EVALUATION OF CYTOCHROME P450-DERIVED LIPID METABOLITES AS THERAPEUTIC TARGETS DURING MASTITIS IN DAIRY CATTLE

<b>Project Methods</b><br> The project will be conducted using a combination of in-vitro experiments in the laboratory and an in-vivo model where animalswill experimentally challenged with Escherichia coli bacteria to reproduce the natural coliform mastitis condition. Theexperimental laboratory model will utilize cells derived from the vasculature of the bovine mammary gland to performexperiments in a controlled environment. Proven experimental methods in cell culture and state of the art analyses of samplesusing microscopy and liquid chromatography-tandem mass spectrometry, among other techniques, will be used. Data analyseswill be performed using a combination of parametric and non-parametric analyses and other suitable methods for researchtechniques utilized. The research methods will be valuable educational and practical experience training for graduate andundergraduate students working on the project.At the completion of the proposed studies, several milestones would have been achieved. First, the scientific underpinning ofhow the CYP450 pathway is altered during disease and how it correlates to disease severity disease will be demonstrated.Quantitative measures of gene expression and evidence of damage in in-vitro models will serve as measurable evidence of therelevance of the CYP450 pathway. Second, the manipulation of the CYP450 pathway will demonstrate quantifiable benefitsutilizing the in-vitro models. Based on the beneficial effects of this manipulation, demonstrable improvement in the clinicalseverity, disease progression, and clinical duration will be the confirmatory evidence that the CYP450 pathway is a viabletherapeutic intervention target. Ultimately, the specific manipulation will be tested to demonstrate the beneficial effects duringnaturally occurring coliform mastitis infection.