Apart from diagnosis, I am trying to find a method to collect coccidia from stool samples, but the Faust method doesn't seem to be working, because I cannot isolate a satisfactory number of oocysts. Are you aware of such a procedure?
HI. I attach you a reference that could help you:
Gestal, C., Abollo, E., Pascual, S., 1999. Evaluation of a method for isolation and purification of sporocysts of the cephalopod coccidian parasite Aggregata Frenzel, 1885 (Apicomplexa, Aggregatidae). Iberus 17, 115–121.
Hi.
The isolation method varies according to the kind of coccidia.
I attach you a reference that could help you at below:
Arrowood MJ, Sterling CR. Isolation of Cryptosporidium
oocysts and sporozoites using discontinuous sucrose and
isopycnic percoll gradients. J Parasitol 1987;73(2):314-9.
You can try this protocol:
- dilute stool in tap water until you obtain an homogenous sample.
- filter your sample with progressive mesh sizes, until about 100 µm.
- transfer in tubes and centrifuge; keep the pellet and discard the supernatant, then resuspend in tap water; repeat for 3-4 times in order to wash the pellet.
- When the supernatant is clear enough you resuspend the pellet in saturated NaCl solution (350 g/L).
- Centrifuge again and you should obtain a dark ring at the top of you tube; these are oocysts that float in saturated solution. Collect the ring and transfer in new tubes and dilute 1:10 with tap water.
- Centrifuge again and then the oocysts should precipitate at the bottom of the tube.
- Discard the supernatant and then resuspend in potassium dichromate; at this point you should be able to count coccidia in a Burker chamber (they are easily recognizable).
I use this procedures for chicken coccidia. From 2kg of litter you should get around 1x10^7 oocysts, depending on the type of stool or litter, species, time of infection etc..
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