Rapid method for recovery of strongylid third stage larvae of parasitic nematodes from small soil samples.
Livestock with access to pasture is generally exposed to infections with parasitic nematode species by uptake of infective third stage larvae (L3) with the grass. L3 can survive on pasture and particularly also in the soil up to several months and sometimes even longer, depending on temperature and humidity. As indicators for health and productivity of grazing animals it is important to determine the intensity and species spectrum of parasitic nematode larvae by analysing grass as well as soil samples. A rapid method for the recovery of L3 using a centrifugal-flotation technique from soil samples of 50-500 g was developed. The method takes advantage of the low specific weight of larvae to separate them from equal sized soil and debris particles by centrifuging them in a saturated sugar solution. A stack of differently sized sieves is used to achieve elimination of larger particles, dust and sugar from the sample to enable easy counting of larvae. Independent of the number of larvae used for inoculation of the samples a mean recovery of 75.3% was obtained. The recovery rates obtained ranged between 60.8% and 88.0% which demonstrates a considerably lower variability compared to earlier approaches and therefore a more precise estimation of the actual numbers of parasite larvae in soil is achieved. Further advantages over already developed methods are the use of easy, affordable and eco-friendly materials, the simplicity of the procedure and a faster processing time with the possibility to examine up to 20 samples per day.
Institute for Parasitology and Tropical Veterinary Medicine, Freie Universität Berlin, Robert-von-Ostertag-Str. 7-13, 14163 Berlin, Germany.
Friederike
F
Knapp-Lawitzke
Institute for Parasitology and Tropical Veterinary Medicine, Freie Universität Berlin, Robert-von-Ostertag-Str. 7-13, 14163 Berlin, Germany.
Georg
G
von Samson-Himmelstjerna
Institute for Parasitology and Tropical Veterinary Medicine, Freie Universität Berlin, Robert-von-Ostertag-Str. 7-13, 14163 Berlin, Germany. Electronic address: j.demeler@fu-berlin.de.
Janina
J
Demeler
D000818:Animals; D002417:Cattle; D002498:Centrifugation; D007814:Larva / Q000145:classification / Q000254:growth & development; D012987:Soil / Q000469:parasitology*; D014251:Trichostrongyloidea / Q000145:classification / Q000254:growth & development / Q000302:isolation & purification*
D012987:Soil
Centrifugal-flotation technique; Infective third stage larvae; Larval recovery; Nematodes; Soil