1. Tardigrade Identification and Distribution at York College using Molecular Techniques Jason Lamontagne, York College of PA, Dept. of Biological Sciences Introduction Tardigrades, or water bears as they are known, are very unique animals. They are microscopic animals with digestive, excretory, nervous, and muscle systems, as well as four sets of legs, each ending in a set of finger-like claws (Miller 1997, Romano 2003). What makes them so unique, however, is their ability to survive extreme environments. They go through a process called cryptobiosis, during which a tardigrade will enter a reduced metabolic state in response to an environmental change, and they will stay in this state until environmental conditions become favorable again (Jönsson and Bertolani 2001). Research has shown that a tardigrade can go through anhydrobiosis (in response to lack of water) and be revived successfully after a decade (Guidetti and Jönsson 2002). Despite all of their interesting characteristics very little is known about the tardigrades and the processes they go through to enter these states. The biggest challenge, by far, in doing a tardigrade study is the lack of information on the subject. Without the help of an expert and very high magnification identification is nearly impossible. Most of the research published on tardigrades has been distribution studies (e.g. Jönsson 2003, Uhĩa and Briones 2002), which unfortunately does not help in the actual identification of the species in our area. To date, no known distribution studies have been done in the York area, despite proximity to such potentially environmentally rich areas as the Susquehanna River and Lake Redman. Because of these gaps in knowledge, this research proposes the development of a molecular identification technique using the 18S rDNA, and the application of this technique in a distribution study of tardigrades found on the York College campus. Hypotheses Part 1: The differences in the 18S rDNA will give an effective method for identification of tardigrade species Part 2: Tardigrades will not demonstrate a preference for certain moss species Figure 1. The various responses to environmental extremes that make tardigrades a very unique animal. Results Proposed Overall Conclusions The molecular identification technique used in this research showed the potential to be a viable alternative for identifying tardigrades. We feel that by optimizing the technique we would be able to differentiate among species. Similar techniques are being applied to other animals, which supports the hypothesis that this technique is a possible identification procedure. Once the identification technique is optimized, its application to a distribution study on the York College campus will prove its value in the field. Without having to rely on the help of an expert, studies will be completed faster and more easily. Methods Acknowledgments I would like to thank my research mentor Dr. Nolan for all of her help and patience. Also, thanks to many of the other members of the Biology faculty for helping me along the way with various parts of this research. Results Achieved Fig. 2Fig. 3 Fig. 2- The gel shows the final PCR products which were excised and sent out for sequencing. Fig. 3- The tree shows the species relations based on their 18S gene sequences. Fig. 5 Fig. 4- The graph shows a theoretical relationship between tardigrades and five moss species, showing preference for two specific species. Fig. 5- A tree showing the desired species distribution after optimization of the identification technique.