Sporulation on Blood Agar

From 4-7-15.

I made more Columbian sheep's blood agar slants and plates. The agar was made by autoclaving Columbian agar base. After the agar was cool to the touch, a volume of 5% defibrinated sheep's blood was added to the molten agar. A volume of 8 mL was added to sterile glass tubes to create the slants. The tubes were tilted on their sides while cooling to create the slant (used Linda P.'s tube rack in order to achieve the tilt without dumping out the agar).

100 uL of an overnight broth culture of P. larvae 9545 (in BHI) was spread onto the agar plates. The plates were incubated, inverted, at 37C with 5% CO2 for seven days. On Saturday, April 25, I will extract the P. larvae spores from the agar.

300 uL of the overnight P. larvae 9545 broth culture in BHI was inoculated onto the agar slants. An image of how this process is performed and what it should look like is seen here. The slants were incubated at 37C in the lab incubator for ten days. On Tuesday, April 28, I will extract the P. larvae spores from the slants.

//EWW