u003e2022\\u003c/span\\u003e). On the other hand, \\u003cem\\u003ein-ovo\\u003c/em\\u003e administration with 150 \\u0026micro;l GEO was found to have a higher hatchability than other treatments and decreased mortality in broiler chickens. Researchers indicated that \\u003cem\\u003ein-ovo\\u003c/em\\u003e medical plant injection could increase total antioxidant capacity and decrease free radical production, which would lead to improved hatchability (Radwan Nadia et al. \\u003cspan citationid=\\\"CR22\\\" class=\\\"CitationRef\\\"\\u003e2008\\u003c/span\\u003e; Surai et al. \\u003cspan citationid=\\\"CR29\\\" class=\\\"CitationRef\\\"\\u003e2003\\u003c/span\\u003e). It seems that the positive effect of GEO on hatchability may be due to its beneficial role on embryo development through antioxidant and antibacterial properties (Mahjar and Al-Salhie \\u003cspan citationid=\\\"CR16\\\" class=\\\"CitationRef\\\"\\u003e2022\\u003c/span\\u003e).\\u003c/p\\u003e\\u003cp\\u003eIn the present study, BWG, FI and FCR at day 7 and 14 were not affected by any injection treatments. On the impact of \\u003cem\\u003ein-ovo\\u003c/em\\u003e injection of GEO on chicken performance, however, no research has been found. According to Kumar et al. (\\u003cspan citationid=\\\"CR14\\\" class=\\\"CitationRef\\\"\\u003e2022\\u003c/span\\u003e), adding nanoencapsulated eugenol to garlic extract in broiler diets had no appreciable impact on FI and FCR of the starter/brooding period (0 to 10 days of age) (Kumar et al. \\u003cspan citationid=\\\"CR14\\\" class=\\\"CitationRef\\\"\\u003e2022\\u003c/span\\u003e). Additionally, feeding laying hens garlic had no noticeable impact on their FI (Adebiyi et al., \\u003cspan citationid=\\\"CR2\\\" class=\\\"CitationRef\\\"\\u003e2022\\u003c/span\\u003e). Other studies investigated the effect of garlic supplementation on broiler performance. The results section states that FI, FCR, and BWG were not significantly influenced by experimental treatment (Onibi et al. \\u003cspan citationid=\\\"CR19\\\" class=\\\"CitationRef\\\"\\u003e2009\\u003c/span\\u003e). The reasons may be related to the gastrointestinal tract and the gut enzyme secretion of broiler chickens which are not fully developed during this period (0 to 14 days of age).\\u003c/p\\u003e\\u003cp\\u003eThe result of the present study shows that there were no significant differences in glycogen levels in both tissues and breast muscle among groups of zero-day chickens. The amount of breast glycogen was improved by the injection of nanoencapsulated GEO, but the amount of liver glycogen was not affected by the treatments. Muscle glycogen levels in poultry meat are closely associated with pH and meat technological quality (Scheffler et al. \\u003cspan citationid=\\\"CR27\\\" class=\\\"CitationRef\\\"\\u003e2013\\u003c/span\\u003e). It is proven that low levels of muscle glycogen indicate a decrease in meat pH (Sugiharto \\u003cspan citationid=\\\"CR28\\\" class=\\\"CitationRef\\\"\\u003e2022\\u003c/span\\u003e).Pre-slaughter stress reduces glycogen stores in poultry muscles and could affect the physical and chemical properties of broiler meat (Sugiharto \\u003cspan citationid=\\\"CR28\\\" class=\\\"CitationRef\\\"\\u003e2022\\u003c/span\\u003e). Also, in a study, Kumar et al. (\\u003cspan citationid=\\\"CR15\\\" class=\\\"CitationRef\\\"\\u003e2019\\u003c/span\\u003e) reported that garlic supplementation increases the glycogen of broiler muscle. On the other hand, garlic has a variety of bioactive compounds like flavonoids that may increase the glycogen content of the liver with antioxidant properties (Kumar et al. \\u003cspan citationid=\\\"CR15\\\" class=\\\"CitationRef\\\"\\u003e2019\\u003c/span\\u003e).\\u003c/p\\u003e\\u003cp\\u003eIn this trial, nanoencapsulated GEO significantly improved intestinal morphology and microbial population. The villus height and villus height to crypt depth ratios were considerably higher in the 0.1 ml of nanoencapsulated GEO group compared to control group. Also, the \\u003cem\\u003ein-ovo\\u003c/em\\u003e injection of nanoencapsulated GEO decreased \\u003cem\\u003eE. coli\\u003c/em\\u003e counts compared to both control groups. Consistent with the results of the present study, Karangiya et al. (\\u003cspan citationid=\\\"CR13\\\" class=\\\"CitationRef\\\"\\u003e2016\\u003c/span\\u003e) found that the use of 1% garlic in broiler feed significantly increased the villus height. In another study, researchers found that the height, width, and area of the jejunum and duodenum villus increased in broiler chickens supplemented with garlic compared to control (Bahadoran et al. \\u003cspan citationid=\\\"CR6\\\" class=\\\"CitationRef\\\"\\u003e2016\\u003c/span\\u003e). According to Amiri et al. (\\u003cspan citationid=\\\"CR4\\\" class=\\\"CitationRef\\\"\\u003e2021\\u003c/span\\u003e), a dietary of 100 mg/kg nanoencapsulated GEO showed a higher amount of villus height and villus height to crypt depth ratios in broiler intestine.\\u003c/p\\u003e\\u003cp\\u003eInterestingly, garlic exhibits higher activity against gram-negative and gram-positive bacteria (Munir \\u003cspan citationid=\\\"CR18\\\" class=\\\"CitationRef\\\"\\u003e2015\\u003c/span\\u003e). In vitro and in vivo, GEO nanoencapsulation with chitosan had significantly higher antibacterial activity against \\u003cem\\u003eE. coli\\u003c/em\\u003e compared to free form (Amiri et al. \\u003cspan citationid=\\\"CR4\\\" class=\\\"CitationRef\\\"\\u003e2021\\u003c/span\\u003e). It may be suggested that nanoencapsulated GEO has antioxidant properties (Jakubcov\\u0026aacute; et al. \\u003cspan citationid=\\\"CR12\\\" class=\\\"CitationRef\\\"\\u003e2014\\u003c/span\\u003e) in addition to antimicrobial activity (D\\u0026aacute;vila-Rodrguez et al. 2020). Moreover, it is noteworthy that nanoencapsulation is an improvement method that allows precise and controlled release of bioactive compounds during digestion, which could protect the antibacterial and antioxidant properties of essential oils (Amiri et al. \\u003cspan citationid=\\\"CR4\\\" class=\\\"CitationRef\\\"\\u003e2021\\u003c/span\\u003e).\\u003c/p\\u003e\\u003cp\\u003eAt hatch day, serum glucose increased in the free GEO and control treatments. Although a similar study was not found on the effect of \\u003cem\\u003ein-ovo\\u003c/em\\u003e injection of GEO on broiler blood parameters, the researchers noticed that adding garlic extract to broiler diets had no significant effect on blood cholesterol levels (Brz\\u0026oacute;ska et al. \\u003cspan citationid=\\\"CR7\\\" class=\\\"CitationRef\\\"\\u003e2015\\u003c/span\\u003e). Furthermore, Parvin et al. (\\u003cspan citationid=\\\"CR21\\\" class=\\\"CitationRef\\\"\\u003e2021\\u003c/span\\u003e) stated that garlic inclusion in broiler diets significantly reduces blood glucose. In addition, a report indicated that garlic powder in the broiler diet did not significantly change blood cholesterol concentration compared with the control group (Rahimi et al. \\u003cspan citationid=\\\"CR23\\\" class=\\\"CitationRef\\\"\\u003e2021\\u003c/span\\u003e). Glucose levels may be affected by organosulfur compounds like allicin from garlic (Onunkwo et al. \\u003cspan citationid=\\\"CR20\\\" class=\\\"CitationRef\\\"\\u003e2019\\u003c/span\\u003e). Allicin's mode of action is through competing with insulin for its insulin-inactivating sites in the liver, resulting in lower glucose levels (Ala Al Deen 2007). On the other hand, it has been suggested that the compounds in garlic may induce antagonistic activity with insulin and lead to a slight increase in glucose levels (Ala Al Deen 2007).\\u003c/p\\u003e\"},{\"header\":\"Conclusion\",\"content\":\"\\u003cp\\u003eAccording to the results, \\u003cem\\u003ein-ovo\\u003c/em\\u003e injection of GEO into the yolk sac at day 7 of incubation period reduced the hatching rate. The nanoencapsulated GEO increased breast muscle glycogen, improved intestinal villi, as well as reduced pathogenic populations like \\u003cem\\u003eE. coli.\\u003c/em\\u003e In general, nanocapsulation with chitosan increased the effectiveness of GEO.\\u003c/p\\u003e\"},{\"header\":\"Abbreviations\",\"content\":\"\\u003cp\\u003eGEO, garlic essential oil; EO, essential oils; TPP, sodium triphosphate pentabasic; FI, feed intake; BWG, body weight gain; FCR, feed conversion ratio.\\u003c/p\\u003e\"},{\"header\":\"Declarations\",\"content\":\"\\u003cp\\u003e\\u003ch2\\u003eCompliance with Ethical Standards\\u003c/h2\\u003e\\u003cp\\u003eFunding\\u003c/p\\u003e\\u003c/p\\u003e\\u003cp\\u003e\\u003ch2\\u003eConflict of Interest\\u003c/h2\\u003e\\u003cp\\u003eThe authors declare that they have no conflict of interest.\\u003c/p\\u003e\\u003c/p\\u003e\\u003cp\\u003e\\u003cstrong\\u003eEthical approval\\u003c/strong\\u003e\\u003cp\\u003eThe experiment was conducted according to the animal welfare guidelines at the Veterinary Control and Research Institute of Kerman, Iran (approval number: IR.UK.VETMED.REC 2019-03-05).\\u003c/p\\u003e\\u003c/p\\u003e\\u003cp\\u003e\\u003cstrong\\u003eConsent for publication\\u003c/strong\\u003e\\u003cp\\u003eFor this type of study, consent for publication is not required.\\u003c/p\\u003e\\u003c/p\\u003e\\u003ch2\\u003eAuthor Contribution\\u003c/h2\\u003e\\u003cp\\u003eM.A. conceived and designed the experiment, supervised the project, contributed to data interpretation, and wrote the main manuscript text. N.A. conducted the animal trial, collected samples, and performed the initial data analysis. M.K.B. contributed to statistical analysis and assisted in interpreting the results and manuscript writing. All authors reviewed and approved the final version of the manuscript.\\u003c/p\\u003e\\u003ch2\\u003eData availability statement\\u003c/h2\\u003e\\u003cp\\u003eData will be made available on request.\\u003c/p\\u003e\"},{\"header\":\"References\",\"content\":\"\\u003col\\u003e\\n\\u003cli\\u003eAbd El-Latif AS, Saleh NS, Allam TS, Ghazy EW (2013) The effects of rosemary (Rosemarinus afficinalis) and garlic (Allium sativum) essential oils on performance, hematological, biochemical and immunological parameters of broiler chickens. Br J Poult Sci 2 (2): 16-24. https://doi.org/10.5829/idosi.bjps.2013.2.2.74145.\\u003c/li\\u003e\\n\\u003cli\\u003eAdebiyi FG, Adebiyi OA, Adeyemo GO (2022) Performance and haematological indices evaluation of laying hens fed graded levels of raw garlic. Niger J Anim Prod 49 (2):157-167, https://doi.org/10.51791/njap.v49i2.3475. \\u003c/li\\u003e\\n\\u003cli\\u003eAla Al Deen HJ (2007) Some haematological and biochemical effects of garlic on broiler chicken. Bas J Vet Res 6 (2). https://doi.org/10.33762/bvetr.2007.55779.\\u003c/li\\u003e\\n\\u003cli\\u003eAmiri N, Afsharmanesh M, Salarmoini M, Meimandipour A, Hosseini SA, Ebrahimnejad H (2021) Nanoencapsulation (in vitro and in vivo) as an efficient technology to boost the potential of garlic essential oil as alternatives for antibiotics in broiler nutrition. Animal 15\\u003cem\\u003e \\u003c/em\\u003e(1):100022. https://doi.org/10.1016/j.animal.2020.100022.\\u003c/li\\u003e\\n\\u003cli\\u003eAviagen (2014). Nutrition specifications: ROSS 308 Broiler. Aviagen Ltd.\\u003c/li\\u003e\\n\\u003cli\\u003eBahadoran S, Hasanpour H, Shafigh Z (2016) Garlic supplement improves intestinal mucosa morphology in broiler chickens with developmental pulmonary hypertension. Poult Sci J 4\\u003cem\\u003e \\u003c/em\\u003e(2):117-125. \\u003c/li\\u003e\\n\\u003cli\\u003eBrz\\u0026oacute;ska F, Sliwinski B, Michalik-Rutkowska O, Sliwa J (2015) The effect of garlic (\\u003cem\\u003eAllium sativum\\u003c/em\\u003e L.) on growth performance, mortality rate, meat and blood parameters in broilers. Ann Anim Sci 15 (4):961-975. https://doi.org/10.1515/aoas-2015-0052.\\u003c/li\\u003e\\n\\u003cli\\u003eD\\u0026aacute;vila-Rodr\\u0026iacute;guez M, L\\u0026oacute;pez-Malo A, Palou E, Ram\\u0026iacute;rez-Corona N, Jim\\u0026eacute;nez-Mungu\\u0026iacute;a MT (2020) Essential oils microemulsions prepared with high-frequency ultrasound: Physical properties and antimicrobial activity. J Food Sci Technol 57 (11):4133-4142. https://doi.org/10.1007/s13197-020-04449-8. \\u003c/li\\u003e\\n\\u003cli\\u003eDehghani N, Afsharmanesh M, Salarmoini M, Ebrahimnejad H, Bitaraf (2018) A Effect of pennyroyal, savory and thyme essential oils on Japanese quail physiology. Heliyon 4, e00881. https://doi.org/10.1016/j.heliyon.2018.e00881 \\u003c/li\\u003e\\n\\u003cli\\u003eFazli N, Hassanabadi A, Mottaghitalab M, Hajati H (2015) Manipulation of broiler chickens sex differentiation by in ovo injection of aromatase inhibitors, and garlic and tomato extracts. Poult Sci 94 (11):2778-2783. https://doi.org/10.3382/ps/pev236. \\u003c/li\\u003e\\n\\u003cli\\u003eJafari SZ, Jafarian S, Hojjati M, Najafian L (2022) Evaluation of antioxidant activity of nano‐and microencapsulated rosemary (Rosmarinus officinalis L.) leaves extract in cress (Lepidium sativum) and basil (Ocimum basilicum) seed gums for enhancing oxidative stability of sunflower oil. Food Sci Nutr 10 (6):2111-2119. https://doi.org/10.1002/fsn3.2827. \\u003c/li\\u003e\\n\\u003cli\\u003eJakubcov\\u0026aacute; Z, Mare\\u0026scaron; P, Zeman L, Hork\\u0026yacute; P, Jur\\u0026iacute;kov\\u0026aacute; T, Mlček J, Balla \\u0026Scaron;, Kalhotka L, Mrkvicov\\u0026aacute; E, Sochor J (2014) Influence of garlic extract on antioxidant status of chicken. Potravinarstvo 8 (1):315-320. https://doi.org/10.5219/404. \\u003c/li\\u003e\\n\\u003cli\\u003eKarangiya VK, Savsani HH, Patil SS, Garg DD, Murthy KS, Ribadiya NK, Vekariya SJ (2016) Effect of dietary supplementation of garlic, ginger and their combination on feed intake, growth performance and economics in commercial broilers. Vet World 9 (3):245. https://doi.org/10.14202/vetworld.2016.245-250. \\u003c/li\\u003e\\n\\u003cli\\u003eKumar A, Sharma NK, Kheravii SK, Keerqin C, Ionescu C, Blanchard A, Wu SB (2022) Potential of a mixture of eugenol and garlic tincture to improve performance and intestinal health in broilers under necrotic enteritis challenge. Anim Nutr 8 (1):26-37. https://doi.org/10.1016/j.aninu.2021.07.007. \\u003c/li\\u003e\\n\\u003cli\\u003eKumar R, Maan NS, Baloda S, Dahiya R, Sihag S (2019) Influence on the performance of broilers with the Garlic (Allium sativum) and Holy basil (Ocimum sanctum) leaf powder supplementation in the basal diet. Pharma Innovation 8 (1):553-557. \\u003c/li\\u003e\\n\\u003cli\\u003eMahjar NT, Al-Salhie KC (2022) The effects of in ovo injection of garlic (\\u003cem\\u003eallium sativum l\\u003c/em\\u003e.) extract on hatchability, liver enzymes and antioxidant status of broiler chickens. 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Niger J Anim Prod 46 (1):149-156, https://doi.org/10.51791/njap.v46i1.1312. \\u003c/li\\u003e\\n\\u003cli\\u003eParvin MM, Beg MA, Begum M, Islam MS, Rubel MZ (2021) Effects of using garlic (Allium sativum) and turmeric (Curcuma longa) powder on production performance and biochemical parameters of broiler chicken. Int J Vet Sci Anim Hus 6 (4):53-57. https://doi.org/10.22271/veterinary.2021.v6.i4a.369. \\u003c/li\\u003e\\n\\u003cli\\u003eRadwan Nadia L, Hassan RA, Qota EM, Fayek HM (2008) Effect of natural antioxidant on oxidative stability of eggs and productive and reproductive performance of laying hens. Int J Poult Sci 7 (2):134-150. https://doi.org/10.3923/ijps.2008.134.150. \\u003c/li\\u003e\\n\\u003cli\\u003eRahimi D, Yarahmadi HM, Yaghobfar A, Fakhraei J (2021) Effects of garlic powder and satureja khuzestanica essential oil on male ross 308 chickens performance, blood lipid profile, immune responses, intestinal microflora, and morphology. Jundishapur J Nat Pharm Prod 16 (1). https://doi.org/10.5812/jjnpp.94567. \\u003c/li\\u003e\\n\\u003cli\\u003eRichmond W (1973) Preparation and properties of a cholesterol oxidase from Nocardia sp. and its application to the enzymatic assay of total cholesterol in serum. Clin Chem 19 (12):1350-1356. https://doi.org/10.1093/clinchem/19.12.1350.\\u003c/li\\u003e\\n\\u003cli\\u003eSatyal P, Craft JD, Dosoky NS, Setzer WN (2017) The chemical compositions of the volatile oils of garlic (Allium sativum) and wild garlic (Allium vineale). Foods 6 (8):63. https://doi.org/10.3390/foods6080063.\\u003c/li\\u003e\\n\\u003cli\\u003eSawtarie N, Cai Y, Lapitsky Y (2017) Preparation of chitosan/tripolyphosphate nanoparticles with highly tunable size and low polydispersity. Colloids Surf B 157:110-117. https://doi.org/10.1016/j.colsurfb.2017.05.055. \\u003c/li\\u003e\\n\\u003cli\\u003eScheffler TL, Scheffler JM, Kasten SC, Sosnicki AA, Gerrard DE (2013) High glycolytic potential does not predict low ultimate pH in pork. Meat Sci 95 (1):85-91. https://doi.org/10.1016/j.meatsci.2013.04.013. \\u003c/li\\u003e\\n\\u003cli\\u003eSugiharto S (2022) Potential Benefits of Plant-Derived Products on Broiler Meat Characteristics-A Short Review. J. sain peternak. Indones. 17 (1):29-43. https://doi.org/10.1016/j.meatsci.2013.04.013. \\u003c/li\\u003e\\n\\u003cli\\u003eSurai AP, Surai PF, Steinberg W, Wakeman WG, Speake BK, Sparks NH (2003) Effect of canthaxanthin content of the maternal diet on the antioxidant system of the developing chick. Br Poult Sci 44 (4):612-619. https://doi.org/10.1080/00071660310001616200.\\u003c/li\\u003e\\n\\u003cli\\u003eTako E, Ferket PR, Uni Z (2004) Effects of in ovo feeding of carbohydrates and beta-hydroxy-beta-methylbutyrate on the development of chicken intestine. Poult Sci 83\\u003cem\\u003e \\u003c/em\\u003e(12):2023-2028. https://doi.org/10.1093/ps/83.12.2023. \\u003c/li\\u003e\\n\\u003cli\\u003eYang CM, Cao GT, Ferket PR, Liu TT, Zhou L, Zhang L, Xiao YP, Chen AG (2012) Effects of probiotic, Clostridium butyricum, on growth performance, immune function, and cecal microflora in broiler chickens. Poult Sci 91:2121-2129. https://doi.org/10.3382/ps.2011-02131. \\u003c/li\\u003e\\n\\u003cli\\u003eYazdanpanah Goharrizi L, Tasharofi S (2017) Inhibiting Aspergillus flavus growth and aflatoxin decrement in aquatic feed with Satureja hortensis essen-tial oil. Res J Vet Pract 4 (4):71-75. https://doi.org/10.14737/journal.rjvp/2016/4.4.71.75. \\u003c/li\\u003e\\n\\u003c/ol\\u003e\"}],\"fulltextSource\":\"\",\"fullText\":\"\",\"funders\":[],\"hasAdminPriorityOnWorkflow\":false,\"hasManuscriptDocX\":true,\"hasOptedInToPreprint\":true,\"hasPassedJournalQc\":\"\",\"hasAnyPriority\":false,\"hideJournal\":false,\"highlight\":\"\",\"institution\":\"\",\"isAcceptedByJournal\":true,\"isAuthorSuppliedPdf\":false,\"isDeskRejected\":\"\",\"isHiddenFromSearch\":false,\"isInQc\":false,\"isInWorkflow\":false,\"isPdf\":false,\"isPdfUpToDate\":true,\"isWithdrawnOrRetracted\":false,\"journal\":{\"display\":true,\"email\":\"info@researchsquare.com\",\"identity\":\"comparative-clinical-pathology\",\"isNatureJournal\":false,\"hasQc\":true,\"allowDirectSubmit\":false,\"externalIdentity\":\"\",\"sideBox\":\"Learn more about [Comparative Clinical Pathology](https://link.springer.com/journal/580)\",\"snPcode\":\"580\",\"submissionUrl\":\"https://submission.springernature.com/new-submission/580/3\",\"title\":\"Comparative Clinical Pathology\",\"twitterHandle\":\"\",\"acdcEnabled\":true,\"dfaEnabled\":true,\"editorialSystem\":\"stoa\",\"reportingPortfolio\":\"Springer Hybrid\",\"inReviewEnabled\":true,\"inReviewRevisionsEnabled\":false},\"keywords\":\"In-ovo injection, Yolk sac, Garlic essential oil, Nano-encapsulation, Broiler chicken\",\"lastPublishedDoi\":\"10.21203/rs.3.rs-7119473/v1\",\"lastPublishedDoiUrl\":\"https://doi.org/10.21203/rs.3.rs-7119473/v1\",\"license\":{\"name\":\"CC BY 4.0\",\"url\":\"https://creativecommons.org/licenses/by/4.0/\"},\"manuscriptAbstract\":\"\\u003cp\\u003eThe purpose of this research was to determine the impact of \\u003cem\\u003ein-ovo\\u003c/em\\u003e injection of garlic essential oil (GEO) on post-hatch broiler chicken performance, intestinal development, hatch percentage, and blood parameters. 360 Ross 308 eggs weighing 62\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;2 g were selected and 15 eggs per replicate were used in a completely randomized design. The experimental treatments were as follows: (1) control or non-injected, (2) \\u003cem\\u003ein-ovo\\u003c/em\\u003e injection of physiological saline or positive control, (3) \\u003cem\\u003ein-ovo\\u003c/em\\u003e injection of 0.1 free GEO, (4) 0.2 free GEO, (5) 0.1 nanoencapsulated GEO, (6) 0.2 nanoencapsulated GEO into the yolk sac at day 7 of incubation period. In contrast to the control treatment (injection-free), the results demonstrated that \\u003cem\\u003ein-ovo\\u003c/em\\u003e injection of free GEO lowered hatch percentage (P\\u0026thinsp;\\u0026lt;\\u0026thinsp;0.05). The post-hatch broiler chicken performance was unaffected by different treatments. The breast glycogen concentration and the villus height and villus height to crypt depth ratios were higher in the 0.1 ml of nanoencapsulated GEO group (P\\u0026thinsp;\\u0026lt;\\u0026thinsp;0.05). The \\u003cem\\u003ein-ovo\\u003c/em\\u003e injection of nanoencapsulated GEO decreased \\u003cem\\u003eE. coli\\u003c/em\\u003e counts compared to control group (P\\u0026thinsp;\\u0026lt;\\u0026thinsp;0.05). In conclusion, chitosan nanoencapsulation has the potential to protect bioactive substances and enhance the benefits of GEO in newly hatched chickens.\\u003c/p\\u003e\",\"manuscriptTitle\":\"Impact of in ovo feeding of nano-encapsulated garlic essential oil on broiler hatch percentage, performance, intestinal development, and blood parameters\",\"msid\":\"\",\"msnumber\":\"\",\"nonDraftVersions\":[{\"code\":1,\"date\":\"2025-08-28 05:59:17\",\"doi\":\"10.21203/rs.3.rs-7119473/v1\",\"editorialEvents\":[{\"type\":\"communityComments\",\"content\":0},{\"type\":\"decision\",\"content\":\"Revision requested\",\"date\":\"2025-09-07T16:52:34+00:00\",\"index\":\"\",\"fulltext\":\"\"},{\"type\":\"editorInvitedReview\",\"content\":\"\",\"date\":\"2025-09-06T17:41:08+00:00\",\"index\":\"hide\",\"fulltext\":\"\"},{\"type\":\"editorInvitedReview\",\"content\":\"\",\"date\":\"2025-08-30T10:11:37+00:00\",\"index\":\"hide\",\"fulltext\":\"\"},{\"type\":\"reviewerAgreed\",\"content\":\"209588686073637144079046845482543406536\",\"date\":\"2025-08-26T10:48:28+00:00\",\"index\":\"hide\",\"fulltext\":\"\"},{\"type\":\"reviewerAgreed\",\"content\":\"244731638653897041687620756016499721496\",\"date\":\"2025-08-20T10:07:43+00:00\",\"index\":\"hide\",\"fulltext\":\"\"},{\"type\":\"reviewersInvited\",\"content\":\"\",\"date\":\"2025-08-20T09:46:24+00:00\",\"index\":\"\",\"fulltext\":\"\"},{\"type\":\"editorAssigned\",\"content\":\"\",\"date\":\"2025-07-16T15:09:51+00:00\",\"index\":\"\",\"fulltext\":\"\"},{\"type\":\"checksComplete\",\"content\":\"\",\"date\":\"2025-07-16T15:08:14+00:00\",\"index\":\"\",\"fulltext\":\"\"},{\"type\":\"submitted\",\"content\":\"Comparative Clinical Pathology\",\"date\":\"2025-07-14T09:24:08+00:00\",\"index\":\"\",\"fulltext\":\"\"}],\"status\":\"published\",\"journal\":{\"display\":true,\"email\":\"info@researchsquare.com\",\"identity\":\"comparative-clinical-pathology\",\"isNatureJournal\":false,\"hasQc\":true,\"allowDirectSubmit\":false,\"externalIdentity\":\"\",\"sideBox\":\"Learn more about [Comparative Clinical Pathology](https://link.springer.com/journal/580)\",\"snPcode\":\"580\",\"submissionUrl\":\"https://submission.springernature.com/new-submission/580/3\",\"title\":\"Comparative Clinical Pathology\",\"twitterHandle\":\"\",\"acdcEnabled\":true,\"dfaEnabled\":true,\"editorialSystem\":\"stoa\",\"reportingPortfolio\":\"Springer Hybrid\",\"inReviewEnabled\":true,\"inReviewRevisionsEnabled\":false}}],\"origin\":\"\",\"ownerIdentity\":\"c692b4fa-b4ef-44d6-b58d-1230f5b3b509\",\"owner\":[],\"postedDate\":\"August 28th, 2025\",\"published\":true,\"recentEditorialEvents\":[],\"rejectedJournal\":[],\"revision\":\"\",\"amendment\":\"\",\"status\":\"under-review\",\"subjectAreas\":[],\"tags\":[],\"updatedAt\":\"2025-12-07T07:38:29+00:00\",\"versionOfRecord\":[],\"versionCreatedAt\":\"2025-08-28 05:59:17\",\"video\":\"\",\"vorDoi\":\"\",\"vorDoiUrl\":\"\",\"workflowStages\":[]},\"version\":\"v1\",\"identity\":\"rs-7119473\",\"journalConfig\":\"researchsquare\"},\"__N_SSP\":true},\"page\":\"/article/[identity]/[[...version]]\",\"query\":{\"redirect\":\"/article/rs-7119473\",\"identity\":\"rs-7119473\",\"version\":[\"v1\"]},\"buildId\":\"8U1c8b4HqxoKbykW_rLl7\",\"isFallback\":false,\"isExperimentalCompile\":false,\"dynamicIds\":[84888],\"gssp\":true,\"scriptLoader\":[]}","source_license":"CC-BY-4.0","license_restricted":false}