J. ENTOMOL. SOc. BRIT. COLUMBIA 117, DECEMBER 2020 { 5 Ra pe iat 2

Journal of the

Entomological Society of British Cotléentia”

Volume 117 December 2020 ISSN#0071-0733

Directors of the Entomological Society of British Columbia 2020—2021.................. 2

The balsam bark weevil, Pissodes striatulus (Coleoptera: Curculionidae): life history

and. oecutrence.in. southern: british GC Ohana bias yi s95 a) ede. caw nee ee ea eulid 3

Beetles in the city: ground beetles (Coleoptera: Carabidae) in Coquitlam, British

Columbia as indicaters.of human-disturbanee 23.5.0 a GAN ER a eek 20

Effects of trail pheromone purity, dose, and type of placement on recruiting European

fire ants, Myrmica rubra. Ae BOOC AIS bce in Pts Ver ilaglnw ache ates veer tees es at

Geographic range and seasonal occurrence in British Columbia of two exotic ambrosia

beetles as determined by semiochemical-based trapping .................. see ceeeee eee e ence 42

Andrena (Melandrena) cyanura Cockerell (Hymenoptera: Apoidea, Andrenidae), a valid

North America SO e1ee oles ines sussuinklnaas +e niaincoenleeeyees te Te ee ee, 49

SCIENTIFIC NOTES

First record of the Palearctic seed bug Metopoplax fuscinervis Stal (Hemiptera:

OxyCareniGae) Th GRIT A SICA sie se sevecsny 050-9 4c5 oes 9 nuee OR Pe ene umm aio aie 60

NATURAL HISTORY AND OBSERVATIONS

Plecoptera from the’ Crooked River, British Columbia °. o.oo. ace ogg re ees anes sh eee ean 64

New distribution records and range extensions of mosquitoes (Diptera: Culicidae) in

British Columbia the Wokcon Territory os... ss0csessannrsinesahetedesnae pwassanaehe borden 69

OBITUARY: Leland Medley Humble (3 November 1951 - 4 August 2020)

NOTICE TO CON Tae 1 ey ask oe OR ak as ba i BS Inside Back Cover

J. ENTOMOL. SOc. BRIT. COLUMBIA 117, DECEMBER 2020

DIRECTORS OF THE ENTOMOLOGICAL SOCIETY

OF BRITISH COLUMBIA FOR 2020-2021

President:

Wim van Herk (president@entsocbc.ca)

Agriculture and Agri-Food Canada, Agassiz

Ist Vice President:

Chandra Moffat

Agriculture and Agri-Food Canada, Summerland

2nd Vice-President:

Lorraine Maclauchlan :

BC Ministry of Forests, Lands, Natural Resource Operations and Rural Development, Kamloops

Past-President:

Tammy McMullen

Simon Fraser University, Burnaby

Treasurer:

Markus Clodius (membership@entsocbc.ca)

Agriculture and Agri-Food Canada, Agassiz

Secretary:

Rob Higgins (secretary@entsocbc.ca)

Thompson Rivers University, Williams Lake

Directors:

Joyce Leung Dan Peach

Simon Fraser University, Burnaby University of British Columbia, Vancouver

Graduate Student Representative:

Asim Renyard

Simon Fraser University, Burnaby

Regional Director of National Society: ao, ee

Brian Van Hezewijk (boreus@entsocbc.ca)

Canadian Forest Service, Victoria Canadian Food Inspection Agency,

Kelowna

Web Editor: Assitant Editor, Boreus:

Brian Muselle (webmaster@entsocbc.ca) Elton Ko

University of British Columbia, Okanagan Simon Fraser University, Burnaby

Society homepage: Journal homepage:

http://entsocbc.ca http://journal.entsocbc.ca

Editorial Committee of the Journal of the Entomological Society of British Columbia:

Editorial Board: Joel Gibson, Dezene Huber,

Bob Lalonde, Bo Staffan Lindgren, Lorraine

Maclauchlan, Robert McGregor, Steve

Perlman, Lisa Poirier, Marla Schwarzfeld

Editor-in-Chief:

Katherine Bleiker (journal@entsocbc.ca)

Canadian Forest Service, Victoria

Copy Editor: Monique Keiran Technical Editor: Jesse Rogerson

J. ENTOMOL. Soc. BRIT. COLUMBIA 117, DECEMBER 2020 3

The balsam bark weevil, Pissodes striatulus (Coleoptera:

Curculionidae): life history and occurrence in southern British

Columbia

L.E. MACLAUCHLAN! AND J.E. BROOKS?

ABSTRACT

Subalpine fir (Abies lasiocarpa (Pinaceae)) forests in British Columbia (B.C.)

are increasingly climate-stressed and vulnerable to pest damage. Following a

drought in southern B.C., the balsam bark weevil, Pissodes_ striatulus

(Coleoptera: Curculionidae), was observed attacking and killing mature

subalpine fir trees. This study documents P. striatulus as a tree-killing insect,

often associated with western balsam bark beetle (Coleoptera: Curculionidae),

which is considered the most destructive insect pest of subalpine fir. In B.C.,

this weevil displays a one-year life history, overwintering as late-instar larvae in

the bark and as newly emerged or older adults in the duff at the base of attacked

trees. Attacked trees are difficult to identify until the tree becomes chlorotic and

dies. Larvae may excavate diagnostic chip cocoons in the sapwood before

pupating, but most complete their development in the phloem where their

galleries quickly become obscured by woodborer activity and other insects.

Pissodes striatulus was found at 71% of sites surveyed, and 19% of trees

sampled were killed by the weevil acting as the primary invader. The weevil

uses downed trees, slash, and susceptible live trees, is long lived, and can switch

from primary to secondary attacker, demonstrating its capacity to adapt to

available and changing conditions.

Key words: balsam bark weevil, Pissodes striatulus, subalpine fir, climate

stress

INTRODUCTION

Over the past two decades, subalpine fir Abies lasiocarpa (Hook.) Nutt. (Pinaceae)

mortality in British Columbia (B.C.) has increased due to insect attack, root disease, and

climatic stress (Maclauchlan 2016). Much of this mortality is attributed to the western

balsam bark beetle (WBBB), Dryocoetes confusus Swaine (Coleoptera: Curculionidae),

largely considered the most destructive insect pest of subalpine fir and causing scattered

mortality over large areas of high-elevation forests (Furniss and Carolin 1977; Stock

1991; Garbutt 1992; McMillin et al. 2003; Lalande et a/. 2020). Although the primary

mortality agent may be WBBB, there is little ground survey information on the incidence

and impact of WBBB and other damaging agents in these sensitive and often remote

high-elevation forests.

Subalpine fir ecosystems are threatened by climate extremes, pests, and increased

harvesting (Reich et al. 2016; Lalande et al. 2020). These are extremely valuable forests

due to their inherent hydrologic contribution (Winkler et al. 2017), carbon sequestration,

and habitat attributes. Subalpine fir grows well at high elevations, from 600 to 2 250

metres, throughout most of the B.C. interior (Parish and Thomson 1994). In the

mountains and plateaus of interior B.C., subalpine fir is often associated with spruce

(Pinaceae) and is a major component of the interior high-elevation forests from Yukon,

Canada, to Arizona, United States of America. Cool summers, cold winters, and a deep

1 Ministry of Forests, Lands and Natural Resource Operations and Rural Development, 441 Columbia

Street, Kamloops, B.C. V2C 2T3; Lorraine.maclauchlan@gov.bc.ca

2 Forest Health Management, 466 Central Ave., Gibsons, B.C. VON 1V1

4 J. ENTOMOL. SOc. BRIT. COLUMBIA 117, DECEMBER 2020

snowpack are important in determining where subalpine fir grows well (Parish and

Thomson 1994).

As more non-native insects establish and expand in B.C. and climate change

increases in intensity, many forest insects and fungal pathogens, both native and non-

native, are expected to expand their ranges northwards and to higher elevations and, with

this expansion, bring more severe impacts to newly invaded areas (Bentz et al. 2010;

Woods et al. 2010, 2017; Haughian et al. 2012). The expansion of mountain pine beetle,

Dendroctonus ponderosae Hopkins (Coleoptera: Curculionidae), well beyond its historic

range into northern B.C., Yukon, and Northwest Territories (Safranyik et al. 2010),

coupled with the beetle’s extensive attack in young lodgepole pine (Maclauchlan ef al.

2015), is a well-documented example of a native insect responding to the effects of

climate change on both insect and host.

Another recent example of insect range expansion in B.C. is that of the balsam

woolly adelgid, Adelges piceae (Ratzeburg) (Hemiptera: Adelgidae), an introduced insect

from Europe. It was thought to be contained in a provincial quarantine zone on the B.C.

coast until recent surveys (Zilahi-Balogh et al. 2016; Maclauchlan and Buxton 2018)

confirmed its presence outside the pre-2014 quarantine zone, affecting subalpine fir

throughout the southern interior and as far north as Horsefly in the B.C. Cariboo Region.

While conducting studies on WBBB in subalpine fir forests (Maclauchlan and

Buxton 2016, 2017; Maclauchlan 2020), a weevil was observed attacking and killing

live, mature subalpine fir trees, acting as a primary invader, much like tree-killing bark

beetle species. The weevil was identified as the balsam bark weevil, Pissodes striatulus

(Fabricius) (Coleoptera; Curculionidae) (Randall 1838; O’Brien and Thompson 1986).

Most published records of this weevil are from eastern Canada, primarily Ontario,

Quebec, and New Brunswick, and the northeastern United States of America, where

reports have noted it infesting balsam fir, Abies balsamea (L.) Mill. (Pinaceae) that has

been severely defoliated or killed by eastern spruce budworm, Choristoneura fumiferana

(Clemens) (Lepidoptera: Tortricidae) (Swaine et al. 1924; Craighead 1950; Belyea

1952a). In B.C., there are very few published records for P. striatulus, and it has typically

been observed in association with WBBB. Little is known of the life history, host

selection parameters, and habits of this weevil in B.C.

Swaine et al. (1924) considered P. striatulus to be the most aggressive of the insects

that attacked dead and dying trees following eastern spruce budworm defoliation.

According to his studies, the larvae were reported never to develop to maturity unless the

tree was almost dead, and two or three successive attacks could be made on the same tree

before the tree’s resistance was low enough that weevil larvae could survive and mature

to adulthood. Most observations from eastern forests describe P. striatulus as essentially

a secondary insect (Craighead 1950). Secondary insects usually select hosts that have

impaired defenses and avoid vigourous trees, whereas primary invaders attack and kill

apparently vigourous trees through pheromone-mediated mass attacks. Early

observations on the seasonal history of this weevil differ somewhat depending upon

location (Swaine et al. 1924; Belyea 1952a, 1952b), suggesting it has a plastic life history

and is able to adapt to local and seasonal climate. Belyea (1952a) observed full-grown

larvae and pupae under the bark of infested trees in the Lake Nipigon area of Ontario,

Canada, in late June, with adults emerging from late June through the middle of August,

and the majority emerging in the last three weeks of July. They estimated development

time to be just less than 12 months from egg to emergent adult in this area. Swaine et al.

(1924), however, suggested a two-year life cycle, with the insects overwintering as adults

and emerging from the duff in the spring to mate and lay their eggs. All studies found that

the weevils preferred to oviposit on the lower fifth to lower half of their balsam fir host.

Other Pissodes species have similar host selection parameters to P. striatulus.

Pissodes nemorensis Germar and Pissodes schwarzi Hopk., are attracted to boles, slash,

and root collars of weakened, stressed, or dying trees (Finnegan 1958; United States

Department of Agriculture Forest Service 1985; Atkinson ef al. 1988; Maclauchlan et al.

J. ENTOMOL. SOc. BRIT. COLUMBIA 117, DECEMBER 2020 5

1993). Host selection by P nemorensis and P. schwarzi has been shown to be

pheromone mediated (Fontaine and Foltz 1982; Maclauchlan ef al. 1993), with P.

nemorensis producing grandisol (cis-2-isopropenyl-l-methy1cyclobutaneethanol) and its

corresponding aldehyde, grandisal, which act together as aggregation pheromones (Booth

et al. 1983; Phillips et al. 1984; Phillips and Lanier 1986). P. striatulus may also use

these pheromones and the smell of stressed trees to attract mates and aggregate on

potential host trees.

The objectives of this study were to describe the occurrence, life history, and host

preference of P. striatulus in susceptible stands of subalpine fir in southern British

Columbia. Additionally, we aimed to determine the prevalence of P. striatulus attack in

low-elevation, climate-stressed subalpine fir stands and its interaction with WBBB.

METHODS

Life history sampling and field observations. Three field sites were selected to

study the life history of P. striatulus in southern B.C. A site located off the Spahats Creek

Forest Service Road (51° 46' 25.18" N, 119° 45' 26.77" W; elevation: 1 600 m) northeast

of Clearwater, B.C. in the Engelmann—Spruce—Subalpine Fir—Wet—Cold biogeoclimatic

zone (ESSFwc) (Lloyd et al. 1990; Meidinger and Pojar 1991), and predominantly

subalpine fir, was selected to collect observational data during the summers of 2015—

2016. The other two sites, Watching Creek (50° 54’ 23.64" N, 120° 26' 32.78" W;

elevation: 1 375 m) and Antler Road (50° 52' 54.12" N, 120° 24' 39.96" W; elevation:

1 260 m) were located about 30 km northwest of Kamloops, B.C., on the west side of the

North Thompson River. Both sites are situated in the Montane—Spruce—Dry—Mild

biogeoclimatic subzone (MSdm). The Watching Creek site is composed of spruce,

Douglas-fir, and subalpine fir. The Antler Road site is a mix of spruce and subalpine fir.

From 2015 through 2017, both the Watching Creek and Antler Road sites had a high

population of P. striatulus, and the sites were accessible throughout most of the year.

In 2016 through 2018, a total of 15 live, green subalpine fir trees that were newly

mass attacked by P. striatulus at the Watching Creek and Antler Road sites were selected

for twice-weekly life-stage sampling and observation (March—November). During field

sampling, a ladder was used to access higher portions of the trees to collect bark samples,

which were approximately 20 cm x 20 cm. Larvae and other life stages that were easily

visible when the bark was removed were collected and placed into vials containing 70%

ethanol (EtOH). Each remaining bark sample was placed in a sealable bag, labelled, and

brought into the laboratory, where it was dissected to expose remaining life stages within

one day of collection. All specimens were preserved in 70% EtOH for future

measurement. Ten live adult weevils were collected and sent to L. Humble, Canadian

Forest Service, Victoria, B.C., Canada, to confirm species identification. The following

information was recorded at each field sampling: life stage(s) present; gallery

description; timing of attack and oviposition; presence—absence of chip cocoons or exit

holes; timing of adult emergence; and, bole and foliar symptoms of attacked trees. Adults

collected during field sampling were frozen until measured. All life stage and field

observations were compiled along a timeline to produce a lifetable for P striatulus in

southern B.C.

A Hobo™ U23-003 Pro v2 Temperature Data Logger, with 2 exterior sensors and a

Hobo™ RSI Solar Radiation Shield (Onset Computer Corporation, Bourne,

Massachussetts, United States of America; https://www.onsetcomp.com/) were set up at

the Watching Creek and Antler Road sites to monitor daily mimimum and maximum

temperatures over a full year (January-December 2017). One temperature sensor was

placed at ground level, and the second sensor was placed at 2.5 m on the north side of a

tree situated inside the stand. Temperature records were downloaded to a laptop computer

every two to three weeks. At the Watching Creek site, four 8-funnel Lindgren multiple-

funnel traps (Lindgren 1983) were placed at 25-metre intervals throughout the stand. Two

6 J. ENTOMOL. Soc. BRIT. COLUMBIA 117, DECEMBER 2020

traps were baited with (+)-exo-brevicomin to monitor WBBB flight times, and two >

were baited with grandisal and grandisol (supplied by Synergy Semiochemicals Corp.,

Delta, British Columbia, Canada), to monitor the onset of P. striatulus flight. The traps

were checked when life-stage sampling was conducted at the sites. Average daily

maximum and minimum temperatures were compared to trap catches and life stage

sampling data.

In addition to the periodic sampling of trees, five additional live, green subalpine fir

trees mass attacked by P. striatulus at the Watching Creek and Antler Road sites were

felled to assess attack over the entire length of the trees, to collect life stages, and to

quantify successful weevil emergence (Table 1). Each time an emergence hole was

counted, an X was marked over the hole in indelible marker to ensure a single count.

Felled trees were cut into one-metre sections, with the cardinal direction marked and

labelled according to position in the bole, and then transported to the laboratory, where

the final count of emergence holes was done. The cut ends of all sample sections were

sealed with paraffin wax to prevent desiccation; each section was covered in mesh

screening and placed in a 20 °C environment chamber to allow any further emergence.

The diameter of each one-metre tree section was measured and the bark surface area

calculated. The length and position of each section was converted to reflect height on

individual trees. The diameter of a sub-sample of exit holes was measured. The data were

converted to the number of emergence holes per square metre of bark surface, and the

frequency distribution of emergence density by tree height was calculated.

Table 1. List of subalpine fir trees felled to assess emergence and extent of attack on trees,

noting year of attack, date felled, site (Watching Creek: 50° 54' 23.64" N, 120° 26' 32.78" W;

elevation: 1 375 m; Antler Road: 50° 52' 54.12" N, 120° 24' 39.96" W; elevation: 1 260 m),

and dates each tree was field sampled.

Attack year Date felled Site Field sampling dates

2015 16 Sep. 2016 Watching Creek Sep. — 1 Nov. 2016

2016 16 Nov. 2016 Watching Creek No field sampling

2015 26 Apr. 2017 Watching Creek 6 Jun. — 6 Sep. 2016

2017 18 Jun. 2018 Antler Road 5 Jul. — 22 Nov. 2017

2017 24 Aug. 2018 Antler Road 15 May — 24 Aug. 2018

4 Observed undergoing mass attack by P. striatulus on 5 July 2017.

The head-capsule widths of each collected larva were measured at the widest point

using a Meiji binocular microscope (Meiji Techno, San Jose, California, United States of

America) equipped with a micrometer to determine the number and size range of larval

instars (Stark and Wood 1964; Langor and Williams 1998; Logan et al. 1998; Panzavolta

2007). The head-capsule width data were analyzed using the Hcap program developed by

Logan et al. (1998). Adult weevils collected during field or laboratory sampling were

measured at their widest and longest points to determine size range.

Eleven longterm installations previously established to monitor WBBB attack and

stand succession in subalpine fir forests in southern B.C. (Maclauchlan and Buxton 2017;

Maclauchlan 2020) had occasional records of P. striatulus colonising trees within these

plots. Plots were located in three ESSF subzones: three in the ESSFxc (very dry, cold);

six in the ESSFwe (wet, cold); and two in the ESSFmw (moist, warm) (Lloyd ef al. 1990;

Meidinger and Pojar 1991). The weevil was observed colonising mature subalpine fir

J. ENTOMOL. Soc. BRIT. COLUMBIA 117, DECEMBER 2020 t

alone and in association with WBBB. We summarised all records of P. striatulus

from these 11 plots.

Field surveys. The objective of the field survey was to ascertain presence or absence

of P. striatulus in randomly selected low- to mid-elevation subalpine fir stands by

examining recently dead subalpine fir trees. Using Aerial Overview Survey spatial data

(Ministry of Forests, Lands, Natural Resource Operations and Rural Development 2016),

low- to mid-elevation stands containing subalpine fir and current WBBB attack (red

trees) were identified. From the air, red subalpine fir is typically labelled as mortality

caused by WBBB. Geo-referenced PDF maps were created for candidate areas, loaded

onto a tablet. and using Avenza Systems Inc.® (2017; Toronto, Ontario, Canada) were

located in the field. Survey sites were chosen based upon road access and visible red or

fading subalpine fir near mapped areas of 2016 WBBB. Elevation and GPS coordinates

were recorded for each site that was surveyed. Recently dead subalpine firs (displaying

chlorotic to bright red foliage) were assessed for P. striatulus and other mortality factors.

Occasionally, if older attack (displaying dull red foliage) or green trees undergoing

current attack were present, they were also assessed. The number of trees assessed in

each stand varied, based on the abundance of recently killed trees in the stand. Each tree

was thoroughly checked by first examining the outer bark for exit holes and signs of

Oviposition or resin, and then peeling back the bark to look for weevil galleries, chip

cocoons, and life stages. The foliage colour was recorded as green or chlorotic (new

attack), bright red (prior year attack) and dull red (older attack). Western balsam bark

beetle, woodborers, and any other insects or pathogens found under the bark were noted.

All P. striatulus life stages were collected, labelled, and stored in 70% alcohol. For each

sampled tree, existence of evidence of only P. striatulus attack, only WBBB attack, or

whether both species were observed was recorded.

RESULTS

Life history sampling and field observations. The 10 weevil specimens sent to L.

Humble were confirmed as P. striatulus, the balsam bark weevil (pers. comm.). The life

table shown in Figure 1 was constructed by combining all field observations and life-

stage sampling gathered in 2015 through 2017. On 2 July 2015, at the Spahats Creek site,

numerous adult P. striatulus were observed mating and ovipositing on the freshly cut

surface of a subalpine fir stump at the phloem—cambium interface and on the boles of

standing live subalpine fir. Oviposition was distinguished by small feeding punctures

made by the weevil in the outer bark, where it had laid one to several eggs and then had

capped the puncture with a frass plug. At the oviposition site, a droplet of cloudy, red

pitch (Fig. 2) usually was present, and the tree characteristically produced streams of

resin. In late September, the bark was peeled from the attacked stumps, and the galleries

originating from the cut surface were clearly visible radiating downwards to the root

collar (Fig. 2). The larvae created straight or sometimes winding serpentine-like galleries

downwards from the point of oviposition; as the larvae grew, they often veered off at

right angles and mined around the circumference of the tree. By 25 September, larvae

were large, late instar, and presumably in the overwintering stage. At that time, P.

striatulus larvae (same life stage as in the stump) were also found in standing subalpine

fir attacked by WBBB in 2014. Weevil attack occurred on the lower bole where there was

available phloem not used by WBBB.

In October 2015, the Watching Creek site was first located by field-checking a

fading subalpine fir that displayed a slightly different fade pattern than is normally

associated with trees attacked and killed by WBBB, which typically fade to a bright red

the summer following attack. This tree had dull red foliage with remnants of green on

some branch tips (Fig. 2). Under the bark, galleries around the entire circumference of

the tree, like those at the Spahats Creek site, and late-instar weevil larvae were found,

indicating that attack had likely occurred that summer. The outer bark showed no sign of

8 J. ENTOMOL. SOC. BRIT. COLUMBIA 117, DECEMBER 2020

attack by WBBB, nor was frass or sawdust present around the bole, as is usually the

case with WBBB attack.

Observed

exit holes

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Sioa Sa

Omnacecnncrermaaisepasecrnncewsenaees: stnnsasnennsronannshanasenannannnnas

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ae eer a ea ae er ERT aT NTC

Figure 1. Timing of behaviour and life stages of P. striatulus from first to last observation

(data from 2015-2017). Outliers are represented by e@.

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Figure 2. From m left to right: P. striatulus oviposition puncture with pitch droplet (July), larval

galleries in phloem (October), foliage fade in October of subalpine fir trees attacked by P

striatulus earlier in the summer.

Observations and field sampling in 2016-2017 at the Watching Creek and Antler

Road sites revealed that larvae were active under the bark by late April or early May

from attack the previous summer. Larvae present near the phloem—sapwood interface

were beginning to construct chip cocoons at that time. Sampling found that the majority

of larvae within the bark layer did not score the sapwood. Slower development was noted

on the north aspect of trees, near the ground, and in trees located well within the stand,

where conditions were cooler and less sunlight could penetrate the canopy.

Overwintering adults were found in the duff layer at the base of trees throughout May

during the periodic field assessments. By mid-June, late-instar (large) larvae and pupae

were most predominant under the bark, and adults were observed on tree boles, cut

stumps, and in recent axe cuts on trees. From late June through mid-July, pupae and late-

instar larvae (Fig. 3) were predominant, and through July, teneral and mature adults were

found. Emergence peaked from late July into early September on trees that were attacked

the previous summer, although some larvae and a few pupae could still be found.

J. ENTOMOL. Soc. BRIT. COLUMBIA 117, DECEMBER 2020 9

Emergence holes were abundant on the southeast aspect (warmer side) of many trees

(Fig. 3). By October, all trees attacked by P. striatulus earlier in the year contained large

larvae (presumably third or fourth instar).

Figure 3. Life stages and signs of P. striatulus: eggs (top left); larva in chip cocoon (top

centre); pupa (top right); exit holes (bottom left); adult weevil in chip cocoon (bottom right)

prior to emergence; and mating adults.

Some trees attacked by P. striatulus in 2015 were subsequently attacked by WBBB

in 2016. Trees were already fading, but some available phloem remained for WBBB to

colonise. From late June through July, adult P. striatulus were observed mating and

Ovipositing on the boles of mature, live, outwardly healthy-looking subalpine firs at the

Watching Creek and Antler Road sites. The first adult emergence at the Watching Creek

site was observed on 11 July 2016. Therefore, weevils seen ovipositing could presumably

both be overwintered and newly emerged adults. By 20 July, many small larval galleries

were observed under the bark of newly attacked trees. On 5 July 2017, a large, live

healthy subalpine fir at the Antler Road site was observed undergoing mass attack, with

many adult weevils on the bole mating and ovipositing.

Temperature data were recorded throughout 2017 except for an 18-day period (25

May—12 June) when there was a malfunction, after which the temperature-recording

device was replaced. Mating, oviposition, eggs, and early instar larvae were first

observed when average daily minimum temperatures ranged from 5 °C to above 10 °C

and average daily maximum temperatures ranged from 15 °C to above 25 °C. Pupae

occurred when the average daily maximum temperature was from above 10 °C to the

mid-20 degrees Celsius (Fig. 1; Fig. 4). Little or no development was noted after average

daily minimum temperature was at or below 0 °C in late September to early October.

Based on trap catches at the Watching Creek site, two distinct flights by WBBB

occurred. one in late June and the other in mid-August to mid-September. The later flight

period was more prolonged. A few P. striatulus adults were caught in pheromone-baited

traps on 5 July and 13 July 2017. Weevils were caught at the end of WBBB’s first flight

period. Western balsam bark beetle were caught in traps once the average daily minimum

temperature rose above 0 °C and the average daily maximum temperature surpassed 10

°C, while P. striatulus were caught in traps once the average daily minimum temperature

surpassed 10 °C and the average daily maximum temperature was above 20 °C.

10 J. ENTOMOL. Soc. BRIT. COLUMBIA 117, DECEMBER 2020

Degree Celsius

~——- Min. temp.@2.5m ——Max.temp.@2.5m «Min. ground temp. ~~~ Max. ground temp.

Figure 4. Average daily minimum and maximum temperatures at Watching Creek in 2017

taken at ground level and 2.5 metres on the north side of a tree. The two red arrows indicate

when P. striatulus were caught in pheromone traps.

The three felled subalpine firs were just over 120 years old, and diameter at stump

height ranged from 21.0 cm to 43.0 cm. Tree 2, the largest tree, was attacked from the

base of the tree up to 21.0 m high and had the highest density of successful emergence,

with a maximum of over 100 exit holes per metre of? bark area at 6.5 metres height (Fig.

5). The average number of exit holes along the bole between 4.0 to 15.0 metres was 70

exit holes per metre of? bark area. The smallest felled tree (Tree 1), at 21.0 cm diameter

at stump height, had emergence from the stump end to 8.0 metres, ranging from 10 to 30

exit holes per metre of? bark area. In Tree 3 (31.0 cm stump diameter), most emergence

occurred from 2.0 to 10.0 metres height, ranging from 4 to 28 exit holes per metre of?

bark area (Fig. 5). Attack was found well into the crown area of all trees. The diameter of

the top section from each tree ranged from 9.0 cm to 18.5 cm.

A total of 1 202 larval head-capsule widths were measured, with widths ranging

from 0.4 mm to 2.2 mm (Table 2). The Hcap program (Logan et al. 1998) was used to

determine instar classification (Fig. 6). Hcap did not produce a clear separation of instars;

however, there appear to be four. The number of larvae within a range of head-capsule

widths was plotted, and four probable instar delineations were identified (Fig. 6). Based

on these delineations, the average head-capsule width (+ S.E.) and range were calculated

(Table 2). No clear demarcation occurred between second and third or third and fourth

instars, but the average size of fourth-instar head capsules was 1.6 + 0.15 mm (Table 2).

The length and width of 54 adult weevils and the width of 66 exit holes were

measured. The average length and width (+ S.E.) of P. striatulus was 6.7 + 0.1 mm and

2.6 + 0.05 mm, respectively. Exit holes were very circular and easily recognised, with an

average width (+ S.E.) of 3.3 + 0.1 mm (Fig. 3).

Pissodes striatulus was recorded in all 11 permanent sample plots previously

established to monitor mortality caused by WBBB and other damaging agents. Stems per

hectare of subalpine fir affected by the weevil ranged from 2 to 69, and tree mortality

ranged from <1% to 6%. The weevil was found both alone in trees and in combination

with WBBB. Incidence of the weevil varied by year and location, with two plots in the

ESSFxc and one plot in the ESSFwc having the highest recorded incidence of P

striatulus attack.

Field surveys. Fourteen geographic areas were surveyed for a total of 58 sites and

235 trees (Table 3) in the summer of 2017. Site elevation ranged from 1 257 m to 1 800

J. ENTOMOL. SOC. BRIT. COLUMBIA 117, DECEMBER 2020 11

m. One to eleven subalpine fir trees were assessed at each site, for an average of 4.1

+ 0.3 (+ S.E.) trees per site. Western balsam bark beetle attack was recorded at 69% of

the sites, and 60% of subalpine fir showed evidence of WBBB attack. Pissodes striatulus

attack was confirmed at 71% of sites, and of all subalpine fir assessed, 29% had some

level of weevil attack (Table 3). Nineteen per cent (45 trees) of the trees sampled were

colonised only by P. striatulus, and in the absence of any other potential mortality agent

(e.g., root disease), it was apparent that the weevil killed these trees.

120

ATree1 *Tree2 oTree3

100

Number exit holes per m?

Height on tree (m)

Figure 5. Number of P. striatulus exit holes (per square metre of bark) along the bole of three

mass-attacked subalpine fir trees.

DISCUSSION

This study reveals that P striatulus is a commonly found insect in subalpine

ecosystems and that it regularly attacks and occasionally kills subalpine fir in lower-

elevation, more climatically stressed stands in southern B.C. The weevil is likely

ubiquitous throughout all subalpine fir ecosystems in B.C.; it is recorded in this study’s

11 permanent sample plots, which are distributed throughout southern B.C., and in

northern B.C. (J. Robert, Ministry of Forests, Lands, Natural Resource Operations and

Rural Development, Omineca Region, B.C. personal communication), as well as from

Waterton north to near Grande Cache, Alberta (D. Langor, Canadian Forest Service,

Northern Forestry Centre, Edmonton, Alberta, Canada, personal communication).

Pissodes striatulus may act both as a primary attacker, killing live subalpine firs, and as a

secondary attacker, usually associated with WBBB. Both insects are known to colonise

trees exhibiting reduced vigour (Craighead 1950; Belyea 1952a; Bleiker et al. 2003)

caused by age (senescence), climate stress such as drought, or pre-existing stressors such

as defoliation or disease. This study’s observations show that P. striatulus and WBBB can

both initiate attack on apparently healthy, live subalpine firs, with one or the other

following in the secondary attacker role; both insects will attack freshly down trees or

stumps (L. Maclauchlan personal observation.; Stock 1991; McMillin et al. 2003, 2017).

The ability of the weevil to use downed trees and slash material and to switch roles

between secondary invader colonising highly stressed or dead trees and primary invader

attacking live, green trees demonstrates its capacity to adapt to changing and available

conditions. The live, green subalpine firs observed being attacked by P. striatulus did not

exhibit any outwards signs of stress or decline. Drought conditions may have predisposed

12 J. ENTOMOL. Soc. BRIT. COLUMBIA 117, DECEMBER 2020

trees to attack by one or both insects; however, no obvious drought mortality was

observed.

Density

<2.

Head capsule width (mm)

Figure 6. Head-capsule width distribution of P. striatulus. The line in the upper graph

represents possible instar distribution (graph generated using Hcap; Logan et al. 1998). The

lower graph shows frequency over a range of head-capsule widths and possible instar

separation.

Table 2. Delineation of P. striatulus larval instars by head-capsule width.

Rab sSNA a Sta Sa AS ae ea RS NE Ee A SCE A A i tit ha a a Bn SSI tl RS GN eS

Instar N Average width (mm) (+ S.E.) Size range (mm)

Nt 30 0.5+ 0.01 0.4-0.6

2 129 0.9+ 0.01 0.6-1.0

3 520 1340.01 1.1-1.4

4 523 1.6+0.01 bb e2u2

J. ENTOMOL. SOC. BRIT. COLUMBIA 117, DECEMBER 2020 13

Evidence that P. striatulus has killed or colonised trees may be difficult to detect.

Unlike WBBB, which leaves diagnostic gallery traces etched into the sapwood, P

striatulus rarely does so. The weevil often spends its entire development period within

the phloem—cork portion of the bark, where the traces of its galleries are quickly

obscured by those made by woodborers and other insects (L. Maclauchlan, personal

observation), that arrive after the weevil has initiated attack. Belyea (1952a, 1952b)

reported that P. striatulus was sometimes associated with the woodborer, Monochamus

spp. Dejean (Coleoptera: Cerambycidae) in balsam fir trees sampled in New Brunswick

and Quebec, Canada. and Minnesota, United States of America, after the severe eastern

spruce budworm outbreaks of the early 1900s. He suggested that P. striatulus could kill

weakened balsam fir trees that had survived severe budworm defoliation, up to four years

following the budworm outbreak. This supports the current study’s observations of P

striatulus contributing to the death of subalpine fir in ecosystems affected by disease or

defoliation or in years following drought events. Pissodes spp. Germar may live up to

four years (McMullen and Condrashoff 1973; Maclauchlan 1992; Langor and Williams

1998; Lewis et al. 2002), and this study records the presence of overwintered adults,

suggesting that P. striatulus populations could potentially build up when stressed trees

are abundant.

Trees attacked by P. striatulus alone have distinctive foliage-fade symptoms.

Treetops fade rapidly during the summer and begin to shed foliage, while the lower

branches of affected trees may turn red and lose foliage or they may retain some green

foliage. Crowns are generally characterised by browning and dropping of needles. By the

end of the summer, much of the foliage has dropped, and the attacked trees are a mix of

red and grey. This differs from the foliar-fade symptoms observed after WBBB attack,

which include the rapid, bright red colouration of foliage in the year following attack by

WBBB. Mortality associated with WBBB is caused by a beetle—fungus complex, WBBB

and Ophiostoma dryocoetidis (Kendrick and Molnar) de Hoog & Scheffer

(Ophiostomataceae) (Molnar 1965; Garbutt 1992; Bleiker et al. 2005), and the fungus

plays a vital role in the death of the host tree. There is no indication that P. striatulus has

a fungal associate, which could in part account for the different symptoms displayed by

the foliage of trees attacked only by the weevil.

Adult weevils are active from mid-June through to at least late August, showing a

long biological window for finding suitable hosts, mating, and oviposition. Adults were

found mating and ovipositing on trees before and throughout the emergence period of

new weevils from attacked trees. Therefore, both overwintered (older) weevils and new

adults could attack trees in mid-summer. The timing of attack can overlap with that of

WBBB, highlighting the possibility that either insect can be the first coloniser. Lewis ef

al. (2002) demonstrated that P. strobi (Peck), a significant pest of young Sitka spruce,

was capable of oviposition in the spring without needing to mate, if prior mating had

occurred the previous autumn and the females were fecund. This may explain, in part, the

long biological window. If long-lived adults are already fecund, synchronised emergence

is less necessary to find mates and colonise novel habitats. It may also explain why

pheromone-trap catches tend to be low in number for many species of Pissodes (Fontaine

and Foltz 1982; Phillips and Lanier 1986; Nevill and Alexander 1992; Miller and

Heppner 1999). Perhaps mating in this species occurs more frequently due to random

encounters rather than sexual attraction through pheromone release.

Pissodes striatulus will mass attack a tree from ground level to the upper crown of

trees. It prefers mature, large trees but, within a stand, can attack trees across a range of

sizes. Successful weevil emergence was noted from upper tree sections as small as 9.0

cm in diameter. Pissodes striatulus, like many Pissodes spp., displays a very plastic life

cycle, and development time can vary based on position of attack on the tree and annual

weather conditions. Development near the root collar was slower than it was on the main

bole and the warmer, east-facing aspects of attacked trees. Mating, oviposition, and larval

development progressed rapidly during the warmest summer period, when average daily

14 J. ENTOMOL. Soc. BRIT. COLUMBIA 117, DECEMBER 2020

minimum temperatures exceeded 0 °C and average daily maximum temperatures

ranged from above 10 °C to 30 °C.

The length of the 54 P. striatulus adults that were measured ranged from 4.9 mm to

8.3 mm, exactly comparable to the range of length of weevils described attacking

stressed balsam fir and spruce in eastern Canada (5.0-8.0 mm) (Swaine ef al. 1924). In

the east, oviposition is confined to the base or lower sections of the bole (Swaine et al.

1924; Belyea 1952a), whereas in the current study observations indicate that attack can

occur higher and along much of the length of the tree bole. The seasonal history and

timing of emergence vary in the literature (Swaine et al. 1924; Belyea 1952a), as they do

in this study, with adults emerging from June through August. Observations of this

weevil in eastern spruce—balsam fir forests suggest that the insect mainly breeds in

severely defoliated and nearly dead trees but are also able to attack trees that have

recovered from defoliation events (Belyea 1952b). Host selection parameters appear to

be similar on subalpine fir in B.C.; however, the stress level of trees and causal agents of

that stress are less obvious. |

Pissodes striatulus is a large Pissodes, with adults averaging 2.6 mm =< 6.7 mm

(width x length). Late-instar weevil larvae are larger than are bark beetle larvae

associated with the phloem of dying subalpine fir and so are easily distinguished. The

measurement of P. striatulus larval head-capsule widths did not clearly delineate instar

separation. There appeared to be four instars, averaging in size from 0.5 mm (first instar)

to 1.6 mm (fourth instar). Finding and collecting early instar larvae of this weevil was

difficult due to the cryptic nature of attack and oviposition by adults. Adult weevils are

difficult to see on tree bark, and the only visible sign of oviposition is a minute resin

droplet. Our frequent and rigourous field assessments allowed us to locate several trees

undergoing mass attack. However, dissecting out eggs and early instar larvae was

difficult. Therefore, early instars were underrepresented in our sampling. Some size

distinction occurred among later-instar head-capsule measurements, with a large range in

size. Much variation in the size of weevils and their brood occurred, due to parental

characteristics (large vs. small mothers), oviposition location on tree bole, age and size of

host tree, physiological condition of tree, and other biotic and abiotic influences; thus, the

size range of head capsules of successive instars overlaps. Most terminal-infesting

Pissodes have been reported as having four larval instars (Wallace and Sullivan 1985;

Park and Byun 1988; Langor and Williams 1998). Zhang er al. (2004) describe Pissodes

yunnanensis Langor et Zhang, a weevil that attacks boles of young Yunnan pine, Pinus

yunnanensis Franchet (Pinaceae), in southwestern China, as having four instars and the

head capsule width of the fourth instar averaging 6.1 mm. Finnegan (1958) reported that

P. approximatus (subsequently shown to be a synonym of P. nemorensis) (Godwin et al.

1982; Williams and Langor 2002) has four larval instars with head capsule widths

ranging in size from just over 0.3 mm to over 1.4 mm. Reports differ on the number of

larval instars of P. strobi; Harman (1970) describes five larval instars, with the average

head-capsule width ranging from 0.3 mm to 1.2 mm, whereas McIntosh ef al. (1996)

report four larval instars, with a similar range in average head-capsule width as Harman

(1970) from first through final instar.

In summary, P. striatulus is capable of mass attacking and killing large, mature

subalpine fir trees. Due to its abundance in lower-elevation subalpine fir stands, which

experience more frequent and severe drought events, we hypothesise that this abiotic

stress on the host tree attracts the weevil. Our findings show that adult weevils

overwinter, are likely long-lived, and are capable of oviposition over multiple years,

similar to other Pissodes species (McMullen and Condrashoff 1973; Furniss and Carolin

1977; Maclauchlan 1992). This would enable P. striatulus to take advantage of periodic

stressor events such as drought to build up populations rapidly. As moisture stress and

higher annual temperatures become prevalent in subalpine fir forests in B.C., the

presence and tree-killing habit of P. striatulus are likely to increase.

J. ENTOMOL. Soc. BRIT. COLUMBIA 117, DECEMBER 2020

Hyas Lake 50.7798° N, -119.9543° W 1,258 ll 32 22

Table 3. Survey results for P. striatulus showing geographic locations with general latitude and longitude (Lat, Long); elevation;

number of sites sampled per location; number of subs pine fir trees (BI) assessed; number of trees with WBBB attack; number of trees

where P. swiatibes was the sole ey a coloniser or co-occurred with WBBB B 2°) and total number of trees containing P. striatulus.

"Geographic No.s sites he Bl Ne BI oa: = ‘No. Bl I with weevil at attack

Location Lat/Long ampled sampled WBBB je attack ee attack Total

49. 8597° N, -120.1362° W 1,678

50.0326° N, -119.6752° W 1,360

Apedl Road 49.3865° N, -119.9458° W 1,800

Antler Road 50.8831° N, -120.4131° W 1,260

Crystal Mountain 49.8942°N, -119.8155° W 1,314

Sullivan 50.9843° N, -120.0796° W 1,357

as 50.0527° N, -119.6576° W 1,343

Badger Lake 51.0361° N, -120.1080° W 1,317

Peachland Mai 49.8090° N, -119.9985° W 1,469

Whiteman Creek 50.2476° N, -119.7161° W 1,378

Watching Creek 50.9084° N, -120.4392° W 1,274

T.F.L. 18 §1.7612° N, -120.0852° W 1,257 10 55 42

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16 J. ENTOMOL. SOc. BRIT. COLUMBIA 117, DECEMBER 2020

Pissodes striatulus is also found in trees under attack by WBBB, where it may

encounter less host resistance. Common tree defence mechanisms such as increased resin

flow and the production of defensive chemicals due to insect attack (Berryman and

Ashraf 1970; Alfaro et al. 2002) would be reduced if WBBB has already mass attacked

the tree, making it easier for the weevil to colonise unused portions of the bole. The

ability of P. striatulus to switch from secondary to primary invader depending upon

climate and the host conditions available makes it well suited to adapt to warmer and

more severe climate conditions. Subalpine fir is intolerant of high temperatures or

moisture deficits (Alexander 1987); therefore, as changes in climate continue, elevated

stress levels will continue in these outlying populations of subalpine fir, and potentially

in northern and high-elevation forests throughout B.C. Changing climatic conditions,

coupled with the fact that adult weevils are long-lived and may have multiple broods,

could allow the weevil to proliferate, colonise, and kill an increasing number of trees

throughout the range of subalpine fir, accelerating mortality and rates of succession. This

study emphasises the need for additional monitoring of and research into high-elevation

forests, their insect complexes, and how climate change can impact these fragile

relationships.

ACKNOWLEDGEMENTS

We thank K. Buxton and B. Zimonick for field assistance and Kamloops Fire Centre

Initial Attack Crew for felling trees. B. Zimonick also provided a photograph (Fig. 3.

mating adults) for inclusion in the manuscript. We also thank S. Neufeld, T. Griffin, and

K. Streichert for laboratory work. K. White and B. Bains provided thoughtful reviews of

the manuscript. We thank B. Bentz for her time analysing head-capsule data using the

Hcap model. Many thanks to the three reviewers for their thoughtful and meticulous

reviews of our paper. Their comments greatly improved our submission. The project was

supported in part by funding from the British Columbia Ministry of Forests, Lands and

Natural Resource Operations and Rural Development Forests Sciences Program.

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20 J. ENTOMOL. SOC. BRIT. COLUMBIA 117, DECEMBER 2020

Beetles in the city: ground beetles (Coleoptera: Carabidae)

in Coquitlam, British Columbia as indicators of human

disturbance

R. MCGREGOR! AND V. WAHL

ABSTRACT

Urban development may cause adverse effects on the ecological integrity of natural

areas in cities through habitat loss and fragmentation. Biological communities in

habitat fragments may be altered, which may, in turn, negatively impact ecosystem

services that contribute to the sustainability of urban areas. As such, methods are

required to assess anthropogenic impacts on urban habitats. Here, results are

presented of ground beetle (Coleoptera: Carabidae) monitoring in habitat fragments

in Coquitlam, British Columbia. Ground beetle diversity in Coquitlam is highest in

small, disturbed sites that include both native and introduced European species.

Several European carabid species are effective biological indicators of

anthropogenic disturbance in urban habitat fragments. Because of the relative ease

of collection and identification of carabids, monitoring of carabids by citizen

scientists can be used to assess human impacts on urban ecosystems.

Key words: ground beetles, Carabidae, biological indicators, urban ecology, insect

conservation

INTRODUCTION

The development and growth of cities depends on the natural environment and the

services it provides to human populations. The quality of human life in cities depends on

ecosystem services that are provisioning (e.g., food, water, fibre), regulating (e.g., air-

quality regulation, water purification, climate regulation, pollination) and cultural (e.g.,

spiritual, recreational and aesthetic values) (Ranganathan et al. 2008; Montserrat et al.

2010). An ecological approach to urban planning should optimize the function of urban

ecosystems in order to maintain ecosystem services and the resulting sustainability of

cities. This approach presents challenges, because urban development often has negative

consequences for ecosystem integrity. For example, urban development causes the

fragmentation and elimination of natural habitats, which can have adverse effects on

ecological sustainability (Fahrig 2003). In addition, urban ecological processes are, and

will continue to be, affected by global climate change (Bellard et al. 2012).

A rapidly increasing proportion of the world’s human population lives in cities. Only

10% of the human population lived in urban areas at the beginning of the 20th century,

but since 2008, over 50% of humans live in cities (Grimm et al. 2008). As this urbanizing

trend continues, an understanding of the ecology of urban areas is essential. Here, we

present the results of surveys of urban biodiversity in Coquitlam, a rapidly growing

community in the Metro Vancouver area of British Columbia, Canada. Our objectives

were (1) to determine the nature of ecological communities in urban habitat fragments,

and (2) to identify and develop biological indicator species to monitor ecosystem changes

caused by human activities and climate change. In particular, we studied the influence of

habitat fragmentation on communities of ground beetles (Coleoptera: Carabidae) in

Coquitlam parks.

| Institute of Urban Ecology, Douglas College, P.O. Box 2503, New Westminster, British Columbia V3L

5B2; mcgregorr@douglascollege.ca

J. ENTOMOL. SOC. BRIT. COLUMBIA 117, DECEMBER 2020 Zi

Ground beetles are members of the family Carabidae, which includes approximately

40,000 species worldwide. More than 3,000 of these species occur in North America

(Bland and Jaques 1978; Marshall 2018). Many species exhibit habitat specificity or

specificity at least to habitat attributes like moisture, temperature and shade (Rainio and

Niemela 2003; Koivula 2011). As such, a strikingly different beetle assemblage can be

collected at locations in close proximity if habitat characteristics vary.

Surveys of ground beetles have been widely used to assess the influence of human

activities on ecological integrity. Previous studies have investigated the influences of

forestry (Lemieux and Lindgren 2004; Latty et al. 2005; Pierce and Venier 2006; Work et

al. 2008; Bergeron et al. 2011), agriculture (Raworth et al. 1997; Prasad and Snyder

2006; Fusser et al. 2018) and urbanization (Magura et al. 2004; Hartley et al. 2007) on

ground beetle abundance and diversity. Ground beetles have been advocated as effective

biological indicators of human disturbance (Rainio and Niemela 2003; Pierce and Venier

2006). The beetles are easily collected, and many species can be identified by relatively

untrained observers. This allows the participation of municipal staff, students, and

members of the general public in monitoring programs.

We have sampled ground beetles in urban habitat fragments in Coquitlam since

2001. Beetle species captured include both native North American carabids and species

historically introduced from Europe (Spence and Spence 1988; Klimaszewski et al.

2012). We present results of ground beetle surveys in two different habitats that occur in

close proximity in Coquitlam — one, a severely human-impacted meadow area, and the

other, a fragment of native forest along a riparian corridor. These results indicate that

carabid communities vary widely between habitats that have differing levels of

anthropogenic influence. Our data suggest that carabid monitoring could be used to

assess the level of human disturbance in urban habitats. We also present comparisons of

beetle communities from several urban forest fragments in Coquitlam parks that vary

both in their level of human disturbance and in their geographic area. Our analysis of

these data asks whether particular carabid species can act as effective biological

indicators of anthropogenic disturbance. Our data show that monitoring for European

carabid species can be used to evaluate human impacts on urban forests. We also discuss

the use of biodiversity monitoring in public education — in particular, the potential

establishment of a citizen-science (Silvertown 2009) beetle-monitoring network in Metro

Vancouver.

MATERIALS AND METHODS

Ground beetle sampling. Pitfall traps were constructed using plastic beverage cups

(500 ml). A double-cup system was used where the outer cup had a drainage hole to

remove rainwater and the inner cup had a similar hole covered in plastic screening to

prevent escape of beetles. Traps were inserted into the soil at sampling locations such that

the brim of the trap was level with the surface of the ground. Traps were deployed at field

locations for one week, after which captured beetles were returned to the laboratory for

counting and identification. Beetles were identified using Lindroth (1961—1969b).

Beetle monitoring, 2001—2005. Surveys were conducted at two sites adjacent to the

David Lam campus of Douglas College in Coquitlam, in 2001, 2003, and 2005. One site

is dominated by meadow vegetation that has developed on the site of a former gravel

mine directly south of the campus (David Lam (DL) Meadow site, 49°17'12.69" N,

122°47'34.63" W). The plant community at this site is dominated by grasses, herbaceous

annuals (often introduced weeds), shrubs, and deciduous trees (predominantly red alder

(Alnus rubra Bong. (Betulaceae)) and black cottonwood (Populus balsamifera ssp.

trichocarpa) (Torr. & A. Gray ex Hook. (Salicaceae)). Native shrubs like hardhack

(Spiraea douglasii ssp. douglasii Hook. (Rosaceae)) occur, but introduced species

predominate (e.g., Himalayan blackberry (Rubus discolor Weihe & Nees (Rosaceae)) and

Scotch broom (Cytisus scoparius (L.) Link (Fabaceae))). The other site is a fragment of

De J. ENTOMOL. SOC. BRIT. COLUMBIA 117, DECEMBER 2020

native coniferous forest associated with Hoy Creek, a salmon-bearing stream (Hoy

Creek Forest site, 49°17'19.50" N, 122°47'38.39" W). Vegetation at this site is typical of

second-growth temperate rain forest in the Vancouver area, where the plant community is

dominated by western hemlock (Zsuga heterophylla (Raf.) Sarg. (Pinaceae)) and western

red-cedar (Thuja plicata Don ex D. Don (Cupressaceae)), with an understorey composed

of native plants, including several species of ferns (Polystichum munitum (Kaulf.) C.

Pres] (Dryopteridaceae), Pteridium aquilinum (L.) Kuhn (Dennstaedtiacae), Blechnum

spicant (L.) With. (Blechnaceae), and Dryopteris expansa (C. Presl) Fraser-Jenk.. &

Jermy (Dryopteridaceae)) and shrubs such as vine maple (Acer circinatum Pursh

(Sapindaceae)), red huckleberry (Vaccinium parvifolium Sm. (Ericaceae)), and salal

(Gaultheria shallon Pursh (Ericaceae)).

Paired traps were placed at the DL Meadow and Hoy Creek Forest sites for periods

of one week in September of 2001 (n=20), 2003 (n=25), and 2005 (n=26) during field

exercises of Douglas College Ecology classes (BIOL 2322 and 3305). Beetles were

identified and counted for each trap, and trap-catch data for all three years were pooled

for analysis (n=71 traps in each habitat). |

Beetle monitoring, 2008. Surveys were conducted in 2008 at the DL Meadow and

Hoy Creek Forest sites sampled in 2001, 2003, and 2005, and at eight additional forest

fragments in Coquitlam parks (Table 1). Park sites were either small (<10 hectares) or

large in area (>40 hectares) and were assigned to one of three levels of human

disturbance. High-disturbance sites experienced heavy human use (i.e., via trails adjacent

to busy residential areas and/or schools), medium-disturbance sites had less use by

humans—mostly via hiking on wilderness trails—and low-disturbance sites were

undeveloped forest fragments with limited human use. Five pitfall traps were installed at

20-metre intervals along a 100-metre transect arranged from the edge to the interior of

each site. Traps were installed at all 10 sites in July 2008 and were emptied once a week

at each site on a staggered rotation schedule until early September. Beetles were

identified and counted, and beetle-count data were pooled for all collection periods for

each trap, producing pooled counts at each site over the two-month period.

Data analysis. Abundances of the six most common carabids collected in 2001-—

2005 (Carabus granulatus Linné (CG), Carabus nemoralis Miller (CN), Calathus

fuscipes Goeze (CF), Pterostichus melanarius Illiger (PM), Scaphinotus marginatus

Fischer (SM), and Scaphinotus angusticollis Mannerheim (SA)) were compared between

the forest and meadow habitats using paired t-tests. Shannon diversity index was

calculated for each of the five trap locations at each of the 10 sites sampled in 2008 using

PC-ORD (MjM Software, Version 5). Shannon diversity index was compared among

sample sites, disturbance levels, and site areas (large vs. small) by single-factor analysis

of variance (ANOVA). Means were distinguished in ANOVA using Holm-Sidak tests

(p<0.05). Analysis of variance and t-tests were conducted using SigmaStat (Version

3.1.1);

Indicator species analysis (Dufréne and Legendre 1997) was conducted on the 2008

data using PC-ORD (Version 5) to identify carabid species consistently associated with

low, medium or high levels of human disturbance. This method calculates the observed

indicator value for particular species (IVobs) relative to groups within datasets (e.g., levels

of environmental factors of interest). Indicator value varies from no indication of a

particular group (0) to perfect indication (100). Statistical significance of IVobs is tested

using a Monte Carlo method, where sample units are randomly reassigned to groups and

indicator value ([Vran) is recalculated over a set of permutations (in tests presented here,

n=4,999 permutations). Significance value (p) is calculated as the proportion of

randomized trials, with indicator value equal to or exceeding the observed indicator value

(McCune and Grace 2002).

J. ENTOMOL. SOC. BRIT. COLUMBIA 117, DECEMBER 2020 23

Table 1. Ground beetle collection sites in Coquitlam, British Columbia, 2008

Site name Location Area Site Area Disturbane

(hectares) code code Code

DL Meadow 49°17'12.69" N 2.0 DL Small High

122°47'34.63" W

Hoy Creek 49°17'19.50" N 6.7 HO Small High

122°47'38.39" W

Scott Creek 49°16'44.95" N 3.0 S Small High

122°48'43.79" W

Harper 49°18'12.74" N ae | H Small Low

122°44'54.74" W

Walton forest 49°17'23.82" N 4.0 W Small Medium

122°48'19.06" W |

Eagle 49°18'51.31" N 42.2 E Large Low

Mountain 122°48'08.38" W

Ridge Park 49°18'36.12" N 59.5 R Large Low

| 22°47 27.0

Coquitlam 49°16'45.92" N 68.1 CR Large High

River 122°46'20.91" W

Mundy 1 49°15'00.99" N 178.3 M1 Large Medium

122°49'20.28" W

Mundy 2 49°15'47.43" N 17833 M2 Large Medium

122°49'28.94" W

RESULTS

Beetle sampling, 2001-2005. Trap catches from 2001—2005 were dominated by six

carabid species: four introduced European species (C. granulatus, C. nemoralis, C.

fuscipes, and P. melanarius) and two native North American species (S. marginatus and

S. angusticollis). Relative abundances of these six species in the forest and meadow

habitats are shown in Figure 1. Although mean abundance of C. granulatus did not vary

between habitats, mean abundances of the five other species were significantly different

between the Hoy Creek Forest and DL Meadow sites. Mean abundances of three

European species (C. nemoralis, C. fuscipes, and P. melanarius) were higher at the DL

Meadow site than at the Hoy Creek Forest site. Mean abundances of the native species, S.

marginatus and S. angusticollis, were higher at the Hoy Creek Forest site than at the DL

Meadow site.

Beetle sampling, 2008. Ground beetles from 17 species were captured in traps at the

10 sites sampled in 2008 (Table 2). The most common species collected was S.

angusticollis, accounting for 58% of all beetles captured. As in samples from 2001—2005,

beetles captured in Coquitlam in 2008 were a mixture of native species and species

introduced from Europe.

Shannon diversity index varied significantly among sites, with the highest diversity

recorded at the Hoy Creek Forest site (ANOVA: F=8.51, df=9, p<0.001; Figure 2). Traps

from the Hoy Creek Forest site yielded 14 of the 17 species collected in Coquitlam in

2008, including all of the European species and most of the native forest-dwelling

species. This site is small in area and has a high level of human disturbance due to its

location directly adjacent to a secondary school, apartment residences, and a college

campus.

24 J. ENTOMOL. SOC. BRIT. COLUMBIA 117, DECEMBER 2020

: mum Vieadow

a 3 Forest

Ss)

ote’

O 6

©.

od *

(ems

—

o. x

0 Gt VZLZZLZI

CG CN

Ground beetle species

Figure 1. Mean pitfall-trap catches of six ground beetle species in forest and meadow habitats

in the 2001—2005 beetle surveys. Trap catches are shown for Carabus granulatus (CG),

Carabus nemoralis (CN), Calathus fuscipes (CF), Pterostichus melanarius (PM), Scaphinotus

marginatus (SM), and Scaphinotus angusticollis (SA), where black bars are from the meadow

habitat and hatched bars are from the forest habitat. Paired bars marked with an asterix are

significantly different by t-tests (p<0.05).

Shannon diversity index did not vary among levels of human disturbance, but there

was a trend to higher diversity at more disturbed sites (ANOVA: F=2.84, df=9, p=0.07).

Shannon diversity index was significantly higher in small-area sites than in larger sites

(ANOVA: F=8.56, df=1, p=0.005). Higher diversity at small sites and at high-disturbance

sites occurred because beetle communities included both native forest—specialist and

European species.

Indicator species analysis revealed a significant association of five beetle species

with sites at the highest level of human disturbance (Table 3). All five species are

European in origin (C. granulatus, C. nemoralis, C. fuscipes, H. affinis, and P.

melanarius). No other beetle species were statistically significant indicators of any of the

three disturbance levels.

DISCUSSION

Ground beetle communities in Coquitlam, British Columbia, vary between human-

disturbed areas and fragments of undisturbed native forest. European species dominate

the ground beetle community at one highly disturbed meadow site, as they do in local

agricultural habitats (Raworth et al. 1997). Some forested sites have carabid communities

that include most of the common native and European species, whereas others have

communities mainly, or exclusively, composed of native forest species. Small, disturbed

forest sites have higher beetle diversity than large, undisturbed forest sites do because

they have been colonized by European species while retaining native species.

J. ENTOMOL. Soc. BRIT. COLUMBIA 117, DECEMBER 2020 25

Table 2. Pooled counts of ground beetles collected at all sites in 2008.

Species Total abundance

Scaphinotus angusticollis Mannerheim Lfoe

Calathus fuscipes Goeze* 176

Pterostichus algidus Leconte 176

Scaphinotus marginatus Fischer 149

Carabus granulatus Linné* 67

Omus dejeanii Reiche 49

Pterostichus herculaneus Mannerheim 49

Pterostichus melanarius Mlliger* 47

Carabus nemoralis Miiller* a7

Pterostichus pumilis Casey 3

Harpalus affinis Schrank*

Leistus ferruginosus Mannerheim

Cychrus tuberculatus Harris

Scaphinotus angulatus Harris

Loricera pilicornis Fabricus

Pterostichus lama Ménétries

Synuchus impunctatus Say |

TOTAL 1

*Species introduced from Europe.

m= MM WB A A Cf OO

Increased beetle diversity in areas invaded by adventive European species has been

frequently observed, arguably because the European species are, for the most part,

occupying previously unexploited synanthropic niches in urban environments (Spence

and Spence 1988; LaBonte 2011). In a study in southeast Australia, a higher diversity of

ants was measured in small habitat fragments compared to large habitat fragments in an

urbanizing environment due to the presence of more generalist ant species in small

fragments (Gibb and Hochuli 2002). Presumably, diet breadth of carabids should have a

similar effect on distributions. For example, mollusc-feeding forest specialists like S.

angusticollis are primarily restricted to fragments of temperate rain forest where their

preferred prey are available (Larochelle and Lariviére 2003; Marshall 2018), whereas

more generalist European species like P. melanarius can penetrate forested habitats in

addition to more urbanized environments (Niemela and Spence 1991).

Indicator species analysis has shown that the presence of European species in

Coquitlam carabid communities indicates high levels of human disturbance. This means

that urban forest fragments that have undergone more impact by human activity are more

susceptible to invasion by European generalist species. This may occur because human

contact has altered the forest habitats or because population pressure from European

species in adjacent anthropogenic habitats is greater. Regarding habitat alteration, it has

been shown that trampling by human traffic in urban forests has a strong influence on

carabid community composition (Kotze et a/. 2012). It is also important to note that our

data are derived entirely from pitfall-trap catches and that pitfall traps have been shown

to not accurately sample carabid communities. Pitfall catches are often biased towards

larger, more active, carnivorous species and deficient in small, phytophagous carabids

(Spence and Niemela 1994; Knapp ef a/. 2020). In our study, most carabids that were

26 a ENTOMOL. Soc. BRIT. COLUMBIA 117, DECEMBER 2020

captured were relatively large in size, and smaller taxa like Bembidion Latreille

species were not captured. Despite this, our survey method still has utility for assessing

human influence on urban forest fragments, because it is relatively inexpensive, it is easy

to implement, and it samples a representative proportion of carabid taxa.

Table 3. Indicator species analysis by disturbance level for 2008 ground beetle survey.

Observed indicator values (IVobs) for particular combinations of species and disturbance

levels are shown, as are mean randomized indicator values (IVran) from Monte Carlo

simulations (4,999 permutations). p is the proportion of IVran equal to or exceeding IVobs.

Species Disturbance IVobs Mean IVran p

Scaphinotus angusticollis Medium 38.0 a0.7 0.42

Calathus fuscipes * High 30.0 12.0 <0.01

Pterostichus algidus High 30.9 39.9 0.89

Scaphinotus marginatus High 31.4 30.6 0.40

Carabus granulatus * High 38.8 15.0 <0.01

Omus dejeanii Medium 20.0 9.4 0.07

Pterostichus herculaneus Medium ota 23.8 0.54

Pterostichus melanarius * High 29.4 13.1 0.02

Carabus nemoralis * High 35,6 13.7 <0.01

Pterostichus pumilis Medium 16.3 fais eas.

Harpalus affinis * High i es 12.9 0.01

Leistus ferruginosus Medium rt 12.0 0.46

Cychrus tuberculatus Low 10.0 10.2 0.53

Scaphinotus angulatus Medium 8.6 10.3 0.66

Loricera pilicornis High 10.0 ia 0.34

Pterostichus lama High 25 6.7 1.00

Synuchus impunctatus High 5.0 6.0 1.00

*Species introduced from Europe.

It has been argued that the practice of urban planning can be substantially improved

by incorporating ecological concepts and recognizing the dynamic nature of urban

ecosystems (Flores et al. 1998). Given this, carabid beetle monitoring can be used to

predict adverse effects of urban development and inform municipal planning decisions

when managing for ecosystem services and biodiversity (Angold et a/. 2006). We predict

that the use of European carabids as indicator species will detect ecological effects of

urban development with a degree of subtlety higher than simple site assessments of

urbanization. Anthropogenic effects will vary, depending on the nature of both the habitat

being developed and the development project itself, and biological monitoring may be

the only effective method to assess impacts. Beetle surveys can, therefore, be used to

assess the ecological health of natural areas like urban forest fragments and landscape

corridors (Li et a/. 2008), and post-development monitoring can assess negative impacts

and, in turn, inform future planning. In addition, carabid monitoring could be used to

assess the ecological value of synthetic habitats like green roofs (Maclvor and Lundholm

2010) and “near-natural forests” (Da and Song 2008).

J. ENTOMOL. SOc. BRIT. COLUMBIA 117, DECEMBER 2020 pes i

We will expand and continue our ground beetle monitoring program in Coquitlam

and other Metro Vancouver municipalities. Long-term monitoring of changes in beetle

communities will provide a record of environmental change under the dual influences of

urbanization and climate change. We expect that the composition of beetle communities

will change in the future, as species ranges expand to the north with warming climate.

Novel mechanisms for unexpectedly rapid range expansion have been identified for

several insect species in Britain under the influence of climate change (Thomas et al.

2001). We also expect that new introductions of beetle species will occur as a

consequence of international trade, as has historically been the case through ship ballast

material and plant nursery stock (Brown 1940; Spence and Spence 1988). For example,

we recently detected the European carabid, Nebria brevicollis (Fabricus) in Metro

Vancouver, a species previously unrecorded in western Canada (R.R. McGregor, H.

Goulet, and J.R. LaBonte, unpublished observations). Nebria brevicollis was accidentally

introduced into Oregon, where it was first recorded in 2007 and has presumably spread

north to British Columbia (Kavanaugh and LaBonte 2008; LaBonte 2011).

1.8

1.6

1.4

1.0

0.8

0.6

0.4

Mean Shannon diversity index

0.2

0.0

H W DL HO S E M1 M2 R CR

Sample site

Figure 2. Mean Shannon Diversity Index for 10 sample sites in the 2008 beetle survey. See

Table 1 for site codes and descriptions. Hatched bars are for small area sites (<10 hectares)

and black bars are for large area sites (>40 hectares). Histogram bars marked with the same

letter are not significantly different (Holm-Sidak test p<0.05).

28 J. ENTOMOL. SOC. BRIT. COLUMBIA 117, DECEMBER 2020

Involvement of citizen scientists in monitoring programs for introduced and invasive

species has been advocated to increase the capacity of resource managers to collect data

(Silvertown 2009; Crall et al. 2010). Because of the relative simplicity of ground beetle

trapping and identification, there is a strong potential for public involvement in beetle

monitoring programs.

Given that, we plan to establish a beetle-monitoring program for the general public

and in Metro Vancouver schools. The program would have the dual purpose of gathering

data on local ground beetle communities and educating the general public and local youth

about the ecology of urban areas. Such a program would be facilitated by the ease of

collection and identification of ground beetles and by our experience in offering

environmental education programs. Student-based biodiversity-monitoring programs are

part of a recent trend for increased public participation in citizen-science programs

(Silvertown 2009). Two examples of similar programs are the Jimbovane Outreach

Project in South Africa, where students monitor ant biodiversity (https://www0.sun.ac.za/

limbovane/) and the Leaf Pack Network in the United States, where members of the

public monitor stream ecology (https://leafpacknetwork.org/).

Our work on beetle diversity in Metro Vancouver forest fragments makes an

important contribution to the understanding of ecological communities in urbanizing

environments. We have developed a monitoring program that can detect the influence of

human activity on ecological health through the presence of alien species. Information

from our surveys can be used to predict the consequences of further urban development

and, in turn, to inform planning decisions. Participation by students and other citizen

scientists in this and other biodiversity monitoring programs educates the public about

the importance of natural areas for urban sustainability. Urban habitat fragments provide

important ecosystem services and maintain biological diversity in our cities. As such, it is

critically important to monitor these areas as development and climate change inevitably

proceed.

ACKNOWLEDGEMENTS

Many thanks to students in Douglas College BIOL 2322 and 3305 for trapping

beetles from 2001 to 2005. Thanks to Caresse Selk and Shannon Wagner, of the City of

Coquitlam, for assistance in locating collection sites and setting traps in 2008. We are

grateful for financial support provided for this project by Toronto Dominion Friends of

the Environment Fund and the Douglas College Scholarly Activity Fund.

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J. ENTOMOL. SOc. BRIT. COLUMBIA 117, DECEMBER 2020 at

Effects of trail pheromone purity, dose, and type of placement on

recruiting European fire ants, Myrmica rubra, to food baits

D. HOEFELE, J. M. CHALISSERY, R. GRIES, and G. GRIES!

ABSTRACT

Trail pheromones of ants guide nest mates to a food source. Applications of

synthetic trail pheromone could guide ants to poisoned food baits, which may

expedite the demise of nests and help control invasive ant species. The trail

pheromone of the invasive European fire ant (EFA), Myrmica rubra Linnaeus

(Hymenoptera: Formicidae), has previously been identified as 3-ethyl-2,5-

dimethylpyrazine. To facilitate its development as an operational EFA control tactic,

our objectives were to determine the effects of (1) pheromone purity (isometrically

pure or isomeric mixture), (2) pheromone dose [2, 20, 200, 2,000 ant equivalents

(AEs)], and (3) type of pheromone placement (pheromone encircling a food source

rather than leading towards it) on ant recruitment to baits. In laboratory binary

choice experiments, isomerically pure and impure trail pheromone prompted similar

recruitment responses of ants. The presence of pheromone, irrespective of dose,

enhanced the recruitment of ants to food baits, with the dose of 200 AEs eliciting the

strongest recruitment responses (2 AEs: 61% of foraging ants; 20 AEs: 57%; 200

AEs: 69%; 2000 AEs: 59%). Pheromone applied in a line leading towards the food

bait, but not in a circle surrounding a food bait, was effective in recruiting ants,

suggesting that 3-ethyl-2,5-dimethylpyrazine has a guiding but not an attractive

function to EFAs.

INTRODUCTION

Many ant species use a trail pheromone to coordinate foraging efforts (Billen and

Morgan 1998). When a foraging ant finds a food source, she returns to the nest

depositing trail pheromone along the route. Nest mates then use this pheromone trail to

find their way to the food source, reinforcing the trail in the process. Foragers of some

ant species may deposit more pheromone, and thus recruit more nest mates, when they

have found a high-quality food source (de Biseau et al. 1991; Czaczkes et al. 2015).

Essentially, nest mates make collective decisions about the food sources they want to

exploit. Trail pheromone—guided foraging is common in ants (Czacskes et al. 2015),

including the European fire ant (EFA), Myrmica rubra Linnaeus (Hymenoptera:

Formicidae) (Cammaerts-Tricot 1973).

In their native range in Europe, EFAs can live in single colonies with hundreds of

workers and several queens (Fokuhl et al. 2007) but often form multi-nest colonies and

super-colonies (large networks of interconnected nests) that eradicate rivalling ant

species (Huszar et al. 2014). European fire ants prey on small invertebrates and tend to

aphids, from which they collect honeydew. They also collect elaiosomes and disperse

seeds (Fokuhl et al. 2007). In their invaded range (the east and west coasts of North

America), the ants do not engage in nuptial flights (Groden and Drummond 2005) where

winged queens mate with males and then land elsewhere to start a new nest. Instead, new

nests occur by “budding” (queens and workers leaving the original nest and establishing

a new nest nearby) without nuptial flights. As a result, local population densities of EFAs

can become so high that they exterminate native ant species (Naumann and Higgins

2015). Because the species swarms and stings aggressively when disturbed, it can render

gardens, lawns and parks unusable (Garnas 2004; Saltman 2016).

! Department of Biological Sciences, Simon Fraser University, Burnaby, British Columbia V5A 1S6,

Canada; G. Gries, gries@sfu.ca

32 J. ENTOMOL. Soc. BRIT. COLUMBIA 117, DECEMBER 2020

European fire ants are efficient foragers, apparently locating resources faster than

other ant species (Holway 1999; Groden et al., unpubl. data, as cited in Garnas 2004). To

coordinate foraging efforts, the ants use a trail pheromone (3-ethyl-2,5-dimethylpyrazine)

that they release from the poison gland (Evershed et al. 1982). Individual ants follow

trails of synthetic pheromone (Evershed et al. 1981), but it is not yet known whether

synthetic pheromone can be exploited for EFA control.

Current EFA control methods are not very effective. Excavating nests and spraying

them with permethrins is labour intensive and may need to be repeated if one or more

queens have been missed in the process (Higgins 2017). Topical insecticide treatments of

nests are ineffective, because they kill only those ants that happen to be above ground at

the time of treatment. The concept of lethal food baits to control EFAs is appealing,

because they forage socially and through trophallaxis might effectively distribute any

lethal agent among nest mates (Brian and Abbott 1977). The efficacy of lethal food baits

likely hinges upon attractive food odorants and/or synthetic trail pheromone to guide

foraging ants to food baits.

The EFA trail pheromone component 3-ethyl-2,5-dimethylpyrazine is commercially

available (Acros Organics, Part of Thermo Fisher Scientific, New Jersey, United States)

as a mixture of two isomers: 2-ethyl-3,6-dimethylpyrazine (= 3-ethyl-2,5-

dimethylpyrazine) and 2-ethyl-3,5-dimethylpyrazine ( = 3-ethyl-2,6-dimethylpyrazine.

. This isomeric mixture is inexpensive and is therefore suitable for development as

an EFA trail pheromone lure. However, as non-natural pheromone isomers can interfere

with optimal behavioral responses of insects (Roelofs and Comeau 1971), it is important

to determine whether pure and isomeric 2-ethyl-3,5-dimethylpyrazine elicit comparable

trail-following behavior by the ants.

Responses of insects to natural or synthetic pheromone are typically dose-dependent.

Larger amounts of synthetic pheromone as trap lures often result in greater trap captures

of target insects (Collignon et al. 2019). However, there are exceptions. In many ant

species, the concentration of trail pheromone modulates the trail-following response of

nest mates (Evershed et al. 1982; Kohl et al. 2001; Morgan et al. 2006), with the highest

pheromone concentration not always eliciting the strongest response. Workers of the

Western carpenter ant, Camponotus modoc Wheeler (Hymenoptera: Formicidae), follow

a low-dose synthetic pheromone trail for a longer distance than they follow a high-dose

synthetic pheromone trail (Renyard et al. 2019). Moreover, leafcutter ants, Atta sexdens

sexdens Linnaeus (Hymenoptera: Formicidae), walk longer distances on low-dose

pheromone trails than on high-dose pheromone trails (Morgan et al. 2006). Dose-

dependent responses to synthetic trail pheromone have also been studied with EFAs

(Evershed et al. 1982) but only in the absence of a food source.

A lethal food source must be deployed together with synthetic trail pheromone to

achieve the demise of fire ant nests. The placement method of synthetic trail pheromone

likely determines its effectiveness for recruitment of ants to lethal food baits. Ants

deposit trail pheromone in a line to guide nest mates towards a food source (Cammaerts-

Tricot 1978), but for ant control it would be most efficient to apply pheromone directly to

the food bait rather than laying down a pheromone trail towards it. By simply adding trail

pheromone directly to lethal baits, bait consumption by Argentine ants, Linepithema

humile Mayr (Hymenoptera: Formicidae), increased and thus resulted in greater ant

mortality and lower ant activity in the field (Greenberg and Klotz 2000; Welzel and Choe

2016). However, the recruitment effect of this type of pheromone placement may depend

on both the volatility of the pheromone and the propensity of foraging ants to be attracted

to, rather than guided by, trail pheromones.

Our overall objective was to determine whether the synthetic trail pheromone of

EFAs (3-ethyl-2,5-dimethylpyrazine) can be deployed to increase recruitment of nest

mates to food baits. Our specific objectives were to determine the effects of (1)

pheromone purity (isometrically pure or isomeric mixture), (2) pheromone dose (2—2,000

J. ENTOMOL. Soc. BRIT. COLUMBIA 117, DECEMBER 2020 33

ant equivalents), and (3) type of pheromone placement (pheromone encircling a food

bait rather than leading towards it) on ant recruitment to baits.

MATERIALS AND METHODS

Colony collections. We collected EFA colonies in the spring and summer of 2016—

2018 from Inter River Park (North Vancouver, British Columbia), the Burnaby and

Region Allotment Garden (Burnaby, British Columbia), and the VanDusen Botanical

Garden (Vancouver, British Columbia). To locate nests, we walked in a transect while

disturbing the soil by shuffling our feet. Because the ants respond quickly when

disturbed, it is easy to locate a nest entrance. Wearing nitrile gloves to protect ourselves

from stings, we excavated about 30 cm? of soil surrounding a nest entrance, and placed it

in a large bucket (19 L, 38 cm tall x 30 cm diam.). We slowly sifted through this soil by

hand, collecting about 10 queens, 200-300 workers, and 50—100 larvae and pupae from

each nest. We transferred the ants to artificial nest housings (see below), producing and

maintaining a total of 41 colonies for laboratory bioassays.

Rearing of experimental ants. We kept ant colonies indoors in the Science

Research Annex (49° 16'33" N, 122° 54'55" W) of Simon Fraser University at a

temperature of 25 °C and a photoperiod of 12L:12D. The rearing protocol took into

account that colonies need both an enclosed nest housing (hereinafter referred to as the

“nest-box”) and a surrounding foraging arena to exhibit normal behavior (Drees and

Ellison 2002; Fig. 1A). Each nest-box consisted of a small plastic container (15 x 15 x 9

cm), two-thirds of which was filled with potting soil (Sunshine® Mix #4, Sungro,

Agawam, Massachusetts, United States). A 10-cm? hole in nest-box lids was covered

with plastic mesh (Lumite Saran fabric, 10 ml) to allow for ventilation and water misting

(see below). Each nest-box was placed inside a foraging arena, consisting of a plastic tote

(41 x 29 x 24 cm or 58 x 43 x 31 cm) fitted with a mesh-covered hole (10 < 10 cm) in

the lid to allow air flow. Ants entered and exited the nest-box through a 15-cm-long

Nalgene tubing (3.175 mm diam.; Nalgene 180 PVC non-toxic autoclavable Lab/FDA/

USB V1 grade; Thermo Scientific, Waltham, Massachusetts, United States). The 5-cm-

wide, upper-most rim of each foraging arena was coated with a 1:1 mixture of paraffin oil

[White, Anachemia, Lachine (Montreal), Quebec, Canada] and petroleum jelly (Vaseline)

to prevent ants from escaping.

Twice per week, we added one source of protein (canned tuna, cat food, dog food,

tofu, canned beans, canned chicken, dehydrated shrimp, anchovy paste, dead mealworms,

dead blow flies, dead crickets, sunflower seeds, pumpkin seeds, luncheon meat, or corned

beef), and one source of carbohydrates (canned oranges, apple slices, apple sauce, grapes,

candy, honey, sugar water, cranberry sauce, or raisins) to the foraging arena, thus

allowing ants to leave their nest-box and forage (Fig. 1A). We provided the large variety

of foods to minimize the possibility that ants “learned” to forage for a specific food type.

We provided water in a test tube fitted with a piece of cotton, which we replaced

whenever it became moldy or dry. Twice per week, we rehydrated nest-boxes by spraying

water through the mesh window.

General design of the binary choice biossary. We deprived colonies of food (but

not water) 5—7 days prior to bioassays, testing each colony (n = 14—20 depending on the

experiment; see Table 1) only once for each stimulus and separating bioassays by at least

24 h. We ran experimental replicates during the ants’ photoperiod in six circular arenas

(122 cm diam. <x 40 cm height) housed in a dedicated bioassay room at 22—23 °C (Fig.

1B). To initiate a bioassay, we temporarily closed the entrance tube to a nest-box with a

piece of cotton. We then removed this nest-box from the foraging arena (Fig. 1A) and

placed it in a circular arena, such that the nest entrance tube was perpendicular to two

strips of filter paper (each 30 x 3 cm) taped to the arena floor. To provide a food source

for foraging ants, we placed a mixture of macerated apples and mealworms (1:1 ratio; 2 g

total) on top of a circular piece of damp cotton (9 cm diam.) at the distal end of each

34 J. ENTOMOL. SOC. BRIT. COLUMBIA 117, DECEMBER 2020

strip. After the ants (in their transferred nest-box) had acclimatized in the bioassay

arena for 10 min., we treated—by random assignment—one of the two paper strips with

a 25-ul aliquot of synthetic trail pheromone dissolved in pentane and the other strip with

a pentane control. Immediately following the application of test stimuli, we opened the

nest-box entrance to initiate the bioassay. We terminated all experimental replicates after

2 h (when ant foraging activity peaked according to preliminary testing), at which time

we counted the number of ants present on food baits or cotton circles, using these data as

the response variable for statistical analyses. We collected all foraging ants by aspirator

or hand and returned them together with the nest-box to their original foraging arena. We

tested pheromone in ant equivalents (AEs), with a mean pheromone amount of 5.8 ng

occurring in a single worker ant (Cammaerts et al. 1981).

Figure 1. Illustrations of the various nest-box components and design for the study. A. The

set-up for maintaining ant colonies in the insectary annex consisted of a foraging arena (58 x

43 x 31 cm) (1), which housed the ants’ NEST-BOX (15 x 15 x 9 cm) (2) fitted with a

Nalgene tubing (3.1 mm diam, 15 cm long) (3) for nest entry and exit, and was provisioned

with sources of food and water presented in Petri dishes (4) and in form of a moist cotton plug

confining a water reservoir inside a test tube (5). B and C. The experimental designs for

testing the effect of synthetic trail pheromone on foraging decisions by ants. For each

replicate in Design B, the nest-box (2) was placed inside a large circular bioassay arena such

that the entry, and exit tubing (3) was perpendicular to two filter paper strips (30 <x 3 cm) (6),

each leading to a circular piece of damp cotton (7 cm diam.) with a food bait (7). For each

replicate in Design C, each deposit of food bait was surrounded by a circular filter paper strip

(15 cm diam., 2 cm wide) (8), one of which was treated with synthetic trail pheromone and

the other with a solvent control. |

J. ENTOMOL. SOc. BRIT. COLUMBIA 117, DECEMBER 2020 35

Table 1. List of research objectives (O) and stimuli tested in Experiments 1-3.

Experiment # Test stimuli (T) Replicates®

O;: Determine the effect of pheromone isomer

VEER RAE AAR AAA AAR A RAR AAA AA AAA RA AAA AAA A

] T1: pure pheromone’; T2: pheromone mixture? 14

(200 AEs* tested for both T; and T2)

- T1:2 AEs; T2: Solvent control 20

Ti: 20 AEs; T2: Solvent control 20

Ti: 200 AEs; T2: Solvent control 20

Ti: 2,000 AEs; T2: Solvent control 15

Os: Determine the effect of pheromone! placement

3 a0 Rien circle sea bait (200 AEs); T2: Solvent 20

control circle around bait

43-ethyl-2,5-dimethylpyrazine;

bmixture of 3-ethyl-2,5-dimethylpyrazine and 3-ethyl-2,6-dimethylpyrazine at a 1:1 ratio;

CAE = ant equivalent of trail pheromone (5 ng)

dtested as pheromone mixture (see b)

‘Replicates equal the number of ant colonies tested

SPECIFIC EXPERIMENTS

Experiment 1. Effect of 3-ethyl-2,5-dimethylpyrazine alone and in combination

with isomeric 3-ethyl-2,6-dimethylpyrazine on trail-following responses of ants. The

commercial source of the EFA trail pheromone component 3-ethyl-2,5-dimethylpyrazine

(Acros Organics, part of Thermo Fisher Scientific, New Jersey, United States) is an

isomeric mixture of 3-ethyl-2,5-dimethylpyrazine and 3-ethyl-2,6-dimethylpyrazine at a

1:1 ratio. To isolate the natural (EFA-produced) isomer from the commercial isomeric

mixture, we employed a high-performance liquid chromatograph (HPLC) (Waters

Corporation, Milford, Massachusets, United States) fitted with a Synergy Hydro Reverse

Phase C18 column (250 mm x 4.6 mm, 4 y; Phenomenex, Torrance, California, United

States) and operated by a HPLC System (600 Controller, 2487 Dual Absorbance

Detector, Delta 600 Pump). Eluting the isomeric mixture with a 0.75-mL7?! ™" flow of

acetonitrile separated the two isomers but without baseline resolution. By collecting only

the second half of the later eluting target isomer (3-ethyl-2,5-dimethylpyrazine) peak, we

could obtain material for bioassays with 83% to 93% purity.

To determine whether the non-natural isomer (3-ethyl-2,6-dimethylpyrazine) in the

isomeric mixture had any adverse effect on trail-following responses of EFAs, we used

the general two-choice bioassay design described above (Fig. 1B), and tested the isolated

synthetic trail pheromone component 3-ethyl-2,5-dimethylpyrazine alone [200 AEs

(1,000 ng per trail)] versus the isomeric mixture containing 3-ethyl-2,5-dimethylpyrazine

at the same amount.

Experiment 2. Effect of trail pheromone dose on trail-following responses of

ants. To determine the trail pheromone dose that elicits the strongest trail-following

responses by EFAs, we tested each of four doses of synthetic trail pheromone (10, 100,

1,000, 10,000 ng, equivalent to 2, 20, 200 and 2,000 AEs, respectively) dissolved in

pentane (25 wL) versus a pentane control (25 pL) (Table 1). We applied the pheromone

treatment stimulus and the solvent control stimulus in 30-cm-long streaks on two non-

36 J. ENTOMOL. SOC. BRIT. COLUMBIA 117, DECEMBER 2020

overlapping paper strips (each 30 =< 3 cm) secured in a straight line to the bioassay

arena floor (see general experimental design; Fig. 1B).

Experiment 3. Effect of trail pheromone encircling a food bait on ant

recruitment. To determine whether EFAs respond to trail pheromone encircling a food

source (Fig. 1C) rather than leading towards it (Fig. 1B), we modified the experimental

design of Experiments 1 and 2. We surrounded each of the two food baits with a circular

strip of filter paper (15 cm diam.; 2 cm wide; cut from a circular filter paper; Fig. 1C)

and treated one strip with synthetic trail pheromone (isomer mixture, 200 ng) and the

other with a solvent control. This revised binary choice experimental design took into

account that the effects of trail pheromone encircling a food source or leading towards it

could not be compared directly because the linear trail would start near the nest entrance

and thus immediately bias the recruitment response of ants.

STATISTICAL ANALYSIS

We analyzed the data of Experiment 1 (effect of pure and isomeric pheromone on

trail-following responses of ants) and of Experiment 3 (effect of trail pheromone

placement on trail-following responses of ants) in JMP (SAS Institute Inc., Cary, North

Carolina, United States) with a y?-test against a theoretical 50:50 distribution. We

analyzed the data of Experiment 2 (effect of trail pheromone dose (tested as ant

equivalents) on trail-following responses of ants] using a general linear mixed model

with a binomial distribution and a logit link function, using the GLIMMIX procedure in

SAS. Ant equivalents were a fixed effect, and nest origin was a random effect. The over-

dispersion in the model was accounted for by scaling the standard errors proportional to

the deviance. We ran a Tukey-Kramer multiple comparisons test for pairwise

comparisons between treatment groups.

RESULTS

Experiment 1: Effect of 3-ethyl-2,5-dimethylpyrazine alone and in combination

with isomeric 3-ethyl-2,6=dimethylpyrazine on trial following responses of ants.

There was no difference in the proportion of ants that were recruited to a food bait by the

trail pheromone 3-ethyl-2,5-dimethylpyrazine alone or in combination with isomeric 3-

ethyl-2,6-dimethylpyrazine (v7 = 0.016, n = 13, df = 12, p = 0.898; mean number of

responding ants + SE (mra + SE): 61.2 + 17.3; Fig. 2).

Pure

pheromone

lsomeric

pheromone

05 O04 05) 02 010) 24> Ge Os 04 OS 0.6

Proportion (+ 95% Cl) of ants responding

Figure 2. Mean proportion of European fire ants present in binary choice arena bioassays (see

Fig. 1B) on cotton pads with food bait in response to pure pheromone (3-ethyl-2,5-

dimethylpyrazine) or isomeric pheromone (3-ethyl-2,5-dimethylpyrazine and 3-ethyl-2,6-

dimethylpyrazine) applied to the paper strip leading to the food bait; y?-test, p > 0.05. The

number in the bar centre (1) represents the one replicate where the nest was not responding

(total number of replicates run: n = 14; mean number of responding ants + SE: 61.23 + 17.34).

J. ENTOMOL. Soc. BRIT. COLUMBIA 117, DECEMBER 2020 a7

Experiment 2: Effect of trail pheromone dose on trail-following responses of

ants (Exp. 2). The presence of trail pheromone did affect the recruitment response of

ants (F365 = 11.15, p < 0.0001). When trail pheromone was tested at 2 AEs, it recruited

61% of the foraging ants to the corresponding food bait (n = 20, t = 3.27, p = 0.002; mra

+ SE: 35.0 + 7.5; Fig. 3). Trail pheromone tested at 20 and 200 AEs recruited 57% and

69% of foraging ants, respectively (20 AEs: n = 19, t = 2.09, p = 0.04; mra + SE: 36.2 +

10.1; 200 AEs: n = 20, t = 5.99, p < 0.001; mra + SE: 63.4 + 12.4). At the high dose of

2,000 AEs, the effect decreased to 59% of foraging ants (n = 11, t= 2.27, p = 0.03; mra +

SE: 49.8 + 8.7). The recruitment effect of the 200-AE dose exceeded that of the other

pheromone doses tested (200 vs 2: t = 0.002, p = 0.01; 200 vs 20: t = -5.06, p < 0.0001;

200 vs 2,000: t = 4.34, p = 0.0003; Tukey-Kramer analyses).

Experiment 3: Effect of trail pheromone placement on ant recruitment to bait.

When trail pheromone was applied at the most effective dose (200 AEs) on a filter paper

strip encircling the food bait (Fig. 1C) instead of leading towards it (Fig. 1B), the

pheromone failed to recruit ants to the food bait (y7 = 0.003, n = 19, df = 18, p = 0.958;

mra + SE: 36.5 + 7.9; Fig. 4), suggesting that the ants did not sense the pheromone.

} \ Trail pheromone

lath ad eat EN eae ack (Ant equivalents [AEs])

*b (2000) *P<0.05

O8 04.03 62 05 © OY G2 G2 Ga as ace G7

Proportion (+ 95% Cl) of ants responding

Figure 3. Mean proportion of European fire ants present in binary choice arena bioassays

(Fig. 1B) on cotton pads with food bait in response to isomeric pheromone (3-ethyl-2,5-

dimethylpyrazine and 3-ethyl-2,6-dimethylpyrazine) applied at 2, 20, 200 or 2,000 ant

equivalents (AEs; 1 AE = 5 ng of 3-ethyl-2,5-dimethylpyrazine) to the paper strip leading to

the food bait, each pheromone dose tested versus a solvent control. An asterisk (*) indicates a

significant preference for the pheromone stimulus. General linear mixed model, p < 0.05; the

200 AE trail pheromone dose was more effective than all others in recruiting ants to the food

bait (Tukey-Kramer test adjusted for multiple comparisons, p < 0.05). The numbers in bar

centres represent the number of replicates where the nest was not responding (total number of

replicates run: n = 20 for each of 2, 20 and 200 AEs; n = 15 for 2,000 AEs; mean number of

responding ants + SE: 2 AEs: 34.95 + 7.45; 20 AEs: 36.21 + 10.18; 200 AEs: 63.40 + 12. 43;

2,000 AEs: 49.81 + 8.72).

Pheromone

No pheromone

05 "04 02.02 67. 0. 01.02 03, G4 05

Proportion (+ 95% Cl) of ants responding

Figure 4. Mean proportion of European fire ants present in binary choice arena bioassays on

food baits surrounded by a circular filter paper strip that was either treated or not treated

(control) with synthetic trail pheromone (1,000 ng of a synthetic mixture of 3-ethyl-2,5-

dimethylpyrazine and 3-ethyl-2,6-dimethylpyrazine) (Fig. 1C). The trail pheromone near the

food bait had no effect on recruitment responses of ants; (y?-test, p > 0.05. The number in the

bar centre (1) represents the one replicate where the nest was not responding (total number of

replicates run: n = 20; mean number of responding ants + SE: 36.47 + 7.91).

38 J. ENTOMOL. Soc. BRIT. COLUMBIA 117, DECEMBER 2020

DISCUSSION

Our data provide useful information for the development of synthetic trail

pheromone as a means for guiding foraging EFAs to lethal food baits. We predict that

trail pheromone-guided rapid location of food by foraging ants and transport to the nest,

coupled with food-sharing trophallaxis, will facilitate the demise of nests and help

control local EFA populations. Our study has addressed important questions about

pheromone purity, optimal dose, and placement that needed to be answered before

operational pheromone implementation.

Affordability of pheromone-based control tactics is a key factor in their development

and sustained use. Pheromone-based pest control tactics are typically species-specific,

causing fewer non-target effects. This, in turn, makes pheromone-based control tactics

more expensive than conventional insecticides that control a wide range of pest insects.

Low pheromone synthesis costs contribute to the affordability of pheromone-based

control tactics and can sometimes be achieved by producing a mixture of optical or

structural isomers rather than stereo-specifically pure pheromone. The commercially

available and relatively affordable source of the EFA trail pheromone contains not only

the trail pheromone 3-ethyl-2,5-dimethypyrazine but also 3-ethyl-2,6-dimethylpyrazine

as a non-pheromonal structural isomer. The presence of optical or structural isomers in

pheromone lures is known to sometimes interfere with the optimal effectiveness of the

pheromone. For example, the attractiveness of synthetic (+)-disparlure, the sex

pheromone of the gypsy moth, Lymantria dispar Linnaeus (Lepidoptera: Erebidae) (Bierl

et al. 1970), is reduced in the presence of its antipode (-)-disparlure in a racemic

pheromone lure (Miller et al. 1977). Similarly, tetradecenyl acetates with a double bond

near Cll added to the sex pheromone (Z)-11-tetradecenyl acetate of the red-banded leaf

roller, Argyrotaenia velutinana (Walker) (Lepidoptera: Tortricidae), greatly decreases

pheromonal attraction of male moths (Roelofs and Comeau 1971). In light of these

findings, it was important to determine whether a non-pheromonal isomer impurity (3-

ethyl-2,6-dimethylpyrazine) in the commercial source of the EFA trail pheromone had

adverse effect on trail-following responses of EFAs. As both pure and isomerically

impure synthetic trail pheromone prompted similar trail-following responses by EFAs

(Fig. 2), use of isomerically impure pheromone can now be considered for operational

development.

The amount of trail pheromone deposited by ants or applied experimentally affects

the trail-following response of nest mates, as shown in carpenter ants, Camponotus spp.

(Kohl et al. 2001, 2003; Renyard et al. 2019), the leaf-cutting ant Atta sexdens sexdens

(Morgan et al. 2006), and the EFA (Evershed et al. 1982). Also in our study with the

EFA, trail-following responses were pheromone dose-dependent. As little as 2 AEs of

trail pheromone (0.33 ng/cm) were sufficient to enhance recruitment of EFAs to food

baits (Fig. 3), but a dose of 200 AEs (33 ng/cm) was more effective, recruiting on

average 12% more foraging ants. The effect was still present, but decreased, with the

highest dose (2,000 AEs or 330 ng/cm) (Fig. 3). These data differ from a previous report

(Evershed et al. 1982) that a trail pheromone dose of only 0.0319 ng/cm triggered the

strongest trail-following responses. These differences are not surprising given that

pheromone behavior in ants is very context-dependent (Vander Meer and Alonso 1998).

Evershed et al. (1982) presented a circular trail to groups of 25 or 50 EFA workers in the

absence of a food bait, recording the ants’ responses for 15 min. We, in contrast, offered

an entire nest [at least 100 EFA workers per nest; 15—20 nests (see Table 1)] a choice

between two paper strips treated with either a solvent control or the EFA trail pheromone,

each strip leading to a food bait where we counted the number of recruited ants 2 h after

bioassay initiation. The decreased activity of the highest trail pheromone dose (2,000

AEs) may reflect the behavioural choices of ants to ignore seemingly overcrowded trails

that do not allow for efficient foraging (Dussutour et al. 2004). It also may be a result of

J. ENTOMOL. Soc. BRIT. COLUMBIA 117, DECEMBER 2020 39

sensory overload, an effect that has been used to disrupt foraging behavior in the

Argentine ant (Suckling et al. 2011; Sunamura et al. 2011).

Trail pheromones of ants may embody some (MOglich et al. 1974) or all (Vander

Meer et al. 1990) of the following functions: orientation induction (prompting trail

following by nest-mates), orientation (guiding foragers along trails), and short-range

attraction (attracting foragers to trails). These functions may be mediated by a single-

component pheromone or a multiple-component pheromone blend (Jackson et al. 1990).

To test whether the single-component trail pheromone of EFAs has not only a guiding

function but also an attractive function, we deployed either the trail pheromone or a

solvent control on a circular paper strip surrounding a food bait, each bait placed 30 cm

away from the nest entrance. The very similar numbers of ants recruited to these two

food baits (Fig. 4) provide evidence that the EFA trail pheromone does not function as an

attractant, despite its volatility. Keeping a low profile by using a trail pheromone without

attractive function may be advantageous in settings of high nest density, where ants could

otherwise readily eavesdrop on their neighbors’ pheromone trails and exploit them

(Chalissery et al. 2019)

Our experiments were not designed to explore whether the EFA trail pheromone has

an orientation-induction function (Vander Meer et al. 1990), prompting or initiating trail

following by nest mates. Based on Cammaerts-Tricot (1978), it seems that a component

of the Dufour’s gland may serve this function.

Future research will need to determine the efficacy of synthetic trail pheromone in

field settings and explore potential types of pheromone formulations (e.g., pheromone-

laden ropes) and modes of deployment, all coupled with lethal food baits.

ACKNOWLEDGEMENTS

We thank Jenny Cory, Robert Higgins, and three anonymous reviewers for

constructive comments; Robert Higgins for identification of ant species; Shelby Kwok,

Sebastian Damin, Stephanie Fan, Kris Cu, Jessica Chalissery, Nikalen Edwards, Ady

Zhang, Carlisle Shih, and Archit Amal for assistance with colony maintenance, field

collections, and experiments; Ian Bercovitz and Adam Blake for statistical consultations;

and Stephen Takacs for graphical illustrations and some statistical analyses. This research

was supported by a Vice President Research — Undergraduate Student Research Award to

JC, a Graduate Fellowship from Simon Fraser University, and a Thelma Finlayson

graduate fellowship to DH. The research was further supported by an NSERC-—Industrial

Research Chair to GG, with Scotts Canada Ltd. as the industrial sponsor. |

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42 J. ENTOMOL. SOc. BRIT. COLUMBIA 117, DECEMBER 2020

Geographic range and seasonal occurrence in British

Columbia of two exotic ambrosia beetles as determined by

semiochemical-based trapping

E. STOKKINK!, J.H. BORDEN’, L.M. HUMBLE? AND L.J.

CHONG‘

ABSTRACT

Two exotic ambrosia beetles, Trypodendron domesticum (L.) and Xyloterinus politus

(Say) (Coleoptera: Curculionidae: Scolytinae), were captured in 2004 in traps baited

with either ethanol alone or ethanol and the aggregation pheromone lineatin at

locations outside their known range. The range of 7: domesticum in British

Columbia is now known to extend along lower Fraser Valley as far north as Yale and

along the Highway 3 corridor as far east as Sunshine Valley. Xyloterinus politus was

not recovered east of Hope but was trapped as far north as North Bend in the Fraser

River Canyon. Neither species was found on the Sunshine Coast or on Vancouver

Island. Traps on the Simon Fraser University (SFU) campus captured 7: domesticum

as early as the week ending 17 February 2004. At both SFU and the University of

British Columbia’s Malcolm Knapp Research Forest (MKRF) in Maple Ridge, the

majority of 7: domesticum were captured well before peak flight of the native striped

ambrosia beetle, 7rypodendron lineatum (Olivier). The flight of X. politus occurred

much later, spanning the months of April and May. Catches of 4,716 T. domesticum

in three traps at SFU and 59 X. politus at the MKRF indicate successful

establishment of both species. In future, the presence of both species will demand

expert taxonomic identification as a prerequisite to implementation and

interpretation of pest management tactics to prevent ambrosia beetle damage on

conifer and hardwood logs and lumber.

Keywords: exotic ambrosia beetles, Trypodendron domesticum, Xyloterinus politus,

British Columbia

INTRODUCTION

Two exotic ambrosia beetles in the tribe Xyloterini (Coleoptera: Curculionidae:

Scolytinae), Trypodendron domesticum (L.) and Xyloterinus politus (Say), are established

in British Columbia (BC) (Humble 2001). The former species is European in origin and

has recently begun to infest living angiosperm trees, in addition to its normal dead and

dying angiosperm hosts (Gaubicher et al. 2003; Petercord 2006). It has adapted to attack

red alder, Alnus rubra Bong., in coastal BC (Humble 2001). Xyloterinus politus is native

to eastern North America (Wood 1982), infests both angiosperm and gymnosperm hosts,

and has adapted to attack western hemlock, 7suga heterophylla (Raf.) Sarg., in BC

(Henry 2004).

In surveying for the presence of exotic woodborers, Humble (2001) discovered that

T: domesticum was already established in the BC Lower Mainland when surveys began in

1995. It was first recorded in Washington State in 2008 (Haack and Rabaglia 2013). The

first X. politus in western North America were recovered from survey traps in Burnaby in

1 Woodstock Management Inc., 2065 Bluebell Terrace, Nanaimo, BC VOS 2P9

* Corresponding author: JHB aasuhing. 6552 Carnegie Street, Burnaby, BC V5B 1Y3, and Prateesor

Emeritus, Simon Fraser University, 8888 University Drive, Burnaby BC V5A 1S6;

jhbconsult@outlook.com

3 Pacific Forestry Centre, Natural Resources Canada, 506 Burnside Road West, Victoria, BC V8Z 1M5

4 ¢/o 2745 Yale St., Vancouver, BC VS5K 1C4

J. ENTOMOL. SOC. BRIT. COLUMBIA 117, DECEMBER 2020 43

1997 (Humble 2001) and from King County, Washington, in 1996 (Mudge et al.

2001). By 1999, both species were established in BC as far east as Ruby Creek, between

Agassiz and Hope (Humble 2001).

The objectives of this study were to determine if the two species’ ranges extended in

BC beyond those known by 1999 (Humble 2001) and to compare the species’ seasonal

activity trends in one urban forest and one rural forest site to that of the indigenous

striped ambrosia beetle, Trypodendron lineatum (Olivier).

MATERIALS AND METHODS

Geographic Distribution. Twenty-nine trapping locations were selected, mainly

along four corridors leading into the BC Interior: Highway 1 (northernmost location —

Lytton Lumber, south of Lytton: 50.2200° N, 121.5768° W), Highway 3 (easternmost

location — Highway 3A at Okanagan Falls: 49.3337° N, 119.5549° W), Highway 5

(northernmost location — rest stop at site of former toll plaza: 49.6307° N, 121.0158° W),

Highway 6 (one location — Riverside Forest Products millyard near Lumby: 50.2354° N,

119.0281° W), and Highway 99 (northernmost location — Seton Lake Road west of

Lillooet: 50.6688° N, 121.9799° W), as well as on the Sunshine Coast (northernmost

location — Saltery Bay: 49.7653° N, 124.3096° W) and Vancouver Island (southernmost

location — Jemico Enterprises log sort yard, Chemainus: 48.9079° N, 123.7450° W;

westernmost location — Sarita log sort yard on the Bamfield Road: 48.8828° N,

125.0351° W; northernmost location — Western Forest Products log sort yard near Port

McNeill: 50.5762° N, 127.1942° W). Trap sites were selected near roads or sites where

infested logs could have been transported or deposited and where angiosperm trees that

could serve as hosts for both species were growing.

At each location, two 12-funnel Lindgren traps were placed about 35 m apart. Lures

were obtained from Phero Tech Inc., Delta, BC. One trap was baited with a 40-cm-long

ethanol sleeve lure (release rate 30 mg/day at 20° C, as determined in the laboratory), and

the other was baited with an identical ethanol lure plus a flex lure releasing the

Trypodendron spp. aggregation pheromone lineatin (MacConnell et al. 1977; Schurig et

al. 1982; Klimetzek et al. 1981) at 0.02 mg/day at 20° C. Lures were positioned at the

midpoint of the funnel column to achieve a wide odour plume (Lindgren 1983).

Traps were set in place by 15 March in 2004 and 2005, with trap sites altered slightly

to maximize detection opportunity. Captured beetles were collected in late May and mid-

July, bagged, and frozen for later identification and counting using Bright (1976) and

Wood (1982) as definitive references. Sex of captured beetles was not determined. When

large numbers of 7Zrypodendron spp. were captured, volumetric estimates of numbers

were used (80 beetles/mL) instead of individual counting. Voucher specimens of 7:

domesticum and X. politus captured beyond the known geographic range (Humble 2001)

were deposited in the reference collection at the Pacific Forestry Centre, Natural

Resources Canada, Victoria, BC.

Seasonal Flight Activity. To determine the spring to mid-summer flight season for

each species, three pairs of 12-funnel Lindgren traps were set up on 9 February 2004 in

predominantly coniferous forest at the University of British Columbia’s Malcolm Knapp

Research Forest (MKRF), Maple Ridge, BC. Three more trap pairs were set up on the

campus of Simon Fraser University (SFU), Burnaby, BC, where red alder is the dominant

species. Both 7’ domesticum and X. politus had been previously collected at each

location. Traps were spaced 15 m apart within pairs, and pairs were spaced at least 35 m

apart. One trap in each pair was baited with ethanol, and the other, with ethanol plus

lineatin, as described above. Trap catches were collected weekly until 20 July and kept

frozen for later identification and counting. Trapping did not extend into August, when

the “sister” flight of re-emerged parent beetles would occur and the flight of emerged

brood beetles would begin. Catches in traps with both lures were pooled to compile

44 J. ENTOMOL. Soc. BRIT. COLUMBIA 117, DECEMBER 2020

seasonal trend data. Catches of native T. lineatum were also enumerated.

Temperature records were obtained from Environment Canada weather stations at both

locations.

Mean catches between traps baited with ethanol or ethanol plus lineatin were

compared by f-tests (a = 0.05) for 7; domesticum at SFU and_X. politus at the MKRF.

RESULTS

Geographic Distribution. Zrypodendron domesticum and X. politus represented

1.1% of the 91,339 ambrosia beetles captured, the majority of which (87,215) were T.

lineatum. Both X. politus and T. domesticum were found beyond their previously known

geographic range in BC (Table 1), the former species along the corridor of Highway 1

and the latter species along the corridors of Highways 1 and 3. Neither species was

caught north of the metropolitan area of Vancouver, in the Okanagan Valley, or on

Vancouver Island.

Table 1. Numbers of X. politus and T. domesticum captured in 2004 and 2005 in two locations

within and four locations beyond their known geographic ranges in BC. Catches within the

known geographic range are from the seasonal flight activity study.

Number captured

Geographic

range Location Year Lure ee L.

politus domesticum

Simon Fraser University, ethanol 3 v2

Burnaby, in forest south of

South Campus Road: 2004 ethanol

49.2252° N, 122.9123° W ati 0 3944

Within known lineatin

ee Malcolm Knapp Research ethanol 22 LO:

Forest, Maple Ridge, 1 km

from gate: 49.2569° N, 2004 ethanol

122.5564° W ea 37 18

lineatin

Highway 1, Hope River ethanol 16 0

General Store, 5 km south

of Yale: 49.5635° N, 2005 ethanol

121.4263° W + 6 2

lineatin

Highway 1, gas station ethanol 1 0

before bridge at east

boundary of Yale: 49.5627° 2004 ethanol

N, 121.4181° W ae 0 0

Beyond lineatin

known range —_ Across Fraser River from ethanol 3 0

Highway 1, Boston Bar

First Nation log sort yard, 2004 ethanol

North Bend: 49.8766° N, qe ] 0

121.4464° W lineatin

Highway 3, Sunshine ethanol 0 0

Valley, 22 km east of

Hope: 49.2720° N, 2005 ethanol

121.2274° W ‘i 0 15

lineatin

J. ENTOMOL. SOC. BRIT. COLUMBIA 117, DECEMBER 2020 45

Xyloterinus politus was found in three new locations and in both years of trapping,

while 7: domesticum was caught in two new locations in 2005. Most X. politus were

caught at two Yale locations, 20 and 25 km northeast of Ruby Creek — the previously

known easternmost distribution of the species. Four specimens were captured at North

Bend, a further 35.8 km north by road in the Fraser River Canyon from Yale. Twenty of

the 24 captured _X. politus were captured in ethanol-baited traps.

One 7: domesticum was captured at the southernmost Yale location, and 15 more

were captured at Sunshine Valley — 24 km east of Ruby Creek — and at an elevation of

914 m — 878 m higher than Ruby Creek. All were in lineatin-baited traps.

High capture numbers within the known geographic range indicated successful

establishment of both species (Table 1). Trypodendron domesticum predominated on the

SFU campus, where red alder is the most prevalent tree species, and _X. politus was the

most frequently captured species at the MKRF, where conifers predominate. At SFU,

significantly more 7: domesticum were captured in traps baited with ethanol and lineatin

than in ethanol-only baited traps (means + SE: 1,314.7 + 166.5 versus 257.3 + 60.7, N =

3, t= 6.0, df= 4, P= 0.004). At the MKRF, the difference in catches of X. politus in traps

baited with ethanol and lineatin or ethanol alone was not significant (means + SE: 12.3 +

4.4 versus 7.3 + 0.3, N =3, t= 1.0, df=4, P= 0.39).

Seasonal Flight Activity. At SFU, 7) domesticum was captured during the first

trapping period ending 17 February 2004 (Fig. 1), when the maximum temperature was

only 12° C. Fifty-five per cent of 4,716 7’ domesticum were caught before the first

T: lineatum specimens were collected three weeks later. The 90% catch level was reached

for T: domesticum by 27 April, whereas it took until 22 June for 90% of T: lineatum to be

captured. The three X. politus captured at SFU were caught on 20 April, 27 April, and 4

May.

3 250 Trypodendron lineatum

Trypodendron domesticum

Number Captured

Figure 1. Seasonal occurrence in 2004 on the campus of Simon Fraser University, Burnaby,

BC, of native Trypodendron lineatum and exotic Trypodendron domesticum, as determined by

catches in semiochemical-baited traps.

46 J. ENTOMOL. Soc. BRIT. COLUMBIA 117, DECEMBER 2020

The seasonal occurrence of 7: domesticum at the MKRF in 2004 was similar to that

at SFU, but far fewer beetles (28) were captured (Fig. 2). Although some 7. lineatum (36

beetles) were captured in the collections on 9 and 16 March, 68% of the catch of

T. domesticum had occurred before the first surge of 993 T: lineatum was collected on 23

March. Xyloterinus politus was first captured at the MKRF when the temperature reached

17° C, and flight extended from 13 April to 25 May (Fig. 2), coinciding with the flight

period at SFU (Fig. 1).

Trypodendron domesticum

Number Captured

Trypodendron

lineatum

17 24] 2 9 1623 30/6 13 2027/4 1118251 8 152229/6 1320

Number Captured

td ft

Oo wv

Figure 2. Seasonal occurrence in 2004 at the University of British Columbia’s Malcolm

Knapp Research Forest, Maple Ridge, BC, of native 7rypodendron lineatum and exotic

Trypodendron domesticum and Xyloterinus politus, as determined by catches in

semiochemical-baited traps.

DISCUSSION

This study establishes that the ranges of 7. domesticum and_X. politus in BC are more

extensive than previously known. Because past surveys (Humble 2001) did not extend

east or north of Ruby Creek, it is not possible to determine if the locations identified in

this study at North Bend and Sunshine Valley represent recent range expansions.

Xyloterinus politus, which can infest both angiosperm trees and conifers, likely has the

capacity to advance without human aid into the BC Interior along the Highway | (Fraser

J. ENTOMOL. SOc. BRIT. COLUMBIA 117, DECEMBER 2020 47

River Canyon) corridor. As well, sufficient angiosperm host trees probably exist to

enable the passage of T’ domesticum into the BC Interior by way of the Highway 1

corridor. 7rypodendron domesticum’s advancement into the interior along the Highway 3

corridor, which would demand passage through subalpine conifer forests at a minimum

elevation of 1,342 m, is less likely. Another possible route into the interior, which was

not sampled in this study, is by way of the Harrison Lake/Lillooet River Valley. The lack

of recovery of either species on Vancouver Island and north of the Vancouver area on the

mainland suggests that the Salish Sea and the large urban area, respectively, represent

significant barriers to range expansion in the absence of human aid. Despite the previous

collection of one X. politus on southern Vancouver Island by one of us (LMH,

unpublished), trapping at six Island locations failed to confirm establishment.

The high catch numbers of ZT. domesticum in traps baited with ethanol and lineatin

(Table 1) is consistent with the beetle’s use of lineatin as an aggregation pheromone in

Europe (Payne et al. 1983). Similarly, the very early winter—spring flight (Figs. 1, 2)

reflects the same type of early flight in Europe, where 7. domesticum flies at least one

month earlier than its congener, 7rypodendron signatum (F.) (Gaubischer et al. 2003).

The lack of any difference in X. politus catches in traps baited with ethanol or ethanol

and lineatin indicates that, unlike the majority of closely related Trypodendron species, it

does not respond to lineatin.

Both species represent diagnostic and potential pest problems. Until the arrival of

T: domesticum, T. lineatum and Gnathotrichus retusus (LeConte) (both mainly

coniferophagous species) were the only ambrosia beetles known to occasionally attack

and breed in red alder (Nijholt 1981; Kiihnholz ez al. 2000), a species with increasing

commercial uses (FPInnovations, no date). As the range of 7. domesticum expands in the

future, any infestation in red alder logs will thus require expert diagnosis in order to

implement a targeted pest management program. The response of 7: domesticum to traps

baited with ethanol and lineatin (Table 1) suggests that a mass-trapping program like that

being currently run for T. lineatum (Borden 1988; Lindgren and Fraser 1994) could be

implemented where 7: domesticum becomes a problem. However, the 9.5° C threshold

for flight (Petercord 2006) and very early peak flight in BC (Figs. 1, 2) demand that

mass-trapping be implemented as early as January. Because of cross attraction by native

T: lineatum, identification to the species level would be required in case later catches of

T: lineatum, which flies when temperatures reach 15.5° C (Borden 1988), create the

impression of an unrealistically large threat to the hardwood hosts of TZ. domesticum. In

turn, without expert separation of species captured, operational mass-trapping programs

directed at T. lineatum (Borden 1988; Lindgren and Fraser 1994) could be confounded by

early catches of I. domesticum, creating unjustified concern that valuable conifer logs

and lumber are at risk of attack much earlier than expected.

Because X. politus has adapted to attack western hemlock (Henry 2004), it may

adapt further to embrace other BC conifers as hosts. This may necessitate identification

to species level in the future, in order to implement species-specific pest management

programs. Because its seasonal flight period closely overlapped that of Gnathotrichus

sulcatus (LeConte) at the MKRF (data not shown), catches of X. politus responding to

ethanol could confound pheromone-based mass-trapping programs directed at

G. sulcatus.

REFERENCES

Borden, J.H. 1988. The striped ambrosia beetle. Jn Dynamics of forest insect populations. Edited by A.A.

Berryman. Plenum, New York. Pp. 579-596.

Bright, D.E.J. 1976. The insects and arachnids of Canada, Part 2: The bark beetles of Canada and Alaska,

(Coleoptera: Scolytidae). Canada Department of Agriculture, Ottawa. Publication No. 1576.

FPInnovations. (n.d.). Red Alder: Alnus rubra, Bong. Fact Sheet. Forintek Canada Corp. Vancouver, BC.

48 J. ENTOMOL. Soc. BRIT. COLUMBIA 117, DECEMBER 2020

Gaubicher, B., De Proft, M., and Gregoire, J.-C. 2003. Trypodendron domesticum and Trypodendron

signatum: two scolytid species involved in beech decline in Belgium. Jn Ecology, survey and

management of forest insects, Proceedings of the joint [UFRO meeting of Working Parties S7.03.06

“Integrated management of forest defoliating insects” and S7.03.07 “Population dynamics of forest

insects”, Krakéw, Poland, September 1-5, 2002. Edited by M.L. McManus and A.M. Liebhold.

United States Department of Agriculture, Forest Service, Northeastern Research Station. General

Technical Report NE-311. Pp. 134-135.

Haack, R.A. and Rabaglia, R.J. 2013. Exotic bark and ambrosia beetles in the USA: potential and current

invaders. Jn Potential invasive pests of agricultural crop species. Edited by J. Penna. CAB

International, Wallingford, U.K. Pp. 48—74.

Henry, L. 2004. Abundance and attack rate of exotic and native wood-boring insects in southwest British

Columbia. Bachelor of Science Honors thesis, University of Victoria, Victoria, BC.

Humble, L.M. 2001. Invasive bark and wood-boring beetles in British Columbia, Canada. Jn Protection

of world forests from insect pests: advances in research, papers presented at the XXI IUFRO World

Congress, Kuala Lumpur. Edited by RI. Alfaro, K. Day, S. Salom, K.S.S. Nair, H. Evans, A.

Liebhold, F. Lieutier, M. Wagner, K. Futai and K. Suzuki. IUFRO World Series Vol. 11, Vienna,

Austria. Pp. 69-77.

Klimetzek, D., Vité, J.P., and Konig, E. 1981. Uber das Verhalten mitteleurpadischer Trypodendron-Arten

gegen gegeniiber natiirlichen und synthetischen Lockstoffen. Allegmeine Forst- und Jagdzeitung,

152: 64-70.

Kiihnholz, S., Borden, J.H., and McIntosh, R.L. 2000. The ambrosia beetle, Gnathotrichus retusus

(Coleoptera: Scolytidae), breeding in red alder, Alnus rubra (Betulaceae). Journal of the

Entomological Society of British Columbia, 97: 103-104.

Lindgren, B.S. 1983. A multiple funnel trap for scolytid beetles (Coleoptera). The Canadian

Entomologist, 115: 299-302.

Lindgren, B.S. and Fraser, R.G. 1994. Control of ambrosia beetle damage by mass trapping at a dryland

sorting area in British Columbia. Forestry Chronicle, 70: 159-163.

MacConnell, J.G., Borden, J.H., Silverstein, R.M., and Stokkink, E. 1977. Isolation and tentative

identification of lineatin, a pheromone from the frass of Trypodendron lineatum (Coleoptera:

Scolytidae). Journal of Chemical Ecology, 3: 549-561.

Mudge, A.D., LaBonte, J.R., Johnson, K.J.R., and LaGasa. E.H. 2001. Exotic woodboring

Coleoptera (Micromalthidae, Scolytidae) and Hymenoptera (Xiphydriidae) new to Oregon and

Washington. Proceedings of the Entomological Society of Washington, 103: 1011-1019.

Nijholt, W.W. 1981. Ambrosia beetles in alder. Canadian Forestry Service Research Notes 1 (2): 12.

Payne, T.L., Klimetzek, D., Kohnle, U., and Mori, K. 1983. Electrophysiological and field responses of

Trypodendron spp. to enantiomers of lineatin. Journal of Applied Entomology, 95: 272-276.

Petercord, R. 2006. Der Flugverauf des Laubnutholzborenkafers Trypodendron domesticum L. 2002 bis

2004. Mitteilungen der Deutschen Gesellschaft fiir allgemeine und angewandte Entomologie, 15:

219-223.

Schurig, V., Weber, R., Klimetzek, D., Kohnle, U., and Mori, K. 1982. Enantiomeric composition of

‘Lineatin’ in three sympatric ambrosia beetles. Naturwissenschaften, 69: 602-603.

Wood, S.L. 1982. The bark and ambrosia beetles of North and Central America (Coleoptera: Scolytidae),

a taxonomic monograph. Great Basin Naturalist Memoirs No. 6, Bringham Young University,

Provo, Utah.

J. ENTOMOL. SOC. BRIT. COLUMBIA 117, DECEMBER 2020 49

Andrena (Melandrena) cyanura Cockerell (Hymenoptera:

Apoidea, Andrenidae), a valid North American species

C.S. SHEFFIELD!

ABSTRACT

Andrena (Melandrena) transnigra Viereck, 1904 (Hymenoptera: Apoidea,

Andrenidae), a species originally described from Seattle, Washington, is a large,

distinctive, and rather common solitary bee that is active in the spring and early

summer in western North America. Consideration of morphological variation

within females of this species across its range, particularly scopal hair colour,

with subsequent genetic analysis led to the discovery of two distinct DNA

barcodes attributed to this species; the 6.2% divergence between the sequences

was consistent with the distinctive morphology. As a result, A. cyanura

Cockerell, 1916 is here removed from synonymy with A. transnigra and

resurrected as a valid species. In addition, A. transnigra paysoni Cockerell, 1924

is also removed from synonymy with A. transnigra and is instead treated as a

new synonym of A. cyanura. The male of A. cyanura was previously described

as A. transnigra by Bouseman and LaBerge (1979), so a diagnosis is provided to

distinguish the two species; thus, the male of A. transnigra is treated for the first

time. Both sexes of A. cyanura are distinguished from A. transnigra and other

similar Melandrena Pérez, 1890. In addition to the morphological and genetic

differences between A. transnigra and A. cyanura, each also has a distinctive

geography in Canada, albeit overlapping in parts of British Columbia. Andrena

transnigra is seemingly restricted to the southern half of British Columbia,

whereas A. cyanura is more widespread, ranging from southern British

Columbia north to the Yukon and as far east as Saskatchewan. The limited

molecular data available for these species from the United States also supports

their status as distinct species, although re-examination of specimens in

collections will help to clarify their respective distributions in North America.

Keywords: Bees, morphology, DNA barcode, geography, resurrected taxon,

synonym

INTRODUCTION

Andrena Fabricius, 1775 (Hymenoptera: Apoidea, Andrenidae) is one of the largest

genera of bees, with 1 443 species recognised globally by Gusenleitner and Schwarz

(2002), although more than 100 additional species have been described and tallied since,

with 1 556 species currently known (Ascher and Pickering 2020). Dubitzky et al. (2010)

estimated that approximately 2 000 species likely exist globally, suggesting that about

25% of the species remain unknown or undescribed. In North America, there are

currently 471 species (Ascher and Pickering 2020; Sheffield 2020), at least 149 of which

occur in Canada (Sheffield et al. 2017). |

The Holarctic subgenus Melandrena Pérez, 1890 contains approximately 70 species

(Ascher and Pickering 2020), 24 of which occur in the Nearctic region (Bouseman and

LaBerge 1979). Melandrena are among the most common of the early season bees in

North America, with several species making important contributions to pollination of tree

fruit crops (e.g., Sheffield et al. 2003; Gardner and Ascher 2006; Park et al. 2016).

1 Royal Saskatchewan Museum, 2340 Albert Street, Regina, Saskatchewan S4P 2V7;

cory. Sheffield@gov.sk.ca

50 J. ENTOMOL. Soc. BRIT. COLUMBIA 117, DECEMBER 2020

Sheffield et al. (2017) published a summary of DNA barcoding efforts for the

Canadian bee fauna, noting several genera for which unique sequences (i.e., Barcode

Index Numbers, or BINs, as per Ratnasingham and Hebert 2013) have not yet been

attributed to a corresponding taxon name, with 19 of these BINs in the genus Andrena

alone. In addition, many bee species have been assigned multiple BINs (e.g., see

Sheffield et al. 2020) and some cases where multiple species share a BIN (e.g., Rehan

and Sheffield 2011; Gibbs 2018). These genetic differences usually correspond to

morphological or geographical differences that can have taxonomic and ecological

significance (e.g., Vickruck et al. 2011; Sheffield et al. 2016, 2020). |

In the Barcode of Life Data System (BOLD; Ratnasingham and Hebert 2007), two

BINs were attributed to Andrena transnigra Viereck, 1904. One of these

[BOLD:AAC1655] is found in southern British Columbia, ranging as far north as

Cheslatta Falls (53.763, —125.747), with specimens matching the typical form briefly

described by Viereck (in Viereck et al. 1904a). The second BIN [BOLD:AAC1656]

consists of specimens with females not true to the typical form of A. transnigra, although

they would certainly be identified to that species using the key of Bouseman and

LaBerge (1979). Specimens associated with the latter BIN are also more widespread in

western North America. The purpose here is to clarify the taxonomic status of

A. transnigra using morphological, molecular, and geographic information.

MATERIALS AND METHODS

Specimens in BOLD that were identified as A. transnigra and corresponded to BINs

AAC1655 and AAC1656, and other members of the subgenus Melandrena — A. regularis

Malloch, 1917 (AAC0276), A. nivalis Smith, 1853 (AAB5093), and A. vicina Smith,

1853 (AAC0275) were selected for analysis. In addition, two outgroups were selected —

A. (Taeniandrena) wilkella (Kirby, 1802) (AAA8959), and the colletid bee (Colletidae),

Colletes inaequalis Say, 1837 (AAE1758). All were reviewed for accuracy in taxonomic

identification by reviewing corresponding specimens held at the Royal Saskatchewan

Museum or photos on BOLD; no full DNA barcode sequences are yet available for A.

carlini Cockerell, 1901. To facilitate analysis within and between members of BINs

AAC1655 (11 specimens) and AAC1656 (23 specimens), members of the latter had their

names temporarily changed to Andrena sp. These vetted sequences (N=205, all > 600 bp)

were aligned using MUSCLE (Edgar 2004) within BOLD. Sequence divergence was

analysed with the Barcode Gap Analysis tool on BOLD, using the Kimura 2 Parameter

distance model and default parameters.

In addition to considering the morphology of the typical form of A. transnigra

described by Viereck (Viereck et al. 1904a, b), photographs of type material and

morphological descriptions of the taxa currently considered synonyms of A. transnigra —

A. cyanura Cockerell, 1916 and A. transnigra paysoni Cockerell, 1924 — were also

examined. Both taxa were placed into synonymy with A. transnigra by Bouseman and

LaBerge (1979).

To determine tentative distributional ranges of taxa considered here, data from

specimens identified as A. transnigra were downloaded from the Global Biodiversity

Information Facility (2020), with additional data added from specimens currently at or on

loan to the Royal Saskatchewan Museum, from BOLD, and from other online sources

where the identification could be confirmed (i.e., Naturalist, https://www.inaturalist.org).

Data were mapped using SimpleMappr (Shorthouse 2010). The dataset for the specimens

used in this study will be archived with Canadensys (http://community.canadensys.net/)

under resource title "Andrena cyanura, a valid North American species " and can be

accessed using the following: https://doi.org/10.5886/x6kwyje.

Photomicrography was undertaken with a Canon EOS 5D Mark II digital camera

with an MP-E 65 mm 1:2.8 1-5 macro lens (Canon Inc., Ota, Tokyo, Japan).

J. ENTOMOL. Soc. BRIT. COLUMBIA 117, DECEMBER 2020 51

Measurements were made with an ocular micrometer on a Nikon SMZ1000

stereomicroscope (Nikon Corporation, Minato City, Tokyo, Japan).

RESULTS

Analysis of the DNA barcode gap indicated that members of BIN AAC1655 (i.e., A.

transnigra s. str.) show 0.2% mean intra-specific variation (maximum 0.49%) and that

members of BIN AAC1656 show 0.58% mean intra-specific variation (maximum

1.86%), both being nearest neighbours to each other with 6.2% sequence divergence. In

contrast, the Melandrena pairs A. vicina (AAC0275) and A. nivalis (AAB5093) show

2.4% sequence divergence.

In addition to the genetic differences reported above, female members of AAC1656

differ from typical A. transnigra (per Viereck et al. 1904a) but match morphological

characters of type material and morphological characters provided in the original

descriptions of A. cyanura and A. transnigra paysoni, including the presence of pale

scopal hairs (Fig. la) and a bluish-tinged metasoma. Males corresponding to AAC 1656

match the description of A. transnigra provided by Bouseman and LaBerge (1979),

including the genitalia (Fig. 2) and hidden sterna (Figs. 3 — 4); however, males belonging

to AAC1655 (A. transnigra s. str.) show small but consistent differences from those in

AAC1656 (Figs. 2 — 4; and see below). Geographically, BIN AAC1656 has a distribution

that ranges further east (Saskatchewan) and north (Yukon) in Canada than that of the

more localised distribution of AAC1655 in southern British Columbia (Fig. 5). Based on

these morphological characters, geography, and molecular data, members of AAC1656

are being designated as A. cyanura, which is hereby removed from synonymy with A.

transnigra.

Figure 1. Scopal hair colour on the hind femur of females: A, Andrena cyanura Cockerell,

1916 — arrow points to mostly pale hairs, and B, A. transnigra Viereck — arrow points to dark

hairs.

SZ J. ENTOMOL. Soc. BRIT. COLUMBIA 117, DECEMBER 2020

Figure 2. Male genitalia, dorsal view (A, C) and lateral view (B, D) for A and B, Andrena

cyanura Cockerell, and C and D, A. transnigra Viereck. In A and C, the arrows point to the

shape of the apex of the basal lobe of the gonocoxite, and in B and D, the arrows point to the

concavity between the basal lobe and the gonocoxite.

Figure 3. Sternum 7 of males: A, Andrena cyanura Cockerell, and B, A. transnigra Viereck.

J. ENTOMOL. Soc. BRIT. COLUMBIA 117, DECEMBER 2020 53

Figure 4. Sternum 8, ventral view (A, C) and lateral view (B, D) for male A and B, Andrena

cyanura Cockerell, and male C and D, A. transnigra Viereck. Red boxes indicate the length to

width ratio of the base of the apical process in each species.

* transnigra s. |.

® cyanura

® transnigra s. str.

+ * cyanura types

oa * transnigra s. str. type

Figure 5. Distribution map. * = “Andrena transnigra Viereck s. 1.” (i.e., species recognised

here were not distinguished); red circles = A. cyanura Cockerell; red star = type localities for

A. cyanura and A. transnigra paysoni Cockerell; blue circles and star = A. transnigra s. str.

54 J. ENTOMOL. Soc. BRIT. COLUMBIA 117, DECEMBER 2020

Taxonomy

Andrena (Melandrena) cyanura Cockerell, 1916, new status

Andrena cyanura Cockerell 1916: 252 [92]

Lectotype & [designated here]. USA, Colorado, Troublesome, 8 June 1908, by S.A.

Rohwer, on Salix [California Academy of Sciences, no. 15325]. Photo of lectotype

examined.

Andrena transnigra paysoni Cockerell, 1924: 349 [2], new synonymy

Holotype °. USA, Wyoming, Chimney Rock, 17 May 1924, by C.L. Corkins

[United States National Museum (Smithsonian) no. 100707]. Photos of holotype

examined.

The BIN assigned to 4. cyanura is AAC1656. Although Bouseman and LaBerge

(1979) indicated a holotype for A. cyanura, Cockerell's original work (Cockerell 1916)

indicates two females sharing the same collection information, suggesting syntypes.

Because Bouseman and LaBerge (1979) did not provide sufficient information to

distinguish between the two syntype specimens and because a second specimen could not

be found during the present study, according to the International Commission on

Zoological Nomenclature (1999) Code Article 74.5 their work does not constitute a valid

lectotype designation, and following Code Recommendation 73F (“avoidance of

assumption of holotype’’) a lectotype is here designated. This corresponds to the material

at the California Academy of Sciences, catalogue no. 15325.

Diagnosis. The female of Andrena cyanura is most similar to other North American

Melandrena that have most of the pubescence on the legs, including the scopa, dark

brown to black, not pale, particularly Andrena nivalis, A. regularis, A. vicina, and

especially A. carlini and A. transnigra in Canada; it also resembles A. brevicornis

Bouseman and LaBerge, 1979 known from Texas. Andrena cyanura has a complete

flocculus (i.e., hairs long and curling so a complete semicircle is formed; Fig. 6b),

distinguishing it from A. nivalis and A. vicina, which each have an incomplete flocculus

(i.e., hairs shorter, not forming a semicircle; Fig. 6a). It differs from A. nivalis, A. vicina,

and A. regularis, which have pale pubescence on the pleura (Fig. 7a), by usually having

dark pubescence on the mesopleura (Fig. 7b—d) and medially on the scutum (Fig. 7c, d),

although some specimens from the Yukon have very few dark hairs medially on the

scutum; the thoracic pubescence of A. nivalis, A. vicina, A. regularis and A. carlini 1s

entirely pale (Figs. 7a—b), although A. carlini has dark hair on the pleura only (Fig. 7b).

Cockerell (1931) indicated that some of these Melandrena species (particularly A.

victima = A. vicina) have thinner pubescence on the dorsum of thorax, giving the

superficial appearance of a dark band. Andrena cyanura can be distinguished from A.

transnigra by the colour of the scopal hairs on the trochanter and femur, the extent of the

pale hair on the mesopleuron, and in some specimens, the colour of the metasoma: in A.

cyanura, most of the scopal hairs on the trochanter and especially the femur are pale (Fig.

la), but black (or at least dark) in A. transnigra (Fig. 1b), and in many specimens of A.

cyanura, a thin line of pale hair extends down the anterior edge of the mesopleuron

below the pronotal lobe (Fig. 7d), being largely dark on A. transnigra (Fig. 7c). In

addition, the dorsal surface of the metasoma of many A. cyanura specimens have a

distinctive bluish sheen under certain lights, being entirely black in A. transnigra and A.

brevicornis (or the latter with reddish reflections). Andrena cyanura can be further

differentiated from A. brevicornis by its larger size (body length 10-15 mm versus 9 mm

in the latter) and sparse punctures on the metasoma (interspaces > 2 puncture diameters)

but denser in A. brevicornis (interspaces = 1 puncture diameter).

J. ENTOMOL. Soc. BRIT. COLUMBIA 117, DECEMBER 2020 55

Figure 6. Flocculus on the hind trochanter of female A, Andrena vicina Smith (incomplete

flocculus) and B, A. transnigra Viereck (complete flocculus).

Males of A. cyanura are most similar to A. transnigra and should key out to that

species using Bouseman and LaBerge (1979), although they differ in the shape of sterna

7 and 8, and in the shape of genitalia. In both species, the median emargination of

sternum 7 is deep (Fig. 3), but in A. cyanura, each lobe is broadly and evenly rounded

(Fig. 3a; matching the illustrations for A. transnigra in Bouseman and LaBerge (1979)),

whereas in A. transnigra, each lobe is angulate, with the longer portion on the outer edge

(Fig. 3b); the hairs on the lobe of A. cyanura are just less than twice the width of the lobe

(Fig. 3a), whereas in A. transnigra the hairs are longer, exceeding twice the width (Fig

3b). Sternum 8 of A. cyanura has the base of the apical process short, U-shaped, about as

long as wide, with the apex short with lateral hairs surpassing it in length (Figs. 4a, b),

whereas it is 1.6 times longer than wide in A. transnigra, V-shaped, narrowing more

acutely in the apical third, with apex more elongate and surpassing the lateral hairs (Figs.

4c, d). The gonocoxite of A. cyanura has a shorter basal lobe with a truncate apex (Fig.

2a), resulting in a shallower concavity (i.e., concavity subequal to apical width of basal

lobe) between lobe and gonocoxite in lateral view (Fig. 2b), whereas that of A. transnigra

is more elongate and narrow, the apex evenly rounded (Fig. 2c) and with a deeper

concavity that is greater than the apical width of the basal lobe (Fig. 2d).

Distribution. Andrena cyanura ranges further east and north than A. transnigra

does, occurring in Colorado and Wyoming (type localities), north to western

Saskatchewan, Alberta, central British Columbia, and into the Yukon Territory (Fig. 5).

Andrena transnigra occurs only in southern British Columbia, in Canada, and in the

western (i.e., coastal) states of the United States.

Andrena (Melandrena) transnigra Viereck, 1904

Andrena transnigra Viereck, in Viereck et al., 1904: 191, 223 [2]

Holotype &. USA, Washington, Seattle, 17 April 1896, by T. Kincaid [Academy of

Natural Sciences, Philadelphia (Drexel University) no. 10300]

The BIN assigned to A. transnigra is AAC1655.

Diagnosis. See diagnosis of A. cyanura (above) for comparisons to other

Melandrena. Andrena transnigra can be distinguished from A. cyanura mainly by the

black scopal hairs on the trochanter and femur (Fig. 1b); in A. cyanura, most of the

56 J. ENTOMOL. Soc. BRIT. COLUMBIA 117, DECEMBER 2020

scopal hairs on the trochanter and especially the femur are pale (Fig. la). In A.

transnigra, most of the hair on the mesopleuron is dark (Fig. 7c), whereas in many

specimens of A. cyanura, a thin line of pale hair extends down the anterior edge of the

mesopleuron below the pronotal lobe (Fig. 7d). The dorsal surface of the metasoma of A.

transnigra is always black, whereas many A. cyanura specimens have a distinctive bluish

sheen under certain lights.

Males of A. transnigra and A. cyanura differ in the shape of sterna 7 and 8, and in

the shape of the genitalia; in both species, the median emargination of sternum 7 is deep

(Fig. 3), but in A. transnigra, each lobe is angulate, with the longer portion on the outer

edge (Fig. 3b), whereas in A. cyanura, each lobe is broadly and evenly rounded (Fig. 3a;

matching the illustrations for A. transnigra in Bouseman and LaBerge (1979)); on the

lobe in A. transnigra, the hairs are long, exceeding twice the width (Fig 3b), but they are

shorter in A. cyanura, just less than twice the width of the lobe (Fig. 3a). Sternum 8 of A.

transnigra has the base of the apical process 1.6 times longer than wide, V-shaped,

narrowing more acutely in the apical third with the apex more elongate and surpassing

the lateral hairs (Figs. 4c, d); in A. cyanura, sternum 8 has the base of the apical process

short, U-shaped, about as long as wide, with the apex short with lateral hairs surpassing it

in length (Figs. 4c, d). The gonocoxite of A. transnigra is more elongate and narrow, the

apex evenly rounded (Fig. 2c) and with a deeper concavity that is greater than the apical

width of the basal lobe (Fig. 2d), whereas that of A. cyanura has a shorter basal lobe with

a truncate apex (Fig. 2a) resulting in a shallower concavity (i.e., concavity subequal to

apical width of basal lobe) between lobe and gonocoxite in lateral view (Fig. 2b).

Figure 7. Lateral view of female Melandrena Pérez species: A, Andrena vicina Smith; B, A.

carlini Cockerell; C, A. transnigra Viereck — arrow points to the characteristic band of black

hairs across mesoscutum; and D, A. cyanura Cockerell — top arrow points to the characteristic

band of black hairs across mesoscutum, and bottom arrow points to the anterior area of pale

pubescence on the pleura extending below the pronotal lobe.

J. ENTOMOL. SOC. BRIT. COLUMBIA 117, DECEMBER 2020 57

DISCUSSION

Cockerell (1916) described A. cyanura from Colorado, United States of America,

indicating that the species had a shiny bluish tinge to the metasoma and that the

trochanter flocculus was white. Later, Cockerell (1924) described A. transnigra paysoni,

noting subtle differences from A. transnigra but also mentioning the band of light hair

extending down the mesopleura and the pale hairs on the femur (Fig. 7d; and see

holotype at: http://n2t.net/ark:/65665/m3cele184c-e84d-42 11-aacf-c484b3f674ab). In

both cases, Cockerell’s species were morphologically distinct, albeit subtly, from A.

transnigra from Seattle, which was briefly and inadequately described in the key of

Viereck (Viereck et al. 1904a). Lanham (1949) recognised A. cyanura as a distinct

species from A. transnigra, providing a key to females of the carlini species group.

The description of the female and male of A. transnigra s. 1. by Bouseman and

LaBerge (1979) is more consistent with A. cyanura than with the typical form — likely a

result of A. cyanura being more widespread (Fig. 5). Despite consistent morphological

differences in both the colour of integument and the scopal hair colour that are seemingly

linked in part to geographic distribution (Fig. 5), Bouseman and LaBerge (1979)

synonymised A. cyanura and A. transnigra paysoni under A. transnigra based on females

only because males were not described. As these morphological and geographic

differences correspond to consistent genetic differences in the mitochondrial cytochrome

c oxidase subunit I gene, A. cyanura is resurrected from A. transnigra, and A. transnigra

paysoni is considered a new synonym of the former. Although both A. cyanura and A.

transnigra occur in Canada, A. transnigra is restricted to southern British Columbia,

whereas A. cyanura, which also occurs in that province, is more widespread with its

range extending north into northern British Columbia and the Yukon Territories and east

to Saskatchewan (Fig. 5). Thus, A. cyanura is recorded as a new species from British

Columbia (see Sheffield and Heron 2018), and previous records of A. transnigra from

Alberta and Saskatchewan (Bouseman and LaBerge 1979; Sheffield et al. 2014)

represent A. cyanura.

Future phylogenetic work will further clarify the relationships among Melandrena,

although the phylogeny proposed by Bouseman and LaBerge (1979) for the North

American species suggests a close relationship between A. transnigra s. |. and A.

brevicornis, and also between A. regularis and A. carlini (i.e., the carlini group). The

carlini group of Lanham (1949) also included these species (excluding A. brevicornis,

which was not yet described) and A. cyanura, but it also included A. hurdi Lanham, 1949

and A. heterura Cockerell, 1930 (=A. hallii Dunning, 1898), which are now placed in

subgenus Jylandrena LaBerge, 1964 but with a suggested close affinity to Melandrena.

Andrena transnigra, aS recognised here, seems to be more closely related to A.

brevicornis than to A. cyanura, based on male genitalia and hidden sterna, although no

molecular data for A. brevicornis or A. carlini are yet available to further resolve this.

Although the limited molecular data available for A. cyanura and A. transnigra from the

United States also support their status as distinct species, re-examination of specimens in

collections will help to clarify their respective distributions in North America. For

example, Bouseman and LaBerge (1979) indicated that the scopal hairs on the hind femur

are usually pale, suggesting that A. cyanura is likely more common and widespread than

the typical form.

ACKNOWLEDGEMENTS

The author thanks Christopher Grinter and David Bettman (California Academy of

Sciences) for providing images and information on type materials during the challenges

of COVID-19, and also thanks two anonymous reviewers and Marla Schwarzfeld

58 J. ENTOMOL. SOC. BRIT. COLUMBIA 117, DECEMBER 2020

(Canadian National Collection of Insects, Arachnids and Nematodes, Ottawa,

Ontario) for helpful comments.

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Ascher J.S. and Pickering, J. 2020. Discover Life bee species guide and world checklist (Hymenoptera:

Apoidea: Anthophila) [online]. Available from http://www.discoverlife.org/mp/20q?

guide=Apoidea_species [accessed 27 January 2020].

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Hemisphere. Part IX. Subgenus Melandrena. Transactions of the American Entomological Society,

104: 275-389.

Cockerell, T.D.A. 1916. Some Rocky Mountain andrenid bees. The Canadian Entomologist, 48: 252—

254. https://doi.org/10.4039/Ent48252-7.

Cockerell, T.D.A. 1924. A bee collecting trip to Chimney Rock, Wyoming. Entomological News, 35:

347-351.

Cockerell, T.D.A. 1931. Rocky Mountain Bees. II. American Museum Novitates, 458: 1—20.

Dubitzky, A., Plant, J., and Schénitzer, K. 2010. Phylogeny of the bee genus Andrena Fabricius based on

morphology (Hymenoptera: Andrenidae). Mitteilungen der Miinchner Entomologischen

Gesellschaft, 100: 137-202.

Edgar, R. 2004. MUSCLE: multiple sequence alignment with high accuracy and high throughput.

Nucleic Acids Research, 32: 1792—1797. https://doi.org/10.1093/nar/gkh340.

Gardner, K.E. and Ascher, J.S. 2006. Notes on the native bee pollinators in New York apple orchards.

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https://doi.org/10.1664/0028-7199(2006)114[86: NOTNBP]2.0.CO;2.

Global Biodiversity Information Facility (GBIF). 2020. Andrena transnigra Viereck, 1904. Global

Biodiversity Information Facility Occurrence Download 29 June 2020. https://doi.org/10.15468/

dl.vdSd6g.

Gibbs, J. 2018. DNA barcoding a nightmare taxon: assessing barcode index numbers and barcode gaps

for sweat bees. Genome, 61: 21-31. https://dx.doi.org/10.1139/gen-2017-0096.

Gusenleitner, F. and Schwarz, M. 2002. Weltweite Checkliste der Bienengattung Andrena mit

Bemerkungen und Ergaénzungen zu Paliaarktischen Arten. Entomofauna, Suppl. 12: 1—-1280.

International Commission on Zoological Nomenclature. 1999. International Code of Zoological

Nomenclature. Fourth edition. International Trust for Zoological Nomenclature, London, United

Kingdom. xxix + 306 pp.

Lanham, U.N. 1949. Notes on the group of Andrena carlini Cockerell, with descriptions of a new species

from California (Hymenoptera: Apoidea). The Pan-Pacific Entomologist, 25: 33-35.

Park, M.G., Raguso, R.A., Losey, J.E., and Danforth, B.N. 2016. Per-visit pollinator performance and

regional importance of wild Bombus and Andrena (Melandrena) compared to the managed honey

bee in New York apple orchards. Apidologie, 47: 145-160. https://doi.org/10.1007/

s13592-015-0383-9.

Ratnasingham, S. and Hebert, P.D.N. 2007. BOLD: The Barcode of Life Data System (http://

www.barcodinglife.org). Molecular Ecology Notes, 7: 355-364. https://doi.org/10.1111/

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Ratnasingham, S. and Hebert, P.D.N. 2013. A DNA-based registry for all animal species: the barcode

index number (BIN) system. PLOS One, 8: e66213. https://doi.org/10.1371/journal.pone.0066213.

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Rehan, S.M. and Sheffield, C.S. 2011. Morphological and molecular delineation of a new species in the

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Sheffield, C.S. 2020. A new species of Andrena (Trachandrena) from the Southwestern United States

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Sheffield, C.S. and Heron, J.M. 2018. The bees of British Columbia (Hymenoptera: Apoidea,

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A., and Rowe, G. 2017. Contribution of DNA barcoding to the study of the bees (Hymenoptera:

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and designatable units of Bombus occidentalis Greene and B. terricola Kirby (Hymenoptera:

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60 J. ENTOMOL. Soc. BRIT. COLUMBIA 117, DECEMBER 2020

SCIENTIFIC NOTE

First record of the Palearctic seed bug Metopoplax

fuscinervis Stal (Hemiptera: Oxycarenidae) in North

America

C.G. RATZLAFF! AND G.G.E. SCUDDER?

Metopoplax Fieber, a small genus of seed bugs in the family Oxycarenidae

(Hemiptera), is naturally found throughout Europe, the northern coast of Africa, and

Central Asia. All three species of Metopoplax use plants of the Asteraceae family as their

host, from egg to adult (Péricart 1999). In Péricart (1999), Metopoplax ditomoides

(Costa) was recorded throughout the western Mediterranean region, extending north to

England and east to Bulgaria. Metopoplax fuscinervis Stal was recorded throughout the

entire Mediterranean region, extending as far north as central France and as far east as

Iran. Metopoplax origani (Kolenati) was recorded throughout the eastern Mediterranean

region and eastern Europe, extending eastwards into Central Asia as far as Kyrgyzstan.

Recently, MZ ditomoides was introduced to western North America, being found first in

Oregon in 1998 (Lattin and Wetherill 2002). Specimens were subsequently found in

California, Washington, and, in 2010, in British Columbia (Wheeler and Hoebeke 2012).

Expansion of the known range of M. fuscinervis has also occurred; it has recently been

recorded from Belgium (Baugnee et al. 2000), the Netherlands (Aukema et al. 2005), and

England (Harvey 2008).

On a warm, sunny day in August 2018, one male M. fuscinervis (Fig. 1) was

collected by the first author (C.G.R.) in Memorial South Park in Vancouver, British

Columbia, Canada. This specimen represents the first record of this species in North

America. It is deposited at the Spencer Entomological Collection in the Beaty

Biodiversity Museum at the University of British Columbia, Vancouver, British

Columbia, Canada.

Speciman data. @, CAN, BC, Vancouver, Memorial South Pk., 49.23117 °N,

123.08721 °W, 14.viii.2018 (C.G. Ratzlaff)

Metopoplax fuscinervis is easily separated from the two other species of Metopoplax,

in that the posterior half of its pronotum is lighter in colour (Fig. 1). In addition, it can be

distinguished from the other introduced species, M. ditomoides (Fig. 2), by its clypeus,

which is smaller and rounded or truncate instead of larger and spatulate. Péricart’s (1999)

key to the species of Metopoplax includes the variety M. origani var. cingulata and pairs

it with M. fuscinervis in the final couplet; however, the variety has since been

synonymized with M. fuscinervis (Dellapé and Henry 2020).

Metopoplax fuscinervis can be distinguished from other superficially similar species

present in the region using the key to Lygaeiodea in Henry (1997) and the key to genera

of Oxycarenidae in Henry ef al. (2015). In the latter, M. fuscinervis keys to couplet 8 but

does not agree with the characteristics for either Metopoplax, based on M. ditomoides and

differentiated previously, or Microplax, which is also based on a single species. The only

species of the genus Microplax recorded from North America is Microplax albofasciata

(Costa) (Hemiptera: Oxycarenidae), from California. Unlike M. fuscinervis, it has a

completely black pronotum (Henry et al. 2015).

1 Vancouver, British Columbia; chris.ratzlaff@gmail.com

2 c/o Spencer Entomological Collection, Beaty Biodiversity Museum, University of British Columbia,

2212 Main Mall, Vancouver, British Columbia. V6T 1Z4

J. ENTOMOL. SOc. BRIT. COLUMBIA 117, DECEMBER 2020 61

1mm

Figure 1. Male Metopoplax fuscinervis, a) dorsal view and b) lateral view, from Memorial

South Park, in Vancouver, British Columbia.

The specimen of M. fuscinervis was collected in a park in the middle of the city, in a

section of field only sparsely vegetated by short grass. This park is not close to any

obvious ports of entry, so how this specimen might have arrived at this location is

unknown. Little research has been done into the biology or dispersal of this species. The

recent recorded spread northwards in Europe suggests that M. fuscinervis is capable of

crossing larger distances, whether naturally or with human assistance. The closely related

M. ditomoides is thought to have been transported to new regions on plant material

(Deckert 2004). The same may be true for M. fuscinervis.

Vancouver is an apparent hotspot for introduced insects, with non-native species

consistently found there, many being new to British Columbia, Canada, or North

America. Hemiptera are no exception, with the aforementioned M. ditomoides,

Monosteira unicostata (Mulsant & Rey) (Hemiptera: Tingidae) (Scudder 2012),

Halyomorpha halys (Stal) (Hemiptera: Pentatomidae) (Abram et al. 2017), Cyphostethus

62 J. ENTOMOL. SOc. BRIT. COLUMBIA 117, DECEMBER 2020

tristriatus (Fabricius) (Hemiptera: Acanthosomatidae) (Ratzlaff and Scudder 2018),

and Aradus depressus (Fabricius) (Hemiptera: Aradidae) (Heiss and Scudder 2019) all

being reported in the city in the past eight years. In fact, several of the city’s most

commonly found insects, of any order, are introduced; these include Polistes dominula

(Christ) (Hymenoptera: Vespidae), Amphimallon majale (Razoumowsky) (Coleoptera:

Scarabaeidae), and the widespread Harmonia axyridis (Pallas) (Coleoptera:

Coccinellidae).

1mm

Figure 2. Female Metopoplax ditomoides, dorsal view, from Richmond, British Columbia.

Early detection and sampling are key to determining the extent of a species’

introduction and to estimating the introduction’s possible effects (Lodge et al. 2006).

Unfortunately, these new populations may already be well established and widespread

before they are detected, because collecting efforts in public spaces, such as city parks,

are usually minimal. In addition, many factors are taken into account in invasive species

management, and, for many reasons, land managers tend to view less noticeable species

that have few or no economic impacts as less of a management priority than invasive

species that have noticeable, economic impacts (Hanley and Roberts 2019). More

extensive sampling is needed to determine if M. fuscinervis has become established and,

if so, at what stage of introduction it is.

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J. ENTOMOL. SOC. BRIT. COLUMBIA 117, DECEMBER 2020 63

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some additional provincial and state records for Canada and the USA. Linzer Biologische Beitrage,

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64 J. ENTOMOL. SOC. BRIT. COLUMBIA 117, DECEMBER 2020

NATURAL HISTORY AND OBSERVATIONS

Plecoptera from the Crooked River, British Columbia

D.J. ERASMUS! and D.P.W. HUBER?’

The Crooked River is the southernmost Arctic watershed stream in British Columbia

(B.C.). It is the northwards-flowing outlet of Summit Lake, connecting a series of lakes

before it enters Williston Reservoir and the Peace River. With its location in north—

central B.C. and its south-to-north flow, the Crooked River is in the direct path of

petroleum pipelines originating from northeastern B.C. and Alberta to the Pacific coast.

One proposed pipeline, Coastal GasLink, is slated to cross the Crooked River (McCreary

and Turner 2018). Currently, no pipelines cross the Crooked River, and so it is important

to develop a species diversity baseline for future monitoring (Pauly 1995).

The EPT taxa richness index (Lenat 1988) — comprised of mayflies

(Ephemeroptera), stoneflies (Plecoptera), and caddisflies (Trichoptera) — is a monitoring

tool for the rapid assessment of water quality (Barbour et al. 1999). Most often, it is

compiled from larval data and generic-level identification, but it is also built from adult

specimens where species-level identification is possible. A baseline accounting of the

assemblage of EPTs in a river is thus useful for monitoring assemblage changes caused

by anthropogenic or natural changes (DeWalt et al. 1999). The order Plecoptera is the

most environmentally sensitive aquatic insect order to addition of organic pollutants

(Baumann 1979; Klemm ef al. 2002; Mandaville 2002) and monitoring shifts in species

presence may be important for assessing the effects of future oil, gas or bitumen spills.

We have previously published surveys of Trichoptera and Ephemeroptera for this

system (Erasmus et al. 2018; Huber et al. 2019), recovering at least 39 species of

caddisflies and 40 species of mayflies. We have reported a total of 11 new provincial

records for caddisflies and mayflies. This paper serves to complete the EPT checklist for

the Crooked River, because no previous work reports on stoneflies from this or nearby

systems.

Stonefly adults were sampled from May to August 2015 from eight locations (Table

1) using Malaise traps. Insects were captured into 95% ethanol, and the trap contents

were emptied every seven to 10 days. Collected specimens were transferred to new 95%

ethanol and stored at —20 °C until sorting.

Stoneflies were sorted to morphospecies and identified to the lowest possible taxon

using keys found in Stewart and Oswood (2006). Following sorting and morphological

identification, 95 adult specimens were sent to the Canadian Centre for DNA Barcoding

at the University of Guelph, Ontario. We obtained 92 useable sequences (>400 bp; <5

miscalls; no contamination detected). We identified species using the BOLD platform

with MUSCLE sequence alignments and a Kimura-2-parameter distance model. The data

set is available at dx.doi.org/10.5883/DS-CRPLE. Sequenced specimens, DNA, and

sequence data were vouchered at the Centre for Biodiversity Genomics and on the

Barcode of Life Database System at the University of Guelph.

A total of 3,421 adults were sampled, and based on our survey the Crooked River

supports at least 19 stonefly species belonging to 14 genera and seven families (Table 2).

Our initial morphology-based identification identified 13 species with a number of

specimens we classified only to family level, but DNA barcoding allowed the

identification of six more species. These data, combined with our previous efforts, result

1 Biochemistry and Molecular Biology, 3333 University Way, University of Northern British

Columbia, Prince George, British Columbia, Canada, V2N 4Z9; daniel.erasmus@unbc.ca

2 Faculty of Environment, University of Northern British Columbia, 3333 University Way, Prince

George, BC V2N 4Z9; huber@unbce.ca

J. ENTOMOL. SOC. BRIT. COLUMBIA 117, DECEMBER 2020 65

in a total count of 96 mayfly, caddisfly, and stonefly species for the Crooked River

(Erasmus et al. 2018; Huber ef al. 2019).

Ephemeroptera, Plecoptera, and Trichoptera species richness is a key indicator of

water quality (Kenney ef al. 2009), and the number of EPT species from Crooked River

compares favourably to other systems. Cordero et al. (2017) found 155 EPT species

across many rivers and lakes throughout northern Canada. Other studies have shown 70

EPT species from eight rivers of the Lower Illinois River basin (DeWalt et al. 1999).

Among these eight rivers, the highest EPT species count for a single river was 38 EPT

species. A survey for EPTs in the area around Churchill, Manitoba, revealed 112 EPT

species from several streams, ponds, and lakes (Zhou et al. 2009).

Our work provides a solid approximation for EPT species richness for the Crooked

River, B.C. It serves as a baseline for future studies such as environmental assessments. It

also provides a useful checklist for other systems in this rather under-surveyed region.

Table 1. Sampling Sites.

Site name Coordinates Description

CR2 54.484 °N, -122.721°W _ Tail out section at the end of a slow-moving

run and directly upstream from a bridge

crossing with a dirt road

CR2B 54.484 °N, —122.721°W Slow moving run with foliage up to the high-

water mark.

CR3 54.643 °N, -122.743 °W _ Riffle section, with foliage to the high-water

mark. 70 m from paved highway

CR4 54.388 °N, —122.633 °W _ Riffles and pools, with foliage to the high-

| water mark. 15 m from railway line and 40 m

from paved highway

CRS 54.478 °N, -122.719°W _ Riffles and runs with water supplied by

Livingston Springs.

CR6 54.328 °N, —122.669°W Outflow at Summit lake and directly

downstream from a bridge crossing with a dirt

road

CRIO0BR 54.446 °N, -122.653 °W _ Riffle, run, and pool section, and up- and

| downstream from a bridge crossing with a dirt

road. Foliage to the high-water mark.

CR108 54.458 °N, -122.722°W _ Riffle and pool sections, with foliage to the

high-water mark. 30 m from a dirt road.

J. ENTOMOL. Soc. BRIT. COLUMBIA 117, DECEMBER 2020

Table 2. Plecoptera collected from the Crooked River, British Columbia

Family?

Capniidae

Chloroperlidae

Leuctridae

Nemouridae

Perlidae

Perlodidae

Genus?

Capnia

Utacapnia

Alloperla

Haploperla

Sweltsa

Triznaka

Paraleuctra

Malenka

Podmosta

Zapada

Hesperoperla

Isoperla

Species?

coloradensis

Claassen, 1937

confusa

Claassen, 1936

columbiana

(Claassen,

1936)

severa

(Hagen, 1861)

brevis

(Banks, 1895)

coloradensis

(Banks, 1898)

signata

(Banks, 1895)

vershina

Gaufin and

Ricker, 1974

californica

(Claassen,

1923)

decepta

(Frison, 1942)

delicatula

(Claassen,

1923)

cinctipes

(Banks, 1897)

frigida

(Claassen,

1923)

pacifica

(Banks, 1900)

fulva

Claassen, 1937

sobria

(Hagen, 1874)

Specimen

Ip»

P5-CR108

and 1 other

P87-CR108

and 6 others

P1-CR108

P67-CR108

and 4 others

P48-CR3 and

3 others

P18-CR108

and 7 others

P71-CR2B

and 2 others

P37-CR108

and 2 others

P66-CR108

and 13 others

P55-CR4

P90-CR2B

and 14 others

P80-CR4 and

5 others

P54-CR4

P40-CR108

and 1 other

P59-CR4 and

10 others

P58-CR108

and 2 others

BINS

BOLD:A

AM4443

BOLD:A

AC4234

BOLD:A

AD2173

BOLD:A

ACSZ15

BOLD:A

AA6398

BOLD:A

AE SDO9

BOLD:A

AD7836

BOLD:A

DM0482

BOLD:A

AQ2350

BOLD:A

CG2351

BOLD:A

CS1790

BOLD:A

CB0726

BOLD:A

CC1688

BOLD: A

AK5931

BOLD:A

AC2488

BOLD:A

AL8080

Collectio

n site?

CR108

CR3

CR108,

CR2B,

CR4

CR108,

CR2B

CR108

CR108,

CR4

CR108,

CR2B,

CR3, CR4

CR108,

CR4

CR108,

CR4

CR108,

CR4,

CR2B

CR108,

CR4

J. ENTOMOL. Soc. BRIT. COLUMBIA 117, DECEMBER 2020 67

Table 2. Continued

Family? Genus? Species? Specimen BIN‘ Collection

ID» site?

Perlodidae Isoperla transmarina P22-CR3 and BOLD:A CR108,

(Newman, 3 others AA9661 CR3

1838)

Perlodidae Skwala americana P39-CR108 BOLD:A CR108

(Klapalek, AC6148

1912)

Pteronarcyidae Pteronarcys californica P41-CR108 BOLD:A CR108

Newport, 1848 AM2598

«Determined using morphological keys and BOLD database

> Specimen identification. Complete data set available at BOLD dataset CRPLE

¢ Barcode Index number

4 CR2: 54.484 °N, —122.721 °W; CR2B: 54.484 °N, —-122.721 °W; CR3: 54.643 °N,

—122.743 °W; CR4: 54.388 °N, —122.633 °W; CR5: 54.478 °N, —122.719 °W; CR6:

54.328 °N, —122.669 °W; CRIOOBR: 54.446 °N, —122.653 °W; CR108: 54.458 °N,

ie ice

ACKNOWLEDGEMENTS

We would like to thank Claire Shrimpton, Julie Shrimpton, and Dr. Mark Shrimpton

for their valuable assistance in the field.

REFERENCES

Barbour, M.T., Gerritsen, J., Snyder, B.D., and Stribling, J.B. 1999. Rapid bioassessment protocols for

use in streams and wadeable rivers: periphyton, benthic macroinvertebrates and fish (Vol. 339).

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Baumann, R.W. 1979. Nearctic stonefly genera as indicators of ecological parameters (Plecoptera:

Insecta). The Great Basin Naturalist, 39: 241-244.

Cordero, R.D., Sanchez-Ramirez, S., and Currie, D.C. 2017. DNA barcoding of aquatic insects reveals

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Erasmus, D.J., Yurkowski, E.A., and Huber, D.P.W. 2018. DNA barcode-based survey of Trichoptera in

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Great Lakes Entomologist, 32: 115-132.

Huber, D.P.W, Shrimpton, C.M., and Erasmus, D.J. 2019. Eight new provincial species records of

mayflies (Ephemeroptera) from one Arctic watershed river in British Columbia. Western North

American Naturalist, 79: 1-11.

Kenney, M.A., Sutton-Grier, A.E., Smith, R.F., and Gresens, S.E. 2009. Benthic macroinvertebrates as

indicators of water quality: The intersection of science and policy. Terrestrial Arthropod Reviews, 2:

99-128.

Klemm, D.J., Blocksom, K.A., Thoeny, W.T., Fulk, F.A., Herlihy, A.T., Kaufmann, P.R., and Cormier,

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conditions for streams in the Mid-Atlantic Highlands Region. Environmental Monitoring and

Assessment, 78: 169-212.

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macroinvertebrates. Journal of the North American Benthological Society, 7: 222-233.

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McCreary, T. and Turner, J. 2018. The contested scales of indigenous and settler jurisdiction: Unist’ot’en

struggles with Canadian pipeline governance. Studies in Political Economy, 99: 223-245.

Pauly, D. 1995. Anecdotes and the shifting baseline syndrome of fisheries. Trends in Ecology &

Evolution, 10: 430. Stewart, K.W. and Oswood, M.W. 2006. Stoneflies (Plecoptera) of Alaska and

Western Canada. Caddis Press, Columbus, Ohio.

Zhou, X., Adamowicz, S.J., Jacobus, L.M., DeWalt, R.E., and Hebert, P.D. 2009. Towards a

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Churchill, Manitoba, Canada. Frontiers in Zoology, 6: Article 30.

J. ENTOMOL. SOc. BRIT. COLUMBIA 117, DECEMBER 2020 69

NATURAL HISTORY AND OBSERVATIONS

New distribution records and range extensions of mosquitoes

(Diptera: Culicidae) in British Columbia and the Yukon

Territory

D.A.H. PEACH! AND L.M. POIRIER?

ABSTRACT We report the first records of (Diptera: Culicidae) Aedes euedes

Howard, Dyar, and Knab and Coquillettidia perturbans (Walker) from Canada’s

Yukon Territory, and the first record of Aedes decticus Howard, Dyar, and Knab

from British Columbia. We also report range extensions in northern British

Columbia for the western treehole mosquito, Aedes sierrensis (Ludlow), the

common house mosquito, Culex pipiens L., and the cool weather mosquito Culiseta

incidens (Thomson).

Key words: Aedes decticus, Aedes euedes, Coquillettidia perturbans, Culex pipiens,

Diptera: Culicidae, mosquito distribution

INTRODUCTION

Large-scale mosquito trapping for West Nile virus (WNV) surveillance has yielded

several additions to the mosquito fauna of British Columbia (BC) (Peach 2018b). This

effort logically focused on the south of the province, where most of the human

population is concentrated and where WNV is most likely to occur. In contrast, northern

BC and the Yukon Territory have experienced much lower survey effort, with bioblitzes

and individual collecting efforts largely responsible for advances in our knowledge of the

mosquito fauna in these areas.

Fifty mosquito species (Diptera: Culicidae) are currently known from BC (Peach

2018b), and 31 from the Yukon Territory (Belton and Belton 1990; Peach 2017, 2018a).

Here, we present new distribution records and range extensions from recent mosquito

collecting efforts, identified using keys to the mosquitoes of BC (Belton 1983), Canada

(Wood et al. 1979; Thielman and Hunter 2007), and North America (Darsie and Ward

2005), paired with supporting historical records where available and relevant. We

highlight the first records of Coquillettidia perturbans (Walker) and Aedes euedes

Howard, Dyar, and Knab from Canada’s Yukon Territory, and the first record of Ae.

decticus from BC. Although some of these records surely represent species that were

historically present and simply went undetected, others may represent more recent

extensions of northern range limits brought on by changing climate, anthropogenic

dispersal, or an increase of synanthropic habitat availability.

Aedes decticus Howard, Dyar, and Knab. Aedes decticus Howard, Dyar, and Knab

is an uncommon but widely distributed mosquito (Wood ef al. 1979). It lacks transverse

basal bands of pale scales on its abdominal tergites and has yellow and dark scales on its

vertex and postpronotum (Wood et al. 1979; Darsie and Ward 2005). Little is known of

its life history, but it has been reported in sphagnum bogs and other acidic woodland bogs

and pools (Mullen 1971). Aedes decticus has been reported from New England, regions

surrounding the Great Lakes, Labrador, northern Manitoba, and Alaska (Wood et al.

1979). Aedes decticus has been previously recorded from the Yukon Territory (Belton and

1 Department of Zoology, University of British Columbia, Vancouver, British Columbia V6T 124,

Canada; dan@danpeach.net

2 Department of Ecosystem Science and Management, The University of Northern British Columbia,

Prince George, British Columbia V2N 4Z9, Canada

70 J. ENTOMOL. SOC. BRIT. COLUMBIA 117, DECEMBER 2020

Belton 1990), but this has been overlooked in recent literature (Darsie and Ward

2005). Aedes decticus will take blood meals from humans (Smith 1952), but little is

known about what other sources of vertebrate blood it will take.

One adult female Ae. decticus was collected as it attempted to feed on DP at Wye

Lake, Yukon Territory, on 12 July 2019, and another as it attempted the same at a bog just

north of the town of Watson Lake, BC, on 13 July 2019 (Table 1). Two Ae. decticus

females were collected as pupae in muskeg beside a pull-off on the Alaska Highway in

BC, approximately 5 km southeast of Morley Lake, on 12 July 2019, and successfully

reared to adulthood. Another adult was collected attempting to feed on DP at Morley

Lake, BC, about 100 m south of the BC—Yukon border on 15 July 2019.

Aedes decticus is undoubtedly present on both sides of the border at the Morley Lake

location. While the extent of its range is unknown, the extensive spruce forest and boggy

conditions present in northern BC and the Yukon likely provide many localised areas of

suitable habitat for this species, and such conditions may extend from Alaska to

Labrador.

Table 1. Collection records of adult female or pupal mosquitoes from northern British

Columbia (BC) and the Yukon Territory (YT). Specimens are housed in 'the Beaty

Biodiversity Museum, University of British Columbia, Vancouver, BC, Canada, or *the Royal

British Columbia Museum, Victoria, BC, Canada.

No. of Date

Species Location Area Coordinates ere

pecimens

Aedes Town of Watson , 12 July

pane Lake, YT Wye Lake 60.067,-128.703 1 collected 019

Town of Watson Unnamed Hoovkd duly

Lake, YT bog 60.091, -128.738 1 collected 5019

Northern BC Morley Lake 59.999, -132.109 I collected! 13 1™Y

SE of 2collected 12 July

Northern BC Morley Aba 59.963, —132.046 asqupac! 019

Aedes Helmut’s , Ii July

ee ee Whitehorse, YT Pond 60.803, -135.097 1 collected 019

Seaforth eo 22 July

Southern YT teak 60.442, -133.551 1 collected 019

Southem YT —Liard River 60.020, ~128.604 1 collected! 1 Iuly

ieaieg ol HARESY Vac ts DAR Andie Sateeietelh, hides jk care Duly»,

ree Tanu Winida Cored; 53.025,-131.775 7 collected 1981

Lyell Island, on ) AWAY.

Gate Creek Haida Gwaii 52.667, —131.5 1 collected 1981

Louise Jul

Skedans Island, 52.964, -131.608 5 collected! 198 ,

Haida Gwaii

Hecate 3 Jul

Hotspring Island Strait, Haida 52.583,—131.433 3 collected! 1 Bed

Gwall

J. ENTOMOL. SOc. BRIT. COLUMBIA 117, DECEMBER 2020 i

Table 1. continued.

Species Location Area Coordinates S Ne. Date

pecimens

Hecate Strait, > 4July

Reef Island PE NEE 52.875, -131.52 1 collected 001

Hecate Strait, a

Reef Island aide Cian 52.875, —-131.52 3-collected 5001

Queen Charlotte i is , 25 July

City Haida Gwaii 53.256, -132.087 1 collected 019

Cranberry

Cuts Marsh/Starratt ee

‘ie Valemount, BC Wildlife 52.816, -119.268 1 collected! S:

pipiens Management 2004

Area

Cranberry

Marsh/Starratt 9 Sept

Valemount, BC Wildlife 52.819, -119.259 1 collected! “577”

Management

Area

Prince George, Danson z , 11 Aug.

BC Lagoon 53.831, -122.735 2 collected 2004

Prince George, Danson , 2/7 Aug.

BC Lagoon 53.831, —122.735 8 collected 2019

Culiseta Hecate Strait, Sips Se Dien ae ete 30 June

a taide Ross Island Paide (wait 52.163, -131.121 2 collected 1981

Lyell Island, aay

Gate Creek sidaiCwail 52.665, -131.466 3 collected 198]

Hecate Strait, 1 ae ky

Tanu Island ieide Celt 52.766, -—131.617 1 collected 198]

Hippa Island, , 24 July

Petrel Islet Haida Chesil 53.551,—-133.011 1 collected 019

pen Town Gv 12 July

Wye Lake Watson Lake, 60.065,-128.701 2 collected!

perturb YT 2019

ans

Albert Creek

: . Upper Liard, 3 observed, 13 July

bird banding YT 60.062, —128.917 o nattaataal 3019

station

Coal River , 1 14 July

Lodge Coal River, BC 59.657,—-126.956 2 collected 3019

pe J. ENTOMOL. Soc. BRIT. COLUMBIA 117, DECEMBER 2020

Aedes euedes Howard, Dyar, and Knab. Aedes euedes Howard, Dyar, and Knab is

a large, uncommon mosquito with bands of pale scales on the tarsi and scattered pale

scales on the proboscis, cerci, and posterior to the pale basal bands of the abdominal

tergites (Wood et al. 1979; Belton 1983; Darsie and Ward 2005). It lacks lower

mesepimeral setae and possesses reddish brown scales that form a broad mid-dorsal

stripe on the scutum (Wood ef al. 1979; Belton 1983; Darsie and Ward 2005). It is

thought to complete only one generation per year and to breed in permanent or semi-

permanent pools in a variety of habitats including woodland, marshes, and grassy areas

(Wood et al. 1979; Westwood et al. 1983).

Aedes euedes has been found in BC, Alberta, the Northwest Territories, and Alaska

(Wood et al. 1979; Belton 1983; Darsie and Ward 2005; Peach 2018b). Aedes euedes was

thought to likely be present but undetected in the Yukon Territory (Belton and Belton

1990). In summer 2019, adult females were collected trying to bite DP in an open area

near wastewater treatment lagoons outside Whitehorse, Yukon Territory, in the forest on

the margins of the Liard River, and along the margins of Seaforth Creek where it crosses

the Alaska Highway (Table 1). The latter area was a grassy riparian habitat that contained

numerous willows and small marshy areas with surrounding northern forest.

These collections represent the first confirmed records of Ae. euedes from the Yukon

Territory. This species is likely present at low numbers in suitable habitat throughout the

territory, as it has been found as far north as the coast of the Arctic Ocean in the

neighbouring Northwest Territories (Wood ef al. 1979).

Aedes sierrensis (Ludlow). The western treehole mosquito, Aedes sierrensis

(Ludlow), is a multivoltine species that breeds in water-filled holes in trees and can also

occasionally be found in water-filled artificial containers rich in plant debris and leaves.

Adults are black with striking bands of pale-yellow scales on the legs and patterns of the

same on the scutum.

Aedes sierrensis is an aggressive day-biter that takes blood from warm-blooded

animals, including humans (Peyton 1956), and is a vector of dog heartworm, Dirofilaria

immitis (Belton 1983). Aedes sierrensis is found over a broad portion of western North

America, from California to northern BC (Belton 1983; Darsie and Ward 2005),

wherever mature trees with suitable larval habitat are found. However, it was not

previously known to occur north of Terrace or on the islands of BC’s north coast.

Historical specimens in the Royal BC Museum indicate the presence of Ae. sierrensis in

the vicinity of Haida Gwaii; an additional specimen collected in August 2019 by C.

Stinson in Queen Charlotte City confirms the presence of this species in the area (Table

1). The range of Ae. sierrensis may also extend northwards into the Alexander

Archipelago.

Culex pipiens L. The northern house mosquito, Culex pipiens L., is a multivoltine

mosquito native to Eurasia that breeds in stagnant and polluted standing water, including

ditches, sewage lagoons, and water-filled containers rich in organic material (Wood ef al.

1979; Belton 1983). Larvae have an elongated siphon and are often found feeding at the

water’s surface. Adult Cx. pipiens are brown with bands of pale scales on the bases of

dark abdominal tergites. This species overwinters as nulliparous adult females that take

shelter in locations such as sheds, rodent burrows, tree bark, and rock piles until they

emerge to feed the following spring.

Culex pipiens blood-feed primarily from birds, although they will feed on humans as

well (Wood et al. 1979). These feeding habits make Cx. pipiens important vectors of

West Nile virus (Hamer et al. 2008). Culex pipiens have also been observed feeding on a

variety of nectar sources, including yarrow, Achillea millefolium, and common tansy,

Tanacetum vulgare (Asteraceae) (Peach and Gries 2020). Previously, Cx. pipiens had not

been reported north of southern BC (Belton 1983; Darsie and Ward 2005); however, in

2004, two specimens were captured in a CDC (Centres for Disease Control) blacklight

trap in Valemount, BC, by T. Brown, and two more at a sewage lagoon in Prince George,

J. ENTOMOL. SOC. BRIT. COLUMBIA 117, DECEMBER 2020 73

BC. Follow-up trapping in 2019 by LP at the Prince George site resulted in the

capture of an additional eight adult female Cx. pipiens (Table 1). Suspected Cx. pipiens

were observed by DP in Kitimat during September 2019, but they evaded capture.

It is likely that cold northern winters restrict the northern limits of Cx. pipiens

distribution. However, because the species will overwinter in heated artificial structures,

it is possible that it can be found in human settlements with suitable breeding conditions

much farther north than would be expected, because of warm outbuildings and human

transportation of equipment or other materials that may contain adults or eggs.

Culiseta incidens (Thomson). The cool weather mosquito, Culiseta incidens

(Thomson), is BC’s most common and widespread mosquito species (Belton 1983).

Culiseta incidens is a large mosquito with aggregations of dark scales on the wings, very

narrow bands of pale scales on the tarsi, and middorsocentral spots of pale scales and

bands of dark and pale scales on the scutum (Belton 1983). This mosquito breeds in

artificial containers, ditches, polluted water, storm drains, woodland pools, and coastal

and riverine rock pools. Females overwinter in warm, dry locations, such as in sheds,

under tree bark, in rock piles, or in rodent burrows.

Culiseta incidens is found in almost every part of BC. However, it was not

previously known to occur on Haida Gwaii and other islands along the province’s north

coast. Historical specimens in the Royal BC Museum indicate the presence of Cs.

incidens on Haida Gwaii and other islands along the north coast; an additional specimen

collected in July 2019 by M. Willie confirms the presence of this species in the area

(Table 1). Its range may also extend northwards into the Alexander Archipelago.

Coquillettidia perturbans (Walker). The cattail mosquito, Coquillettidia perturbans

(Walker), is a univoltine mosquito that breeds in shallow bodies of water, such as

swamps, marshes, or shallow lakes, with abundant emergent vegetation and a layer of

soft mud or peat at the bottom (Carpenter and LaCasse 1955; Wood et al. 1979; Belton

1983). Larvae have a highly modified siphon to attach to, and obtain oxygen from, the

roots of emergent aquatic plants (Wood et al. 1979). This species overwinters anchored to

these roots, submerged in mud (Carpenter and LaCasse 1955; Wood et al. 1979).

Coquillettidia perturbans adults possess bands of pale scales on the tarsi, a band of pale

scales in the middle of the tibia, and a mix of large, triangular, black and white scales on

the wing veins (Carpenter and LaCasse 1955; Wood et al. 1979; Belton 1983). Adults

have been reported to feed on nectar from flowers of goldenrod (Solidago spp.)

(Asteraceae), yarrow (Asclepias millefolium), milkweed (Asclepias spp.), dogbane

(Apocynum spp.) (Apocynaceae), and more (Sandholm and Price 1962; Grimstad and

DeFoliart 1974).

Coquillettidia perturbans is aggressive, mammalophilic, and a vector of West Nile

virus (WNV) and eastern equine encephalitis virus (EEEV) (Turell et a/. 2005). It has

often been described as having a southerly distribution (Wood et al. 1979; Belton 1983);

however, it has previously been recorded as far north as Fort Nelson, BC (Poirier and

Berry 2011).

During the July 2019 Yukon Bioblitz, DP collected several adult Cg. perturbans

females attempting to blood-feed. The specimens were from two locations in the southern

Yukon Territory and one location in northern BC (Table 1).

The collections from Upper Liard and Watson Lake represent the first records of this

species and genus in the Yukon Territory and are more than 100 km north and 300 km

west of Fort Nelson, previously the most northerly known collection record for Cg.

perturbans (Poirier and Berry 2011). Both locations were shallow bodies of water with

ample emergent vegetation, including variegated pond-lily, Nuphar_ variegate

(Nymphaeaceae), water horse-tail, Equisetum fluviatile (Equisetaceae), and water

smartweed, Persicaria amphibia (Polygonaceae), and were within the boreal cordillera

ecozone (Canadian Council on Ecological Areas 1996).

74 J. ENTOMOL. Soc. BRIT. COLUMBIA 117, DECEMBER 2020

The collection site at Coal River, BC, was in the forest beside the Alaska Highway,

but because many oxbow lakes are present in the area, we believe our specimens may

have originated from these.

ACKNOWLEDGEMENTS

We thank the Regional District of Fraser-Fort George for funding the 2004 mosquito

trapping program, the Royal BC Museum and the Beaty Biodiversity Museum for access

to their specimens, Taryn Brown, Chris Stinson, Megan Willie, and all who have

collected mosquito specimens referred to in this manuscript. We also thank the organisers

of the Yukon Bioblitz for this fantastic event.

REFERENCES

Belton, E.M. and Belton, P. 1990. A review of mosquito collecting in the Yukon. Journal of the

Entomological Society of British Columbia, 87: 35-37.

Belton, P. 1983. The Mosquitoes of British Columbia. British Columbia Provincial Museum, Victoria,

Canada.

Canadian Council on Ecological Areas. 1996. A perspective on Canada’s ecosystems. CCEA Occasional

Papers, 14. 95 pp.

Carpenter, S. and LaCasse, W. 1955. Mosquitoes of North America (North of Mexico). University of

California Press, Berkeley.

Darsie, R.F.J. and Ward, R.A. 2005. Identification and Geographical Distribution of the Mosquitoes of

North America, North of Mexico. University Press of Florida.

Grimstad, P.R. and DeFoliart, G.R. 1974. Nectar sources of Wisconsin mosquitoes. Journal of Medical

Entomology, 11: 331-341.

Hamer, G.L., Kitron, U.D., Brawn, J.D., Loss, $.R., Ruiz, M.O., Goldberg, T.L., and E.D. Walker. 2008.

Culex pipiens (Diptera: Culicidae): a bridge vector of West Nile virus to humans. Journal of

Medical Entomology, 45: 125-128.

Mullen, G. 1971. The occurrence of Aedes decticus (Diptera: Culicidae) in Central New York. Mosquito

News, 31: 106-109.

Peach, D.A.H. 2017. First record of Aedes (Ochlerotatus) spencerii (Theobald) (Diptera: Culicidae) from

the Yukon. Journal of the Entomological Society of British Columbia, 114: 65-67.

Peach, D.A.H. 2018a. First Record of Culex tarsalis Coquillett (Diptera: Culicidae) in the Yukon. Journal

of the Entomological Society of British Columbia, 115: 123-1235.

Peach, D.A.H. 2018b. An updated list of the mosquitoes of British Columbia with distribution notes.

Journal of the Entomological Society of British Columbia, 115: 126-129.

Peach, D.A.H. and Gries, G. 2020. Mosquito phytophagy — sources exploited, ecological function, and

evolutionary transition to haematophagy. Entomologia Experimentalis et Applicata, 168: 120-136.

Peyton, E.L. 1956. Biology of the Pacific Coast tree hole mosquito Aedes varipalpus (Coq.). Mosquito

News, 16: 220-224. :

Poirier, L.M. and Berry, K.E. 2011. New distribution information for Coquillettidia perturbans (Walker)

(Diptera, Culicidae) in northern British Columbia, Canada. Journal of Vector Ecology, 36: 461-463.

Sandholm, H.A. and Price, R.D. 1962. Field observations on the nectar feeding habits of some Minnesota

mosquitoes. Mosquito News, 22: 346-349.

Smith, M.E. 1952. A new northern Aedes mosquito, with notes on its close ally, Aedes diantaeus H., D.,

& K. (Diptera: Culicidae). Bulletin of the Brooklyn Entomological Society, 47: 19-28, 29-40.

Thielman, A., and Hunter, F. 2007. Photographic key to the adult female mosquitoes (Diptera: Culicidae)

of Canada. Canadian Journal of Arthropod Identification, 4: 1-117.

Turell, M.J., Dohm, D.J., Sardelis, M.R., O’Guinn, M.L., Andreadis, T.G., and Blow, J.A. 2005. An

Update on the Potential of North American Mosquitoes (Diptera: Culicidae) to Transmit West Nile

Virus. Journal of Medical Entomology, 42: 57-62.

Westwood, A.R., Surgeoner, G.A., and Helson, B.V. 1983. Survival of spring Aedes spp. mosquito

(Diptera: Culicidae) larvae in ice-covered pools. The Canadian Entomologist, 115: 195-197.

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Mosquitoes of Canada - Diptera: Culicidae. Research Branch, Agriculture Canada, Ottawa, Canada.

J. ENTOMOL. SOc. BRIT. COLUMBIA 117, DECEMBER 2020 is)

OBITUARY

Leland Medley Humble

(3 November 1951 — 4 August 2020)

Leland Medley Humble passed away peacefully at home on 4 August 2020 with his

family by his side. Lee was born 3 November 1951 in Dawson Creek, British Columbia,

and was raised in Nelson, a beautiful town nestled in the Selkirk Mountains in eastern

BC. Here, where nature was at his doorstep, Lee developed an appreciation of the

outdoors at an early age. Following high school, he worked as a heavy equipment

operator for the Canadian Pacific Railroad; trains remained a passion for Lee for the rest

of his life. Lee attended Selkirk College in Nelson and completed his undergraduate

training in biology at the University of Victoria in 1977. It was here that he started his

life-long fascination with insects and, under the guidance of Richard Ring, completed his

PhD on insect cold tolerance with his dissertation on “Life histories and overwintering

strategies of some arctic sawflies and their hymenopterous parasitoids”. His graduate

research took him to the Canadian high Arctic for several summers. He had many stories

to tell of his adventures in the north: remote camps, exciting flights in small planes,

encounters with large animals and memorable interactions with locals and other

researchers.

These were busy years for Lee; at the same time as being a full time PhD student, he

was the father of a young family and had a job with Dave Gillespie at Agriculture Canada

studying parasites and hyperparasites of the European winter moth, Operophtera

brumata L. In 1985 a position for a forest entomologist came open at the Pacific Forestry

Centre with the Canadian Forest Service in Victoria. John Borden, then a professor at

Simon Fraser University, inspired by a research talk that he heard Lee give, encouraged

him to apply for the job. So began a 35-year career of scientific discovery, collaboration

and fun; Lee often remarked at his good fortune to be paid for enjoying his hobby! Lee

was hired to provide entomology support to the federal Forest Insect Disease Survey

(FIDS), specifically for the BC & Yukon Region. This involved rearing and identifying

forest insects collected in the annual surveys, overseeing the insectary and Pacific

Forestry Centre Arthropod (PFCA) collection and providing general diagnostic services;

and pursuing research interests in what time remained. One of these areas of research

examined the biodiversity of arthropod communities in the tree canopies of temperate

76 J. ENTOMOL. Soc. BRIT. COLUMBIA 117, DECEMBER 2020

rain forests; work with Neville Winchester and Richard Ring and as a component of

the Montane Alternative Silviculture Systems (MASS) project.

Lee was an active participant in the British Columbia Plant Protection Advisory

Council (BCPPAC) and, in addition to chairing the forest pest committee and serving on

the Regional Emergency Action Coordination Team (REACT), he was actively involved

in the science and politics of gypsy moth control. He provided advice regarding the

European gypsy moth, Lymantria dispar dispar L. and the Asian gypsy moth, L. dispar

asiatica, L. dispar japonica, L. umbrosa, L. albescens, and L. postalba, found in North

America, associated with cargo ships in 1991. Lee helped develop a DNA-based

diagnostic test to differentiate the European from the Asian sub-species and in the

process set up a high security quarantine room at the Pacific Forestry Centre. In the early

1990s, his Asian gypsy moth work took him to the Russian Far East where he worked on

an international collaboration addressing moth attraction to light at shipping ports. This

led to adventures in the forests of Siberia and opened his eyes to the significance of the

international movement of forest pests. |

A few years later, in 1996, two important events dramatically affected Lee’s career:

the FIDS program was ended and the Asian longhorned beetle, Anoplophora

glabripennis (Motschulsky), was found in Brooklyn, New York. Lee now focused most

of his time on alien invasive species; identifying established alien species, developing

surveillance and diagnostic tools, studying pest movement pathways and finding practical

mitigation opportunities. -

He kept his research relevant to real issues by working closely with the Canadian

Food Inspection Agency (CFIA), the US Animal and Plant Health Inspection Service

(APHIS), and the US Forest Service as well as international organizations such as the

International Union of Forest Research Organizations (IUFRO), the International

Forestry Quarantine Research Group (IFQRG) and the North American Plant Protection

Organization (NAPPO). Lee carried out joint research projects on forest insect detection

systems in China with scientists at the Chinese Academy of Forestry and was recognized

as an associate research scientist at the Jilin Provincial Academy of Forestry Science. His

expertise in insect rearing techniques provided key data to support the development and

refinement of the international wood packaging standard, ISPM 15. Lee also continued

his work on preventing the introduction and establishment of Asian gypsy moth by

providing his scientific expertise during discussions with Asian countries impacted by the

implementation of the North American phytosanitary standard — Guidelines for

Regulating the Movement of Vessels from Areas Infested with the Asian Gypsy Moth

(NAPPO RSPM 33).

Lee worked throughout his career on locating and identifying predators of hemlock

woolly adelgid, Adelges tsugae Annand. In the early 2000s he identified Laracobius

nigrinus Fender as a biological control agent for use in eastern North America and more

recently he located and collected hemlock woolly adelgid for further work on new

biocontrol agents with graduate students and colleagues in Canada and the USA.

Lee was also keenly interested in the curation of insect collections and contributed to

and modernized the collection at the Pacific Forestry Centre. In addition to the physical

collection, he helped build DNA reference libraries for many groups of insects including

bark and woodborers and contributed through his own collections and those of his PhD

student, Jeremy deWaard, to the Barcode of Life project with Paul Hebert.

In the final major project of his career, Lee circled back to some of the skills he

learned in graduate school, thermotolerance of insects. But this time, instead of cold, he

built a device to very precisely measure the high temperatures required to kill insects.

This elegant piece of equipment, now known as the Humble Water Bath, hand-crafted by

Lee, is a critical tool that will be used to change global trade regulations in years to come.

Lee Humble was a great teacher and mentor. He was an adjunct professor at the

University of British Columbia and served on the Master’s and PhD committees of

numerous graduate students including Ashley Lamb, Gabriella M. Zilahi-Balogh, Jeremy

J. ENTOMOL. SOC. BRIT. COLUMBIA 117, DECEMBER 2020 7]

deWaard, Susanne Kuhnholz, Cynthia Broberg, Stacey Wilkerson, Sepideh

Massoumi-Alamouti and Eveline Stokkink. He also mentored many students through the

co-op program, many of whom came back year after year and were so inspired that they

became entomologists themselves. His true classroom, however, was in the forest where

he knew plants and insects and how they interacted. He was always looking for

ecological connections, formulating theories and planning new experiments. Those who

were lucky enough to work with him in the field experienced Lee in his element. ,

Lee was the recipient of many well deserved awards including the Commemorative

Medal for the 125th Anniversary of the Confederation of Canada, for service to Canada

(1992), the Canadian Forest Service Merit Award for Team Achievement (1999), Natural

Resources Canada (NRCan) Department Merit Award (1999), the Head of the Public

Service Award for Excellence in Policy (1999), Outstanding Foreign Expert Award from

Jilin Province, PR China (2001), Ontario Federal Council, Leadership through

Collaboration Award (2004), the Canadian Forest Service Achievement Award (2012),

Lifetime Achievement Award, Professional Pest Management Association of British

Columbia (2012), NRCan Departmental Achievement Award (2013).

In addition to his passion for entomology, Lee was a skilled woodworker,

photographer and wildlife enthusiast. Many specialized pieces of lab equipment were

meticulously designed and crafted out of wood, plastic or metal by him. He was

brimming with novel ideas, full of knowledge of entomology, botany, ecology, forestry,

and had an unstoppable enthusiasm for science. He was a true renaissance scientist.

Lee Humble was an exceptional scientist who inspired his colleagues with his

passion for science and made significant contributions to forestry and entomology. John

Borden, his mentor and long-time colleague and friend, fittingly said, “An invasive insect

lurking in British Columbia’s forests will not regret the passing of Lee Humble but all of

us who knew him certainly will’.

Meghan Noseworthy and Eric Allen

Pacific Forestry Centre, Victoria

This tribute was originally published in the Bulletin of the Entomological Society of

Canada, Volume 52(4) (December 2020). We thank the Editor of the Bulletin for granting

permission to publish this article.

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NOTICE TO CONTRIBUTORS

The Journal of the Entomological Society of British Columbia is published online as submissions

are accepted. The JESBC provides immediate open access to its content on the principle that

making research freely available to the public supports a greater global exchange of

knowledge. Manuscripts dealing with all facets of the study of arthropods will be considered for

publication. Submissions may be from regions beyond British Columbia and the surrounding

jurisdictions provided that content is applicable or of interest to a regional audience. Review and

forum articles are encouraged. Authors need not be members of the Society. Manuscripts are peer-

reviewed, a process that takes about six weeks.

Submissions. The JESBC accepts only electronic submissions via the journal homepage: http://

journal.entsocbc.ca/. Style and format guidelines, open access fees, and other information for

authors can be found at the journal homepage.

Scientific notes. Scientific notes are an acceptable format for short reports. They must be two

journal pages maximum, about four manuscript pages. Scientific Notes do not use traditional

section headings, and the term "Scientific Note" precedes the title. A short abstract may be included

if desired. Notes are peer- reviewed in the same manner as regular submissions.

Natural History & Observations. Natural history observations are short — typically about two

pages maximum — pieces outlining entomological observations such as (but not exclusively) insect

outbreaks, population collapses, observations in unexpected locations, novel behaviors, etc. The

purpose of these papers will be to record potentially important phenomena — that may otherwise be

overlooked — for the use of future researchers. Papers of this sort were common in many of the

earlier years of the journal, and we feel that they still have a place, particularly as we move further

into the Anthropocene. Natural History and Observations pieces will be subject to peer review.

While not necessarily required, pieces that supply photographic, video, audio, two-witness,

voucher specimens, and/or other rigorous evidence of the observation will be more likely to be

accepted. Photographs will be included in the journal. Video, audio, or other such files should be

deposited in a DOI-based repository (such as figshare, Dryad, or an institutional repository) with

the DOI noted in the written piece. Voucher specimens should be deposited in recognized musuems

with a notation to that effect in the text. The JESBC encourages entomologists and other naturalists

working in British Columbia and the surrounding jurisdictions to consider submitting their

observations as part of the long term record of our regional entomological natural history.

Review and forum articles — Please submit ideas for review or forum articles for consideration to

the editor at journal@entsocbc.ca. Reviews should provide comprehensive, referenced coverage of

current and emerging scientific thought on entomological subjects. Forum articles of about 1000

words in length should provide opinions, backed by fact, on topics of interest to entomologists and

to the general public. Both review and forum submissions are only published in the journal

following full and rigorous peer review.

Electronic reprints. As an open access journal, article reprints from all issues of the journal can be

downloaded as PDFs from the journal homepage.

Back issues. Electronic back issues are available online free-of-charge.

Membership in the Society is open to anyone with an interest in entomology. Dues are $30 per

year for domestic memberships, $15 for students, and $40 for international memberships. Members

receive electronic copies of Boreus (the newsletter of the Society), and when published, occasional

papers. Membership and membership renewals can be purchased at: http://entsocbc.ca/

membership/.

wii

Entomological Society of British Columbia

Volume 117 December 2020 ISSN#007 1-0733

Directors of the Entomological Society of British Columbia 2016-2017 .................. 2

The balsam bark weevil, Pissodes striatulus (Coleoptera: Curculionidae): life history

and occurrence in southern British Colambia ..6042 2... is cece eons cede ccd oe 3

Beetles in the city: ground beetles (Coleoptera: Carabidae) in Coquitlam, British

Columbia as indicators of human disturbance .........: 2.6.5... eta c cc enecneg ser cecsenessee —-20

Effects of trail pheromone purity, dose, and type of placement on recruiting European

fire ants, Myrmica rubra, to toed Dats... 26 o) ca ee ie vee ee 31

Geographic range and seasonal occurrence in British Columbia of two exotic ambrosia

beetles as determined by semiochemical-based trapping ...................cccce cee eee ee ees 42

Andrena (Melandrena) cyanura Cockerell (Hymenoptera: Apoidea, Andrenidae), a valid

North American Species foe ee 49

SCIENTIFIC NOTES

First record of the Palearctic seed bug Metopoplax fuscinervis Stal (Hemiptera:

Oxycarenidae) in Nort America ..0055.05... 60

NATURAL HISTORY AND OBSERVATIONS

Plecoptera from the Crooked River, British Columbia .......................000ceees oe, 64

New distribution records and range extensions of mosquitoes (Diptera: Culicidae) in

British Columbia and the Yukon Termiloty «....22555 4004.6 ee 69

OBITUARY: Leland Medley Humble (3 November 1951 - 4 August 2020)

NOTICE TO CONT Re ae ur ee Inside Back

Cover