The European truffle beetle Leiodes cinnamomeus is the most important pest in black truffle (Tuber melanosporum) plantations. Current control methods against it are inefficient, so entomopathogenic nematodes (EPNs) could play an important role in their population regulation due to their efficacy against many soil-dwelling insect pests. A survey of EPNs and Pristionchus nematodes was conducted in truffle soils of Spain, considering environmental and physical–chemical soil factors. A total of 164 soil samples were collected from forests, productive plantations and null-low productive plantations, representing three distinct black truffle-growing habitat types. EPNs were isolated from seven soil samples (4.3%); four nematodes were identified as Steinernema feltiae and three as Heterorhabditis bacteriophora. Both species were sampled in three types of soil texture (loam, sandy loam or sandy clay loam), characterized by alkaline pH (7.5 to 8.5) and high organic matter (2.1–11.04%). The presence of these EPNs was influenced by habitat type and organic matter content. Pristionchus nematodes were isolated from truffle soil, around truffle fruit bodies and under the elytra of L. cinnamomeus, with Pristionchus maupasi being the most commonly identified species. No significant associations were found between environmental and soil factors and the occurrence of Pristionchus nematodes. These nematodes were found in alkaline soils (pH 7.75 to 8.7), across all seven sampled soil textures, with variable organic matter content (0.73%–5.92%). The ecological trends and the presence of Pristionchus may affect the occurrence of EPNs and their prospective use as biological control agents against L. cinnamomeus in black truffle plantations.

Steinernematid nematodes form a developmentally arrested infective juvenile (IJ) stage at the second moult, when conditions inside the insect host are no longer suitable for further reproduction. In a liquid culture micro-assay two environmental cues were shown to influence the formation of Steinernema feltiae IJs. High nematode population density induced IJ formation, suggesting the presence of a nematode pheromone. Bacterial food and soluble nutrients acted competitively to reduce the frequency of IJ formation. Frequency of IJ formation was greatest when lst-stage juveniles were subjected to IJ-inducing conditions. The optimum temperature range for the IJ induction response was 25–30°C. These findings suggest that maximal IJ production in large-scale liquid culture will depend on the correct balance of nematode population density and nutrient availability at the peak of egg hatching.

Using a simple infection model the infection interaction of Steinernema feltiae (Site 76 strain) Filipjev and Galleria mellonella L. was investigated under laboratory conditions. The model analyses showed that the infection interaction was consistent with a proportion of the S. feltiae infective stages being infectious and, alone amongst the infective-stage population, being capable of infection. The investigation also showed that the infection behaviour of the infectious stages could be reduced to a slightly overdispersed probability of transmission. The importance of these findings with respect to the quantitative description and explanation of infection are discussed.

The efficacy of two entomopathogenic biocontrol agents, Steinernema feltiae (Filipjev) and Verticillium lecanii (Zimmerman) Viégas (reclassified now as Lecanicillium muscarium (Petch) Zare & Gams), against Thrips palmi Karny was investigated. Assessments of the effect of temperature on the efficacy of S. feltiae indicated that higher mortality of T. palmi was recorded at 20°C compared to either 15 or 25°C, whereas significantly higher T. palmi mortality followed application of L. muscarium at 25°C. Testing the control agents efficacy on three host plants; chrysanthemum, sweet pepper and cucumber, under constant temperature and high humidity conditions produced no significant difference in the level of T. palmi larval mortality on each host plant. Incorporating the chemical insecticide imidacloprid with both biological agents in a combined control strategy increased T. palmi juvenile mortality. The potential role of S. feltiae and L. muscarium within integrated pest management programmes for the control of T. palmi is discussed.

The potential of using the entomopathogenic nematode Steinernema feltiae to control the sweetpotato whitefly Bemisia tabaci (Gennadius) has been established in previous laboratory studies. However, laboratory studies can overestimate the level of control achieved by biocontrol agents in the glasshouse. Glasshouse trials are therefore required to confirm laboratory results before full-scale commercial development is considered. Under both controlled laboratory and glasshouse conditions high mortality of second instar B. tabaci (>90% and >80%, respectively) was recorded after application of S. feltiae. The efficacy of the biocontrol agent at various application rates was also investigated, where halving the rate of S. feltiae application caused no significant reduction in B. tabaci mortality on tomato foliage. Steinernema feltiae has shown much potential for incorporation into integrated pest management strategies for the control of B. tabaci.

The present study describes a novel heat-stable, water-stress-related protein with a molecular mass of 47 kDa (designated Desc47) in the entomopathogenic nematode Steinernema feltiae (IS-6). The protein was accumulated about 10-fold (from 7·84±1·85 to 74·09±4·35% relative content level [RCL]) in dehydrated clumps of infective juveniles (IJs), which had lost 34·4% of their initial water content (from 65·1±1·7% to 42·7±0·72%) in a desiccation-tolerance-inducing treatment (97% relative humidity [RH] for 3 days). The appearance of Desc47 was accompanied by trehalose accumulation (from 300 to 600 mg trehalose/g protein) during the process of inducing the IJs into a quiescent anhydrobiotic state. A second cycle of IJ dehydration did not alter the RCL of Desc47 (79·3% for the first cycle and 73·3% for the second cycle). Desc47 retained its high RCL (69·7%) in rehydrated active IJs for 3 days, reaching 51·2% of its initial RCL only after a week. No homology to other known proteins was found by mass-spectrometry electrospray-ion-trap analysis. However, of the 5 sequences obtained from the protein (ranging from 11 to 21 amino acids), the 21-amino-acid peptide NVASDAVETVGNAAGQAG(D/T)AV showed excellent homology (74% identity in 19 amino acids) to the cold-responsive protein COR14b (g6564861) from Triticum aestivum. In the Caenorhabditis elegans predicted proteome database search, the N21 yielded the first-best identity score (59% identity in 17 amino acids) to the CE-LEA homologue protein (g2353333). In plants, COR and LEA are related proteins, heat-stable, which are expressed in response to both dehydration and cold acclimation. The implication of the involvement of Desc47 and the osmoprotectant trehalose in the desiccation-tolerance mechanisms of S. feltiae is discussed.

Previous studies have indicated that between 60 and 80% of a population of entomopathogenic nematodes do not infect their insect hosts at any one period in time. Two hypotheses explain this behaviour: the first that there is a subpopulation of non-infectious nematodes and the second that the non-infectious group is created by inhibitory cues derived from infected insects. Through an experimental approach with the Galleria mellonella–Steinernema feltiae system we show that both mechanisms operate together. When conditions for infection were optimized, the sum of individual infection behaviours was similar to the number infecting as a population, implying observed infection rates are driven by intrinsic mechanisms. In addition, there was evidence that an infected host released a chemical cue into the environment which inhibited subsequent levels of infection. This degree of inhibition was independent of the number of infecting nematodes. Both these mechanisms are dynamic, so the observed proportion of infectious nematodes depended heavily on the time of exposure. The implications of these findings for both the design of laboratory trials and the use of entomopathogenic nematodes in biological control are discussed.

Many studies of entomopathogenic nematodes (Heterorhabditidae and Steinernematidae) have reported that only a small proportion (typically <40%) of infective stages (dauers), even under apparently ideal conditions, actually infect a host. The ‘phased infectivity hypothesis’ is most frequently invoked to explain this pattern of low infection with entomopathogenic nematodes. It proposes that at a given point in time not all individuals are infectious i.e. infectiousness is delayed in some individuals. We tested experimentally several predictions based on this hypothesis. Specifically, if phased infectivity occurs, we should be able to expose dauers to increasing numbers of potential hosts until dauers no longer infect and still be able to recover viable dauers. These recovered dauers which did not infect should be infectious at some later point in time. However, our results do not support the phased infectivity hypothesis for 3 species of Steinernema: most dauers could be recovered in one sampling round when provided with sufficient suitable hosts. In contrast, Heterorhabditis bacteriophora frequently did not infect all available hosts, and infectious dauers were recovered in subsequent sampling rounds. This result is more consistent with the phased infectivity hypothesis, but further research is needed before we can be more confident in the hypothesis. For all species tested, the number of available hosts influenced population levels of nematode infectivity. This suggests that the infection status of hosts can influence whether a dauer infects. Our results indicate that phased infectivity is not a common phenomenon in entomopathogenic nematode dauers, despite the widespread acceptance of this hypothesis.

Infection experiments were conducted to assess the proportion of Steinernema feltiae (Site 76 strain) Filipjev infective juveniles which penetrated into the test host Galleria mellonella L. over an 8-week period. Using a combined ANOVA and infection model approach, the analyses showed that the proportion of infective juveniles which penetrated into the test hosts changed significantly over time. This change was found to be consistent with a fluctuation in the size of a non-infectious population structure within the infective juvenile pool. These fluctuations in the magnitude of the infectious structure would dynamically alter the number of juveniles available for infection in hosts and so impose the observed change in the proportion of juveniles penetrating into hosts, over the 8-week time-course. The empirical and ecological implications of such a dynamically limited pattern of infection and possible future research into the mechanisms responsible for the non-infectious population structure are discussed.