Answer to Case 755

Answer to the Parasite Case of the Week 755:  microfilariae; permanent staining needed for further identification. 

I'd like to give a giant shout out to the Filariasis Research Reagent Resource Center (FR3) which provided this specimen to me for a course I was teaching - the first annual Mayo Clinic Parasitology Workshop. The live microfilariae were a big hit with my course attendees, so I wanted to share the specimen with all of you as well! This particular sample contained Brugia pahangi (orderable through BEI: NR-48896). 

Thanks to all of you who wrote in to share your diagnostic protocols for the microfilariae. In my laboratory, we screen all Giemsa-stained thick and thin blood films for microfilariae using the 10x objective before going to higher power. However, like Florida Fan, we offer a specific test for microfilariae when suspected clinically. For the microfilariae test, we request that blood be submitted in 3.2% sodium citrate (light blue topped tube) and then perform a Knott's concentration. We no longer examine fresh wet preps of blood due to the concern for infectious diseases (so seeing the live microfilariae in this 'safe' NIH sample was a real treat for us). The Knott's involves the addition of 2% formalin to the blood which lyses the RBCs and make the microfilariae easier to identify. We then centrifuge the specimen and make Giemsa-stained, methanol-fixed smears for examination.

You can read more about the human infecting microfilariae found in blood in a paper I wrote with Blaine Mathison and Marc Couturier HERE. There is a nice algorithm for differentiating the various species. Enjoy!