Animal Science and Technology
Hoffman, Maria L
Animal and Veterinary Science
Fecal glucocorticoid metabolites; ELISA; rabbit
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The topic for this research project was based on the problem of the diminishing population of the wild New England Cottontail rabbits. Over the last 50 years, the population of these once very common rabbits has decreased to only inhabit New England and parts of New York (1). This is less than one-fifth of the geographic range they once inhabited (1). Roger Williams Park Zoo currently has a New England Cottontail breeding program that has experienced limited success, which may be caused by the domestic environment into which this wild species is being introduced. The central objective of this study was to determine the glucocorticoid metabolite concentrations in the fecal matter of established and newly obtained New England Cottontails using an enzyme linked immunosorbent assay. This would allow us to determine if the rabbits experience periods of increased stress during captivity. In order to determine the glucocorticoid metabolite concentrations, we started by using a protocol and antibodies purchased from the Smithsonian Institute (Front Royal, VA) which had been successfully used in species of pygmy rabbits. We also performed isolations with the fecal samples using ethanol, methanol, and water. Isolates were dried using fish tank aerators and fans to remove it. In optimizing this assay we determined that different buffer formulations were needed and amounts of reagents used in order to successfully coat the plate. Specifically, we determined that a protocol from Bethyl (Montgomery, Texas), along with new plates (Nunc-maxisorp plates; Thermofisher, Waltham, MA) coated much more efficiently. To date we have been able to successfully coat the plate and run hydrocortisone standards. However, we have found that the binding of the antibody appears to be non-specific. Therefore, we are going to attempt using a different block for the assay in an effort to determine if the issue is related to the blocking step or the primary antibody/secondary antibody concentrations. References: “New England Cottontail.” New England Cottontail | New England Cottontail Management, newenglandcottontail.org/.v