Entomopathogenic nematodes have been proposed as biological agents for the control of Drosophila suzukii, an invasive pest of small-stone and soft-skinned fruits. Larvae of the fly are susceptible to Steinernema carpocapsae infection but the reaction of immune defenses of the host are unknown. To determine the immune response, larvae were infected with S. carpocapsae and Xenorhabdus nematophila to evaluate the effector mechanisms of both humoral and cellular processes. The symbiont bacteria presented an inhibitory effect on the phenoloxidase cascade with a low level of melanization. Besides, X. nematophila activated the synthesis of putative antimicrobial peptides on the hemolymph of infected larvae. However, those peptides presented a lower antimicrobial activity compared to hemolymph from larvae infected with non-symbiont bacteria. Xenorhabdus nematophila avoided also the phagocytosis response of hemocytes. During in vitro and in vivo assays, S. carpocapsae was not encapsulated by cells, unless the cuticle was damaged with a lipase-treatment. Hemocyte counts confirmed differentiation of lamellocytes in the early phase of infection despite the unrecognition of the nematodes. Both X. nematophila and S. carpocapsae avoided the cellular defenses of D. suzukii larvae and depressed the humoral response. These results confirmed the potential of entomopathogenic nematodes to control D. suzukii.

Drosophila suzukii is a pest of global concern due to its great impact on several crops. The entomopathogenic nematode Steinernema carpocapsae was highly virulent to the larvae of the fly although some immune mechanisms were triggered along the infection course. Thus, to understand the gene activation profile we performed a comparative transcriptome of D. suzukii larvae infected with S. carpocapsae and Xenorhabdus nematophila to map the differentially expressed genes involved in the defence response. The analysis exposed the induction of genes involved in the humoral response such as the antimicrobial peptides and pattern-recognition receptors while there was a suppression of the cellular defence. Besides, genes involved in melanisation, and clot formation were downregulated hindering the encapsulation response and wound healing. After the infection, larvae were in a stress condition with an enrichment of metabolic and transport functionalities. Concerning the stress response, we observed variations of the heat-shock proteins, detoxification, and peroxidase enzymes. These findings set a genetical comprehensive knowledge of the host-pathogen relation of D. suzukii challenged with S. carpocapsae which could support further comparative studies with entomopathogenic nematodes.

Entomopathogenic nematodes (EPNs), especially Heterorhabditis bacteriophora, are promising candidates for biological control of Popillia japonica larvae. Thus, there is a need to study native strains to ensure biological agents adapted to specific environmental conditions to improve their effectiveness in field applications. This work aimed to test the pathogenicity of five Azorean isolates of H. bacteriophora strains to third‐instar larvae of P. japonica under laboratory and semi‐field conditions. All EPNs were able to quickly infect larvae with LT₅₀'s ranging between 3.66 and 4.54 days in controlled laboratory conditions. The semi‐field experiment consisted of an EPN application in buried tubes containing five larvae each in May and October. After 15 days, larvae were recovered and confirmed for nematode parasitism. In May, Az29, Az148, Az170, and Az171 displayed a mean of 37% infected larvae per tube. In contrast, EPNs showed a lower infection ability during the experiment in October, and only Az148 maintained the number of parasitized larvae, with a proportion of 40% of infectivity. The results suggest that native EPNs, like Az148, could be better adapted to climatic and edaphic conditions to control P. japonica larvae.

Entomopathogenic nematodes (EPNs) are closely associated with Popillia japonica and potentially used as their biological control agents, although field results proved inconsistent and evoked a continual pursuit of native EPNs more adapted to the environment. Therefore, we surveyed the Azorean Archipelago to isolate new strains of Heterorhabditis bacteriophora and to evaluate their virulence against the model organism Galleria mellonella under laboratory conditions. Six strains were obtained from pasture and coastal environments and both nematode and symbiont bacteria were molecularly identified. The bioassays revealed that Az172, Az186, and Az171 presented high virulence across the determination of a lethal dose (LD50) and short exposure time experiments with a comparable performance to Az29. After 72 hours, these virulent strains presented a mean determination of a lethal dose of 11 infective juveniles cm^-2, a lethal time (LT50) of 34 hours, and achieved 40% mortality after an initial exposure time of only 60 minutes. Az170 exhibited an intermediate performance, whereas Az179 and Az180 were classified as low virulent strains. However, both strains presented the highest reproductive potential with means of 1700 infective juveniles/mg of larvae. The bioassays of the native EPNs obtained revealed that these strains hold the potential to be used in biological control initiatives targeting P. japonica because of their high virulence and locally adapted to environmental conditions.

The truffle beetle, Leiodes cinnamomeus , is the most important pest in black truffle ( Tuber melanosporum ) plantations during autumn and winter. Entomopathogenic nematodes (EPNs) are promising biological control agents against L. cinnamomeus , while entomopathogenic fungi (EPF) are also commonly used in Integrated Pest Management (IPM) programmes to enhance the control of insect pests. Our aims were to test the virulence of a cold-adapted commercial strain of the EPF Lecanicillium muscarium Ve6 and two EPN species, Steinernema carpocapsae and Steinernema feltiae , against adults of L. cinnamomeus at 15 °C and 10 °C, and to assess the effect of their combined application. Two susceptibility assays were performed using different treatments: (1) immersion of adults in a fungal suspension and (2) direct soil treatment of the EPF. Mortality of L. cinnamomeus was recorded 7 days post-treatment at 15 °C and 15 days post-treatment at 10 °C. Lecanicillium muscarium did not cause significant mortality of L. cinnamomeus . Steinernema feltiae did not cause significant mortality at 15 °C after 7 days but resulted in a statistically significant low mortality (33.3%) at 10 °C after 15 days compared to the control. Steinernema carpocapsae caused the highest mortality rates (63.3–100%) at both temperatures, suggesting it would be the best option to control adults of L. cinnamomeus in field applications. The addition of L. muscarium , did not enhance pest suppression compared to EPNs alone. To optimise IPM strategies for L. cinnamomeus control, further research is recommended to explore potential synergistic effects of combining EPNs with other cold-adapted EPF.