Behaviors of Minnows (Cyprinidae) in Response to Alanine and Histidine in a Field Setting AquaVu underwater camera Plume to be measured Pipe to evenly release amino acid solution Katie Fessler, Department of Biology, York College Acknowledgments: I would like to thank Dr. Landis for allowing me to use all of his video equipment and devoting countless hours to helping me use the equipment out at the creek. I would like to thank Dr. Rehnberg for his guidance throughout this entire project. I would also like to thank Dr. Kleiner for disassembling the man-made dam in my test area, Dr. Thompson for the donation of his television, Christina Johnson and the remainder of the Biology Department faculty for all much needed help and encouragement! Introduction: Through gustation and olfaction, fish detect chemical cues in their environment that may cause behavior changes. Scientists test these abilities of fish in the laboratory by exposing them to amino acid solutions and identifying specific responses. The laboratory has controlled conditions at all times to ensure the behavior comes only from the administered solution. What happens in a natural stream? To answer this question, the minnows swimming in Tyler Run on the York College campus were presented with various concentrations of two amino acids: alanine and histidine. Past laboratory experiments have focused on catfish, trout, and zebrafish, whereas my research examined the reactions of minnows, including blacknose dace and creek chub. According to laboratory research, when trout detect alanine through smell, they are attracted to the source; when they detect histidine, they are repelled. Hypothesis Hypothesis: At high enough concentrations, minnows will show the same reactions as trout in a field setting. Alanine will cause an increase in the number of minnows within the field of view of the camera while histidine will cause an evacuation. Locate significant aggregation of minnows Place solution distribution pipe, Position camera, Administer red dye, Measure plume created Leave area undisturbed for 30 minutes and use creek water to make 10 -8, 10 -7, and M solutions of histidine or alanine 1. Videotape for 1 minute 2. Administer solution 3. Videotape for 3 minutes 4. Wait 15 minutes Order of solutions for Steps 1-4: a. 2 L creek water control b. 1 L M solution, 1 L creek water c. 1 L M solution, 1 L creek water d. 1 L M solution, 1 L creek water e. 2 L creek water control Repeat 12 times using alanine Repeat 8 times using histidine Analyze tapes and quantify behaviors Methods Methods : Test Subjects: Eastern Blacknose Dace (Rhinichthys atratulus) Creek Chub (Semolitus atromaculatus) Results: Conclusions: Behaviors Documented: 1. Number of minnows 2. Darting Considered an attracting odor if minnows dart toward solution Considered a repelling odor if minnows dart away from solution Alanine: According to paired t-tests, there was no difference between how the minnows reacted to the control in contrast to the alanine solution. Histidine According to paired t-tests, there was no difference between how the minnows reacted to the control in contrast to the histidine solution. Alanine vs. Histidine When comparing darting behavior and number of fish between alanine and histidine, there was no difference. Number of Minnows (%) Control vs M Concentration of Alanine Figure 1. Visually, it appears that the minnows were able to detect alanine, but a paired t-test calculated p=0.9399, meaning that there was no true increase in the number of minnows once the solution was administered. The number of minnows is in terms of percentage so it was possible to average all replicates regardless of the amount of fish tested each day. Number of Minnows (%) Control vs M Concentration of Histidine Figure 2. A paired t-test calculated p=0.0568, demonstrating that there was not quite a difference in the number of minnows during the M histidine test in contrast to the control test. The number of minnows is in terms of percentage so it was possible to average all replicates regardless of the amount of fish tested each day. Histidine vs. Alanine: Number of Minnows (%) at M Concentration Figure 3. A paired t-test calculated p=0.4322, demonstrating that there was no difference in the number of minnows during the M alanine test in contrast to the M histidine test. The number of minnows is in terms of percentage so it was possible to average all replicates regardless of the amount of fish tested each day. If the minnows could truly detect alanine or histidine, the many variables in a field setting were too overpowering to yield any significant statistical results.