Evaluation of Indigenous Entomopathogenic Nematodes as Potential Biocontrol Agents against Popillia japonica (Coleoptera: Scarabaeidae) in Northern Italy

SIMPLE SUMMARY: The Japanese beetle Popillia japonica is considered one of the most harmful organisms in the world for crops and the urban landscape. Popillia japonica spends most of its life cycle in the soil as a larva. At this stage, this beetle is more susceptible to biological control agents like entomopathogenic nematodes, which are obligate parasites of main soil-inhabiting insects, killing their host in just a few days. In 2014, P. japonica was detected in Northern Italy between the Piedmont and Lombardy regions. This research aims to investigate the natural occurrence of indigenous and locally adapted entomopathogenic nematodes along the Piedmont part of the Ticino river and test the most performing of them via bioassays. Natural isolates were recovered from 39 out of the 155 soil samples collected. The virulence of all entomopathogenic nematodes assessed by laboratory and semi-field assays highlighted that two isolates resulted in more efficiency in controlling P. japonica grubs. This result is very encouraging, and the use of these natural biocontrol agents against this pest is a fundamental component of eco-friendly management. ABSTRACT: The natural presence of entomopathogenic nematodes (EPNs) has been investigated in the Piedmont region (Northern Italy) in areas infested by the Japanese beetle Popillia japonica. Thirty-nine out of 155 soil samples (25.2%) were positive for EPNs. Most of the samples contained only steinermatids (92.3%), 5.1% contained heterorhabditids, and one sample (2.6%) contained both genera. All the recovered isolates were identified at species level both morphologically and molecularly. Steinernema carpocapsae was the most abundant and it was mainly distributed in open habitats, such as perennial meadows, uncultivated soils, and cropland, characterized by sandy loam soil texture and acidic pH. Steinernema feltiae has been found associated mainly with closed habitats such as coniferous and deciduous woodland, characterized by sandy loam-texture and extremely acidic soil. The three isolates of Heterorhabditis bacteriophora were collected only in open habitats (perennial meadows and uncultivated fields) characterized by strongly acidic soils with sandy loam texture. The virulence of all EPN natural strains was evaluated by laboratory assays against P. japonica third-instar larvae collected during two different periods of the year (spring, autumn). The results showed that larval mortality was higher for pre-wintering larvae than post-wintering ones. The five more promising EPN isolates were tested in the semi-field assay in which H. bacteriophora natural strains have been shown to be more efficient in controlling P. japonica grubs. All of these results are finally discussed considering the use of these natural EPNs as biological control agents against P. japonica, within an eco-friendly perspective of management.