Add 100cm3 soil into a 1 liter graduated cylinder.
Add tap water to a volume of 1 liter.
Cover cylinder with parafilm, invert to mix, and let settle
for 60 sec.
Decant over 25 mesh screen over 325 mesh screen (use a slow
twirling motion so as not to disturb the soil pellet).
Spray water over 25 mesh screen to extract any needle nematodes,
Longidorus spp., nematodes are collected over 325 mesh screen.
Wash nematodes from 325 mesh screen into a 150 ml beaker
using water.
Pour beaker contents into 50 ml round bottom centrifuge tubes.
Centrifuge tubes at 400xg for 5 min.
Decant supernatant from tubes.
Add sucrose solution (454 g sucrose / liter water) to centrifuge
tubes and mix.
Centrifuge tubes at 400xg for 1 min.
Decant tubes over 325 mesh screen.
Rinse sucrose solution off screen using tap water from underneath
the screen.
Using tap water in a water bottle, rinse nematodes from screen
into a 50 ml beaker (using less than 20 ml water).
Cover beaker with parafilm and store at 4C until nematode
isolation (samples can be stored this way for several weeks). |