The lethal and sub-lethal consequences of entomopathogenic nematode infestation and exposure for adult pine weevils, Hylobius abietis (Coleoptera: Curculionidae).

Entomopathogenic nematodes (EPN) frequently kill their host within 1-2 days, and interest in EPN focuses mainly on their lethality. However, insects may take longer to die, or may fail to die despite being infected, but little is known about the effects of EPN infection on insects, other than death. Here we investigate both lethal and sub-lethal effects of infection by two EPN species, Steinernema carpocapsae and Heterorhabditis downesi, on adults of the large pine weevil, Hylobius abietis. Following 12h nematode-weevil contact in peat, S. carpocapsae killed a significantly higher proportion of weevils (87-93%) than H. downesi (43-57%) at all concentrations tested. Less than 10% of weevils were dead within 2 days, and weevils continued to die for up to 10 days after exposure (LT(50) of 3 days or more). In a separate experiment, live weevils dissected 6 days after a 24h exposure to nematodes on filter paper harbored encapsulated and dead nematodes, showing that weevils could defend themselves against infection. Some live weevils also harbored live nematodes 6 days after they had been removed from the nematode infested medium. Feeding by weevils was not affected by infection with, or exposure to, either species of EPN. We discuss these results in relation to the use of EPN in biological control against H. abietis.

Simulated roots and host feeding enhance infection of subterranean insects by the entomopathogenic nematode Steinernema carpocapsae.

Steinernema carpocapsae can be effective against root-feeding insects despite its reputation as a sedentary ambusher. In pot experiments, using twigs as surrogate roots and pine weevil larvae as targets, we tested the hypothesis that roots serve as physical routeways and conduits of feeding-associated stimuli, thus enhancing the success of S. carpocapsae applied at the surface against subterranean hosts. Insect mortality was lowest (25%) in the absence of plant material, increased to 48% when twigs linked nematodes and insects, and further increased to 69% when the insects were allowed feed on the twigs. This is the first experimental support for the root-routeway hypothesis.

Competition and intraguild predation between the braconid parasitoid Bracon hylobii and the entomopathogenic nematode Heterorhabditis downesi, natural enemies of the large pine weevil, Hylobius abietis.

In biological control programmes introduced natural enemies compete with indigenous enemies for hosts and may also engage in intraguild predation when two species competing for the same prey attack and consume one another. The large pine weevil, Hylobius abietis L. (Coleoptera: Curculionidae), is an important pest of coniferous reforestation in Europe. Among its natural enemies, the parasitoid Bracon hylobii Ratz. (Hymenoptera: Braconidae) and entomopathogenic nematodes have potential as biological control agents. Both parasitoid and nematodes target the weevil larvae and, hence, there is potential for competition or intraguild predation.In this study, we examine the interaction of B. hylobii with the nematode Heterorhabditis downesi Stock, Griffin and Burnell (Nematode: Heterorhabditidae), testing the susceptibility of larvae, pupae and adults of B. hylobii to H. downesi and whether female parasitoids discriminate between nematode-infected and uninfected weevils for oviposition. In choice tests, when weevils were exposed to nematodes 1-7 days previously, no B. hylobii oviposited on nematode-infected weevil larvae. Up to 24 h, healthy weevils were twice as likely as nematode-infected ones to be used for oviposition. Bracon hylobii females did not adjust clutch size; nematode-infected hosts were either rejected or the parasitoid laid a full clutch of eggs on them.When nematodes were applied to the parasitoid feeding on weevil larvae, the nematodes parasitized the parasitoid larvae, there was a reduction in cocoon formation and fewer cocoons eclosed. Eclosion rate was not reduced when nematodes were applied to fully formed cocoons, but nearly all of the emerging adults were killed by nematodes.

Establishment, persistence, and introgression of entomopathogenic nematodes in a forest ecosystem.

Entomopathogenic nematodes (EPN) are currently marketed worldwide for use in inundative biological control, where the applied natural enemy population (rather than its offspring) is expected to reduce insect numbers. Unlike classical biological control, in inundative control natural enemy establishment is not crucial in order to achieve pest suppression. Field trials in Irish forestry provided the opportunity to test predictions regarding the establishment of two exotic (Steinernema carpocapsae and Heterorhabditis megidis) and two indigenous (Steinernema feltiae and Heterorhabditis downesi) species. Nematodes were inundatively applied to pine stumps to control populations of pine weevil, Hylobius abietis, on three clearcut sites, and their persistence and spread monitored for up to five years. All species were recovered three years after application but only S. feltiae was recovered in years 4 and 5. Limited horizontal dispersal to 20 cm (but not 100 cm) was observed, but the majority of nematodes were recovered close to the area of application. Steinernema feltiae was also recovered from nearby stumps to which it had not been applied, indicating possible phoretic dispersal by weevils or other stump-associated fauna. EPN were not recovered from stumps outside the treated area, suggesting that such dispersal is quite localized. Two strains of S. feltiae (Irish and exotic) were applied. Amplified fragment length polymorphism (AFLP) analysis on 11 populations isolated from soil four years later showed that all had a much closer affinity to the applied Irish strain, suggesting persistence of this genotype and extinction of the exotic one. Some strains were clustered close together, and this is interpreted in the light of possible population genetic scenarios. The findings from the field study confirm predictions based on background knowledge of the species and demonstrate the importance of medium-term studies, as a 3-year study would have overestimated the risk of establishment of exotic species. Short-term persistence and spread of S. carpocapsae, S. feltiae, and H. downesi was also studied in pine forest mesocosms. Similar trends to field results, such as limited horizontal dispersal, even vertical distribution, and more abundant recovery of S. feltiae than of other species, point to the utility of mesocosm studies as a predictive tool.