Procedure
1. Test the soil moistures (e.g., 3, 4, 5, 6, 8, 10, 12, 14, 16, 18,
20, 22, and 25% w/w).
2. Place 100 IJs in 25 μl of distilled water on the bottom of each
well for each treatment, filled with pre-wetted soil to provide
the desired soil moisture.
3. Place one G. mellonella larva in each well on top of the soil.
4. Cover the plate, seal with a cellophane tape and turn
upside down.
5. Incubate for 72 h.
6. Turn over the plates upside down every 8 h during the exposure period, to monitor the upward movement of larva and to
obtain a consistent distribution of IJs.
7. Recover insects from individual wells after 72 h exposure.
8. Record the number of dead insects.
9. Rinse larva in distilled water.
10. Dissect in 0.5% pepsin solution.
11. Incubate at 37
C for 2 h to digest the tissues of insects.
12. Record the number of IJs per insect host.
13. Maintain wells without IJs as the control to monitor the mortality, if any, of insect larva.
8 Mass Production of Nematodes
Entomopathogenic nematodes are effortlessly cultured either
in vivo or in vitro for laboratory experiments or commercial development. G. mellonella is the insect of choice for in vivo processing
since it is made commercially in large quantities, and the agar
medium is used in vitro production because it is made commercially
in large numbers and in in vitro method, agar medium is used.
Paralleled to in vitro method, in vivo production is labor intensive,
lacks economies of scale, and is costly, and the outcome is highquality nematodes.
8.1 Methods
of Nematode
Production
1. In vivo method.
2. In vitro method.
(a) Liquid medium method.
(b) Solid medium method.
8.1.1 In Vivo Method
Most steinernematids and heterorhabditids can be produced in
G. mellonella larvae or other suitable insect hosts.
Mass Production of Nematodes
103
1. Test the soil moistures (e.g., 3, 4, 5, 6, 8, 10, 12, 14, 16, 18,
20, 22, and 25% w/w).
2. Place 100 IJs in 25 μl of distilled water on the bottom of each
well for each treatment, filled with pre-wetted soil to provide
the desired soil moisture.
3. Place one G. mellonella larva in each well on top of the soil.
4. Cover the plate, seal with a cellophane tape and turn
upside down.
5. Incubate for 72 h.
6. Turn over the plates upside down every 8 h during the exposure period, to monitor the upward movement of larva and to
obtain a consistent distribution of IJs.
7. Recover insects from individual wells after 72 h exposure.
8. Record the number of dead insects.
9. Rinse larva in distilled water.
10. Dissect in 0.5% pepsin solution.
11. Incubate at 37
C for 2 h to digest the tissues of insects.
12. Record the number of IJs per insect host.
13. Maintain wells without IJs as the control to monitor the mortality, if any, of insect larva.
8 Mass Production of Nematodes
Entomopathogenic nematodes are effortlessly cultured either
in vivo or in vitro for laboratory experiments or commercial development. G. mellonella is the insect of choice for in vivo processing
since it is made commercially in large quantities, and the agar
medium is used in vitro production because it is made commercially
in large numbers and in in vitro method, agar medium is used.
Paralleled to in vitro method, in vivo production is labor intensive,
lacks economies of scale, and is costly, and the outcome is highquality nematodes.
8.1 Methods
of Nematode
Production
1. In vivo method.
2. In vitro method.
(a) Liquid medium method.
(b) Solid medium method.
8.1.1 In Vivo Method
Most steinernematids and heterorhabditids can be produced in
G. mellonella larvae or other suitable insect hosts.
Mass Production of Nematodes
103