Thiourea as a polyphenol oxidase inhibitor enhances host infection by the entomopathogenic nematode, Heterorhabditis beicherriana

Entomopathogenic nematodes (EPN) are potent microbial biocontrol agents applied commercially against various economically important insect pests. The efficacy of EPN in the field may be partially reduced when compared to chemical insecticides due to its sensitivity to environmental stressors like temperature, desiccation, and UV light. Thus, researchers have developed methods to enhance the effectiveness of EPN, such as screening or breeding stress-resistant strains and creating adjuvants for EPN protection in the field. This study used another strategy to lower the host's immunity so that the anti-infection ability of the pest wanes, and the virulence of EPN waxes accordingly. In addition to its use as a plant growth regulator, thiourea also functions as an inhibitor of polyphenol oxidase (PO) in insects. Thiourea can inhibit a key interaction in the humoral immunity of insects, so we utilized it to enhance the efficacy of EPN. To investigate the effects of thiourea on PO activity inhibition and EPN pathogenesis, we studied the effects of thiourea on survival, infectivity, reproduction, and host searching ability of the EPN, Heterorhabditis beicherriana, and monitored in vivo and in vitro polyphenol oxidase activity of the model insect host, Galleria mellonella. The survival of H. beicherriana infective juveniles (IJs) was barely affected after exposure to 0.5 mM and 1 mM thiourea solution, and the reproductive output of H. beicherriana IJs (after applying 50, 100 and 200 IJs per host) in 0.5 mM and 1 mM thiourea was not different compared to the control. However, the LD50 of G. mellonella larvae caused by H. beicherriana without thiourea was significantly higher than H. beicherriana IJs in thiourea. Moreover, thiourea inhibited in vivo and in vitro polyphenol oxidase activity of G. mellonella infected by EPNs at different concentrations. Thus, thiourea can improve the biological control efficacy of EPNs, and the enhanced environmentally friendly approach may apply to diverse cropping systems.