Basic laboratory and field manual for conducting research with the entomopathogenic nematodes, Steinernema and Heterorhabditis, and their bacterial symbionts

Abstract

Broad spectrum chemical pesticides are harmful to humans and other nontarget organisms. Biological control, which entails the use of natural enemies, is a viable alternative. Isolation, identification, and application of biocontrol agents such as the entomopathogenic nematodes (EPNs), Steinernema and Heterorhabditis and their symbiotic bacteria, Xenorhabdus and Photorhabdus have increased substantially over the last four decades, and the trend continues with advancement of molecular techniques. Yet, there is a need for a simple hands-on guide for their proper identification, classification, and handling, especially for researchers and users who are not totally familiar with these biocontrol agents. Thus, this manual is intended to provide a practical guide for students and researchers interested in or wanting to focus on these organisms. The manual describes the general biology and bionomics of these nematode/bacterium complexes and explains various basic standard protocols and methods frequently used in research and field laboratories ranging from isolation to application methods. Methods for rearing the insects, Galleria mellonella and Tenebrio molitor, which are routinely used in bioassays and recovery of EPNs, are also included.