Mycoremediation Experiment Revision October 23, 2023

The soil refertilization experiment procedure was attempted for a total of three trials. During trial one, various species of Trichoderma were isolated on agar plates. These isolated cultures were then transferred into various substrates for further proliferation. Trial one failed during this part of the procedure, for, unintended soil-bound fungi (such as aspergillus) were unexpectedly found proliferating alongside the Trichoderma. The project aimed to specifically utilize soil-bound Trichoderma, therefore the results were invalided.For trial two, the same procedure as trial one was conducted. Again, various species of Trichoderma were isolated on agar plates. This time, a microscope was utilized in order to determine how many species of Trichoderma were present. Unfortunately, at this point in the procedure, it was realized that a crucial confounding variable could invalidate the results of the experiment: some Trichoderma cultures may be the result of air-borne contamination, as opposed to the intended soil-bound Trichoderma. The only way to eliminate this confounding variable would be to either invest in a professional-grade laminar flow hood, or a Mini PCR machine. Both of these pieces of equipment are extremely expensive, and far beyond the budget of this project.

Therefore, the experiment was attempted a third time. For the third trial, an entirely different procedure was utilized. This time, ten glass mason jars were filled with the infertile soil. Then, the soil was saturated with sterilized water. The intention here was to allow the native soil fungi to proliferate under high moisture conditions. Unfortunately, to my surprise, these jars did not produce any significant populations of fungi after four months... This experiment has been extremely useful in allowing me to identify the complications and confounding variables of fungal-based soil refertilization.

Despite, these difficulties in my experiment, I am not giving up on this project. In fact, I am excited to announce an entirely new approach to my fungal-based soil refertilization efforts… As a professional arborist, I accumulate many hardwood wood chips on my property. These hardwood wood chips are the most ideal substrate for growing blue oyster mushrooms (Pleurotus Ostreatus) and Wine Cap Mushrooms (Stropharia Rugosannulata). Both of these species of fungi are native to my region (Northeast US). And, both of these species are extremely aggressive growers. Meaning, it’s quite easy to proliferate large amounts of these fungi. Why is this relevant to my experiment. Many peer-reviewed studies prove that these fungi release digestive enzymes that decompose matter in the soil. Thus, I can still conduct soil refertilization with fungi that are native to my region. Essentially, I'm doing the same experiment with different fungi.

Another positive note is that the initial soil composition lab results can still be used for this experiment. For, the soil has not undergone any change since the beginning of the experiment. Though the procedure for the experiment will look very different now, the end result will still produce answers to the hypothesis nonetheless. I am excited to move forward with proliferating oyster & wine cap mycelia, in preparation for utilizing these fungi in soil refertilization.

Michael Spooner