Indian Journal of Animal Research

  • Chief EditorM. R. Saseendranath

  • Print ISSN 0367-6722

  • Online ISSN 0976-0555

  • NAAS Rating 6.40

  • SJR 0.233, CiteScore: 0.606

  • Impact Factor 0.4 (2024)

Frequency :
Monthly (January, February, March, April, May, June, July, August, September, October, November and December)
Indexing Services :
Science Citation Index Expanded, BIOSIS Preview, ISI Citation Index, Biological Abstracts, Scopus, AGRICOLA, Google Scholar, CrossRef, CAB Abstracting Journals, Chemical Abstracts, Indian Science Abstracts, EBSCO Indexing Services, Index Copernicus
Submit

Full Research Article

Effects of Pomegranate Peel Polyphenols on Egg Production and Health in Laying Hens

Xu Hongrui1,*, Anqun Huang1, Liangxing Guo1, Furong Nie1, Jianan Hou1, Lv Zengxin1, Mengyan Dang1
1College of Animal Science and Technology, Henan University of Animal Husbandry and Economy, Zhengzhou, Henan Province, 450 046, China.

ABSTRACT

Background: Pomegranate peel polyphenols (PPPs) is a by-product of pomegranate. It has many biological activities, but it has not been used in laying hens. The effect of pomegranate peel polyphenols on laying hens is unclear.

Methods: To understand the role of pomegranate peel polyphenols. We added different concentrations of pomegranate peel polyphenols to the diets of laying hens and studied the changes of growth performance, productivity performance and intestinal microbe during feeding.

Result: The PPPs were found to reduce the feed-to-egg ratio in laying hens. Medium (MP, 500 mg) and high (HP, 750 mg) concentrations of PPPs increased the apparent digestibility of crude protein and calcium. Compared to the control group (CON), all egg performances in the MP group showed significantly improvement. There was a significant increase in reproductive hormones in the MP group compared with the low concentration (LP, 250 mg) and CON groups. As the concentration of PPPs increased, lg Y, SOD and CAT activity increased. The PPPs increased Thyroxine levels significantly. In the LP and MP groups, TNF-a and IL-6 levels were significantly lower than in the CON and HP groups. The relative abundance of Firmcutes increased with the rise in PPPs addition. While Bacteroidota decreased with increasing PPPs concentration. It was concluded that PPPs had a promoting effect on the growth performance for laying hens. This study found the best effects of PPPs addition at the medium concentration because high concentrations would have adverse effects on laying hens’ growth.

INTRODUCTION

Feed additives have been essential substances for the growth process of animals. Suitable feed additives have a significant effect on animal growth and disease resistance. Antibiotics used to be listed as feed additives in China and played an essential role in helping animals fight diseases (Yu et al., 2018). China used to be a significant producer and consumer of antibiotics, but the misuse of antibiotics has adversely affected food safety in the farming industry (Hu et al., 2018). Antibiotics are currently banned for use in farming industry in China (Li et al., 2021), which is a massive challenge for the farming industry.

Plant extracts are environmentally friendly, safe, natural products with good biological activity and low toxicity (Zhou et al., 2009). Studies have shown that plant extracts have the potential to replace antibiotics (Ding et al., 2023) and have demonstrated promising results in improving animal immunity. Meanwhile, plant extracts also exhibit some advantages that antibiotics lack. Most plant extracts have shown sound growth-promoting effects (Lu et al., 2022; Raissy et al., 2022), attributed to their pro-feeding and pro-nutrient uptake abilities. Polyphenol is one of the essential plant extracts that has demonstrated good antioxidant capacity in vitro studies (Zhang et al., 2022) due to its unique polyhydroxy structure. Several plant polyphenols have shown growth-promoting properties in animal production. Adding 2.34 g/kg of grape polyphenols to the diet increased chicken tocopherol concentrations and meat oxidative stability without impairing growth performance (Romero et al., 2021). The addition of 350 or 500 mg/kg of resveratrol, an antioxidant polyphenolic compound found in plants, improved broiler growth performance under heat stress (He et al., 2019). Over 8,000 polyphenols have been identified, but only a few are used for animal production.

Pomegranate peel is one of the essential by-products of pomegranate. Pomegranate peel extracts have multiple biofunctional activities (Peršurić et al., 2020; Botteri et al., 2022; Mo et al., 2022), which are beneficial to human health and have potential applications in the fields of food, medicine, hygiene and nutraceuticals. Polyphenols are one of the main functional components of pomegranate peel (Gigliobianco et al., 2022) and pomegranate peel polyphenols (PPPs) can be obtained by solvent extraction, ultrasound-assisted extraction and microwave-assisted extraction (Nayak et al., 2015). China is currently the world’s largest producer of pomegranates, with the country’s total output in 2019 alone already reaching 1 million tons. The PPPs have high potential value, but few studies have been conducted on using PPPs in animal production. There is insufficient research to support PPPs as a safe feed additive. This research aimed to investigate the utilization of PPPs as feed additives in the diets of laying hens, assess their effect on the growth performance and microbiota and expedite the integration of PPPs into animal production practices.

MATERIALS AND METHODS

Preparation of polyphenols extract
 
The extraction method of pomegranate peel polyphenols was modified with reference to the method of Adithya Sridhar and Takahiro Tsujita (Adithya et al., 2021; Takahiro et al., 2011). The pomegranate peel was dried and ground using a pulverizer, then passed through a 40-mesh sieve. The powder was mixed with 30% ethanol at a solid-to-liquid ratio of 1:40. The mixture was stirred at 70oC for 2 h and the supernatant was collected after centrifugation. The residue was then re-mixed with 30% ethanol at the same solid-to-liquid ratio (1:20) and stirred for an additional 2 h. The supernatant from this second centrifugation was combined with the first supernatant. The concentrate was collected and freeze-dried to obtain pomegranate peel polyphenols. Under these conditions, the total phenol content was (232.016±3.15) mg GAE/g.
 
Experimental design
 
The experiment was conducted with 300 Jing Hong laying hens, 310 d of age, healthy and in the same body condition. The hens were divided into four groups of 75. There were 5 replicates per group and 15 hens per replicate.

The CON group (CON) were fed the normal diet, while experimental groups received additional low (LP, 250 mg), medium (MP, 500 mg) and high concentration (HP, 750 mg) of PPPs, respectively. The total experimental period was seven weeks, including a pre-test period of one week and a formal test period of six weeks. The basal diet was fed during the pre-test period and different concentrations of PPPs were added to the experiment groups during the formal test period.
 
Feeding management and diet formulation
 
Animal testing was conducted by Henan University of Animal Husbandry and Economy in 2023. The feeding experiments were carried out in the egg farm of Henan De An Agricultural. The laying hens in this experiment were raised in cages with water and feed ad libitum. The experimental house was equipped with a 4-layer stepped cage, with 72 cages in each layer and 3 hens in each cage and the feed was fed four times a day. The immunization procedures were carried out in accordance with the daily management of the farm. The daily feeding and management of laying hens was carried out in accordance with the procedures of commercial laying hens. The temperature, humidity, light and hygiene in the chicken house were in accordance with the agricultural industry standard of the People’s Republic of China for experimental animals-environmental and facility standards: GB/T14925-1994 hygienic requirements for egg-laying hen rearing. The composition and nutritional levels of the basal diets are shown in Table 1.

Table 1: Composition and nutrient levels of the basal diet (Dry basis).


 
Production performance
 
Record egg production at 9:00 a.m. daily and calculate egg production rate. The average daily feed intake (ADFI) and Feed-to-Egg (F/E) ratio of laying hens was calculated by counting their feed consumption, recycling and weighing leftover feed each week. Average egg weight calculated by dividing the total egg weight by the number of eggs. Before the end of the formal trial period, each laying hens was weighed once again.
 
Egg quality measurement
 
Eggs from the last three days of the formal trial period were collected. From each replicate, five eggs were randomly selected for egg quality determination. A multifunctional egg quality analyzer determined Haugh Unit and egg white height. Eggshell strength was determined by an eggshell strength tester (KQ-1A, Nannong Animal Husbandry Technology Co. Ltd.) and eggshell thickness, egg shape index and yolk index were determined by vernier caliper.
 
Determination of apparent digestibility
 
The metabolic experiment was conducted for 3 d before the end of the formal trial period. Three healthy laying hens were selected for each replication and clean fecal collection trays were placed under the cage. Fecal samples were collected using the total collection method (Oh et al.,  2020). Proteins were fixed by adding 10% sulfuric acid solution immediately after collection.

The fecal and feed samples were dried at 105oC by oven and crushed through a 0.5 mm mesh screen and nutrients were measured on the feed and fecal samples to calculate the apparent digestibility.
 
  
  
Reproductive hormones and thyroxine estimation
 
At the end of the of the formal trial period, one hen from each replicate was randomly selected for blood collection and serum isolation to determine the levels of reproductive hormones and thyroxine, egg yolk immunoglobulin (IgY), estradiol (E2), luteinizing hormone (LH), triiodothyronine (T3) and thyroxine (T4), follicle stimulating hormone (FSH), were determined by ELISA kits (Shanghai, Enzyme-linked Biotechnology Co., Ltd).
 
Estimation of serum immunity indices
 
Levels of IL-6, TNF-α, lgA and lg M, which were all determined by ELISA kits (Shanghai, Enzyme-linked Biotechnology Co., Ltd).
 
Estimation of serum antioxidant indices
 
The GSH-px, CAT, SOD and MDA levels in serum were determined by commercial kits (Beijing, Solarbio Science  and Technology Co., Ltd).

Serum enzyme activity and calcium estimation
 
Serum aspartate aminotransferase (AST), alanine aminotransferase (ALT) and alkaline phosphatase (AKP) were measured by commercial kits (Beijing, Solarbio Science  and Technology Co., Ltd).

Serum calcium was determined by the methyl thymol blue colorimetric method (Yan et al., 2022).
 
Microbial sequencing analysis
 
At the end of the formal trial period, one hen was randomly selected from each replicate for slaughtering. Colon contents were placed in sterile 5 mL freezing tubes, rapidly cooled with liquid nitrogen. Subsequently stored in a refrigerator at a temperature of -80oC. The colon contents were sent to Wuhan Bena Technology Co. for microbial sequencing.
 
Data analysis
 
Statistical analysis of data was performed using one-way ANOVA by SPSS 16.0 software. A P<0.05 was considered a significant difference. Graphics were drawn using Origin 8.0 software. Graphics for microbiological analysis were plotted using R 3.5.1 software.

RESULTS AND DISCUSSION

Effect of PPPs on growth performance
 
The addition of PPPs significantly reduced the ratio of F/E (P<0.05, Fig 1B). In the MP group, egg production was significantly higher than in the CON group (P<0.05, Fig 1C). Meanwhile, in the LP and HP groups, the final body weight (FBW) was significantly higher (P<0.05, Fig 1D).

Fig 1: Effect of PPPs on the growth performance.


 
Effect of PPPs on apparent digestibility
 
The PPPs significantly increased the apparent digestibility of crude protein (CP, P<0.05, Fig 2C). Additionally, the apparent digestibility of dry matter (DM) and calcium were significantly higher in the MP and HP groups compared to the CON and LP groups (P<0.05, Fig 2A, B).
 

Fig 2: Effect of PPPs on apparent digestibility.



Effect of PPPs on egg quality
 
The albumen height of the MP group was significantly higher than that of the LP and CON groups (P<0.05, Fig 3A). Additionally, compared to the HP and CON groups, the MP group had significantly higher egg weights (P<0.05, Fig 3B). Furthermore, the MP group exhibited significantly higher eggshell strength and thickness than the other experimental groups (P<0.05, Fig 3D, E). Moreover, the yolk height and haugh unit were significantly higher in the PPPs group than in the CON group (P<0.05, Fig 3C, F).
 

Fig 3: Effect of PPPs on egg quality.



Effect of PPPs on serum enzyme activities and calcium levels
 
The figure shows that the MP group had significantly lower AKP levels than the other groups (P<0.05, Fig 4A). In contrast, the LP group had the highest AKP level (P<0.05, Fig 4A). Additionally, in the MP group, serum calcium levels were significantly higher than in the other groups (P<0.05, Fig 4B). Moreover, compared to the CON group, the LP and HP groups had significantly higher serum calcium levels (P<0.05, Fig 4B). Furthermore, serum ALT activity decreased with the addition of increasing PPPs and the serum AST activity was significantly higher in the MP group (P<0.05, Fig 4C, D).

Fig 4: Effect of PPPs on serum enzyme activities and calcium levels.


 
Effect of PPPs on serum reproductive hormones
 
The figure illustrates that serum E2 was significantly higher in the MP group than in the other groups (P<0.05, Fig 5A). Additionally, FSH activity was significantly higher in the MP and HP groups than in the LP and CON groups (P<0.05, Fig 5B). Furthermore, LH levels in the LP and MP groups were significantly higher than in the other two groups (P<0.05, Fig 5C). However, lgY levels in the LP and MP groups were significantly lower than in the CON and HP groups (P<0.05, Fig 5D).

Fig 5: Effects of PPPs on reproductive hormones.


 
Effect of PPPs on serum thyroxine
 
There was a s significant difference in T3 levels in the MP group compared to the other three experimental groups (P<0.05, Fig 6A). Furthermore, the three PPPs groups had significantly higher T4 levels than the CON group (P<0.05, Fig 6B). Meanwhile, the MP group had significantly lower T4 levels than the LP and HP groups (P<0.05, Fig 6B).

​

Fig 6: Effect of PPPs on thyroxine.


 
Effect of PPPs on serum antioxidant capacity
 
As shown in Fig, CAT activity was significantly lower in the HP group (P<0.05, Fig 7A). Additionally, MDA levels in the MP group were significantly lower than those in the HP and CON groups (P<0.05, Fig 7B). Furthermore, SOD activity in the PPPs group increased with the increase of PPPs addition and the difference between the groups was significant (P<0.05, Fig 7D).

​

Fig 7: Effect of PPPs on serum antioxidant capacity.



Effect of PPPs on serum immunity capacity
 
As depicted in figure, the LP and HP groups exhibited significantly higher levels of IgA than the CON group (P<0.05, Fig 8A). Additionally, when compared to the CON and HP groups, the LP group had a significantly higher level of IgM (P<0.05, Fig 8C). Conversely, IL-6 and TNF-a levels were significantly lower in the LP and MP groups than in the CON and HP groups (P<0.05, Fig 8B, D).

​

Fig 8: Effect of PPPs on serum immune indices.


 
Effect of PPPs on microbial α-diversity
 
The figure illustrates that the LP group had significantly lower ACE index and Chao1 index than the other three groups (P<0.05, Fig 9A, C). The Simpson index was not significantly influenced by PPPs, but the Shannon index was significantly lower in the LP group than in the HP group (P<0.05, Fig 9B, D).

​

Fig 9: Effect of PPPs on microbial á-diversity.


 
Effect of PPPs on microbial b-diversity
 
The Venn figure illustrates similarities in microbial structure among the four groups, with notable differences (Fig 10A). The total number of identical OTUs in the four groups was 618 and the number of OTUs unique to each group increased with the amounts of PPPs added. PCA analysis showed differences among the groups with varying concentrations of PPPs, while still maintaining some similarity with the CON group (Fig 10B).

​

Fig 10: Venn diagram with microbial â-diversity analysis.


 
Effect of pomegranate peel polyphenols on microbial community structure
 
The figure shows that the relative abundance of Firmcutes increased with the rise in PPPs addition (Fig 11A). While Bacteroidota decreased with increasing PPPs concentration. The relative abundance of Fusobacteriota was significantly lower in the MP group than in the other groups. Meanwhile, the relative abundance of Proteobacteria in the MP group was significantly higher than the other groups.

​

Fig 11: Structural composition of microorganisms at phylum level and species level.



Microorganisms significantly affected by PPPs can be more precisely identified at the species level (Fig 11B). The relative abundance of Megamonas_hypermegale, Bacteroides_salanitronis, Bacteroides_gallinaceum and Bacteroides_caecigallinarum in the MP group was significantly higher than the other groups. The relative abundance of Desulfovibrio_sp and Bacteroides_plebeius was significantly lower in the MP group than in the other groups. The relative abundance of Firmicutes_bacterium and Clostridium_spiroforme increased with increasing PPPs concentrations.
 
Microbiological correlation analysis
 
Spearman’s correlation coefficient was used to analyse the correlation between microbial species levels and to generate heat maps. Fig 12 shows a significant correlation between antioxidant capacity and immuno competence and microorganisms in laying hens. There was a significant negative correlation between Alloprevotella, Anaerotruncus, Candidatus vestibaculum, Succinatimonas and IL6. The correlation between Anaerostipes and SOD, Ig A, Ig M was significantly negative but with Ig Y it was significantly positive. Between Barnesiella and MDA, TNF-α, Ig Y and IL6 is a significant positive correlation. A strong link can be seen between microorganisms and antioxidant and immune competence.

​

Fig 12: Correlation analysis of microorganisms with immune and antioxidant indicators.



Plant polyphenols, as essential active plant extracts, are known to promote animal growth performance (Singh et al., 2021; Chen et al., 2024; Rao et al., 2023). In the present study, the laying rate and final body weight of laying hens in the PPPs group were enhanced compared to the CON group. However, there was no significant difference in feed intake among the groups. Studies have shown that plant polyphenols can improve feed utilization in poultry (Shakeri et al., 2020; Qin et al., 2024). Therefore, we determined the apparent digestibility and found that the PPPs group had improved the apparent digestibility of CP and DM, consistent with previous studies. This means that PPPs promote nutrient uptake in laying hens to improve growth performance.

Meanwhile, we found that the apparent digestibility of calcium was significantly higher in the MP and HP group than in the CON group. Calcium is eggshells’ main building block and significantly affects eggshell strength and thickness (Vijay et al., 2021). The increased serum calcium levels and apparent digestibility of calcium in laying hens in the PPPs group indicated that PPPs promoted calcium absorption in laying hens. This was confirmed by significantly higher eggshell strength and thickness in the MP group than the CON. The PPPs improved both egg white height, yolk height and haugh unit, with the best results obtained with MP group, which may be related to improved apparent digestibility of nutrients.

As an essential defense organ of the organism, the liver is resistant to exogenous additives, which may produce liver damage in animals (Ye et al., 2018). Liver damage releases large amounts of metabolites, such as ALT, into the bloodstream (Shao et al.,  2022). Changes in metabolites such as ALT and AST in the blood can directly reflect liver damage. A significant decrease in ALT levels observed in three PPPs groups compared with the CON group. This suggests that PPPs have the ability to protect the liver.

Reproductive hormones (E2, FSH and LH) in laying hens directly impact egg production and F/E ratio (He et al., 2022). The significant improvement in F/E ratio  and egg production rate of the PPPs group might be attributed to increased E2, FSH and LH levels of serum in the PPP groups, indicating improved reproductive performance of the laying hens. However, studies have shown that excessive polyphenol concentrations can instead impair reproductive performance (Fraser, 2008). Similarly, in present study also levels of reproduction hormones and corresponding egg production performance, decreased at higher supplementation rate in HP group above medium level of supplementation in MP groups. Thyroid hormones are mainly involved in the body’s material metabolism processes (Rafique et al., 2020). T4 is the main secretion product of the thyroid gland, which is relatively inactive and is converted into the highly active T3 by 52  deiodinase (Mancini et al., 2005). Studies have shown that serum T3 decreases under stress. (Meng et al., 2022). In contrast, the addition of thyroxine to the diet improves egg production in laying hens, which is attributed to the fact that thyroid hormones enhance the body’s metabolism and promote the absorption and utilization of glucose by the cells. (Yehuda-Shnaidman et al.,  2014; Rastogi et al., 2015). The organism’s enhanced nutrient utilization efficiency leads to an increase in production performance. In the present study, we found that PPPs increased serum T3 and T4 levels, which helped to improve nutrient utilization in laying hens, in line with our previous findings of increased apparent digestibility of CP and DM.

The most crucial function of plant polyphenols is improving the body’s antioxidant capacity and immunity. (Taher et al., 2022; Liao et al., 2019; Khan et al., 2019). Several studies have confirmed that plant polyphenols exhibit good antioxidant capacity in vivo and in vitro. Immunity and antioxidant capacity are closely related (Liu et al., 2019). Plant polyphenols indirectly enhance immunity by increasing antioxidant capacity. PPPs showed similar effects in the present study. PPPs increased serum SOD activity and decreased MDA levels. However, CAT activity was significantly lower in the HP group than in the CON group, which suggests that high concentration of PPPs may have side effects on the organism. Changes in serum immunocompetence in the PPPs group were similar to antioxidant capacity. Low and medium concentrations of PPPs reduced the levels of TNF-á, but high concentration of PPPs did not show similar results. Meanwhile, the LP and MP groups reduced the levels of pro-inflammatory factor IL-6. The HP group were not significantly different from the CON group, however. The high concentration PPPs group had significantly higher levels of lg A than the CON group, but the lg M level did not differ significantly. The above results indicate that PPPs can improve antioxidant capacity and immunity in laying hens. There was a significant correlation with the concentration of PPPs.

The colonic microorganisms of laying hens have significant effects on nutrient absorption and immune performance (Xie et al., 2020). Many studies have shown that plant polyphenols can significantly affect the structure of intestinal microbial communities (Scarsella et al., 2020; Sweeney et al., 2022). Polyphenols are only partially degraded in the stomach; the vast majority can enter the intestinal tract to work. (Jakesevic et al., 2011; Han et al., 2020). Therefore, polyphenols can indirectly alter laying hens’ nutrient uptake and immunity by influencing the structure of the colonic microbial community. β-diversity structure showed that PPPs influenced the colonic microbial community structure of laying hens and that low of PPPs reduced the microbial community abundance. β-diversity structure also showed that PPPs influenced the microbial community structure and there was a difference between the groups of different PPPs concentrations. Firmicutes play a crucial role in nutrition and metabolism through short chain fatty acid synthesis (Jin et al., 2024). PPPs significantly up-regulated the relative abundance of Firmicutes. PPPs also inhibit the growth of harmful bacteria, including Fusobacteria, Campylobacter and Fusobacteria_ mortiferum. Meanwhile, PPPs also inhibit the growth of some bacteria that utilize nutrients, including Halobacterota, to achieve a high efficiency of nutrient utilization in the intestinal tract, especially the utilization of amino acids. While inhibiting harmful bacteria, PPPs promoted the growth of some probiotics, especially at MP group, where the most significant performance was observed. Megamonas hypermegale can ferment various carbohydrates (Shang et al., 2020), with the end products being acetic, propionic and lactic acids. Intestinal epithelial cells will eventually absorb short chain fatty acids, enhancing the body’s immunity (Ren et al., 2022). Medium concentration of PPPs promoted the growth of this bacterium, but high concentration of PPPs, on the contrary, did not show promotive results. The heat map also showed that the structure of the colonic microbial community is closely related to antioxidant properties and immunity. This is because various metabolites of microorganisms can enter the bloodstream through the intestinal barrier, directly affecting the organism’s antioxidant and immune properties. Therefore, PPPs can inhibit the growth of harmful bacteria and promote the growth of intestinal probiotics. The change is closely related to the effect of PPPs addition.

CONCLUSION

The addition of PPPs to the diets of laying hens improved gut health by regulating the gut microbial community. Additionally, PPPs increased antioxidant capacity and immune function by promoting probiotics and inhibiting harmful bacteria. This, in turn, enhanced nutrient utilization, particularly the absorption of dry matter (DM), crude protein (CP) and calcium, which positively impacted metabolic and immune status. The improved nutrient absorption contributed to better reproductive hormone levels, promoting increased egg production and improved egg quality. However, the effect of PPPs did not increase with the rise of concentration and the high concentration of PPPs did not perform significantly more potent than the medium concentration of PPPs in all aspects, especially the levels of the LH and E2, which were instead reduced. Specifically, PPPs enhanced eggshell strength and thickness through better calcium utilization. Therefore, the optimal concentration of PPPs in this study was medium. PPPs have good prospects for application as natural extract additives.

ACKNOWLEDGEMENT

This present study was supported by the National Key Research and Development Program of China (2018YFD0501103).
 
Disclaimers
 
The views and conclusions expressed in this article are solely those of the authors and do not necessarily represent the views of their affiliated institutions. The authors are responsible for the accuracy and completeness of the information provided, but do not accept any liability for any direct or indirect losses resulting from the use of this content.
 
Informed consent
 
The experiment was conducted in accordance with the Guide for Care and Use of Laboratory Animals of the National Research Council, China (GB/T 35892-2018). The experiment was approved and supervised by the Experimental Animal Ethics Committee of Henan University of Animal Husbandry and Economy.
 
Authors contribution
 
Hongrui Xu: Conceptualization, Methodology, Writing, review  and editing, Supervision. Zengxin Lv, Mengyan Dang: Writing original draft, Experimental operation. Anqun Huang, Liangxin Guo, Furong Nie, Jianan Hou: Methodology, Writing, review  and editing. All authors have read, improved and approved the manuscript.
 
Data availability
 
Data will be made available on request.

CONFLICT OF INTEREST

The authors declare that there are no conflicts of interest.

REFERENCES


  1. Adithya, S., Muthamilselvi, P., Senthil, P.K., Ashish, K., Dai-Viet, N.V., Sivaraman P. (2021). Techniques and modeling of polyphenol extraction from food: A review. Environmental Chemistry Letters. 19(4): 31-35.

  2. Botteri, L., Miljkovic, A., Glogar, M.I. (2022). Influence of cotton pre-treatment on dyeing with onion and pomegranate peel extracts. Molecules. 27(14): 4547.

  3. Chen, A., Ma, T., Zhong, Y., Deng, S., Zhu, S., Fu, Z., Huang, Y., Fu, J. (2024). Effect of tea polyphenols supplement on growth performance, antioxidation and gut microbiota in squabs. Frontiers in Microbiology. 14: 1329036.

  4. Ding, X., Giannenas, I., Skoufos, I., Wang, J., Zhu, W. (2023). The effects of plant extracts on lipid metabolism of chickens - A review. Animal Bioscience. 36(5): 679-691.

  5. Fraser. L.R. (2008). The role of small molecules in sperm capacitation. Theriogenology. 70(8): 1350-1359.

  6. Gigliobianco, M.R., Cortese, M., Nannini, S., Di Nicolantonio, L., Peregrina, D.V., Lupidi, G., Vitali, L.A., Bocchietto, E., Di Martino, P., Censi, R. (2022). Chemical, antioxidant and antimicrobial properties of the peel and male flower by- products of four varieties of Punica granatum L. cultivated in the marche region for their use in cosmetic products. Antioxidants. 11(4): 768. 

  7. Han, Y., Xiao, H. (2020). Whole food-based approaches to modulating gut microbiota and associated diseases. Annual Review of Food Science and Technology. 11: 119-143.

  8. He, H., Li, D., Tian, Y., Wei, Q., Amevor, F.K., Sun, C., Yu, C., Yang, C., Du, H., Jiang, X., Ma, M., Cui, C., Zhang, Z., Tian, K., Zhang, Y., Zhu, Q., Yin, H. (2022). miRNA sequencing analysis of healthy and atretic follicles of chickens revealed that miR-30a-5p inhibits granulosa cell death via targeting Beclin1. Journal of Animal Science and Biotechnology. 13(1): 55.

  9. He, S., Li, S., Arowolo, M.A., Yu, Q., Chen, F., Hu, R., He, J. (2019). Effect of resveratrol on growth performance, rectal temperature and serum parameters of yellow-feather broilers under heat stress. Animal Science Journal. 90(3): 401-411. 

  10. Hu, J., Ma, L., Nie, Y., Chen, J., Zheng, W., Wang, X., Xie, C., Zheng, Z., Wang, Z., Yang, T., Shi, M., Chen, L., Hou, Q., Niu, Y., Xu, X., Zhu, Y., Zhang, Y., Wei, H., Yan, X. (2018). A microbiota-derived bacteriocin targets the host to confer diarrhea resistance in early-weaned piglets. Cell Host  and Microbe. 24(6): 817-832.e8.

  11. Jakesevic, M., Aaby, K., Borge, G.I., Jeppsson, B., Ahrné, S., Molin, G. (2011). Antioxidative protection of dietary bilberry, chokeberry and Lactobacillus plantarum HEAL19 in mice subjected to intestinal oxidative stress by ischemia- reperfusion. BMC Complementary and Alternative Medicine. 11: 8. https://doi.org/10.1186/1472-6882-11-8.

  12. Jin, S.W., Han, Y.C., Sun, Y.G., Ding, W.Q., Liu, Y.Q., Gou, F.J., Chen, J.Y. (2024). Comparative analysis of rumen fermentation and microbial communities in yaks of different ages. Indian Journal of Animal Research. 12. doi: 10.18805/ IJAR.BF-1873.

  13. Khan, H., Sureda, A., Belwal, T., Çetinkaya, S., Süntar, Ý., Tejada, S., Devkota, H.P., Ullah, H., Aschner, M., (2019). Polyphenols in the treatment of autoimmune diseases. Autoimmunity Reviews. 18(7): 647-657. 

  14. Li, Y., Wang, Y., Lv, J., Dou, X., Zhang, Y., (2021). Effects of dietary supplementation with clostridium butyricum on the amelioration of growth performance, rumen fermentation and rumen microbiota of holstein heifers. Frontiers in Nutrition. 8: 763700. 

  15. Liu, X., Wan, T., Dang, S., Wang, D., Jiang, C., Su, Y., Shen, M., Tang, X., Li, X., Li, B., Wen, L., Feng, Q. (2019). HLA- DQB1/DRB1 alleles associate with traditional chinese medicine syndrome of chronic hepatitis b: A potential predictor of progression. Biomed Research International. 2019: 8146937. 

  16. Liao, J.L., Shen, X.M., Yu, S.G., Wang. G., Long, W.C., Wang, Y., Yang, W.K.Y., Peng, F., Fan, Y.Z., Liang, Z. (2024). Effects of allicin on the immune performance and expression of immune-related genes in muchuan black-bone chickens. Indian Journal of Animal Research. 58(7): 1088-1095.

  17. doi: 10.18805/IJAR.BF-1795.

  18. Lu, Y., Zhao, M., Mo, J., Lan, G., Liang, J. (2022). Dietary supplementation ellagic acid on the growth, intestinal immune response, microbiota and inflammation in weaned piglets. Frontiers in Veterinary Science. 9: 980271. 

  19. Mancini, A., Corbo, G.M., Gaballo, A., Valente, S., Gigliotti, P., Cimino, V., De Marinis, L., Principi, F., Littarru, G.P. (2005). Relationships between plasma CoQ10 levels and thyroid hormones in chronic obstructive pulmonary disease. BioFactors. 25(1-4): 201-204.

  20. Meng,T., Deng, J., Xiao, D., Arowolo, M.A., Liu, C., Chen, L., Deng, W., He, S., He, J. (2022). Protective effects and potential mechanisms of dietary resveratrol supplementation on the spleen of broilers under heat stress. Frontiers in Nutrition. 9: 821272.

  21. Mo, Y., Ma, J., Gao, W., Zhang, L., Li, J., Li, J., Zang, J. (2022). Pomegranate peel as a source of bioactive compounds: a mini review on their physiological functions. Frontiers in Nutrition. 9: 887113. 

  22. Nayak, B., Dahmoune, F., Moussi, K., Remini, H., Dairi, S., Aoun, O., Khodir, M. (2015). Comparison of microwave, ultrasound and accelerated-assisted solvent extraction for recovery of polyphenols from citrus sinensis peels. Food Chemistry. 187: 507-16. 

  23. Oh, H.J., Yun, W., Lee, J.H., An, J.S., Kim, T.H., Cho, E.A., Park, B.J., Kim, K.H., Lee, S.D., Cho, J.H. (2020). Effect of replacing corn with soy hulls on nutrient digestibility of growing pigs. Journal of Animal Science and Technology. 62(2): 180-186.

  24. Peršuric, Ž., Saftic Martinovic, L., Malenica, M., Gobin, I., Pedisic, S., Dragovic-Uzelac, V., Kraljeviæ Pavelic, S. (2020). Assessment of the biological activity and phenolic composition of ethanol extracts of pomegranate (Punica granatum L.) peels. Molecules. 25(24): 5916. 

  25. Qin, M., Wang, Z., Liang, M., Sha, Y., Liu, M., Liu, J., Wang, T., Zhao, C., Wang, Z., Guo, D., Li, R. (2024). Effects of dietary supplementation with tea polyphenols and probiotics on laying performance, biochemical parameters intestinal morphology and microflora of laying hens. International Journal of Biological Macromolecules. 256: 128368.

  26. Rafique, O., Mir, A.H. (2020). A topological approach for cancer subtyping from gene expression data. Journal of Biomedical Informatics. 102: 103357. 

  27. Raissy, M., Ahmadi Kabootarkhani, M., Sanisales, K., Mohammadi, M., Rashidian, G. (2022). The synergistic effects of combined use of mentha longifolia, thymus carmanicus and trachyspermum copticum on growth performance, feed utilization and expression of key immune genes in rainbow trout (Oncorhynchus mykiss). Frontiers in Veterinary Science. 8: 810261.

  28. Rao, Z.X., Tokach, M.D., Woodworth, J.C., DeRouchey, J.M., Goodband, R.D., Shah, A.S., Foley, B.H., Kjeldsen, K.C., Brunsgaard, G., Gebhardt, J.T. (2023). Polyphenols as a partial replacement for vitamin E in nursery pig diets. Translational Animal Science. 7(1): txad116.

  29. Rastogi, D., Fraser, S., Oh, J., Huber, A.M., Schulman, Y., Bhagtani, R.H., Khan, Z.S., Tesfa, L., Hall, C.B., Macian, F. (2015). Inflammation, metabolic dysregulation and pulmonary function among obese urban adolescents with asthma. American Journal of Respiratory and Critical Care Medicine. 191(2): 149-60. 

  30. Ren, D.D., Li, S.S., Lin, H.M., Xia, Y.S., Li, Z.M., Bo, P.P., Mu, R., Zhao, L.J., Sun, Y.S. (2022). Panax quinquefolius polysaccharides ameliorate antibiotic-associated diarrhoea induced by lincomycin hydrochloride in rats via the MAPK signaling pathways. Journal of Immunology Research. 2022: 4126273.

  31. Romero, C., Nardoia, M., Arija, I., Viveros, A., Rey, A.I., Prodanov, M., Chamorro, S. (2021). Feeding broiler chickens with grape seed and skin meals to enhance a-and a-tocopherol content and meat oxidative stability. Antioxidants. 10(5): 699.

  32. Scarsella, E., Cintio, M., Iacumin, L., Ginaldi, F., Stefanon, B. (2020). Interplay between neuroendocrine biomarkers and gut microbiota in dogs supplemented with grape proantho- cyanidins: results of dietary intervention study. Animals. 10(3): 531.

  33. Shakeri, M., Cottrell, J.J., Wilkinson, S., Le, H.H., Suleria, H.A.R., Warner, R.D., Dunshea, F.R. (2020). A dietary sugarcane- derived polyphenol mix reduces the negative effects of cyclic heat exposure on growth performance, blood gas status and meat quality in broiler chickens. Animals. 10(7): 1158. 

  34. Shakoor, H., Feehan, J., Apostolopoulos, V., Platat, C., Al Dhaheri, A.S., Ali, H.I., Ismail, L.C., Bosevski, M., Stojanovska, L. (2021). Immunomodulatory effects of dietary polyphenols. Nutrients. 13(3): 728. 

  35. Shang, H., Zhao, J., Dong, X., Guo, Y., Zhang, H., Cheng, J., Zhou, H. (2020). Inulin improves the egg production performance and affects the cecum microbiota of laying hens. International Journal of Biological Macromolecules. 155: 1599-1609.

  36. Shao, H., Xiao, M., Zha, Z., Olatunji, O.J. (2022). UHPLC-ESI-QTOF- MS2 analysis of Acacia pennata extract and its effects on glycemic indices, lipid profile, pancreatic and hepatorenal alterations in nicotinamide/streptozotocin-induced diabetic rats. Food Science and Nutrition. 10(4): 1058-1069. 

  37. Singh, S., Hundal, J.S., Patra, A.K., Sethi, R.S., Sharma, A. (2021). A composite polyphenol-rich extract improved growth performance, ruminal fermentation and immunity, while decreasing methanogenesis and excretion of nitrogen and phosphorus in growing buffaloes. Environmental Science and Pollution Research. 29(17): 24757-24773.

  38. Sweeney, M., Burns, G., Sturgeon, N., Mears, K., Stote, K., Blanton, C. (2022). The effects of berry polyphenols on the gut microbiota and blood pressure: a systematic review of randomized clinical trials in humans. Nutrients. 14(11): 2263.

  39. Takahiro, T., Misato, Y., Takeshi, T., Tomoyoshi, S., Kanae, T., Takafumi, S. (2011). Purification and characterization of polyphenols from chestnut astringent skin. Journal of Agricultural and Food Chemistry. 59(16): 8646-8654.

  40. Taher, M.A., Lo’ay, A.A., Gouda, M., Limam, S.A., Abdelkader, M.F.M., Osman, S.O., Fikry, M., Ali, E.F., Mohamed, S.Y., Khalil, H.A., El-Ansary, D.O., El-Gioushy, S.F., Ghazzawy, H.S., Ibrahim, A.M., Maklad, M.F., Abdein, M.A., Hikal, D.M. (2022). Impacts of gum arabic and polyvinylpyrrolidone (PVP) with salicylic acid on peach fruit (Prunus persica) shelf life. Molecules. 27(8): 2595.

  41. Vijay, A., Gowda, N.K.S., Pal, D.T., S. Debpriyo Kumar Dey., Karthik Bhat, S., Elangovan, A.V. (2021). Boron supplementation to calcium inadequate diet influences mineral content of serum, tissue and antioxidant status in white leghorn layers. Indian Journal of Animal Research. doi: 10.18805/ IJAR.B-4493.

  42. Xie, Y., Wang, C., Zhao, D., Zhou, G., Li, C. (2020). Processing method altered mouse intestinal morphology and microbial composition by affecting digestion of meat proteins. Frontiers in Microbiology. 11: 511.

  43. Yan, Y., Luo, Z., He, L., Wei, F., Gao, M., Cui, X., Yang, J., Sun, Y., Li, L., Yang, L., Deng, T., Pan, X., Peng, M., Tan, Y., An, Z., Li, L., Yang, X. (2022). Preparation and characterization of calcium-incorporated rosa roxburghii tratt and its efficacy on bone mineral density in rats. Evidence-Based Complementary and Alternative Medicine. 2022: 5122396. 

  44. Ye, H., Nelson, L.J., Gómez Del Moral, M., Martínez-Naves, E., Cubero, F.J. (2018). Dissecting the molecular pathophysiology of drug-induced liver injury. World Journal of Gastro- enterology. 24(13): 1373-1385.

  45. Yehuda-Shnaidman, E., Kalderon, B., Bar-Tana, J. (2014). Thyroid hormone, thyromimetics and metabolic efficiency. Endocrine Reviews. 35(1): 35-58.

  46. Yu, D.Y., Kim, S.H., Kim, J.A., Kim, I.S., Moon, Y.S., Lee, S.S., Park, H.C., Jung, J.H., Chung, Y.H., Shin, D.K., Nam, K.C., Choi, I.S., Cho, K.K. (2018). Effects of Rubus coreanus by products on intestinal microbiota and the immune modulation. Asian-Australasian Journal of Animal Sciences. 31: 429- 438.

  47. Zhang, C., Yu, J., Tu, Q., Yan, F., Hu, Z., Zhang, Y., Song, C. (2022). Antioxidant capacities and enzymatic inhibitory effects of different solvent fractions and major flavones from celery seeds produced in different geographic areas in China. Antioxidants. 11(8): 1542. 

  48. Zhou, Q., Li, L., Xiang, J., Sun, H., Tang, Y. (2009. Fast screening and structural elucidation of g-quadruplex ligands from a mixture via g-quadruplex recognition and NMR methods. Biochimie. 91(2): 304-308.
Reviewed By

Download Article
In this Article
    Contact us
    Follow us

    Editorial Board

    View all (0)