The worldwide invasion history of a pest ambrosia beetle inferred using population genomics.

Xylosandrus crassiusculus, a fungus-farming wood borer native to Southeastern Asia, is the most rapidly spreading invasive ambrosia species worldwide. Previous studies focusing on its genetic structure suggested the existence of cryptic genetic variation in this species. Yet, these studies used different genetic markers, focused on different geographical areas and did not include Europe. Our first goal was to determine the worldwide genetic structure of this species based on both mitochondrial and genomic markers. Our second goal was to study X. crassiusculus' invasion history on a global level and identify the origins of the invasion in Europe. We used a COI and RAD sequencing design to characterize 188 and 206 specimens worldwide, building the most comprehensive genetic data set for any ambrosia beetle to date. The results were largely consistent between markers. Two differentiated genetic clusters were invasive, albeit in different regions of the world. The markers were inconsistent only for a few specimens found exclusively in Japan. Mainland USA could have acted as a source for further expansion to Canada and Argentina through stepping stone expansion and bridgehead events. We showed that Europe was only colonized by Cluster II through a complex invasion history including several arrivals from multiple origins in the native area, and possibly including bridgehead from the United States. Our results also suggested that Spain was colonized directly from Italy through intracontinental dispersion. It is unclear whether the mutually exclusive allopatric distribution of the two clusters is due to neutral effects or due to different ecological requirements.

Simple and Efficient Trap for Bark and Ambrosia Beetles (Coleoptera: Curculionidae) to Facilitate Invasive Species Monitoring and Citizen Involvement.

Bark and ambrosia beetles (Coleoptera: Curculionidae: Scolytinae & Platypodinae) are among the most damaging forest pests worldwide, and monitoring is essential to damage prevention. Unfortunately, traps and attractants that are currently used are costly, and agencies rely on limited field personnel for deployment. The situation can be greatly aided by 1) the development of cost-effective trapping techniques, and 2) distribution of the effort through the Citizen Science approach. The goal of this study was to test a simple, effective trap that can be made and deployed by anyone interested in collecting bark and ambrosia beetles. Three trap types made from 2-liter soda bottles and, separately, four attractants were compared. Simple, one-window traps performed comparably at capturing species in traps painted or with multiple windows. A comparison of attractants in two-window traps found that 95% ethanol attracted the highest number of species but that Purell hand sanitizer (70% ethanol) and then Germ-X hand sanitizer (63% ethanol) were also effective. A perforated zip-top plastic bag containing Purell hanging over a trap filled with automobile antifreeze attracted the fewest species and individual specimens. Overall, >4,500 bark and ambrosia beetles, including 30 species were captured, representing a third of the regional species diversity. More than three quarters of the specimens were nonnative, representing nearly half of the known regional exotic species. These results suggest that simple one-window soda bottle traps baited with ethanol-based hand sanitizer will be effective and inexpensive tools for large-scale monitoring of bark and ambrosia beetles.

Effect of chipping on emergence of the redbay ambrosia beetle (Coleoptera: Curculionidae: Scolytinae) and recovery of the laurel wilt pathogen from infested wood chips.

Significant mortality ofredbay trees (Persea borbonia (L.) Spreng.) in the southeastern United States has been caused by Raffaelea lauricola, T.C. Harr., Fraedrich, & Aghayeva (Harrington et al. 2008), a fungal symbiont of the exotic redbay ambrosia beetle, Xyleborus glabratus, Eichhoff (Fraedrich et al. 2008). This pathogen causes laurel wilt, which is an irreversible disease that can kill mature trees within a few weeks in summer. R. lauricola has been shown to be lethal to most native species of Lauraceae and cultivated avocado (Persea americana Mill.) in the southeastern United States. In this study, we examined the survival of X. glabratus and R. lauricola in wood chips made from infested trees by using a standard tree chipper over a 10-wk period. After 2 wk, 14 X. glabratus were recovered from wood chips, whereas 339 X. glabratus emerged from nonchipped bolts. R. lauricola was not found 2 d postchipping from wood chips, indicating that the pathogen is not likely to survive for long inside wood chips. In contrast, R. lauricola persisted in dead, standing redbay trees for 14 mo. With large volumes of wood, the potential for infested logs to be moved between states or across U.S. borders is significant. Results demonstrated that chipping wood from laurel wilt-killed trees can significantly reduce the number of X. glabratus and limit the persistence of R. lauricola, which is important for sanitation strategies aimed at limiting the spread of this disease.

Mycangia of ambrosia beetles host communities of bacteria.

The research field of animal and plant symbioses is advancing from studying interactions between two species to whole communities of associates. High-throughput sequencing of microbial communities supports multiplexed sampling for statistically robust tests of hypotheses about symbiotic associations. We focus on ambrosia beetles, the increasingly damaging insects primarily associated with fungal symbionts, which have also been reported to support bacteria. To analyze the diversity, composition, and specificity of the beetles' prokaryotic associates, we combine global sampling, insect anatomy, 454 sequencing of bacterial rDNA, and multivariate statistics to analyze prokaryotic communities in ambrosia beetle mycangia, organs mostly known for transporting symbiotic fungi. We analyze six beetle species that represent three types of mycangia and include several globally distributed species, some with major economic importance (Dendroctonus frontalis, Xyleborus affinis, Xyleborus bispinatus-ferrugineus, Xyleborus glabratus, Xylosandrus crassiusculus, and Xylosandrus germanus). Ninety-six beetle mycangia yielded 1,546 bacterial phylotypes. Several phylotypes appear to form the core microbiome of the mycangium. Three Mycoplasma (originally thought restricted to vertebrates), two Burkholderiales, and two Pseudomonadales are repeatedly present worldwide in multiple beetle species. However, no bacterial phylotypes were universally present, suggesting that ambrosia beetles are not obligately dependent on bacterial symbionts. The composition of bacterial communities is structured by the host beetle species more than by the locality of origin, which suggests that more bacteria are vertically transmitted than acquired from the environment. The invasive X. glabratus and the globally distributed X. crassiusculus have unique sets of bacteria, different from species native to North America. We conclude that the mycangium hosts in multiple vertically transmitted bacteria such as Mycoplasma, most of which are likely facultative commensals or parasites.

Influence of Temperature and Precipitation Anomaly on the Seasonal Emergence of Invasive Bark Beetles in Subtropical South America.

Several invasive bark beetle species have caused major economic and ecological losses in South America. Accurate predictions of beetle emergence times will make control efforts more efficient and effective. To determine whether bark beetle emergence can be predicted by season, temperature, or precipitation, we analyzed trapping records for three introduced pest species of bark beetles in Uruguay. We used trigonometric functions as seasonal predictors in generalized linear models to account for purely seasonal effects, while testing for effects of temperature and precipitation. Results show that all three beetle species had strong but unique seasonal emergence patterns and responded differently to temperature and precipitation. Cyrtogenius luteus (Blandford) emerged in summer and increased with precipitation but was not affected by temperature. Hylurgus ligniperda (Fabricius) emerged in winter and increased with temperature but was not affected by precipitation. Orthotomicus erosus (Wollaston) had a primary emergence in spring, and a smaller emergence in early summer, but showed no significant relationship with temperature or precipitation. This study shows that the emergence of these bark beetle species in Uruguay is influenced by seasonality more than by temperature and precipitation fluctuations. It also shows how seasonality can be easily incorporated into models to make more accurate predictions about pest population dynamics.

Two new Geosmithia species in G. pallida species complex from bark beetles in eastern USA.

Species of Geosmithia are cosmopolitan but understudied fungi, and most are associated with phloem-feeding bark beetles on various woody hosts. We surveyed 207 bark and ambrosia beetles from 37 species in the eastern USA for associated fungi. The community is dominated by species in the G. pallida species complex (GPSC) and included several Geosmithia isolates that appear to be new to science. The new Geosmithia isolates exhibited the characteristic brownish-colored colonies typical for the G. pallida species complex and were phylogenetically resolved as two genealogically exclusive lineages based on a concatenated multilocus data set based on the internal transcribed spacers (ITS) of the nuc rDNA (ITS1-5.8S-ITS2 = ITS), and the translation elongation factor 1-α (TEF1-α), ß-tubulin (TUB2), and RNA polymerase II second largest subunit (RPB2) genes. Two new Geosmithia species, G. brunnea and G. proliferans, are proposed, and their morphological traits and phylogenetic placements are presented.