Beauveria bassiana exhibits strong virulence against Dendroctonus ponderosae in greenhouse and field experiments.

The mountain pine beetle (MPB) has infested over 16 million hectares of pine forests in western Canada, killing over 50% of mature lodgepole pine, Pinus contorta, in British Columbia alone. There are few tools available to manage irruptive bark beetle populations and to mitigate tree mortality. Beauveria bassiana is an entomopathogenic fungus that causes mortality to several bark beetle species. However, the potential for B. bassiana as a biocontrol agent against pine beetle populations is unknown. We selected three strains of B. bassiana from several culture collections and evaluated their conidial stability under cold storage, in planta (greenhouse, and pine bolts) and in natura (forest stand, pine bolts, and live pines) conditions. The stability assays showed that all fungal strains maintained a minimum effective conidial yield through the assay durations (3-12 weeks). In addition, we adapted a biphasic liquid-solid fermentation approach for the large-scale production of conidial biomass, yielding up to a 100-fold increase in production. In greenhouse virulence assays, the mean lethal time of MPBs was reduced to 3-4 days upon treatment with B. bassiana, where high B. bassiana-associated mycosis was also observed. Furthermore, the application of B. bassiana formulation substantially affected the gallery network of MPBs in bolts in the field, resulting in shorter larval galleries and significantly reduced offspring production. Indeed, high titer treatments reduced the mean larvae per gallery to virtually zero. Together these results demonstrate that B. bassiana may be a viable biocontrol tool to reduce mountain pine beetle populations in pine forests in western Canada. KEY POINTS: • Three B. bassiana strains identified to be stable at various test conditions. • Large-scale conidial biomass production using liquid-solid biphasic fermentation. • Reproductive success of D. ponderosae significantly reduced by B. bassiana formulation.

Selection of entomopathogenic fungus Beauveria bassiana (Deuteromycotina: Hyphomycetes) for the biocontrol of Dendroctonus ponderosae (Coleoptera: Curculionidae, Scolytinae) in Western Canada.

The mountain pine beetle, Dendroctonus ponderosae, has infested over ~16 Mha of pine forests in British Columbia killing >50% of mature lodgepole pine, Pinus contorta, trees in affected stands. At present, it is functionally an invasive species in Alberta, killing and reproducing in evolutionarily naïve populations of lodgepole pine (P. contorta), novel jack pine (P. banksiana), and their hybrids. The entomopathogenic fungus Beauveria bassiana has shown some potential as a biocontrol agent of several bark beetle species. In this study, nine isolates of B. bassiana were examined for insect virulence characteristics, including conidiation rate, pigmentation, and infection rate in laboratory-reared D. ponderosae, to assess for their potential as biocontrol agents. The strains were categorized into three phenotypic groups based on pigmentation, conidial density, and myceliation rate. Virulence screening utilizing insect-based agar medium (D. ponderosae and European honeybee Apis mellifera carcasses) revealed no difference in selection of fungal growth. However, infection studies on D. ponderosae and A. mellifera showed contrasting results. In vivo A. mellifera infection model revealed ~5% mortality, representing the natural death rate of the hive population, whereas laboratory-reared D. ponderosae showed 100% mortality and mycosis. The LT50 (median lethal time 50) ranges from 2 to 5 ± 0.33 days, and LT100 ranges from 4 to 6 ± 0.5 days. We discuss the selective advantages of the three phenotypic groups in terms of virulence, pigmentation, conidial abundance, and tolerance to abiotic factors like UV and host tree monoterpenes. These results can further provide insights into the development of several phenotypically diverse B. bassiana strains in controlling the spread of the invasive D. ponderosae in Western Canada. KEY POINTS: • Three B. bassiana morphotype groups have been demonstrated to kill D. ponderosae. • A range of effective lethal times (LT50 and LT100) was established against D. ponderosae. • Variable tolerance to UV light and pine monoterpenes were observed in B. bassiana.

Nest boxes increase reproductive output for Tree Swallows in a forest grassland matrix in central British Columbia.

Secondary cavity-nesting birds depend on tree cavities for nesting and roosting, but many studies of these birds are conducted using nest boxes. Implementation of effective conservation strategies for cavity-nesting species such as nest-site supplementation requires careful comparisons of fecundity and other vital rates for birds using both natural and artificial nest site types. We compared breeding phenology, clutch and brood sizes, and fledging success of Tree Swallows (Tachycineta bicolor) nesting in tree cavities and nest boxes during 2001-2003 in British Columbia, Canada. Swallows using nest boxes initiated egg-laying and hatched young at approximately the same time as those in tree cavities (2 June, 23 June, respectively). Female Tree Swallows in boxes laid larger clutches (5.9 ± 0.9 eggs, N = 76) than those in tree cavities (4.2 ± 1.6 eggs, N = 67). The mean number of nestlings hatched was greater in nest boxes (5.2 ± 1.1 nestlings, N = 67) than in tree cavities (2.6 ± 2.0 nestlings, N = 58). Pairs in boxes were over twice as successful in producing fledglings (93.4%; 57 of 61 pairs fledged > 1 young) than those in tree cavities (35.8%; 19 of 53 pairs). Of those successful nests, pairs nesting in boxes fledged 5.1 ± 1.1 young (N = 57), whereas those in tree cavities fledged 3.5 ± 1.2 young (N = 18). Because cavities in nest boxes averaged 60% larger in volume and 1.8 cm wider internally than tree cavities, we suggest that increased reproductive output was correlated with boxes enabling a larger clutch size. In previous research, we found that Tree Swallows were a poor competitor with other cavity-nesting passerines for tree cavities. The addition of nest boxes may serve as an effective way to supplement local reproduction for secondary cavity-nesting bird populations by reducing competition for limited nest sites. This is especially true in regions where the availability of natural nesting sites is highly variable, and where species compete with many other cavity-nesting passerines using a similar ecological niche and nesting cavities.