Mycoremediation via proliferation of native terrestrial fungi

Methods

Summary

- Twenty 1-cup samples are taken from a 40ft² plot of infertile / potentially contaminated soil (10 samples for heavy metals testing; and 10 samples for nutrient composition testing). These samples are to be sent to Cornell University for soil analyses.

- Using sterile techniques, approximately 2 grams of soil from each sample is sprinkled onto individual Petri dishes, for a total of 20 cultures.

- After 2 weeks of growth, the indigenous fungal growth is separated from bacterial growth by isolating the fungi onto new Petri dishes.

- After 2 more weeks of growth, the new cultures are transferred into grain-based substrates. Such as rye berries; rye grass seed; oats; and/or wild birdseed.

- When the grain-based substrates are colonized, they are used to inoculate 10 pound bags of woody / soil-based substrates.

- When the 10 pound bags are fully colonized, they are thoroughly mixed into the 40ft² plot. The soil is watered thoroughly every day.

- The soil is re-sampled after 3 months of growth. (10 samples for heavy metals testing; and 10 samples for nutrient composition testing).

- The initial and final samples are compared, and conclusions are drawn.

Challenges

- Lack of indigenous fungal diversity. Some genera of fungal, such as Trichoderma & Aspergillus, are extremely prolific in soil. Therefore, I may struggle to obtain any basidiomycetes from my samples. Therefore, limiting my remediation to strictly ascomycetes. Perhaps this factor can be overcome by taking my samples from varying depths. For, different species of fungi reside at different depths in soil.

- Non-indigenous contamination. Though unlikely if sterile techniques are used, it is always possible for foreign species (from the air & substrate) to contaminate the Petri dishes. If this occurs my results will be invalidated, based on the fact that I'm not utilizing indigenous species for remediation. To avoid this, I will conduct several trials leading up to the official isolation of indigenous fungi. If 100% of my trials prove that no contamination occurs, I can safely assume that my isolations will be purely indigenous.

- Poor outdoor growth. When reintroducing the indigenous fungi back into the soil in bulk amounts, it's possible that the fungi will struggle to grow. This can occur because of unfavorable environmental factors, such as heat; light exposure; moisture; and competitors. I may have to control heat and light exposure by shading the plot of soil with a tarp. I already know for sure that I will need to water the soil every day. I should be checking temperature and moisture content of the soil every day. And, I'm not worried about competitors because it's safe to assume that my isolations will already be some of the most prolific species in that soil.

Pre Analysis Plan

The data analysis should be very straight-froward. I will have 10 initial data points describing the concentration of heavy metals in the soil; and 10 initial data points describing the concentration of nutrients in the soil. The averages of each set of data will be taken. After the indigenous fungi are proliferated, the soil will be resampled. I will then have 10 final data points describing the concentration of heavy metals in the soil; and 10 final data points describing the concentration of nutrients in the soil. The averages of each set of data will be taken. Finally, the initial and final averages will be compared. These are the potential outcomes: heavy metals decreased in concentration; heavy metals increased in concentration; or heavy metals concentration did not change. And: total nutrient concentration decreased; total nutrient concentration increased; or total nutrient concentration did not change. Concluding one of these results will tell us whether or not the proliferation of indigenous fungi can be used as a practical bioremediation technique.