Identification of entomopathogenic nematodes and their symbiotic bacteria in national parks of Thailand, and mosquitocidal activity of Xenorhabdus griffiniae against Aedes aegypti larvae

Abstract

Entomopathogenic nematodes (EPN) Steinernema and Heterorhabditis with symbionts with Xenorhabdus and Photorhabdus bacteria, respectively, are reported as biocontrol agents for insect control. The objectives of this study were to identify EPN and their symbiotic bacteria in national parks of Phitsanulok Province, Thailand, and to test how Xenorhabdus isolates attack Aedes aegypti larvae. We collected 810 soil samples from four national parks. The juvenile stage of EPN was isolated from soil samples using a baiting technique with Galleria mellonella followed by a White trap. Partial regions of 28S rDNA and internal transcript spacer were sequenced to identify EPN, and recA sequencing was used to discriminate between Xenorhabdus and Photorhabdus. We found that 74 of the 810 soil samples (9.1%) were positive for the EPN. The EPN were molecularly identified as S. surkhetense, S. longicaudum, H. indica and Heterorhabditis sp. SGmg3, and their symbiotic bacteria were identified as X. stockiae, X. griffiniae, X. indica, X. vietnamensis, P. luminescens subsp. akhurstii, and P. temperata subsp. temperata. Xenorhabdus griffiniae showed potential larvicidal activity against A. aegypti (91% mortality at 72 and 96 h after exposure). This study demonstrates the diversity of EPN and symbiotic bacteria in national parks of Thailand and the potential to use X. griffiniae as a biocontrol agent to kill A. aegypti larvae.