Copper (Cu) serves as an essential cofactor in all organisms, yet excessive Cu exposure is widely recognized for its role in inducing liver inflammation. However, the precise mechanism by which Cu triggers liver inflammation in ducks, particularly in relation to the interplay in gut microbiota regulation, has remained elusive. In this investigation, we sought to elucidate the impact of Cu exposure on liver inflammation through gut-liver axis in ducks. Our findings revealed that Cu exposure markedly elevated liver AST and ALT levels and induced liver inflammation through upregulating pro-inflammatory cytokines (IL-1β, IL-6 and TNF-α) and triggering the LPS/TLR4/NF-κB signaling pathway. Simultaneously, Cu exposure induced alterations in the composition of intestinal flora communities, notably increasing the relative abundance of Sphingobacterium, Campylobacter, Acinetobacter and reducing the relative abundance of Lactobacillus. Cu exposure significantly decreased the protein expression related to intestinal barrier (Occludin, Claudin-1 and ZO-1) and promoted the secretion of intestinal pro-inflammatory cytokines. Furthermore, correlation analysis was observed that intestinal microbiome and gut barrier induced by Cu were closely related to liver inflammation. Fecal microbiota transplantation (FMT) experiments further demonstrated the microbiota-depleted ducks transplanting fecal samples from Cu-exposed ducks disturbed the intestinal dysfunction, which lead to impaire liver function and activate the liver inflammation. Our study provided insights into the mechanism by which Cu exposure induced liver inflammation in ducks through the regulation of gut-liver axis. These results enhanced our comprehension of the potential mechanisms driving Cu-induced hepatotoxicity in avian species.

Background: Cage-rearing in laying ducks, as a novel rearing system, not only fundamentally solves the pollution problem of the duck industry and improve bio-safety and product quality but also exhibits more benefits by implementing standardized production compared with the floor-rearing. Of course, this system also brings some welfare problems and stress injuries to layers due to lack of water environment and limited activities in the cages. However, the effects on the factors of antioxidant defense and inflammatory injury in the early cage stage are not well-understood. Results: In this study, eighty Shaoxing layers were reared on floor and in cages from 12 weeks of age. The ducks were caged 1, 2, 4, 7, and 10 days, the factors of antioxidant defense and inflammatory injury were investigated. The results showed that the caged ducks suffered liver injury to a certain extent when the ducks were just put into the cages. Analysis of antioxidant enzyme activities indicated that the different rearing system could not affect the change of antioxidant capacities, while the liver malondialdehyde (MDA) level was significant higher in the 2-d, 7-d, and 10-d ducks compared with the 1-d ducks during the change of days, while catalase (CAT) activity showed the opposite results. Additionally, quantitative real-time PCR (qRT-RCR) revealed that the relative mRNA levels of endoplasmic reticulum (ER) stress-related gene (CHOP and GRP78) were significantly upregulated in cage rearing ducks compared to that of the floor rearing ducks. Moreover, the mRNA levels of inflammatory cytokines including cycloxygenase-2 (COX-2), nitric oxide synthase (iNOS), Interleukin 1 beta (IL-1β), Interleukin 2 (IL-2) and Interleukin 6 (IL-6), were also increased significantly in caged layers. Conclusions: Taken together, although antioxidant defense has no obvious effect on cage stress, the stress levels of laying ducks vary greatly in the early cage stage, which not only caused liver tissue damage to some extent, but also resulted in increases in the expression of the factors of inflammatory injury. Therefore, we recommend that anti-stress agents should be added in the feed to alleviate the stress in the early cage stage.

Fulvic acid (FA) is a mixture of polyphenolic acid compounds extracted from humus, peat, lignite, and aquatic environments; it is used in traditional medicine to treat digestive tract diseases. The purpose of the present study was to investigate the effect of FA on growth performance, inflammation, intestinal microbiota, and metabolites in Xianju yellow chicken. The 240 Xianju yellow chickens (age, 524 days) included were randomly categorized into 4 treatments with 6 replicates per treatment and 10 birds per replicate. Birds received a basal diet or a diet supplemented with 500, 1,000, or 1,500 mg/kg of FA, for a period of 42 days. Dietary supplementation of FA improved average daily gain (ADG) and feed conversion ratio (FCR) (P> 0.05). Compared with the control group, the serum level of TNF-α in birds supplemented with FA was significantly decreased (P< 0.05), and that of IL-2 was significantly increased after administration of 1,500 mg/kg FA (P< 0.05). Analysis of gut microbiota indicated that FA reduced the relative abundance of genusMucispirillum, Anaerofustis, andCampylobacter, but enriched genusLachnoclostridium, Subdoligranulum, Sphaerochaeta, Oscillibacter, andCatenibacillusamong others. Untargeted metabolomic analyses revealed that FA increased 7-sulfocholic acid, but reduced the levels of Taurochenodeoxycholate-7-sulfate, LysoPC 20:4 (8Z, 11Z, 14Z, 17Z), LysoPC 18:2, Phosphocholine and other 13 metabolites in the cecum. The results demonstrated that FA may potentially have a significant positive effect on the growth performance and immune function of Xianju yellow chicken through the modulation of the gut microbiota.

Objective: Tibetan chickens, which have unique adaptations to extreme high-altitude environments, exhibit phenotypic and physiological characteristics that are distinct from those of lowland chickens. However, the mechanisms underlying hypoxic adaptation in the liver of chickens remain unknown.Methods: RNA-sequencing (RNA-Seq) technology was used to assess the differentially expressed genes (DEGs) involved in hypoxia adaptation in highland chickens (native Tibetan chicken [HT]) and lowland chickens (Langshan chicken [LS], Beijing You chicken [BJ], Qingyuan Partridge chicken [QY], and Chahua chicken [CH]).Results: A total of 352 co-DEGs were specifically screened between HT and four native lowland chicken breeds. Gene ontology and Kyoto encyclopedia of genes and genomes enrichment analyses indicated that these co-DEGs were widely involved in lipid metabolism processes, such as the peroxisome proliferator-activated receptors (PPAR) signaling pathway, fatty acid degradation, fatty acid metabolism and fatty acid biosynthesis. To further determine the relationship from the 352 co-DEGs, protein-protein interaction network was carried out and identified eight genes (ACSL1, CPT1A, ACOX1, PPARC1A, SCD, ACSBG2, ACACA, and FASN) as the potential regulating genes that are responsible for the altitude difference between the HT and other four lowland chicken breeds.Conclusion: This study provides novel insights into the molecular mechanisms regulating hypoxia adaptation via lipid metabolism in Tibetan chickens and other highland animals.

Objective: This study was designed to investigate the hypothesis that dietary quercetin (QUE) and coated sodium butyrate (SB) supplementation alleviate oxidative stress in the small intestine of diquat (DIQ)-challenged pullets.Methods: A total of 200 13-week-old pullets were divided into four groups: the control group (CON), the DIQ group, the QUE group, and the coated SB group, and injected intraperitoneally with either saline (CON) or diquat (DIQ, QUE, and SB) to induce oxidative stress on day 0.Results: On the first day, the malondialdehyde and superoxide dismutase (SOD) concentrations in the SB group were significantly different from those in the DIQ and QUE groups (p<0.05), and dietary supplementation with SB increased serum glutathione peroxidase (GSH-PX) levels compared with the DIQ group (p<0.05). Quercetin and SB increased the levels of CLAUDIN-1 and zonula occludens-1 (ZO-1) in the jejunum. On the tenth day of treatment, QUE attenuated the decrease in GSH-PX levels compared to those of the CON group (p<0.05), while SB increased SOD, GSH-PX, and total antioxidant capacity levels compared to those of the DIQ group. Nuclear factor erythroid 2-related factor 2 (NRF2) and heme oxygenase-1 (HO-1) mRNA levels in the QUE and SB groups increased (p<0.05) and CLAUDIN-1 mRNA levels in the QUE and SB groups were upregulated compared to those in the DIQ group ileum tissue.Conclusion: Supplementation of QUE and SB demonstrated the ability to relieve oxidative stress in pullets post DIQ-injection with a time-dependent manner and QUE and SB may be potential antioxidant additives for relieving oxidative stress and protecting the intestinal barrier of pullets.