Introductions of some forest invasive alien species result in important economic, environmental, and ecological impacts. One approach used by the Canadian Food Inspection Agency to improve the detection of these species is to collect logs from trees in declining health at high risk sites of introduction and to incubate them to obtain insects, if present. Trichoferus campestris (Faldermann) (Coleoptera: Cerambycidae) adults emerged, and live larvae were extracted, from one of two logs taken from a dying Norway maple, Acer platanoides Linnaeus (Sapindaceae), in Mississauga, Ontario, Canada. In its native range, eastern Asia, this beetle is polyphagous, however, in North America there is no host record despite numerous interceptions of larvae in wood packaging material and captures of adults at various post-entry sites. An examination of the feeding damage caused by T. campestris in that maple suggests this insect is not a primary pest of trees in Ontario.

Environmental DNA (eDNA) metabarcoding has revolutionized biodiversity monitoring and invasive pest biosurveillance programs. The introduction of insect pests considered invasive alien species (IAS) into a non‐native range poses a threat to native plant health. The early detection of IAS can allow for prompt actions by regulating authorities, thereby mitigating their impacts. In the present study, we optimized and validated a fast and cost‐effective eDNA metabarcoding protocol for biosurveillance of IAS and characterization of insect and microorganism diversity. Forty‐eight traps were placed, following the CFIA's annual forest insect trapping survey, at four locations in southern Ontario that are high risk for forest IAS. We collected insects and eDNA samples using Lindgren funnel traps that contained a saturated salt (NaCl) solution in the collection jar. Using cytochrome c oxidase I (COI) as a molecular marker, a modified Illumina protocol effectively identified 2,535 Barcode Index Numbers (BINs). BINs were distributed among 57 Orders and 304 Families, with the vast majority being arthropods. Two IAS (Agrilus planipennis and Lymantria dispar) are regulated by the Canadian Food Inspection Agency (CFIA) as plant health pests, are known to occur in the study area, and were identified through eDNA in collected traps. Similarly, using 16S ribosomal RNA and nuclear ribosomal internal transcribed spacer (ITS), five bacterial and three fungal genera, which contain species of regulatory concern across several Canadian jurisdictions, were recovered from all sampling locations. Our study results reaffirm the effectiveness and importance of integrating eDNA metabarcoding as part of identification protocols in biosurveillance programs.

With the increase in global trade and warming patterns, the movement, introduction, and establishment of non‐native insect species has increased. A rapid and effective early detection biosurveillance program to identify species of concern is needed to reduce future impacts and costs associated with introduced non‐native species. One of the challenges facing insect surveillance trapping methods is the sheer volume of individual specimens in the collections. Although molecular identification methods are improving, they currently have limitations (e.g., destructive processing of specimens) and a protocol addressing these limitations can support regulatory applications that need morphological evidence to corroborate molecular data. The novel protocol presented here uses a metabarcoding approach to amplify environmental DNA from a saturated salt solution trap fluid, which retains trap specimens for downstream morphological identifications. The use of a saturated salt solution to preserve specimens in traps addresses issues with the high evaporation rate of ethanol in traps, and public safety concerns with other fluid preservation options with unattended traps in public settings. Using a metabarcoding approach, a 407‐nucleotide segment of the cytochrome c oxidase subunit 1 (COI) animal barcode region was successfully amplified from Lindgren funnel trap collection fluids. These traps were placed in forested areas to survey for wood‐boring beetles of regulatory concern. Our results displayed successful amplification of target taxa, including the molecular identification of the Japanese Beetle Popillia japonica, a species regulated in Canada. A second species, Anisandrus maiche, recently introduced to North America, was identified in every trap. The genus Lymantria, which contains numerous species of concern to North American woodlands, was also detected. Also, there were six other species identified of interest due to their potential impacts on native and crop flora and fauna. Our results show how this protocol can be used as an efficient method for the surveillance of insects using a trap with a saturated salt solution and eDNA metabarcoding to detect species of regulatory concern.

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Boxwood (Buxus spp.) is one of the most commonly planted shrubs in urban landscapes across North America, and in Europe Buxus species comprise a significant portion of the forest understory. Cydalima perspectalis (colloquially known as the box tree moth) is a defoliator capable of causing mortality-inducing damage on boxwood in both natural and managed settings. Native to Asia, C. perspectalis is now established in both Europe and Canada and has been detected multiple times in the U.S. Here we review the life cycle and potential impact of C. perspectalis in Canada and the U.S. We discuss effective integrated management strategies and provide areas where management and detection could be improved. Rigorous detection systems will be crucial to preventing the establishment and future management of C. perspectalis in North America.

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