Project description:Plant-derived secondary metabolites found in animal feed sources are beneficial for nutrition and health. Cowpea is a protein and phenolic-rich forage used as feed resource in animal system. The objective of this study was to understand the effect of cowpea secondary metabolites on gene expression in cows blood in vitro. Whole blood collected from Holstein Friesian cows (n=5) were treated with 10 ug/ml of cowpea leaf phenolic extract and untreated samples served as control. Total RNA was isolated and pooled together for microarray analysis. The Agilent one color bovine (v2) 4x44KÂ array was used and preliminary gene expression profiles generated using Cy3 labeled cDNA from CPE-stimulated and untreated samples. Gene expression analysis revealed a total of 3170 differentially expressed genes- 1716 up regulated and 1454 down regulated genes respectively. Pathway analysis identified CPE treatment association with innate immune response pathways including Toll-like receptor (TLR) signaling pathway, the Wnt signaling pathway, inflammation response pathway, and increased expression of the transcription factor NFKB1 were observed. Treatment with CPE decreased the expression of proinflammatory cytokines IL1A, IL1B and IL21. Quantitative real time PCR was performed to validate some gene members of the Toll-like receptor, inflammation response and Wnt signaling pathways. In vitro treatment with CPE impacted global gene expression profile in cow blood and results obtained in this study shows the potential immuno-modulatory properties of cowpea feed phenolic in cows. The global gene expression of the effect of cowpea phenolic extract (CPE) was measured in bovine peripheral blood.The experiment involved two groups; cowpea phenolic extract (CPE) treated samples vs untreated control group. Pooled RNA samples from each group was hybridized on Agilent one color bovine (v2) 4x44KÂ microarray slides. Two slides were prepared each with 4 array compartment.
Project description:Plant-derived secondary metabolites found in animal feed sources are beneficial for nutrition and health. Cowpea is a protein and phenolic-rich forage used as feed resource in animal system. The objective of this study was to understand the effect of cowpea secondary metabolites on gene expression in cows blood in vitro. Whole blood collected from Holstein Friesian cows were treated with 10 ug/ml of cowpea leaf phenolic extract and untreated samples served as control. Total RNA was isolated and pooled together for microarray analysis. The Agilent one color bovine (v2) 4x44K array was used and preliminary gene expression profiles generated using Cy3 labeled cDNA from CPE-stimulated and untreated samples. Gene expression analysis revealed a total of 3170 differentially expressed genes- 1716 up regulated and 1454 down regulated genes respectively. Pathway analysis identified CPE treatment association with innate immune response pathways including Toll-like receptor (TLR) signaling pathway, the Wnt signaling pathway, inflammation response pathway, and increased expression of the transcription factor NFKB1 were observed. Treatment with CPE decreased the expression of proinflammatory cytokines IL1A, IL1B and IL21. Quantitative real time PCR was performed to validate some gene members of the Toll-like receptor, inflammation response and Wnt signaling pathways. In vitro treatment with CPE impacted global gene expression profile in cow blood and results obtained in this study shows the potential immuno-modulatory properties of cowpea feed phenolic in cows.
Project description:Yamoa⢠is marketed and sold as a dietary supplement with anecdotal positive effects in asthma and hay fever. We determined that Yamoa⢠(ground bark of Funtumia elastica tree) stimulated innate immunity in part by affecting gamma delta T cells. Yamoa⢠had distinct priming effects, very similar to, but more robust than, that of lipopolysaccharide (LPS), on bovine, mouse and human gamma delta T cells. However, the optimal effect was dependent on the presence of accessory cells. Gene expression patterns in bovine gamma delta T cells and monocytes induced by Yamoa⢠were very similar to those induced by ultrapure LPS, but the agonists in Yamoa⢠did not signal entirely through TLR4. Yamoa⢠stimulated human cells to produce cytokines involved innate protection. The bioactive component of Yamoa⢠was delineated to a complex polysaccharide fraction (Yam-I). Intraperitoneal injection of Yamoa⢠and very low doses of Yam-I in mice induced rapid increases peritoneal neutrophils directed by changes chemokine expression. Yamoa⢠and Yam-I were effective as therapeutic treatments in mice with Salmonella enterica serotype Typhimurium (ST) induced enterocolitis that resulted in decreased bacterial counts in feces. This initial characterization of the immune stimulatory properties of polysaccharides derived from Yamoa⢠suggests potential mechanisms for positive effects in asthma and that they have potential for application in infectious disease settings. . Experiment Overall Design: To begin to understand the effects of Yamoa in innate immunity, we investigated the global gene expression profiles of stimulated bovine gamma delta T cells. Peripheral blood from 3 neonatal bovine calves was collected. gamma delta T cells were sorted to >97% purity using a FACS Vantage. Cells were placed in culture and stimulated with either an aqueous extract of Yamoa (32.6ug/ml), ultrapure LPS [uLPS (10ug/ml)] or PBS for 4 hours after which RNA was extracted and processed for microarray analysis.
Project description:Microarray-based gene expression profiling of peripheral blood mononuclear cells in dairy cows with experimental hypocalcaemia and milk fever
Project description:In vitro maturation (IVM) of the oocytes is a routine method in bovine embryo production. The competence of bovine oocytes to develop into embryo after IVM and in vitro fertilization (IVF) is lower as compared to in vivo preovulatory oocytes. Cumulus cells (CC) that enclose an oocyte are involved in the acquisition of oocyte quality during maturation. Using transcriptomic approach we compared cumulus cells gene expression during IVM with that in vivo preovulatory period.
Project description:Affymetrix Human GeneChips are used to profile gene expression of bovine tissues and embryos to identify uniquely expressed genes in bovine in-vitro fertilized embryos by comparing with seven bovine adult tissues through gene clustering