We simulated a cave-like environment in order to expose our VOC to the P. destructans in a more life-like environment. This was done by placing an inverted small mason jar inside a larger wide mouth mason jar. We then placed a 60mm Petri plate on top of the inverted mason jar. The Petri plate contained a P.destructans mycelial plug placed on PDA. The large mason jar was about 950mL in volume and was covered with a manipulated tin foil. We cut a small sized square shaped hole out of the middle of our tin foil and taped a membrane to the foil to cover the hole. This membrane allows the passage of our VOC without other particles entering as well. Our controls were contained in the same setup, but instead of free standing in the incubator, they were placed in a larger jar sealed with parafilm to prevent the VOC from entering.
The experiment was run for three weeks, keeping the large environmental chamber shut and dark for the entire time. This allowed us to maintain 10˚C and 5 ppm of our VOC over this time frame uninterrupted.
Our results, were as expected. The exposed jars did not have any growth from the mycelial plug, while the controls grew as expected. These plugs were removed and subcultured to grow without the presence of our VOC. After some time the VOC treated P. destrucansshowed as much growth as the control mycelial plugs. This indicated the VOC had a fungistatic effect on the P. destructans.