How does heavy metal contamination affect reproductive parameters in Florida lizards?

Methods

Summary

Field Work

Field work will begin in Summer, 2021. 60 lizards of each species will be tested from each
sampling site (30 female and 30 male). Blood and tissue samples will be taken in the field. All
proposed sampling methods and handling/housing procedures have been approved by IACUC. Each lizard will be weighed with Pesola scale (g), sexed, and measured for total length and snout-to-vent length with a digital caliper (mm). Soil from each location will also be collected. Green anoles will be released immediately after data collection at the site of capture. Brown anoles will be taken back to University of South Florida St. Petersburg (USFSP) for further laboratory experiments. Lizards used in laboratory experiments will be kept in terrariums containing soil from their wild locations. 

Laboratory Experiments

Eight brown anoles (Four adult males and four adult females) from each site will be taken to the University of South Florida St. Petersburg for laboratory experiments. Captured lizards will be weighed, measured, and sexed. Body condition will be based off a body mass/snout-vent length ratio. Individuals will be tested for heavy metals within their blood and tissue samples, and males will have their testosterone levels tested. The metals tested for will be As, Cd, Pb, and Hg. Blood and tissue samples will be sent to Michigan State University Diagnostic Veterinary Laboratory for analysis. Lizards will be separated by level of urbanization. Each terrarium will house one male and one female lizard. The lizards will be bred, and their health and body conditions monitored. The total number of eggs from each brood will be counted, and hatchling survival rate will be recorded. The sex of the juveniles will also be determined, to monitor for skewed sex ratios across the various study sites. Growth rates of the hatchlings (length, weight) will be monitored, to test if/how differing levels of heavy metal contamination affects juvenile development. Once the juveniles reach maturity, they will be tested for heavy metal contamination using the same process of extracting blood and tissue samples, to test for inherited contamination.