Erwiniaceae bacteria play defensive and nutritional roles in two widespread ambrosia beetles.

Ambrosia beetles are fungal-growing insects excavating galleries deep inside the wood. Their success as invaders increased scientific interest towards them. However, most studies on their microbiota targeted their fungal associates whereas the role of bacterial associates is understudied. To explore the role of abundant microbial associates, we isolated bacteria from active galleries of two widespread ambrosia beetles, Xylosandrus crassiusculus and X. germanus. These isolates were classified within the Erwiniaceae family and through a phylogenetic analysis including isolates from other insects we showed that they clustered with isolates obtained from ambrosia and bark beetles, including Erwinia typographi. The whole genome analysis of the isolate from active galleries of X. crassiusculus suggested that this bacterium plays both a nutritional role, by providing essential amino acids and enzymes for the hydrolysis of plant biomass, and a defensive role, by producing antibiotics. This defensive role was also tested in vitro against fungi, including mutualists, common associates, and parasites. The bacteria inhibited the growth of some of the common associates and parasites but did not affect mutualists. Our study supported the hypothesis of a mutualist role of Erwiniaceae bacteria in ambrosia beetles and highlighed the importance of bacteria in maintaining the symbiosis of their host with nutritional fungi.

Forest Insect Biosecurity: Processes, Patterns, Predictions, Pitfalls.

The economic and environmental threats posed by non-native forest insects are ever increasing with the continuing globalization of trade and travel; thus, the need for mitigation through effective biosecurity is greater than ever. However, despite decades of research and implementation of preborder, border, and postborder preventative measures, insect invasions continue to occur, with no evidence of saturation, and are even predicted to accelerate. In this article, we review biosecurity measures used to mitigate the arrival, establishment, spread, and impacts of non-native forest insects and possible impediments to the successful implementation of these measures. Biosecurity successes are likely under-recognized because they are difficult to detect and quantify, whereas failures are more evident in the continued establishment of additional non-native species. There are limitations in existing biosecurity systems at global and country scales (for example, inspecting all imports is impossible, no phytosanitary measures are perfect, knownunknowns cannot be regulated against, and noncompliance is an ongoing problem). Biosecurity should be a shared responsibility across countries, governments, stakeholders, and individuals.

Response of native and exotic longhorn beetles to common pheromone components provides partial support for the pheromone-free space hypothesis.

Longhorn beetles are among the most important groups of invasive forest insects worldwide. In parallel, they represent one of the most well-studied insect groups in terms of chemical ecology. Longhorn beetle aggregation-sex pheromones are commonly used as trap lures for specific and generic surveillance programs at points of entry and may play a key role in determining the success or failure of exotic species establishment. An exotic species might be more likely to establish in a novel habitat if it relies on a pheromone channel that is different to that of native species active at the same time of year and day, allowing for unhindered mate location (i.e., pheromone-free space hypothesis). In this study, we first tested the attractiveness of single pheromone components (i.e., racemic 3-hydroxyhexan-2-one, racemic 3-hydroxyoctan-2-one, and syn-2,3-hexanediol), and their binary and tertiary combinations, to native and exotic longhorn beetle species in Canada and Italy. Second, we exploited trap catches to determine their seasonal flight activity. Third, we used pheromone-baited "timer traps" to determine longhorn beetle daily flight activity. The response to single pheromones and their combinations was mostly species specific but the combination of more than one pheromone component allowed catch of multiple species simultaneously in Italy. The response of the exotic species to pheromone components, coupled with results on seasonal and daily flight activity, provided partial support for the pheromone-free space hypothesis. This study aids in the understanding of longhorn beetle chemical ecology and confirms that pheromones can play a key role in longhorn beetle invasions.

Effect of Trap Color on Captures of Bark-and Wood-Boring Beetles (Coleoptera; Buprestidae and Scolytinae) and Associated Predators.

Traps baited with attractive lures are increasingly used at entry-points and surrounding natural areas to intercept exotic wood-boring beetles accidentally introduced via international trade. Several trapping variables can affect the efficacy of this activity, including trap color. In this study, we tested whether species richness and abundance of jewel beetles (Buprestidae), bark and ambrosia beetles (Scolytinae), and their common predators (i.e., checkered beetles, Cleridae) can be modified using trap colors different to those currently used for surveillance of jewel beetles and bark and ambrosia beetles (i.e., green or black). We show that green and black traps are generally efficient, but also that many flower-visiting or dark-metallic colored jewel beetles and certain bark beetles are more attracted by other colors. In addition, we show that checkered beetles have color preferences similar to those of their Scolytinae preys, which limits using trap color to minimize their inadvertent removal. Overall, this study confirmed that understanding the color perception mechanisms in wood-boring beetles can lead to important improvements in trapping techniques and thereby increase the efficacy of surveillance programs.

Maximizing Bark and Ambrosia Beetle (Coleoptera: Curculionidae) Catches in Trapping Surveys for Longhorn and Jewel Beetles.

Bark and ambrosia beetles are commonly moved among continents within timber and fresh wood-packaging materials. Routine visual inspections of imported commodities are often complemented with baited traps set up in natural areas surrounding entry points. Given that these activities can be expensive, trapping protocols that attract multiple species simultaneously are needed. Here we investigated whether trapping protocols commonly used to detect longhorn beetles (Coleoptera: Cerambycidae) and jewel beetles (Coleoptera: Buprestidae) can be exploited also for detecting bark and ambrosia beetles. In factorial experiments conducted in 2016 both in Italy (seminatural and reforested forests) and Canada (mixed forest) we tested the effect of trap color (green vs purple), trap height (understory vs canopy), and attractive blend (hardwood-blend developed for broadleaf-associated wood-boring beetles vs ethanol in Italy; hardwood-blend vs softwood-blend developed for conifer-associated wood-boring beetles, in Canada) separately on bark beetles and ambrosia beetles, as well as on individual bark and ambrosia beetle species. Trap color affected catch of ambrosia beetles more so than bark beetles, with purple traps generally more attractive than green traps. Trap height affected both beetle groups, with understory traps generally performing better than canopy traps. Hardwood-blend and ethanol performed almost equally in attracting ambrosia beetles in Italy, whereas hardwood-blend and softwood-blend were more attractive to broadleaf-associated species and conifer-associated species, respectively, in Canada. In general, we showed that trapping variables suitable for generic surveillance of longhorn and jewel beetles may also be exploited for survey of bark and ambrosia beetles, but trapping protocols must be adjusted depending on the forest type.

Acquisition of fungi from the environment modifies ambrosia beetle mycobiome during invasion.

Microbial symbionts can play critical roles when their host attempts to colonize a new habitat. The lack of symbiont adaptation can in fact hinder the invasion process of their host. This scenario could change if the exotic species are able to acquire microorganisms from the invaded environment. Understanding the ecological factors that influence the take-up of new microorganisms is thus essential to clarify the mechanisms behind biological invasions. In this study, we tested whether different forest habitats influence the structure of the fungal communities associated with ambrosia beetles. We collected individuals of the most widespread exotic (Xylosandrus germanus) and native (Xyleborinus saxesenii) ambrosia beetle species in Europe in several old-growth and restored forests. We characterized the fungal communities associated with both species via metabarcoding. We showed that forest habitat shaped the community of fungi associated with both species, but the effect was stronger for the exotic X. germanus. Our results support the hypothesis that the direct contact with the mycobiome of the invaded environment might lead an exotic species to acquire native fungi. This process is likely favored by the occurrence of a bottleneck effect at the mycobiome level and/or the disruption of the mechanisms sustaining co-evolved insect-fungi symbiosis. Our study contributes to the understanding of the factors affecting insect-microbes interactions, helping to clarify the mechanisms behind biological invasions.

National Trade can Drive Range Expansion of Bark- and Wood-Boring Beetles.

Several native species of bark- and wood-boring beetles (Coleoptera) have expanded their range within their native biogeographic regions in the last years, but the role of human activity in driving this phenomenon has been underinvestigated. Here we analyze 3 yr of trapping records of native bark- and wood-boring beetles (Cerambycidae and Scolytinae) collected at 12 Italian ports and their surrounding forests to help elucidate the human role in the movement of native species within their native biogeographic region. We trapped several species that occurred either inside or outside their native distributional range within Italy. Species richness and abundance of those species found in the ports located within their native range were most strongly associated with the amount of forest cover in the surrounding landscape, suggesting that they could have arrived in the ports from the nearby forests. The abundance of the species found outside their native range was instead most strongly linked to the amount of national imports arriving at the port where trapping occurred, suggesting that they were likely introduced to the ports from other parts of Italy. This study demonstrates that national sea transportation can favor species range expansion within a country, and confirms that the forests that surround ports can serve as a source of species that can be potentially moved with exports.

Bark and Ambrosia Beetles Show Different Invasion Patterns in the USA.

Non-native bark and ambrosia beetles represent a threat to forests worldwide. Their invasion patterns are, however, still unclear. Here we investigated first, if the spread of non-native bark and ambrosia beetles is a gradual or a discontinuous process; second, which are the main correlates of their community structure; third, whether those correlates correspond to those of native species. We used data on species distribution of non-native and native scolytines in the continental 48 USA states. These data were analyzed through a beta-diversity index, partitioned into species richness differences and species replacement, using Mantel correlograms and non-metric multidimensional scaling (NMDS) ordination for identifying spatial patterns, and regression on distance matrices to test the association of climate (temperature, rainfall), forest (cover area, composition), geographical (distance), and human-related (import) variables with ß-diversity components. For both non-native bark and ambrosia beetles, ß-diversity was mainly composed of species richness difference than species replacement. For non-native bark beetles, a discontinuous invasion process composed of long distance jumps or multiple introduction events was apparent. Species richness differences were primarily correlated with differences in import values while temperature was the main correlate of species replacement. For non-native ambrosia beetles, a more continuous invasion process was apparent, with the pool of non-native species arriving in the coastal areas that tended to be filtered as they spread to interior portions of the continental USA. Species richness differences were mainly correlated with differences in rainfall among states, while rainfall and temperature were the main correlates of species replacement. Our study suggests that the different ecology of bark and ambrosia beetles influences their invasion process in new environments. The lower dependency that bark beetles have on climate allowed them to potentially colonize more areas within the USA, while non-native ambrosia beetles, being dependent on rainfall, are typically filtered by the environment.

Two newly introduced tropical bark and ambrosia beetles (Coleoptera: Curculionidae, Scolytinae) damaging figs (Ficus carica) in southern Italy.

In summer 2014, the bark beetle Hypocryphalus scabricollis (Eichhoff) and the ambrosia beetle Xyleborus bispinatus Eichhoff, species new to Italy and Europe, respectively, were found for the first time in south-eastern Sicily (Italy). Large infestations of the two species were recorded in many plantations of common fig (Ficus carica L.) both in 2014 and 2015. Data concerning insect characteristics, taxonomy, and distribution are briefly reported.