This study aimed to identify candidate genes regulating ovarian development following ovarian tissue transplantation through transcriptome sequencing (RNA-seq) and weighted gene co-expression network analysis (WGCNA). Ovarian tissues were collected from 10 thirty-day-old donor chickens, and each ovary was divided equally into three parts of similar size, and transplanted in situ into two-day-old recipient chickens. Samples were collected on days 0 (untransplanted), 6, and 12 after transplantation. RNA was extracted from ovarian samples for construction of a transcriptome library, and then sequenced on the IlluminaHiSeqTM2000 sequencing platform. The sequencing results were evaluated to determine differentially expressed genes (DEGs) through GO function and the KEGG pathway analyses, Gene Set Enrichment Analysis (GSEA), WGCNA, and PPI networks. Some candidate genes were further validated through real-time fluorescence quantitative PCR (RT-qPCR). RNA-seq analysis identified 2242 up-regulated and 3095 down-regulated DEGs at 6 days after ovarian transplantation, and 2181 up-regulated and 2129 down-regulated DEGs were identified at 12 days after transplantation. Enrichment analysis showed that the genes were enriched in multiple inflammatory pathways, pathways involved in signal transduction and cellular processes. Through the WGCNA, five modules were constructed, from which 4 and 5 candidate genes were mined at 6 and 12 days after ovarian transplantation, respectively. Transcription factor prediction showed that GTF3A was the most important transcription factor. The results of RT-qPCR verification confirmed that the expression profile of 8 candidate genes was consistent with that of the sequencing results. In summary, this study presents the candidate genes involved in ovarian vascular remodeling and ovarian proliferation after ovarian transplantation.

Gut microbiota plays an important role in animal health, production and diseases. Little is known about whether heat stress alters the composition of quail gut microbiota; therefore, we studied the effects of heat stress on growth performance, antioxidant functions, heat shock proteins and caecal microbiota. Two groups of 40 (20-day-old) quails were set up, including a control reared at 24 ± 2°C and a heat stress group subjected to heat stress at 36 ± 2°C for 4 h per day for 7 consecutive days. We found that heat stress significantly elevated the relative expression levels of HSP70 and HSP90 mRNA in the thymus, bursa and spleen by real-time fluorescence quantitative PCR assay, indicating successful establishment of the experimental model. Heat stress was found to have an effect on gut microbiota composition. At the genus level, Alistipes were significantly increased in the heat stress group. PICRUSt2 function prediction revealed that most of the KEGG pathways with high temperature-induced abundance differences are metabolism-related. These data indicated that heat stress reduced the production performance of quails by affecting antioxidant functions, as well as the composition and structure of the intestinal microbiota. The results of this study provide technical information for conducting research on heat stress prevention and control techniques in quails.

Beauvericin (BEA) is an emerging mycotoxin widely detected in feeds in recent years, posing a serious threat to the physiological health of dairy cows. Current studies on the biological toxicity of BEA mainly focus on the reproductive system, while research regarding mammary gland development in dairy cows remains scarce. Therefore, we used bovine mammary alveolar cells (MAC–T) as the research object to investigate the toxic effects of BEA and its underlying mechanisms. Our results demonstrated that BEA induced cell cycle arrest; with increasing doses, BEA altered cell morphology, stretched cell membranes toward both poles and triggered cell death. Further studies revealed that BEA exposure not only promoted intracellular reactive oxygen species (ROS) accumulation but also induced endoplasmic reticulum stress. BEA also suppressed the expression of genes associated with milk fat synthesis, including FASN, ACSS2, FABP3, SCD, DHCR7 and INSIG1. In addition, altered phosphorylation of JAK2/STAT3 and ERK1/2 was also observed following BEA exposure. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses showed that differentially expressed genes (DEGs) were significantly enriched in processes such as cell cycle, cytoskeleton organization and metabolism, which helped to characterize the toxic effects of BEA in bovine mammary epithelial cells.