Bile, a crucial fluid produced continuously by the liver, plays an essential role in digestion within the small intestine. Beyond its primary function in lipid digestion, bile also acts as a pathway for the elimination of various endogenous and exogenous substances. There have been limited studies focusing on interspecies differences. This study offers a comprehensive analysis of bile acid (BA) composition and its correlation with gene expression patterns across six different species, including mammals and poultry, through combining Liquid Chromatography-Mass Spectrometry (LC-MS) and transcriptome sequencing. The BA profiles revealed distinct metabolite clusters: D-glucuronic acid (GLCA) and glycochenodeoxycholic acid (GCDCA) were predominant in mammals, while taurolithocholic acid (TLCA) and T-alpha-MCA were prevalent in poultry, highlighting species-specific BA compositions. Differentially abundant metabolites, particularly GDCA, glycohyodeoxycholic acid (GHDCA) and taurodeoxycholic acid (TDCA) showed significant variations across species, with pigs showing the highest BA content. Transcriptome analysis of the liver and small intestine tissues of 56 cDNA libraries across the six species revealed distinct mRNA expression patterns. These patterns clustered samples into broad categories based on tissue type and phylogenetic relationships. Furthermore, the correlation between gene expression and BA content was examined, identifying the top 20 genes with significant associations. These genes potentially serve as biomarkers for BA regulation.

Background: During pregnancy, the immune status of maternal animals undergoes significant changes. Effective monitoring of immunological biomarkers can provide valuable references for enhancing gestational management in livestock. This study aims to screen and identify mRNA-based candidate immunological biomarkers during ovine pregnancy, thereby establishing a scientific foundation for novel monitoring technologies to assess immune status in gestating animals. Methods: High-throughput sequencing technology was used to perform sheep ovaries transcriptome sequencing in four stages: follicular phase, luteal phase (10 days after estrus), 30 days of pregnancy, and 45 days of pregnancy. Differential mRNAs were obtained by bioinformatics analysis. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses were performed on the differential mRNAs. Result: The experiment successfully constructed mRNA libraries for pregnant and non-pregnant sheep, with an average output of 11.50 G data per sample. The average alignment rate of each sample to the genome was 93.88%, and the average alignment rate to the gene set was 49.46%. A total of 23,219 genes were detected, of which 467 were differential genes. Data analysis, GO, and KEGG pathway studies showed that MX2, DDX58, IRF7, IFIT5, ISG15, RSAD2, and PLA2G1B were significantly upregulated in ovarian tissues during pregnancy, all of which are involved in processes such as bacterial and viral defense, immune regulation et al. QRT-PCR was used to detect the expression of seven different mRNAs in the blood of pregnant ewes, and the result confirmed that the expression was significantly elevated in the blood of pregnant ewes (p less than 0.01). The experiment indicates that the seven mRNAs are promising candidate biomarkers for immune monitoring, which is of great significance for the screening and early intervention of diseases during animal pregnancy.

Imidacloprid (IMI), a neonicotinoid insecticide, is widely recognized for its environmental persistence and toxicity to non-target aquatic organisms. Extended photoperiod exposure (EPE), an emerging anthropogenic stressor, further disrupts aquatic ecosystems by altering physiological and biological processes. However, their combined impacts on aquatic species remain insufficiently explored. This study evaluates the synergistic effects of IMI and EPE on Procambarus clarkii, an ecologically and economically significant crayfish species. Crayfish were exposed to 25 µg/L IMI under normal photoperiod (1000 lx, L:D = 12:12 h) and additional intensified and extended photoperiod (5000 lx, L:D = 18:6 h) treatments over one month. Key parameters, including survival rate, growth performance, oxidative stress markers, immune enzyme activities, neuroendocrine hormone levels, and gene expression, were assessed. The results indicate that EPE significantly amplifies the adverse effects of IMI. EPE reduced survival rates and growth performance, particularly in the 5000 lx group. IMI combined with EPE markedly elevated oxidative stress, as evidenced by increased malondialdehyde (MDA) levels and altered activities of superoxide dismutase (SOD) and catalase (CAT). Immune functions were impaired, with significant reductions in lysozyme (LZM) and acid phosphatase (ACP). Neuroendocrine disruption was observed through suppressed melatonin (MT) levels under EPE. Gene-expression analysis revealed upregulation of oxidative stress and apoptotic pathways (Cu/Zn-SOD, CAT and caspase-3) and downregulation of anti-apoptotic genes (bcl-2) and molt-inhibiting hormone (MIH). This study demonstrates that EPE exacerbates IMI-induced physiological and biochemical disruptions in P. clarkii. The findings highlight the pressing need for integrated management strategies addressing chemical and light pollution to protect aquatic ecosystems and sustain economically important species like crayfish.