pollen mites- pickling solution?

[toomanyhandles]

Hello-

I have cleaned up my mason bee cocoons and I have a volume of pollen mites isolated- probably about 35cc worth if you were to put them in a 50ml tube.

I have looked around a little but I'm not sure what I should pickle them in (ratios) to keep them from shrinking or popping.

I have 95% ethanol, probably some reagent grade glycerol that I have lost track of, and distilled water around.

These will likely be 'hairy fingered' mite- Chaetodactylidae / Chaetodactylus. Not varroas or larger/ tougher animasl.

Any thoughts on the best solution to pickle them in? I have billions and woudl be happy to send samples to anyone who wants to give a try at imaging them. I thought I might be able to find a pdf of
Krantz, G.W. et al (various years) A manual of acarology.
someplace but no luck yet, that might have some mite-specific protocols.

Brian.

[toomanyhandles]

I am thinking this one seems about right.
The pollen mites are mainly mites but they will have pollen and small bits of clay/dirt from the maon bee chamber walls, I'm not set up at home to do various floats or washes to try to separate things out further.

---
Formula for Koenike's fluid:
Glacial acetic acid 10 mL
Distilled water 40 mL
Glycerin, 50 mL
--

Thanks for thoughts!

Brian.

[MichaelG.]

I have zero experience in this field, but was sufficiently interested in your question to investigate a little …

This looks a very useful resource:
https://idtools.org/bee_mite/index.cfm?pageID=1722

MichaelG.

[toomanyhandles]

That's a great link, thank you. I hadn't found that site yet, it has nice summaries.

Here are the mites in question, from that same site:
https://idtools.org/bee_mite/index.cfm? ... ntityID=88

You can see they probably need cleared for the most interesting microscope views, even though these aren't as sclerotized as the "big" ticks we see on us or dogs (Dermacentor, Amblyomma..)

For clearing them, lactophenol or Nesbitt's solution seems the way, each have components that aren't great for home use, that's what a garage is for perhaps. Compared to the the PFAS/PFOS in all our drinking water anyway, a little phenol or whatever vapor- meh.

Just to get the media listed here:
"Nesbitt's fluid is stronger clearing agent (40g chloral hydrate, 25mL distilled water, and 2.5 mL concentrated acetic acid)"

lactophenol:
"Clearing agents for mites are similar to those used for collembolans. A popular one is lactophenol (Krantz, 1978)":
Lactic acid 50 parts
Phenol crystals 25 parts
Distilled water 25 parts

Clearing agent solution details are from this section of the link below
Laboratory and Field Exercises in Soil Ecology
David C. Coleman, ... Paul F. Hendrix, in Fundamentals of Soil Ecology (Second Edition), 2004:
https://www.sciencedirect.com/topics/ea ... -hydroxide

That article also states:
Generally speaking, mite and collembolan specimens should be archived in 70% alcohol, although “constant vigilance” is necessary to guard against evaporation of the preservative (Christiansen, 1990).

which mention of 70% makes sense to me; I think I will put some back in 70% ethanol, and some in Koenike's, since I can just make that up at home, and see what they look like re: distortion.