Chapter 8
Laboratory Techniques for Entomopathogenic Nematodes
Abstract
Entomopathogenic nematodes are beneficial biocontrol agents of insect pests. Before application, they need
to be identified and characterized. Insect hosts like G. mellonella, T. molitor and S. litura are reared in
laboratory conditions for various EPN studies. Isolation techniques of soil nematodes depend on nematode
type. EPNs from infected cadavers are collected using White Trap Method, their pathogenicity is verified by
Koch’s postulates, and are quantified using nematode counting devices. Nematodes are examined either live
or heat killed for morphological studies. Nuclear rDNA has been a beneficial tool for providing markers
associated in delimitation of EPN at diverse taxonomic levels. Environmental tolerance is studied for
tolerance to heat, cold, pH, moisture, desiccation, and hypoxia, respectively. EPNs are mass produced
either in vivo or in vitro for laboratory experiments or commercial development. The virulence and
bioefficacy of EPNs is evaluated.
Keywords Insect rearing, Sampling, Storage, ITS region, Phylogeny, RAPD , RFLP , Ecological
characterization, Mass production
1 Host Insect Rearing
Insect hosts like G. mellonella, T. molitor, and S. litura are reared in
laboratory conditions for various EPN studies. However,
G. mellonella is the most commonly used host insect in view of its
increased susceptibility to most nematodes, feasibility of rearing,
broad accessibility, and potentiality to provide large yields.
1.1 Rearing
of Galleria mellonella
(Greater Wax Moth)
Phylum:
Arthropoda
Subphylum:
Uniramia
Class:
Insecta
Order:
Lepidoptera
Family:
Pyralidae
(continued)
S. Sivaramakrishnan and M. Razia, Entomopathogenic Nematodes and Their Symbiotic Bacteria, Springer Protocols Handbooks,
https://doi.org/10.1007/978-1-0716-1445-7_8,
© The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature 2021
47
Laboratory Techniques for Entomopathogenic Nematodes
Abstract
Entomopathogenic nematodes are beneficial biocontrol agents of insect pests. Before application, they need
to be identified and characterized. Insect hosts like G. mellonella, T. molitor and S. litura are reared in
laboratory conditions for various EPN studies. Isolation techniques of soil nematodes depend on nematode
type. EPNs from infected cadavers are collected using White Trap Method, their pathogenicity is verified by
Koch’s postulates, and are quantified using nematode counting devices. Nematodes are examined either live
or heat killed for morphological studies. Nuclear rDNA has been a beneficial tool for providing markers
associated in delimitation of EPN at diverse taxonomic levels. Environmental tolerance is studied for
tolerance to heat, cold, pH, moisture, desiccation, and hypoxia, respectively. EPNs are mass produced
either in vivo or in vitro for laboratory experiments or commercial development. The virulence and
bioefficacy of EPNs is evaluated.
Keywords Insect rearing, Sampling, Storage, ITS region, Phylogeny, RAPD , RFLP , Ecological
characterization, Mass production
1 Host Insect Rearing
Insect hosts like G. mellonella, T. molitor, and S. litura are reared in
laboratory conditions for various EPN studies. However,
G. mellonella is the most commonly used host insect in view of its
increased susceptibility to most nematodes, feasibility of rearing,
broad accessibility, and potentiality to provide large yields.
1.1 Rearing
of Galleria mellonella
(Greater Wax Moth)
Phylum:
Arthropoda
Subphylum:
Uniramia
Class:
Insecta
Order:
Lepidoptera
Family:
Pyralidae
(continued)
S. Sivaramakrishnan and M. Razia, Entomopathogenic Nematodes and Their Symbiotic Bacteria, Springer Protocols Handbooks,
https://doi.org/10.1007/978-1-0716-1445-7_8,
© The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature 2021
47