Honors College Theses

Publication Date



Biology (B.S.B.)

Document Type and Release Option

Thesis (open access)

Faculty Mentor

Dr. Vinoth Sittaramane


In early 2014 Freedom Industries, located in Charleston, WV, leaked approximately 10,000 gallons of 4-methylcyclohexane methanol into the Elk River. This river serviced approximately 300,000 people as a source of municipal water. Its effects on the people and surrounding wildlife is largely unknown.

Chemicals in surface runoff have the potential to induce aquatic animal trait changes, such as altered movement and feeding, that can ultimately alter predator-prey dynamics in aquatic systems. However, chemicals with high potential to enter aquatic systems, like 4-methylcyclohexane methanol (MCHM) used to clean coal, were tested using only short-term, single-species studies prior to use. Data from short-term MCHM exposures to zebrafish (Danio rerio) suggests that three-hour exposures to 1 ppm MCHM yield 50% reduction in zebrafish activity. However, cellular level changes in molecular toxicity is unknown.

The goal of this experiment was to determine how 4-Methylcyclohexanemethanol (MCHM) impacted the expression of hsp70, p450, and ahr2 genes in Danio rerio species. This was done by extracting RNA using Invitrogen TRIzol protocol. From there cDNA was synthesized and analyzed with qPCR to determine level of gene expression.

The results from this experiment show there was an increase in all 3 biomarkers at 0 ppm, 1 ppm, and 5 ppm compared to the housekeeping gene beta-actin. There was also an increase in all 3 biomarkers between week 1 and 3 of exposure compared to the housekeeping gene beta-actin.