Precision dietary interventions (e.g., altering proportions of dietary protein fractions) has significant implications for the efficiency of nutrient use in ruminants, as well as lowering their environmental footprint, specifically nitrogen (N) emissions. Soluble protein (SP) is defined as the protein fraction that is rapidly degraded in the rumen (e.g., non-protein N and true protein), and our previous study found that regulating SP levels could improve N efficiency in Hu sheep. Thus, the present study was conducted to explore in vitro how protein fractions with different SP levels modulate the rumen microbial community and its association with N metabolism. Four dietary treatments with different SP proportions and similar crude protein (CP) content (~14%) were formulated (% of CP): 20 (S20), 30 (S30), 40 (S40) and 50 (S50). Results showed that NH₃-N content increased with increasing SP levels at 4, 12 and 24 h; TVFA, acetate, propionate and valerate were higher in S30 and S40 (p < 0.05) and had quadratic effects (p < 0.05). Moreover, dry matter digestibility (DMD) and N digestibility (ND) were all decreased with S20 and S50 (p < 0.05). The S30 and S40 treatments increased the abundance of Bacteroidetes and Prevotella (Prevotella_ruminicola) but decreased the abundance of Firmicutes and Proteobacteria (p < 0.05). Bacterial pathways related to amino acid and fatty acid metabolism also were enriched with S30 and S40. The abundance of Entodinium was increased with S30 and S40 and had a positive correlation with Prevotella, and these two genera also played an important role in N metabolism and VFA synthesis of this study. In conclusion, bacterial and protozoal communities were altered by the level of SP (% of CP), with higher SP levels (~50% of CP) increasing the microbial diversity but being detrimental to rumen N metabolism.

Broccoli is a nutritious vegetable. It is high in protein, minerals, and vitamins. Also, it possesses antioxidant activities and is beneficial to the human body. Due to its active effect, broccoli is widely accepted by people in daily life. However, in terms of current utilization, only its florets are consumed as vegetables, while more than half of its stalks and leaves are not utilized. The stalks and leaves contain not only nutrients but also bioactive substances with physiologically regulating properties. Therefore research into the action and mechanism of its bioactive substances as well as its development and utilization technology will make contributions to the further promotion of its resource development and utilization. As a theoretical foundation for the resource utilization of broccoli stalks and leaves, this report will review the distribution and consumption of broccoli germplasm resources, the mechanism of action of bioactive substances, and innovative methods for their exploitation.

Background: RADIALIS (RAD), belongs to the MYB gene family and regulates a variety of functions including floral dorsoventral asymmetry in Antirrhinum majus and development of fruit proteins in Solanum lycopersicum. RAD genes contain an SNF2_N superfamily domain. Here, we comprehensively identified 68 RAD genes from six different species including Arabidopsis and five species of cotton. Results: Phylogenetic analysis classified RAD genes into five groups. Gene structure, protein motifs and conserved amino acid residues indicated that GhRAD genes were highly conserved during the evolutionary process. Chromosomal location information showed that GhRAD genes were distributed unevenly on different chromosomes. Collinearity and selection pressure analysis indicated RAD gene family expansion in G. hirsutum and G. barbadense with purifying selection pressure. Further, various growth and stress related promotor cis-acting elements were observed. Tissue specific expression level indicated that most GhRAD genes were highly expressed in roots and flowers (GhRAD2, GhRAD3, GhRAD4 and GhRAD11). Next, GhRAD genes were regulated by phytohormonal stresses (JA, BL and IAA). Moreover, Ghi-miRN1496, Ghi-miR1440, Ghi-miR2111b, Ghi-miR2950a, Ghi-miR390a, Ghi-miR390b and Ghi-miR7495 were the miRNAs targeting most of GhRAD genes. Conclusions: Our study revealed that RAD genes are evolutionary conserved and might be involved in different developmental processes and hormonal stress response. Data presented in our study could be used as the basis for future studies of RAD genes in cotton.

Background: This study aimed to compare three cotton-stalk processing strategies—grinding (FS), steam explosion (PH), and microbial fermentation (FJ)—and to clarify whether fermented cotton stalks preserve colonic epithelial integrity through a microbiota–metabolite–NF‑κB/MLCK axis in Hu sheep. Methods: Fifteen clinically healthy Hu sheep (26.7 ± 1.76 kg body weight; 115 ± 4 days of age) were used after a 14‑day adaptation period and randomly assigned to one of three diets ( n = 5 per treatment) containing 40% processed cotton stalks (FS, PH, or FJ) for 8 weeks. Results: PH and FJ increased final body weight compared with FS, and average daily gain increased progressively from FS to PH to FJ (206.07, 282.50, and 322.14 g/d, respectively; p < 0.05). Colonic fermentation profiles were markedly improved by FJ, evidenced by lower pH, ammonia nitrogen, free gossypol, and acetate ( p < 0.05), alongside higher total VFAs with elevated propionate and butyrate ( p < 0.05), whereas LPS was not different among treatments ( p > 0.05). Histology and scanning electron microscopy indicated that FJ maintained intact crypt architecture and epithelial surface continuity, while FS exhibited epithelial detachment and surface erosion. Metagenomic analysis revealed distinct community structures among groups, with FJ showing higher richness and enrichment of taxa associated with carbohydrate utilization and butyrate‑producing guilds (e.g., Lachnospiraceae ‑related genera such as Anaerostipes , Blautia , and Coprococcus ). Consistently, FJ suppressed colonic mucosal inflammation, as reflected by reduced IL‑1β, IL‑6, IL‑8, and TNF- α at both mRNA and protein levels ( p < 0.05). Mechanistically, FJ attenuated NF‑κB activation and downstream MLCK signaling, shown by decreased p‑p65/p65, p‑IκB/IκB, MLCK abundance, and p‑MLC/MLC ratio ( p < 0.05), while upregulating tight‑junction proteins (ZO‑1, occludin, claudin‑1, and claudin‑4; ( p < 0.05). Conclusion: Fermentation‑based processing of cotton stalks enhanced growth performance and promoted a favorable hindgut fermentation and microbial–metabolic milieu, thereby reinforcing colonic barrier integrity via inhibition of NF‑κB/MLCK‑associated inflammatory signaling, supporting fermented cotton stalk as a practical strategy to valorize cotton residues for ruminant feeding while mitigating gossypol‑related hindgut stress.

Aflatoxin B1 (AFB1) is a highly toxic mycotoxin which can be carried over into animal products and cause deterioration of livestock productivity when fed to livestock and wildlife. This review proposes a biomarker-guided framework for improving the early assessment of AFB1 exposure and toxicological responses in animal models. Oral exposure leads to the absorption of AFB1, which is bioactivated in the liver to the reactive AFB1-exo-8,9-epoxide that causes DNA and protein adduct formation, inflammation, mitochondrial apoptosis, and other effects. Cytochrome P450 activation and glutathione-dependent detoxification are in balance in determining susceptibility species, and this balance is different for poultry, pigs, ruminants, and rodents. In addition to traditional liver enzymes and histopathology, we highlight mechanistically informative biomarkers such as metabolites of aflatoxin, DNA and albumin adduct, lipid peroxidation products, antioxidant indices, cytokines, apoptotic markers, as well as signals involved in the Nrf2/NFκB pathway. AFB1 also damages the integrity of the intestinal barrier, the maintenance of the intestinal gut microbiota, reproductive function, growth performance, and development, thus creating a gut–liver-systemic toxic cascade. Finally, an assessment of stage-targeted interventions such as aluminosilicate binders, adsorbents derived from yeast, probiotics and nano-enabled interventions is conducted as viable tools for the reduction in exposure and injury. This review offers targeted mitigation strategies for early diagnosis of aflatoxicosis in animal production systems based on a biomarker approach.