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Many animals lay their eggs in clusters. Eggs on the periphery of clusters can be at higher risk of mortality. We asked whether the most commonly occurring clutch sizes in pentatomid bugs could result from geometrical arrangements that maximize the proportion of eggs in the cluster’s interior. Although the most common clutch sizes do not correspond with geometric optimality, stink bugs do tend to lay clusters of eggs in shapes that protect increasing proportions of their offspring as clutch sizes increase. We also considered whether ovariole number, an aspect of reproductive anatomy that may be a fixed trait across many pentatomids, could explain observed distributions of clutch sizes. The most common clutch sizes across many species correspond with multiples of ovariole number. However, there are species with the same number of ovarioles that lay clutches of widely varying size, among which multiples of ovariole number are not over-represented. In pentatomid bugs, reproductive anatomy appears to be more important than egg mass geometry in determining clutch size uniformity. In addition, within this group of animals that has lost most of its variation in ovariole number, clutches with a broad range of shapes and sizes may still be laid.

Candidate biological control agents of invasive insect pests are increasingly being found in new geographic regions as a result of unintentional introductions. However, testing the degree of genetic differentiation among adventive and native-range populations of these agents is rarely done. We used reduced-representation sequencing of genomic DNA to investigate the relationships among laboratory lines of Trissolcus japonicus (Ashmead) (Hymenoptera, Scelionidae), an egg parasitoid and biological control agent of the brown marmorated stink bug, Halyomorpha halys (Stål) (Hemiptera, Pentatomidae). We compared sequences from multiple adventive populations in North America (Canada, USA) and Europe (Switzerland) with populations sourced from part of its native range in China. We found considerably more genetic variation among lines sourced from adventive populations than among those within native populations. In the Pacific Northwest of North America (British Columbia, Canada and Washington State, USA), we found preliminary evidence of three distinct genetic clusters, two of which were highly dissimilar from all other lines we genotyped. In contrast, we found that other adventive lines with close geographic proximity (two from Ontario, Canada, three from Switzerland) had limited genetic variation. These findings provide a basis for testing biological differences among lines that will inform their use as biological control agents, and provide evidence to support a hypothesis of several independent introductions of T. japonicus in western North America from different source areas.

Unintentional introduction of natural enemies has increased in recent years due to a massive rise in global trade and tourism. One such natural enemy is the Asian egg parasitoid Trissolcus japonicus, a promising agent for classical biological control of Halyomorpha halys. In Europe, adventive T. japonicus populations have been detected in Switzerland, Italy and Germany. Host specificity testing demonstrated that its fundamental host range is fairly broad; however, it is unclear whether spatial or temporal refuges reduce parasitism of non-target species in the field. To address this, the realized host range of T. japonicus was assessed over three years by exposing sentinel egg masses of H. halys and 18 non-target species and collecting naturally laid egg masses in Switzerland and Italy. In total, 15 of 18 non-target species were successfully parasitized by T. japonicus in the field, confirming its broad fundamental host range. However, most non-target species were less parasitized by T. japonicus than H. halys, profiting from either partial temporal or spatial refuges from parasitism. Species with an unusual life cycle and the same ecological niche as H. halys, such as Pentatoma rufipes, which was the most parasitized non-target species in both countries, potentially face an increased risk of parasitism. In contrast, beneficial non-target effects may occur for the invasive pest, Nezara viridula, which suffered high non-reproductive mortality induced by T. japonicus. In both cases, life table studies will be needed to determine the impact of non-target parasitism and the potential consequences at the population level.

Understanding competition between scelionid parasitoids that exploit the same host may provide insight into strategies that allow coexistence on a shared resource. Competition studies typically focus on interactions between native and exotic parasitoids that do not share an evolutionary history; however, coevolved parasitoids may be more likely to demonstrate strategies to avoid or exploit a shared resource. We examined intrinsic and extrinsic competition between Asian Trissolcus japonicus (Ashmead) and T. cultratus (Mayr) (Hymenoptera: Scelionidae) associated with Halyomorpha halys (Stål) (Hemiptera: Pentatomidae) that share an evolutionary history. Interspecific interactions were assessed by providing parasitized egg masses to each species at various intervals post‐parasitism, and measuring host acceptance, developmental suitability, and guarding behaviour. Trissolcus japonicus showed high acceptance of parasitized hosts up to 72 h following oviposition by T. cultratus, despite a very poor developmental outcome. In contrast, T. cultratus generally avoided ovipositing in H. halys eggs containing T. japonicus early‐instar larvae but did not avoid parasitizing H. halys that contained eggs and third instar larvae. The adaptive value of this behaviour was supported by developmental outcome: T. cultratus outcompeted T. japonicus eggs but not early‐instar larvae, and a trophic shift occurred wherein T. cultratus developed as a facultative hyperparasitoid on third instar T. japonicus larvae. Trissolcus japonicus guarded egg masses 8–12× longer and displayed more aggressive interactions than T. cultratus, suggesting T. japonicus is the superior extrinsic competitor. Development as a facultative hyperparasitoid provided a competitive niche for Asian T. cultratus and confirms its instrinsic competitive superiority. This also occurs in a biologically distinct European population of T. cultratus, suggesting that facultative hyperparasitism as a competitive strategy is retained in geographically separated populations that have not coevolved with H. halys or T. japonicus., Competition between two co‐evolved scelionid egg parasitoids was evaluated to better understand the competitive strategies that allow these two species to coexist on the same host. Not only was Trissolcus cultratus the superior intrinsic competitor when it was the first species to parasitize an egg mass, but it was also able to develop as a facultative hyperparasitoid on T. japonicus. In contrast, T. japonicus displayed more aggressive interactions and longer guarding times, indicating that it is the superior extrinsic competitor.

Predicting how much of a host or prey population may be attacked by their natural enemies is fundamental to several subfields of applied ecology, particularly biological control of pest organisms. Hosts or prey can occupy refuges from natural enemy attack, but habitat or ecological refuges are challenging or impossible to predict in a laboratory setting - which is often where efficacy and specificity testing of candidate biological control agents is done. Here we explore how intraspecific variation in continuous traits that confers some protection from natural enemy attack - even after the natural enemy has encountered the prey - could provide partial refuges. The size of these trait-based refuges should depend on the relationship between trait values and host/prey susceptibility to natural enemy attack, and on how common different trait values are within a host/prey population. These can be readily estimated in laboratory testing of natural enemy impact on target or non-target prey or hosts as long as sufficient host material is available. We provide a general framework for how intraspecific variation in protective host traits could be integrated into biological control research, specifically with reference to non-target testing as part of classical biological control programs. As a case study, we exposed different host clutch sizes of target (pest) and non-target (native species) stink bug (Hemiptera: Pentatomidae) species to a well-studied exotic biocontrol agent, the egg parasitoid Trissolcus japonicus (Hymenoptera: Scelionidae). Although we observed several behavioural and reproductive responses to variation in host egg mass size by T. japonicus, they did not translate to increases in predicted refuge size (proportion host survival) large enough to change the conclusions of non-target testing. We encourage researchers to investigate intraspecific variation in a wider variety of protective host and prey traits and their consequences for refuge size. This article is protected by copyright. All rights reserved.

Predicting how much of a host or prey population may be attacked by their natural enemies is fundamental to several subfields of applied ecology, particularly biological control of pest organisms. Hosts or prey can occupy refuges from natural enemy attack, but habitat or ecological refuges are challenging or impossible to predict in a laboratory setting – which is often where efficacy and specificity testing of candidate biological control agents is done. Here we explore how intraspecific variation in continuous traits that confers some protection from natural enemy attack – even after the natural enemy has encountered the prey – could provide partial refuges. The size of these trait-based refuges should depend on the relationship between trait values and host/prey susceptibility to natural enemy attack, and on how common different trait values are within a host/prey population. These can be readily estimated in laboratory testing of natural enemy impact on target or non-target prey or hosts as long as sufficient host material is available. We provide a general framework for how intraspecific variation in protective host traits could be integrated into biological control research, specifically with reference to non-target testing as part of classical biological control programs. As a case study, we exposed different host clutch sizes of target (pest) and non-target (native species) stink bug (Hemiptera: Pentatomidae) species to a well-studied exotic biocontrol agent, the egg parasitoid Trissolcus japonicus (Hymenoptera: Scelionidae). Although we observed several behavioural and reproductive responses to variation in host egg mass size by T. japonicus, they did not translate to increases in predicted refuge size (proportion host survival) large enough to change the conclusions of non-target testing. We encourage researchers to investigate intraspecific variation in a wider variety of protective host and prey traits and their consequences for refuge size.

The apple leaf midge, Dasineura mali (Kieffer) (Diptera: Cecidomyiidae), an invasive alien pest established for many years in Nova Scotia, Canada, has invaded Ontario and British Columbia, Canada apple (Malus domestica Borkhausen; Rosaceae) orchards, damaging growing tips of trees. Molecular analysis indicated that Nova Scotia populations are genetically different from Ontario and British Columbia populations. Pheromone trap captures, oviposition on growing apple terminals, and the incidence of third instars indicate three D. mali generations in each province. Platygaster demades Walker (Hymenoptera: Platygastridae), released in Nova Scotia in 1993, parasitised 34% of the third midge generation in that province and was reared from D. mali for the first time in 2016 in the Fraser Valley of British Columbia. Lyrcus nigroaeneus Ashmead (Hymenoptera: Pteromalidae) parasitised up to 21% of D. mali in southwestern Ontario. Synopeas myles (Walker) (Hymenoptera: Platygastridae) was recorded from D. mali for the first time, one specimen in each of Nova Scotia and Ontario, and was the most important parasitoid in British Columbia. Synopeas myles parasitism in Okanagan and Similkameen, British Columbia orchards increased from 0% to a mean of 30% of D. mali larvae from 2014 to 2016. Other minor parasitoids included Platygaster tuberosula Kieffer (Hymenoptera: Platygasteridae) in all three provinces and Aphanogmus vicinus Förster (Hymenoptera: Ceraphronidae) in British Columbia.

The arrival, establishment and pest status of Halyomorpha halys in Europe and non-native countries in Asia have been well-documented, with thorough characterisation of the genetic diversity and occurrence of cytochrome oxidase I (COI) haplotypes in Switzerland, France, Hungary, Italy and Greece. However, a number of gaps exist in terms of the characterisation of the haplotype diversity and occurrence of H. halys along the invasion front that covers eastern Europe, western and central Asia. To contribute towards filling this gap, the COI haplotype diversity and distribution were investigated for H. halys collected in Serbia, Ukraine, Russia, Georgia and Kazakhstan. A total of 646 specimens were analysed and five haplotypes were found (H1, H3, H8, H33 and H80). Haplotype H1 was present in all five countries investigated and was the only haplotype detected amongst > 500 specimens collected from Ukraine, Russia and Georgia. H1 (82%) was the dominant haplotype found in Kazakhstan, alongside H3 (18%). In contrast to the low or no diversity observed in these four countries, Serbia had higher haplotype diversity and was represented by five haplotypes. Although H3 was dominant (47%) in Serbia, H1 was also prevalent (40%); the remaining haplotypes (H8, H33 and H80) were minor contributors (1–11%) to the haplotype composition. The results are discussed in context with other known populations in neighbouring countries and patterns of haplotype diversity indicate the movement of successful invasive populations in Europe to generate secondary invasions along the eastern front of the invasion in Eurasia. Possible scenarios regarding the spread of particular haplotypes in these regions are discussed, along with suggestions for future research to fill existing gaps.

Coenosia attenuata is a member of the tigrina-group of Coenosia (sensu Hennig 1964) and is a capable generalist predator in its larval and adult stages. C. attenuata is common in greenhouses worldwide, however, there are few documented cases of its presence in the wild. Here, we estimated C. attenuata presence in the southeastern USA peach orchards using pan traps. Over two years, a total of 717 specimens were collected from both commercially managed and fungicide-only managed peach orchards. C. attenuata is a known biological control agent in artificial greenhouse settings, but its impact on pest species in the wild is still unknown. For the first time in North America, we document an established wild population of C. attenuata, provide an overview of basic identification, and review potential benefits for biological control.

We report the first detection of Trissolcusjaponicus, an exotic Asian egg parasitoid and the primary candidate for classical biological control of the invasive brown marmorated stink bug, Halyomorphahalys, in Canada. Twenty-eight Trissolcusjaponicus emerged from an H.halys egg mass from a site heavily infested by H.halys in Chilliwack, British Columbia, in 2018. This egg mass was deployed and retrieved as part of ongoing sentinel egg mass surveys for natural enemies of H.halys from 2017–2018 in coastal and interior British Columbia (total of 1,496 egg clusters at 16 sites). The identification of T.japonicus was based on biology (high levels of successful emergence from H.halys eggs), morphology, and mitochondrial DNA sequences. Trissolcusjaponicus was not detected at any other survey sites in 2017–2018; however, three species of indigenous egg parasitoids were found attending or emerging from H.halys egg masses at low levels (T.japonicus, the extent of its establishment in British Columbia, and its ultimate impact on H.halys populations remain to be determined. Nonetheless, the detection of this exotic biological control agent in Canada concurrently with regulatory review of its intentional importation and release is emblematic of the current uncertainty around regulatory control on the movement of biological control agents across borders.

Following introduction of Halyomorpha halys in Europe and North America, a concerted effort has been made to characterize host–parasitoid interactions involving H. halys. Yet, it is unclear whether the reported low parasitism in the field is due to the rejection of H. halys by native parasitoids, or the inability of larvae to develop. To determine whether native and exotic pentatomids are equally exploited by native parasitoids in the introduced range of H. halys, sentinel stink bug (native and exotic) egg masses were exposed in different habitats and the incidence of parasitism was determined by rearing and DNA-based approaches. Parasitism estimates were always lower with rearing compared to the molecular method. Egg masses of both native and exotic host species were equally likely to be attacked under natural conditions, supporting the idea that H. halys represents an evolutionary trap for native parasitoids. Lack of parasitoid emergence from H. halys eggs is probably not due to native parasitoids rejecting this host, but rather due to the inability of larvae to complete development. The frequent parasitization of H. halys by native parasitoids highlights the possibility of interspecific interactions with natural enemies considered for introduction as part of biological control programme (e.g. Trissolcus japonicus). Oviposition in eggs of the unsuitable H. halys host by native parasitoid may also have population-level consequences for both parasitoids (e.g. reduced population size, shifts in parasitoid community composition) and pentatomids (e.g. competition with H. halys, deceased population regulation by parasitoids). These effects should be considered when developing biological control strategy for H. halys.

In blueberry crops, there are multiple pest species, and some of those can be suppressed by natural enemies including parasitoid wasps and predators. Parasitoid wasps occur within the environment often tracking pest species for food resources to complete their lifecycle. These small wasps are also sensitive to agricultural environments including agrichemicals, habitat availability, and climate. We investigated how the structure of parasitoid communities varied between organic and conventional blueberry systems, and how the communities of these parasitoids varied within field spatial scales (forested border vs edge vs interior). With the lower intensity of agricultural interventions occurring in organic systems and forested borders, we predicted more stable parasitoid numbers that would be insulated from predicted climate variability. In our study, parasitoids were observed in low abundance in each cropping system, with community structure dependent on both management practice and field position. Unmanaged blueberry fields and forested field borders contained more parasitoid families, and in conventional systems, we saw fewer families present in the field interior as compared to field borders. In this first study to characterize Southern parasitoid communities in blueberry production systems, we observed over 50 genera of parasitoids, with a few dominant families (Braconidae and Ichneumonidae) that would contribute to biological control in blueberry systems. Overall, we captured few parasitoids, which indicates a potential vulnerability in biological control, and the need for further research using other sampling techniques to better understand these parasitoid communities.

The establishment of invasive Halyomorpha halys (Stal) outside of its native range may impact native species assemblages, including other pentatomids and their scelionid parasitoids. This has generated interest in defining species diversity and host-parasitoid associations in this system to better understand the impact of invasive alien species on trophic interactions in invaded regions. Information on scelionid-pentatomid associations in natural habitats is lacking, and species-level identification of these associations can be tenuous using rearing and dissection techniques. Naturally occurring pentatomid eggs were collected in areas where H. halys has established in Canada and were analysed using a modified DNA barcoding approach to define species-level trophic interactions. Identification was possible for >90% of egg masses. Eleven pentatomid and five scelionid species were identified, and trophic links were established. Approximately 70% of egg masses were parasitized; parasitism and parasitoid species composition were described for each species. Telenomus podisi Ashmead was the dominant parasitoid and was detected in all host species. Trissolcus euschisti Ashmead was detected in several host species, but was significantly more prevalent in Chinavia hilaris (Say) and Brochymena quadripustulata (Fabricius). Trissolcus brochymenae Ashmead and Tr. thyantae Ashmead were recorded sporadically. Parasitism of H. halys was 55%, and this species was significantly less likely to be parasitized than native pentatomids. The scelionid species composition of H. halys consisted of Te. podisi, Tr. euschisti and Tr. thyantae. Although these species cannot develop in fresh H. halys eggs, we demonstrate that parasitoids attempt to exploit this host under field conditions.

Climatic requirements of natural enemies have important implications for predicting the outcome of classical biological control programs. A five year study (2011–2015) investigated the ectoparasitoid community of the oilseed rape pest Ceutorhynchus obstrictus (Coleoptera: Curculionidae), in different climate zones across Europe to better understand the distribution, abundance, phenology and thermal requirements of Trichomalus perfectus and Mesopolobus morys (Hymenoptera: Pteromalidae). Data generated were then used to develop bioclimatic models to predict the suitability and potential distribution of each species if introduced into North America for biological control of C. obstrictus. Findings indicated that, although T. perfectus and M. morys occupy the same feeding niche and overlap temporally, there are clear geographic differences in abundance. Trichomalus perfectus appears to be a ‘northern’ maritime species, whereas M. morys is likely a ‘southern’, continental species. If introduced, it was predicted that both species would establish in canola growing regions of North America, although climatic conditions appear to be less favourable for T. perfectus than M. morys. This study showed that variation in regional distributions may occur and these are likely based on differing tolerance to climatic conditions. Understanding these tolerances can enhance precision of ecological niche models that predict potential distributions of parasitoids for introduction as candidate biological control agents. Furthermore, the work reported here provides a basis for future research to determine if climate change may favour species such as M. morys that are adapted to warmer summer conditions.

The pepper weevil,Anthonomus eugeniiCano (Coleoptera: Curculionidae), is the most important pest of pepper (CapsicumLinnaeus; Solanaceae) crops in North America. Native to Mexico, the southern United States of America, and Central America, it is intercepted in Canada when peppers are imported to supplement domestic production. Given the proximity of greenhouse and field production to packing facilities, this pest poses a serious risk to the cultivation of peppers in Canada. Once established, it is difficult to control because immature stages of the weevil are protected within the pepper fruit. As such, chemical control targeting these life stages is not effective, and other strategies, including biological control, may prove useful. To explore the potential for biological control options to manage the pepper weevil in areas at risk in Canada, natural enemy surveys were conducted in southern Ontario following the reports of transient, localised field populations in 2016. Parasitoids belonging to three Hymenoptera families including Pteromalidae (Jaliscoa hunteriCrawford,Pteromalus anthonomiAshmead), Eupelmidae (Eupelmus pulchricepsCameron), and Braconidae (NealiolusMason species,BraconFabricius species) were reared from infested field-collected pepper fruits. Together, these new natural enemy records could facilitate the exploration and development of novel agents for the biological control of the pepper weevil.

We report the detection and adventive establishment of the samurai wasp, Trissolcus japonicus (Ashmead) (Hymenoptera: Scelionidae), a candidate classical biological control agent for the invasive Halyomorpha halys (Stål) (Hemiptera: Pentatomidae), in southwestern Ontario, Canada. Nine naturally laid, field-collected pentatomid egg masses were collected in 2017 and 2018, and two of these egg masses, belonging to Halyomorpha halys and Podisus maculiventris (Say) (Hemiptera: Pentatomidae), produced adults of T. japonicus. The identification of T. japonicus was confirmed using DNA barcoding as well as morphological characteristics. The remaining egg masses yielded adults of the native Telenomus podisi Ashmead (Hymenoptera: Platygastridae) and Anastatus reduvii (Ashmead) (Hymenoptera: Eupelmidae).

The brown marmorated stink bug, Halyomorpha halys (Stål), native to China, Japan, and Korea, has emerged as a harmful invasive pest of a variety of crops in North America and Europe. The Asian egg parasitoid Trissolcus japonicus has been identified as the most promising agent for classical biological control of invasive H. halys populations. A 4-year study evaluated the fundamental and ecological host ranges of T. japonicus as well as its phenology and impact on H. halys populations in fruit orchards in its native range in northern China. In laboratory no-choice tests, developmental suitability of eight non-target host species for T. japonicus was demonstrated by the successful production of progeny on the majority (>85%) of non-target host species tested. In field-collected, naturally laid egg masses, T. japonicus was the most abundant parasitoid associated with H. halys and Dolycoris baccarum, but was also sporadically found in Plautia crossota. Furthermore, it was regularly reared from sentinel egg masses of Menida violacea, Arma chinensis, and Carbula eoa. The only species that did not support development in the laboratory and field was Cappaea tibialis. Besides the benefit of having a high impact on H. halys populations in Northern China, the risk assessment conducted in the area of origin indicates that native Pentatomidae in North America and Europe could be negatively impacted by T. japonicus. Whether the benefits of T. japonicus outweigh the possible risks will have to be evaluated based on the outcome of additional host range studies in the two invaded regions. Electronic supplementary material The online version of this article (doi:10.1007/s10340-017-0863-y) contains supplementary material, which is available to authorized users.

Introduction of exotic natural enemies for biological control of invasive pests may disrupt existing ecological interactions, which may influence the outcome of biological control introductions. The interactions between Asian egg parasitoids, proposed as classical biological control agents of the highly polyphagous invasive pest Halyomorpha halys (Stal), and parasitoids native to the introduced area are largely unknown. Therefore, adult and larval interspecific competition between the exotic Trissolcus japonicus (Ashmead) and the European Anastatus bifasciatus (Geoffroy) was assessed (1) by observing aggressive interactions between adults of the two species following parasitization and (2) by providing each parasitoid species with previously parasitized H. halys egg masses at various time intervals. The results suggest that T. japonicus and A. bifasciatus engage in counterbalance competition, with the former being a superior extrinsic competitor (egg guarding and aggressiveness) and the latter being a superior intrinsic competitor (successful development from multiparasitized eggs of all ages). We suggest that the presence of T. japonicus is unlikely to have a negative impact on A. bifasciatus, and that those two species can coexist and potentially act synergistically in the biological control of H. halys.

Globally, Anastatus species (Hymenoptera: Eupelmidae) are associated with the invasive agricultural pest Halyomorpha halys (Stal) (Hemiptera: Pentatomidae). In Europe, the polyphagous Anastatus bifasciatus (Geoffroy) is the most prevalent native egg parasitoid on H. halys eggs and is currently being tested as a candidate for augmentative biological control. Anastatus bifasciatus frequently displays behavior without oviposition, and induces additional host mortality through oviposition damage and host feeding that is not measured with offspring emergence. This exacerbates accurate assessment of parasitism and host impact, which is crucial for efficacy evaluation as well as for pre- and post-release risk assessment. To address this, a general Anastatus primer set amplifying a 318-bp fragment within the barcoding region of the cytochrome oxidase I (COI) gene was developed. When challenged with DNA of three Anastatus species -A. bifasciatus, Anastatus japonicus Ashmead, and Anastatus sp.-, five scelionid parasitoid species that might be encountered in the same host environments and 11 pentatomid host species, only Anastatus DNA was successfully amplified. When applied to eggs of the target host, H. halys, and an exemplary non-target host, Dendrolimus pini L. (Lepidoptera: Lasiocampidae), subjected to host feeding, no Anastatus amplicons were produced. Eggs of the two host species containing A. bifasciatus parasitoid stages, from 1-h-old eggs to pupae, and emerged eggs yielded Anastatus fragments. Confirmation of parasitoid presence with dissections and subsequent PCRs with the developed primer pair resulted in 95% success for 1-h-old parasitoid eggs. For both host species, field-exposed sentinel emerged eggs stored dry for 6 months, 100% of the specimens produced Anastatus amplicons. This DNA-based screening method can be used in combination with conventional methods to better interpret host-parasitoid and parasitoid-parasitoid interactions. It will help address ecological questions related to an environmentally friendly approach for the control of H. halys in invaded areas.

An adventive aphid and novel host-parasitoid association from cultivated strawberry (Fragaria × ananessa Duch. cv. Chandler; Fragaria × ananessa Duch. cv. Camarosa) in Mississippi, USA are reported herein. The aphid, first detected in high tunnel cultivation, was found predominately on newly emerged, not fully developed leaflets of daughter plants in the Fall of 2016. By 2017, aphids and their associated mummies were observed on fully developed leaflets on mother plants of both cultivars. The aphid was identified as Aphis ruborum (Borner & Schilder) using morphology and DNA barcoding studies. In addition, DNA barcoding identified parasitoid adults emerging from aphid mummies as two cryptic species, Aphelinus varipes (Foerster) and Aphelinus albipodus Hayat and Fatima. Occurrence of A. ruborum in Mississippi represents a new state record and the eastern-most established record in the United States. The A. ruborum - A. varipes or A. albipodus host-parasitoid association is reported for the first time anywhere in the world.

A species of Idris Förster (Hymenoptera: Scelionidae) is found to parasitize the eggs of Bagrada hilaris (Hemiptera: Pentatomidae) and is described as new: Idris elba Talamas, sp. nov. This is the first association of an Idris species with a non-spider host, and the association is confirmed with molecular diagnostic tools that enable identification of parasitoid and host from the remains of parasitized eggs.

The bertha armyworm (BAW), Mamestra configurata, is a significant pest of canola (Brassica napus L. and B. rapa L.) in western North America that undergoes cyclical outbreaks every 6-8 years. During peak outbreaks millions of dollars are spent on insecticidal control and, even with control efforts, subsequent damage can result in losses worth millions of dollars. Despite the importance of this pest insect, information is lacking on the dispersal ability of BAW and the genetic variation of populations from across its geographic range which may underlie potential differences in their susceptibility to insecticides or pathogens. Here, we examined the genetic diversity of BAW populations during an outbreak across its geographic range in western North America. First, mitochondrial cytochrome oxidase 1 (CO1) barcode sequences were used to confirm species identification of insects captured in a network of pheromone traps across the range, followed by haplotype analyses. We then sequenced the BAW genome and used double-digest restriction site associated DNA sequencing, mapped to the genome, to identify 1000s of single nucleotide polymorphisms (SNP) markers. CO1 haplotype analysis identified 9 haplotypes distributed across 28 sample locations and three laboratory-reared colonies. Analysis of genotypic data from both the CO1 and SNP markers revealed little population structure across BAW's vast range. The CO1 haplotype pattern showed a star-like phylogeny which is often associated with species whose population abundance and range has recently expanded and combined with pheromone trap data, indicates the outbreak may have originated from a single focal point in central Saskatchewan. The relatively recent introduction of canola and rapid expansion of the canola growing region across western North America, combined with the cyclical outbreaks of BAW caused by precipitous population crashes, has likely selected for a genetically homogenous BAW population adapted to this crop.

Halyomorpha halys (Hemiptera: Pentatomidae) is an emerging pest which established in Ontario, Canada, in 2012. Halyomporpha halys overwinters in anthropogenic structures as an adult. We investigated seasonal variation in the cold tolerance, water balance, and energetics of H. halys in southwestern Ontario. We also induced diapause in laboratory-reared animals with short daylength at permissive temperatures and compared cold tolerance, water balance, energetics, and metabolism and gas exchange between diapausing and non-diapausing individuals. Halyomorpha halys that overwintered outside in Ontario all died, but most of those that overwintered in sheltered habitats survived. We confirm that overwintering H. halys are chill-susceptible. Over winter, Ontario H. halys depressed their supercooling point to c. −15.4 °C, and 50% survived a 1 h exposure to −17.5 °C. They reduce water loss rates over winter, and do not appear to significantly consume lipid or carbohydrate reserves to a level that might cause starvation. Overall, it appears that H. halys is dependent on built structures and other buffered microhabitats to successfully overwinter in Ontario. Laboratory-reared diapausing H. halys have lower supercooling points than their non-diapausing counterparts, but LT50 is not enhanced by diapause induction. Diapausing H. halys survive desiccating conditions for 3–4 times longer than those not in diapause, through decreases in both respiratory and cuticular water loss. Diapausing H. halys do not appear to accumulate any more lipid or carbohydrate than those not in diapause, but do have lower metabolic rates, and are slightly more likely to exhibit discontinuous gas exchange.

Studies of realized niche shift and model transferability in alien species usually ignore the potential effects of source populations and different invaded-range environments on niche lability. We incorporate our detailed knowledge of the native-range source populations and global introduction history of brown marmorated stink bug (Halyomorpha halys) to examine intraspecific variation in realized niche expansion and unfilling, and to investigate how niche modelling approaches are affected by that variation. Realized niche dynamics of H. halys were analyzed using an ordination method, ecological niche models (ENMs), and occurrence records from (1) East Asia, (2) North America, (3) Europe, (4) native-range source populations for North America and Europe introductions, and (5) global range. Patterns of niche filling and expansion were observed across the invaded ranges of H. halys in North America and Europe: niche unfilling (42.7 %) and expansion (0.0 %) in North America, and unfilling (80.5 %) and expansion (28.0 %) in Europe. Some invasive populations have expanded into climatically novel areas in central Europe. Results presented here provide evidence that H. halys has not yet occupied all suitable climatic habitats in North America and Europe that resemble its native range, and the central USA and most parts of Europe appear to be most at risk of H. halys spread in near future. When realized niche shifts dominated by niche unfilling, fully capturing species’ requirement by basing ENMs on native range may be more important for accurate invasion forecasts than non-native models. Caution is warranted when using the source population to estimate invasion potential.

Hidden trophic interactions are important in understanding food web ecology and evaluating the ecological risks and benefits associated with the introduction of exotic natural enemies in classical biological control programs. Although non-target risk is typically evaluated based on evidence of successful parasitism, parasitoid-induced host mortality not resulting in visible evidence of parasitism (i.e., nonreproductive effects) is often overlooked. The adventive establishment of Trissolcus japonicus, an exotic parasitoid of the introduced stink bug Halyomorpha halys, provides an opportunity to investigate the total impact of this parasitoid on target and non-target hosts in the field. We developed a new methodology to measure nonreproductive effects in this system, involving a species-specific diagnostic PCR assay for T. japonicus. We applied this methodology to field-deployed eggs of four pentatomid species, coupled with traditional rearing techniques. Nonreproductive effects were responsible for the mortality of an additional 5.6% of H. halys eggs due to T. japonicus, and were even more substantial in some of the non-target species (5.4&ndash, 43.2%). The observed hidden mortality of native non-target species from an introduced parasitoid could change predictions about direct and indirect ecological interactions and the efficacy of biological control of the target pest.

Invasive species can destabilize ecological communities by becoming novel hosts, prey, predators or competitors for native species. They can have lasting effects on population dynamics when they reduce the fitness of native individuals. The establishment and spread of invasive Halyomorpha halys, in Europe and North America, have presented native natural enemies in the introduced areas with such a challenge. The native parasitoids readily parasitize eggs of H. halys, but their progeny rarely develops in fresh eggs of this host. The barriers to successful development of native parasitoids in H. halys eggs remain unidentified, and the affected developmental processes therein remain unclear. To determine the timing of failed development of native parasitoids in fresh H. halys eggs, we examined the temporal development of the Trissolcus euschisti parasitoid within suitable (Podisus maculiventis) and unsuitable (Halyomorpha halys) host eggs using a DNA barcode-based approach, and in situ 3D visualization by X-ray micro-computed tomography (micro-CT). Trissolcus euschisi parasitoid development in fresh H. halys eggs fails soon after parasitization either during the egg stage or during early-stage larval development, as limited or no larval development was observed 24 h after initial parasitization. In light of these findings, we propose a time window for further investigation of several potential mechanisms that might result in failed parasitoid development in H. halys.

Foraging parasitoid females should preferentially oviposit on hosts most suitable for progeny development to maximize their fitness. However, the introduction of a new host species may disrupt the link between the reliability of the cues and the expected adaptive outcome of female choice, leading to an evolutionary trap. This mismatch between behavioural acceptance and lack of development exists for North American and European egg parasitoids (Scelionidae) that encounter invasive Halyomorpha halys in areas where this exotic host has recently established. To explain this mismatch, we utilized an L9 orthogonal array design to assess and rank the influence of several critical factors characterizing host resource (host species, egg age, egg status, and surface wash) on behaviour (acceptance, patch residence and patch exploitation) and development of North American native Trissolcus euschisti egg parasitoid. Our results indicate that the host egg age is most important for behaviour, but is least influential for development of the progeny. This study suggests that the maladaptive decision to oviposit in an unsuitable host is due to a mismatch between the cues that females use, and the subsequent expected outcome of this choice. Therefore, it is the relative importance of individual factors when assessed simultaneously that influences the decision-making of female parasitoids.

Drosophila suzukii Matsumura is a recent invasive fruit pest in North and South America and Europe, causing pre-harvest damage and significant economic losses to crops. Little is known about biological control of D. suzukii ; here we report the predatory capability of Dalotia (Atheta) coriaria Kraatz, a commercially available control agent against small dipteran pests, and develop molecular tools to assess predation. Da. coriaria showed a Type II functional response to all D. suzukii instars, with maximal consumption of 26 first, 15 second, and 6 third instars per beetle per day. No pupae were consumed. When raspberries with 11–21 larvae (2nd–3rd instar) per berry were exposed to six Da. coriaria for 4.5 d, a 50% reduction in infestation occurred. A primer pair for D. suzukii was designed that produced a 112 bp amplicon from the mtDNA CO1 gene. Of 19 Drosophila species tested, DNA from one species in the invaded range of D. suzukii was amplified, but it was distinguishable from D. suzukii due to different amplicon melting temperatures. A standard curve was created, relating amount of second instar D. suzukii DNA to Cq values (amount of fluorescence) generated by qPCR. In a feeding trial, the half-life of D. suzukii DNA was 2.3 h in Da. coriaria , but the estimated median detection time was 25.9 h. D. suzukii DNA was detected in Da. coriaria exposed to infested raspberries, but DNA quantity in beetles was only weakly related to predation rates. The control potential of Da. coriaria against D. suzukii should be further assessed, and molecular tools used to identify other D. suzukii predators.

Drosophila suzukii (Matsumura) (Diptera: Drosophilidae) is a worldwide emerging pest of soft fruits, but its cold tolerance has not been thoroughly explored. We determined the cold tolerance strategy, low temperature thermal limits, and plasticity of cold tolerance in both male and female adult D. suzukii. We reared flies under common conditions (long days, 21°C; control) and induced plasticity by rapid cold-hardening (RCH, 1h at 0°C followed by 1h recovery), cold acclimation (CA, 5 days at 6°C) or acclimation under fluctuating temperatures (FA). D. suzukii had supercooling points (SCPs) between -16 and -23°C, and were chill-susceptible. 80% of control flies were killed after 1h at -7.2°C (males) or -7.5°C (females); CA and FA improved survival of this temperature in both sexes, but RCH did not. 80% of control flies were killed after 70 h (male) or 92 h (female) at 0°C, and FA shifted this to 112 h (males) and 165 h (females). FA flies entered chill coma (CTmin) at approximately -1.7°C, which was ca. 0.5°C colder than control flies; RCH and CA increased the CTmin compared to controls. Control and RCH flies exposed to 0°C for 8h took 30-40 min to recover movement, but this was reduced to10 min in CA and FA. Flies placed outside in a field cage in London, Ontario, were all killed by a transient cold snap in December. We conclude that adult phenotypic plasticity is not sufficient to allow D. suzukii to overwinter in temperate habitats, and suggest that flies could overwinter in association with built structures, or that there may be additional cold tolerance imparted by developmental plasticity.

Originating in East Asia, the brown marmorated stink bug, Halyomorpha halys, has established in North America and Europe where it has become an urban and agricultural pest. Although initial populations in Europe were only reported in Switzerland, H. halys has expanded its range to include areas in France, Greece, Hungary, and Italy. Analysis of a portion of the mitochondrial COI gene was used to evaluate the genetic diversity of these populations and to investigate the movement and spread of different COI haplotypes of this pest in Europe. A total of 9 haplotypes are now confirmed in Europe. Four of these haplotypes have not been previously reported elsewhere, and one of these haplotypes has not been previously reported in Europe but is known from Korea. The H. halys population in Greece was the most diverse, with 7 of the 9 haplotypes occurring in the area of collection. Haplotype H1, which is known from North America, Italy, and China, was also present in samples collected in France, Greece, Hungary, and Switzerland. In fact, the H1 haplotype was the dominant haplotype detected in Budapest, Hungary and also occurred frequently in the samples from Athens, Greece. Patterns of haplotype diversity indicate the movement of successful invasive populations to generate secondary invasions within Europe (‘bridgehead effect’), as well as the occurrence of multiple invasions from Asia. Scenarios regarding the movement and spread of particular haplotypes in Europe are discussed.

Halyomorpha halys was accidentally introduced into Switzerland around 2004 and has recently spread to several additional areas in Europe, with established populations documented in France, Italy, Greece and Hungary. To test whether generalist egg parasitoids of European Pentatomidae have the potential to control H. halys, and whether these could be as effective as their related Asian species, sentinel egg masses of H. halys and six native European pentatomids were exposed in two regions of Switzerland. Freeze-killed sentinel egg masses of H. halys were attacked by the platygastrid Trissolcus cultratus and the eupelmid Anastatus bifasciatus, whereas fresh egg masses of native Pentatomidae occassionally yielded four Scelionidae species: Trissolcus semistriatus, T. scutellaris, T. cultratus and Telenomus chloropus. In no-choice tests, T. chloropus and T. semistratus produced no offspring from fresh H. halys eggs, whereas T. cultratus and T. scutellaris were sporadically able to complete development. Stopping embryogenesis by freezing egg masses of H. halys at -80 °C enabled T. cultratus and T. semistriatus to produce a high number of offspring without influencing the behaviour of the parasitoids. In contrast to European platygastrids, the Asian parasitoids Trissolcus japonicus and T. cultratus (Chinese strain) caused significantly higher parasitism of fresh H. halys eggs. Differences in developmental success may be related to less efficient venom of European scelionids injected during oviposition. To date, A. bifasciatus is the only potential candidate for augmentaive biological control of H. halys in Europe as it was the only European parasitoid capable of consistent, successful development on fresh H. halys eggs.

Drosophila suzukii is an emerging global pest of soft fruit; although it likely overwinters as an adult, larval cold tolerance is important both for determining performance during spring and autumn, and for the development of temperature-based control methods aimed at larvae. We examined the low temperature biology of third instar feeding and wandering larvae in and out of food. We induced phenotypic plasticity of thermal biology by rearing under short days and fluctuating temperatures (5.5-19 degrees C). Rearing under fluctuating temperatures led to much slower development (42.1 days egg-adult) compared to control conditions (constant 21.5 degrees C; 15.7 days), and yielded larger adults of both sexes. D. suzukii larvae were chill-susceptible, being killed by low temperatures not associated with freezing, and freezing survival was not improved when ice formation was inoculated externally via food or silver iodide. Feeding larvae were more cold tolerant than wandering larvae, especially after rearing under fluctuating temperatures, and rearing under fluctuating temperatures improved survival of prolonged cold (0 degrees C) to beyond 72 h in both larval stages. There was no evidence that acute cold tolerance could be improved by rapid cold hardening. We conclude that D. suzukii has the capacity to develop at low temperatures under fluctuating temperatures, but that they have limited cold tolerance. However, phenotypic plasticity of prolonged cold tolerance must be taken into account when developing low temperature treatments for sanitation of this species. Crown Copyright (C) 2016 Published by Elsevier Ltd. All rights reserved.

The invasive stink bug Halyomorpha halys Stål (Hemiptera: Pentatomidae) has recently become established in Canada, but little information is available regarding the ability of natural enemies to attack this pest in its invaded range. We tested the capacity of several life stages of three generalist predators, Chrysoperla carnea (Stephens) (Neuroptera: Chrysopidae), Coleomegilla maculata De Geer (Coleoptera: Coccinellidae), and Podisus maculiventris (Say) (Hemiptera: Pentatomidae), to attack H. halys eggs. The relative susceptibility of H. halys eggs to predator attack was compared to P. maculiventris eggs. We found evidence for differential ability to consume H. halys and P. maculiventris eggs both among predator species and within species among life stages. The most H. halys eggs were consumed by late-instar C. carnea, while fewest were consumed by second-instar and adult C. maculata, and second-instar C. carnea. The susceptibility of H. halys eggs to predators was similar to that of P. maculiventris eggs, with only early-instar P. maculiventris nymphs consuming significantly less H. halys eggs than P. maculiventris eggs. Our results provide a baseline for further testing of generalist predators against H. halys and illustrate the potential importance of considering the breadth of ontogeny across which candidates are able to attack the invasive pest.

Halyomorpha halys Stål (Hemiptera: Pentatomidae), the brown marmorated stink bug (BMSB), has been intercepted numerous times in Canada, and overwintering BMSB adults have been reported by homeowners. However, established breeding populations were not detected before 2012. Here we report the occurrence of established populations of BMSB in southern Ontario, Canada and describe habitats, host plants, and collection efforts from 2012 to 2013. Tracking the phenology and spread of this pest in Ontario is critical for the development of integrated pest management strategies to prevent severe economic damage due to this pest.

The invasive Halyomorpha halys was accidentally introduced into Switzerland around 2004 and has recently established in the neighbouring countries of France and Italy. To better understand the population dynamics of this pest in Europe, the phenology, reproductive biology and temperature requirements of Swiss H. halys populations were investigated. Overwintered adults became active in April, but peak oviposition was not observed before early July. Individual females laid on average 79 eggs (maximum of 160). The oviposition period lasted from mid-June to end of September. Eggs laid in August and September did not result in offspring due to the low temperatures in autumn. Under natural fluctuating temperatures, development from egg to adult lasted between 60 and 131 days. The first new generation of adults did not occur before mid-August when the photoperiod was already below 15 h, which likely initiated diapause and suppressed the reproductive activity of new generation adults. Under controlled conditions of 20, 25 and 30 °C, Swiss H. halys populations developed within 75.8, 42.3 and 33.2 days from egg to adult, respectively. No development was possible at or below 15 and at or above 35 °C. The number of degree days required for completion of development from egg to adult was 588.24 DD. Under semi-natural conditions, total mortality of Swiss H. halys populations was 86.7 % with a net reproductive rate of 5.69, indicating growing populations. In Switzerland, H. halys is univoltine, but if it continues to spread into the Mediterranean area two generations per year could be expected.

Invasive alien species can act as ‘evolutionary traps’ for indigenous parasites and predators when the alien species is accepted as prey or a host but is unsuitable for consumption or development. We tested the relationship between acceptance and suitability of eggs of the invasive alien Halyomorpha halys (Hemiptera: Pentatomidae) in North America relative to eggs of the indigenous Podisus maculiventris (Pentatomidae) for the indigenous generalist egg parasitoid Telenomus podisi (Hymenoptera: Scelionidae). T. podisi accepted 0–24 h old H. halys eggs at a rate similar to P. maculiventris eggs (87.5 ± 6.0 and 70.2 ± 9.1 %, respectively). Successful development of T. podisi occurred in 98.3 % of attacked P. maculiventris eggs, but was not observed in H. halys eggs. Oviposition by T. podisi did, however, reduce the developmental success of H. halys embryos relative to unattacked controls by 24.1 % in 0–24 h old eggs and 29.6 % in 24–48 h old eggs. We suggest that as H. halys spreads and increases in abundance in North America, it could operate as an evolutionary trap for indigenous egg parasitoids, thereby indirectly causing an increase in population levels of indigenous pentatomids. This predicted indirect effect would be a result of H. halys eggs acting as an egg sink for T. podisi. We also introduce the concept of a ‘time sink’, which may be particularly relevant for parasitoids such as T. podisi that spend considerable time protecting their reproductive investments.

Halyomorpha halys has recently established in countries outside of its Asian area of origin. In the North Eastern USA, this pest causes severe economic loss in agricultural crops. Breeding populations have been confirmed in Switzerland and Canada; however, their pathways of entry and genetic diversity remain unknown. To determine invasion pathways and source population(s), the diversity of specimens from Asia, North America, and Europe was investigated. Genetic diversity in Asia was higher than that in the Canada and Switzerland. Although three haplotypes were detected among Canadian H. halys, one haplotype dominated. In contrast, two out of three haplotypes were found frequently among Swiss samples. Based on molecular data and interception records it appears likely that H. halys in Canada is derived from the movement of established US populations. Further, North American populations likely originated from the Hebei/Beijing regions of China. The area of origin of the Swiss samples remains unclear. Although the dominant haplotype in Switzerland was consistent with Asian samples collected in the Hebei and Beijing provinces, it was not the dominant haplotype in these regions and further sampling is necessary to its confirm the distribution and abundance in the area of origin. The remaining two haplotypes were unique to Switzerland and no matches were found among our Asian samples. Interestingly, no haplotypes were shared between the North American and Swiss samples, indicating that the two invasions were separate and distinct events and did not result from the movement of goods and materials between USA and Europe.

Genotyping of Ixodes scapularis Say (Acari: Ixodidae) ticks could enhance understanding of the occurrence and genotypes of I. scapularis-borne pathogens. We investigated the utility of mitochondrial (mt) Cytochrome C Oxidase subunit I gene (cox1) sequences as a tool for understanding the population structure of I. scapularis collected in Canada, where we also investigated the geographic occurrence of different cox1 haplotypes. Sequences obtained from 414 ticks were one of 55 unique haplotypes, most of which grouped into one of six clades. Demographic analysis suggested that cox1 sequences have haplotype and nucleotide diversity comparable to other mt genes. All haplotypes were connected in a single minimum spanning network tree. Despite low fixation index values there were significant differences in the frequency of occurrence of haplotypes of different clades among four geographic regions: 1) Alberta to western Ontario, 2) eastern Ontario, 3) Quebec, and 4) Atlantic Provinces; suggesting that cox1 sequences could reveal population structure differences between I. scapularis in geographically separated populations of northeastern and midwestern North America. Spatial clusters of ticks of the same haplotype identified in regions of southern Quebec and southern Ontario where I. scapularis is invading were consistent with population bottlenecks associated with founder events. These findings suggest that cox1 sequences are useful for the study of I. scapularis population structure, are of sufficient diversity that spatial analyses of haplotypes can be used to identify where I. scapularis is emerging in southern Canada, and may be useful for exploring differences between northeastern and midwestern populations of I. scapularis.

The introduction of an exotic species may alter food webs within the ecosystem and significantly affect the biodiversity of indigenous species at different trophic levels. It has been postulated that recent introduction of the brown marmorated stinkbug (Halyomorpha halys (Stål)) represents an evolutionary trap for native parasitoids, as they accept H. halys egg masses as a host but produce no viable progeny. Interspecific interactions between European egg parasitoid, Trissolcus cultratus (Mayr), and an Asian parasitoid, Trissolcus japonicus (Ashmead), were assessed by providing egg masses to T. cultratus at various time intervals following the initial parasitization by T. japonicus. The suitability of the host for the parasitoid development was re‐assessed by providing T. cultratus with fresh and frozen egg masses of various ages. The likelihood of T. cultratus being able to attack previously parasitized egg masses was determined by assessing the duration of egg mass guarding behavior by T. japonicus following parasitization. The results of experiments examining the interspecific interactions between a native European egg parasitoid, T. cultratus, and an Asian parasitoid, T. japonicus (a candidate for the biological control of H. halys), showed that the native species can act as facultative hyperparasitoid of the exotic one. Although this is only possible during certain stages of T. japonicus development, the presence of the introduced parasitoid may reduce the impact of the evolutionary trap for indigenous parasitoid species. There is a possibility that the occurrence of facultative hyperparasitism between scelionid parasitoids associated with stinkbugs is common. This resulting intraguild predation could promote conservation and stabilization of natural communities by impacting the diversity and population dynamics of native stinkbugs and their parasitoids (e.g., by allowing native parasitoids to avoid wasting reproductive effort on unsuitable hosts), or reduce success of biological control programs (e.g., by reducing the population size of the exotic parasitoids).

Overwintering insects must survive the multiple-stress environment of winter, which includes low temperatures, reduced food and water availability, and cold-active pathogens. Many insects overwinter in diapause, a developmental arrest associated with high stress tolerance. Drosophila suzukii (Diptera: Drosophilidae), spotted wing drosophila, is an invasive agricultural pest worldwide. Its ability to overwinter and therefore establish in temperate regions could have severe implications for fruit crop industries. We demonstrate here that laboratory populations of Canadian D. suzukii larvae reared under short-day, low temperature, conditions develop into dark 'winter morph' adults similar to those reported globally from field captures, and observed by us in southern Ontario, Canada. These winter-acclimated adults have delayed reproductive maturity, enhanced cold tolerance, and can remain active at low temperatures, although they do not have the increased desiccation tolerance or survival of fungal pathogen challenges that might be expected from a more heavily melanised cuticle. Winter-acclimated female D. suzukii have underdeveloped ovaries and altered transcript levels of several genes associated with reproduction and stress. While superficially indicative of reproductive diapause, the delayed reproductive maturity of winter-acclimated D. suzukii appears to be temperature-dependent, not regulated by photoperiod, and is thus unlikely to be 'true' diapause. The traits of this 'winter morph', however, likely facilitate overwintering in southern Canada, and have probably contributed to the global success of this fly as an invasive species. (C) 2016 Elsevier Ltd. All rights reserved.

1 Peristenus digoneutis Loan is a parasitoid of Lygus plant bugs, which was successfully introduced from Europe into North America in the 1980s for controlling native Lygus populations. Surveys confirmed that P. digoneutis populations have become established throughout eastern North America and that the spread of the parasitoid continues. For unknown reasons, previous releases of P. digoneutis in Western Canada were not successful. 2 A bioclimate (climex®; Hearne Scientific Software Pty Ltd, Australia) model for P. digoneutis in North America was developed, based on climate and ecological parameters, and then validated with actual distribution records. The current distribution of P. digoneutis in eastern North America was consistent with the predicted distribution. The model suggests that P. digoneutis will probably continue its spread westwards throughout the U.S.A. along the Great Lakes. 3 The southern distribution of P. digoneutis is expected to be limited by hot summer temperatures, whereas its northern range is limited by the number of Lygus host generations rather than cold stress. 4 Peristenus digoneutis has the potential to occur in the southern parts of the prairie ecozone of western Canada; however, Ecoclimatic Index values in the prairies indicate mainly marginal or unfavourable conditions, which may explain why earlier releases of P. digoneutis in Western Canada failed.

Ticks are among the most important vectors of disease in the Northern Hemisphere, and a better understanding of their feeding behaviour and life cycle is critical to the management and control of tick-borne zoonoses. DNA-based tools for the identification of residual bloodmeals in hematophagous arthropods have proven useful in the investigation of patterns of host use in nature. Using a blind test approach, we challenged the utility of the DNA barcode library for the identification of vertebrate bloodmeals in engorged, field-collected Ixodes scapularis. Universal vertebrate primers for the COI barcode region successfully amplified DNA from the host bloodmeal and only rarely amplified tick DNA. Of the 61 field-collected ticks, conclusive genus- and species-level identification was possible for 72% of the specimens. In all but two cases, barcode-based identification of the bloodmeal was consistent with the morphological identification of the vertebrate host the ticks were collected from. Possible explanations for mismatches or ambiguities are presented. This study validates the utility of the DNA barcode library as a valuable and reliable resource for the identification of unknown bloodmeals in arthropod vectors of disease. Future directions aimed at the refinement of these techniques to gain additional information and to improve the amplification success of digested vertebrate DNA in tick bloodmeals are discussed.

LygusHahn plant bugs (Hemiptera: Miridae) are serious pests of a wide variety of economically important crops in North America. EuropeanPeristenus digoneutisLoan andP. relictusRuthe (Hymenoptera: Braconidae) are being considered for release in Canada as part of a classical biological control program forLygus. The attractiveness of different host plants to EuropeanPeristenushas not been addressed, but may be an important consideration prior to parasitoid release.Lygus rugulipennisPoppius nymphs were collected in the Northern Temperate Atlantic (NTA) ecoregion on red clover (Trifolium pratenseL.; Fabaceae) and chamomile (Matricaria recutitaL.; Asteraceae), and in the Western European Broadleaf Forest (WEBF) ecoregion on red clover and alfalfa (Medicago sativaL.; Fabaceae). Parasitism levels and parasitoid species were determined using a multiplex PCR assay forP. digoneutis,P. relictus, andP. pallipesCurtis. Mean parasitism levels inL. rugulipenniswere 45–49% in the NTA ecoregion and 25–32% in the WEBF ecoregion. However, in neither ecoregion were parasitism levels and parasitoid species compositions significantly different in nymphs from different host plant species. Furthermore, multiparasitism was low despite the fact thatP. digoneutisandP. relictusshare the same host species.

Summary 1 Accurate identification of natural enemies is the cornerstone of biological control, and methods that can separate closely related species are essential in ecological studies of parasitoids. Conventionally, host rearing and dissection are used to define the ecological host range of candidate biological control agents and assess host-specificity of parasitoids. However, molecular methods may be more suitable for the evaluation of host–parasitoid associations. 2 To demonstrate the utility of molecular diagnostics in ecological host-range studies on parasitoids of Lygus plant bugs, host rearing, dissection and multiplex polymerase chain reaction (PCR) analysis were used to estimate parasitism levels and parasitoid species composition (genus Peristenus) in more than 26 000 field-collected target and non-target Miridae. 3 Parasitism levels estimated by conventional and molecular methods were similar but molecular analysis can detect parasitoids earlier than dissection and rearing. Parasitoid pupal mortality prevented the identification of more than 30% of individuals reared from non-target host material; however, paired samples analysed with the multiplex assay allowed the identity of these parasitoids to be inferred. Molecular methods can provide different, and generally more complete, parasitoid species composition information because the results are not confounded by the host and parasitoid mortality encountered in rearing. However, detection of a parasitoid in a host does not necessarily indicate survival to the adult stage. Further, molecular identification of parasitoid species may be restricted to those species for which PCR primers are available. 4 Synthesis and applications. For molecular diagnostic techniques to gain widespread adoption in ecological studies on natural enemy host range, they must provide information that is equivalent (or superior) to information obtained by conventional methods. Based on a large-scale case study, associations between Peristenus spp. and their mirid hosts were used to demonstrate the utility of molecular diagnostics in studies on parasitoid ecological host range; however, this approach can be extended to pre-release risk-assessment studies on other candidate biological control agents. Beyond agent identification, molecular diagnostics can facilitate and expedite pre- and post-release studies on the ecological host range of parasitoids, potential non-target effects, host–parasitoid associations and trophic interactions.

Lygus spp. Hahn (Hemiptera: Miridae) are serious pests of a wide variety of economically important crops in North America. A European parasitoid, Peristenus digoneutis Loan (Hymenoptera: Braconidae), released in the northeastern USA for the biological control of Lygus, has successfully established in nine states and in eastern Canada, including southeastern Ontario, southern Quebec and Nova Scotia. To determine the extent to which P. digoneutis has dispersed and established in mirid populations in Ontario, a single-step multiplex PCR assay, designed to differentiate European species P. digoneutis and P. relictus from native North American Peristenus species, was used to identify parasitoid larvae dissected from field-collected Miridae. A total of 222 parasitoid larvae were analysed with the multiplex assay. Most (172) were identified as members of the P. pallipes Curtis complex; however, three from the Niagara region were identified as P. digoneutis. Specimens that did not amplify using the Peris...

The polymerase chain reaction (PCR) revolutionized the field of diagnostics, and today it has routine applications in medical, veterinary, forensic and botanical sciences. The fields of biological control and insect pest management have generally been slow to adopt PCR-based diagnostics in comparison with other fields of science. However, there has been increasing interest in the use of molecular diagnostic tools in arthropod biological control. In applied entomology, molecular techniques have generally been used for insect identification and systematics; however, PCR-based techniques are increasingly becoming recognized as valuable tools in ecological studies. Here, we review research that has used PCR-based techniques for parasitoid and predator/prey identification and detection, and place these studies in the context of their contributions to biological control of arthropods. The status and future directions of diagnostic molecular markers in applied entomology and insect pest management are also discussed.

International audience; The brown marmorated stink bug, Halyomorpha halys (Hemiptera: Pentatomidae), has emerged as a harmful invasive insect pest in North America and Europe in the 1990s and 2000s, respectively. Native to eastern Asia, this highly polyphagous pest (> 120 different host plants) is spreading rapidly worldwide, notably through human activities. The increasing global importance of the pest suggests that more coordinated actions are needed to slow its spread and mitigate negative effects in invaded areas. Prevention of large-scale outbreaks will require accurate identification and effective mitigation tools to be rapidly developed and widely implemented. In this short review, we update the current distribution of H. halys, discuss potential geographic range expansion based on passive and active dispersal and provide insight on the economic, environmental and social impact associated with H. halys.

Lygus Hahn (Hemiptera: Miridae) are serious pests of agricultural and greenhouse crops throughout North America. In Europe, bivoltine Peristenus Forster (Hymenoptera: Braconidae) species have a significant impact on Lygus populations. Release and establishment of European P. digoneutis Loan in Lygus lineolaris (Palisot de Beauvois) populations in northeastern USA has renewed interest in the intended liberation of European parasitoids for Lygus control in Canada. Accurate identification of natural enemies is the cornerstone of biological control but conventional methods for identifying Peristenus species and estimating parasitism rates rely on tedious and time-consuming dissection and rearing methods. The present study describes species-specific PCR primers for three species of Peristenus, and the use of a multiplex PCR assay to detect P. digoneutis and P. stygicus Loan eggs and larvae from Lygus rugulipennis Poppius nymphs. Results indicate that the primers amplify uniquely sized, species-specifi...

Species-specific primers for four species of Neofabraea associated with apple and pear fruit diseases were used in multiplex PCR assays to identify 29 putative Neofabraea isolates, primarily isolated from bull's-eye rot on pears from packing houses in Washington and Oregon. Apples were inoculated with these isolates, and tentative identifications based on morphology of conidia forming on wound-inoculated fruits were compared with identifications made using species-specific primers for different Neofabraea species. PCR-based identifications were successful for 26 of 29 isolates, and in all cases were consistent with tentative identifications based on spore morphologies. The results revealed that in addition to the two species of Neofabraea already known to occur in the Pacific Northwest, N. malicorticis and N. perennans, a third species, N. alba, is also prevalent.