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Organisms in the wild have to allocate limited resources towards competing functions such as reproduction, growth, and self-maintenance. These competing investments create significant changes in physiological activity, and we still know little about the relationship between physiological activity and reproductive investment in natura. We investigated trade-offs between physiological activity and reproductive investment in the parthenogenetic Colorado checkered whiptail, Aspidoscelis neotesselata, across three different sites at the US Army Fort Carson Military Installation near Colorado Springs, CO, throughout the reproductive season in 2018 and 2019. We measured clutch size and reproductive activity and quantified plasma corticosterone (CORT), reactive oxygen metabolites (ROMs), and bacterial killing ability (BKA) to examine how energy-mobilizing hormones, oxidative stress, and immunity change in light of reproductive investment across different sub-populations. Circulating CORT increased with reproductive investment across all sub-populations, and increased clutch size led to a decrease in BKA in one sub-population, suggesting that habitat and nutritional availability may mediate this relationship. Oxidative stress, CORT, and innate immunity were not correlated with the exception of a trade-off between ROMs and BKA. This indicates individuals that have a better capacity to fight-off pathogens suffered increased reactive oxygen metabolites across all sub-populations, independently of habitat characteristics, which has important implications for A. neotesselata conservation. [ABSTRACT FROM AUTHOR]