Analogous to the early days of developing constructed treatment wetlands using aquatic plants, interest in fungal-based biofiltration systems is growing. The concept of utilizing cultivated fungi to perform environmental services in water treatment applications warrants future research to realize its potential as a new ecological engineering technique. While the results of the present study are preliminary, the use of S. rugoso-annulata mycelium as an amendment for improving E. coli removal by wood-based biofiltration media appears promising. To allow for a more complete interpretation of biological monitoring data, future efforts to assess pathogen removal by mycofiltration should include multiple approaches for microbial identification in biofilter outflow, including chromogenic media, plating, and phylogenetic analysis. Thorough analysis of any correlation between E. coli indicator bacteria removal and waterborne pathogen removal will also be essential to fully understand the value of mycofiltration and to support its adoption as a regulated management practice. Future efforts would also be improved by comparing E. coli removal between material produced using sterile and non-sterile methods. While sterile methods are not likely to be applicable or practicable in the field, a comparison between these methods would illuminate possible confounding variables in this study, such as a resident population of coliform bacteria on some substrate types. Additional research is also needed to explore straw as a potential source of thermotolerant coliform bacterial contamination. This research is of particular regulatory significance due to the widespread use of straw as a stormwater treatment practice and the growing number of bacteria total maximum daily load (TMDL) regulatory actions nation-wide.Further investigations using straw from multiple sources could also help determine if straw as a substrate is susceptible to harboring thermotolerant coliform bacteria, as suggested by this study.