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Bio 213 Name:_______________________ Lab 9: Leaf litter Invertebrate Biodiversity INTRODUCTION In this field study we will quantify the diversity of the invertebrate communities from leaf litter samples. Diversity is a measure of the relative representation of species in a community. It is comprised of two components, species richness (the number of species) and species evenness (the relative number of individuals of each species). Thus, two communities may have the same number of species (richness) but one community may have one species that is more numerous than the others (evenness) (Table 1). The second community would be less diverse. This makes intuitive sense, since the second community would look to us as though it were composed of almost totally one species and would not look diverse. diversity. Table 1. A comparison of two communities and their apparent Number of Individuals Community 1 Community 2 20 92 20 2 20 2 20 2 20 2 100 100 Species A B C D E TOTAL There are several ways to quantify diversity. One is simply to calculate species richness. However, as illustrated in Table 1, this can give a false impression of the relative diversity of two different areas. Therefore, other measures have been devised by community ecologists. The one we will use is called Simpson’s index. Simpson’s diversity index = D= N (N − 1) ∑ ni (ni − 1) where N = total number of individuals of all species in a community and ni = number of individuals of the ith species or taxon. The summation symbol (∑) means to do the calculation following the ∑ for each of the species or taxa and then add up the results of all the calculations. D ranges from 1, for a community made up of one species (or other taxon), to infinity, for a community made up of one individual of many species (or other taxon). Contrary to its outward appearance to the casual observer, leaf litter is a dynamically active living community. The organisms are busy using leaf litter as a place to feed, reproduce, compete…live! In the process, they work the leaf litter into the soil, making it more fertile, improving its water-holding capability, increasing the ability of oxygen to enter the soil, and decreasing the soil's susceptibility to erosion. They therefore play an important role in the formation of soil. The organisms in leaf litter interact with one another and with the environment, and thus make up an ecological community. Energy is transferred through the community, and nutrients are recycled. There are predators, prey, decomposers, competitors, symbiotic partners. Plant material that drops to the soil surface, called plant or leaf litter, as well as the bodies of dead organisms, is collectively called detritus, and becomes food for various organisms called detritivores. They chew it up into smaller pieces, and leave fecal wastes that then become food for other organisms. Some of these organisms are eaten by predators. The detritus gets worked to smaller and smaller particles, and bacteria and fungi, the decomposers, and chemical action reduce the organic humus to the minerals that are nutrients used by plants. And the cycle continues. In this laboratory exercise, we will collect leaf litter animals and study this complex and highly intriguing community. This exercise will take several days to complete. OBJECTIVES In this study, you will a) identify various groups of animals that live in leaf litter. b) explore concepts of ecosystem ecology. DAY ONE: SAMPLE COLLECTION MATERIALS Burlese funnel system specimen vials, with alcohol bucket for leaf litter trowel PREPARATIONS 1. In class, we will determine where we want to sample for leaf litter invertebrates. We will choose several sites for analysis, and several groups will collect samples from the same locations. FIELD PROCEDURE 1. Work in groups of 3-4. 2. Get a trowel and bucket and go to collection site. 3. Write a description of the site on Tables 1 & 2. Be sure to include what kind of plants the leaf litter is coming from, light conditions and moisture conditions. 4. Using the trowel, dig into the leaf litter. You want to take samples from the bottom-most layer where the most decomposition is occurring, without getting soil. 5. Return to the lab and label your Burlese funnel with your site numbers and group name. 6. Place your Site 1 sample in the Burlese funnel apparatus marked Site 1. Repeat for Site 2. 7. Adjust the lamp so it is about 3 inches above your funnels and turn on the light. (Depending on the set up, this may be already done.) 8. Label a vial “Site 1” and another “Site 2”, fill about 3/4 with alcohol, and put each under the appropriate funnel. 9. Let the apparatus sit for several days. DAY TWO: DATA COLLECTION MATERIALS dissecting microscope squirt bottle of alcohol specimen vials, with alcohol Guide to common leaf litter and soil animals dissecting probes/needles top or bottom of petri dish Each group will be responsible for analyzing the contents of the two sites they collected. Note that some of these samples will be easier to analyze than others. If you are done early, help your classmates out! We cannot finish this exercise until we have the data from every group. 1. Turn off the light of the Burlese apparatus. 2. Obtain your sample vials (one from each site). 3. Empty the funnels in the trash, but be sure to save the pieces of wire mesh!! 4. Obtain a dissecting microscope, the top or bottom of a petri plate for each member of your group, and one or two dissecting needles/probes for each member. 5. Work with one vial at a time. Swirl the vial, then pour the contents into enough petri plates so that each member of your group can be working to identify and count the organisms. If you need to, use the alcohol squirt bottles to wash out any organisms that remain in the sample vial. 6. Place the petri plate under the microscope and sort the organisms by moving similar looking organisms together in groups with the dissecting needle/probe. Start at low power and search for organisms. Finish at high power, to be sure you have not missed anything. 7. Use the guide to identify the groups. 8. Count the number of individuals in each group, using "tick marks" in Tables 1 & 2 to keep track. a. A helpful way to do this is have one group member scan the petri dish and call out the identity of organisms they see (e.g., “pseudoscorpion, mite, mite, springtail, . . .”), while another group member keeps track of them by using tick marks. Count up the tick marks when finished. 9. After you have finished, add up the total number of individuals for each type of organism you found. Note that if your sample was divided into multiple funnels, you will need to pool your data with another group before determining the Simpson’s Index. 10. Repeat for Site 2. DATA & ANALYSIS Diversity: You and your lab mates will select two of the sampled communities to compare diversity. 1. Using the information above and your class data, calculate Simpson’s diversity index for each of the two communities you have chosen to compare. Use 2 decimal places. 2. D= Community 1: D= Community 2: 3. Which community is more diverse? How would you explain your results: Why might the one community be more diverse than the other? 4. Compare your results to your classmates. Of the various factors we tested, which was the best indicator of biodiversity in leaf litter communities? Why? ---------------------------------------------Sample calculation for Simpson’s Diversity Index Species Number (n) n(n-1) Woodrush 2 2 Holly (seedlings) 8 56 Bramble 1 0 Yorkshire Fog Sedge 1 3 0 6 Total (N) 15 64 Putting the figures into the formula for Simpson's Index Table 1: Site 1 data Description of Site 1: Type of Animal Thysanura (bristletails) Collembola (springtails) Thysanoptera (thrips) Protura Pseudoscorpions Ticks Mites Centipedes Millipedes Spiders Insect larvae Beetles Nematode worms Others Tick Marks (keep track of numbers here) Total Number Table 2: Site 2 data Description of Site 2: Type of Animal Tick Marks (keep track of numbers here) Total Number Thysanura (bristletails) Collembola (springtails) Thysanoptera (thrips) Protura Pseudoscorpions Ticks Mites Centipedes Millipedes Spiders Insect larvae Beetles Nematode worms Others REFERENCES (FOR FURTHER INFORMATION) Crossley, D. A. Jr. 1977. Pp. 49-56. In: W.J. Mattson (ed.) The role of arthropods in forest ecosystems. Springer-Verlag, New York. Edwards, C.A., D.E. Reichle, and D.A. Crossley, Jr. 1970. Pp. 147-172. In: D.E. Reichle (ed.) Analysis of temperate forest ecosystems. Springer-Verlag, New York. Petersen, H. and M. Luxton. 1982. A comparative analysis of soil fauna populations and their role in decomposition processes. Oikos 39: 287-388. Luff, M.L. 1975. Some features influencing the efficiency of pitfall traps. Oecologia 19:345-357. Smith, R.E. [ecology text]