The increasing dependence on amphibian captive breeding programs to support at-risk populations has led to growth in the application of assisted reproductive technologies (ART) to aid with the management of decreasing population density and genetic diversity. Already the integration of ART within these programs has led to monumental successes and shows great promise as a means to augment amphibian breeding efforts when natural breeding has low success. The focus of this project was to continue the optimization of ART to aid in making these technologies more widely applicable. Specific objectives include developing a framework that can be used to standardize follicular grading of oocytes through ultrasound assessments, demonstrating how advantageous routine sonography assessments are in providing insight into the female reproductive cycle in captive amphibians, and determining the differences in offspring growth and development between Ambystoma tigrinum larvae produced through in-vitro fertilization with either freshly collected or frozen-thawed sperm.