1. Comparative Analysis of Eisenia Movement In Increasing Compost Concentrations Matthew M. Steffenson Masters Graduate Student Tennessee Technological University Department of Biology Cookeville, TN 38505

2. Executive Summary Objectives and Goals To identify methods of testing Eisenia mortality in different compost concentrations. To identify methods of testing Eisenia mortality in different compost concentrations. Determine Eisenia behavioral choices of habitat based upon physiological tolerances. Determine Eisenia behavioral choices of habitat based upon physiological tolerances.

3. Executive Summary Hypothesis and Expected Results Alternative: Earthworms will choose lower concentrations of compost to minimize interaction with high levels of acidic nutrients/compounds. Additionally, earthworms with a higher body-mass index will be found in higher concentrations of compost. Alternative: Earthworms will choose lower concentrations of compost to minimize interaction with high levels of acidic nutrients/compounds. Additionally, earthworms with a higher body-mass index will be found in higher concentrations of compost. Null: The earthworms will be found in equal numbers in all compost concentrations, and body-mass index will not differ significantly in any concentration. Null: The earthworms will be found in equal numbers in all compost concentrations, and body-mass index will not differ significantly in any concentration.

4. Executive Summary Procedural Outline Earthworms will be place in a compost trough with an increasing gradient of concentrations. Earthworms will be place in a compost trough with an increasing gradient of concentrations. After a predetermined period of time, worms will be captured and length, mass and location will be recorded. After a predetermined period of time, worms will be captured and length, mass and location will be recorded.

5. Introduction Presence or absence of earthworms within a population can be an indicator of the health of the soil (Deibert 1997). Presence or absence of earthworms within a population can be an indicator of the health of the soil (Deibert 1997). In bioremediated fields, earthworm lethality assays have been found to be the most sensitive methods to assess soil quality (Dorn and Salanitro 2000). In bioremediated fields, earthworm lethality assays have been found to be the most sensitive methods to assess soil quality (Dorn and Salanitro 2000). Some vineyards add copper fungicides to their soil. Found that this has a negative impact on growth and survival of earthworms. In turn, may lead to decreased soil quality (Eijsackers et al. 2005). Some vineyards add copper fungicides to their soil. Found that this has a negative impact on growth and survival of earthworms. In turn, may lead to decreased soil quality (Eijsackers et al. 2005). Additionally, earthworm presence has been found to have a positive linear relationship with soil minerals and microbial biomass as well as crop yield (Eriksen-Hamel and Whalen 2006). Additionally, earthworm presence has been found to have a positive linear relationship with soil minerals and microbial biomass as well as crop yield (Eriksen-Hamel and Whalen 2006).

6. Introduction Statement of Problem: (1) Do earthworms choose habitats that have better soil quality and; (2) are earthworms with a larger body-mass index capable of surviving in higher concentrations of compost? Statement of Problem: (1) Do earthworms choose habitats that have better soil quality and; (2) are earthworms with a larger body-mass index capable of surviving in higher concentrations of compost? Hypothesize that earthworms with a larger body- mass index will be found in higher concentrated areas and that mortality will be low due to behavioral choices. Hypothesize that earthworms with a larger body- mass index will be found in higher concentrated areas and that mortality will be low due to behavioral choices. Null hypothesis states that an equal number of earthworms will be found in each concentration of compost, and that length/mass will be randomly distributed. Null hypothesis states that an equal number of earthworms will be found in each concentration of compost, and that length/mass will be randomly distributed.

7. Methods Experimental chamber will be made out of plywood. Experimental chamber will be made out of plywood. Dimensions (.20m deep Eriksen-Hamel and Whalen 2007):Dimensions (.20m deep Eriksen-Hamel and Whalen 2007): Testing chamber will be lined with plastic and then have holes drilled in for aeration. Testing chamber will be lined with plastic and then have holes drilled in for aeration. Chamber will then be split lengthwise to allow for 2 repetitions, A and B. Chamber will then be split lengthwise to allow for 2 repetitions, A and B. Chamber will be separated into 4 sections (Control, 25%, 50%, 75%) using plastic mesh. Chamber will be separated into 4 sections (Control, 25%, 50%, 75%) using plastic mesh. Compost and control soil will be obtained at the Tech Waters Organic Farm. Compost and control soil will be obtained at the Tech Waters Organic Farm. Compost will be mixed with a base soil type (control) at the aforementioned percentages by volume.Compost will be mixed with a base soil type (control) at the aforementioned percentages by volume. Earthworms will be placed in control soil and allowed to acclimate for 1-3 days. Earthworms will be placed in control soil and allowed to acclimate for 1-3 days. Temperature will be held constant at 20 degrees Celsius (Nahmani et al. 2007; Gibbs et al. 1996), and moisture will be held at approximately 20% (Eijsackers et al. 2005). Temperature will be held constant at 20 degrees Celsius (Nahmani et al. 2007; Gibbs et al. 1996), and moisture will be held at approximately 20% (Eijsackers et al. 2005). All earthworms (120) will be placed into the control at the beginning of the experiment. All earthworms (120) will be placed into the control at the beginning of the experiment. Earthworms used will be Eisenia hortensis. Earthworms used will be Eisenia hortensis. After 14 days the earthworms will be collected (Dorn and Salanitro 2000; Gibbs et al. 1996). After 14 days the earthworms will be collected (Dorn and Salanitro 2000; Gibbs et al. 1996). Location in chamber, length, and mortality will be recorded. Location in chamber, length, and mortality will be recorded. Test will be conducted at the Hyder-Burks greenhouse. Test will be conducted at the Hyder-Burks greenhouse. 0.5m 1.0m Control25%50%75%

8. Methods Data Analysis Location Data: Location Data: Concentration gradients will be compared using a Fisher’s exact t-test.Concentration gradients will be compared using a Fisher’s exact t-test. Mortality/Length Data: Mortality/Length Data: Mortality data will be compared using a Chi- squared.Mortality data will be compared using a Chi- squared. Concentration gradients will be compared within troughs using one-way ANOVA’s with a Tukey’s post-hoc (Eijsackers et al. 2005).Concentration gradients will be compared within troughs using one-way ANOVA’s with a Tukey’s post-hoc (Eijsackers et al. 2005). All data will be analyzed using Statistical Package for the Social Sciences (SPSS) software. All data will be analyzed using Statistical Package for the Social Sciences (SPSS) software.

9. Expected Results and Benefits Predict that larger numbers of earthworms will be found in lower concentrations of chicken compost. Predict that larger numbers of earthworms will be found in lower concentrations of chicken compost. Additionally, mortality be minimal (i.e. habitat choice). Additionally, mortality be minimal (i.e. habitat choice). Also predict that earthworms found in higher concentrations of compost will have a higher body-mass index (i.e. physiological tolerances). Also predict that earthworms found in higher concentrations of compost will have a higher body-mass index (i.e. physiological tolerances). Can use results to identify tolerances of earthworms. Can use results to identify tolerances of earthworms. Results will act as a marker to quantify compost concentrations at which plants will grow best by reason of presence of both nutrients from fertilizer and tillage/aeration due to earthworms. Results will act as a marker to quantify compost concentrations at which plants will grow best by reason of presence of both nutrients from fertilizer and tillage/aeration due to earthworms.

10. Project Timeline 8/248/319/79/149/219/2810/510/1210/1910/2611/211/911/1611/2311/3012/7 Review of Basic Physiological Material Development of Research Project Background Research of Experimental Topic Presentation of Research Proposal Fabrication of Testing Chambers Initiation of Experimental Design Data Collection Statistical Analysis of Data Writing of Final Paper Presentation of Completed Research

11. Budget Proposed Budget Category Salaries & Wages Professional $25,000 Technician $10,000 Subtotal $35,000 Benefits $5,000 Travel $500 Non-Expendables SPSS Software $900 Expendables Computer $2,000 Supplies & Expenses$2,000 Copying & Telephone$500 Tuition & Fees $10,000 Subtotal $14,500 Total Costs $55,900

12. Works Cited Deibert, E.J. 1997. Soil quality: Impact of conservation tillage. In: 19 th Manitoba-North Dakota Zero Tillage Workshop. January 27-29, 1997. Brandon, Manitoba. Canada. Manitoba-North Dakota Zero Tillage Farmers Association. Pp131-142. Deibert, E.J. 1997. Soil quality: Impact of conservation tillage. In: 19 th Manitoba-North Dakota Zero Tillage Workshop. January 27-29, 1997. Brandon, Manitoba. Canada. Manitoba-North Dakota Zero Tillage Farmers Association. Pp131-142. Dorn, Philip B., and Salanitro, Joseph P. 2000. Temporal ecological assessment of oil contaminated soils before and after bioremediation. Chemosphere 40: 419-426. Dorn, Philip B., and Salanitro, Joseph P. 2000. Temporal ecological assessment of oil contaminated soils before and after bioremediation. Chemosphere 40: 419-426. Eijsackers, H., Beneke, P., Maboeta, M., Louw, J.P.E., and Reinecke, A.J. 2005. The implications of copper fungicide usage in vineyards for earthworm activity and resulting sustainable soil quality. Ecotoxicology and Environmental Safety 62: 99-111. Eijsackers, H., Beneke, P., Maboeta, M., Louw, J.P.E., and Reinecke, A.J. 2005. The implications of copper fungicide usage in vineyards for earthworm activity and resulting sustainable soil quality. Ecotoxicology and Environmental Safety 62: 99-111. Eriksen-Hamel, Nikita S., and Whalen, Joann K. 2006. Impacts of earthworms on soil nutrients and plant growth in soybean and maize agroecosystems. Agriculture, Ecosystems and Environment 120: 442-448. Eriksen-Hamel, Nikita S., and Whalen, Joann K. 2006. Impacts of earthworms on soil nutrients and plant growth in soybean and maize agroecosystems. Agriculture, Ecosystems and Environment 120: 442-448. Gibbs, Mitchell H., Wicker, Linda F., and Stewart, Arthur J. 1996. A method for assessing sublethal effects of contaminants in soils to the earthworm, Eisenia foetida. Environmental Toxicology and Chemistry 15: 360-368. Gibbs, Mitchell H., Wicker, Linda F., and Stewart, Arthur J. 1996. A method for assessing sublethal effects of contaminants in soils to the earthworm, Eisenia foetida. Environmental Toxicology and Chemistry 15: 360-368.