P. larvae Project Pilot

Added 40 uL of BAD to all honey bee larva in the 24 well plates. It appears that the majority of larva had consumed most if not all the present BAD. Added no additional BAD to the eggs/larva that was in the 96 well plates (they do not look like they are doing well still). All larva is still being incubated at 35C. Two pictures of larva growing in the 24 well plates is shown below:

Larva is growing at a steady pace (size is increasing). 

Checked P. larvae colony forming plates that were inoculated on 6-27-14. Plate counts reveal relatively no additional CFU on any dilution. Lawns of bacteria are not present on any other dilution plates that weren't presently seen. They have been incubating for five days now at 37C 5%CO2. I was able to get colony counts from Stocks A on the 10^-5 plate = there were 32 colonies present. Picture of the serial dilutions are seen below. The plates were discarded today.

This means there are 32x10^5 CFU/mL or 3.2x10^6 CFU/mL. This concurs with the 1.448 10^6 CFU/mL determined from the 10^-4 plate on 6-30-14. 

Spot/Drop plates created on  6-30-14 we observed and colony counts were performed. Spore Stocks A, B, and C are shown below. 10 uL of the 10^-1 to 10^-4 serial dilution were inoculated onto MYPGP agar. They have been incubating for two days. 

The colonies were difficult to count the exact amount. Not all dilution colonies were counted, only those that had identifiable single colonies that could be attempted to be counted (~10 - 40 colonies). Again, each dilution was plated in five replicates. The relative CFU counted, average of the five replicates, and standard deviations are listed below for each of the three Spore Stocks A-C.

Spore Stock	Replicate CFU	Average	Std
A (from 10^-3)	22	22.6	4.219004622
	18
	20
	24
	29
B (from 10^-3)	20	21.2	5.357238094
	29
	16
	17
	24
C (from 10^-2)	70	68	8.366600265
	60
	70
	80
	60

The calculated CFU/mL as per the formula laid out on 6-30-14 is displayed below.

Spore Stock	CFU/mL
A	2.26x10^6
B	2.12x10^6
C	6.8x10^5

These calculated CFU/mL roughly concur with what was calculated for Spore Stocks A and B. 

//EWW