A simple, robust and semi-automated parasite egg isolation protocol.
Large-scale parasite quantification is required for improving our understanding of the epidemiology and genetics of host-parasite interactions. We describe a protocol that uses a low-density salt solution for flotation and centrifugation of nematode eggs. Subsequently, sucrose flotation and precipitation are used to obtain clear egg preparations. Most traditional quantification protocols such as the McMaster technique are unsuited for the standardized processing of large numbers of samples and the analysis of large amounts of feces per sample. Consequently, they are suited only for small-scale surveys. Our protocol, which can be used to analyze up to 6 g of feces, results in clear egg preparations that are concentrated in wells of a microtiter plate and that are suited for digital recording and automated counting. Starting from a fecal suspension in the first flotation solution to a digital recording requires approximately 40 min per 24 samples.
Mes
Ted H M
TH
Netherlands Institute of Ecology (NIOO-KNAW), Centre for Estuarine and Marine Ecology, POB 140, 4400AC, Yerseke, The Netherlands. t.mes@nioo.knaw.nl
Eysker
Maarten
M
Ploeger
Harm W
HW
D000818:Animals; D000829:Animals, Domestic / Q000469:parasitology*; D002498:Centrifugation; D005243:Feces / Q000469:parasitology*; D009348:Nematoda / Q000166:cytology / Q000302:isolation & purification*; D010063:Ovum / Q000166:cytology*; D010270:Parasite Egg Count / Q000379:methods / Q000662:veterinary*