Animals

Insulin-like growth factor 2 mRNA-binding protein 2 (IGF2BP2) is an RNA-binding protein known to play critical roles in metabolism, cell proliferation, and tumorigenesis. Although its involvement in muscle development has been documented in several species, the function of goose IGF2BP2 remains largely unexplored. In this study, we cloned and characterized the full-length cDNA and genomic DNA sequences of goose IGF2BP2. The cDNA is 2957 bp in length and contains a 1662 bp open reading frame encoding a 553-amino acid protein with five conserved RNA-binding domains. The genomic sequence spans 12,183 bp and consists of 12 exons and 11 introns. A total of 60 genetic variants were identified, including a deletion of a G base at position 2299 (g.2299delG) that results in a frameshift mutation. Expression analysis revealed high levels of IGF2BP2 mRNA in the liver, heart, and muscle tissues of female geese across embryonic (E25d), growing (A70d), and laying (L270d) stages, consistent with a potential role in muscle development (p < 0.05). Functionally, overexpression of IGF2BP2 in skeletal muscle satellite cells (SMSCs) was associated with significant changes in the expression of several genes linked to muscle development and signaling pathways, including upregulation of IGF1, EGFR, FGF19, BMP6, BMP2, ACVR1C and WNT5A and downregulation of MYBPC3, NODAL, HOXD13, TNXB, and ADD2 (Padj < 0.01). Furthermore, protein–protein interaction (PPI) network analysis of these genes suggests that IGF2BP2 may coordinate key genes, contributing to its potential role in skeletal muscle development in geese.

This experiment was designed to systematically evaluate the immunomodulatory effect of water extract of Artemisia annua L. (WEAA) on sheep, both in vivo and in vitro, and to determine the involvement of the TLR4/NF-κB signaling pathway in mediating these effects. In experiment 1, 32 female sheep (Dorper × Han, 3 months old, 24 ± 0.09 kg each) were designated to 4 groups, with each group receiving a basal diet supplemented with, respectively, 0 (control group), 500, 1000, and 1500 mg/kg WEAA. The serum, liver, and spleen immune indicators and related gene expressions were measured. In experiment 2, the peripheral blood lymphocytes (PBLs) were processed with WEAA (0, 25, 50, 100, 200, and 400 μg/mL), with six replicates assigned to each concentration group, then the cell viability, immune function, and related gene expressions were measured, and the optimal concentration of WEAA was determined. In experiment 3, the experimental groups consisted of PBLs subjected to treatments with or without PDTC (NF-κB inhibitor) and with or without WEAA, forming four distinct treatment groups (six replicates/group): PDTC(−)/WEAA(−) group, PDTC(−)/WEAA(+) group, PDTC(+)/WEAA(−) group and PDTC(+)/WEAA(+) group. The immune indexes and TLR4/NFκB pathway related indexes were determined. The results were as follows: WEAA dose-dependently enhanced the content of immunoglobulins (IgA, IgG, IgM) and cytokines (IL-1β, IL-2, IL-4) in the serum, liver, and spleen tissues, among which IgA, IgG, and IL-4 were the most significantly affected core indicators (p < 0.05). Meanwhile, WEAA dose-dependently upregulated the expression of TLR4/NF-κB pathway-related genes (TLR4, IKKβ, IκBα, NF-κBp65) and their downstream cytokine-related genes (IL-1β, IL-4) in liver and spleen tissues (p < 0.05). Of these genes, liver IL-4, IκBα, and spleen IL-4 were the most prominently regulated core genes (p < 0.05), The optimal supplementary dose of WEAA was determined to be 1000 mg/kg. In addition, adding 100 μg/mL WEAA to the culture medium of PBLs significantly enhanced immune function and cell viability. The underlying mechanism involved the TLR4/NF-κB pathway; that is to say, WEAA enhanced sheep’s immune indicators by upregulating TLR4/NF-κB pathway genes, thereby coordinately regulating humoral and innate immunity, thereby improving the immune indices of sheep. This study provided compelling experimental support for the prospective utilization of WEAA as a functional feed supplement in intensive meat-type sheep production systems.

Few studies have examined the effects of puerarin (PE) on ruminant parameters and methane production. Therefore, we determined the degradation of PE in the rumen and evaluated the effect of PE on in vitro fermentation, methanogenesis, and microbial community structure. A completely randomized design was used for the in vitro fermentation, and 4 gradient dosages of PE (0 mg/kg, 50 mg/kg, 100 mg/kg, and 150 mg/kg of DM) were applied in this trial. The in vitro fermentation was carried out in three runs at 6 h and 48 h, with four replicates per treatment per time point. Each run included 40 samples: eight treatments × four replicates and eight blank samples. Based on the fermentation results, both the PE treatments and the control group (CON) at 48 h were chosen for further analysis to explore the effects of PE on the bacterial community structure. Meanwhile, we determined the degradation rate and degradation products of PE in vitro ruminal fluid using high-performance liquid chromatography (HPLC). In this trial, PE may be isomerized into daidzin by rumen microorganisms; the in vitro degradation results of PE indicated that 70% of PE was degraded within 6 h, with the degradation rate reaching nearly 90% by 12 h. The concentrations of NH₃-N and microbial crude protein (MCP) significantly increased linearly with the PE doses at 6 h (p = 0.01). The concentrations of MCP (p = 0.02) and propionate (p = 0.04) demonstrated a linear increase with increasing PE doses at 48 h. In contrast to microbial protein (MCP) and propionate, the acetate-to-propionate ratio decreased linearly with increasing PE doses at 48 h (p = 0.05). Additionally, the addition of PE linearly decreased methane production at 48 h (p = 0.01). Meanwhile, the relative abundances of g_UBA1217 (p = 0.03), g_UBA2810 (p = 0.04), and g_Succiniclasticum (p = 0.03) were significantly lower compared with the CON group. The results showed that PE can be degraded by rumen microflora. Furthermore, it can improve rumen fermentation parameters, increase the amount of protein synthesized by rumen microorganisms, and reduce methane production and the acetate-to-propionate ratio. PE could potentially be an effective strategy for methane mitigation; however, further research is needed to assess its in vivo effects in dairy cows over a longer period.