The poultry ovary is used as a classic model to study ovarian biology and ovarian cancer. Stress factors induced oxidative stress to cause follicle atresia, which may be a fundamental reason for the reduction in fertility in older laying hens or in aging women. In the present study, we set out to characterize the relationships between oxidative stress and ovarian function. Layers (62 weeks of age; BW = 1.42 ± 0.12 kg) were injected with tert-butyl hydroperoxide (tBHP) at 0 (CON) and 800 μmol/kg BW (oxidative stress group, OS) for 24 days and the role of melatonin (Mel) on tBHP-induced ovary oxidative stress was assessed through ovary culture in vitro. The OS (800 μmol/kg BW tert-butyl hydroperoxide) treatment decreased the reproduction performance and ovarian follicle numbers. OS decreased the expression of SIRT1 and increased the P53 and FoxO1 expression of the ovary. A decreased Firmicutes to Bacteroidetes ratio, enriched Marinifilaceae (family), Odoribacter (genus) and Bacteroides_plebeius (species) were observed in the cecum of the OS group. Using Mel in vitro enhanced the follicle numbers and decreased the ovary cell apoptosis induced by tBHP. In addition, it increased the expression of SIRT1 and decreased the P53 and FoxO1 expression. These findings indicated that oxidative stress could decrease the laying performance, ovarian function and influence gut microbiota and body metabolites in the layer model, while the melatonin exerts an amelioration the ovary oxidative stress through SIRT1-P53/FoxO1 pathway.

A total of 360-day-old broiler chicks were allocated into six groups in 2 (Coccidial challenge or not) × 3 (dietary treatments) factorial design. Three dietary treatments including: basic diet, basic diet plus organic acids (OAs) in drinking water, and basic diet plus OAs in the feed with and without coccidial challenge. The OAs in water or feed improved (P < 0.01) average body weight (ABW), average body weight gain (ABWG), and feed conversion ratio (FCR) as compared with the control diet during starter, grower, and whole experimental period. Coccidial challenge decreased BW, ABWG, and average feed intake (AFI), as well as resulted in poor FCR during the starter and whole experimental period (P < 0.05). Though there was no interaction between OAs supplementation and coccidial challenge, the OAs supplementation improved the overall performance with and without coccidial challenge birds on 21 d and 35 d. IgG was found higher (P = 0.03) in broilers fed OAs in feed without the coccidial challenge group. On 18 d, OAs supplementation in feed increased TNF-γ (P = 0.006), whereas the coccidial challenge decreases TNF-γ (P = 0.01) and IL-10 (P = < .0001), and increases IgM (P = 0.03), IgG (P = 0.04) and IgA (P = 0.02). On 29 d, the coccidial challenge increases IgM and IgA. On 18 d, jejunal lesion score was found significantly higher in the coccidial challenged group as compared to OAs supplementation with coccidial challenged groups on 18 d (P < 0.0001) and 29 d (P = 0.03). Crypt depth was higher, and Villus-height to Crypt depth ratio was lower in the coccidial challenge group on 18 and 29 d. The Goblet cells were found higher in the non-coccidial challenge on 18 d. After 18 d, 16S rDNA gene sequence analysis of ileal chyme has shown that coccidial challenge decreases Lactobacillus_reuteri species as compared to the non-challenged group (P = 0.02). After 29, Cyanobacteria abundance reduced (P = 0.014) in the challenged group than the non-challenged group at the phylum level. At the genus level, Lactobacillus (P = 0.036) and unidentified Cyanobacteria (P = 0.01) were found higher in the non-challenged group than the coccidial challenge group. The results indicate that the OAs supplementation showed improved responses in a pattern similar to the non-challenged control group by neutralizing the negative effects of the coccidial challenge.

Resveratrol (RV) is associated with protection against oxidative stress to improve health, however the effect of RV in layers under oxidative stress (OS) is limited. The objective of this experiment was to investigate the negative effect of OS and protective effects of RV against OS in laying hens. 40 Lohmann layers (25-wk-old; BW = 1.44±0.10 kg) were allocated to four treatments in a 2 × 2 factorial arrangement with either RV (0 or 600 mg/kg) or intraperitoneal injection of tert-butyl hydroperoxide (tBHP) (0 or 800 μmol/kg BW) for 31 days. The results shown that the hens challenged with tBHP presented lower egg-laying rate, feed intake, feed efficiency and higher defective egg rate (P_(tBHP)<0.05). The RV were also observed to attenuated egg laying rate and feed intake reduction together with decreased broken egg rate under t-BHP challenge (P_{(Interaction)}≤0.01). The tBHP challenged layer demonstrated lower intestinal morphology (villus height in duodenum and jejunum), lower antioxidant enzymes activities [total superoxidase (SOD), glutathione peroxidase (GSH-Px), total antioxidant capacity (T-AOC)], and glutathione (GSH) levels and higher malondialdehyde (MDA) level] (P_(tBHP)<0.05). Dietary RV increased jejunal SOD, GSH-Px and T-AOC activities, and reduced MDA concentration (P_(RV) ≤0.05). Layers under tBHP challenge up-regulated mRNA expression of pro-inflammatory cytokine [interleukin-1β (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α)] and nuclear factor NF-κB (P_(tBHP)<0.05) in jejunum. Dietary RV supplementation down-regulated mRNA gene expression of IL-1β, IL-6, TNF-α and NF-κB (P_(RV) ≤0.05). Dietary RV up-regulated mRNA expression of jejunal barrier-related proteins (claudin-1, claudin-2, mucin-1, and occludin) and ovarian reproductive hormone receptor [steroidogenic acute regulatory protein (StAR), androgen receptor (AR), estrogen receptor 1 (ESR1), and activin a receptor type 1 (ACVR1)] (P_(RV) ≤0.05). Overall, the results indicate that tBHP induced oxidative stress to result in reducing production performance, intestinal health and induced ovarian inflammation; whereas dietary RV was able to maintain intestinal health and mitigate the negative impact of tBHP challenge on production performance and ovarian function.

This study was conducted to determine the effects of supplemental dietary benzoic acid on production performance, egg quality, intestinal morphology, and intestinal microbiota of laying hens. A total of seven hundred twenty 45-wk-old Lohman pink-shell laying hens were randomly allocated to 3 dietary treatments: control (CON), diet supplemented with 1,000 mg/kg benzoic acid (BA1), and 2,000 mg/kg benzoic acid (BA2). Each treatment included 10 replicates of 24 hens; laying hens were monitored for 16 wk. Overall, the results indicate that benzoic acid supplementation had no effect on laying rate, feed intake, feed conversion ratio, and breaking rate; however, a decrease in egg weight (P < 0.01) was observed in the BA2 group. Albumen height and Haugh unit (HU) were also linearly increased in the BA1 and BA2 groups (linear effect, P < 0.05). An increase in duodenum villus height (V) (quadratic effect, P = 0.041) and crypt depth (C) (linear effect, P = 0.012) was observed in the BA2 group, whereas an increased jejunum C and decreased V/C (quadratic effect, P < 0.05) in the BA1 group. Moreover, an increase in ileum V and C (quadratic effect, P < 0.05) was observed in the BA1 group. Microbial richness and diversity were reduced in the BA2 group (P < 0.01). An increase in the abundance of Clostridia (class), Clostridiales (order), Ruminococcaceae (family), and Lachnospiraceae (family) was noted in the BA1 group, whereas an enrichment of Bacteroides caecicola (species) was observed in the BA2 group. The HU positively correlated with genus Sphaerochaeta and Enorma (r = 0.56, 0.56; P < 0.05) but negatively correlated with Romboutsia, Subdoligranulum, Helicobacter, and Mucispirillum (r = −0.58, −0.49, −0.48; −0.70; P < 0.05). In conclusion, dietary supplementation with benzoic acid had no effect on production performance, but it significantly improved egg quality. In addition, 1,000 mg/kg benzoic acid positively modulated intestinal health by improving intestinal morphology and enriching microbial composition.

Zearalenone (ZEA) is a nonsteroidal estrogenic mycotoxin found in several food commodities worldwide. ZEA causes reproductive disorders, genotoxicity, and testicular toxicity in animals. However, little is known about the functions of apoptosis and autophagy after exposure to ZEA in granulosa cells. This study investigated the effects of ZEA on chicken granulosa cells. The results show that ZEA at different doses significantly inhibited the growth of chicken granulosa cells by inducing apoptosis. ZEA treatment up-regulated Bax and downregulated Bcl-2 expression, promoted cytochrome c release into the cytosol, and triggered mitochondria-mediated apoptosis. Consequently, caspase-9 and downstream effector caspase-3 were activated, resulting in chicken granulosa cells apoptosis. ZEA treatment also upregulated LC3-II and Beclin-1 expression, suggesting that ZEA induced a high level of autophagy. Pretreatment with chloroquine (an autophagy inhibitor) and rapamycin (an autophagy inducer) increased and decreased the rate of apoptosis, respectively, in contrast with other ZEA-treated groups. Autophagy delayed apoptosis in the ZEA-treated cells. Therefore, autophagy may prevent cells from undergoing apoptosis by reducing ZEA-induced cytotoxicity. In addition, our results further show that the autophagy was stimulated by ZEA through PI3K-AKT-mTOR and MAPK signaling pathways in chicken granulosa cells.

This study was conducted to determine the effects of 25-hydroxycholecalciferol (25-OH-D3) on performance, egg quality, tibia quality, and serum hormones concentration in laying hens reared under high stocking density. A total of 800 45-week-old Lohmann laying hens were randomly allotted into a 2 × 2 factorial design with 2 levels of dietary 25-OH-D3 levels (0 and 69 μg/kg) and 2 rates of stocking densities [506 (low density) and 338 (high density) cm2/hen]. Laying hens were monitored for 16 wk. High stocking density decreased laying rate, egg weight, and feed intake compared with low stocking density (P < 0.01) during 1 to 8 wk and 1 to 16 wk. Overall, high stocking density increased eggshell lightness value and decreased shell redness and yellowness value, strength, thickness, and relative weight compared with low stocking density (P < 0.05). Dietary supplementation with 25-OH-D3 reduced the value of the eggshell lightness and increased its yellowness and eggshells weight (P ≤ 0.05). The increase in eggshell thickness was more pronounced when 25-OH-D3 was supplemented to layers under high stocking density (interaction, P < 0.05). Layers under high stocking density had lower ash content and calcium content in the tibia than layers under low stocking density (P = 0.04); dietary 25-OH-D3 increased tibia strength compared with no addition (P = 0.05). Layers under high stocking density had higher serum concentrations of 25-OH-D3, corticosterone (CORT), lipopolysaccharide (LPS), and osteocalcin (OC; P < 0.05), lower content of parathyroid hormone (PTH) compared with layers under low stocking density (P < 0.01). Dietary 25-OH-D3 increased serum concentration of 25-OH-D3, carbonic anhydrase (CA), and calcitonin (CT) (P < 0.01) and reduced corticosterone, lipopolysaccharide and osteocalcin concentration (P ≤ 0.05). The increase effect in PTH was more pronounced when 25-OH-D3 was supplemented to layers under high stocking density (interaction, P = 0.05). Overall, the results gathered in this study indicate that high stocking density result in reducing production performance, shell color and quality, and tibia health, whereas dietary 25-OH-D3 was able to maintain tibia health and to mitigate the negative impact of high stocking density on productive performance.