Thermophilic Actinomycete Isolates From Local Mushroom Compost, Humboldt Co., CA.

Actinomycetes are a group of gram positive bacteria often found in association with soils. In our experiment we attempted to isolate a thermophilic actinomycete from a fungal compost pile created from the waste products of mushroom cultivation. Samples were taken 6" deep within the compost mound where temperatures ranged from 35-40 degrees Celsius. We cultivated viable samples from the compost extracted. Plated specimens were incubated at 55 degrees Celsius to select for heat adapted species. Organisms displaying typical actinomycete features were examined and transferred to new plates for isolation. Isolations were re-grown in broth in preparation for DNA extraction and PCR. The primers used were F243 & R513 at concentrations of 0.5 microliters. This primer pair was a general set used for identification of actinobacteria. After amplification and visualization through electrophoresis, the specimens was sent to be sequenced. Sequencing results confirmed our isolation of an actinomycete in the genus Streptomyces, however more work was required to obtain species level identification. We are currently using primers for S. coelicolor and S. lividans in the pursuit to classify to the species level.