Check survival of remaining larva under dissecting microscope. Two of the confirmed alive larva are now dead and two of the questionably alive larva are also dead now. The remaining surviving larva were fed 20 uL of BAD and continued to incubate at 35C in the anaerobic chamber at >90% humidity.
I noticed the pools of BAD that the larva were on were unusually dry, perhaps the humidity needs adjusting? I will continue to monitor survival and feed the remaining larva.
I performed the first step in creating new spore stocks using the spread plates from 7-5-14 that were created from Spore Stocks B and C. Those plates have been incubating at 37C 5%CO2 for 7 days. Plates containing nearly a complete lawn of bacteria. The bacteria appeared on the agar at a time appropriate for P. larvae as well as contained uniform colonies similar to what P. larvae should look like (flat, white, well defined colonies).
15 mL of sterile ddH2O was added to each plate at 5 mL intervals, plate was gently rubbed with a sterile spreader, and the solution was pipetted and transferred to 50 mL conical tubes. These two solutions, named Spore Stocks D (from the stock B plate) and Spore Stock E (from the stock C plate) were stored at 4C for the time being and are awaiting further processing. Spores will be extracted using the modified spore extraction method from 6-24-14.
//EWW
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