Haidong Du

Immune stress under intensive high density production conditions impairs broiler health and performance, highlighting the need for effective nutritional strategies. This study established an immune stress model by intraperitoneal lipopolysaccharide (LPS) injection to investigate the mechanism by which Glycyrrhiza polysaccharides (GPs) alleviate LPS-induced immune stress. A total of 240 AA broilers were randomly assigned to four groups: control (CON), GP, LPS, and GP + LPS, with six replicates per group and ten birds per replicate. During the stress phase (28 d), LPS challenge increased serum ACTH and CORT concentrations and altered several immune-related parameters; whereas, GP supplementation improved ileal morphology and reduced serum IL-1β, NF-κB, and TNF-α concentrations. Significant GP × LPS interactions were observed for IL-4, IgA, IgG, and iNOS. During the recovery phase (35 d), GP supplementation increased serum IgA and decreased iNOS, while significant GP × LPS interactions were detected for IL-1β, NF-κB, and sCD4. Notably, GP attenuated the LPS-associated increase in NF-κB. Integrated multi-omics analysis indicated coordinated regulation at transcriptional and proteomic levels. GP upregulated FOS, JUN, and CATH3 and enriched NF-κB and Wnt signaling at the transcript level, while increasing IRF1 and CATH3 and decreasing CTNNBIP1 at the protein level, primarily involving Wnt and MAPK pathways. Joint analysis identified CATH3 as a central regulatory target. Collectively, GP alleviates immune stress by restraining excessive inflammation and modulating Wnt/MAPK signaling, thereby promoting intestinal structural recovery.

This study investigated changes in physiological processes and rumen microbial communities in does from mid-gestation to lactation and identified potential associations between these physiological changes and the rumen microbiome. Additionally, we studied the transmission mechanisms of microorganisms between the dam and offspring. Our study demonstrates significant changes in maternal physiological metabolism, immune status, and rumen microbiota from mid-pregnancy through lactation. We identified potential associations between these physiological changes and the rumen microbiome. Moreover, the findings highlight that alterations in maternal physiological metabolism and immune status significantly influence the growth and immune development of offspring kids. Additionally, we observed that the maternal microbiota serves as a key source of gastrointestinal microbial communities in young animals, with early colonization of maternally derived microbes in the offspring’s gastrointestinal tract playing a role in shaping their immune system development. The results for primary outcomes are as follows: The serum levels of estrogen and progesterone in pregnant does were greater than those observed during lactation, while the concentration of growth hormone, triiodothyronine, and glucose exhibited an upward trend during lactation. During late gestation, the serum IL-10 concentration in does decreased, while the TNF-α concentration increased. Additionally, on day 140 of gestation, does showed a significant decrease in IgG, total protein, and globulin levels. From mid-gestation to lactation, the abundance of dominant phyla and genera, including Firmicutes, Bacteroidetes, Patescibacteria, Bacteroidales_RF16_group, Clostridia_UCG-014, RF39, and Eubacterium_ventriosum_group, in the rumen of does underwent significant changes. LEfSe analysis identified a series of marker microorganisms in the rumen of does at different physiological stages. A correlation was observed between these dominant bacteria and the serum physiological indicators of the does. Notably, rumen volatile fatty acids also exhibited a correlation with serum physiological indicators. In addition, serum physiological indicators of does were significantly correlated with the growth and immune indicators of their kids. Microbiological origin analysis revealed that the gastrointestinal microbiome of kids primarily originated from the rumen, birth canal, and milk of does. Further analysis identified a correlation between the kids’ serum immunometric indicators and certain gastrointestinal microorganisms. In particular, the jejunum microbiota of 28-day-old lactating kids, including Alysiella, Neisseria, and Muribaculaceae, showed a significant positive correlation with serum IL-6 and IL-10 levels. Meanwhile, these genera were dominant in the saliva and milk of does, suggesting a direct microbial transfer from dam to offspring. These microbial communities may play a significant role in modulating the metabolism and immune responses of the offspring, thereby influencing their immune system development.

Maternal metabolic intensity significantly increases during late gestation and lactation, placing significant stress on cells and tissues. This heightened metabolic demand can lead to inflammatory responses and metabolic disorders, adversely affecting the health of both the mother and her offspring. Diet plays a key role in modulating host health by influencing the gastrointestinal microbiome. This study examined the impact of two roughage sources, corn straw (CS), and alfalfa hay (AH), on ewes and their offspring during late gestation and lactation, with a focus on metabolism, immunity, and the microbiome. Thirty-six multiparous Inner Mongolia cashmere goats, approximately 60 days pregnant, were assigned to CS and AH groups. Samples were collected from the ewes on day 140 of gestation (G140) and day 28 of lactation (L28) for analysis. The results showed that ewes fed AH had reduced body weight loss during lactation (p < 0.05), and increased serum metabolic factors levels (p < 0.05). Additionally, ewes in the AH group exhibited a reduced inflammatory response during both gestation and lactation compared to those in the CS group, as evidenced by a significant decrease in TNF-α and LPS levels and a notable increase in IL-10 (p < 0.05). The rumen microbiomes of ewes in the AH and CS groups exhibited stark differences, with specific microbial markers identified at G140 and L28. Correlation analysis revealed associations between microbiome, volatile fatty acids, cytokines, and metabolic markers. The analysis of the lambs demonstrated that their immune status and microbial composition were significantly influenced by the immune health and microbial community structure of the ewe. Moreover, microbial and immune-related components from the ewes were transmitted to the lambs, further shaping their immune development and rumen microbiota. Overall, different roughage sources during late gestation and lactation had minimal impact on the growth performance of ewes and lambs, given that both diets were iso-nitrogen and iso-energetic. However, ewes fed AH exhibited significant improvements in immune function and overall health for both them and their lambs.

During pregnancy and lactation, maternal nutrition is linked to the full development of offspring and may have long-term or lifelong effects. However, the influence of the doe’s diet on the gastrointestinal (GI) tract of young kids remains largely unexplored. Therefore, we investigated the effects of doe roughage sources (alfalfa hay, AH, or corn straw, CS) during pregnancy and lactation on kid growth, GI morphology, barrier function, metabolism, immunity, and microbiome composition. The results indicate that, compared with the CS group, does fed an AH diet had significantly higher feed intake (p < 0.01). However, CS-fed does exhibited higher neutral detergent fiber (NDF) digestibility (p < 0.05). There were no significant differences in animal (doe or kid) weight among the groups (p > 0.05). In the rumen of goat kids, the AH group exhibited a higher papillae width and increased levels of interleukin-10 (IL-10) compared with the CS group (p < 0.05). In the jejunum of goat kids, the AH group showed a higher villus-height-to-crypt-depth (VH/CD) ratio, as well as elevated levels of secretory immunoglobulin A (SIgA), immunoglobulin G (IgG), IL-10, acetate, and total volatile fatty acids (TVFAs), when compared with the CS group (p < 0.05). Transcriptome analysis revealed that the source of roughage in does was associated with changes in the GI transcriptome of the kids. Differentially expressed genes (DEGs) in the rumen were mainly associated with tissue development and immune regulation, while the DEGs in the jejunum were mainly associated with the regulation of transferase activity. Spearman correlation analyses indicated significant associations between GI DEGs and phenotypic indicators related to GI development, immunity, and metabolism. LEfSe analysis identified 14 rumen microbial biomarkers and 6 jejunum microbial biomarkers. Notably, these microorganisms were also enriched in the rumen or day 28 milk of the does. Further microbial composition analysis revealed significant correlations between the rumen and milk microbiomes of does and the rumen or jejunum microbiomes of kids. Association analyses indicated that microbial biomarkers interact with host genes, thereby affecting the development and function of the GI system. Additionally, correlation analyses revealed significant association between milk metabolites and the rumen and jejunum microbiomes of kids. This study demonstrated that maternal diet significantly influences the development of microbial ecosystems in offspring by modulating microbial communities and metabolite composition. The early colonization of GI microorganisms is crucial for the structural development, barrier function, immune capacity, and microbial metabolic activity of the GI system.

Alfalfa silage due to its high protein can lead to easier feeding management, but its high proportion of rumen-degradable protein can reduce rumen nitrogen utilization. Nevertheless, increasing dietary energy can enhance ruminal microbial protein synthesis. Thirty-two Suffolk female sheep were used in this study, with a 2 × 2 factorial arrangement of treatment. The four treatments were a combination of two forage types (alfalfa hay; AH vs. alfalfa silage; AS) and two rumen-degradable starch levels (low RDS; LR vs. high RDS; HR) with a 15 d adaptation and 60 d experimental period. The rumen content and rumen epithelium samples were collected after slaughter. Feeding AS increased the rumen isobutyrate, valerate, ammonia-N (NH₃-N) concentration, urase activity, and papillae height (p < 0.05) and reduced the feed to gain (F:G), rumen bacterial protein (BCP), rumen lactic acid concentration, and papillae width (p < 0.05) of sheep. Increased RDS in the diet improved the daily matter intake, average daily gain, and rumen weight, reduced the F:G, and enhanced the rumen nitrogen capture rate by decreasing total amino acids and the NH₃-N concentration to increase BCP, aquaporins 3 gene, and protein expression. The rumen microbiota also changed as the HR diet reduced the Chao index (p < 0.05). The metabolomics analysis showed that feeding AS upregulated the rumen tryptophan metabolism and steroid hormone biosynthesis, while the purine metabolism, linoleic acid metabolism, and amino acid biosynthesis were downregulated. Furthermore, increased RDS in the diet upregulated rumen lysine degradation and sphingolipid metabolism, while aromatic amino acid biosynthesis was downregulated. Additionally, the correlation analysis results showed that ADG was positively correlated with 5-aminopentanoic acid, and three microorganisms (unclassified_f__Selenomonadaceae, Quinella, Christensenellaceae_R-7_group) were positively correlated with the rumen isobutyrate, valerate, NH₃-N concentration, urase activity, tryptophan metabolism, and steroid hormone biosynthesis and negatively correlated with linoleic acid metabolism and amino acid biosynthesis in sheep. In summary, increased RDS in the diet improved the growth performance and rumen N utilization and reduced bacterial diversity in sheep. The alfalfa silage diet only increased feed efficiency; it did not affect growth performance. Additionally, it decreased rumen nitrogen utilization, linoleic acid, and amino acid biosynthesis. Nevertheless, there were limited interactions between forage and RDS; increased RDS in the AS diet enhanced the nitrogen capture rate of rumen microorganisms for alfalfa silage, with only slight improvements in the purine metabolism, linoleic acid, and amino acid synthesis.

The spleen and small intestines are the primary immune organs that provide important immunity against various diseases. Artemisia ordosica polysaccharide (AOP) could be used as an immunologic enhancer to boost immunity in response to infection. This study was performed to explore the effects of the dietary supplementation of AOP on the growth performance and spleen and small intestine immune function in broilers. A total of 288 AA broilers (1 day old) were randomly assigned into six dietary groups. Each group included six replicates of eight broilers per cage. The broilers were fed with a basal diet supplemented with 0 mg/kg (CON), 50 mg/kg chlortetracycline (CTC), 250, 500, 750, and 1000 mg/kg AOP for 42 d. The results showed that dietary AOP supplementation affected broiler growth performance, with 750 and 1000 mg/kg of AOP being able to significantly improve broiler BWG, and 750 mg/kg of AOP was able to significantly reduce the FCR. The dietary AOP supplementation increased the levels of IgA, IgG, IgM, IL-1β, IL-2, and IL-4 in the spleen and small intestine in a dose-dependent manner (p < 0.05). Meanwhile, we found that AOP can promote the mRNA expression of TLR4/MAPK/NF-κB signaling-pathway-related factors (TLR4, MyD88, P38 MAPK, JNK, NF-κB p50, and IL-1β). In addition, the dietary supplementation of 750 mg/kg AOP provides better immunity in the tissue than the CON group but showed no significant difference from the CTC group. Therefore, AOP has an immunoregulatory action and can modulate the immune function of broilers via the TLR4/ NF-ΚB/MAPK signal pathway. In conclusion, dietary supplementation with 750 mg/kg AOP may be alternatives to antibiotics for enhancing broilers’ health, immunity, and growth performance.