
==== Front
Poult Sci
Poult Sci
Poultry Science
0032-5791
1525-3171
Elsevier

S0032-5791(24)00757-0
10.1016/j.psj.2024.104178
104178
METABOLISM AND NUTRITION
Dietary betaine supplementation improved egg quality and gut microbes of laying hens under dexamethasone-induced oxidative stress
Wang Chaohui
Liu Xiaoying
Sun Xi
Li Yun
Yang Xiaojun
Liu Yanli liuyanli@nwsuaf.edu.cn
1
College of Animal Science and Technology, Northwest A&F University, Yangling, 712100, China
1 Corresponding author: liuyanli@nwsuaf.edu.cn
07 8 2024
11 2024
07 8 2024
103 11 10417823 5 2024
1 8 2024
© 2024 The Authors
2024
https://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
Oxidative stress is a frequent concern in the breeding of laying hens, and limit the healthy development of poultry. Dexamethasone (DXM) has been demonstrated to induce oxidative stress. Conversely, betaine is an alkaloid with a potent antioxidant activity. The study was designed to investigate the ameliorative effect of betaine on DXM-induced oxidative stress in laying hens. The results revealed that DXM treatment significantly decreased laying rate, shell strength, albumen height, Haugh unit, egg weight, folk weight and albumen weight, alongside increased malondialdehyde (MDA) and decreased total antioxidant capacity (T-AOC) in serum and liver (P < 0.05). In contrast, dietary betaine addition reversed those parameters mentioned above (P < 0.05). Hepatic RNA-seq analysis showed that there existed 110 up- and 88 down-regulated genes in DXM group when compared with the control. Meanwhile there were 117 upregulation and 169 downregulation genes in BT group when compared with DXM group. Besides, we found that dietary betaine addition significantly down-regulated cell adhesion molecules, glycerolipid metabolism and glycolysis gluconeogenesis pathways. In addition, a total of 44 and 94 differential metabolites were identified respectively from Con vs. DXM and DXM vs BT. More importantly, dietary betaine addition significantly increased the levels of pantothenic acid, gamma-Aminobutyric acid, equol and choline, all of which were related to antioxidant and anti-inflammatory properties. Furthermore, gut microbiota analysis indicated that the Chao and Observed_species indexes were remarkably higher in BT group (P<0.05). Heatmap analysis revealed that Subdoligranulum, Prevotella, Blautia, YRC22, Bacteroides, Ruminococcus and Coprococcus were notably restored in BT group (P<0.05). Taken together, our findings collectively illustrate that dietary betaine addition could attenuate DXM-induced oxidative stress, improve egg quality and gut microbes of laying hens.

Key words

Betaine
egg quality
gut microbe
laying hen
oxidative stress
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pmcINTRODUCTION

In livestock and poultry rearing, various unfavorable factors easily induce stress reactions, encompassing physiological stress, environmental conditioned stress, physical stress, immune stress and disease stress, which can ultimately result in oxidative stress (Surai et al., 2019; Eid et al., 2021). The overreaction of oxidative stress in poultry can significantly decrease feed intake and production performance, thereby causing intestinal inflammation, hindering growth and development (Liu et al., 2022). On the other hand, it was reported that oxidative stress could affect egg quality traits, yolk lipids, and cholesterol contents (Duan et al., 2014; Eid et al., 2021; de Brito et al., 2022). These detrimental effects pose significant threats to poultry health and cause substantial economic losses in poultry industry. Furthermore, studies have demonstrated that oxidative stress-induced hepatic damage contributes to the development and advancement of nonalcoholic steatohepatitis (NASH) (Singal et al., 2011; Feng et al., 2011; Fan et al., 2023). Therefore, it is urgent to find nutritional strategies to prevent or alleviate oxidative stress.

It has been reported that oxidative stress activates the hypothalamo-pituitary-adrenocortical (HPA) axis, leading to the secretion of glucocorticoids (Herman et al., 2016). Elevated levels of glucocorticoids, a hallmark of stress, have been proven to be involved in the alteration of redox balance in poultry (Osho and Adeola, 2020; Mir et al., 2021). Thus, dexamethasone (DXM), a synthetic glucocorticoid with anti-inflammatory and anti-immunosuppressive effects, has been used as a stress models in many studies (Berenjian et al., 2018; Delanogare et al., 2020; Barekatain et al., 2021). Additionally, betaine, a natural compound found abundantly in various foods, was reported to enhance the production performance of broilers (Yusuf et al., 2018), decrease hepatic cholesterol deposition in chicken (Hu et al., 2020) and increase the laying rate of laying hens (Abobaker et al., 2017). What's more, Sun et al. (Kim et al., 2008) reported that betaine can alleviate ethanol-induced oxidative stress in mice by reducing CYP2E1 activity and NOS2 protein levels. Wen et al. (Wen et al., 2019) found that dietary supplementation with betaine could mitigate the negative effects of heat stress on the oxidation status of broilers. Du et al. found (Du et al., 2021) that betaine can regulated lipid metabolism by increasing antiobesity strains such as Akkermansia Muciniphila, Lactobacillus, and Bifidobacterium. However, there is little evidence about how betaine regulate host metabolism. The current study was conducted to evaluate the relieving effect of betaine on DXM-induced oxidative stress through hepatic transcriptional changes, serum metabolomics and gut microbiota analysis.

MATERIALS AND METHODS

Animals

The animals used (Hy-Line Brown laying hens) were all purchased from Julong Poultry Farm (Wugong, Shanxi, China) and the experimental procedures in this study were conducted in accordance with the guidelines and regulations at the Animal Ethics Committee of Northwest A&F University (Permit Number: DK202123).

Experimental Design

A total of 180 Hy-line Brown hens aged 21-wk-old were randomly divided into 3 groups, including the Con group (basal diet), the DXM group (dexamethasone injection + basal diet) and the BT group (extra 8 g/kg betaine based on DXM group). Each group had 10 replicates and 6 birds in each replicate. After 1 wk prefeeding, hens in DXM group and BT group were subcutaneously injected with DXM (4.5 mg/kg body weight) for 7 consecutive d at 8:00 in the morning, and hens in Con group were injected with an equivalent volume of saline at the same time. DXM was obtained from Chen Xin Pharmaceutical Co., Ltd (Shangdong, China). The experimental diet was a corn-soybean-basal diet, formulated according to the guidelines provided by the Chinese Feeding Standard of Chickens (NY/T33-2004). The detailed composition of the basal diet was shown in Table 1. The birds were housed in an environmentally controlled cages (1 bird/cage) (depth × width × height = 45 cm × 35 cm × 45 cm) at the poultry farm of Northwest A&F University. The temperature was maintained within a range of 25±2°C, and the relative humidity was controlled from 60 to 70%. Eggs were collected once daily for calculation of laying rate and egg quality was detected by EMT-7300 egg multi tester (Robotmation, Japan). The duration trial lasted for 6 wk and the experimental design was shown in Figure 1.Table 1 Composition and nutrient levels of basal diet.

Table 1Composition (air-dry basis) %	Basal diet	
Corn	56.69	
DDGS	4.00	
Soybean meal (43%)	25.77	
DL-Methionine	0.18	
Fat-soybean oil	1.51	
CaCO3	9.04	
CaHPO4 21/16	1.15	
NaCl	0.26	
Choline chloride (60%)	0.15	
Premix⁎	1.00	
Bentonite	0.25	
Total	100.00	
Nutrient levels		
Metabolizable energy, kcal/kg (calculated)	2,600	
Crude protein (calculated)	16.5	
Total phosphorus (calculated/analyzed)	0.53/0.49	
Nonphytate phosphorus (calculated)	0.32	
Calcium (calculated/analyzed)	3.50/3.52	
⁎ The composition of premixes: iron, 60 mg; manganese, 60 mg; copper, 8 mg; zinc, 80 mg; selenium, 0.3 mg; iodine, 0.35 mg; vitamin A, 8000 IU; vitamin D3, 1600 IU; vitamin E, 30 mg; menadione, 1.5 mg; vitamin C, 200 mg, thiamine, 4 mg; riboflavin, 13 mg; pantothenic acid, 15 mg; nicotinamide, 20 mg; pyridoxine, 6 mg; biotin, 0.15 mg; folic acid, 1.5 mg; cobalamin, 0.02 mg; additional 8000 mg betaine was added to the betaine group.

Figure 1 Strategy of the experimental design used to evaluate the effects of BT on DXM-induced stress in chickens.

Figure 1

Sample Collection

After 6 wk feeding, 1 bird was randomly selected from each replicate. Approximately 5 mL of blood was collected from wing vein and serum was obtained after centrifugation at 3,000 × g for 10 min. Thereafter hens were sacrificed by cervical dislocation and dissected. The liver and fresh cecum chymus was collected and then transferred to −80°C for further analysis.

Serum Biochemical Examination

Serum biochemical parameters including aspartate aminotransferase (AST), Lactate dehydrogenase (LDH), total bilirubin (TBIL), total protein (TP), total biliary acid (TBA), total cholesterol (TC), triglyceride (TG), high-density lipoprotein cholesterol (HDL-c) and low-density lipoprotein cholesterol (LDL-c) were determined using 7,180 Clinical Analyzer (Hitachi, Japan) at Yangling Demonstration Zone Hospital (Yangling, China). Additionally, Total antioxidant capacity (T-AOC) and Malondialdehyde (MDA) were measured using a commercial kit (Nanjing Jiancheng Institute of Biological Engineering, Nanjing, China) according to the kit instructions.

Antioxidant Function Analysis in the Liver

The liver tissues were homogenized with cold PBS by high-throughput homogenizer and the supernatant were collected after centrifugation at 3,000 × g for 10 min at 4°C. The protein concentration was detected by BCA commercial kit (Xi'an AccuRef Scientific Co., Ltd, Xi'an, China). Additionally, T-AOC and MDA contents were measured through commercial kits (Nanjing Jiancheng Institute of Biological Engineering, Nanjing, China). Data conversion and standardization were performed by protein concentration.

Transcriptomics Analysis

The total RNA in the liver was extracted by using Trizol reagent (Accurate Biotechnology Co., Ltd, Changsha, China), after which the concentration and quality were determined using a NanoDrop spectrophotometer (Thermo Fisher Scientific, Shanghai, China). The library construction and sequencing were performed at by Shanghai Personal Biotechnology Co., Ltd through llumina NovaSeq. The sequencing data was filtered to remove the low-quality reading with fastp (v0.22.0) software, and then the filtered reads were aligned to the reference genome (GRCg7b, GCF_016699485.2, https://www.ncbi.nlm.nih.gov/genome/111) using HISAT2 (http://ccb.jhu.edu/software/hisat2/index.shtml) software. The differential expression genes (DEG) were identified according to the parameters of log2 fold change > 1 and P-value < 0.05. The volcano map and clustering heatmap of DEGs were constructed by using R language ggplots2 and R language Pheatmap (v1.0.12) package. Gene Set Enrichment Analysis (GSEA) was performed using cluster profiler, and the significant enrichment criterion was |NES|>1, P-value < 0.05, FDR < 0.25.

Metabolomics Analysis

Serum metabolites extraction was following the method described previously (Demurtas et al., 2021). Untargeted serum metabolomics was performed using an GC–MS platform at Shanghai Personal Biotechnology Co., Ltd. GC–MS data selected, aligned and a series of processing based on the previous reports (He et al., 2023). All multivariate data analysis and modeling were performed using Ropls software, and the data was centered on the mean value by scaling. The repository names and accession numbers for metabolomics is www.ebi.ac.uk/metabolights/MTBLS10551. Orthogonal partial least-square discriminant analysis (OPLS-DA) was used to build the model. The metabolic profile could be visualized as a score plot, where each point represents a sample. Differential metabolites were found from the list of primary substances in the sample and screened according to the preset P-value and variable importance projection (VIP) threshold in the statistical test (P < 0.05, VIP > 1). Pathway analysis of differential metabolites was performed by MetaboAnalyst (www.metaboanalyst.ca). The criterion for significant enrichment was P < 0.05.

16S rRNA Sequencing

According to the manufacturer's requirements, total bacterial genomic DNA was extracted from samples using the SteadyPure extraction kit (Accurate Biotechnology Co., Ltd, Changsha, China). The V3 and V4 regions of 16S rDNA gene were amplified by specific barcoded primers. The primer sequences were as follows: F:ACTCCTACGGGAGGCAGCA, R:GGACTACHVGGGTWTCTAAT. The PCR program was 94°C to 96°C for 1 min, followed by 35 cycles of 50°C to 60°C for 30 s, and final extension at 72°C for 1 min. The amplified products were purified by the AxyPrep DNA Gel Extraction Kit (Axygen, AZ), and quantified using LineGene 9600 Plus RealTime PCR System (Bioer Technology, Hangzhou, China). Then, 16S rRNA high-throughput sequencing was performed on Illumina Novaseq platform (Shanghai Personal Biotechnology Co., Ltd., Shanghai, China).

The DADA2 method was used for primer removal, quality filtering, denoising, splicing and chimera removal, and then sequencing data were compared with the Greengene database. Further analysis based on previous reports (Liu et al., 2023b). The R package was used to calculate the clustering results of each sample and classification unit, which is presented in the form of an interaction graph. The accession numbers for 16S rRNA is PRJNA1130356. Alpha diversity was analyzed using QIIME2 (https://github.com/QIIME2) software. Predicted functions were performed using the Encyclopedia of Genes and Genomes (KEGG) database (http://www.genome.jp/kegg/pathway.html), and the criterion for significant enrichment was P < 0.05.

Statistical Analysis

All analysis were performed with unpaired Student's t test using IBM SPSS Statistics 26.0 statistical software and plotted with GraphPad Prism 8. All data were expressed as mean ± standard error of mean (SEM), and P < 0.05 was considered statistically significant.

RESULTS

Growth Performance

As shown in Figure 2, DXM injection markedly decreased ADFI, laying rate, shell strength, albumen height, haugh unit, egg weight, folk weight and albumen weight (P < 0.05). Conversely, dietary betaine supplementation significantly increased laying rate, albumen height, haugh unit, egg weight, folk weight and albumen weight (P < 0.05).Figure 2 Growth performance for body weight, ADFI, laying rate, shell strength, albumen height, haugh unit, egg weight, folk weight and albumen weight. Data represent mean ± SEM (n = 10). *P < 0.05, **P < 0.01 versus Con; #P < 0.05, ##P < 0.01 vs. DXM.

Figure 2

Serum Biochemistry Indexes

The results about serum biochemistry indexes are shown in Figure 3. Following the injection of DXM, there was an upward trend observed in LDH, TBIL, TG, and LDL-c levels in the hens (P < 0.05 or 0.05 < P < 0.1). Whereas the levels of AST, LDH, TBIL and TBA were decreased in the BT group (P < 0.05 or 0.05 < P < 0.1).Figure 3 Serum biochemistry. (A–I) Serum biochemical parameters for AST, LDH, TBIL, TP, TBA, TC, TG, LDL-c, HDL-c. Data represent mean ± SEM (n = 10). Data represent mean ± SEM (n = 10). *P < 0.05, **P < 0.01 vs. Con; #P < 0.05, ##P < 0.01 vs. DXM.

Figure 3

Antioxidant Functions

DXM injection significantly reduced T-AOC and increased MDA levels in serum and liver when compared to the Con group (P < 0.05 or 0.05<P < 0.1). However, dietary betaine addition effectively restored these levels (P < 0.01 or P < 0.05) (Figure 4).Figure 4 Body antioxidant Indicators. (A-B) Serum antioxidant parameters for T-AOC and MDA. (C–D) Hepatic antioxidant parameters for T-AOC and MDA. Data represent mean ± SEM (n = 10). Data represent mean ± SEM (n = 10). *P < 0.05, **P < 0.01 vs. Con; #P < 0.05, ##P < 0.01 vs. DXM.

Figure 4

Hepatic Transcriptional Changes

To reveal hepatic transcriptional changes, RNA-seq was applied to identify DEGs and varied metabolic pathways. The number of up- and down- regulated genes were 88 and 110 respectively between Con and DXM groups, while 286 DEGs were identified between the DXM and BT groups, including 169 upregulation and 117 downregulation (Figure 5A and B). DEGs clustering analysis was performed and shown via the heat map where some genes disturbed by DXM injection were rescued by the addition of betaine, (Figure 5C and Supplementary Figure S1). The up-regulated enrichment pathways based on GSEA analysis were mainly cell adhesion molecules, glycerolipid metabolism and FOXO signaling pathway between Con and DXM groups (Figure 5D), and those pathways were down-regulated in BT group when compared with DXM (Figure 5E).Figure 5 DEGs identification and GSEA pathway enrichment analysis by transcriptomics. (A–B) Volcano plot of DEG among Con, DXM and BT groups. (C) The heatmap of DEGs. Up-regulated genes were shown in red, and blue represents down-regulated genes. (D–E) GSEA pathway enrichment from Con vs. DXM and DXM vs BT, respectively.

Figure 5

Differential Metabolites and KEGG Pathway Analysis

The OPLS-DA score plot showed that there was a clear separation among Con, DXM and BT groups (Figure 6A). A total of 44 and 94 differential metabolites were identified respectively from Con vs DXM and DXM vs BT (Figure 6B). Cluster analysis of differential metabolites heatmap indicated that some metabolites were changed and rescued by DXM or BT administration (Figures 6C and 6D). Some metabolites such as pantothenic acid, gamma-Aminobutyric acid, equol and choline were significantly increased in BT group which were associated with antioxidant and anti-inflammatory (Supplementary Table S1). Metabolic pathway enrichment analysis showed that pantothenate and CoA biosynthesis, glycerophospholipid metabolism, citrate cycle (TCA cycle), steroid hormone biosynthesis and glutathione metabolism were enriched in Con vs DXM (Figure 6E). while beta-alanine metabolism, pantothenate and CoA biosynthesis, glutathione metabolism and primary bile acid biosynthesis were enriched between DXM and BT groups (Figure 6F).Figure 6 Differential metabolites and pathway analysis based on serum metabolomics. (A) OPLS-DA score map of serum metabolites. (B) The number of differential metabolites among Con, DXM and BT groups. (C–D) Heatmap analysis of differential metabolites, presented as Con vs. DXM group and DXM vs. BT group, respectively. (E–F) Pathway enrichment based on differential metabolites from Con vs. DXM and DXM vs BT, respectively.

Figure 6

Cecum Microbiota Analysis

As shown in Figures 7A-C, the alpha-diversity indices including Chao 1, Faith_pd and Observed_species were remarkably higher in BT group when compared with the DXM group (P < 0.05). Heatmap analysis was further performed to compare the differences in species composition among 3 groups (Figure 7D). Results showed that the abundance of Subdoligranulum, Prevotella, Blautia, YRC22, Bacteroides, Ruminococcus and Coprococcus were significantly decreased in DXM group (P < 0.01 or 0.05 < P < 0.1), but dietary betaine addition significantly increased the abundance of these bacteria (P<0.05). Functional prediction analysis on level 3 indicated that tryptophan metabolism, valine, leucine and isoleucine degradation, lysine degradation and propanoate metabolism were enriched significantly in DXM group. Cyanamic acid metabolism was enhanced in BT group when compared with DXM group (Figures 7E and 7F).Figure 7 Gut microbiota composition analysis. (A–C) α-diversity of cecal microbiota by Chao 1, Faith_pd and Observed_species. (D) Species composition heat map among 3 groups. (E-F) Functional prediction analysis on level 3 of the metabolism pathway from Con vs. DXM and DXM vs. BT. Data represent mean ± SEM (n = 10). *P < 0.05, **P < 0.01 vs. Con; #P < 0.05, ##P < 0.01 vs. DXM.

Figure 7

DISCUSSION

Laying hens are usually exposed to a range of stressors due to environmental conditions and the laying physiology characteristics (Bai et al., 2023). A growing study clearly indicates that oxidative stress is a major detrimental consequence of the most common commercial stressors in poultry production (Surai et al., 2019) and ultimately impairs the growth performance (Min et al., 2016). DXM is an analogue of glucocorticoid that animals secrete in response to stress. It was reported that DXM can induce lipid metabolism disorders and aggravate oxidative stress (Lv et al., 2018), which was evidenced by elevated MDA and reduced T-AOC through the reduction of antioxidant enzymes activity and the increase in lipid peroxidation (Feng and Tang, 2014; Hasona and Morsi, 2019). In this study, lipid accumulation occurred after DXM injection, as indicated by increased serum TG and LDL-c. Simultaneously, serum T-AOC decreased, while LDH, TBIL and MDA contents increased, suggesting the occurrence of oxidative stress. However, these levels in BT group were notably reversed when compared with the DXM group. Moreover, the metabolomics results revealed changes in serum metabolites, including chavicol (Santos et al., 2018), salicylic acid (Zhang et al., 2023), 2-pyrocatechuic acid (Nenadis et al., 2022), citric acid (Wu et al., 2018), luteolin 7-O-beta-D-glucoside (Moniruzzaman et al., 2018), and ergothioneine (Halliwell et al., 2018), all of which are associated with antioxidant and anti-inflammatory. These metabolites exhibited a decrease following DXM injection. Although these metabolites were not directly reversed by dietary betaine addition, pantothenic acid (Tutun et al., 2019), gamma-Aminobutyric acid (Fathi et al., 2023), equol (Gou et al., 2015) and choline (Dong et al., 2019) related to antioxidant and anti-inflammatory were upregulated. These results demonstrate that DXM can induce oxidative stress in laying hens, while betaine can alleviate oxidative stress and liver damage by reducing lipid peroxidation products and increasing the contents of metabolites associated with antioxidants. Wen et al. (2021) found that dietary betaine supplementation can alleviates heat stress-induced hepatic damaged by regulating antioxidant enzyme system. Balkan et al. (2004) reported that betaine restored the changes in triglyceride, lipid peroxide and GSH levels following ethanol ingestion treated guinea pigs. These findings are consistent with our studies.

It has been reported that high levels of glucocorticoids reduce egg production (El-Lethey et al., 2003). DXM, as a synthetic glucocorticoid, can negatively affect eggs by inhibiting ovulation and suppressing the production of luteinizing hormone and progesterone. Additionally, oxidative stress has been reported to adversely effect on egg quality traits (Jing et al., 2022) and causes decline in the internal and external quality characteristics of the egg by reducing the availability of essential substances, such as water, proteins, lipids, carbohydrates, vitamins and minerals (Eid et al., 2021). In agreement with our study, the inclusion of DXM injection resulted in a lower laying rate, shell strength, albumen height, haugh unit, egg weight, folk weight and albumen weight, which could be partly rescued by dietary betaine addition (Ratriyanto and Mosenthin, 2018; Guo et al., 2023).

Hepatic triglycerides (TG) synthesis is necessary for yolk formation by packaging into very-low density lipoprotein (VLDL) and transportation into ovarian ducts (Dong et al., 2019). During egg laying period, the liver of laying hens tolerates a large amount of lipid and undergoes enhanced lipid metabolic activity, which will be susceptible to oxidative stress (Yonke and Cherian, 2019). In the present study, decreased T-AOC and increased MDA levels in the liver and serum indicated the imbalance of the oxidative stress system of birds. Transcriptomic results showed that cell adhesion molecules, glycerolipid metabolism, FOXO signaling pathway and other pathways were significantly upregulated after DXM injection. However, dietary betaine addition appeared to ameliorate hepatic oxidative status by reversing the expression of genes associated with lipid metabolism (ACVR1C, UGGT2, C14orf180) (Kerr et al., 2019; Tian et al., 2021; Hung et al., 2022) and down-regulating pathways such as cell adhesion molecules, glycerolipid metabolism and glycolytic gluconeogenesis. The response to betaine observed in this study may be attributed to its function as a methyl donor to homocysteine, thereby facilitating glutathione production, which subsequently stabilizes lipid peroxidation.

As the organ with the largest surface area exposed to the external environment, the intestine harbors lots of microorganisms and functions as the principal site for diet digestion, nutrients absorption and metabolism, rendering it particularly vulnerable to oxidative stress induced by oxygen free radicals. This susceptibility underscores the potential for significant long-term damage to the intestine from the consequences of oxidative stress. Previous studies have found that dietary betaine addition can reduce oxidative damage caused by heat stress through enhancing the colonization of beneficial bacteria (Al Sulaiman et al., 2024). Likewise, the current research manifested that dietary betaine addition significantly augmented the abundance of beneficial bacteria such as Subdoligranulum, Blautia, Bacteroides, Coprococcus, YRC22 in hens under the condition of DXM injection. It has been reported that Subdoligranulum is a butyric acid-producing bacterium (Singh et al., 2023), and butyric acid and has an important role in gastrointestinal health (Hodgkinson et al., 2023). Blautia has been reported inversely correlated with obeity (Hosomi et al., 2022). Liu et al. found (Liu et al., 2021) that Blautia is a new functional genus with potential probiotic properties. Previous research found that Coprococcus is a potent probiotic which can alleviates colitis (Yang et al., 2023). Recent study highlights that the abundance of YRC22 were down-regulated in the fatty liver group (Liu et al., 2023a). These results indicated that betaine can ameliorate stress-induced dysbiosis via remodeling gut microbiota composition, which might be attributed to providing additional carbon sources for the growth and proliferation of symbiotic bacteria in the intestines.

CONCLUSIONS

In summary, our findings demonstrate that DXM treatment indeed causes oxidative stress and lipid metabolism disorder, which results in the loss of egg production performance and quality. However, dietary betaine supplementation ameliorated various parameters related to egg quality via regulating hepatic metabolism changes and remodeling gut microbiota composition. The mechanism might involve in the effect of betaine on oxidative stress-related genes expression and metabolites. Overall, betaine shows promise as a dietary intervention to counteract oxidative stress and improve egg laying performance in laying hens.

DISCLOSURES

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Appendix Supplementary materials

Image, application 1

Image, application 2

ACKNOWLEDGMENTS

This work was funded by the National Science Foundation of China (32372910 and 32102567 ); the Program for Shaanxi Science & Technology (2023-YBNY-144 , 2022GD-TSLD-46-0302 , K3031223077 and 2023BSHEDZZ151 ); China Postdoctoral Science Foundation (2023M742862 ).

Supplementary material associated with this article can be found in the online version at doi:10.1016/j.psj.2024.104178.
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