The two objectives for this thesis were to 1) validate a bovine luteinizing hormone (LH) ELISA for use in the laboratory and 2) validate interstitial glucose sensors designed for humans for use in lactating dairy cattle. The first experiment required validation of a bovine LH ELISA in order to measure the circulating concentration of LH in the blood of cows in altered thermal and/or metabolic states. Assays from two separate companies were tested. Half of one plate was run at a time resulting in a total of 6 separate analyses (3 plates total). Despite early, promising results, neither LH ELISA could be successfully validated. For many analyses, the standard curves did not even meet the minimal criteria to allow calculation of a formula for determining the concentrations of the unknowns. In analyses where the standard curves were acceptable, the coefficients of variation (CV%) for the unknowns were unacceptable. The second experiment attempted to validate the use of commercially available human interstitial glucose sensors (FreeStyle Libre and Dexcom G6) in lactating dairy cows. Blood glucose concentrations correlated well with sensors secured behind the cow's ear for both FreeStyle Libre (r=0.82) and Dexcom (r=0.88). Unfortunately, however, the absolute relative error's highest value was 47% for the FreeStyle Libre ear. In summary, neither the LH ELISAs nor the interstitial glucose sensors could be validated. / Master of Science / Two separate studies were performed with the purpose of one, measuring a hormone significant to reproduction, Luteinizing Hormone (LH), and two, measuring glucose in interstitial fluid in lactating cows. The first experiment required the use of a commercially available ELISA test to measure LH concentrations. While using radioimmunoassay (RIA) is the preferred method for measuring LH in dairy cattle, an ELISA is an alternative method that is generally more accessible. There was also little published research available illustrating the accuracy of the LH ELISAs. Therefore, we attempted to validate the assays for bovine samples. Despite several attempts, the LH ELISA was unsuccessfully validated. The second study attempted to validate commercially available continuous interstitial glucose sensors designed for humans for use with lactating Holstein cows. Validation of interstitial sensors for use in dairy cattle would decrease the amount of stress and handling when measuring circulating glucose for research, clinical diagnostics and application in industry. While early data suggested that sensors behind a cow's ear may work, further analysis illustrated the sensors were not accurate enough when used on cows. In conclusion, the lab was unsuccessful in proving that continuous interstitial glucose monitors and LH ELISAs could be utilized in the lab.
| Identifer | oai:union.ndltd.org:VTETD/oai:vtechworks.lib.vt.edu:10919/104678 |
| Date | 18 August 2021 |
| Creators | Harrod, Mary Kathryn |
| Contributors | Animal and Poultry Sciences, Rhoads, Michelle, Ealy, Alan Dale, Mercadante, Vitor R. G. |
| Publisher | Virginia Tech |
| Source Sets | Virginia Tech Theses and Dissertation |
| Detected Language | English |
| Type | Thesis |
| Format | ETD, application/pdf |
| Rights | In Copyright, http://rightsstatements.org/vocab/InC/1.0/ |
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