Research Article

Interactive Effects of Egg Mass and Pre-hatch Injection of Alcoholic Date Palm Pollen Extract on Hatch Traits of Broiler Chickens (Ross 308)

Ban Salman Kadhim*, Basil Mohammed Ibrahim

Department of Animal Production, College of Agricultural Engineering Sciences, University of Baghdad, Baghdad, Iraq.

Received | March 03, 2026; Accepted | April 15, 2026; Published | July 07, 2026

*Correspondence | Ban Salman Kadhim, Department of Animal Production, College of Agricultural Engineering Sciences, University of Baghdad, Baghdad, Iraq; Email: [email protected]

Citation | Kadhim BS, Ibrahim BM (2026). Interactive effects of egg mass and pre-hatch injection of alcoholic date palm pollen extract on hatch traits of broiler chickens (Ross 308). J. Anim. Health Prod. 14(3): 1056-1062.

DOI | https://dx.doi.org/10.17582/journal.jahp/2026/14.3.1056.1062

ISSN (Online) | 2308-2801

Copyright: 2026 by the authors. Licensee ResearchersLinks Ltd, England, UK.

This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

Introduction

Hatchability and productive traits of broiler chickens are influenced by both genetic and environmental factors. Among these, internal and external egg quality traits are particularly important, and egg weight is a key parameter associated with egg quality due to its effect on hatchability and broiler performance (Ulmer-Franco et al., 2010; Khalil et al., 2022). A strong relationship exists between egg weight and hatchability traits, as it influences hatchability percentage, incubation duration, embryonic mortality rate, and chick weight at hatch (Alabi et al., 2012; Ajafar et al., 2024). Some studies have reported that higher egg weights may reduce fertility and hatchability; however, this effect is often associated with breeder age rather than egg weight itself (Iqbal et al., 2016).

Recent research has focused on improving embryo weight and immunity at hatching through in ovo feeding techniques, which enhance embryonic nutrition and compensate for nutrient losses during incubation. Providing essential nutrients directly to the embryo can improve development and hatch performance, contributing to sustainable poultry production (Du et al., 2025; Arkan et al., 2025). However, a major limitation of this approach is the potential reduction in hatchability due to the injection process, which is generally undesirable in commercial incubation. Therefore, it is important to optimize in ovo supplementation strategies that do not negatively affect hatchability. Several studies have evaluated the effects of injecting nutritional compounds into eggs, but results remain inconsistent, with many reporting reduced hatchability (Al-Shammari, 2025).

Various nutritional components have been shown to enhance embryonic growth and weight (Ma et al., 2020; Dos et al., 2010). Nevertheless, the exploration of novel in ovo feeding substances remains essential for improving embryonic development. Date palm pollen, the male reproductive structure of the date palm, has traditionally been used in folk medicine and was once referred to as the “Fountain of Youth” due to its reputed health benefits (Okai et al., 2025).

Several studies have demonstrated that date palm pollen possesses medicinal properties attributable to its high protein content, which supports muscle and tissue development. It also contains antioxidants and healthy fats that help protect against oxidative stress and maintain physiological health (Salhi et al., 2024).

Due to these properties, date palm pollen has been used as a supplement in poultry production. Studies have reported that it can enhance egg production and improve egg quality in laying hens, likely due to its phytoestrogen (estradiol-like) compounds that regulate reproductive functions and improve production efficiency (Mousa et al., 2018). Additionally, date palm pollen has been used in in ovo applications to enhance embryonic development, hatchability, and chick weight at hatch (Coşkun et al., 2014; Salman et al., 2025; Alsudani et al., 2025).

Therefore, this study aimed to investigate the effects of egg weight and in ovo injection of alcoholic extract of date palm pollen on hatchability traits of Ross 308 broiler chickens.

Materials and Methods

Ethical approval

The present research was performed in compliance with the principles of scientific research ethics and within the rules and under the observation and approval of the Research Ethics Committee of the Animal Production Department, College of Agricultural Engineering Sciences, University of Baghdad. The hatching eggs used in this experiment were incubated in accordance with the official management guide of the Ross 308 strain, with a suitable environmental condition inside the incubator, taking into consideration the safety of the embryo and avoiding any harm not justified by a cause. Official approvals were obtained before the beginning of the experiment.

Experimental design

A total of 1,000 hatching eggs from 38-week-old Ross 308 broiler breeder hens were obtained from Al-Baraka Hatchery in Wasit Province. The eggs were divided into two groups based on egg weight. The first group consisted of large eggs with an average weight of 61 ± 2 g, while the second group consisted of small eggs with an average weight of 55 ± 2 g. Each group contained 500 eggs.

Each weight group was further subdivided into five treatment groups, with 100 eggs per treatment, as follows: T1, control (no injection); T2, positive control (injection with normal saline); T3, injection with alcoholic extract of date palm pollen (125 µg/mL); T4, injection with alcoholic extract of date palm pollen (150 µg/mL); and T5, injection with alcoholic extract of date palm pollen (175 µg/mL).

Preparation of the extract and injection procedure

Palm pollen grains were obtained from a date palm orchard located in Al-Nu’maniyah District, Wasit Province. The pollen was separated from its protective sheath by sieving, and the fine pollen powder was collected. The alcoholic extract of the palm pollen was prepared at the laboratories of the Ministry of Science and Technology/ Directorate of Environment and Water.

The extraction process was carried out following the method described by Basuny et al. (2013), using 100 g of palm pollen mixed with 500 mL of 80% ethanol in a 1000 mL glass flask at room temperature. The mixture was stirred thoroughly using a magnetic stirrer for 24 hours. Afterward, the solution was filtered using filter paper and a Buchner funnel connected to a vacuum pump. The filtrate was then transferred to a centrifuge operating at 3000 rpm for 10 minutes to obtain a clear solution.

 

Table 1: The active compounds present in date palm pollen extract (ppm).

Active compounds (ppm)	B-sterol	Stigma-sterol	Gallic acid	Ferulic acid	Rutin	Apigenin	Syringeic acid	Qurcetine	caffeic acid
date palm pollen extract	9.08	17.90	98.0	60.3	87.4	64.9	70.8	80.2	74.9

 

The resulting supernatant was dried using a rotary evaporator, yielding sufficient quantities of dry extract, which were stored in dark-colored containers at refrigeration temperature until used for preparing injection dilutions.

Sterol and phenolic compounds present in the prepared extract were quantified (Table 1). β-sitosterol and stigmasterol were estimated according to the method of (Maji et al., 2014), while phenolic compounds were determined following the procedure of (Mradu et al., 20١٢). Identification of compounds was based on comparing the chromatographic profiles of standard solutions obtained by HPLC with those of the extract, using retention time alignment.

The dilutions were prepared at the laboratories of the College of Agricultural Engineering Sciences, University of Baghdad. The required quantities of the extract for each treatment (25, 30, and 35 mg) were weighed using a precision electronic balance. Each quantity was then mixed with 200 mL of normal saline to prepare the target concentrations for the experimental treatments (125, 150, and 175 µg/ml, respectively). The solutions were homogenized using a magnetic stirrer. The prepared solutions were then stored in brown glass bottles under refrigeration until use. The treated eggs were injected of the prepared solution 0.2 ml per egg (Al-Hatheel, 2022) once only into the air cell prior to incubation.

Studied hatching traits

• The hatching percentage of the total eggs (%) = (Number of hatched chicks/ Total number of eggs) × 100 (Hussain et al., 2020).
• Embryonic mortality percentage (%) = (Number of embryonic mortalities/ Number of fertile eggs) × 100 (Refaie et al., 2022).
• Egg weight loss percentage (%) = [(Egg Weight before incubation – Egg Weight at day 18)/ Egg Weight before incubation] × 100 (Tona et al., 2022).

Statistical analysis

The data were analyzed using a Completely Randomized Design (CRD) to study the effect of two factors on the experimental units across various traits, employing the Statistical Analysis System (SAS, 2013). Significant differences between means were determined using Duncan’s multiple range test (Duncan, 1955).

Results

Chick-to-egg weight and egg weight loss percentage

As showed in Table 2, incubation period (1-18 days) was significantly influenced by egg weight as well as treatment. Highly significant reduction (p < 0.01) was noticed in chick to egg weight percentage in Group 2 over Group 1. At the same time, loss of egg weight during incubation was significantly lower in Group 1 (9.45%) as compared to Group 2 (10.21%) (p < 0.05). Among the treatments, the differences that proved to be significant were those in the egg weight loss percentage (P < 0.05),

 

Table 2: Effect of egg weight and injection treatment with alcoholic extract of date palm pollen, and their interaction, on the chick to egg weight percentage and egg loss percentage during the incubation period (1-18 day) (mean ± SE).

Effect factors		Chick weight/ egg weight (%)	Egg loss (1- 18 day, %)
Groups	1	71.43±0.35a	9.45±0.14b
	2	69.96±0.38b	10.21±0.20a
Sig.		**	*
Treatments	T1	70.97±0.36	9.81±0.27ab
	T2	69.58±0.87	10.22±0.37a
	T3	70.51±0.38	9.37±0.30b
	T4	71.02±0.61	9.59±0.21ab
	T5	70.72±0.73	10.14±0.32ab
Sig.		N.S	*
Group1	T1	70.92±0.51ab	9.53±0.47bc
	T2	70.87±0.76ab	9.77±0.20abc
	T3	71.23±0.37ab	9.17±0.48c
	T4	72.08±0.34a	9.28±0.05c
	T5	72.06±0.63a	9.47±0.23bc
Group2	T1	71.02±0.63ab	10.09±0.26abc
	T2	68.29±1.25c	10.68±0.66ab
	T3	69.79±0.27abc	9.57±0.41bc
	T4	69.97±0.79abc	9.90±0.35abc
	T5	69.38±0.70bc	10.80±0.14a
Sig.		*	*

 

Egg loss percentage during the incubation period from day 1 to day 18. *Different superscript letters within the same column indicate significant differences among means (P < 0.05). **Different superscript letters indicate highly significant differences among means (P < 0.01). N.S. = not significant (P > 0.05).

 

with the highest value recorded for T2 (10.22%) and the lowest for T3 (9.37%). Non-significant differences were found for the weight percentage of chicks to eggs. The interaction between egg weight and injection treatments showed that T2 in Group 2 had a significantly lowered percentage of chick-to-egg weight compared to the control of the same group and all treatments in Group 1. On the other hand, T5 in Group 2 recorded a significant increase in egg weight loss when compared with T3 within the same group and all treatments of Group 1 except T2.

 

Table 3: Effect of egg weight and injection treatment with alcoholic extract of date palm pollen, and their interaction, on embryonic mortality percentage during three incubation stages (mean ± SE).

Effect factors		M1 (%)	M2 (%)	M3 (%)	TM (%)
Groups	1	1.53±0.51	1.53±0.51a	2.88±0.69	7.46±1.08
	2	0.86±0.38	0.17±0.17b	3.42±0.75	5.82±0.97
Sig.		N.S	*	N.S	N.S
Treatments	T1	0.00±0.00b	0.42±0.42b	2.50±1.01b	5.00±1.41b
	T2	2.23±0.99a	0.00±0.00b	3.13±1.17ab	6.25±1.62ab
	T3	1.25±0.72ab	1.25±0.72ab	5.42±1.46a	10.00±1.94a
	T4	1.25±0.72ab	0.42±0.42b	1.25±0.72b	4.17±1.29b
	T5	1.30±0.75ab	2.17±0.96a	3.48±1.21ab	7.83±1.77ab
Sig.		*	*	*	*
Group1	T1	0.00±0.00b	0.83±0.83b	1.67±1.17b	5.00±2.00b
	T2	3.33±1.65a	0.00±0.00b	3.33±1.65ab	7.50±2.41ab
	T3	0.83±0.83ab	2.50±1.43ab	6.67±2.29a	12.50±3.03a
	T4	2.50±1.43ab	0.00±0.00b	0.83±0.83b	4.17±1.83b
	T5	0.91±0.91ab	4.55±2.00a	1.82±1.82ab	8.18±2.63ab
Group2	T1	0.00±0.00b	0.00±0.00b	3.33±1.65ab	5.00±2.00b
	T2	0.96±0.96ab	0.00±0.00b	2.88±1.65b	4.81±2.11b
	T3	1.67±1.17ab	0.00±0.00b	4.17±1.83ab	7.50±2.41ab
	T4	0.00±0.00b	0.83±0.83b	1.67±1.17ab	4.17±1.83b
	T5	1.67±1.17ab	0.00±0.00b	5.00±2.00ab	7.50±2.41ab
Sig.		*	*	*	*

 

Embryonic mortalities

Embryonic mortality was determined at three levels of incubation: Early incubation (M1: 1-7 days), mid-incubation (M2: 7-14 days), and late incubation (M3: 14-21 days). Analysis of the results presented in Table 3 indicates that egg weight did not significantly influence embryonic mortality in the early (M1) and late (M3) incubation periods; total embryonic mortalities in the two weight categories were not significantly different. But in the mid-incubation stage (M2), there was a significant (P < 0.05) reduction in embryonic mortality in Group 2 relative to that in Group 1. Concerning the impact of treatment, there were significant differences (P < 0.05) among the variables regarding total embryonic mortality (TM). A higher rate was recorded under treatment T3 (10.00 %), and this was significantly different from treatments T1 and T4, with values of 5.00 % and 4.17 %, respectively. Interaction analysis showed that T3 in Group 1 recorded the significantly higher rate of mortality for stages compared with T1 and T4 within the same group, and also compared with T1, T2, and T4 in Group 2. There were no significant differences among other treatment levels.

Dead and live pipped eggs

As presented in Table 4, no significant differences were observed between the two groups in the percentages of dead pipped eggs (PD) or live pipped eggs (PL) (P > 0.05). This indicates that egg weight did not influence the proportion of embryos that failed to emerge or those that successfully pipped but failed to hatch. Similarly, no significant differences were detected among treatments for either PD or PL, suggesting that in ovo injection with alcoholic extract of date palm pollen did not affect hatchability during the pipping stage. The interaction between egg weight and treatment was also not significant, indicating that neither egg size nor extract concentration had a measurable effect on the proportion of dead or live pipped chicks. Overall, the consistently low PD and PL values across all treatments reflect uniform incubation performance. This may be attributed to environmental and genetic uniformity, which likely minimized late-stage embryonic losses.

 

Table 4: Effect of egg weight and treatment with alcoholic extract of date palm pollen, and their interaction, on the percentages of dead and live pipped eggs (mean ± SE).

Effect factors		PD	PL
Groups	1	1.02±0.41	0.51±0.29
	2	0.86±0.38	0.51±0.30
Sig.		N.S	N.S
Treatments	T1	0.38±0.59	1.25±0.72
	T2	0.45±0.45	0.45±0.45
	T3	1.67±0.83	0.42±0.42
	T4	1.25±0.72	0.00±0.00
	T5	0.43±0.43	0.43±0.43
Sig.		N.S	N.S
Group1	T1	0.83±0.83	1.67±1.17
	T2	0.83±0.83	0.00±0.00
	T3	2.50±1.43	0.00±0.00
	T4	0.83±0.83	0.00±0.00
	T5	0.00±0.00	0.91±0.91
Group2	T1	0.83±0.83	0.83±0.83
	T2	0.00±0.00	0.96±0.96
	T3	0.83±0.83	0.83±0.83
	T4	1.67±1.17	0.00±0.00
	T5	0.83±0.83	0.00±0.00
Sig.		N.S	N.S

 

PD = Percentage of dead pecking chicks, PL = Percentage of live pecking chicks, N.S = no significant differences between means (P˃ 0.05) within one column.

 

Hatchability percentage and chick’s weight and length at hatch

As shown in Table 5, egg weight had no significant effect on hatchability percentage (P > 0.05), as similar hatch rates were observed in heavier eggs of Group 1 (85.59%) and lighter eggs of Group 2 (87.33%). However, a significant difference was observed between groups in chick weight at hatch (P < 0.05), with heavier chicks recorded in Group 1 (43.72 g) compared with Group 2 (38.81 g). Chick length at hatch was not significantly influenced by egg weight.

Regarding treatments, hatchability was significantly higher in T1 (90.00%) compared with T3 (82.50%), while no significant differences were observed among the remaining treatments. Chick weight was also significantly affected by treatment (P < 0.05), with T2 showing a noticeable reduction compared with most other treatments, except T3.

 

Table 5: Effect of egg weight and treatment with alcoholic extract of date palm pollen, and their interaction, on hatching percentage, chick weight, and chick length at hatch (mean ± SE).

Effect factors		Hatchability(٪)	Chick's weight at hatch)gm)	Chick's length at hatch(cm)
Groups	1	85.59± 1.45	٤٣.٧٢±٠.٠٥ a	18.36±0.05
	2	87.33±1.38	38.81±0.10b	18.21±0.05
sig		N.S	*	N.S
Treatments	T1	90.00± 1.94a	41.38±1.07a	18.40±0.05
	T2	84.82 ±2.40ab	40.91±1.13b	18.21±0.07
	T3	82.50± 2.46b	41.22±1.09ab	18.28±0.06
	T4	89.17 ± 2.01ab	41.39±1.10a	18.30±0.09
	T5	85.65 ± 2.23ab	41.43±1.12a	18.25±0.13
Sig.		*	*	N.S
Group1	T1	90.83± 2.65a	43.76±0.05ab	18.43±0.07
	T2	81.67±3.55bc	43.43±0.01b	18.28±0.09
	T3	77.50±3.83c	43.65±0.05ab	18.34±0.05
	T4	90.00±2.75ab	43.84±0.09ab	18.48±0.04
	T5	88.18±3.09ab	43.93±0.12a	18.29±0.22
Group2	T1	89.17±2.85ab	39.00±0.30c	18.37±0.09
	T2	88.46±3.15ab	38.39±0.20d	18.14±0.12
	T3	87.50±3.03ab	38.79±0.08cd	18.21±0.09
	T4	88.33±2.94ab	38.95±0.22c	18.12±0.09
	T5	83.33±3.42abc	38.92±0.12c	18.21±0.17
Sig.		*	*	N.S

 

*Different superscript letters within the same column indicate significant differences among means (P < 0.05). N.S. = not significant (P > 0.05).

 

No significant differences were observed in chick length among treatments (P > 0.05). The interaction between treatment and egg weight showed that T3 in Group 1 had lower hatchability compared with most other treatment combinations, except for T2 in Group 1 and T5 in Group 2. For chick weight, treatments within Group 1 exhibited higher values than those in Group 2, indicating a strong association between egg size and chick weight at hatch. In contrast, chick length was not affected by either factor or their interaction, showing no observable interaction effect on this trait.

Discussion

The current study demonstrated that egg weight was the primary factor influencing chick weight and egg weight loss, whereas the effect of in ovo injection of date palm pollen (DPP) extract on hatchability and embryonic mortality was relatively limited. This finding is consistent with previous reports indicating that inherent egg characteristics play a fundamental role in determining embryonic development and chick quality. Larger eggs contain greater nutrient reserves and a larger surface area, which may enhance gas exchange and support water balance during incubation (Grochowska et al., 2019). In contrast, smaller eggs, characterized by reduced yolk content and relatively higher rates of water loss, produced chicks with lower chick-to-egg weight percentage, in agreement with Yousaf et al. (2024). Differences in egg weight loss between groups may also be associated with variations in eggshell conductance, as suggested by Al-Jebory and Ibrahim (2021), Tona et al. (2022), and Al-Saeedi et al. (2023).

Although DPP extract is rich in bioactive compounds such as flavonoids, phenolics, and phytosterols, which possess antioxidant and hormonal regulatory properties (Hussain et al., 2020), the tested concentrations (125–175 µg/ml) did not significantly improve hatchability. This may be attributed to the sensitivity of avian embryos to the plant-derived compounds. Such exposure may induce osmotic stress, where differences in solute concentration disrupt water balance across embryonic membranes, potentially impairing cellular function. In addition, chemical stress may occur, as excessive bioactive compounds could interfere with metabolic processes (Refaie et al., 2022).

The slight increase in embryonic mortality observed at the concentration (T3) suggests that even moderate doses may influence metabolic homeostasis or gas exchange, consistent with findings by El-Kholy et al. (2021). Embryonic mortality may also be linked to reduced embryonic efficiency, as embryos with limited physiological capacity are less able to utilize nutrients and tolerate incubation stress, particularly during the late stages of development (Druyan and Levi, 2012). However, the minor reduction in hatchability was not associated with percentages of dead or live pipped eggs or deformities in hatched chicks, indicating that DPP extract did not adversely affect late embryogenesis or neuromuscular coordination during hatching. These findings agree with previous studies reported that embryos developing under optimal incubation conditions generally hatch normally despite minor treatment effects (Yalcin et al., 2022; Akosile et al., 2023; Oke et al., 2021), and that normal chick morphology reflects the physiological safety of the tested concentrations.

Conclusions

From the results of this research, the fact that egg weight strongly determines the weight of the chicks as well as the egg weight loss can now be affirmed. Despite the fact that the hatchability percentage of eggs was not influenced by the weights of the eggs, the heaviest eggs produced the heaviest chicks and more favorable chick-to-egg weight percentage. The in ovo injection with alcoholic extract of date palm pollen (DPP) did not bring remarkable improvements regarding hatchability. Nevertheless, differences among treatments and control by mortality percentage imply that embryo sensitivity varies with regards to concentration and egg size. Lack of harm caused to chick length and hatched success excludes negative impacts and reveals that none of the DPP doses used had positive effects. Overall, the study proves that the natural properties of the egg, especially weight and the biochemical content, play a more significant role in embryo development and chickes weight at hatched than the external addition of phytochemicals. The selection of eggs and the incubation conditions continue to play the most important critical factors in the production of quality chicks in the broiler production systems. Future studies are required to optimize in ovo injection techniques, establish optimal concentrations of natural bioactive compounds, and determine their molecular effects on embryonic metabolism to improve hatchability and post-hatch performance.

Acknowledgments

The authors would like to thank Al-Baraka Poultry Company (Wasit Governorate, Iraq) for providing the experimental facilities and technical support necessary for conducting this study. Their assistance in egg handling and incubation management is gratefully acknowledged.

Novelty Statement

This study provides a novel evaluation of the combined effects of egg mass and in ovo injection of alcoholic date palm pollen extract on broiler hatch traits. It reveals that egg weight remains the primary determinant of chick quality, while the tested phytochemical intervention shows no added benefit, offering new insight into the limited efficacy of such applications and guiding future optimization of in ovo strategies.

Author’s contribution

BSK: Conducted the experiment and collected the data. BMI: Performed the statistical analysis and critically reviewed the article.

AI and AI-assisted technologies use declaration

No generative AI or AI-assisted technology was used in the preparation of this manuscript.

Conflict of interest

The authors have declared no conflict of interest.

References

Ajafar M, Al-Jebory HH, Al-Saeedi MK (2024). Effect of in ova injection of lysophospholipid in hatching traits, chick’s quality, and chicks physical traits of broiler (Ross 308). Adv. Anim. Vet. Sci., 12: 1206-1213. https://doi.org/10.17582/journal.aavs/2024/12.7.1206.1213

Akosile OA, Kehinde FO, Oni AI, Oke OE (2023). Potential implication of in ovo feeding of Phytogenics in poultry production. Transl. Anim. Sci.,7: 094. https://doi.org/10.1093/tas/txad094

Alabi OJ, Ng’ambi JW, Norris D, Mabelebele M (2012). Effect of egg weight on hatchability and subsequent performance of Potchefstroom Koekoek chicks. Afr. J. Agric. Res., 7: 718–725. https://scialert.net/abstract/?doi=ajava.2012.718.725. https://doi.org/10.3923/ajava.2012.718.725

Al-Hatheel HD (2022). Effect of injecting hatching eggs of aged broiler breeder with alcoholic extract of dried moringa leaf powder Moringa oleifera on hatching traits, quality of hatched chicks and their productive performance. Thesis. College of Agricultural Engineering Sciences. University of Baghdad.

Al-Jebory HH, Ibrahim MK (2021). Effect of adding bee propolis to diet on productive performance of broiler chickens. Indian J. Ecol., 48: 203-207.

Al-Saeedi MK, Al-Jebory HH, Ajafar M (2023). Effect of in ova injection with nano-copper in productive performance of Japanese quail exposed to pathological and environmental challenges. Ann. Agri-Bio Res., 28: 361-366.

Al-Shammari KI (2025). Influence of in ovo injection of phytochemicals and post-hatch feeding on hatching traits, physiological aspects, productive performance, duodenal morphology, and digestibility of broiler chickens. Poult. Sci. J., 13: 215-232.

Alsudani AA, Al-Jebory HH, Al-Saeedi MK (2025). Effects of in-ovo injection of znic-methionine on some microbial and parameters of immunology of broiler. Punjab Univ. J. Zool., 40: 01-07. https://doi.org/10.17582/journal.pujz/2025/40.1.1.7

Arkan NB, Al-Jebory HH, Al-Khafaji FR (2025). Impact of nano-encapsulated of anti-bacterial peptide on broiler growth performance under oxidative stress condition. Adv. Anim. Vet. Sci., 13: 1169-1179. https://doi.org/10.17582/journal.aavs/2025/13.6.1169.1179

Basuny AM, Arafat SM, Soliman HM (2013). Chemical analysis of olive and palm pollen: Antioxidant and antimicrobial activation properties. Wudpecker J. Food Technol., 1:14-21.

Coşkun İ, Çayan H, Yılmaz Ö, Taşkın A, Tahtabiçen E, Samli H (2014). Effects of in ovo pollen extract injection to fertile broiler eggs on hatchability and subsequent chick weight. Turk. J. Agric. Nat. Sci., 1: 485–489.

Dos ST, Corzo A, Kidd MT, McDaniel CD, Torres Filho RA, Araújo LF (2010). Influence of in ovo inoculation with various nutrients and egg size on broiler performance. J. Appl. Poult. Res., 19: 1-12. https://doi.org/10.3382/japr.2009-00038

Druyan S, Levi E (2012). Reduced O2 concentration during CAM development–Its effect on angiogenesis and gene expression in the broiler embryo CAM. Gene Expr. Patterns, 12: 236-244. https://doi.org/10.1016/j.gep.2012.05.001

Du S, Zhou J, Ao X, Zhu Y (2025). Effects of in ovo feeding of vitamin C on embryonic development, hatching process, and chick rectal temperature of broiler embryos. Front. Vet. Sci., 11: 1505801. https://doi.org/10.3389/fvets.2024.1505801

Duncan DB (1955). Multiple range and multiple F tests. Biometrics, 11: 1–42. https://doi.org/10.2307/3001478

El-Kholy KH, Sarhan DM, El-Said EA (2021). Effect of in-ovo Injection of herbal extracts on post-hatch performance, immunological, and physiological responses of broiler chickens. J. World Poult. Res., 11: 183-192. https://doi.org/10.36380/jwpr.2021.22

Grochowska E, Kinal A, Sobek Z, Siatkowski I, Bednarczyk M (2019). Field study on the factors affecting egg weight loss, early embryonic mortality, hatchability, and chick mortality with the use of classification tree technique. Poult. Sci., 98: 3626-3636. https://doi.org/10.3382/ps/pez180

Hussain MI, Farooq M, Syed QA (2020). Nutritional and biological characteristics of the date palm fruit (Phoenix dactylifera L.): A review. Food Biosci., 34: 100509. https://doi.org/10.1016/j.fbio.2019.100509

Iqbal J, Khan SH, Mukhtar N, Ahmed T, Pasha RA (2016). Effects of egg size (weight) and age on hatching performance and chick quality of broiler breeder. J. Appl. Anim. Res., 44: 54-64. https://doi.org/10.1080/09712119.2014.987294

Khalil I, Al-Saeedi MH, Al-Jebory HH, Abood M (2022). Progress phenotypic traits of hatched chicks and growth indicators of broiler chicks fed embryonically with zinc methionine. Arch. Razi. Inst., 77: 2139-2145.

Ma YB, Zhang FD, Wang J, Wu SG, Qi GH, Zhang HJ (2020). Effect of in ovo feeding of β-hydroxy-β-methylbutyrate on hatchability, muscle growth and performance in prenatal and posthatch broilers. J. Sci. Food Agric., 100: 755-763. https://doi.org/10.1002/jsfa.10080

Maji AK, Pandit S, Banerji P, Banerjee D (2014). A validated RP-HPLC method for simultaneous determination of betulin, lupeol, and stigmasterol in Asteracantha longifolia Nees. Int. J. Pharm. Pharm. Sci., 6: 691-695.

Mousa MA, Abedellaah AB, Osman AS, Samer AE (2018). Sparing effects of natural antioxidant derived from two levels of date palm pollen (Phoenix dactylifera) extract on antioxidant enzymes, performance, digestibility, biochemical parameters and immunity of Egyptian Fayoumi laying chickens. Eur. J. Adva. Res. Biol. Life Sci., 6: 28-40.

Mradu G, Saumyakanti S, Sohini M, Arup M (2012). HPLC profiles of standard phenolic compounds present in medicinal plants. Int. J. Pharm. Pharm. Sci., 4: 162-167.

Nowaczewski S, Babuszkiewicz M, Szablewski T, Stuper-Szablewska K, Cegielska-Radziejewska R, Kaczmarek S, Hejdysz M (2022). Effect of weight and storage time of broiler breeders eggs on morphology and biochemical features of eggs, embryogenesis, hatchability, and chick quality. Animal, 16: 100564. https://doi.org/10.1016/j.animal.2022.100564

Okai MA, Kruenti F, Hamidu JA, Tona K, Hai L (2025). Relationship between in ovo feeding and eggshell temperature of breeder eggs during incubation. Front. Anim. Sci., 6: 1511023. https://doi.org/10.3389/fanim.2025.1511023

Oke OE, Oyelola OB, Iyasere OS, Njoku CP, Oso AO, Oso OM, Daramola JO (2021). In ovo injection of black cumin (Nigella sativa) extract on hatching and post hatch performance of thermally challenged broiler chickens during incubation. Poult. Sci., 100: 100831. https://doi.org/10.1016/j.psj.2020.10.072

Refaie AM, Abd El-Maged MH, Abd El-Halim HH, Alghonimy HR, Shaban SA (2022). Performance, physiological parameters and egg quality traits of laying hens as affected by dietary supplementation of date palm pollen. J. Anim. Health Prod., 10: 21-28. https://doi.org/10.17582/journal.jahp/2022/10.1.21.28

Salhi S, Chentouf M, Harrak H, Rahim A, Çakir C, Çam D, El Amiri B (2024). Assessment of physicochemical parameters, bioactive compounds, biological activities, and nutritional value of the most two commercialized pollen types of date palm (Phoenix dactylifera L.) in Morocco. Food Sci. Technol. Int., 30: 788-798. https://doi.org/10.1177/10820132231168914

Salman KA, Al-Saeedi MK, Al-Jebory HH (2025). Effect of adding manganese sulfate for diet on the hormonal and histological response of aged roosters exposed to heat stress. Kufa J. Agric. Sci., 17: 131-141. https://doi.org/10.36077/kjas/2025/v17i1.12455

SAS (2013). Institute. SAS User’s Guide: Statistics, Version 9.4. Cary (NC): SAS Institute Inc.

Tona K, K. Voemesse, O. N’nanlé, O. Oke, Y. Kouame, A. Bilalissi and O. Oso. 2022. Chicken incubation conditions: role in embryo development, physiology and adaptation to the post-hatch environment. Frontiers in Physiology, 13, 895854. https://doi.org/10.3389/fphys.2022. 895854

Ulmer-Franco AM, Fasenko GM, O’Dea EE (2010). Hatching egg characteristics, chick quality, and broiler performance at two breeder flock ages and from three egg weights. Poult. Sci., 89: 2735–2742. https://doi.org/10.3382/ps.2009-00403

Yalcin S, Özkan S, Shah T (2022). Incubation temperature and lighting: Effect on embryonic development, post-hatch growth, and adaptive response. Front. Physiol., 13: 899977. https://doi.org/10.3389/fphys.2022.899977

Yousaf A, Yasoob TB, Bilal M, Anwar DM, Ghaffar A, Lodhi MK, Malik M (2024). Effect of egg weight on water loss and chicks yield of broiler breeders. Mathews J. Vet. Sci., 8: 1-5. https://doi.org/10.30654/MJVS.10043