Recently I have collected eriophyid mites from leaf erineum of a medicinal plant. When I prepare slide in Hoyer's medium, it is not getting cleared fully, instead I could see fine oil globblets on the entire body surface. I could see only the body margin clearly and difficult to study/keyout using surface structure. Out nearly more than 50 specimens, hardly one specimen was fairly OK and I could make out characters upto generic level, with difficulty. It belongs to Aceria genus and further study has not been possible. (Presently we are working on the integrative taxonomy of mites; for mprphological data we are supplementing DNA data using COI gene/marker) GenBank matching in Blast analysis shows maximum matching of 82% with Aceria genus .
Can any one suggest me to get clear eriophyid specimens. The same Hoyer's medium is being used for tetranychid, tarsonemid, phytoseiid and other familly mites, with absolutely no problem and getting very clear specimens. Even with Aceria mangiferae, A. cajani and other common Aceria species this problem of specimen clearing is not noticed.
Pl. do comment on my problem and suggest me corrective measure......
From
Srinivas
UAS, Bangalore, Karnataka, India
Dear Srinivas,
First, I'd give my highest recommendation to the following publication:
Article Recommended procedures and techniques for morphological stud...
The authors, I am sure, would be happy to provide a copy. The references therein are also useful.
From your question above, it seems you are not clearing your specimens prior to mounting - although you should do this for most or all mites. That stage can help remove oils. Eriophyoidologists also use different media than the usual lactic acid/nesbitts for clearing with PVA/Hoyer's for the mounting medium. I think these different media make better slides for eriophyoids, and recommend these too.
Can you see that the mite is oily, or filled with white crystalline substances? If the latter, I have cleared some very tiny specimens in 5% KOH. That's like the heavy artillery on mites though! I sit and watch them clear, and remove them immediately once the white material is dissolved. They then need to be washed in ethanol before mounting.
Regarding your Blast, 82% is pretty low... one must be careful with the interpretation of this. I have seen 80% matches with the wrong class, which can be embarrassing if one doesn't stop and think about the result.
I hope this helps,
Owen.
Dear Srinivas,
The best suggestion has been suggested by Professor Seeman. You should clear the specimens before mounting slides. Keifer's procedure can be difficult for whom it is not an Eriophyidologist, please read that important and the best article regarding this issue (recommended procedures and techniques for morphological studies of Eriophyoidea (Acari: Prostigmata) de lillo et al. 2010).
If you want to prepare slides with Hoyer's medium or Heinz's medium, I have a suggestion for you: put the specimens to Lactic acid for 3 or 7 days in room temperature or Lactophenol (be attention, it is toxic) after that you can mount the slides (you should drown the mites into liquids, if them mites were in the surface the liquids it can be not clear)
Be careful, for identify the Eriophyid mites you should have the female specimens in Dorsal and Lateral position (at least 10 clear specimens) and also if you have nymphs or males, you will can prove the Host-specify.
Kind regards
Arash Honarmand
If you are in India I suggest you to contact with Dr. Samiran Chakrabarti, for more information.
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