The Effects of Spore Dilution and Quantity of VOC on the Growth of P. destructans : Rutgers Save the Bats Project

Since many of our past experiments focused on the effect our VOC had on mycelial plugs of P.destructans, in this experiment we wanted to shift our focus on the effect our VOC had on the spores of P.destructans. This experiment used our VOC at various parts per million against a dilution of spores going from 0, -1, -2, -3, and -4. We placed duplicate plates, containing the appropriate dilution of spores, inside a mason jar and inoculated the inside wall of the mason jar with the appropriate ppm of our VOC.

This experiment was placed in the incubator for three weeks at 10˚C with ~30% humidity.

After the three weeks, we removed the plates and the results were as follows:

Spore	Control	0.5 ppm	1 ppm	2.5 ppm	5 ppm
0	Thick Lawn	Thick*	NG	NG	NG
-2	Thick Lawn	NG	Some growth*	NG	NG
-3	Lawn	NG	NG	NG	NG
-4	Spotty Lawn	NG	NG	NG	NG
-5	32 & 34 CFU	19 & 33 CFU*	NG	NG	NG

The results indicate that lower than the previously identified 5 ppm, was active against the P. destructans. The results labeled with an * was not expected. We suspect there was either difficulty with the VOC overcoming the spores, that the VOC dissipated more quickly in those jars, or that there was a human error when inoculating the jars with our VOC. The last suspected reason is most likely due to the very small quantity that needed to be placed in each jar.

Ten petri dishes containing various amounts of spores. — Dilutions of spores from 0 (left) to -4 (right).

Spore	Control	0.5 ppm	1 ppm	2.5 ppm	5 ppm
0	2 plates	2 plates	2 plates	2 plates	2 plates
-1	2 plates	2 plates	2 plates	2 plates	2 plates
-2	2 plates	2 plates	2 plates	2 plates	2 plates
-3	2 plates	2 plates	2 plates	2 plates	2 plates
-4	2 plates	2 plates	2 plates	2 plates	2 plates

Spore dilutions and parts per million of VOC in each mason jar.

Mason jars on shelves in an incubator. — Mason jars placed in the incubator containing the spore plates and proper VOC quantity at 10˚C for three weeks.

An image of a mason jar containing VOC, petri dishes with spores on PDA, para film, and tin foil or mason jar cover. — Set up of the mason jars in the incubator.

Two mason jars containing petri dishes of spores. — Here we can see the growth comparison after three weeks between the highest concentration of spores and no VOC exposure (left) versus the lowest concentration of spores and highest ppm of our VOC (right).

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