Yesterday after being away for about 2 weeks I came back and was able to do a quick inspection and shrink down a couple of colonies.

Again this year, I have one colony with a mix of Amoeba, Nosema and now (this year's new observation) the malpighian tubules stones (or severe irritation/melanization). During my inspection I noticed a dead queen (marked) out in front of this colony. I pulled fresh live bee samples off its cluster and also collected the queen for testing.

1st random bee tested had very severe tubule amoeba with severe Nosema, the 2nd random bee had severe Nosema with infected/stone/melanization of the tubules.

The queen had what looked like amoeba and severely irritated tubules (stones/melanization). The queen also had large cubic crystals all over the place.

My current goal is to find/identify amoeba trophozoites (infectious material after the 21 day cycle that causes infection in other bees). I have an example from an SEM but it is hard to visualize what it would look like on a normal microscope. I am also curious to see what the pre-cyst infection looks like. The irritation, "rash like" on the tubule surface might be it.

https://doi.org/10.1080/0005772X.1926.11095976 (can anyone help me get a copy of this article from 1926?)

I will be getting rid of all the frames and  thoroughly cleaning out the infected boxes.

Note: We haven't had 1st pollen yet and likely another 2 weeks yet. Very slow spring again this year!!
The symptoms are the same as the last 5 years, high mortality, queen failure, most live bees in spring have either Amoeba + Nosema or severe Nosema and cluster disappears quickly once cleansing starts.

The 2 healthy colonies had approximately 1/4-1/2" of dead bees on bottom board, sick colony had 1.25-1.5" of dead bees.

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