Fatty acid composition plays a key role in regulating flavor and quality of milk. Therefore, in order to improve milk quality, it is particularly important to investigate regulatory mechanisms of milk fatty acid metabolism. Circular RNAs (circRNAs) regulate expression genes associated with several biological processes including fatty acid metabolism. In this study, high-throughput sequencing was used to detect differentially expressed genes in bovine mammary tissue at early lactation and peak lactation. Circ09863 profiles were influenced by the lactation stage. Functional studies in bovine mammary epithelial cells (BMECs) revealed that circ09863 promotes triglyceride (TAG) synthesis together with increased content of unsaturated fatty acids (C16:1 and C18:1). These results suggested that circ09863 is partly responsible for modulating fatty acid metabolism. Additionally, software prediction identified a miR-27a-3p binding site in the circ09863 sequence. Overexpression of miR-27a-3p in BMECs led to decreased TAG synthesis. However, overexpression of circ09863 (pcDNA-circ09863) in BMECs significantly reduced expression of miR-27a-3p and enhanced gene expression of fatty acid synthase (FASN), a target of miR-27a-3p. Overall, data suggest that circ09863 relieves the inhibitory effect of miR-27a-3p on FASN expression by binding miR-27a-3p and subsequently regulating TAG synthesis and fatty acid composition. Together, these mechanisms provide new research avenues and theoretical bases to improve milk quality.

The main purpose of this study was to explore the effect of tea tree oil (TTO) on lipopolysaccharide (LPS)-induced mastitis model using isolated bovine mammary epithelial cells (BMEC). This mastitis model was used to determine cellular responses to TTO and LPS on cellular cytotoxicity, mRNA abundance and cytokine production. High-throughput sequencing was used to select candidate genes, followed by functional evaluation of those genes. In the first experiment, LPS at a concentration of 200 µg/mL reduced cell proliferation, induced apoptosis and upregulated protein concentrations of tumor necrosis factor-α (TNF-α), interleukin 6 (IL-6), and signal transducer and activator of transcription 1 (STAT1). Addition of TTO led to reduced cellular apoptosis along with downregulated protein concentrations of nuclear factor kappa B, mitogen-activated protein kinase 4 (MAPK4) and caspase-3. In the second experiment, BMEC challenged with LPS had a total of 1,270 differentially expressed genes of which 787 were upregulated and 483 were downregulated. Differentially expressed genes included TNF-α, IL6, STAT1, and MAPK4. Overall, results showed that TTO (at least in vitro) has a protective effect against LPS-induced mastitis. Further in vivo research should be performed to determine strategies for using TTO for prevention and treatment of mastitis and improvement of milk quality.

We established a mastitis model using exogenous infection of the mammary gland of Chinese Holstein cows with Staphylococcus aureus and extracted total RNA from S. aureus-infected and healthy mammary quarters. Differential expression of genes due to mastitis was evaluated using Affymetrix technology and results revealed a total of 1230 differentially expressed mRNAs. A subset of affected genes was verified via Q-PCR and pathway analysis. In addition, Solexa high-throughput sequencing technology was used to analyze profiles of miRNA in infected and healthy quarters. These analyses revealed a total of 52 differentially expressed miRNAs. A subset of those results was verified via Q-PCR. Bioinformatics techniques were used to predict and analyze the correlations among differentially expressed miRNA and mRNA. Results revealed a total of 329 pairs of negatively associated miRNA/mRNA, with 31 upregulated pairs of mRNA and 298 downregulated pairs of mRNA. Differential expression of miR-15a and interleukin-1 receptor-associated kinase-like 2 (IRAK2), were evaluated by western blot and luciferase reporter assays. We conclude that miR-15a and miR-15a target genes (IRAK2) constitute potential miRNA–mRNA regulatory pairs for use as biomarkers to predict a mastitis response.

The composition of fatty acids plays a key role in regulating milk flavor and quality. Therefore, to improve the quality of milk, it is particularly important to study the regulatory...

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