Wax Worm/P. larvae LD50

From 4-17-15.

I attempted to count survival of early instar wax worms that were exposed to the bacterial endospores in the 96 well plate, however it was incredibly difficult to do so for a number of reasons. It appears that a number of the wax worms were able to escape the wells. This was evident by the discovery of several of them crawling around inside the secondary container and several wells that were mysteriously empty. It appears, that despite my attempts to keep them from escaping by flipping the plate upside down and only putting a very small hole in their covers, that they have still managed to escape.

It was also very difficult to determine if a wax worm still in the well was alive or just not moving at the moment. The wax worms were able to move underneath the oat inside their well, further obstructing my view. I would need to flip the 96 well plate back over to its correct orientation and open it up to more thoroughly assess their survival. However, that is not a feasible thing to do given the circumstances. It should be noted that it is very easy to determine if the wax worm is dead sometimes, especially when they have turned black in color - which some of them had.

Overall, here on Day 1, there was a large amount of wax worms that were unaccounted for in their wells, or were too obscured to accurately determine their survival. I will continue to monitor this plate, however my hopes are low while using this particular set-up.

New Set-up

I repeated the same method as before by transferring early instar wax worms to the wells of a 96 well plate containing a single oat soaked with spores, however this time I sealed the plate using a transparent sticky cover slip. I pieced an even smaller hole than last time in the cover over each well. I am not sure if the wax worms will still be able to escape, but it is significantly easier to observe their survival now (you can see through both the top and the bottom of the plate!).

96 well plates containing the early instar wax worms. The transparent cover slips are affixed to the plates.

The plates (x2) were incubated at 37C inside a secondary container. I will monitor their survival every 24 hours.

//EWW