The effect of synthetic female sex pheromone on the transmission of the fungus Metarhizium brunneum by male Agriotes obscurus click beetles.

Autodissemination techniques can potentially be used to distribute insecticides, including microbial insecticides, to cryptic pests. This approach is reliant on the target insect either passing the pathogen passively to other insects or the pathogen cycling within the population after the initial host dies. Here we examine, in small scale experiments, whether male Agriotes obscurus click beetles passively transmit the spores of the fungus Metarhizium brunneum directly, or indirectly via the environment, and whether this is influenced by exposure to synthetic female pheromone. We found that the beetles did not avoid M. brunneum spores and that this behaviour was not affected by pheromone. Exposure to pheromone increased beetle movement and uptake of spores, but this did not result in an increase in infected beetles under our conditions. Beetles were able to transfer spores at high levels via environmental contamination. However, contamination of the environment declined rapidly after exposure to the spores. The results are discussed in the context of developing an autodissemination strategy for click beetles.

Biogeography and genotypic diversity of Metarhizium brunneum and Metarhizium robertsii in northwestern North American soils.

The biogeography and genotype diversity of Metarhizium species in northwestern North American soils was examined; 20 ecoregions were sampled, including 58 agricultural and 80 natural habitat subsites, and areas that were glaciated during the Pleistocene epoch. One hundred and twenty-nine isolates of M. brunneum, 26 isolates of M. robertsii, four isolates of M. guizhouense, one isolate of M. flavoviride, and 55 isolates of Beauveria were recovered. Metarhizium and Beauveria species were isolated in diverse ecoregions within the study area, but a trend for increased isolation of Metarhizium species in western regions of the study area was observed. Consistent with this observation, the prevalence of M. brunneum and M. robertsii decreased at higher elevations, and the opposite was true for Beauveria. Both M. brunneum and M. robertsii were more commonly isolated from agricultural and natural habitat subsites, and considerable genotypic diversity was observed in both habitats and within the same subsite. Metarhizium robertsii, but not M. brunneum, was more commonly isolated from nonglaciated locations; however, less diversity and richness was observed for M. brunneum recovered from glaciated versus nonglaciated locations consistent with insular biogeography. The study has implications for microbial control strategies in the region.

Effect of Collection Month, Visible Light, and Air Movement on the Attraction of Male Agriotes obscurus L. (Coleoptera: Elateridae) Click Beetles to Female Sex Pheromone.

Elaterid female sex pheromone, while currently used for monitoring the adult life stage (click beetle), has only recently been explored as a potential management tool. Consequently, there is little understanding of how abiotic and biotic conditions influence the response of click beetles to the pheromone. We examined whether the response of male Agriotes obscurus L. (Coleoptera: Elateridae) beetles to a cellulose-based formulation of female sex pheromone ('pheromone granules') is influenced by air movement, presence of visible light, and month of beetle collection. In addition, we investigated the distance from which beetles were attracted to the pheromone granules. Click beetle response was determined by measuring movement parameters in free-walking arena experiments. The response to pheromone was not affected by the presence or absence of visible light. We found that beetles collected earlier in the season had increased activity and interaction with pheromone under moving air conditions, compared to beetles collected later. When controlling for storage time, we confirmed that individuals collected in May were less active than beetles collected in March and April. In the field, beetles were recaptured from up to 14 m away from a pheromone granule source, with over 50% being recovered within 4.4 h from a distance of 7 m or less. Understanding how abiotic and biotic factors affect pest response to pheromone can lead to more effective and novel uses of pheromone-based management strategies.

Genetic diversity of Metarhizium anisopliae var. anisopliae in southwestern British Columbia.

The abundance and genetic diversity of the entomopathogenic fungus, Metarhizium anisopliae var. anisopliae, in southwestern British Columbia (BC) and southern Alberta was examined. The fungus was found to be widespread in soil throughout southwestern BC, and was recovered from 56% of 85 sample sites. In contrast to southwestern BC, no M. anisopliae isolates were recovered in southern Alberta. An automated fluorescent amplified fragment length polymorphism (AFLP) method was used to examine genetic diversity. In excess of 200 isolates were characterized. The method identified 211 polymorphic amplicons, ranging in size from approximately 92 to 400 base pairs, and it was found to be reproducible with a resolution limit of 86.2% similarity. The AFLP method distinguished Metarhizium from other entomopathogenic fungal genera, and demonstrated considerable genetic diversity (25 genotypes) among the reference strains of M. anisopliae isolates examined (i.e. recovered from various substrates and geographical locations). Although 13 genotypes of M. anisopliae var. anisopliae were recovered from southwestern BC soils, the vast majority of isolates (91%) belonged to one of two closely-related genotypes. Furthermore, these two genotypes predominated in urban, agricultural and forest soils. The reasons for the limited diversity of M. anisopliae var. anisopliae in southwestern BC are uncertain. However, findings of this study are consistent with island biogeography theory, and have significant implications for the development of this fungus for microbial control of pest insects.

Environmental and behavioral constraints on the infection of wireworms by Metarhizium anisopliae.

Environmental and behavioral factors that affect the infection of wireworms [Agriotes obscurus L. (Coleoptera: Elateridae)] by a unique isolate of Metarhizium anisopliae Sorokin (Hypocreales: Clavicipitaceae) were studied. After wireworms were placed in soil containing 10(6) M. anisopliae conidia/g and incubated at 6, 12, or 18 degrees C, significant disease development and wireworm mortality occurred only in those wireworms incubated at 18 degrees C. At this temperature, mortality was found to be dependant on the time exposed to the contaminated soil, and a minimum exposure time of 48 h was required to cause significant levels of mortality. Despite the restrictive effect of cooler temperatures on disease development and mortality, infected wireworms did not choose temperatures that inhibited disease development when given the opportunity to do so in a separate experiment. Finally, wireworms were repelled by M. anisopliae-contaminated soil at a rate that increased with the soil conidia concentration, but the rate of emigration was reduced when a food source was present. The results of this study indicate that factors including temperature, time exposed to M. anisopliae, conidia soil concentration, and food availability will affect mortality rates of wireworms and are likely to affect field performance of M. anisopliae as a biological control.

Spinosad interacts synergistically with the insect pathogen Metarhizium anisopliae against the exotic wireworms Agriotes lineatus and Agriotes obscurus (Coleoptera: Elateridae).

We determined that spinosad interacts synergistically with the biocontrol agent Metarhizium anisopliae (Metch) Sorokin to increase the mortality of two wild-collected wireworm species, Agriotes lineatus (L.), and Agriotes obscurus (L.). Bioassays were performed using a M. anisopliae isolate originally acquired from a local wireworm cadaver. M. anisopliae was applied as a soil drench at 3.3 x 10(2) and 10(4) conidia per gram sand, respectively. Soil drenches also were prepared using a commercial formulation of the actinomycete toxins spinosyn-A and spinosyn-D (common name spinosad) at sublethal doses of 1.5, 3, and 6 ppm active ingredient per gram sand. Combined treatments of spinosad and M. anisopliae were synergistic in causing mortality for all spinosad concentrations. Wireworm feeding activity was reduced after exposure to both spinosad and M. anisopliae and was found to be concentration dependent. The high mortality and reduced rate of wireworm feeding suggest that spinosad and M. anisopliae treatment combinations should be tested in the field.

Operating characteristics of full count and binomial sampling plans for green peach aphid (Hemiptera: Aphididae) in potato.

Counts of green peach aphid, Myzus persicae (Sulzer) (Hemiptera: Aphididae), in potato, Solanum tuberosum L., fields were used to evaluate the performance of the sampling plan from a pest management company. The counts were further used to develop a binomial sampling method, and both full count and binomial plans were evaluated using operating characteristic curves. Taylor's power law provided a good fit of the data (r2 = 0.95), with the relationship between the variance (s2) and mean (m) as ln(s2) = 1.81(+/- 0.02) + 1.55(+/- 0.01) ln(m). A binomial sampling method was developed using the empirical model ln(m) = c + dln(-ln(1 - P(T))), to which the data fit well for tally numbers (T) of 0, 1, 3, 5, 7, and 10. Although T = 3 was considered the most reasonable given its operating characteristics and presumed ease of classification above or below critical densities (i.e., action thresholds) of one and 10 M. persicae per leaf, the full count method is shown to be superior. The mean number of sample sites per field visit by the pest management company was 42 +/- 19, with more than one-half (54%) of the field visits involving sampling 31-50 sample sites, which was acceptable in the context of operating characteristic curves for a critical density of 10 M. persicae per leaf. Based on operating characteristics, actual sample sizes used by the pest management company can be reduced by at least 50%, on average, for a critical density of 10 M. persicae per leaf. For a critical density of one M. persicae per leaf used to avert the spread of potato leaf roll virus, sample sizes from 50 to 100 were considered more suitable.