P. larvae Project Pilot

Made freezer stocks of overnight broth culture of P. larvae grown in BHI+Thiamine:

           800 uL broth culture + 200 uL 80% glycerol    Stored in -20C in Room 114

Plated 100 uL of overnight broth culture onto MYPGP agar in triplicate. Incubated at 37C 5% CO2. Will use these plates for spores in 6-7 days if they aren't overcrowded with growth. I will determine how much growth is present in 2 days and then potentially dilute out the broth culture and plate that. Literature review indicates that too many colonies on a single plate can actually decrease sporulation.

Performed Gram stain of broth culture. Heat fixed a loop full of broth culture onto slide.
           1. Crystal Violet ~1 minute
           2. Iodine ~30 minutes
           3. Alcohol <10 seconds
           4. Safranin ~30 seconds
Gram stain revealed Gram Positive (almost blue in color) rods.

Helped Laura N. make motility agar (Tryptone + Agar + NaCl) today as well. She may have extra plates for which I can gauge the motility of P. larvae.

Brought a small sample of the Bee Artificial Diet (BAD) that we have been feeding the bees at the USDA. Plated 100 uL of BAD onto Nutrient Agar and incubated at 37C, 35C, and 25C. Stored remaining BAD sample at 4C.

//EWW