Tardigrades of Harvard Forest: A Preliminary Report Mehlan R. New Tardigrada from Texas. American Midland Naturalist. 1969 April 81(2):395 Meyer HA. Terrestrial and freshwater Tardigrades of the Americas. Zootaxa. 2013 Dec 3747(1): 1-71 McInnes S.J. Zoogeographic distribution of Terrestrial/freshwater tardigrades from current literature. Journal of Natural History. 1994 April 28(2):257-352 Haefke B.J., Spiers A.I., Miller W.R.. & Lowman M.D. Tardigrades in the Canopy: Using Double Rope Techniques to Conduct Sampling Along Vertical Transects. Transactions of the Kansas Academy of Science. 2013 116(3-4): 119-124. Miller W.R. Bears of the Moss. The Kansas School Naturalist. 1997 May 43(3):16. Poster by Matthew Sears, Andrew Emanuels, and Irvin Chapa – Principle Investigators: William R. Miller, Ph.D. and Meg D. Lowman, Ph.D. Introduction Results Tardigrades are microscopic limno-terrestrial invertebrates that are found on all seven continents in a variety of substrates, like moss and lichen. Wide-scale distribution of tardigrades is an understudied subject, particularly in the eastern deciduous forests of Massachusetts. Previously, the state only had records of two species (Mehlan 1969; McInnes 1994; Meyer 2013). Tardigrades were collected from the canopies of a wide array of trees through Baker University’s Research Experience for Undergraduate (REU): “Tardigrades in the Canopy” funded by the National Science Foundation. The composition of the species of those trees were used to represent North America’s eastern deciduous forest biome. Over 5000 tardigrades were extracted from the canopy samples. Five types of trees were sampled: Red Oak, Beech, Birch, Sugar Maple, and White Ash. Sample height ranged from 0 to 16.5 meters. Sample pH values ranged from 2.1 to 7.14. Fifteen species of tardigrades were found in the canopies, representing 11 genera. Climbing picture Figure 1: Climbing Technique Double Rope Technique in the canopy, as seen from the ground. Pictured: Daniel Rivera Table 1 (left): Genera Found in Harvard Forest * = a new record for Massachusetts. ᶧ = contains a novelty species Table 2 (below): Density (tardigrades per sample) and pH Averages Methods Figure 2: SEM Picture of Minibiotus Taken by a Scanning Electron Microscope at Kansas University by the REU team Climbers worked in pairs and utilized the Double Rope Technique (Haefke et al., 2013). One hundred and three trees were climbed and collected from within six different sites in Harvard Forest (Figure 3). Samples were soaked in spring water for 24 hours (Miller 1997). Tardigrades found in each sample under a dissecting scope were mounted on slides in polyvinyl alcohol. pH was also taken of the samples. The animals were then identified to species level under a differential interference microscope (400x magnification). Cover slips were sealed on slides with fingernail polish 3-5 days later. Discussion This study expands the known Massachusetts tardigrade species diversity by 650% from 2 to 15. There was no significant difference in densities between levels. Numerous eggs and individuals in simplex and undergoing ecdysis were found, indicating a thriving and growing population. These findings broaden the scientific field’s knowledge of tardigrade distribution not only up and along the trunk of a tree, but also on a national scale. A B 100 µm 1 2 C 10 µm Figure 3: Harvard Forest Site Locations Circle in red are sites that were sampled from within Harvard Forest, accompanied with their respective GPS coordinates. Map provided by Harvard Forest Field Station’s website. 100 µm 3 4 Miller W.R. Bears of the Moss. The Kansas School Naturalist. 1997 May 43(3):16. Mehlan R. New Tardigrada from Texas. American Midland Naturalist. 1969 April 81(2):395 Meyer HA. Terrestrial and freshwater Tardigrades of the Americas. Zootaxa. 2013 Dec 3747(1): 1-71 McInnes S.J. Zoogeographic distribution of Terrestrial/freshwater tardigrades from current literature. Journal of Natural History. 1994 April 28(2):257-352 Haefke B.J., Spiers A.I., Miller W.R.. & Lowman M.D. Tardigrades in the Canopy: Using Double Rope Techniques to Conduct Sampling Along Vertical Transects. Transactions of the Kansas Academy of Science. 2013 116(3-4): 119-124. Funded by NSF:REU #1461005