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Objective: It has been suggested that geographic variations in environmental toxins may impact sperm quality. Previously, we published findings that demonstrated a decline in sperm quality over an eleven year period across six regions of the United States (US).1 The current study investigates whether there was a decrease in sperm quality from a diverse set of US sperm donors across six regions in the US over a sixteen year period. Additionally, this study sought to evaluate changes between semen analysis (SA) parameters in an era of sperm collection during the COVID-19 pandemic. Materials and Methods: Semen analyses (SA) from sperm donors aged 19-38, with 2-5 days abstinence, from 9 different geographic regions from January 2005-April 2021 were examined. The sperm donors originated from one of the following regions: Palo Alto, Los Angeles, Westwood, International Nordic Cryo Bank Denmark, Indianapolis, Cambridge, New York, Houston, and Spokane, WA. Donation date, BMI, and geographic region were recorded. Data was analyzed as a whole as well as by individual region. Primary outcomes were ejaculate volume (mL), average concentration (M/mL), motility (%), and total motile count (M). Data was analyzed using a general estimate equation (GEE) model with an exchangeable working correlation structure to account for repeated measures. Results: A total of 176,706 SA specimens (from 3,532 unique donors) were analyzed. Controlling for BMI, there was a significant decline in average concentration (M/mL) (β=-1.89, p |