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2015-2016

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Department of Natural History, Royal Ontario Museum

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University of Guelph Ridgetown Campus

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JESO Volume 147, 2016

First Canadian record of Atherigona reversura

JESO Volume 147, 2016

FIRST CANADIAN RECORD OF THE BERMUDA GRASS STEM

MAGGOT, ATHERIGONA REVERSURA (DIPTERA: MUSCIDAE)

J. SAVAGE

Department of Biological Sciences, Bishop’s University,

Sherbrooke, Quebec, Canada JIM 1Z7

email, jsavage@ubishops.ca

Scientific Note

The large muscid genus Atherigona Rondani (Diptera) includes over 230 species,

most of which are found in the tropical and subtropical regions of the Old World (Pont and

Magpayo 1995). The group is divided into two subgenera: Acritochaeta Grimshaw, which

contains mostly saprophagous taxa, and Atherigona Rondani, whose phytophagous larvae

are shoot-flies that feed on a variety of wild and cultivated grasses (Poaceae) (Pont and

Magpayo 1995).

Only two species of Atherigona are currently found in North America. The

cosmopolitan Atherigona orientalis Schiner (Diptera: Muscidae) has long been known from

the southern United States (Malloch 1921; Huckett 1936) and has not, so far, been recorded

in Canada. Atherigona reversura Villeneuve (Diptera: Muscidae), a widespread species in

the Oriental, Australasian and Palaearctic regions (Pont and Magpayo 1995), is a much

more recent introduction: it was documented from the United States less than a decade ago

(Hudson 2010) and is recorded here for the first time from Canada. The species was found

to rapidly infest and damage Bermuda grass, Cynodon dactylon (Linnaeus) Persoon, from

hayfields, pastures and turf in the southern United States (Baxter et al 2014), a habit that

has resulted in the recent attribution of the common name Bermuda grass stem maggot

(BSM).

The BSM is a small yellowish fly measuring 3.0-3.5 mm with a remarkably

angular dichoptic head in both sexes and long antennae (Fig. 1A). It can be separated from

A. orientalis and other members of the subgenus Acritochaeta Grimshaw based on the

presence of very short basal lateral setae on the scutellum (these are almost as long as

subbasal lateral setae in A. orientalis), 2-3 rows of presutural acrostical setae (4-5 rows

in A. orientalis ), and a clubbed palpus in the male (Fig. 1A) (male palpus elongate in A.

orientalis). The combination of yellow palps (Fig. 1A), a bicoloured frontal vitta (Fig. IB),

and a moderately ornamented fore tarsus in the male (Figs. 95-96 of Pont and Magpayo

1995) will distinguish A. reversura from other species belonging to the subgenus A therigona

sensu stricto. The immature stages have been extensively described by Grzywacz et al.

(2013). In addition to Bermuda grass, A. reversura has been reared from a variety of host-

plants (see Pont and Magpayo 1995 for complete list) including some cultivated in Canada

such as maize, Zea mays Linnaeus, and, only recently, sweet sorghum, Sorghum hicolor

(Linnaeus) Moench, (Thivierge et al. 2015).

Published March 2016

Savage

JESO Volume 147, 2016

The presence of the BSM in Southern Ontario represents new Canadian records at

both the generic and species levels. While this species was first noticed in the United States

of America due to the damage it caused to Bermuda grass, the Canadian specimens were

discovered through an ongoing project on the DNA barcoding of the Muscidae of Canada

based on a large database of COI Muscidae sequences from all over Canada (Savage et

al. 2015). Eight sequences from that data set belonged to the Barcode Index Number

(Ratnasingham and Hebert 2013) AAN8579 and were assigned to the genus Atherigona

by the automated identification engine of the Barcode of Life Data System (BOLD,

Ratnasingham and Hebert 2007). Upon further examination and genitalic dissections of

the barcoded specimens, the material was identified as A. reversura (2 males, 6 females).

All Canadian records reported here (Table 1) are from Ontario, with the oldest dating back

to 2010. Collection details and individual sequences for the specimens listed here can be

retrieved from BOLD in the public dataset: Atherigona of Canada (dx.doi.org/10.5883/DS-

ATOC).

Additional North American specimens of A. reversura housed in the Biodiversity

Institute of Ontario (Guelph, Ontario) were also seen in the course of this work, including

material from Llorida and California, as well as one male and two females collected in late

September 2010 from Barnstable County, Massachusetts, representing the most northern

record in the United States to date for the BSM. All Canadian specimens are adults collected

by Malaise trap and therefore no information related to host-plant is currently available.

It is not possible at present to determine if the Canadian records reported here

represent multiple punctual introductions of the BSM rather than the establishment of the

species in Canada and no information is available on the cold-hardiness of the species.

LIGURE 1: Atherigona reversura , male. A. habitus, lateral view; B. head, anterodorsal

view.

First Canadian record of Atherigona reversura

JESO Volume 147, 2016

Bermuda grass is considered an exotic weed in Canada, where it is found in Ontario and

British Columbia (Plants of Canada Database 2011). However, since A. reversura is capable

of completing its development in a wide range of other hosts, including some that are

cultivated in Canada, the distribution of this species should be monitored in the future.

Table 1. Collection date, locality, number and sex of specimens and collection repository

for specimens of Atherigona reversura from Canada; all localities are from Ontario. BIO

= Biodiversity Institute of Ontario.

Date

Locality

Lat/Long

Specimen # Collection repository

05.ix.2010

Leeds and Grenville, Elizabethtown-Kitley

44.621, -75.773

1(3

BIO, general collection

19.ix.2010

Haldimond-Dunn Townline, Windy Bluff

42.861, -79.703

1(3,2$

BIO, research collection

of M. A. Smith

21.xi.2010

Haldimond-Dunn Townline, Windy Bluff

42.861, -79.703

BIO, research collection

of M. A. Smith

12.ix.2012

Point Pelee National Park

41.939, -82.516

BIO, general collection

28.ix.2014

Walkerton, Sacred Heart High School

44.127,-81.144

BIO, general collection

28.ix.2014

London, Jack Chambers Public School

43.030, -81.271

BIO, general collection

Acknowledgements

This species was detected as a consequence of a large-scale DNA barcoding

program that was sponsored by the Government of Canada through Genome Canada and

the Ontario Genomics Institute. Specimens were collected and sequenced by staff at the

Biodiversity Institute of Ontario; those from Lake Erie shoreline (Windy Bluff) were

collected with the permissions of the Judd Family using funds from an NSERC Discovery

Grant to M. Alex Smith.

I particularly thank Paul Hebert and M. Alex Smith for access to the specimens and

sequences, Jeremy deWaard for overseeing the collection program, and Valerie Levesque-

Beaudin for preparing the specimens and dataset. I also thank Michael Richardson and

Veronique Bellavance from Bishop’s University (BU) for providing digital images of

specimens. Laboratory space was provided by BU and funding was provided in part by an

NSERC Discovery Grant to J. Savage.

References

Baxter, L.L., Hancock, D.W., and Hudson, W.G. 2014. The Bermudagrass Stem Maggot

{Atherigona reversura Villeneuve): A review of current knowledge. Forage and

Grazinglands 12: 1-8. doi:10.2134/FG-2013-0049-RV

Plants of Canada Database 2011. [online] Available from http://www.plantsofcanada.info.

gc.ca/ [accessed 28 August 2015]

Savage

JESO Volume 147, 2016

Grzywacz, A., Pape, T., Hudson, W.G., and Gomez, S. 2013. Morphology of immature

stages of Atherigona reversura (Diptera: Muscidae), with notes on the recent

invasion of North America. Journal of Natural History 47: 1055-1067. doi:10.10

80/00222933.2012.742244

Huckett, H.C. 1936. A revision of the connectant forms between coenosian and limnophorine

genera occurring in North America (Diptera, Muscidae). Journal of the New York

Entomological Society 44: 187-222.

Hudson, W. 2010. New exotic invasive fly found damaging Bermudagrass forage crops in

Georgia. The University of Georgia College of Agricultural and Environmental

Sciences, Athens (GA).

http ://apps. caes .uga. edu/impactstatements/index. cfm?referenceInterface=IMPAC

T_STATEMENT&subInterface=detail_main&PK_ID=3278

[accessed 22 February 2016]

Malloch, J.R. 1921. A synopsis of the genera of the anthomyiid subfamily Coenosiinae

(Diptera). Entomological News 32: 106-107.

Pont, A.C. and Magpayo, F.R. 1995. Muscid shoot-flies of the Philippine Islands (Diptera:

Muscidae, genus Atherigona Rondani). Bulletin of Entomological Research

Supplement Series 3:1-123.

Ratnasingham S. and Hebert, P.D.N. 2007. BOLD: The Barcode of Life Data System

(www.barcodinglife.org). Molecular Ecology Notes 7: 355-364.

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. doi: 10.1371/

j ournal. pone.0066213

Savage, J., Hebert, P.D.N., and Levesque-Beaudin, V. 2015. The Muscidae of Canada:

towards a complete DNA barcode reference library. Pp. 277 In Adamowicz, S.J.

(ed). Scientific abstracts from the 6th International Barcode of Life Conference /

Resumes scientifiques du 6 e congres international Barcode of Life. Genome 58:

163-303. doi: 10.1139/gen-2015-0087

Thivierge, M.-N., Chantigny, M.H., Belanger, G, Seguin, P, Bertrand, A., and Vanasse, A.

2015. Response to nitrogen of sweet pearl millet and sweet sorghum grown for

ethanol in eastern Canada. BioEnergy Research 8: 807-820.

A modified technique for rearing wood boring insects

JESO Volume 147, 2016

A MODIFIED TECHNIQUE FOR REARING WOOD

BORING INSECTS PERMITS VISUALIZATION OF LARVAL

DEVELOPMENT

T. SHARMA 1 *, J. DEDES 1 , C. J. K. MACQUARRIE 1

‘Natural Resources Canada Canadian Forest Service, Great Lakes Forestry Centre,

1219 Queen Street East, Sault Ste. Marie, Ontario, Canada P6A 6T3

email, triptisharmaca@gmail.com

Abstract J. ent. Soc. Ont. 147: 7-14

Studying the larvae of wood boring insects is difficult because this stage of

the life cycle is spent hidden from the eyes of researchers. Previous studies

have used sections of bark and phloem sandwiched between pieces of glass

or plastic to permit observation within the host substrate. The development of

artificial diets for bark and wood boring insects has allowed the development

of new techniques to study these insects under more consistent conditions.

We modified an artificial diet in order to replicate the phloem sandwich

technique by using a tortilla press to prepare sheets of artificial diet and

then tested this technique by rearing larvae of the emerald ash borer, Agrilus

planipennis Fairmaire (Coleoptera: Buprestidae). We reared neonates and

older larvae extracted from infested trees on the diet sandwiches and found

rates of establishment and survivorship were higher for older larvae extracted

from trees than for neonates. Our technique improves upon traditional phloem

sandwiches by allowing all larvae to be reared under the same conditions

and permitting the observation of larval behaviour even when the larva is

submerged in the diet.

Published December 2016

Introduction

Studying the ecology and behavior of bark and wood boring beetle larvae is

difficult because this part of the life cycle is hidden from researchers. Bedard (1933) devised

a solution to this problem by placing the inner bark of Douglas fir, Pseudotsuga menziesi

(Mirb.) Franco (Pinaceae), between two glass slides held in place with elastic bands to study

the Douglas-fir beetle Dendroctonuspseudotsugae (Hopkins) (Coleoptera: Curculionidae).

Since then the technique has been modified, extended, and come to be known by the

name ‘bark sandwiches’ or ‘phloem sandwiches’ (Yu and Tsao 1967; Beanlands 1966;

Wermelinger and Seifert 1998).

* Author to whom all correspondence should be addressed.

Sharma et al.

JESO Volume 147, 2016

Emerald ash borer (EAB), Agrilusplanipennis Fairmaire (Coleoptera: Buprestidae),

is a highly destructive pest of ash, Fraxinus (Oleaceae). Studying the larval ecology of

EAB has meant either removing larvae from infested ash (Wang et al. 2010), which kills

the insect, or observing larvae in standing trees (Duan et al. 2010), which is logistically

difficult. The recent development of an artificial EAB diet (Keena et al. 2015) has allowed

new ways to study EAB under controlled conditions. We developed a “diet sandwich”

method inspired by the phloem sandwich method using sheets of artificial diet sandwiched

between two microscope slides. We then tested our ability to visualize EAB larvae in the

diet as they developed and quantified survivorship and overall development time.

Materials and Methods

All diet preparation was done under sterile conditions in the diet kitchen at Natural

Resources Canada Canadian Forest Service, Great Lakes Forestry Centre (GLFC), Insect

Production Unit, Sault Ste. Marie, Ontario, Canada. For these experiments we used the

ingredient list from “diet #3” from Keena et al. (2015) (Table 1) with minor revisions. We

used a cutter mixer (model HCM450, Hobart Canada, North York, Ontario) rather than a

steam kettle to mix the ingredients, which resulted in more homogenous diet product. We

then used a 23 cm diameter tortilla press (model Victoria-85008, Imusa USA, Doral, Florida,

U.S.A) to make diet sheets and vacuum sealed the diet sheets for long-term storage.

TABLE 1. Ingredients of emerald ash borer artificial diet #3 from Keena et al. (2015).

Mix

Ingredient(s)

Quantity (g)

Starting base

Agar (Gracilaria spp.)

220

Sodium bicarbonate

Dry mix # 1

Casein (edible grade)

200

Sucrose

300

Wesson salt mix without iron

Sorbic acid

Calcium propionate

Methyl para benzene

(Methyl paraben)

Potato starch

200

Dry mix #2

Soy flour

500

Wet mix

Cholesterol

Wheat germ oil

Vitamins and fibre

Choline chloride

Vitamin A acetate beadlets

Bioserv#F8128 (vitamin mix)

Alphacel

1,500

A modified technique for rearing wood boring insects

JESO Volume 147, 2016

Preparation of diet and diet sandwiches

1. Boil autoclaved reverse-osmosis water in a steam kettle and add sodium bicarbonate

to neutralize the pH of the water.

2. Add agar with continuous stirring by spatula, so that no clumps form and heat until

boiling.

3. Transfer the agar solution to a cutter mixer and add dry mix #1 (Table 1) and mix

at 1140 rpm for 1 min.

4. Quickly add soy flour and mix at 1140 rpm for 1 min.

5. Add wet mix (Table 1) and mix at 1140 rpm for 3 min.

6. Allow the diet to cool until the temperature reaches 50C (this step is necessary as

it prevents heat degrading the vitamins added in step 7).

7. Add dry mix #2 (Table 1) and mix at 1140 rpm for 1 min.

8. Add 750 g of Alphacel and mix it 1140 rpm for 1 min, then add remaining 750 g

and mix again for 1 min.

The product from the above procedure should be the consistency of bread dough. To prepare

the diet sheets we used the following procedure:

1. With gloved hands, break the large mass into 5 to 10 small balls, each approx. 10

cm in diameter.

2. Place the diet balls into closed plastic bags to prevent drying.

3. Spread 0.5 ml of wheat germ oil on a 60 x 30 cm piece of wax paper and place one

end of the wax paper (oil side up) on a tortilla press.

4. Make small balls (4-4.5 cm diameter) from the large diet balls and place one on

tortilla press on top of the wax paper.

5. Fold the remaining wax paper over the ball and flatten the press until the ball

reaches the desired thickness (3-6 mm) (Fig. 1A).

6. Vacuum seal the diet sheets in plastic bags (10 sheets/bag) and store at 4C. The

sheets are good for up to 6 months.

Finally, to prepare the diet sandwiches we used the following procedure:

1. Remove 1 diet sheet from the plastic bag and place one glass slide (7.5 x 5.1 x 0.1

cm; No. 2957, Erie Scientific Company, Portsmouth, New Hampshire) on top of

the sheet.

2. With a scalpel, cut the diet sheet using the glass slide as a template.

3. Place the diet sheet between two glass slides to form a sandwich.

4. Wrap the sandwich in clear plastic wrap.

5. Prepare the desired number of sandwiches and store at 4C until needed.

Introduction of larva on diet sandwich

Larvae were introduced to the diet sandwiches by placing them into a small notch

cut into the upper surface of the diet sandwich using a scalpel (Fig. IB). Other methods

suggest introducing EAB to the diet as eggs and allowing the neonates to hatch directly into

the diet (Keena et al. 2015). However, we found that placing larvae allowed us to confirm

that viable individuals are being used in our experiments. Individual diet sandwiches were

then placed upright in small boxes (15 x 7 x 6.5 cm) inside an opaque plastic box (35 x 25

Sharma et al.

JESO Volume 147, 2016

x 18 cm). A 1-2 cm deep layer of water was put in the opaque box to maintain humidity,

and the small plastic boxes were placed on a plastic grid in the bottom of the opaque box to

prevent them from being submerged. All larvae were reared in an environmental chamber

(24C; 65% RH; 16L:8D). The developing larvae occasionally needed to be moved to a new

diet sandwich (e.g., when the diet dried out, or when the larvae consumed a large portion of

the diet) using the technique described above.

EAB larvae reared on an artificial diet typically stop feeding after the 4 th instar

and require a chill period to develop further (Keena et al. 2015). This part of the life cycle

can be recognized by morphological changes in larval features such as the larvae reducing

in length, becoming whiter and more opaque, the thoracic segments becoming narrower

and shorter than the abdominal segments, and most notably, it stops feeding altogether and

bends itself into a J-shaped configuration. The rearing temperature was gradually decreased

to 16C for 7 d then to 10C for 7 d and finally to 4C for 84 d. After 84 d, larvae were removed

from the diet sandwiches and transferred individually into wells of a 6-well culture plate

FIGURE 1: (A) Tortilla press used to press sheet of artificial emerald ash borer diet, (B)

into which a larva is placed via a slit cut into the upper surface of the diet sandwich. (C)

Implanted larvae create serpentine galleries in the artificial diet sandwich, (D) which can

be used to follow the progress of development, including molting.

A modified technique for rearing wood boring insects

JESO Volume 147, 2016

(Becton Dickinson Labware, Franklin Lakes, NJ. USA 07417-1886) for pupation and adult

development. A moist cotton pad covered with 8 to 10 layers of paper towel was placed in

the bottom of each well to provide a rough substrate to help with molting. Well plates were

stored in the same opaque plastic boxes used to hold the diet sandwiches and maintained

at the same rearing conditions (24C; 65% RH; 16L: 8D) until pupation. During this time

the cotton pad and paper towels were re-moistened as necessary to maintain humidity and

prevent fungal growth.

To demonstrate that diet sandwiches are a viable rearing technique, we evaluated

the success of EAB development under the conditions described above. We tested the success

of EAB larvae from two sources: those hatched from eggs (neonates, < 24 h old) and those

extracted from infested ash trees (extracted larvae). We obtained EAB eggs from a colony

maintained at GLFC which were allowed to hatch and neonate larvae were transferred to the

diet sandwiches. Extracted larvae were either removed from sections of ash trees obtained

from infested ash stands or removed from standing infested trees. Larvae were collected

individually into well plates or petri plates with moistened filter paper, then immediately

placed onto diet sandwiches. We placed 83 l st -4 th instar extracted larvae, determined by

measuring head capsule width (Chamorro et al. 2012) and 135 neonates on individual diet

sandwiches. The moisture content of fresh diet and diet that had been stored for 6 months

was determined using a moisture analyzer (Model MF-50, A&D Company Ltd, Toshima-

ku, Tokyo, Japan).

Results

EAB larvae began tunneling, and visible galleries could be seen 2 or 3 d after

being placed on diet (Fig. 1C); however, some larvae took up to 30 d to begin feeding.

Approximately half (52%) of the infested larvae established on the diet. We observed slightly

higher establishment rates among extracted larvae (54%) than neonates (50%) (Table 2).

However, 13.3% (n = 18) of the larvae infested as neonates escaped the diet and perished.

Of the larvae infested as neonates, 6 reached the 4 th instar, of which 4 were placed into the

chill period but none converted into pupa. Twenty of the extracted larvae were placed into

the chill period, of which 3 pupated and 2 emerged into adults. Comparing development

times we found that, for larvae infested as neonates, development time to 3 rd instar was

(mean ± 1 s.d.) 73 ± 40 d (n = 9) and to the 4 th instar was 195 ± 66 d (n = 6). For extracted

larvae, development time to 3 rd instar was 174 ± 98 d (n = 9) and to fourth instar was 171

± 36 d (n = 8). For comparison, previous studies observed development times for male and

female EAB of 176 ± 11 d and 145 ± 9 d, respectively, at 25 ± 2C (Keena et al. 2015).

We found a small decrease in moisture in the diet during the 6 month storage

period (59% for fresh diet vs. 54% for 6 month old diet).

Discussion

We found that EAB larvae infested as neonates and extracted from ash adopted and

fed on artificial diet sandwiches, and their development was visible through the glass slide.

We also observed establishment (52%) on artificial diet sandwiches comparable to rates of

Sharma et al.

JESO Volume 147, 2016

68% and 70% in earlier studies that used a version of the same artificial diet (Chen et al.

2011; Keena et al. 2015). The development times we observed were slightly longer than

previous studies; however, these studies reared their insects at higher temperatures.

There are a number of factors that influence the success of insects feeding on an

artificial diet. For instance, both high and low moisture content in artificial diets containing

ash phloem increases the mortality of larvae (Chen et al. 2011). The optimal moisture content

of EAB diet is 50 (Keena et al. 2015) or 60% (Chen et al. 2011), while the moisture content

of our freshly made diet was 59% (54 after storage). By contrast, in living ash, the moisture

content of phloem varies from 44% to 52% (Hill et al. 2012). This may in part explain the

different performance of larvae in our experiment. In future it may be prudent to desiccate

the diet before constructing sandwiches to more closely replicate moisture conditions in ash

phloem. Introducing live larvae may also be too severe a trauma compared to introducing

larvae from hatched eggs (Keena et al. 2015). Thus, while our method should result in

a higher number of live larvae being placed on diet it may also reduce survivorship. We

also lost a significant number of larvae due to them escaping the sandwich. This was not

anticipated as larvae feeding in cut ash logs rarely escape the phloem, and we anticipated the

same behavior would occur here. To resolve this, it may be necessary to use an impenetrable

barrier around the edge of the diet sandwich to prevent the insect from escaping. Increasing

the thickness of the diet sheet may also increase the success rate of EAB on diet entering the

J-stage and successfully overwintering.

Diet sandwiches permit the observation of developing larvae in a consistent

environment overtime, without disturbing the larvae’s feeding process. Therefore we argue

the method has use as a research tool for the study of larval wood borer behaviour and

ecology (Fig. ID). For instance, we have used the technique to determine developmental

rates for EAB larvae (CJKM unpublished) and observed how galleries changed in size and

conformation over time. We propose that the technique could be applied to the study of

new insecticides or the role of phytochemicals in ash defensive response (Chakraborty et

TABLE 2. Percent establishment of emerald ash borer larvae from two sources implanted

into diet sandwiches, and the final stage reached.

Source

Stage started

% established

(n)

Number reaching stage

(n)

Chill

Pupa

Reared

Neonates

135

49.6 (67)

Extracted

57.9(11)

62.9(17)

37.5 (9)

61.5(8)

Total

54.2 (45)

A modified technique for rearing wood boring insects

JESO Volume 147, 2016

al. 2014). The advantage of diet sandwiches over traditional phloem sandwiches is that the

method is not limited by the number of bark discs that can be collected, nor is it influenced

by variability among trees that are used to obtain bark and phloem samples. Other studies

have used crumbled artificial diet for bioassays in micro-centrifuge tubes to test for the effect

of phytochemicals on larval development (Whitehill et al. 2014). However, that method

does not simulate the feeding environment the insect normally experiences inside the tree,

which could influence both the behaviour and subsequent survivorship of the insect. Using

the diet sandwich technique we have also observed that even if the larva is immersed in the

diet, it still can be observed by placing the sandwich in front of bright light. This permits

continuous observation of larvae when they are submerged, a feature that is not usually

possible with traditional bark and phloem sandwiches or crumbled diet in tubes. Moreover,

we see no barrier to the technique being adapted to the study of other bark and wood boring

beetles that can also be reared on artificial diets (Gould et al. 2004; Gindin et al. 2009).

Acknowledgements

We thank K. Boissoneau, S. Dumas, S. Fiset, Z. Li, F. McReady, G. Mick, R.

Scharbach, and J. St. Amour for their assistance with the experiment, and M. Keena for

advice in preparing the EAB diet. We also thank the five anonymous reviewers for comments

that improved this manuscript.

References

Beanlands, G. E. 1966. A laboratory-rearing method for observing adult bark beetles and

their developing broods. The Canadian Entomologist 98: 412—414. doi: 10.4039/

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Exclusion fencing inhibits beetle activity-density in broccoli JESO Volume 147, 2016

EXCLUSION FENCING INHIBITS EARLY-SEASON BEETLE

(COLEOPTERA) ACTIVITY-DENSITY IN BROCCOLI

J. M. RENKEMA 1 *, B. G. EVANS 1 , C. HOUSE 2 , R. H. HALLETT 2

‘Gulf Coast Research and Education Center, University of Florida,

14625 County Rd. 672, Balm, Florida, USA 33598

email, justin.renkema@ufl.edu

Abstract J. ent. Soc. Ont. 147: 15-28

Exclusion fencing represents a potentially useful management tool for key

insect pests in broccoli but may also affect other invertebrates that have

important roles in agroecosystems. Because beetles (Coleoptera) are generally

abundant and diverse in agriculture and some species (i.e., members of the

Carabidae and Staphylinidae) are important for biological control, pitfall traps

were used in this study to compare beetle communities during late spring and

early summer 2013 in fenced, unfenced and control plots of broccoli. Control

plots were separated from fenced and unfenced plots to determine whether

fencing increased captures in adjacent unfenced plots. Early on, fewer beetles

(total and for most functional trait categories) were captured in fenced

plots, but as the season progressed captures were similar among plot types.

There was little evidence that fencing increased beetle diversity or activity

density in adjacent unfenced plots, and later in the season some ground

beetle species were instead strongly associated with control plots. Pitfall

traps captured relatively high numbers of crucifer flea beetle, Phyllotreta

cruciferae Goeze. Most captures were in unfenced and control plots early in

the season, suggesting that fencing was effective in keeping this pest away

from broccoli. Overall, fencing could limit or delay surface-active predatory

beetles from accessing broccoli fields, having a negative effect on biological

control services. However, since many beetles eventually permeated fencing,

modifications to the fencing design may allow beetles entry, restrict exit, and

allow increase of beetle communities to improve biological control services

in fenced areas.

Published December 2016

* Author to whom all correspondence should be addressed.

2 School of Environmental Sciences, University of Guelph, 50 Stone Road West, Guelph

ON, Canada NIG 2W1

Renkema et al.

JESO Volume 147, 2016

Introduction

Development of new methods for use in integrated pest management is needed in

order to reduce reliance on insecticides and provide solutions to organic growers (Bomford

et al. 2000a). Natural enemies may contribute to control, and cultural practices, such as

crop rotation, field sanitation, removal of alternate host plants/weeds, trap cropping, and

manipulation of planting dates, can be used to reduce pest populations (Acheampong and

Stark 2004; Ito et al. 2005; Corlay et al. 2007; Hemachandra et al. 2007; Chen et al. 2009;

Hummel et al. 2010; Jia-Ying et al. 2010). Exclusion fencing (i.e., panels of fine mesh

surrounding a crop) has been used as a physical practice to reduce pest infestations in

horticultural crops (Vernon and McGregor 1999; Bomford et al. 2000a, b; Wyss and Daniel

2004), but impacts on associated entomofauna have not been examined.

While exclusion fencing can be effective in reducing pest damage in cole crop

production (Vernon and Mackenzie 1998), the purpose of this study was to assess fencing

impacts on diversity and activity density of surface-dwelling beetles. Many of these beetles

are generalist predators (Coleoptera: Carabidae, Staphylinidae) known to prey on important

crop pests (Collins et al. 2002; Hajek et al. 2007; Renkema et al. 2014). However, within

these two beetle families and others common in agriculture (eg., Coleoptera: Scarabaeidae,

Curculionidae, Elateridae), many species may have other functional roles as detritivores,

fungivores, phytophages, or rhizophages (Clough et al. 2007; Renkema et al. 2016). Thus,

grouping beetles by feeding guild across taxa may provide a better prediction of expected

ecosystem services (e.g., predation of crop pests, nutrient cycling) than the use of taxonomic

groupings alone.

Exclusion fencing studies have shown that since many agricultural pests fly at

their highest density ca. 30 cm above ground, fences taller than 30 cm are necessary to

block pest movement into cropped areas (Tuttle et al. 1988; Vernon et al. 1989; Vernon

and Mackenzie 1998; Vernon and McGregor 1999; Bomford et al. 2000a, b; Wyss and

Daniel 2004). Fences create shelter and provide calm air, with the result that there may

be 2 to 7 times more insects on the leeward compared to windward side (Pasek 1988). A

windbreak effect of fencing increases the potential for high insect numbers along perimeters

(Bomford et al. 2000b). Fencing and exclusion barriers have been most commonly used

in conjunction with pitfall traps to reduce densities of surface-dwelling predators in

experimental areas, resulting in greater pest populations (Chiverton 1987; Holland 1998;

Menalled et al. 1999). In a few cases, creating “egress” boundaries to prevent generalist

predator emigration reduced sentinel prey or pest population build-up compared to field

areas without boundaries (Menalled et al. 1999; O’Neal et al. 2005).

Many surface-dwelling beetles, particularly ground beetles (Carabidae), use field

margins and may immigrate to crop land during spring and summer (Woodcock et al. 2005;

Werling and Gratton 2008). Therefore, beetle captures should be lower in fenced than

unfenced plots, with the degree of difference dependent on how well fences exclude beetles

and the proportion of field-resident beetles in total numbers captured. The distribution of

beetles between fenced and unfenced plots may change as the season progresses and beetles

find their way into fenced areas. Finally, since ground beetles aggregate at edges (Hansen

Exclusion fencing inhibits beetle activity-density in broccoli JESO Volume 147, 2016

and New 2005), fencing may serve to increase beetle numbers in unfenced plots that are in

close proximity to fenced plots compared to unfenced plots at a greater distance.

Materials and Methods

Study Site

This study was conducted in a broccoli, Brassica oleracea L. Italica group, field

(24 x 30 m), at the University of Guelph Elora Research Station, established to examine

effects of variety and fencing on swede midge, Contarinia nasturti (Kieffer), infestation

(Evans and Hallett, unpublished data). In the fencing efficacy experiment, there were four

treatment combinations (i.e., two broccoli varieties, ‘Bay Meadows’ and ‘Windsor’, either

fenced or unfenced) replicated five times and arranged in a randomized complete block

design, with replicates as blocks. The field was divided into twenty 3x5m plots separated

by a 2 m border, with each plot consisting of four rows of 12 broccoli plants, with 95 cm

between row spacing and 45 cm between plant spacing. Broccoli seedlings were grown

from seed (Stokes Seed Co., Thorold, Ontario, Canada) in a greenhouse at the University

of Guelph, using 96-cell flat trays and potting media (Pro-mix: 75-85% sphagnum peat

moss, coarse perlite, vermiculite; Premier Horticulture Ltd., Dorval, Quebec, Canada),

and fertilized once per week (10-52-10 plant starter fertilizer, Plant Products Co. Ltd.,

Brampton, Ontario, Canada). Eight-week-old seedlings were transplanted into the field 7-9

May 2013. Plots were hand-weeded prior to the initial transplant of seedlings and again

on 20 June 2013. No insecticides were applied during the experiment. In order to account

for potential effects of fencing on neighbouring unfenced plots, eight control plots of the

same dimensions were planted in a broccoli field adjacent to the fenced/unfenced plots with

seedlings of the same age, using the same protocols.

One day after transplanting, four 1.5 m tall ‘no-see-um’ mesh panels (Telstar

Window Service Ltd., Agassiz, BC, Canada) were arranged around each fenced plot in a 5.5

x 5.5 m square. The bottom edge of the mesh fencing was buried under ~10 cm of soil, such

that no gap occurred between the fence and the soil surface. Fence panels had 25 cm mesh

overhangs angled 45° outward and downward to prevent insects from flying over the top

edge of the fencing (Pats and Vernon 1999; Vernon and McGregor 1999).

Sampling Protocol

Epigeic beetles were sampled 17 June - 19 July 2013. One of the five blocks (two

fenced and two unfenced plots) and two of the eight control plots were randomly selected

for sampling each week. Each block and control plot was sampled once, so that every

fenced and unfenced plot in the study site was sampled, and so that sampling results in the

previous week(s) would not affect subsequent sampling results by trapping out resident

beetles in each plot. The two control plots sampled in the first week were resampled in the

fifth week. Plots were sampled for five consecutive days using pitfall traps constructed

from clear plastic 10 cm diameter deli containers (500 ml. Solo Cup Company, Lake Forest,

IL), filled one-quarter with a 50% propylene glycol solution, as described in Brunke et al.

(2014). Pitfall traps were placed in pairs within the plots, with one trap placed randomly

in an outer row of plants and a second trap placed randomly in an inner row. Traps were

Renkema et al.

JESO Volume 147, 2016

dug into the soil so that the top edge was level with the soil surface, and were protected

from rain with a plastic cover, supported by wire pegs and positioned 5-10 cm above the

trap. Contact of trap edges to the soil surface was maintained every second day. Traps were

removed after 5 days, and contents were washed with water through a 425 pm sieve (Fisher

Scientific, Ottawa, Ontario) and stored in 70% ethanol until further processing. All beetles

were pinned, most were identified to genus or species with existing literature (Bousquet

2010; Brunke et al. 2012) and assistance from taxonomic experts (Steve Paiero, University

of Guelph; Adam Brunke, Agriculture and Agri-Food Canada), and all are stored at the

School of Environmental Sciences, University of Guelph.

Data analysis

Beetle captures were summed over both pitfall traps per plot, and activity density

and species richness were compared among plot types using individual-based rarefaction,

extrapolating rarefaction curves to 500 individuals. To evaluate intraseasonal differences in

beetle diversity, the first two sampling weeks were designated as the early period and the

final three weeks as the late period. Rarefied species richness and Simpson’s diversity index

values (1/D) were calculated per plot per period, extrapolating species richness estimates

for each plot to the plot with the greatest captures overall. Beetle genera or species were

assigned to a trophic group using available literature (Good and Giller 1991; Farochelle

and Fariviere 2003; Clough et al. 2007; Fundgren 2009; Brunke et al. 2012). Analysis

of variance (ANOVA) was used to compare beetle activity density, rarefied extrapolated

species richness, and diversity and activity density of each major functional group, among

the three treatments. Sampling dates were included as random effects. Residuals were

checked for normality of error variance, and activity density data for each functional group

were square-root transformed. Back-transformed means and 95% confidence intervals are

shown. Estimates 9.1.0 was used to generate rarefaction estimates and Simpson’s diversity

index values (Colwell 2013), and JMP® software (SAS 2013) was used for ANOVA, with

a = 0.05.

Unconstrained ordination in CANOCO 5 (terBraak and Smilauer 2012) was used

to examine differences in beetle community composition among plot types. Only species

(log [x + 1] transformed) with five or more total captures were used. Principle component

analysis (PCA) was performed with data centered by species. A biplot with scaling of scores

focused on inter-species correlations was generated with the 15 species having the largest

relative weight in the analysis.

Results

A total of 1,170 beetles were captured in this study, belonging to 11 families and 42

genera/species. A few beetles, 1.5% of the total, were identified only to family or subfamily

(Table 1). About half as many beetles were captured in fenced compared to unfenced and

control plots (Fig. 1), but species richness was not affected, as 95% CIs overlapped (data not

shown). Fower captures in fenced plots were most apparent during the first two sampling

weeks (Fig. 2). Indeed, beetle captures were greater in control than fenced plots during the

Exclusion fencing inhibits beetle activity-density in broccoli JESO Volume 147, 2016

TABLE 1. Beetles captured in fenced, unfenced and control broccoli plots at Elora

Research Station, June 17 - July 19, 2013. Percent calculated from total number of beetles

captured.

Family

Genus/species

No. (%)

Functional Group

Anthicidae

Anthicus Paykull sp. 1

10(0.8)

Scavenger

Anthicus sp. 2

1(0.1)

Scavenger

Carabidae

Agonum muelleri (Herbst)

2 (0.2)

Predator

Amara aenea (DeGeer)

4(0.3)

Omnivore

Amara obesa (Say)

3 (0.3)

Omnivore

Amara Bonelli sp.

1(0.1)

Omnivore

Anisodactylus sanctaecrucis (F.)

65 (5.6)

Omnivore

Bembidion quadrimaculatum Say

34 (2.9)

Predator

Bembidion Latreille sp.

4 (0.3)

Predator

Bradycellus Erichson sp.

1(0.1)

Predator

Clivina fossor (L.)

9 (0.8)

Predator

Elaphropus anceps (LeConte)

1(0.1)

Predator

Elaphropus (Motschoulsky) sp.

2 (0.2)

Predator

Harpalus affinis (Schrank)

158(13.5)

Omnivore

Harpalus Latreille sp.

6 (0.5)

Omnivore

Loricera pilicornis (F.)

3 (0.3)

Predator

Poecilus lucublandus (Say)

40 (3.4)

Predator

Pterostichus melanarius (Illiger)

196(16.8)

Predator

Stenolophus comma (F.)

116(9.9)

Omnivore

Stenolophus ochropezus (Say)

1(0.1)

Omnivore

species 1

5 (0.4)

species 2

1(0.1)

Chrysomelidae

Chaetocnema Stephens sp. 1

6(0.5)

Phytophage/Rhizophage

Chaetocnema sp. 2

4(0.3)

Phytophage/Rhizophage

Phyllotreta cruciferae (Goeze)

147(12.6)

Phytophage/Rhizophage

Phyllotreta striolata F.

10 (0.8)

Phytophage/Rhizophage

Coccinellidae

CoccineUa septempunctata (L.)

1(0.1)

Predator

Cryptophagidae

Atomaria Stephens sp.

40 (3.4)

Mycetophage

Curculionidae

Cleonis pigra (Scopoli)

1(0.1)

Phytophage/Rhizophage

Glocianus Reitter sp.

1(0.1)

Phytophage/Rhizophage

Sitona Germar sp.

2 (0.2)

Phytophage/Rhizophage

Sphenophorus Schoenherr sp.

24 (2.0)

Phytophage/Rhizophage

species 1

2 (0.2)

Dermestidae

Attagenus Latreille sp.

2 (0.2)

Trogoderma Dejean sp.

1(0.1)

Elateridae

Aeolus mellillus (Say)

21 (1.8)

Phytophage/Rhizophage

Hemicrepidius Germar sp.

1(0.1)

Phytophage/Rhizophage

Hypolithus Eschscholtz sp.

1(0.1)

Phytophage/Rhizophage

species 1

1(0.1)

species 2

1(0.1)

Nitidulidae

Glischrochilus fasciatus (Olivier)

2 (0.2)

Scavenger

Glischrochilus quadrisignatus (Say)

80 (6.8)

Scavenger

Carpophilus Stephens sp.

6(0.5)

Scavenger

Scarabaeidae

Aphodius Hellwig sp.

1(0.1)

Phytophage/Rhizophage

Renkema et al. JESO Volume 147, 2016

TABLE 1 continued...

Family

Genus/species

No. (%)

Functional Group

Staphylinidae

Aleochara Gravenhorst sp.

20(1.7)

Predator

Anotylus insecatus (Gravenhorst)

16(1.4)

Scavenger

Anotylus rugosus (F.)

5 (0.4)

Scavenger

Dinaraea angustala (Gyllenhal)

30 (2.6)

Predator

Drusilla canaliculata F.

32 (2.7)

Predator

Lathrobium Gravenhorst sp. 1

1(0.1)

Predator

Lathrobium sp. 2

3 (0.3)

Predator

Philonthus Stephens sp.

7(0.6)

Predator

Quedius Stephens sp.

2 (0.2)

Predator

Rugilus Leach sp.

4(0.3)

Predator

Scopaeus Erichson sp.

4(0.3)

Predator

Tachinus corticinus Gravenhorst

16(1.4)

Predator

Tachyporus nitidulus (F.)

1 (0.1)

Mycetophage

Tachyporus Gravenhorst sp.

1 (0.1)

Mycetophage

Aleocharine sp. 1

4(0.3)

Predator

Aleocharine sp. 2

1 (0.1)

Predator

Aleocharine sp. 3

1 (0.1)

Predator

species 1

1 (0.1)

TOTAL

1170

FIGURE 1. Rarefied estimates of beetle species richness in fenced or unfenced broccoli

plots or control plots at Elora Research Station, early summer 2013. Rarefaction indicated

with solid lines and extrapolation with dashed lines. Confidence intervals were removed

for visual clarity.

Exclusion fencing inhibits beetle activity-density in broccoli JESO Volume 147, 2016

■Control

-Unfenced

-Fenced

FIGURE 2. Total numbers of beetles captured in pitfall traps in fenced and unfenced

broccoli plots and control plots at Elora Research Station, 2013.

early period, but differences among plot types were not recorded in the later period (Table

2, Fig. 3). Generic/species richness and diversity did not vary due to fencing (richness: F 721

= 3.3, P = 0.059; diversity: F 221 = 2.0, P = 0.165), period (richness: F 13 = 0.1, P = 0.803;

diversity: F l% - 0.05, P = 0.837), or the fencing x period interaction (richness: F 22l = 1.1, P

= 0.355; diversity: F 22l = 0.9, P = 0.416).

There were effects of fencing on most trophic groups of beetles (Table 2). Predatory

beetle captures increased in fenced plots from the early to the late trapping periods, whereas

omnivore captures remained lowest in fenced plots throughout the experiment (Fig. 3).

Rhizophage and phytophage (i.e., potential pest) numbers decreased in control and unfenced

plots from the early to the late trapping period, but remained low in fenced plots throughout

the experiment (Fig. 3).

Ordination eigenvalues were 0.223 and 0.168 for the first two axes, respectively,

and explained 47.7% of the cumulative variability in beetle species distribution. None of the

frequently captured species were associated with fenced plots in four of the five sampling

weeks (Fig. 4). All three plot types grouped closely for the sampling week of 1-8 July and

were associated with Bembidion quadimaculatum Say and a Harpalus sp. Three larger,

common predatory species of Carabidae, Harpalus affinis (Schrank), Poecilus lucublandus

(Say), and Pterostichus melanarius (Illiger), tended to associate with control plots during

the later sampling weeks.

Renkema et al.

JESO Volume 147, 2016

TABLE 2. Results of analysis of variance for effects of fencing (control, unfenced, fenced)

and sampling period (early = 17 June - 1 July, late = 2-22 July) on numbers of all beetles

and beetles categorized by trophic designation captured in broccoli plots at Elora Research

Station, 2013.

All beetles Predators Omnivores

rtujiuis

Fencing

6.2

0.008

1.2

0.312

6.7

0.006

Period

1.1

0.373

11.8

0.041

0.2

0.691

Fencing*period

5.8

0.010

3.8

0.039

1.3

0.289

Phytophages +

Rhizophages

Scavengers’

Mycetophages

Fencing

14.2

<0.0001

2.1

0.152

1.3

0.302

Period

12.7

0.038

0.1

0.773

2.5

0.211

Fencing*period

8.8

0.002

4.7

0.020

1.3

0.299

df: Fencing = 2, 21; Period =1,3; Fencing*period = 2, 21

’Fencing*period means not significantly different as indicated by Tukey’s HSD test, a =

0.05

. . :...

JLJ

Unfenced

Early

E±i

1 ; .i

: ■ I I

Control Unfenced Fenced

m Mycetophage

□ Scavenger

E9 Phytophage/Rhizophage

□ Omnivore

■ Predator

FIGE1RE 3. Mean (back-transformed) numbers of beetles categorized by trophic level

captured per week in pitfall traps from broccoli plots that were unfenced or fenced or from

control areas outside plots, at Elora Research Station, 2013. The experiment was divided

into early (June 17-July 1) and late (July 1-22) trapping periods. For total beetles (A, B),

omnivores (a, b) and predators (y, z) different letters indicate significantly different means

using Tukey’s HSD test, a = 0.05. Confidence intervals (95%) are shown only for predators

and phytophages/rhizophages to improve visual clarity.

Exclusion fencing inhibits beetle activity-density in broccoli JESO Volume 147, 2016

FIGURE 4. Principle component analysis biplot of beetle species and plot type for each

sampling week in a broccoli field, at Elora Research Station, 2013. Species are shown by

arrow vectors, empty circles are control plots, empty triangles are unfenced plots and filled

squares are fenced plots. See Table 1 for full species names.

Discussion

The community composition of beetles, particularly Carabidae, captured in this

study were relatively similar to communities in other horticultural and field crops. Harpalus ,

Pterostichus, mdAnisodactylus ground beetles were common in vegetable crops in western

North Carolina, and, in addition, Bembidion spp. were common in eastern England Brassica

crops (Hummel et al. 2002; Eyre et al. 2009). In Ontario sweet potato and carrot fields,

the dominant ground beetle genera were Pterostichus , Harpalus , Bembidion , Poecilus and

Scarites , and these as well as Clivina , Agonum and Tachys were common in Ontario field

crops (maize, soybeans, wheat) (Belaoussoff et al. 2003; Brunke et al. 2009). While direct

comparisons of beetle activity density or diversity between studies are difficult to make,

similarities in predaceous and omnivorous ground beetle genera may mean community

functionality - pest control and weed seed predation - is also relatively similar across crop

types and/or locations.

Using fencing to exclude pests from broccoli had significant effects on beetle

activity density, as fewer beetles were found in fenced compared to unfenced and control

plots early in the study (17 June - 1 July). This result is due to the fact that most surface-

active beetles were likely immigrating from field margins into the establishing crop at this

time (i.e., Werling and Gratton 2008; Eyre et al. 2009), and fences effectively kept them

from walking into plots. Despite influencing beetle abundance, fencing did not significantly

impact generic/species richness or diversity. This result suggests that fencing does not

Renkema et al.

JESO Volume 147, 2016

preferentially exclude particular taxonomic or functional groups of ground-dwelling beetles,

but restricts access equally across beetle types.

We did not expect to capture many foliage-dwelling beetles in pitfall traps, but

Phyllotreta cruciferae Goeze were abundant early in the study (138 early captures of 147

total captures) and comprised the majority (67%) of phytophages/rhizophages. This flea

beetle is a widespread crucifer pest, causing defoliation and reduced yield (Tansey et al.

2008). Row cover can be used to protect young crucifer plants by excluding P. cruciferae

(Andersen et al. 2006), but it also appears that exclusion fencing provides protection from

this pest.

In the later part of the trapping season (2-22 July), beetle activity-density was not

affected by fencing. Captures in control and unfenced plots tended to decrease compared to

the early period, whereas captures in fenced plots tended to increase. This result suggests

that beetles, of which predatory and omnivorous Carabidae and Staphylinidae were a large

majority, moved into the fenced plots through gaps that occasionally opened at seams in

fence panels or between the panels and the ground, or that species capable of flight entered

over the top edge. Satiation can affect ground beetle movement, with hunger motivating

dispersal (Fournier and Loreau 2001). High flea beetle levels outside fenced plots early in

the season may have meant that predatory beetles frequently encountered prey (flea beetles

or other prey) (Renkema et al. 2014), but as flea beetle numbers declined, predatory beetles

may have increased their search activity and found their way into fenced plots.

No differences in beetle captures were found between unfenced and control plots,

despite expecting greater beetle abundance and diversity in unfenced plots due to a barrier

effect of adjacent fences. A tendency for association between control plots and some of the

larger, more frequently captured ground beetle species (i.e., H. affinis , P. lucublandus, Pt.

melanarius) later in the trapping period may be due to weedy vegetation around these plots

or gaps in the fences or at lower edges that were not well maintained. In addition, these plots

were less disturbed by other research activities associated with measuring fencing effects

on pests, and these species may favour less disturbed habitats (Carmona and Landis 1999).

Using exclusion fencing to assist with broccoli pest management has the unintended

consequence of excluding beneficial beetles from fields, or at least delaying their entry.

Thus, biological control services could be reduced early in the season, and prevent natural

enemies from limiting pest population growth (Hajek et al. 2007). In addition, effects of

fencing small plots (this study) on beetle entry may be different than effects at a commercial

scale, where a lower perimeter distance to fenced area ratio may further reduce likelihood

of beetle entry. However, the fact that many beetles eventually permeated the fencing

is promising for later season biological control services, but species-specific effects of

predators on pests will need to be determined, as has been done in other southern Ontario

horticultural crops (Brunke et al. 2009). The feeding ecology of rove beetles should be

determined, particularly of species common in both this study and a previous survey of

southern Ontario soybeans, such as Drusilla canaliculata and Dinaraea angustala (Brunke

et al. 2014). Fencing designs that allow ingress (so that beetles can walk in but not out easily)

and result in retaining abundant beetle populations in fenced fields, leading to greater insect

pest predation and agroecosystem services, should also be investigated.

Exclusion fencing inhibits beetle activity-density in broccoli JESO Volume 147, 2016

Acknowledgements

We thank Jamie Heal, Emily Anderson, Taylor LaPlante, Caitlyn Schwenker, and

the staff at the Elniversity of Guelph Elora Research Station. This project was funded by

an Ontario Ministry of Agriculture, Food and Rural Affairs (OMAFRA) New Directions &

Alternative Renewable Fuels Research Program grant to RHH.

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Mitteilungen der Deutschen Gesellschaftfuer Allgemeine und Angewandte

Entomologie 14: 387-390.

JESO Volume 147, 2016

POSTER AND PRESENTATION ABSTRACTS

ENTOMOLOGICAL SOCIETY OF ONTARIO

ANNUAL GENERAL MEETING

Great Lakes Forestry Centre, Sault Ste. Marie, Ontario, October 14 - 16, 2016

J. ent. Soc. Ont. 147: 29-41

Poster abstracts

Sublethal impacts of pesticide exposure on bumble bees and squash bees, and a wild

pollinator monitoring programme for Ontario

Agar, E., G.L. Baron, M.J.F. Brown, D.S .Chan, R.J. Gill, V.A.A. Jansen, A. Pindar, C.

Rubens, D.A. Stanley, and N.E. Raine

New range records from a headwater in a beef pasture provide evidence of agricultural

streams as important habitat for invertebrates

DeGasparro, S.L. 1 andD.V. Beresford 2

1. Trent University Environmental and Life Sciences Program, 2. Trent University

Departments of Biology and Sustainable Agriculture and Food Systems

Headwater streams contribute to the biodiversity of freshwater systems and

provide important habitats for invertebrates in agricultural landscapes. Compared to higher

order and larger streams, little is known about the ecology of small headwater streams

and the taxa associated with them. We report on 28 invertebrate species found in a small

headwater stream on a beef pasture in central Ontario (Canada) over two years. Two species,

Lepidostoma liba (Ross) (Trichoptera: Lepidostomatidae) and Pericoma albitarsis (Banks)

(Diptera: Psychodidae) are first provincial records for Ontario, and several others are

significant range extensions or gap infills. We discuss the importance of continued sampling

in small, overlooked habitats to better understand species diversity.

Where do the army worm come from? Using stable isotopes to study the migration of

Pseudaletia unipuncta

Doward, K., J.N. McNeil, and K.A. Hobson

In Canada, many lepidopteran pest species, such as the true armyworm {Pseudaletia

unipuncta ), are seasonal migrants. Consequently, as little is known about their origin other

than “they come from the south”, current management strategies are generally based on

pheromone trap catches to estimate the density of immigrants. However, if immigrant

populations come from specific areas year after year, one could potentially estimate the size

of immigrant populations based on overwintering conditions at the source site.

Published December 2016

JESO Volume 147, 2016

The existence of a latitudinal continental spatial hydrologic scale for deuterium

in precipitation (and thus in the plants growing at different sites) provides a means of

determining the natal origin of migrant species. Consequently, I am using hydrogen isotope

ratios (c 2 H) to determine the degree of intra- and inter-year variability in armyworm moths

captured in London, ON. Initially, I reared cohorts of larvae on vegetation ( Hordeum

vulgare ) treated with water having different concentrations of deuterium to establish the

relationship between water d 2 H and wing chitin o 2 H. As there are three periods of armyworm

flight activity (immigrants, residents and emigrants) I am now analysing the wings of field-

collected moths that were captured in different years and at different times during the

seasons.

Our working hypothesis is that within a year, immigrants captured in spring will

have significantly different d 2 H profiles than those in summer and fall, as they will have

fed on host plants growing in locations much further south while residents and emigrant

populations will have fed on local vegetation. The inter-year comparison will allow me to

determine to what extent the origin of immigrants varies from one year to the next.

Range expansion pattern of Carabus granulatus Linnaeus (Coleoptera: Carabidae) in

eastern North America

Fleming, K. 1 , and D. Beresford 2

1. Trent University Environmental and Life Sciences Program, 2. Trent University

Departments of Biology and Sustainable Agriculture and Food Systems

Carabus granulatus Linnaeus (family: Carabidae) is native to Europe. It was first

introduced to North America in 1890 in St. John, New Brunswick. In 2011, C. granulatus

was collected in Moosonee, Ontario, extending the known distribution northward by 200

kilometers. Typically, invasive species follow one of three different types of radial expansion

curves, based on the dispersal characteristics (Shigesada and Kawasaki 1997). Using

published records of previous sightings, we analyzed the expansion rate of C. granulatus

in eastern North America. C. granulatus likely dispersed by diffusion, (Type 1 expansion

curve) through eastern North America, characteristic of other coleopteran invaders.

Urban effects on blow fly species diversity across Canada

Langer, S. 1 , C. Kyle 2 , and D. Beresford 3

1. Trent University Environmental and Life Sciences Program, 2. Trent University Department

of Forensic Science, 3. Trent University Departments of Biology and Sustainable Agriculture

and Food Systems

We tested whether the ecozone an urban area is located in, or the urban area itself

had a greater effect on forensically relevant blow fly (Diptera: Calliphoridae) species

composition across Canada. We mailed bottle traps to 32 collaborating OPP and RCMP

locations across Canada in 2011,2012 and 2013. We found that blow fly species composition

in urban areas across Canada were similar over a range of ecozones.

JESO Volume 147, 2016

Biosurveillance of Alien Forest Enemies (bioSAFE): Pathway analysis and diagnostics

of Asian Longhorned Beetle

Roe, A.D., C. Duff, R. Hamelin, K. Hoover, M. Keena, C. Landry, M. Javal, S. Juan, A.

Roques, and Y. Wu

Efficacy of insecticides for control of brown marmorated stink bug (Halyomorpha

halys Stal) nymphs in Ontario

Scott-Dupree, C. 1 , A. Gradish 1 , K. Hunter 1 , H. Fraser 2 , and T. Gariepy 3

1. University of Guelph, Guelph, ON, 2. OMAFRA, Guelph, ON, 3. AAFC, London, ON.

Halyomorpha halys , the brown marmorated stink bug (BMSB), is an invasive

agricultural and household pest native to East Asia. Identified as established in Ontario

in 2012, its range is expanding into agricultural areas. Sustainable pest management

options must be identified to prevent significant economic loss for Ontario growers. Older,

broad-spectrum insecticides appear to be most effective for managing BMSB but may

be undesirable due to non-target impacts. Therefore, screening of alternative insecticides

are required. Results of laboratory studies using direct contact toxicity tests of novel

insecticides (alone and in combination) on fifth instar nymphs are discussed.

Japanese knotweed Psyllid Aphalara itadori: A biological control agent for invasive

knotweeds

Skuse, T

The psyllid Aphalara itadori has recently been approved for open release to help

control invasive knotweed plants (Fallopia sp.) in Canada. Before widespread releases into

knotweed-infested areas can occur, there is a need to further investigate how these insects

will establish in novel ecosystems. We will present data from ongoing research partnered

with Agriculture and Agri-food Canada about the ecology of a new bug entomologists can

expect to see more of in the near future.

Overwintering energetics and microhabit selection in the western bean cutworm

Turnbull, K.F., J.N. McNeil, and B.J. Sinclair

Overwintering insects risk depleting energy stores at increased temperatures due

to an elevation in metabolic rate. However, insects can conserve energy behaviourally

by selecting a microhabitat that buffers against higher temperatures, or physiologically

through metabolic plasticity. The western bean cutworm ( Striacosta albicosta) appears to

use both strategies; the soil depth of prepupal chambers is a determinant of overwintering

temperatures, and warmer conditions induce a decline in the thermal sensitivity of

metabolism. We demonstrate that soil texture and temperature at the time of burrowing

influence overwintering site selection, which may have implications for overwintering

success.

JESO Volume 147, 2016

Distribution of Syrphidae in northern Ontario

Vezsenyi, K. 1 , J.H. Skevington 2 , W.J. Crins 3 , J. Schaefer 1 , D. Beresford 4

1. Trent University, Environmental and Life Sciences Program, 2. Canadian National

Collection of Insects, Arachnids, and Nematodes, 3. Ministry of Natural Resources and

Forestry, 4. Trent University, Departments of Biology and Sustainable Agriculture and

Food Systems

I report on the distribution of an important pollinator group, hover flies (Syrphidae),

from Ontario’s northern boreal region, and Akimiski Island, Nunavut. Little is known

regarding the distribution of this family in the more remote parts of northern Ontario. To

date, I have identified 49 species from 251 individuals, with about 400 still to be processed.

Fifteen of these are range extensions, and provide new records of rare species. This project

will add to our understanding of the distribution of this important group.

Presentation abstracts

Intra-caste variation in a eusocial sweat bee, Lasioglossum laevissimum

Awde, D

Sweat bee studies typically explore how abiotic and environmental factors

influence female social status but rarely focus on intra-caste variation. In a southern Ontario

Lasioglossum laevissimum population, spring and summer queens were similar in size and

ovarian development but differed in foraging behaviour. Half of workers had some ovarian

development and 20% of workers had substantial ovarian development, suggestive of

queen-like behaviour. These queen-like workers were similar in size to classic workers.

Our results show considerable intra-caste variation, emphasizing the behavioural flexibility

inherent in L. laevissimum castes.

Is competition superior to parasitism for biological control? The case of spotted

wing drosophila ( Drosophila suzukii), Drosophila melanogaster and Pachycrepoideus

vindemmiae

Dancau, T.f T.L.M. Stemberger, P. Clarke, and D.R. Gillespie

1. Carleton University.

Drosophila suzukii and Drosophila melanogaster coexist with overlapping

resource use. When sharing resources in the lab, D. melanogaster outcompetes D. suzukii.

We allowed adult D. suzukii and D. melanogaster females to compete for access to a

common oviposition resource in dyadic and population scale experiments and a closed field

simulation cage experiment with a generalist Drosophila parasitoid, Pachycrepoideous

vindemmiae. Competitor identity was found to significantly affect D. suzukii numbers

over the presence of P. vindemmiae. Competition by D. melanogaster appears to be more

effective as biological control for D. suzukii than a parasitoid in this system, http://dx.doi.

org/10.1080/09583157.2016.1241982

JESO Volume 147, 2016

Effects of cold acclimation on structure and transport function in insect ionoregulatory

tissues

Des Marteaux, L. 1 , and B.J. Sinclair 1

1. University of We stern Ontario

Chill-susceptible insects lose ion and water homeostasis and develop chilling

injuries when exposed to cold, but prior cold acclimation allows insects to maintain

homeostasis to lower temperatures and avoid injury. The mechanisms underlying cold

acclimation are not well-understood, but likely involve modification of cell/tissue structure

and/or epithelial transport function. The insect hindgut and Malpighian tubules (major sites

of ionoregulation) are likely targets for these modifications. In adult Grylluspennsylvanicus

crickets we characterized the effects of cold acclimation on structural and ionoregulatory

gene expression, hindgut cytoskeletal stability, and Malpighian tubule secretion. We found

that cold acclimation alters both active transport function and cell structure, and discuss

how these modifications may prevent chilling injury and loss of homeostasis.

Staphylinid diversity across a Neotropical elevation gradient

Dolson S., E. Loewen 1 , K. Jones 1 , and M.A Smith 1

1. Department of Integrative Biology, University of Guelph, 50 Stone Rd E, Guelph, ON

N1G2W1; dolsons@uoguelph.ca.

Climate change is altering the diversity, abundance, and distribution of natural

populations. Of these populations, those in tropical montane areas are expected to be the

first affected by changing conditions. Studying the effects of abiotic and biotic conditions on

tropical community composition and distribution can provide insight into what community

changes may occur in the future as these factors shift. Elevation gradients provide a useful

model system to study these effects since many conditions covary as elevation increases,

namely temperature (decreasing) and precipitation (increasing). Understanding community

response to changing conditions is particularly important in arthropods as most are

understudied despite their abundance and importance in various ecological populations.

Therefore, I focus my research on defining the staphylinid communities across an elevation

gradient in the Area de Conservacion Guanacaste in Costa Rica and determining how they

are affected by the changing environmental factors. Continuous collections have been made

at 8 sites across a 1500 m elevation gradient since 2008 and to quantify diversity we will use

DNA barcodes and classic taxonomy when available. Investigation into these communities

will not only provide insight into important yet unknown biogeographical patterns of an

understudied taxa but also into how ecological communities may respond to changing

climatic variables.

Evidence for extended parental care in the eastern carpenter bee, Xylocopa virginica

Duff, L

The eastern carpenter bee, Xylocopa virginica has two provisioning flight periods,

one to provision nest-mates from late April to early May and another to provision brood

cells from late May to early July. In St. Catharines, Ontario we observed both female and

JESO Volume 147, 2016

male bees leaving nests in late August and in the beginning of September. During this time,

we made novel observations that worn and unworn females made pollen provisioning trips

to nests. In Ontario, carpenter bees have little time to provision a second brood, so late

summer provisioning flights may be intended as extended parental care.

New pest issues in Ontario horticulture in 2016

Fraser, H

Ontario Ministry of Agriculture, Food and Rural Affairs

Invasive insects have resulted in the disruption of many well-established integrated

pest management programs. In 2016, the European cherry fruit fly, Rhagoletis cerasi (L.)

(Diptera: Tephritidae), was detected at several locations in southern Ontario. The find

represents the first confirmation of this pest in North America. The presence of pepper

weevil, Anthonomus eugenii Cano (Coleoptera: Curculionidae), not though to overwinter

successfully under typical Ontario winters, has resulted in crop loss in both greenhouse and

field peppers. Pepper weevil has the potential to become a community-level pest under

certain conditions. Both pests are discussed in the context of Ontario production.

Status of ambrosia beetles in high-density apple orchards in southern Ontario

Fraser, H. 1 , E. Richard, and C. Scott-Dupree

1. Ontario Ministry of Agriculture, Food and Rural Affairs

Wood-boring ambrosia beetles have been reported as an emerging issue in the

United States where they have caused substantial damage in high density apple orchards

over the last few years. A preliminary survey was conducted to determine the range,

abundance and diversity of ambrosia beetles in 13 high density apple orchards located in

southern Ontario. A survey for ambrosia beetles was conducted in 13 high density apple

orchards. Six species of ambrosia beetle were identified including Anisandrus dispar , A.

sayi , X germanus, Xyleborus obesus , Monarthrum mali , and Xyleborinus saxesenii. Beetle

numbers in 2016 were low relative to those reported in other jurisdictions in 2015.

Discovery of a parasitoid attacking the invasive swede midge in Ontario

Hallett, R.H. 1 , S. McGregor, J. Williams, and J.D. Heal

1. School of Environmental Sciences, University of Guelph.

The invasive alien swede midge, Contarinia nasturtii (Diptera: Cecidomyiidae),

was first recorded from canola fields in Shelburne, ON in 2003. In 2016, weekly plant

collections were made at four canola fields in this area in order to look for the presence of

swede midge parasitoids. A platygastrid was found in all fields over a 6-week period, with

parasitism rates ranging from 4% to 20%. Future studies are planned to learn more about

this parasitoid and its biological control potential against the swede midge.

JESO Volume 147, 2016

Trichoplusia ni (Hiibner) attraction to transgenic Solanum lycopersicum (L): Exploring

transgenic trap crops for the cabbage looper moth

Laur, W. 1 - 2 ,1.M. Scott 2 and J.N. McNeil 1

1. Department of Biology, Western University, London, Ontario, 2. London Research and

Development Centre, Agriculture and Agri-Food Canada

New pest management strategies are required to counter insecticide-resistance in

populations of greenhouse insects, for example Trichoplusia ni (Hiibner), the cabbage looper

moth. One alternative is to use trap crops - planting attractive, disposable plants within the

main crop arrangement to draw pests away from the crop. In this study, transgenic Solanum

lycopersicum (L) tomato was genetically modified to over-express the gene that regulates

carotenoid cleavage dioxygenase (CCD) enzymes, altering production of tomato volatiles.

The objectives of the study are to verify and assess cabbage looper moth olfactory attraction

to volatiles emitted by transgenic tomato lines relative to wild-type.

An investigation of Staphylinidae functional diversity along a neotropical elevation

Loewen, E. 1 , S. Dolson 1 , and M.A. Smith 1

1. Department of Integrative Biology, University of Guelph; eloewen@mail.uoguelph.ca

Staphylinidae is a hyperdiverse and abundant family of Coleoptera found

worldwide. My research focuses on how Staphylinidae functional diversity varies with

environmental changes associated with the elevation gradient (e.g. temperature decreases

with increasing elevation) on Volcan Cacao in the Area de Conservacion Guanacaste (ACG)

in northwestern Costa Rica. Since 2008, multiple samples have been taken at various

elevations (10m above sea level -1500m above sea level) and in the three distinct forest types

(dry forest, rainforest and tropical montane cloud forest (TMCF)) that characterize Volcan

Cacao. Through morphometric measurements and discreet variables such as exoskeleton

texture and colour, I will investigate the changes in Staphylindae functional morphospace

along the elevation gradient and between the forest types. Like many tropical arthropods,

our understanding of staphylinids suffers severe taxonomic impediments due to the relative

scarcity of taxonomists working on them, many unnamed and undescribed species, and the

many described species that may represent multiple separate species. My proposed research

will add to our limited knowledge on neotropical staphylinids and test predictions regarding

how abiotic factors such as temperature and precipitation affect functional measures of

diversity.

Alien (spider) invaders: Enoplognatha ovata wreak havoc on a coastal arthropod

community

McCann, S. 1 , C. Scott, and M.C.B. Andrade

1. Department of Biological Sciences, University of Toronto Scarborough; Department of

Ecology and Evolutionary Biology, University of Toronto.

Enoplognatha ovata (the candy stripe spider) is a theridiid introduced to North

America from Europe, and common on the West Coast. Much-studied because of its striking

colour polymorphism, its ecology has thus far attracted little interest. Here we report on the

JESO Volume 147, 2016

remarkable predation behaviour of this unassuming spider, observed during a longitudinal

study of a field site on Vancouver Island, BC. We show how E. ovata uses theft, piracy, and

strategic web-building to carve a trail of havoc across the sand dunes, with the potential for

significant impact on the native arthropod community, including hymenopterans and other

spiders.

Wing interference patterns (WIPs) - a potential tool for the identification of

Sciomyzidae (Diptera) of eastern Canada and the adjacent U.S. states

Muzzatti, M. 1 , and S. Marshall

1. M. Sc. candidate in Environmental Science, University of Guelph, 254 Cole Rd., Guelph,

ON NIG 3K4; muzzattm@uoguelph.ca.

Wing interference patterns (WIPs) are specific colour patterns developed through

thin film interference that appear on translucent insect wings, and are a potential source

of useful taxonomic data. WIPs were discovered in 88 species across 21 genera of

Sciomyzidae (Diptera) from eastern Canada and adjacent U.S. states. Moderate to high

interspecific variation and low to moderate intraspecific variation of WIPs was discovered,

along with one record of sexual dimorphic WIPs. WIPs of Sciomyzidae are of most value

in a taxonomic identification key as a supplemental taxonomic trait to improve difficult

couplets, such as those that require examination of internal genitalia.

The value of entomology field courses

Otis, G.W

School of Environmental Sciences, University of Guelph; Guelph, Ontario NIG 2W1;

gotis@uoguelph. ca.

Steve Marshall and I began teaching Field Entomology 20 years ago. Many

undergraduate students get little exposure to field activities. Providing them those

opportunities can be transformative. Our course has turned-on many students to the world

of insects and field research, with many now contributing professionally to the discipline of

entomology. Major ingredients for a successful course are keeping students warm, clean,

well-fed, and happy. The sites where the course is taught must be interesting biologically

and hopefully physically appealing. We encourage live-in courses at field stations, not

weekday-daytime camps, where students become absorbed in course activities.

Effect of gut-associated yeasts on Drosophila melanogaster performance

Jimenez Padilla, Y. 1 , M-A Lachance 1 , and B.J. Sinclair 1

1. Department of Biology, University of Western Ontario.

Yeasts are an important part of the Drosophila diet due to their nutritional value,

and as such, yeasts have the potential to affect phenotype. Furthermore, living yeast might

also interact with the fly as its host in ways we have not yet discovered. While yeasts are

an integral part of the fly gut community, most studies have focused on bacteria alone.

However, Lachancea kluyveri, a yeast commonly associated with Drosophila in nature,

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affects fly physiology, and the magnitude of the effect is dependent on the yeast being alive

or dead.

Insecticide impacts on bumblebees: from colony founding to pollination services

Raine, N.E. 1 ’ 2 , G.L. Baron 2 , M.J.F. Brown 2 , and DA. Stanley 23

1. School of Environmental Sciences, University of Guelph, Guelph, Ontario, NIG 2W1,

Canada, 2. School of Biological Sciences, Royal Holloway University of London, Egham,

Surrey, TW20 OEX, UK, 3. Botany and Plant Science, School of Natural Sciences and Ryan

Institute, National University of Ireland, Galway, Ireland.

Recent concern over global pollinator declines has led to considerable research on

pesticide impacts. Here we report results from a series of studies examining to what extent

field-realistic insecticide exposure can lead to significant sublethal impacts on individual

behaviour (e.g. reduced queen colony founding success and impaired worker learning and

foraging), colony function (e.g. effects on growth rates and forager recruitment), and the

critical ecosystem services bumblebees provide to crops and wild plants. Taken together

these effects could have widespread implications for the stability of wild pollinator

populations, sustainable production of pollinator limited crops, and maintaining wild plant

biodiversity.

Experimentally induced alloparental care in a small carpenter bee

Richards, M. and V. Lewis.

Alloparenting, in which adults help to raise non-descendant offspring, is the

hallmark of both cooperatively breeding and eusocial animal groups. In the small carpenter

bee, Ceratina calcarata, mothers sometimes produce very small daughters, reminiscent

of eusocial workers, suggesting retention of a complex social trait, alloparenting, in

this secondarily solitary bee species. Experimental removal of mothers does induce the

smallest daughters to feed their siblings, but natural orphaning always results in brood

predation. Thus the necessity for maternal care precludes the possibility of alloparenting

and explain why this bee has reverted to solitary behaviour.

The mating dynamics of western black widows: new insights from field observations

Scott, C. 1 , S. McCann, and M.C.B. Andrade

1. Department of Biological Sciences, University of Toronto Scarborough; Department of

Ecology and Evolutionary Biology, University of Toronto.

The western black widow, Latrodectus hesperus , has been the subject of several

experimental studies of sexual behaviour and communication, yet our knowledge of its

natural history remains incomplete. We conducted a six-month longitudinal study of a

dense population of L. hesperus on Vancouver Island, BC. We report new information about

phenology, and how this intersects with changing modes of sexual selection, including the

timing of mate-searching and mating, the frequency of mate-guarding, and the intensity of

male-male competition. These data are essential for designing new behavioural experiments

and interpreting results of earlier studies.

JESO Volume 147, 2016

Evaluating the interaction between buckwheat Fagopyrum esculentum (Polygonaceae)

and wireworm (Coleoptera: Elateridae) species

Scott, I.M. 1 , Y.L. Bohorquez Ruis 12 and J.N. McNeil 2

1. London Research and Development Centre, Agriculture and Agri-Food Canada, 2.

Department of Biology, Western University, London, Ontario

Click beetle larvae or wireworms are considered pests due to the damage caused

by feeding on the root systems of agricultural crops. Field studies in Atlantic Canada

using buckwheat in rotation with potato resulted in a reduction of Agriotes spp. wireworm

populations. Microplot trials with buckwheat and an Ontario species, Limonius agonus

(Say), demonstrated a negative effect on larvae during a 3 week interaction. Laboratory

soil olfactometer experiments found no evidence that Agriotes sputator L. were deterred

by buckwheat at germinating, branching and flowering stages relative to red spring wheat

(Triticum spp.) and island barley ( Hordeum vulgare). In the greenhouse, a 3 week, no choice

feeding assay determined no difference in weight and mortality of A. sputator larvae when

fed buckwheat or barley. However, in contrast to barley, the wireworm herbivory did not

affect buckwheat growth, suggesting that buckwheat roots may produce anti-feedants.

Comparison of complexity and intelligence among cynipid gall wasps and wild roses

Shorthouse, J

Professor Emeritus - Department of Biology, Laurentian University; Sudbury ON P3E

2C6.

One approach to understanding the life history strategies of phytophagous insects

and their host plants is to compare their relative complexity and even their intelligence.

Although all zoologists are comfortable discussing the relative complexity of their study

animals, most are likely uneasy when considering insects and plants as intelligent. Here

I use gall wasps of the genus Diplolepis (Family Cynipidae) and their host shrub roses of

the genus Rosa (Family Rosaceae), to question which is more biologically complex and

intelligent, gall wasps represented by both larvae and adults, or their host shrubs?

Because adult cynipids have a nervous system and fly to locate oviposition sites,

whereas shrubs are sessile and without a nervous system, most biologists likely agree that

cynipids are more complex than plants. Cynipid larvae are also considered complex because

they control plant growth causing their larvae to be encased by protective layers of high

quality plant food in new organ-like structures. However, cynipids spend most of their lives

as either larvae or prepupae and only two weeks as pupae and adults. Immature stages are

immobile and have no opportunity to make decisions.

Wild roses with their modular structures and clonal growth forms are also complex

organisms as they search for optimum habitats, assess signals and make decisions as they

slowly grow the shoots and roots of their ramets. Roses have growing points both above and

below the ground where they receive and assess information on light, temperature, touch,

soil structure, insect attack, microbes, gravity, gradients of water, nutrients, minerals and

toxins, and chemical signals from other plants.

Rose shrubs respond to more signals than cynipids suggesting a transduction

JESO Volume 147, 2016

network of substantial complexity. Even though the leaves, buds, flowers, stems, and roots

of a rose bush operate with some degree of independence, they are integrated to form a

complex organism and they accomplish this without overall controlling organs or a nervous

system. They plan their growth trajectory and there is evidence that each perceived signal

presents different problems that requires intelligent mastery.

A common definition of intelligence in plants includes an intrinsic ability to process

information from both abiotic and biotic stimuli that allows optimal decisions about future

activities in a given environment in order to solve problems. Roots of plants, for example,

investigate, search, survey, examine, and discover.

While we debate the legitimacy of using terms such as learning, memory, decision¬

making and intelligence when studying plants, and the ways in which they react to insect

attack, it is becoming easier to accept that some plants exhibit brainy behaviour in the absence

of brains. There is even evidence that some plants can become primed after repetitive insults

of heat, herbivores, drought, cold, etc. such that they respond more quickly in the future to

these conditions. In other words, they exhibit memory.

If intelligence reflects the capacity to solve problems, then a case can be made

that shrub roses are more intelligent than cynipids because they react to a wider range of

problems both above and below the soil surface. It appears that wild roses have become

information processing entities of such complexity, integration, and adaptive competence

that they rival those of many advanced animals and current electronic systems. We are

reminded that after Charles Darwin studied the sensory capabilities of plant roots, he asked

us to think of plants as a kind of up-side down animal with the main sensory organs and

‘brain’ on the bottom deep underground and its sexual organs on top.

Assessment of the role of neonicotinoid seed treatments to manage early season corn

pests

Smith, J. 1 , T. Baute 2 , and A. Schaafsma 1

1. Department of Plant Agriculture, University of Guelph Ridgetown Campus, 2. Ontario

Ministry of Agriculture, Food, & Rural Affairs.

A four year study is underway in Ontario, Canada to evaluate the utility of

neonicotinoid insecticide seed treatments in a corn and soybean production system.

The objectives of this study are to 1) determine the key early season insect pests and

their distribution in Ontario corn and soybeans, 2) develop early season insect pest risk

assessments tools, and 3) to measure the economic impact of neonicotinoid seed treatments

for early season pest control in these crops. Approximately 150 replicated strip plots were

conducted in 2014 and 2015 on cooperator’s farms comparing corn and soybeans treated

with a fungicide or fungicide + neonicotinoid seed treatment. Early season foliar and soil

insect pest presence were assessed using destructive sampling and bait trapping methods to

determine species composition and damage levels. Plant populations were measured, plots

were harvested by the cooperators and yield data were reported to the researchers. The key

wireworm and grub species found in Ontario corn and soybeans will be reported along with

the results of wireworm bait trap method comparisons. Early indications of monitoring

for soil pests in the fall or spring suggest that applying insecticides based on the observed

presence of the pest will be challenging. The results of this study will also provide context

JESO Volume 147, 2016

to the overall discussion of the value of neonicotinoid seed treatments in corn and soybeans

in Ontario, Canada.

When ants get up; they sure get brown

Smith, M.A

Department of Integrative Biology, University of Guelph.

Using a long-term collection of ants from an neotropical elevation gradient in

Area de Conservacion Guanacaste (ACG) in northwestern Costa Rica we have tested the

prediction that in cool environments ectotherms should be, darker to maximize heat gain

and larger to minimize heat loss. We found that assemblages from higher elevation sites

(cloud forests) were darker than rain or dry forests. Mid-elevation sites were characterised

by the greatest range of lightness values. Furthermore, we found that cloud forest ant

assemblages are becoming significantly lighter with time, suggesting the arrival of

downslope assemblage members coincident with the increasing dryness and brightness that

is affecting contemporary cloud forests.

Evaluating the efficacy of pure and hybrid populations of a biological control agent

SzucSjM. 1 , U. Schaffner 2 , P. Salerno 3 , J. Littlefield 4 , B. Teller 5 , K. Shea 6 , and R. Hufbauer 1

1. Department ofBioagricultural Sciences and Pest Management, Colorado State University,

Fort Collins, CO 80523-1177, USA, 2. CABI - Europe, Switzerland, Rue des Grillons 1,

CH-2800 Delemont, Switzerland, 3. Department of Biology, Colorado State University,

Fort Collins, CO 80523-1177, USA, 4. Department of Land Resources and Environmental

Sciences, Montana State University, Bozeman MT 59717-3120, USA, 5. Department of

Wildland Resources, Utah State University, Logan UT 84322-5230, USA, 6. Department of

Biology, Pennsylvania State University, University Park PA 16802-5301.

The success of suppressing densities of invasive plants by re-introducing natural

enemies from the weed’s native range may be quite variable across the invaded range. One

important source of variation may be the biocontrol agents themselves. Often they originate

from multiple locations, and it is usually unknown which introduction led to establishment or

whether agents from different locations have hybridized. Hybridization between genetically

distinct populations can be beneficial (e.g. hybrid vigour) or detrimental (e.g. outbreeding

depression) to biological control, and can drive rapid eco-evolutionary dynamics. We

genotyped 15 ragwort flea beetle ( Longitarsus jacobaeae) populations in northwestern

Montana, USA where beetles originating from Italy and from Switzerland were released to

control the noxious weed, tansy ragwort ( Jacobaea vulgaris). We found that almost half of

the populations contained hybrid individuals, while the rest were of Swiss origin and none

of pure Italian origin. We then tagged 60 tansy ragwort rosettes at each of 9 sites where

sufficient numbers of plants were available: 6 with Swiss beetles only and 3 containing

hybrids. To assess the efficacy of biocontrol we applied insecticide to half of the tagged

plants (the controls) to exclude beetles, and we monitored the plants for 2 years. Plants that

were exposed to ambient levels of feeding by the flea beetles had 52% higher mortality, 17%

lower flowering success, and those that flowered had 59% lower seed production than low

herb ivory plants. At hybrid sites plant mortality tended to be higher, and flowering success

JESO Volume 147, 2016

and seed production lower, although only plant fecundity differed significantly from sites

with Swiss beetles. Our results indicate that Swiss beetles provide successful control of

tansy ragwort at high elevation sites in Montana, where pure Italian beetles had difficulties

establishing due to a mismatch with the climate. Moreover, hybridization between Swiss

and Italian beetles may have benefited biocontrol, since hybrids appear more effective

than Swiss beetles. These findings reveal the power of biological control, and highlight the

importance of hybridization in shaping the outcome of biocontrol programs, and suggest

that taking an evolutionary perspective in implementing biocontrol could increase success

rates.

JESO Volume 147, 2016

THE ENTOMOLOGICAL SOCIETY OF ONTARIO

OFFICERS AND GOVERNORS

2016-2017

President: G. OTIS

School of Environmental Sciences,

University of Guelph, Guelph, ON NIG 2W1

gotis@uoguelph.ca

President-Elect: A. GUIDOTTI

Department of Natural History, Royal Ontario Museum

100 Queen’s Park, Toronto, ON M5S 2C6

antoniag@rom. on. ca

Past President: J. GIBSON

Royal BC Museum

675 Belleville Street, Victoria, BC V8W 9W2

JGibson@royalbcmuseum. be. ca

Secretary: M. LOCKE

Vista Centre, 1830 Bank St, PO Box 83025

Ottawa, ON K1V 1A3

entsocont. membership@gmail. com

Treasurer: S. LI

Natural Resources Canada, Canadian Forest Service

960 Carling Ave., Building 57

Ottawa, ON K1A 0C6

sli@nrcan.gc.ca

Directors:

D. BERESFORD (2015-2017)

Biology Department, Trent University

1600 West Bank Drive,

Peterborough, ON K9J 7B8

davidberesfbrd@trentu. ca

J. KITS (2017-2019)

Agriculture and Agri-Food Canada,

K. W. Neatby Building, 960 Carling Avenue,

Ottawa, ON K1A 0C6

Joel.Kits@agr.gc.ca

A. ROE (2017-2019)

CFS - GLFC | SCF - CFGL,

1219 Queen Street East

Sault Ste. Marie, ON

amanda.roe@canada. ca

A. SMITH (2016-2018)

Department of Integrative Biology,

University of Guelph, 50 Stone Road East

Guelph, ON NIG 2W1

salex@uoguelph. ca

J. SMITH (2015-2017)

University of Guelph Ridgetown Campus

120 Main St. E, Ridgetown, ON NOP 2C0

jocelyn. smith@uoguelph. ca

L. TIMMS (2016-2018)

laura. le. timms@gmail. com

ESO Regional Rep to ESC: S. CARDINAL

Agriculture and Agri-Food Canada,

K. W. Neatby Building, 960 Carling Ave,

Ottawa, ON K1A 0C6

sophie. cardinal @agr. gc. ca

Webmaster: T. BURT

trevburt@gmail .com

Newsletter Editors:

L. DES MARTEAUX

Department of Biology,

University of Western Ontario

Biological and Geological Sciences Building

London, ON N6A 5B7

ldesmart@uwo.ca

K. SHUKLA

University of Western Ontario

kruti. shukla@gmail .com

Student Representatives:

C. SCOTT

University of Toronto

Catherine. elizabeth. scott@gmail. com

A. YOUNG

Agriculture and Agri-Food Canada

K.W. Neatby Building, 960 Carling Avenue,

Ottawa, ON K1A 0C6

a.d.young@gmail.com

JESO Editor: C. MACQUARRIE

Natural Resources Canada, Canadian Forest Service

Great Lakes Forestry Centre

1219 Queen Street East,

Sault Ste. Marie, ON P6A2E5

chris. macquarrie@NRCan-RNCan. gc. ca

Technical Editor: T. ONUFERKO

Department of Biology, York University

Lumbers Building (RM 345), 4700 Keele Street

Toronto, ON M3J 1P3

onuferko@yorku.ca

ENTOMOLOGICAL SOCIETY OF ONTARIO

http: //www. entsocont. ca

The Society founded in 1863, is the second oldest Entomological Society in North America

and among the nine oldest, existing entomological societies in the world. It serves as an

association of persons interested in entomology and is dedicated to the furtherance of

the science by holding meetings and publication of the Journal of the Entomological

Society of Ontario. The Journal publishes fully refereed scientific papers, and has a

world-wide circulation. The Society headquarters are at the University of Guelph. The

Society’s library is housed in the McLaughlin Library of the University and is available

to all members.

An annual fee of $30 provides membership in the Society and a subscription to the

Newsletter. Students, amateurs and retired entomologists within Canada can join free

of charge. Publication in and access to the Journal is free to all members and non¬

members.

APPLICATION FOR MEMBERSHIP

Please send your name, address (including postal code) and email address to:

Michelle Locke, Secretary, Entomological Society of Ontario

c/o Vista Centre, 1830 Bank Street, P.O. Box 83025 Ottawa, ON K1V 1A3

or email: entsocont.membership@gmail.com

NOTICE TO CONTRIBUTORS

Please see the Society web site (http://www.entsocont.ca) for current instructions to

authors. Manuscripts can be submitted by email to the Scientific Editor

(JESOEditor@gmail. com).

FELLOWS OF THE ENTOMOLOGICAL SOCIETY OF ONTARIO

W. W. BILL JUDD

C. RON HARRIS

GLENN WIGGINS

2002 FREEMAN MCEWEN

2003 THELMA FINLAYSON

2006 JOHN STEELE

2010

2013

2014

BERNARD PHILOGENE

CONTENTS

BLANK.1-2

I. ARTICLES

J. SAVAGE — First Canadian record of the Bermuda grass stem maggot, Atherigona reversura

(Diptera: Muscidae).3-6

T. SHARMA, J. DEDES, and C. J. K. MACQUARRIE — A modified technique for rearing

wood boring insects permits visualization of larval development.7-14

J. M. RENKEMA, B. G. EVANS, C. HOUSE, and R. H. HALLETT — Exclusion Fencing

Inhibits Early-Season Beetle (Coleoptera) Activity-Density in Broccoli.15-28

II. POSTER AND PRESENTATION ABSTRACTS — ENTOMOLOGICAL SOCIETY

OF ONTARIO ANNUAL GENERAL MEETING.29-41

III. ESO OFFICERS AND GOVERNORS 2016-2017.42

IV. ESO OFFICERS AND GOVERNORS 2015-2016.inside front cover

V. FELLOWS OF THE ESO.inside back cover

VI. APPLICATION FOR MEMBERSHIP.inside back cover

VII. NOTICE TO CONTRIBUTORS.inside back cover

JOURNAL OF THE ENTOMOLOGICAL SOCIETY OF ONTARIO - VOLUME 147, 2016