1. Presented by Rachel Barlow

2. According to the EAA Request for Proposals (RFP) for this study: Comal Springs riffle beetle (CSRB) distribution is currently unknown CSRB collection methods can be improved Per the RFP, this study is intended to: Establish a distribution of springs serving as CSRB habitat Test the success rate of at least three collection methods Collect CSRB for use in other 2014 Applied Research studies

4. Currently Understood CSRB Distribution within the Comal System Western shoreline Off northeastern tip of Spring Island Between western shoreline and northernmost peninsula of Pecan Island Spring runs 1-3

5. BIO-WEST 2002

7. Previous Collection Methods Fingers and/or soft forceps Dip nets Cotton cloth lures Cotton lures inside PVC tubing Drift net sampling Cotton cloth lure allowed to “culture” in situ for a period of four weeks

8. Study Objectives Test the success rate of three lure types Establish CSRB distribution within the Comal System

9. Methodology Preliminary trials with surrogate species Collect 250 CSRB (maximum) Store in aquaria Beetle Collection and Housing

10. Methodology (con’t) Four experimental chambers - Three test systems - Control Treatments (24 hour test cycle x 3) - Cotton cloth lure - Lighted cotton cloth lure - Hemp cloth lure 50 CSRB per chamber Expected results and analysis - % CSRB captured per test cycle - ANOVA (lure type) Laboratory Experimental Design

11. Methodology (con’t) Adaptive sampling - Only sample springs with CSRB habitat - CSRB present, move on Exclude springs with known CSRB Use most effective lure realized in laboratory study Survey (check traps) up to 3 times Survey no more than twice weekly Population Distribution Study

12. CSRB Habitat Criteria (based on physical parameters measured by Norris and Gibson 2013) Spring type: upwelling Temperature: 68-75°F Substrate consisting of organics, sand, course sand, very small gravel, small gravel, medium gravel, rubble, small cobble, and large cobble Flow sufficient to maintain clear substrate conditions

13. Expected Results Tested collection method Updated map of CSRB distribution Animals per trap across spring orifices Examination of other contributing parameters Number of surveys leading to presence/absence determination Nature of changing spring locations in relation to presence/absence studies