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Journal of Pest Science (2019) 92:281–297 https://doi.org/10.1007/s10340-018-0997-6

ORIGINAL PAPER

Multi‑component blends for trapping native and exotic longhorn at potential points‑of‑entry and in forests

Jian‑ting Fan1,2 · Olivier Denux1 · Claudine Courtin1 · Alexis Bernard1 · Marion Javal1 · Jocelyn G. Millar3,4 · Lawrence M. Hanks5 · Alain Roques1

Received: 14 February 2018 / Revised: 26 May 2018 / Accepted: 28 May 2018 / Published online: 7 June 2018 © Springer-Verlag GmbH , part of Springer Nature 2018

Abstract The accidental introduction of exotic wood-boring cerambycid beetles represents an ever-increasing threat to forest biosecu- rity and the economies of many countries. Early detection of such upon arrival at potential points-of-entry is challeng- ing. Because pheromone components are often conserved among related species in the family Cerambycidae, we tested the generic attractiveness of diferent blends of pheromones composed of increasing numbers of pheromone components at both potential points-of-entry and in natural forests in during 2014–2017. Initially, two diferent four-component blends were compared, one composed of fuscumol, fuscumol acetate, geranylacetone, and monochamol, and the other composed of 3-hydroxyhexan-2-one, anti-2,3-hexanediol, 2-methylbutanol, and prionic acid. In a second step, host volatiles (ethanol and [-]-α-pinene) were added, and fnally, we tested the efectiveness of a mixture of all eight pheromone components with the two host volatiles. Overall, 13,153 cerambycid beetles of 118 species were trapped. The 114 native species trapped represent 48% of the French fauna, including more than 50% of the species in 25 of the 41 cerambycid tribes. At potential points-of-entry, captures included 2960 cerambycids of 49 species, including three exotic Asian species, two of which had never been reported previously in . In forests, attraction to the four-component blends varied with their composition. Adding host volatiles did not change the overall attraction except for the species testaceus, which showed a fourfold increase in captures. Placing the two four-component blends on the same trap resulted in signifcant increases in the number of species and individuals captured compared to captures by traps baited with each blend individually. Finally, the eight-component pheromone blend was found to be as attractive as the combination of the two four-component blends hung together on the same trap, without apparent antagonistic efects. This fnding suggests that use of multi-component lures may help to minimize the costs and manpower required to detect exotic and potentially invasive species.

Keywords Cerambycidae · Early detection · Multi-component pheromone lures · Exotic · Trapping · Ports

Key message

Communicated by J.D. Sweeney. • Early detection of exotic cerambycid beetles upon arrival at points-of-entry is a major challenge for regulatory Special Issue on Invasive Pests of Forests and Urban Trees. agencies. Electronic supplementary material The online version of this • The parsimony in pheromone structures used by a num- article (https​://doi.org/10.1007/s1034​0-018-0997-6) contains ber of cerambycid species worldwide suggested that supplementary material, which is available to authorized users.

* Alain Roques 3 Department of Entomology, University of California, [email protected] Riverside, CA 92506, USA 4 Department of Chemistry, University of California, 1 INRA UR 633 Zoologie Forestière, 2163 Avenue de la Riverside, CA 92506, USA Pomme de Pin, 45075 Orléans, France 5 Department of Entomology, University of Illinois 2 School of Forestry and Biotechnology, Zhejiang Agriculture at Urbana-Champaign, Urbana, IL 61801, USA and Forestry University, Lin’an, China

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multi-component blends of pheromones could be used lure combinations improve trapping efciency without sig- as efective trap baits, while minimizing costs and man- nifcant antagonistic efects. power. To date, most trapping trials carried out within or near to • Results showed that the number of species detected at a ports and other points-of-entry for international cargo pri- site was maximized by a blend of eight pheromone com- marily have tested single lures rather than blends of attract- ponents, together with plant volatiles, without signifcant ants (Brockerhof et al. 2006) and have focused on bark bee- antagonistic interactions among components. tles rather than cerambycids (Rabaglia et al. 2008). However, • Trapping of native species around ports and airports indi- Rassati et al. (2014) compared attraction of bark beetles to cated their potential to be moved as exports. single versus multiple components in Italian seaports and found that the multi-lure trap was as efective as the sum of the single-lure traps, with no evidence of any negative interactions among the tested compounds, suggesting that Introduction efective monitoring with multi-lure traps may be feasible. Recent advances in the identifcation of cerambycid pher- Exotic wood-boring beetles, especially those in the families omones are now ofering analogous opportunities for devel- Cerambycidae and Buprestidae, and the subfamily Scolyti- oping multi-lure blends aimed at attracting multiple species nae of the Curculionidae, represent serious threats to for- simultaneously (Wong et al. 2012). Sex and aggregation-sex est biosecurity and the economies of all forested countries. pheromones are known for several hundred cerambycid spe- These usually arrive as immatures sequestered within cies (reviewed by Millar and Hanks 2017), and pheromone wood, wood products, or wooden packing material such as structures are frequently highly conserved among related pallets, crating, and dunnage (Brockerhof et al. 2006; Lieb- species (Hanks and Millar 2013, 2016). For example, pher- hold et al. 2012; Rassati et al. 2014), and increasingly, ship- omones of many species in the subfamily ments of nursery plants and bonsais (Roques 2010; Liebhold are composed of 3-hydroxyalkan-2-ones, 2-hydroxyalkan- et al. 2012). Therefore, air, sea, and river ports, and associ- 3-ones, or 2,3-alkanediols, whereas pheromones of many ated facilities for handling and storing international cargos, species in the subfamily consist of hydroxyethers are likely points-of-entry for exotic wood-boring beetles. and related compounds, the terpenoid fuscumol ([E]- The problem is exacerbated by the fact that newly observed 6,10-dimethylundeca-5,9-dien-2-ol) and its corresponding introductions and establishments usually are due to “emerg- acetate, while fuscumol also is used as a pheromone by spe- ing” species, i.e., those that have never been reported as cies in the subfamily (reviewed in Hanks and introduced elsewhere, and which are not signifcant pests in Millar 2016). Furthermore, the use of the same compounds their native range (Seebens et al. 2018). Such species usually by species on multiple continents (e.g., Wickham et al. 2014 are not subject to regulatory measures because their invasive in China; Sweeney et al. 2014 in ; Hayes et al. 2016 potential has not been recognized. The development of new in Australia; Silva et al. 2017 in Brazil) has demonstrated strategies to detect such unanticipated, unregulated species the potential for exploiting such compounds to detect exotic as early as possible is essential in order to implement rapid species upon their arrival on another continent. and efective eradication measures (Rassati et al. 2014). Species which share dominant pheromone components Countries such as Australia (Bashford 2008), New Zea- appear to use several mechanisms to avoid cross-attraction, land (Brockerhof et al. 2006), the USA (Rabaglia et al. such as difering in seasonal and/or daily activity period, 2008), and (Rassati et al. 2015) have recently imple- or use of minor pheromone components to create species- mented strategies to enhance the early detection of alien specifc blends (Hanks and Millar 2013; Mitchell et al. 2015; wood-boring beetles based on baited traps deployed in ports Meier et al. 2016). These fndings have resulted in the devel- or other high-risk sites. Such traps should ideally be ef- opment and optimization of methods for using pheromone- cient at low population densities and should target multiple based attractants to detect many cerambycid species and to species of diverse taxa simultaneously, both because it is characterize cerambycid communities in North America impossible to predict which species may arrive (Rassati et al. (Graham et al. 2010; Allison et al. 2011, 2014; Mitchell 2014) and because costs escalate rapidly with increasing et al. 2011; Hanks and Millar 2013; Handley et al. 2015; numbers of traps deployed. Moreover, because the practi- Webster et al. 2016). calities of installing traps in safe and suitable places in such Several experiments carried out in natural landscapes in sites of high activity are limiting, it is essential to minimize North America have shown that use of multi-component the number of traps within each port-of-entry. Traps baited blends of cerambycid pheromones as trap baits may be with blends of pheromones and related attractants would both feasible and efective (Hanks et al. 2012, 2018; Millar thus represent a major improvement in currents protocols et al. 2018). Partial inhibition was noted for a few species, (Brockerhof et al. 2006; Hanks et al. 2012) but only if the with captures being higher in traps baited with a specifc

1 3 Journal of Pest Science (2019) 92:281–297 283 compound than in those baited with a blend containing that in forests were hung in the lower canopy (bottom of the compound. Nevertheless, the authors suggested that such trap 1.5–3 m above the ground). The supplied trap basins interactions could be minimized by judicious choice of com- were modifed, with the bottom replaced by a wire mesh to ponents to include in a blend, and furthermore, that as long allow drainage to keep specimens dry. An insecticide sachet as inhibition did not completely prevent attraction, one trap (Ferag­ ®, SEDEQ, ) was added to kill the trapped speci- with a multi-component lure would still be much more cost- mens. It was replaced in 2016–2017 with netting impreg- efective than deploying multiple traps baited with single nated with alpha-cypermethrin insecticide (Storanet­ ®, BASF lures. Pfanzenschutz Deutschland, Germany). Trap panels and the The technical and legal requirements associated with interior parts of collection basins were treated each year with ports, airports, and other potential points-of-entry present the lubricant fuon (AGC Chemicals Europe Ltd., Thornton considerable constraints compared to monitoring ceram- Cleveleys, UK) diluted in 1:6 in water, to improve trapping bycids in natural landscapes. They include limitations on efciency (Graham et al. 2010). In all years, trials were con- trap placement (e.g., to avoid destruction by cargo handling ducted from late April to mid-October. equipment), trap maintenance (e.g., traps easily serviced by Pheromones and attractants to be included in multi- non-professionals), -conservation systems, and the species lures were selected so as to cover a large number need to kill and preserve specimens for molecular analyses of cerambycid subfamilies and tribes according to data in to confrm identity as possible exotic species (e.g., killed dry Hanks et al. (2012). We used a progressive combination of using insecticides). At points-of-entry, it is also impossible the blends during the four years of the study, with a one-year to predict which species might be entering, and low trap delay between similar tests in forests and potential points- catches may be difcult to analyze statistically. Thus, paral- of-entry (Table 1). The frst tested lures consisted of two dif- lel tests in forest stands are necessary to assess the efective- ferent blends and a control. Blend #1 included 50 mg of fus- ness of pheromone blends before they can be recommended cumol, 50 mg of fuscumol acetate, 50 mg of monochamol, for use in surveillance of exotic species. and 25 mg of geranylacetone dissolved in isopropanol as a Therefore, our goals were: (1) to evaluate the generic carrier to a total volume of 1 ml per lure. Blend #2 contained attractiveness of several multi-component blends of ceram- 50 mg of 3-hydroxy-2-hexanone, 1 mg of prionic acid (race- bycid pheromones for the native cerambycid fauna in stands mic 3,5-dimethyldodecanoic acid), 50 mg of 2-methylbutan- representative of the diversity of French forests; (2) to test, 1-ol, and 50 mg of anti-2,3-hexanediol (hence referred to through successive annual experiments, the comparative ef- as 2,3-C6-diol) dissolved in isopropanol as a carrier to a cacy of a combination of these blends up to an eight-com- total volume of 1 ml per lure. Control lures (#T) contained ponent blend; and (3) to carry out tests of multi-component 1 ml of isopropanol. All compounds were purchased from blends within diferent types of potential points-of-entry ChemTica Internacional, S.A. (Heredia, Costa Rica) except (airports, ports, markets), with traps placed both on site and prionic acid (Alpha Scents Inc.), and anti-2,3-hexanediol in suitable habitats within a radius of 1 km from the likely used in 2014, which was synthesized as described in Lacey point of arrival so as to monitor dispersal of beetles away et al. (2004). Emitters were clear polyethylene sachets (Mini- from the site of introduction, while simultaneously assessing grip, 4 cm × 6 cm × 60µ; Dutscher, Brumath, France) that whether surrounding forests might serve as sources of native were hung in the center of traps, at the top of the window species that could infest exported goods. opening. The release rate of blends determined by mass loss under 20 °C conditions was 0.018 ± 0.002 g/d for blend #1 and 0.019 ± 0.002 g/d for blend #2. Materials and methods Next, we tested the efect of adding host plant volatiles (HV) to these blends. Lure #1 + HV consisted of blend #1 Field trapping experiments were conducted during to which was added host volatiles, including 5 ml of 95% 2014–2017 in forests and during 2015–2017 in potential ethanol (release rate 0.066 ± 0.002 g/d, Carlo Erba Reagents, points-of-entry. Val de Reuil, France) and 2 ml of (-)-α-pinene (release rate 0.024 ± 0.008 g/d, Sigma-Aldrich Chimie Sarl, Saint Quen- Trapping methods tin Fallavier, France), both in separate polyethylene sachets (Minigrip 6 × 8 cm and 4 × 6 cm, respectively) hung in the Traps consisted of black intercept cross-vane panel traps center of traps but at the base of the window opening. Lure (Alpha Scents Inc., West Linn, Oregon, USA) that were sus- #2 + N consisted of blend #2 to which was added 1 ml of pended from branches of trees in forests and from various 2-nonanone, suggested as a possible attractant for inva- metallic structures in the potential points-of-entry. Because sive Anoplophora spp. (JT Fan, unpub data), in a separate it was usually impossible to hang traps at heights higher 4 × 6 cm polyethylene sachet as above. Additional lures than 3 m in the latter sites, for consistency, traps deployed tested included a combination of lure #1 + HV and lure #2

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Table 1 Trapping design used Year 2014 2015 2016 2017 in forests (F) and potential points-of-entry (P) from 2014 Site F F P F P F P to 2017 Lure #1 x x x #1 + HV x x x x #2 x x x x x #2 + N x x #1 + HV and #2 x x x x #3 + HV x #T x x x

Lures: #1: fuscumol, fuscumol acetate, monochamol, and geranylacetone in isopropanol; #2: 3-hydroxy- 2-hexanone, prionic acid, 2-methylbutan-1-ol, and anti-2,3-hexanediol in isopropanol; #3: blend #1 and blend #2 combined in isopropanol; #T: isopropanol; HV: (-)α-pinene and ethanol in separate bags; N: 2-nonanone in separate bags

(1 ml) hung side-by-side in separate sachets on the same but were rotated in 2017 to control for location efects by trap, and fnally a multi-lure blend #3 which contained the moving entire traps at the time they were serviced. The tim- eight pheromonal compounds described above (1 ml; release ing of lure and insecticide replacement as well as collection rate estimated to 0.0263 ± 0.002 g/d), which was paired with of trap contents was as before. 5 ml of ethanol and 2 ml of (-)-α-pinene in separate poly- In 2014, we compared the attractiveness of two diferent ethylene sachets. multi-lure blends (#1 and #2) with regard to that of a con- Lures and insecticides were replaced every 3 wk, at which trol (#T; Table 1), using all the previous sites for a total of time trapped beetles were collected into 95% ethanol for 22 replicates (Table 2). The total number of traps was thus identifcation. Captured beetles were identifed according 66. The same blends were again tested in 2015, but host to Bense (1995) and Berger (2012). The sexes of trapped plant volatiles (HV) were added in order to trap bark beetles cerambycids were not recorded, because the pheromones simultaneously with cerambycids. This resulted in fve dif- and attractants used generally attracted adults of both sexes ferent treatments: #1, #1 + HV, #2, #2 + N, #T. Due to fund- in similar numbers (Millar and Hanks 2017), although male- ing constraints, the 2015 trials could only be deployed at 5 produced aggregation pheromones could result in female- of the 2014 sites, for a total of nine replicates in the southern biased captures in some species (Silva et al. 2017), and the French Alps, north-central France, and south-central France primary objective of this study was to determine the ef- (Table 2). A total of 45 traps was deployed. ciency of the blends for species detection, regardless of sex. Given the 2015 results, we retained only lure #1 + HV All the trapped specimens are preserved at the INRA Forest and lure #2 for tests in 2016. These tests aimed at comparing Zoology Research Unit, Orléans, France. the specifc attractiveness of traps combining lure #1 + HV and lure #2, but hung side-by-side in separate sachets on the Trapping in forests same trap, versus attractiveness of traps baited with each of these two multi-lures separately. Trials were conducted in The experiments aimed at covering most of the woody vege- 10 of the 2014 sites, for a total of 20 replicates, with a few tation types existing in France and the largest possible diver- changes in the number of replicates per area (Table 2). In sity of cerambycid species native to France. Field trials thus each replicate, three traps were deployed with either lure were deployed from north-central France to the Mediter- #1 + HV, lure #2, or the combination of these two lures. ranean coast and Corsica, including mountainous areas (the The total number of traps was thus 60. At two sites (north- Alps, Massif Central; Table 2). This design encompassed central France and south-central France), prionic acid was most of the native species (Pinus halepensis, P. nigra removed from lure #2 because in 2015, lures containing laricio, P. pinaster, P. sylvestris, P. uncinata), junipers (Juni- prionic acid attracted hundreds of large coriarius perus communis, J. thurifera), and species (Quercus which destroyed most other specimens, resulting in identi- petreae, Q. robur, Q. suber) for a total of 14 sites with 1–4 fcation difculties. replicates per site depending on the year (Table 2). In each In 2017, we compared the attractiveness of traps baited replicate, 1 trap per lure was deployed, with traps positioned with the combination of lure #1 + HV and lure #2, hung at least 12 m apart in linear transects. Treatments were not side-by-side in separate sachets on the same trap with rotated among positions during the trials from 2014 to 2016, that of the 8-component blend, lure #3 described above

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Table 2 Study sites used for feld experiments from 2014 to 2017 in forests and at potential points-of-entry in France, with details of their char- acteristics Region/site Latitude, longitude Habitat type No. of replicates 2014 2015 2016 2017 Forests Southern French Alps-Montgenèvre 44.925639°, 6.698083° Mature forest of mountain pine (Pinus 2 1 1 1 uncinata) Southern French Alps-Névache 44.988793°, 6.668460° Mature forest of Scots pine (Pinus sylves- 2 1 1 1 tris) Southern French Alps-Saint Crépin 44.710290°, 6.606683° Mature forest of incense juniper (Junipe- 1 – – – rus thurifera) Southern French Alps-Villard Saint 44.861056°, 6.601889° Mature forest of European (Larix 1 1 1 1 Pancrace decidua) North-Central France-Orléans 47.828457°, 1.913659° Mixed -deciduous forest of Scots 3 3 3 3 pine, sessile oak (), and pedunculated oak (Q. robur) Southern France-Gignac 43,635416°, 3,582489° Mixed conifer-deciduous forest of Aleppo 1 – 4 4 pine (Pinus halepensis) and downy oak () Southeastern France-Noves 43.751177°, 5.003673° Mixed conifer-deciduous forest of Aleppo 1 – 2 2 pine (Pinus halepensis) and downy oak (Quercus pubescens) Central France-Lavercantière seed orchards 44.579970°, 1.366446° Maritime pine (Pinus pinaster) seed 1 – – – orchard Central France-Lavercantière seed orchards 44.619007°, 1.349133° Black pine () seed orchard 1 – – – Central France-Latronquière seed orchards 44.851067°, 2.106800° () seed orchard 2 – 3 3 South-central France-Salles La Source 44.437000°, 2.555550° Mixed forest of downy oak, black pine, 3 3 3 3 and common juniper (Juniperus com- munis) Northern Corsica-Furiani 42.680733°, 9.431033° Urban forest of cork oak (Quercus suber) 1 – 1 1 Southern Corsica-Sainte Lucie de Porto 41.697317°, 9.385400° Mediterranean maquis with cork oak, 2 – 1 1 Vecchio myrtle (Myrtus communis), and pistachio ( lentiscus) Southern Corsica-Ospédale 41.653167°, 9.180150° Mature forest of Corsican pine (Pinus 1 – – 1 nigra laricio) Points-of-entry Bayonne 43.530102°, − 1.510933° Maritime port – 2E 2E/2I 2I/2E Bordeaux 44.913137°, − 0.538797° Maritime port – – – 2I/2E Fos 43.409675°, 4.841009° Maritime port – 2I 2I/2E 2I/2E La Rochelle 46.160391°, − 1.222321° Maritime port – 2I/2E 2I/2E 2I/2E Marseille 43.354296°, 5.322368° Maritime port – 2I/2E 2I/2E 2I/2E Nice 43.693963°, 7.281823° Maritime port – 2I/2E 2I/2E – Port-Vendres 42.515754°, 3.106575° Maritime port – 2I 2I/2E 2I/2E Saint Malo 48.637323°, − 2.028058° Maritime port – 2I/2E 2I/2E 2I/2E Huningue 47.595454°, 7.589142° River port – 2I/2E 2I/2E 2I/2E Châteauroux 46.860488°, 1.721526 Airport – 2I/2E 2I/2E 2I/2E Orly 48.715856°, 2.340999° Airport – 2I – – Roissy-Charles de Gaulle 49.004382°, 2.533034° Airport – – 2I/2E 2I/2E Toulouse 43.614144°, 1.383150° Airport – – – 2I/2E Rungis 48.764781°, 2.345756° National trade market – 2I 2I/2E 2I/2E Vernouillet 48.966728°, 1.976490° Arboriculture production – – 2I/2E – Roissy 49.010444°, 2.485672° Tree nursery – – – 2I/2E La Turbie 43.744300°, 7.377570° Terrestrial border – – – 2I/2E

In points-of-entry, trap replicates were deployed within the site (I) and/or within a 1 km-radius (E)

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(including prionic acid), which was paired with ethanol and replaced the arboricultural site (Table 2). Two traps were (-)-α-pinene in separate polyethylene sachets. The lures were deployed within the site and two others nearby for a total tested both separately and as one mixture because interac- of 60 traps. tions among the components may exist in the 8-component blend. The same areas as in 2016 were used for a total of Data analysis 21 replicates, with the addition of a replicate at Ospédale, Corsica. Each replicate consisted of two traps, one with the For each annual experiment in forests, overall lure efects combination of lure #1 + HV and lure #2 in separate sachets were tested by comparing both the total number of ceramby- and one with lure #3 + HV. cid species and the total number of individuals captured per blend at each site. Because traps were not rotated in 2014, Trapping at potential points‑of‑entry 2015, and 2016, the number of replicates was based on the number of sites. Trap rotation in 2017 in forests allowed us These trials used 17 sites covering a variety of potential to consider the number of collection dates at each site as points-of-entry for cerambycid invaders, including seven replicates. Replicates from a given date that contained no maritime ports, one river port, two airports, and one national cerambycids in any of the traps, for example due to inclem- market (Table 2). The selected sites corresponded to key ent weather, were dropped from the analyses. Because data trade and tourism hubs in France, some being important for violated normality, diferences between trap captures were the trade of timber and wood products (e.g., La Rochelle tested using the nonparametric Friedman’s Q test (Statistica for timber imported from ), others depending on the ­9®, Tibco Software Inc., Palo Alto, CA, USA). Assuming a interest of port authorities. At each site, traps were deployed signifcant overall Friedman’s test, pairs of treatment means within the port, as close as possible to places where wood were compared with the nonparametric Dunn–Nemenyi and/or its byproducts were stored, or near wood-waste land- multiple comparison test. The efect of lures on individual flls, and traps were placed in nearby wooded areas within species was similarly tested, but we analyzed data only for a radius of 1 km in order to survey the presence of species species that were represented by at least 10 specimens dur- similar to the ones trapped within the port and assess pos- ing the entire trapping season. Although the captures were sible movements from and to the port. much lower than in forests, the same analysis was applied The 2014 results in forests led us in 2015 to test in both to trapping results in points-of-entry in 2015 and 2016, but forests and potential points-of-entry the same fve difer- not in 2017 because only one lure was used during this year. ent treatments: #1, #1 + HV, #2, #2 + N, #T. A total of 11 Using Student t test for paired samples, species captures potential points-of-entry were considered (Table 2). At each by subfamilies and tribes of Cerambycidae in forests were site, we had intended to deploy 10 traps (two per lure), fve compared in 2016 between traps baited with the lure combi- within the port and fve in a nearby wooded area. However, nation (lure #1 + HV and lure #2 hung side-by-side in sepa- it proved impossible to deploy traps outside four potential rate sachets) and the sum of the captures in separate traps points-of-entry (Orly, Rungis, Port-Vendres, Fos), and we baited with either lure #1 + HV or lure #2. The same test did not get authorization in time to deploy traps within the was applied in 2017 to compare captures by the multi-lure port of Bayonne and thus only deployed traps outside the blend #3 and the lure combination #1 + HV and lure #2 hung port. A total of 65 traps were thus fnally deployed. side-by-side in separate sachets. Given the 2015 results, we retained only lure #1 + HV and lure #2 for tests in 2016. Trials were conducted at 12 potential points-of-entry, the same ones as in 2015 except Results that Roissy-Charles de Gaulle airport replaced Orly airport, and a site dedicated to arboricultural production near Paris Throughout the four-year course of this study, a total of was added (Table 2). Four traps were deployed within each 13,153 cerambycids representing 118 species were caught. site with two replicates per lure, and four traps in the vicin- ity of the site, also with two replicates per lure. The total Trapping in forests number of traps was thus 96. In 2017, traps were only baited with the combination Overall, 107 species were trapped in forests, totaling 10,193 of lure #1 + HV and lure #2 hung side-by-side in separate specimens. These included 105 native and two exotic spe- sachets on the same trap. These traps were deployed at 14 cies, stebbingi and the Australian ports of entry, including the addition of Toulouse airport semipunctata (one specimen; Table 3). The 2014 trials cap- and the terrestrial border road between Italy and France at tured a total of 2434 cerambycids in 59 species. These tests La Turbie, whereas the maritime port of Bordeaux replaced revealed signifcantly diferent patterns of attraction between that of Nice and a forest tree nursery near the Roissy airport lures in both the mean number of species trapped per site

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Table 3 Subfamily, tribe, species, and total numbers of cerambycid #T: isopropanol; #1 + HV: blend #1, (-)α-pinene and ethanol in three beetles attracted per lure during 2014–2017 within and in the vicinity separate bags; #2 + N: blend #2 and 2-nonanone in two separate bags; of potential points-of-entry and in French forests. Exotic species are #(1 + HV) + #2: blend #1, (-)α-pinene, ethanol and blend #2 in four underlined. Lures: #1: fuscumol, fuscumol acetate, monochamol, and separate bags; #3: + HV: blend #1 and blend #2 combined in isopro- geranylacetone in isopropanol; #2: 3-hydroxy-2-hexanone, prionic panol, and (-)α-pinene and ethanol in three separate bags acid, 2-methylbutan-1-ol, and anti-2,3-hexanediol in isopropanol; Grand Ports Forests Total Within Near Total Total 2014 2015 2016 2017 port Port ports Forests Lure #1 #2 #T #1 #1+HV #2 #2+N #T #1+HV #2 #(1+HV)+#2 #(1+HV)+#2 #3+HV Species Cerambycinae Anoplodera rufipes - - - 2 ------2 2 abdominalis ------3 3 3 Callidium aeneum - - - 2 ------3 12 1 18 18 Callidium coriaceum ------3 5 - - 8 8 bajulus 14 13 27 ------2 - - 2 4 31 Leioderes kollari ------1 - 1 1 461 930 1391 4 410 1 - - 212 122 - 7 330 1351 155 198 2790 4181 Poeciliumfasciatum 1 - 1 ------1 Poeciliumalni 11 118 129 2 76 4 - - 383 71 - 1 517 245 18 8 1325 1454 Pyrrhidium sanguineum 7 45 52 - 210 6 - - 71 13 - 5 459 434 59 71 1328 1380 cerdo ------1 - 2 3 - 6 6 Cerambyx miles ------3 - - - - 3 3 Cerambyx scopolii ------1 - 1 1 Cerambyx welensii ------1 - - 1 1 Certallini Certallum ebulinum ------1 - - - 1 1 0 Anaglyptus gibbosus ------2 3 - - 5 5 s - - - - 4 ------3 8 1 - 16 16 i 1 - 1 ------1 figuratus ------1 1 2 2 Chlorophorus glabromaculatus 10 51 61 - 2 2 - - 1 - - 2 4 5 14 16 46 107 Chlorophorus glaucus - - - - 1 ------1 1 Chlorophorus pilosus - 1 1 ------1 - - - - 1 ------1 1 Chlorophorus trifasciatus - - - - 1 ------1 1 Chlorophorus varius ------1 - - - 1 1 1 5 6 - 1 - - - - - 2 - 1 2 - 6 12 lama - - - 10 11 7 2 2 1 1 4 5 - 9 3 - 55 55 ------7 42 49 49 Clytus tropicus - - - 1 ------1 1 arcuatus 2 - 2 ------3 2 1 6 8 Plagionotus detritus - 1 1 ------1 - 4 3 8 9 s - 3 3 - 1 - - 2 ------1 1 5 8 Xylotrechus altaicus 1 - 1 ------1 e - 4 4 - 2 ------1 - - - - 3 7 Xylotrechus arvicola 9 26 35 1 5 1 - - 3 1 - - 3 2 21 18 55 90 Xylotrechus stebbingi 54 57 111 4 2 ------7 5 14 13 45 156 Deilini Deilus fugax - - - 6 3 4 ------13 13 minuta 1 3 4 ------4 fasciata ------1 ------1 1 Hesperophanes sericeus ------1 - - 1 1 Trichoferus fasciculatus - 1 1 - 1 ------1 - 2 3 Trichoferus holosericeus - - - - 1 ------1 1 Trichoferus pallidus - 1 1 ------1 Molorchus minor - - - 2 ------2 2 Nathriini brevipennis 5 6 11 - - - 1 ------1 12 Obrium brunneum ------1 ------1 1 Obrium cantharinum - 4 4 ------4 0 - - - - - 1 ------1 1 Purpuricenini budensis ------2 - 1 4 2 9 9 Purpuricenus globulicollis ------1 - 1 1 Purpuricenus koheleri ------1 - - - 1 2 2 Callimus angulatus - 2 2 ------1 - 1 3 ater 1 5 6 - 1 ------1 7

(Friedman’s Q2,21 = 12.5, P = 0.0019; Fig. 1a) and the caught 36 and 32 species, respectively, and did not difer in mean number of individuals of each species (Q2,21 = 14.4, the mean number of species trapped per site (Dunn–Neme- P = 0.0008; Fig. 1b). Traps baited with lures #1 and #2 nyi post hoc test; P = 0.77), but they only shared 15 species

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Table 3 (continued)

Stenopterus rufus 2 1 3 - - 1 - - - - - 4 4 1 - - 10 13 Lamiinae Acanthocinus aedili s ------1 1 1 Acanthocinus griseu s - 4 4 - 1 ------1 - 2 2 2 8 12 femoratu s 20 22 42 ------3 - 13 2 1 19 61 Leiopus linne i 4 15 19 ------35 - 35 2 - 72 91 Leiopus nebulosu s 70 54 124 46 - 1 - 1 - - - 1 - 2 10 10 71 195 - - clavipes 23 36 59 116 - 2 5 8 - - - 12 1 12 23 8 187 246 Aegomorphus francoe i 2 - 2 ------5 - 5 3 - 13 15 Desmiphorini Deroplia trober ------1 ------1 1 Lamia textor ------1 1 1 Uraecha angusta 1 - 1 - 1 nebulosa - - - 2 - - 2 - - - - 3 3 2 1 - 13 13 Monochamini galloprovinciali s 59 494 553 239 4 9 36 135 - - - 283 5 247 159 178 1295 1848 Monochamus sartor ------5 4 - - - 2 - 1 2 1 15 15 Monochamus suto r - - - 68 2 - 5 25 - 1 - 6 - 7 5 8 127 127 asper ------1 1 1 caroli ------1 1 1 Pogonocherus decoratu s 1 12 13 ------6 - 1 2 4 13 26 Pogonocherus hispidulu s 1 1 2 ------2 - 2 - 4 6 Pogonocherus hispidu s - 1 1 4 - 2 ------1 2 2 11 12 - 1 1 - 1 1 1 ------1 - 4 5 pecnicornis - 8 8 - - - - - 6 1 - 1 - 1 1 1 11 19 carcharias - 4 4 - 1 ------1 5 ------1 - - 1 - 2 2 ------1 ------1 1 dubi a - - - 7 1 1 1 - 1 ------1 12 12 Anastrangalia sanguinolenta ------1 2 - 3 6 6 Cortodera femorata - - - 1 - - - - - 1 ------2 2 Cortodera humeralis var. suturalis - - - 1 3 2 - 1 - 1 - - 1 - - 1 10 10 Grammoptera ruficornis - 5 5 2 ------2 7 Grammoptera ustulat a - - - - - 1 - - 1 ------2 2 Leptura aurulenta - 1 1 ------1 Pachytodes cerambyciformis ------1 1 1 Paracorymbia fulva ------2 ------2 2 Paracorymbia hybrida - - - - 1 ------1 1 Ruptela maculat a 1 5 6 3 1 - - - - - 1 3 2 1 - 3 14 20 - - - - 2 1 ------3 3 - - - 2 - 3 ------5 5 Stenurella nigr a - - - 1 ------1 1 Sctoleptura rubra - - - - - 1 - - - - - 2 - 1 - - 4 4 Sctoleptura scutellata - - - - 1 - - - - 1 - - - - - 1 3 3 Vadonia unipunctata - 1 1 ------1 Rhagiini 0 Acmaeops marginatus - - - 10 14 6 - 6 - - - 2 - 2 3 2 45 45 Acmaeops pratensi s ------14 - - 1 38 - 25 4 2 84 84 Acmaeops septentrioni s ------6 ------3 11 20 20 - - - 2 ------2 2 cursor - - - - 1 ------1 - - - 2 2 Rhagium bifasciatum ------1 - - 2 2 2 1 1 9 9 Rhagium inquisitor - 3 3 6 3 4 1 2 1 - - 16 2 11 5 4 55 58 Rhagium mordax ------4 1 10 4 4 23 23 Rhagium sycophanta - - - - 1 - - 1 - 3 1 6 5 10 4 3 34 34 Rhamnusium bicolo r ------1 1 1 Necydalinae Necydalis major - - - 1 ------1 1 i scabricorne 1 - 1 ------1 Prionini Prionus coriarius 72 68 140 42 974 3 - - 183 107 2 9 7 1 261 220 1809 1949 Spondylidinae ferus 21 10 31 - 2 1 - - - - - 5 - 7 5 1 21 52 s 6 17 23 2 - - - 8 1 - - 14 - 15 15 27 82 105 Arhopalus syriacus 3 8 11 - 2 ------1 - 1 13 18 35 46 striatum - - - 18 - 2 2 7 - - - 2 - 3 5 1 40 40 castaneum ------16 - 1 15 2 34 34 ------5 - 2 3 - 10 10 Tetropium gabrieli - - - 9 - 1 ------3 14 7 34 34 Saphanini - - Oxypleurus nodieri ------1 1 1 piceus ------1 1 1 Spondylidini buprestoides 3 44 47 2 1 - - 4 2 - - 13 1 4 11 28 66 113

Total individuals 869 2091 2960 618 1748 68 61 229 870 323 9 531 1371 2510 910 944 10193 13153 No. Species 32 42 49 32 37 26 11 18 17 12 5 41 28 49 54 54 107 118

1 3 Journal of Pest Science (2019) 92:281–297 289 in common. Trappings by both lures signifcantly difered traps baited with lure #1 + HV (Table 3). Lure #1 + HV was from those by control traps (Dunn–Nemenyi post hoc test; most attractive for the Lamiinae M. galloprovincialis and #1 vs. #T: P = 0.004; #2 vs. #T: P = 0.0343), which caught M. sutor and the Spondylidinaes and A. 26 species but with a much lower number of specimens (68 striatum, suggesting synergism of the pheromone blend #1 vs. 618 by #1 and 1748 by #2, respectively), and only four by host plant volatiles (Table 4). On the other hand, lure species which were not caught in traps baited with lures #1 #2 also trapped signifcantly more species than lure #2 + N or #2. Three Cerambycinae were signifcantly attracted to (Q1,4 = 4.5; P = 0.040) and was most attractive for the cer- blend #2 (Phymatodes testaceus, alni, and Pyr- ambycines P. testaceus, P. alni, and P. sanguineum, and the rhidium sanguineum; Table 4), while four Lamiinae species Prioninae P. coriarius, suggesting inhibition by addition of were signifcantly attracted by blend #1 (Leiopus nebulo- 2-nonanone (Table 4). sus, , Monochamus galloprovincialis, In 2016, a total of 4412 cerambycids from 60 species were and M. sutor; Table 4). Additionally, signifcant numbers of trapped (Table 3). With a total of 48 species trapped, and a the Lepturinae and the Spondylidinae mean capture of 7.70 ± 0.85 species per site, the combina- were caught in traps baited with lure #1, tion of lure #1 + HV and lure #2 hung side-by-side on the and large numbers of the Prioninae Prionus coriarius were same trap had a signifcantly greater trapping efciency than caught in traps baited with lure #2 (Table 4). lure #2 alone (total species: 28; mean per site: 4.15 ± 0.56; In 2015, a total of 1492 cerambycids in 37 species were Q1,19 = 8.0; P = 0.005), as well as lure #1 + HV alone trapped, the smaller numbers due in part to the reduction (total species: 41; mean per site: 5.25 ± 0.62; Q1,19 = 7.12; in the number of study sites (Table 2). The mean number P = 0.007; Fig. 1a). Although some variations were observed of species trapped per site was roughly the same as in 2014 between sites (see supplementary material for detailed fg- for lure #1 and lure #2 (Fig. 1a). However, the addition of ures), the number of species caught in traps baited with the kairomones resulted in some changes in the patterns of trap combination of the two lures did not difer from the sum of captures across treatments, including both the mean num- the species caught in traps baited with each lure separately ber of species (Q4,4 = 13.9, P = 0.008; Fig. 1a) as well as (total species: 54; mean: 8.30 ± 0.83 species; Q1,19 = 0.059; the mean numbers of specimens caught per site although P = 0.81; Fig. 1a). Subfamilies and tribes were similarly less signifcant (Q4,4 = 4.0; P = 0.045; Fig. 1b). Slight but represented by numbers of species in traps baited with the signifcant diferences in the mean number of trapped spe- lure combination and the sum of the captures in separate cies were noted between lures #1 and #1 + HV when consid- traps baited with either lure #1 + HV or lure #2 (Fig. 2a; ered separately (Q1,4 = 4.0; P = 0.045). That is, traps baited t = −1.41, df = 4; P = 0.23 for subfamilies; t = −1.55, df = 22; with lure #1 + HV, which included α-pinene and ethanol, P = 0.14 for tribes). Captures of beetles in traps baited with caught 18 species, whereas traps baited with lure #1 caught the combination of lures did not difer signifcantly from only 11 species, of which three species were not caught in captures in traps baited with lure #2 (Q1,19 = 1.8; P = 0.18)

Fig. 1 Mean number (± se) of cerambycid species per trap (a) and 2017. Means within a year with diferent letters are signifcantly dif- individuals per trap (b) collected from 2015 to 2017, in traps baited ferent (Friedmann’s Q test followed by Dunn–Nemenyi multiple com- with diferent lures and placed in forests in France from 2014 to parison test, P < 0.05). Lure abbreviations refer to Table 1

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Table 4 Mean (± SE) number of beetles captured per lure and rep- panol; #2: 3-hydroxy-2-hexanone, prionic acid, 2-methylbutan-1-ol, licate for species which showed statistically signifcant overall treat- and anti-2,3-hexanediol in isopropanol; #T: isopropanol; #1 + HV: ment efects (Friedman’s test P < 0.05) in French forests during a blend #1, (-)α-pinene and ethanol in three separate bags; #2 + N: given year from 2014 to 2017. Replicates that contained no speci- blend #2 and 2-nonanone in two separate bags; #(1 + HV) + #2: blend mens of the species in question in any of the treatments were not #1, (-)α-pinene, ethanol and blend #2 in four separate bags; #3: + HV: included in analyses. Exotic species are underlined. Lures: #1: fuscu- blend #1 and blend #2 combined in isopropanol, and (-)α-pinene and mol, fuscumol acetate, monochamol, and geranylacetone in isopro- ethanol in three separate bags Year 2014 2015 2016 2017 Lure #1 #2 #T #1 #1+HV#2#2+N#T#1+HV #2 #(1+HV)+ #2 #(1+HV)+#2 #3+HV Cerambycinae Callidiini Phymatodes testaceu s 0.21±0.09b2 21.58±6.21a0.05±0.05b 0c 0c 106.00±73.00a 61.00±56.00b 0c 0.35±0.17c 16.50±7.01b67.60±31.80a 3.10±0.90b 4.12±0.79a 1 Q2,18=30.5*** Q4,4=17.9** Q2,19=32.6*** Q1,47=3.5* Poeciliumalni 0.22±0.22b8.44±2.45a 0.44±0.44b 0c 0c 191.50±167.50a 35.50±23.50b 0c 0.09±0.09b47.00±12.25a 22.27±7.10a Q2,8=12.1** Q4,4=15.3** Q2,10=14.2** Pyrrhidium sangu ineum 0b 26.25±8.03a0.75±0.75b 0c 0c 23.67±16.60a 4.33±3.38b0c0.38±0.38b35.31±12.31a33.39±14.63a Q2,7=10.8** Q4,2=10.9* Q2,12=15.3**

Xylotrechus stebbing i 0b 1.75±1.11a1.25±0.48a Q2,4=9.9** Lamiinae Acanthocinini 5.83±2.91a0b5.83±2.06a Q2,5=7.2* Leiopus nebulosu s 5.75±3.65a 0b 0.13±0.13b Q2,7=10.8** Acanthoderini Aegomorphus clavipes 10.55±2.99a0b0.18±0.18b 1.09±0.043a 0.09±0.09b1.09±0.32a Q2,10=21.4*** Q2,10=6.2* Monochamini Monochamus 18.00±3.41a0.31±0.24b 0.69±0.29b 7.20±7.20b 27.00±11.57a 0c 0c 0c 17.69±4.70a0.31±0.25b 15.44±5.35a2.68±0.60b3.01±0.52a galloprovinci alis Q2,12=22,4*** Q4,4=18.3** Q2,15=20.8*** Q1,58=4.8* Monochamus suto r 13.60±4.93a0.40±0.24b 0b 1.67±1.67b 8.33±2.33a 0c 0.33±0.33c0c Q2,4=9.3* Q4,2 =9.3* Lepturinae Lepturini Anastrangalia dubi a 1.75±0.48a 0.25±0.25b 0.25±0.25b Q2,3=8.0* Prioninae Prionini Prionus coriarius 4.20±2.06b 97.4±20.13a0.30±0.21b 0c 0c 91.50±88.50a 53.50±52.50b 1.00±1.00c Q2,9=18.9** Q4,4=19.2*** Spondylidinae Asemini Arhopalus ruscus 0b 2.00±1.00a 0.25±0.25b 0b 0b Q4,4=14.4** Asemum striatum 2.57±0.78a 0b 0.29±0.18b 0.40±024b 1.40±0.51a 0c 0c 0c Q2,6=13.1* Q4,4=12.2* 1 Asterisks indicate signifcance level of Friedman’s Q: *P < 0.01; **P < 0.001; ***P < 0.0001 2 Means within species with diferent letters are signifcantly diferent at P < 0.05 following Dunn–Nemenyi post hoc test but was signifcantly greater than captures in traps baited 14 species which were not captured in traps baited with with lure #1 + HV (Q1,19 = 5.0; P = 0.025; Fig. 1b). How- the complete lure combination. There was no signifcant ever, no diferences in captures were observed between the diference in the mean number of individuals trapped per lure combination and the sum of the captures in traps baited site between lures (Q1,19 = 0.888; P = 0.35; Fig. 1b). When with each lure separately (Q1,19 = 0.20; P = 0.66). Only P. collecting dates was considered, the mean number of spe- testaceus was most strongly attracted to the complete com- cies trapped per site and date did not difer signifcantly bination of lures (Table 4). Addition of lure #2 had no efect between the lures (#3 + HV: 2.28 ± 0.15; lure combination: on attraction to #1 + HV for the Lamiinae Leiopus linnei, 2.00 ± 0.17; Q1,128 = 1.4, P = 0.23), nor the mean numbers Aegomorphus clavipes, and M. galloprovincialis. Similarly, of individuals (#3 + HV: 7.85 ± 0.99; lure combination: attraction of the Cerambycinae P. alni, P. sanguineum, and 7.69 ± 0.11; ­Q1,128 = 2.89, P = 0.089). Only two species X. stebbingi to lure #2 was not infuenced by addition of lure showed weak but signifcant treatment efects: the Ceram- #1 + HV (Table 4). bycinae P. testaceus and the Lamiinae M. galloprovincia- In 2017, a total of 1854 cerambycids in 70 species were lis (Table 4). Species captured by subfamilies and tribes of trapped (Table 3). The mean capture of cerambycid species Cerambycidae were also roughly similar between the two per site did not difer between lure #3 + HV, containing all lures (Fig. 2b; t = −0.13, df = 4; P = 0.90 for subfamilies; eight of the cerambycid pheromones, and the combination t = −0.13, df = 22; P = 0.90 for tribes). of lure #1 + HV and lure #2 hung in separate sachets on the same trap (Q1,19 = 0.25; P = 0.67; Fig. 1a). Both attracted Trapping at potential points‑of‑entry similar numbers of 54 species, sharing 40 species in com- mon. Traps baited with the lure combination caught 16 Overall captures included 2960 cerambycids in 49 spe- species which were not captured in traps baited with lure cies (Table 3), with 32 species trapped within the sites and #3 + HV. Conversely, traps baited with lure #3 + HV caught 42 within a radius of 1 km. All but seven species trapped

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Fig. 2 Comparison of the number of species caught per tribe in for- blends in 2016; b captures with the eight-component lure plus host ests in France by traps baited with each multi-lure in 2016 and 2017. volatiles (lure #3 + HV) versus captures with the two four-component a Captures with the two four-component blends (lure #1 + HV and blends (lure #1 + HV and lure #2) hung in separate sachets on the lure #2) hung in separate sachets on the same trap versus sum of the same trap in 2017 captures by separate traps baited with each of the four-component within the sites were also trapped in the vicinity, the spe- the Cerambycinae X. stebbingi, which had been previously cies trapped only within the sites being represented by reported in Europe, was caught in signifcant numbers in all only 1 or 2 specimens. Three exotic Asian species were years, both within potential points-of-entry (54 specimens) captured within potential points-of-entry, including one and in their vicinity (57 specimens), but only in traps with specimen of the Lamiinae Uraecha angusta, and one speci- lures containing blend #2. A Mediterranean species, Poeci- men of the Cerambycinae Xylotrechus altaicus, both being lum fasciatum (Cerambycinae, tribe Callidiini), was also trapped within the Rungis trade market, near Paris, in 2015, captured in 2016 by lure #2 within the Huningue river port by lure #1 + HV and lure #2, respectively. Neither species in Alsace, located much further north. Large numbers of the had ever been recorded before in Europe. A third species, native M. galloprovincialis, the primary vector of pinewood

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Fig. 3 Mean number (± se) of cerambycid species per trap (top graphs) and individuals per trap (bottom graphs) collected from 2015 to 2017, in traps baited with diferent lures and placed at points-of-entry (a, b) or in the vicinity (c, d). Lure abbreviations refer to Table 1 nematode in Europe, were captured in traps baited with lures no diference in the mean number of species captured per containing blend #1 within and in the vicinity of the Medi- site was observed between lure #1 + HV and lure #2 both terranean ports of Fos and Marseille, and a few specimens within the site (Q1,11 = 16.64; P = 0.083; Fig. 3a) and in also were captured each year within the Roissy-Charles de the vicinity (Q1,11 = 11.02; P = 0.27; Fig. 3c). In 2017, Gaulle airport. 21 species were trapped within sites (359 individuals), Large variations in both the number of species, ranging whereas 27 species (828 specimens) were captured in the from 12 to 26 within potential points-of-entry (Fig. 3a) vicinity. Sixteen species were trapped in both the points- and 17–37 in their vicinity (Fig. 3c), and the numbers of of-entry sites and the vicinity, including four in signifcant specimens (Fig. 3b, d) were observed among years, even numbers, the natives M. galloprovincialis (13 individuals though the number of surveyed sites only varied slightly. within sites vs. 28 in the vicinity), P. testaceus (44 vs. 78), In 2015, only 12 species (27 specimens) were trapped and P. coriarius (197 vs. 296) and the exotic X. stebbingi within sites versus 17 species (167 individuals) in the (41 vs. 30). vicinity, with only six species in common. The mean num- ber of cerambycid species captured per point-of-entry did not difer between lures both within site (Q4,10 = 4.537; Overall trapping efciency P = 0.34; Fig. 3a) and in the vicinity (Q4,10 = 6.125; P = 0.19; Fig. 3c), but the composition of the species Figure 4 shows the comparison of the taxonomy of the caught in traps baited with each of the fve lures difered 114 native species caught in traps baited with the multi- considerably, as was observed with baited traps in forests lures during this 4-year period compared with that of the during the same year. In 2016, the captures increased to total of 238 native cerambycid species known to exist in 25 species (412 individuals) within sites and 37 species France (Berger 2012). Captures included 51 species from (1158 individuals) in the vicinity. Similarly, as in 2015, 12 tribes of the Cerambycinae, 29 species in three tribes of

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Fig. 4 Relationships between the native cerambycid fauna of France and the total number of species trapped per tribe (ordered as in Table 2) in traps baited with the multi-lures dur- ing the period 2014–2017

the Lepturinae, 22 species in nine tribes of the Lamiinae, Discussion 10 species in three tribes of the Spondylidinae, and two species in two tribes of the Prioninae. Overall, all known Our data support the predictions of Hanks and Millar (2013) species in 10 of the 41 tribes present in France were caught that multiple pheromones can be deployed in blends, and during our trials, while more than a half of the known spe- paired with host volatiles, to maximize the taxonomic diver- cies in 15 other tribes were captured. sity of cerambycid species attracted while simultaneously minimizing the number of traps and lures required. The choice of the individual components to be mixed in such multi-lure blends has profted from recent advances in our

1 3 294 Journal of Pest Science (2019) 92:281–297 knowledge of cerambycid pheromones; it is clear that there trapping of large numbers of M. galloprovincialis, the pri- is substantial parsimony in pheromone structures within the mary vector of pinewood nematode, in several ports and family worldwide, with a given pheromone often attract- airports and their surroundings should especially be noted. ing related species on diferent continents (Hanks and Mil- For the species trapped in forests in numbers sufcient to lar 2016; Millar and Hanks 2017; Boone et al. 2018). This allow statistical tests, the frst experiments comparing multi- widespread parsimony was largely confrmed during the four component blends composed of four diferent cerambycid years of our study. About 48% of the 238 species native to pheromones showed that many species clearly responded to France were captured, with a rather good generic representa- the blend which included their own known or likely phero- tion of the subfamilies Cerambycinae, Lamiinae, and Spon- mone components, and much less or not at all to the other dylidinae, and additionally the rather surprising capture of blends. For example, only blends containing prionic acid 29 species in three tribes of the Lepturinae, although the two resulted in signifcant captures of males of P. coriarius, known lepturine sex pheromones were not included in our similar to the response of that species reported in an earlier blends (see Millar and Hanks 2017). Only 13 of 41 ceramby- study (Barbour et al. 2011). Several species in the tribe Cal- cid tribes showed no captures in our fight traps, but it must lidiini, including P. testaceus, P. sanguineum, and P. alni, be noted that some of these tribes contain mostly fightless were attracted in large numbers only to the blends containing species (e.g., Dorcadionini). Similar results were observed in 2-methylbutan-1-ol, the known pheromone of P. testaceus, studies testing multi-lure blends on other continents (Hanks and anti-2,3-hexanediol, a component of the pheromone of et al. 2012, 2018; Wickham et al. 2014; Hayes et al. 2016). P. sanguineum (Hanks and Millar 2016). The combination Our results suggest that there are considerable advantages of these two compounds has been shown to attract males of to deploying such “generic” multi-component blends at Phymatodes lengi in North America (Hanks et al. 2012). potential points-of-entry for early detection of exotic species, Similarly, only the blends containing monochamol, a well- by defnition “unknown,” at arrival. For example, during conserved pheromone compound for many Monochamus our study we detected three exotic species within the sites species (Lamiinae: tribe Monochamini) worldwide (Hanks considered as potential points-of-entry. Two of these species and Millar 2016; Boone et al. 2018), attracted two Euro- had never been recorded before in Europe, the Lamiinae pean congeners. The vector of the pinewood nematode in U. angusta, associated with camphor trees (Cinnamomum southwestern Europe, M. galloprovincialis, was attracted camphora) in China (Wang 2014), and the Cerambycinae X. in signifcant numbers in forests, but also in ports and air- altaicus, a pest of larch in Siberia which is on the EPPO A2 ports, and in the Alps, whereas the montane M. sutor was list (OEPP 2005), both having been trapped within the Run- also signifcantly attracted only to blends containing mono- gis trade market, near Paris. The third species also belonged chamol. Two other Lamiinae species, L. nebulosus and A. to the Xylotrechus (tribe ), and both X. altaicus clavipes, were specifcally attracted by blends containing and X. stebbingi were caught in traps baited with blends that monochamol, fuscumol, and fuscumol acetate, probably included anti-2,3-hexanediol, a known pheromone compo- by one or both of the latter two compounds. Similarly, the nent of some other Xylotrechus species in North America Spondylidinae A. striatum was signifcantly attracted by the (e.g., Lacey et al. 2009). Xylotrechus stebbingi, which devel- blend containing fuscumol during 2014, similar to reports ops in broadleaved trees, was already known to have invaded in a recently published study (Millar et al. 2018). southern Europe (Cocquempot and Lindelöw 2010) but our The successive tests of blends with an increasing number trapping studies showed its presence within and near all of pheromone components appeared highly promising for the potential points-of-entry surveyed in Southern France development of attractant-baited traps for operational use. (Fos, Marseille, Port-Vendres, La Turbie) as well as in the The combination of the two four-component blends hung in Mediterranean forests (Gignac, Noves), and also within the same trap was generally similar in trapping efciency, the Toulouse airport and in more northern areas (Orléans measured in numbers of species and individuals caught, to region), far from the Mediterranean region where it was frst the use of two separate traps each baited with one of the introduced. The capture of the Mediterranean P. fasciatum four-component blends. Moreover, attraction of ceramby- in a northern river port also showed that surveys with semio- cids to the eight-component blend did not difer from the chemically baited traps can reveal long-distance movement combination of the two four-component blends hung in of native species aided by human activities. Moreover, the the same trap, with no apparent antagonistic interactions. relative similarity of the cerambycid faunas trapped within However, inhibitory efects among blend components occa- the potential points-of-entry and their vicinity confrmed sionally have been shown in previous experiments, with the observations of Rassati et al. (2018) that woody areas one or more components of a blend reducing or even com- surrounding ports and airports can serve as sources of cer- pletely suppressing attraction of some species. For example, ambycid species which may infest outgoing shipments to Hanks et al. (2012) noted depressed attraction of Neocly- other countries and continents. In particular, the consistent tus acuminatus (Cerambycinae, tribe Clytini) to any lures

1 3 Journal of Pest Science (2019) 92:281–297 295 that contained 3-hydroxy-2-hexanone, and Aegomorphus Europe (e.g., Euplatypus paralellus and Xyleborus ferrug- modestus (Lamiinae, tribe Acanthoderini) was signifcantly ineus; data not shown). more attracted to its pheromone when presented as a single Although the present eight-component pheromone blend, compound rather than as a component of a blend. In another along with ethanol and α-pinene, can be considered a rather experiment, Hanks et al. (2018) showed that fuscumol alone good generic attractant for species in a number of ceramby- attracted more beetles of Tetropium species (Asemini) than cid tribes (e.g., Callidiini, Clytini, Acanthocinini, Acantho- when blended with pheromones of other species. However, derini, Monochamini, Prionini, Asemini), some other tribes in our study during 2017, only two species (P. testaceus and were not attracted at all (e.g., the Lamiinae tribes Agapan- M. galloprovincialis) were trapped in sufciently large num- thiini and Phytoeciini, which are mostly associated with her- bers to test for statistical diferences among lures, and in baceous plants), or showed limited responses. There are sev- neither case was there evidence of strong antagonism. eral possible reasons for this lack of response. First, in our Addition of the generic host plant volatiles α-pinene and trials, we used only a small subset of the known cerambycid ethanol to lures may also have afected captures of ceram- pheromones. Second, there is growing evidence that some bycids. Traps baited with combinations of these compounds cerambycid species may use pheromones that are possibly with cerambycid pheromones have previously revealed con- species-specifc, or may be shared by only a limited number tradictory efects, varying across species (Hanks et al. of close relatives. For example, (E)-2-cis-6,7-epoxynonenal, 2012; Sweeney et al. 2014; Collignon et al. 2016). In some the pheromone of the invasive Asian species bungii, cases, the host plant volatiles have synergized attraction to appears to be species-specifc (Xu et al. 2017). Third, phero- pheromone blends, whereas in others, these compounds mones or likely pheromones have been identifed for less appeared to inhibit attraction to the blends. For example, than 1% of the ~ 35,000 described species of cerambycids, Hanks and Millar (2013) noted that the addition of α-pinene and so it is certain that additional pheromone structures enhanced attraction of species that were conifer specialists, remain to be discovered, and at least some of these are likely whereas ethanol enhanced attraction of hardwood special- to be shared among related taxa. For example, since our tri- ists. Attraction of the Spondylidinae A. striatum to the blend als commenced in 2012, several new and seemingly generic containing fuscumol was enhanced by host plant volatiles, cerambycid pheromone structures have been discovered, as reported earlier (Hanks and Millar 2013). In contrast, the such as 1-(1H-pyrrol-2-yl)-1,2-propanedione, which to date Spondylidinae A. rusticus was attracted only by the same has been found in species from Asia (Zou et al. 2016), North pheromone blend combined with host plant volatiles, sug- America (Diesel et al. 2017; Millar et al. 2018), and South gesting a critical synergism. In our trials, the only notice- America (Silva et al. 2017), and 3-methylthiopropan-1-ol, able efect resulting from addition of α-pinene and ethanol found in species in both South America (Silva et al. 2017) was a fourfold increase in captures of the hardwood spe- and North America (JGM and LMH, unpub. data). Finally, cialist P. testaceus in 2016 by the combination of lures it is possible that some species of cerambycids may not use including these volatiles. Sweeney et al. (2014) observed attractant pheromones at all. However, the vast majority of a similar increase in the mean catch of this species in the the species which have been specifcally targeted for phero- Russian Far East when ethanol was combined with racemic mone identifcation to date have indeed been shown to use 3-hydroxyhexane-2-one. pheromones. Thus, traps baited with judiciously chosen Our results suggest that an optimal detection strategy blends of pheromones should be useful tools for detection would be to use pheromone blends in combination with the of at least a signifcant proportion of exotic species that are generic host plant volatiles ethanol and α-pinene, although potentially invasive. reduced attraction caused by inhibition phenomena could potentially be a problem when trapping exotic species at potential points-of-entry where the population density is Author contributions usually very low (Hanks et al. 2012). However, as long as a species’ inhibition is not total, such traps could still con- AR, JT, OD, JGM and LMH conceived and designed the stitute an acceptable compromise, because the presence of research. CC formulated the chemical blends, and JGM pro- ethanol and α-pinene would enhance the simultaneous cap- vided some of the chemicals. JT, OD, AB, MJ conducted ture of exotic bark beetles and other wood-boring beetles. In experiments. OD and AR identifed the specimens. AR did our experiments, few bark beetles (less than 50 individuals) the statistical analyses. JT and AR wrote the paper, and all were caught in traps only baited with the cerambycid blends authors assisted with editing and proofng. within potential points-of-entry. In contrast, addition of these two volatiles to the lure blends resulted in the capture Acknowledgements Jean-Luc Flot, Frédéric Delport and all staf of the French Forest Health Department with the French Ministry of of several hundred specimens of bark and ambrosia beetles, Agriculture are gratefully acknowledged for their support. We are also including several species not yet known to be established in

1 3 296 Journal of Pest Science (2019) 92:281–297 grateful to the following colleagues for technical assistance during feld Cocquempot C, Lindelöw Å (2010) Longhorn beetles (Coleoptera, trials: Christian Blazy, Béatrice Courtial, Matthieu Le Floc’h, Philippe Cerambycidae). BioRisk 4:193–218 Lorme, Emmanuelle Magnoux, Marie Millier, Julien Papaïx, Régis Collignon RM, Swift IP, Zou Y, McElfresh JS, Hanks LM, Millar JG Phélut, Patrick Pineau, Christelle Robinet, Lionel Roques, Olivier (2016) The infuence of host plant volatiles on the attraction of Roques, Géraldine Roux. We thank Joël Giraud, Patrick Vigne, and longhorn beetles to pheromones. J Chem Ecol 42:215–229 the municipality of L’Argentière-la-Bessée for logistic assistance, and Diesel NM, Zou Y, Johnson TD, Diesel DA, Millar JG, Mongold-Diers Laurent Blanchard and the National Forestry Ofce (Agence Territo- JA, Hanks LM (2017) The rare North American cerambycid beetle riale des Hautes-Alpes) for the authorization of trapping in the “Les sexnotatus shares a novel pyrrole pheromone component Deslioures” natural reserve of the Fournel forest. This research was with species in Asia and South America. J Chem Ecol 43:739–744 supported by successive grants from the French Ministry of Agriculture Graham EE, Mitchell RF, Reagel PF, Barbour JD, Millar JG, Hanks (Projects PORTRAP I-2015/045, PORTRAP II-2016/098, and POR- LM (2010) Treating panel traps with a fuoropolymer enhances TRAP III-2017/276 “Test de l’efcacité de pièges génériques multi- their efciency in capturing cerambycid beetles. J Econ Entomol composés pour la détection précoce d’insectes exotiques xylophages 103:641–647 dans les sites potentiels d’entrée sur le territoire national”) and by Handley K, Hough-Goldstein J, Hanks LM, Millar JG, D’Amico V the EUPHRESCO MULTITRAP project (“Multi-lure and multi-trap (2015) Species richness and phenology of cerambycid beetles in surveillance for invasive tree pests”). urban forest fragments of northern Delaware. Ann Entomol Soc Am 108:251–262 Compliance with ethical standards Hanks LM, Millar JG (2013) Field bioassays of cerambycid phero- mones reveal widespread parsimony of pheromone structures, enhancement by host plant volatiles, and antagonism by compo- Conflict of interest All authors declare they have no confict of inter- nents from heterospecifcs. Chemoecology 23:21–44 est. Hanks LM, Millar JG (2016) Sex and aggregation-sex pheromones of cerambycid beetles: basic science and practical applications. J Ethical standard All authorization to carry out experiments in ports Chem Ecol 42:631–654 and forests has been obtained. Experiments in forests were carried out Hanks LM, Millar JG, Mongold-Diers JA, Wong JCH, Meier LR et al in state-owned forests, managed by the French Ministry of Agriculture (2012) Using blends of cerambycid beetle pheromones and host which gave the grants. All applicable international, national, and/or plant volatiles to simultaneously attract a diversity of cerambycid institutional guidelines for the care and use of were followed. species. Can J For Res 42:1050–1059 This article does not contain any studies with human participants per- Hanks LM, Mongold-Diers JA, Atkinson TH, Fierke MK, Ginzel MD formed by any of the authors. et al (2018) Blends of pheromones, with and without host plant volatiles, can attract multiple species of cerambycid beetles simul- taneously. J Econ Entomol 111:716–724 Hayes RA, Grifths MW, Nahrung HF, Arnold PA, Hanks LM, Mil- lar JG (2016) Optimizing generic cerambycid pheromone lures References for Australian biosecurity and biodiversity monitoring. J Econ Entomol 109:1741–1749 Allison JD, Johnson CW, Meeker JR, Strom BL, Butler SM (2011) Lacey ES, Ginzel MD, Millar JG, Hanks LM (2004) Male-produced Efect of aerosol surface lubricants on the abundance and richness aggregation pheromone of the cerambycid beetle of selected forest insects captured in multiple-funnel and panel acuminatus acuminatus. J Chem Ecol 30:1493–1507 traps. J Econ Entomol 104:1258–1264 Lacey ES, Millar JG, Moreira JA, Hanks LM (2009) Male-produced Allison JD, Bhandari BD, McKenney JL, Millar HG (2014) Design aggregation pheromones of the cerambycid beetles Xylotrechus factors that infuence the performance of fight intercept traps for colonus and fulminans. J Chem Ecol 35:733–740 capture of longhorned beetles (Coleoptera: Cerambycidae) from Liebhold A, Brockerhof E, Garrett L, Parke J, Britton K (2012) Live the subfamilies Lamiinae and Cerambycinae. Plos ONE 9:e93203 plant imports: the major pathway for the forest insect and patho- Barbour JD, Millar JG, Rodstein J, Ray AM, Alton DG et al (2011) gen invasions of the US. Front Ecol Environ 10:135–143 Synthetic 3,5-dimethyldodecanoic acid serves as a general attract- Meier LR, Zou Y, Millar JG, Mongold-Diers JA, Hanks LM (2016) ant for multiple species of Prionus (Coleoptera: Cerambycidae). Synergism between enantiomers creates species-specifc phero- Ann Entomol Soc Am 104:588–593 mone blends and minimizes cross-attraction for two species of Bashford R (2008) The development of a port surrounds trapping sys- cerambycid beetles. J Chem Ecol 42:1181–1192 tem for the detection of exotic forest insect pests in Australia. Millar JG, Hanks LM (2017) Chemical ecology of cerambycids. In: In: Oteng-Amoako AA (ed) New advances and contribution to Wang Q (ed) Cerambycidae of the world: biology and pest man- forestry research. InTech, Rijeka, pp 85–100 agement. CRC Press/Taylor & Francis, Boca Raton, pp 161–208 Bense U (1995) Longhorn beetles—illustrated key to Cerambycidae Millar JG, Richards AB, Halloran S, Zou Y, Boyd E, Quigley KN, and of Europe. Margraf Verlag, Weikersheim Hanks LM (2018) Pheromone identifcation by proxy: identifca- Berger P (2012) Coléptères Cerambycidae de la faune de France conti- tion of aggregation-sex pheromones of North American ceramby- nentale et de Corse. Association Roussillonnaise d’Entomologie, cid beetles as a strategy to identify pheromones of invasive Asian Perpigan congeners. J Pest Sci. https://doi.org/10.1007/s1034​ 0-018-0962-4​ Boone CK, Sweeney J, Silk P, Webster RP, Stephen F et al (2018) Mitchell RF, Graham EE, Wong JC, Reagel PF, Striman BL et al (2011) Monochamus species from diferent continents can be efectively Fuscumol and fuscumol acetate are general attractants for many detected with the same trapping protocol. J Pest Sci. https​://doi. species of cerambycid beetles in the subfamily Lamiinae. Entomol org/10.1007/s1034​0-018-0954-4 Exp Appl 141:71–77 Brockerhof EG, Jones DC, Kimberley MO, Suckling DM, Donald- Mitchell RF, Reagel PF, Wong JCH, Meier LR, Silva WD et al (2015) son T (2006) Nationwide survey for invasive wood-boring and Cerambycid beetle species with similar pheromones are segre- bark beetles (Coleoptera) using traps with pheromones and kair- gated by phenology and minor pheromone components. J Chem omones. For Ecol Manag 228:234–240 Ecol 41:431–440

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OEPP (2005) Data sheets on quarantine pests, Xylotrechus altaicus. Sweeney JD, Silk PJ, Grebennikov V (2014) Efcacy of semiochemi- EPPO Bull 35:406–408 cal-baited traps for detection of longhorn beetles (Coleoptera: Cer- Rabaglia RJ, Duerr D, Acciavatti RE, Ragenovich I (2008) Early detec- ambycidae) in the Russian Far East. Eur J Entomol 111:397–406 tion and rapid response for non-native bark and ambrosia beetles. Wang ZC (2014) Monographia of original colored longicorn beetles US Department of Agriculture Forest Service, Forest Health Pro- of China. Scientifc and Technical Documentation Press, Beijing tection, Washington, DC (in Chinese) Rassati D, Petrucco Tofolo E, Roques A, Battisti A, Faccoli M (2014) Webster RP, Alderson CA, Webster VL, Hughes CC, Sweeney JD Trapping wood-boring beetles in Italian ports: a pilot study. J (2016) Further contributions to the (Coleoptera, Pest Sci 87:61–69 Cerambycidae) fauna of New Brunswick and Nova Scotia, Can- Rassati D, Faccoli M, Petrucco Tofolo E, Battisti A, Marini L (2015) ada. ZooKeys 552:109–122 Improving the early detection of alien wood-boring beetles in Wickham JD, Harrison RD, Lu W, Guo Z, Millar JG et al (2014) ports and surrounding forests. J Appl Ecol 52:50–58 Generic lures attract cerambycid beetles in a tropical montane Rassati D, Haack RA, Knížek M, Faccoli M (2018) National trade rainforest in southern China. J Econ Entomol 107:259–267 can drive range expansion of bark- and wood-boring beetles. J Wong JCH, Mitchell RF, Striman BL, Millar JG, Hanks LM (2012) Econom Entomol 111:260–268 Blending synthetic pheromones of cerambycid beetles to develop Roques A (2010) Alien forest insects in a warmer world and a glo- trap lures that simultaneously attract multiple species. J Econ balized economy: impacts of changes in trade, tourism and climate Entomol 105:906–915 on forest biosecurity. NZ J For 40:77–94 Xu T, Yasui H, Teale SA, Fujiwara-Tsujii N, Wickham JD et al (2017) Seebens H, Blackburn TM, Dyer EE, Genovesi P, Hulme PE et al Identifcation of a male-produced sex-aggregation pheromone for a (2018) Global rise in emerging alien species results from highly invasive cerambycid beetle, Aromia bungii. Sci Rep 7:7330 increased accessibility of new source pools. PNAS. https​://doi. Zou Y, Rutledge CE, Nakamuta K, Maier CT, Hanks LM et al (2016) org/10.1073/pnas.17194​29115​ Identifcation of a pheromone component and a critical synergist Silva WD, Zou Y, Bento JMS, Hanks LM, Millar JG (2017) Aggrega- for the invasive beetle rufpenne (Coleoptera: Cer- tion-sex pheromones and likely pheromones of 11 South Ameri- ambycidae). Environ Entomol 45:216–222 can cerambycid beetles, and partitioning of pheromone channels. Front Ecol Evol 5:101

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