Methods:
Fifteen waterbodies in northern Michigan will be sampled. These sites include lakes, streams, and rivers in the surrounding area, including Douglas and Burt Lake. Twenty-five F. rusticus and 25 F. virilis will be collected from each site. Individuals will be captured using three minnow traps baited with sardines, and left for 24 hours. Traps will be placed at various habitat subtypes within the water body. All individuals captured will be transported back to the Experimental Stream Research Facility at the University of Michigan Biological Station (UMBS). Intraorbital length and propodus length (claw length) will be measured for each individual as well as sex and reproductive form. Crayfish will be housed by site in large tubs with flow through of untreated water from the East Branch of the Maple River, which runs adjacent to the facility. All sites and individuals will be physically and chemically isolated from each other. Individuals will be housed between June and August of 2024, in ambient temperatures and light and dark cycles for that time period. All individuals will be marked with white out to as a marker that will not impact behavior but allow for proper identification (Bergman & Moore, 2003, 2005) Crayfish will be fed natural detritus that flows in from the stream along with the water.
Environmental data collection will be conducted before traps are set at each site including, temperature, flow, turbidity, pH, and conductivity will be collected using a Hydrolab multimeter. Substrate composition will be determined by the dry mass percentage of substrate sizes. A kg of substrate will be removed from the site and allowed to dry for 24. Substrate will then separate by using multiple substrate sieve to get finally ratios of substrate types. In addition, macrophyte composition will be established using three 1-meter quadrats placed randomly around the collection sites. MacKay & Moore determined that the crayfish parasite Microphallus spp. could alter the behavior of infected hosts (MacKay & Moore, 2021). Due to this, post-experiment parasite analysis will be done by the dissection of the crayfish to remove the hepatopancreas for analysis. Boldness/shyness scale and aggression level assays will be run on all individuals. Crayfish will be run through two different behavioral assays, each day for three consecutive days. The order that crayfish will be run through the tests will be randomized.