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Species Composition, Monitoring, and Feeding Injury of StinkBugs (Heteroptera: Pentatomidae) in BlackberryAuthor(s): S. A. Brennan , O. E. Liburd , J. E. Eger , and E. M. RhodesSource: Journal of Economic Entomology, 106(2):912-923. 2013.Published By: Entomological Society of AmericaURL: http://www.bioone.org/doi/full/10.1603/EC12415

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HORTICULTURAL ENTOMOLOGY

Species Composition, Monitoring, and Feeding Injury of Stink Bugs(Heteroptera: Pentatomidae) in Blackberry

S. A. BRENNAN,1,2 O. E. LIBURD,1 J. E. EGER,3 AND E. M. RHODES1

J. Econ. Entomol. 106(2): 912Ð923 (2013); DOI: http://dx.doi.org/10.1603/EC12415

ABSTRACT Blackberry (Rubus spp.) production in Florida has increased �100% within the past twodecades. and several insect pests, including stink bugs (Heteroptera: Pentatomidae), have beenobserved feeding on this crop. The objectives for this study were to determine the stink bug speciespresent in blackberry; to develop monitoring tools for stink bugs in blackberry; and to describe feedinginjury to blackberries by Euschistus quadrator Rolston, a relatively new stink bug pest to Florida, thathas spread throughout the state. In a Þeld survey,E. quadratorwas the most abundant stink bug species,followed by Euschistus servus Say, Euschistus obscurus (Palisot de Beauvois), Thyanta custator (F.),Proxys punctulatus (Palisot de Beauvois), and Podisus maculiventris Say. Yellow pyramid traps caughtmore stink bugs than tube traps with or without the addition ofEuschistus spp. pheromone lures. Therewere no statistical differences between traps baited with a Trece Pherocon Centrum lure, a SuterraScenturion lure, and an unbaited trap. These results were supported by Y-tube olfactometer assayswith E. quadrator where there were no differences between pheromone baited lures and a control.Injury to berries caused by E. quadrator adults and third instars was similar, and both adults and thirdinstars fed more on green berries compared with turning berries. In addition, adults fed more on greenberries compared with ripe fruit. The most common injury to green berries was discoloration. Incontrast, misshapen drupelets were commonly seen on turning and ripe berries. The potential formanaging stink bugs in blackberries to prevent them from reaching damaging levels is discussed.

KEY WORDS blackberry, stink bug, Euschistus spp., injury, monitoring

Several blackberry (Rubus spp.) cultivars have re-cently been developed to be more tolerant to theclimatic conditions of the southern United Sates, pos-sibly opening this region to further commercial black-berry production (Jennings et al. 1991, Moore 1997).Georgia has tripled its production in the past 10 yr,with 122 ha of blackberries harvested in 2007 (Strik etal. 2007, USDAÐNASS 2009). Similarly, Florida black-berry production has increased Þvefold within thepast 5 yr (O.E.L., unpublished data). Earlier ripeningof blackberries in Florida may enable growers to meeta market window not currently being Þlled by othersouthern states.

Many pest species present in other geographicalareas also affect blackberries in the southeast. South-ern green stink bug, Nezara viridula (L.); green stinkbug, Acrosternum hilare (Say); and Euschistus spp.have been reported as pests of blackberries in thesoutheastern United States (Johnson and Lewis 2003,Anonymous 2008, Mizell 2008). The brown stink bug,Euschistus servus (Say), is also known to cause injuryto other Rubus spp. (Maxey 2011).

The genusEuschistusnot only containsE. servus butalso other pest species in the continental UnitedStates, including Euschistus variolarius (Palisot deBeauvois), Euschistus tristigmus (Say), Euschistus ic-tericus(L.),Euschistus quadratorRolston, andEuschis-tus conspersus (Uhler). E. quadrator and smaller spe-cies in the genus, such as Euschistus obscurus (Palisotde Beauvois), are sometimes referred to as the “lesserbrown stink bug complex” due to their increasing peststatus (Hopkins et al. 2005). Members of the genusEuschistus have been observed feeding on many cul-tivated crops, including corn, Zea mays L.; cotton,Gossypium hirsutum (L.); alfalfa, Medicago sativa L.;soybean, Glycine max (L.) Merr.; and various fruits(McPherson and McPherson 2000). These specieshavemigrated south, and increased inpest status in thesoutheastern United States in recent years. The intro-duction of crops such as Bt cotton in the south, whichdoes not control stink bugs, and reduced applicationof broad-spectrum insecticides in favor of more se-lective pesticides may have contributed to the in-crease in numbers of Euschistus spp. in southeasternplantings. In addition to stink bugs, thrips (Franklin-iella spp.); twospotted spider mites, Tetranychus urti-caeKoch; and gall midges (Dasineura spp.) have beenlisted as pests of concern for Florida (Ellis et al. 1991,Mizell 2007; O.E.L. et al., unpublished data).

1 Department of Entomology and Nematology, University of Flor-ida, Bldg. 970 Natural Area Dr., Gainesville, FL 32611.

2 Corresponding author, e-mail: sbrennan@uß.edu.3 Dow AgroSciences LLC, 2606 S. Dundee St., Tampa, FL 33629.

0022-0493/13/0912Ð0923$04.00/0 � 2013 Entomological Society of America

Stink bugs are highly mobile and polyphagous pests,thereby making monitoring difÞcult (McPherson andMcPherson 2000). Visual searches, ground cloths,blacklight traps, fruit injury, beating, and sweep-netsamples have typically been used to determine stinkbug infestation levels in various crops (Todd and Her-zog 1980). Many of these methods are labor-intensiveand can be biased against catching nymphs or adults(Toews and Shurley 2009).

Several species of stink bugs produce pheromonesthat aid in the aggregation of stink bugs in the Þeld(Todd and Herzog 1980, Krupke et al. 2001). It isbelieved that these male-produced pheromones at-tract both sexes (Aldrich et al. 1991, Millar et al. 2002,Leskey and Hogmire 2005) and may have potential inimproving monitoring efÞciency in the Þeld.

Aldrich et al. (1991) trapped volatiles separatelyfrom male and female Euschistus spp. by using gaschromatography and found a sex-speciÞc component,methyl (2E,4Z)-decadienoate, produced by males.This pheromone attracts males, females, and nymphsof severalEuschistus spp. and is a major component forE. servus, Euschistus politus (Uhler), E. ictericus, E.conspersus, and E. tristigmus (Aldrich et al. 1991).Although direct studies of volatile secretions of E.quadrator have not been conducted, this species iscaught in traps baited with the Euschistus spp. pher-omone (Tillman and Cottrell 2012; S.A.B., unpub-lished data).

Mizell and Tedders (1995) modiÞed a pyramid trapinitially developed by Tedders and Wood (1994) tomonitor for the pecan weevil,Curculio caryae (Horn).These pyramid traps are most effective when paintedwith industrial safety yellow as opposed to other col-ors, indicating that the color yellow may be an attrac-tive visual stimulus for stink bugs (Mizell and Tedders1995, Leskey and Hogmire 2005). Yellow pyramidtraps have increased attraction when paired with theEuschistus spp. pheromone methyl (2E,4Z)-decadi-enoate. Hogmire and Leskey (2006) were able to sig-niÞcantly reduce escape ofEuschistus spp. by reducingthe cone opening inside the trap, and increasing jarsize, with or without an insecticidal ear tag in apple(Malus spp.) and peach (Prunus spp.) orchards.

Tube traps made from clear plastic tubes with wiremesh cones on the ends have been used in severalstudies with varying results, and they are still com-mercially available for monitoring stink bugs. Krupkeet al. (2001) caught very few stink bugs by using twovariations of the tube trap in mullein (Verbascumthapsus L.), whereas Aldrich et al. (1991) used thesetraps to evaluate the Euschistus spp. pheromone inweedy areas or blackberry patches, and these trapscaught many stink bugs.

Very little research has been conducted on stinkbugs in blackberries, despite their potential to causefruit injury. Feeding injury in blackberries results incollapsed or leaky drupelets that render the fruit un-marketable. Moreover, stink bug odor can alter thetaste of the fruit and thereby also negatively affect itsmarketability. This is especially important in mechan-ically harvested berries where stink bugs are contin-

uously disturbed with the mechanical harvester (De-Francesco et al. 2002).

With the changing pest complex and potential fu-ture increases in blackberry production in the south-east, it is necessary to identify and survey any previ-ously undocumented pest species for blackberry inFlorida that may pose a threat to increasing produc-tion. The speciÞc objectives for this study were toidentify key stink bug species present in blackberry, tocompare commercial traps and lures for monitoringstink bugs in blackberries, and to conduct feedingassays to determine injury caused by E. quadrator onblackberry.

Materials and Methods

Study Site. Research was conducted at the Univer-sity of Florida, Plant Science Research and EducationUnit (PSREU) in Citra, FL, and at the Small Fruit andVegetable integrated pest management (SFVIPM)laboratory at the University of Florida in Gainesville,FL. The experimental site in Citra consists of two0.10-ha sites, each composed of six blackberry culti-vars, one cultivar per row: ÔKiowaÕ, ÔOuachitaÕ, ÔArap-ahoÕ, ÔChickasawÕ, ÔChoctawÕ, and ÔNatchezÕ. Each rowwas �38.1 m in length, with 30 blackberry plants. Rowswere spaced �4.5 m apart, with the plot �27.5 m inwidth. Half of the site was managed as a traditionalconventional site, and the other was managed as anorganic site. Blackberry plants were �3 yr old and1.5 m in height during experimentation.

Blackberry plots were watered using a hard linewith in-line emitters as needed, not exceeding 23,385Ð28,062 liters/ha/d. The organic plot received half theamount of water as the conventional plot. Organicplots were covered with DeWitt landscaping tarp(DeWitt Company, Sikeston, MO). Hydrosource wa-ter-gathering crystals (Castle International Re-sources, Sedona, AZ) in various sizes (medium andstandard) were added to the soil at a rate of 4,882.59kg/ha in the organic plots for water retention. Black-berries were grown on a moving arm shift trellis (Stiles1999). This trellis is shifted during bloom stages toallow ßowers to open toward the sky and after fruitingto ease in berry harvest. After fruiting, ßoricanes werepruned to allow room for the primocanes.

All fungicides were applied biweekly at manufac-turerÕs recommended rates. Both conventional andorganic plots were sprayed with copper sulfate(LoweÕs, Gainesville, FL) as a fungicide. Conventionalplots also received the fungicide Pristine (BASF, Co.,Research Triangle Park, NC), whereas organic plotswere sprayed with the fungicide Serenade (Agra-Quest, Inc., Davis, CA). With respect to nutrition, inMarch and June conventional plots were fertilizedwith 10Ð10Ð10 (LoweÕs) and the organic plots werefertilized with Nature Safe 10Ð2Ð8 (Nature Safe ColdSpring, KY) at a rate of 224.17 kg/ha over a 1-m band.Weed control in the conventional plots was accom-plished manually or with the herbicide Round Up(Monsanto, St. Louis, MO) at the recommended rate,

April 2013 BRENNAN ET AL.: STINK BUGS IN BLACKBERRY 913

whereas weeds were hand-pulled weekly from theorganic plot.Species Composition. Sampling was conducted in

2010 on four randomly chosen blackberry plants ofeach cultivar in conventional and organic blackberryplots. A harvest tray (�1 by 0.6 m) (Wal-Mart, Gaines-ville, FL) was placed on the ground at the base of theblackberry bush. Bushes were shaken vigorously threeto four times over the tray, and the cover of the traywas replaced immediately. All stink bug species thatfell into the tray were collected. Sampling occurredonce every 2 wk for 8 wk from 8 May to 19 June whenblooms were �50% fruiting. Stink bugs from eachsample were transferred to collecting jars and labeledby date, sample, variety, and plot. All jars werebrought back to the SFVIPM laboratory. All stink bugswere counted, pinned and identiÞed to species usinga key to the Florida families of Pentatomidae (J.E.E.,unpublished; Rolston 1974, McPherson 1989, Riderand Chapin 1992). Weekly observations on percent ofberries in various ripening stages were recorded.Trap Comparison. Two different types of commer-

cially available stink bug traps were compared withand without pheromone lures: 1) Yellow pyramid trap(R. Mizell, Quincy, FL) and 2) Trece tube trap (GreatLakes IPM, Vestaburg, MI). Pyramid traps were con-structed as recommended by Mizell (2008). Both theTrece Pherocon Centrum lure (Trece Inc., Adair, OK)and Suterra Scenturion lure (Suterra Corporate,Bend, OR) were used in each baited trap becauseinformation on the lure that performs the best inblackberry was unavailable.

Four treatments were evaluated: 1) Pyramid trapbaited, 2) Pyramid trap unbaited, 3) Trece trap baited,and 4) Trece trap unbaited. The experiment was arandomized complete block design with three repli-cates. The yellow pyramid trap was placed east of therow �0.5 m from the bushes (Fig. 1). The Trece tubetrap was hung from the trellis inside the bush �1 mfrom the ground (Fig. 2). Traps were spaced a mini-mum of 15 m apart and were blocked by cultivar(row). Traps were placed in the organic side of theplanting. Trap contents were emptied into collectingjars, and treatments were rotated weekly for 3 wk. Allstink bug species were brought back to the SFVIPMlaboratory to be counted, pinned, and identiÞed.Pheromone Comparison. Insect Source. A labora-

tory colony was established and maintained for severalmonths from wild-caught E. quadrator adults from avariety of host plants. Adults were caged in groups of8Ð10 males and females. Cages (15 by 15 by 18 cm)were made from plastic food storage containers (Tar-get, Minneapolis, MN) with holes in the lids (� 4 by5 cm) and sides covered with 0.3-mm mesh screens foraeration. Each cage contained a 59.2-ml Solo soufßecup with lid (Solo Cup Company, Lake Forest, IL)with a cotton roll (1 cm in diameter, cut to 5-cmlength; Richmond Dental, Charlotte, NC) insertedinto a hole cut into the lid for a water source. Adultswere reared on organic green beans, Solanum lyco-persicum var. cerasiforme, organic roma beans, Phaseo-lus vulgaris L., and organic cherry tomatoes, Phaseolus

vulgaris “Roma II” L. A strip of cheesecloth was taped tothe inside of the cage for oviposition. Cages were kept ina rearing incubator at 25 � 0.5�C, 55 � 5% RH, and aphotoperiod of 16:8 (L:D) h.Laboratory Study. Y-tube methodology was modi-

Þed from Borges and Aldrich (1994) and Borges et al.(2011). Bioassays were conducted in a glass Y-tubeolfactometer (Chemglass Life Sciences, Vineland,NJ). HumidiÞed airßow was maintained at 1,000 ml/min by using a two-channel air delivery system, withtwo glass ßowmeters, an acrylic chassis, two charcoalÞlters, and two gas bubblers (Analytical Research Sys-tems, Gainesville, FL). The glass Y-tube (150-mmmain tube, 80-mm arms, 35-mm internal diameter,40/35 joints) was held at a 30� angle inside a cardboardbox (44 by 30 by 23 cm) on a piece of foam core setat a 5% incline (Fig. 3). Preliminary studies indicatedthat E. quadrator responded better at a 5% incline asopposed to a horizontal surface. The foam core andinterior of the box were white. A hole was cut into theside of the box at the end of the Y-tube so that pher-omones did not accumulate inside the box. Two masonjars, located outside of the box, were modiÞed tohouse the lures during the assays (Fig. 3). Holes weredrilled into the lids, and valves were secured to the lidsthat were connected to the corrugated plastic tubing(5.55-mm interior diameter and 6.35-mm outer diam-eter) (Cole Parmer, Vernon Hills, IL) attached to theÞltration system.

Two commercially available aggregation phero-mone lures,TrecePheroconCentrumlureandSuterra

Fig. 1. Yellow pyramid trap with screen top. (OnlineÞgure in color.)

914 JOURNAL OF ECONOMIC ENTOMOLOGY Vol. 106, no. 2

Scenturion lure, for monitoring Euschistus spp. werecompared in the Y-tube bioassay. Treatments werecompared as follows: 1) Trece Pherocon Centrum lureagainst a blank control, 2) Suterra Scenturion lureagainst a blank control, and 3) Trece Pherocon Cen-trum lure versus Suterra Scenturion lure. Stink bugswere placed in the Y-tube base and allowed to accli-mate for 3 min before attaching the Y-tube arms andairßow. Stink bugs were evaluated to determine theirpreference, and the time it took them to make a de-cision was recorded. A choice was considered madeafter the insect remained in one of the arms for 1 min.Stink bugs were considered unresponsive after stayingin the Y-tube for 15 min without making a choice.Adult stink bugs �1 mo old, from the colony, wereused in the assay. In total, 20 responding stink bugs pertreatment (10 males and 10 females) were evaluated,with the jars being rotated after 10 responding stinkbugs to prevent positional bias. Glassware and tubingwere rinsed with deionized water, methanol, and eth-anol and dried in an oven at 160�C after each test.Field Study. The aggregation pheromone lures

tested in the laboratory were evaluated in the Þeld.Yellow pyramid traps were used because our prelim-

inary trap comparison research indicated that thesetraps performed better than tube traps. Three treat-ments were compared: 1) Trece Pherocon Centrumlure, 2) Suterra Scenturion lure, and 3) unbaited trap.Experimental design was a randomized completeblock with four replicates. Traps were placed east ofthe row �0.5 m from the blackberry bushes. Trapswere spaced a minimum of 15 m apart within rows, �5m between rows, and were blocked by cultivar (row).Traps were placed in the Þrst, third, Þfth, and sixthrows of the organic side of the planting. Trap contentswere emptied into collecting jars, and traps were ro-tated weekly. All stink bug species were brought backto the SFVIPM laboratory to be counted, pinned, andidentiÞed.Feeding Injury.No-choice experiments to evaluateE. quadrator feeding injury on blackberries were con-ducted in the SFVIPM laboratory. Experimental lay-out consisted of three treatments in a completely ran-domized design with four replicates. Treatmentsincluded 1) green fruit, 2) turning fruit (red stage),and 3) ripe fruit (black berries). Blackberry shootswith berries used in assays were harvested from theblackberry planting located at PSREU. Bushes were

Fig. 2. Tube trap. (Online Þgure in color.)

Fig. 3. Y-tube olfactometer set up. (A) Jar 1. (B) Jar 2. (C) Air Þltration system. (D) ModiÞed cardboard box. (E) Aerationhole. (F) Foam board.

April 2013 BRENNAN ET AL.: STINK BUGS IN BLACKBERRY 915

managed as discussed under Study Site. Blackberryshoots containing at least two berries were collectedfrom Kiowa blackberries. After shoots with berrieswere cut from blackberry canes, they were imme-diately placed into a foam cooler with ice packs tomaintain freshness until they were placed in bioas-say chambers. Each shoot was surrounded with afoam stopper and placed into a 50-ml vial, contain-ing 35 ml of tap water before being placed into abioassay chamber. Bioassay chambers consisted of32-oz plastic containers with lids that had a meshcovered hole.E. quadrator used for fruit injury assays were ob-

tained from the colony discussed under Insect Source.The experiment was conducted with E. quadratoradults and with third instars. Adults and nymphs usedin the experiment ranged between 20 (nymphs) and50 (adults) days old. Before conducting assays, allstink bugs were starved for a 24-h period. Three stinkbugs were released into each container. In experi-ments with adult E. quadrator, females were releasedinto half of the containers, and males were releasedinto the other half. The adult experiment was con-ducted in two trials of four replicates to produce a totalof eight replicates, four with females and four withmales. Stink bugs were allowed to feed for 72 h. Con-tainers were checked daily and the location of eachstink bug (berry, leaf, or other [e.g., container, stem]),was recorded. After 72 h, injury descriptions weremade by examining each berry for visible signs ofinjury and recording them.

After the injury assessment, berries and leaves wereremoved from stems and stored in small plastic con-tainers in the freezer. A double stain solution, a com-bination of Acid Fuchsin and Lignin Pink, (BioQuipProducts, Rancho Dominguez, CA) was used to stainstink bug stylet sheaths on the leaves and berries(Bowling 1979). The solution was diluted using theratio 1 ml of stain:10 of ml water. Berries and leaveswere left in the stain for 1 h and then rinsed with tap

water. Leaves and berries were examined under adissecting microscope. Leaves were examined only forthe presence of stylet sheaths to determine whetherE.quadrator feeds on leaves as well as fruit. In contrast,the number of stylet sheaths on each berry wascounted, and these counts were used to calculate theaverage number of sheaths per berry.Data Analysis. Species survey information is pre-

sented as total counts from organic and conven-tional due to low numbers of stink bugs found in theÞeld. Data on trap comparison and pheromone com-parison were analyzed using analysis of variance(ANOVA), and differences among means were de-termined using the least signiÞcant difference(LSD) mean separation test (0.05) (PROC GLM,SAS Institute 2008).

The Y-tube assay data were analyzed using a chi-square analysis with an expected probability of 0.5. At-test was used to determine any signiÞcant differ-ences between sexes of Euschistus spp. (0.05) (PROCT-TEST, SAS Institute 2008).

For the feeding injury study, the number of E.quadrator found on berries was square rootÐtrans-formed to Þt the assumptions of the analysis and com-pared among berry stage (green, turning fruit, andripe fruit) and sex by using a two-way ANOVA (SASInstitute 2008) for the adult data. A one-way ANOVA(SAS Institute 2008) was used to compare the numberof E. quadrator found on berries for the third-instardata. Means were separated using the LSD test (SASInstitute 2008).

For the adult stink bug data, numbers of styletsheaths per berry were log10-transformed to Þt theassumptions of the analysis and compared using atwo-way ANOVA (SAS Institute 2008), with berrystage and sex as factors. For the third instars, numbersof stylet sheaths per berry also were log10-transformedto Þt the assumptions of the analysis and were com-pared using a one-way ANOVA (SAS Institute 2008).

Fig. 4. Species complex in blackberries by using the beating tray method. (Online Þgure in color.)

916 JOURNAL OF ECONOMIC ENTOMOLOGY Vol. 106, no. 2

Means were separated using the LSD test (SAS Insti-tute 2008).

Results

Species Composition. In total, 54 stink bugs werecollected in both conventional and organic blackberryplots throughout the sampling period. Figure 4 pro-vides the percentages of species collected, includingE. quadrator, the most abundant species, followed byE. servus; E. obscurus; Thyanta custator (F.); Proxyspunctulatus (Palisot de Beauvois); and the spined sol-dier bug, Podisus maculiventris (Say). Both males andfemales were found in the Þeld, with the exception ofP. punctulatus and P. maculiventris, for which no fe-males were recorded. Sex ratios between species werenot statistically different, averaging 1:1 (male:female)among most species. Also, there was no difference inspecies composition between conventional and or-

ganic plots. As the percentage of ripe fruit increasedin both the organic and conventional plots, the num-ber of stink bugs also increased (Fig. 5).

Other insects collected in the beating tray in-cluded ants (Hymenoptera: Formicidae), spiders(Arachnida: Araneae: Salticidae, Oxyopidae), grass-hoppers (Orthoptera: Acrididae), plant bugs(Hemiptera: Miridae), katydids (Orthoptera: Tet-tigoniidae), leaffooted bugs (Hemiptera: Co-reidae), lady beetles (Coleoptera: Coccinellidae),and ßower beetles [Coleoptera: Scarabaeidae; Eu-phoria sepulcralis (F.)]. There was no difference ininsect species composition between conventionaland organic plots.Trap Comparison. In Þeld tests, there was a highly

signiÞcant difference between trap types (F � 17.79;df � 3, 11;P� � 0.0001), with the yellow pyramid trapcatching more stink bugs than the tube trap (Fig. 6).However, there were no signiÞcant differences be-

Fig. 5. Total stink bug numbers correlated with mean percent fruit development in organic and conventional blackberryplots.

April 2013 BRENNAN ET AL.: STINK BUGS IN BLACKBERRY 917

tweenbaitedandunbaitedpyramidor tube traps.Bothmales and females were caught in the traps, but therewere no statistical differences between sexes (t� 0.49,P � 0.6526).

The species recorded for this experiment, in orderof abundance, consisted of E. servus, E. quadrator, P.maculiventris, E. obscurus, Euschistus ictericus (L.),and T. custator (Table 1).Pheromone Comparison. In the Y-tube assays,

there were no statistical differences between any ofthe treatments. In the Trece lure versus a blank con-trol, 12 stink bugs chose the lure and eight chose theblank control (�2 � 0.8, df � 1, P � 0.3711). In theSuterra lure versus a blank control, 10 stink bugs eachchose the lure or the blank control (�2 � 0.0, df � 1,P� 1.00). In the Trece lure versus the Suterra lure, 13stink bugs chose the Trece lure and seven chose theSuterra lure (�2 � 1.8, df � 1, P� 0.1797). There were12 nonresponders in total, distributed fairly evenlyacross all treatments.

Similar to our Y-tube assays, no signiÞcant differ-ences were found in the Þeld tests that comparedpheromone-baited pyramid traps with unbaited pyr-amid traps (F� 0.85; df � 2, 11; P� 0.4354) (Fig. 6).Numerically, traps baited with the Trece lure caughtmore stink bugs (n� 10). The species composition forthis experiment in order of abundance consisted of E.servus, T. custator, E. quadrator, and E. ictericus. Bothmales and females were caught in the traps, but therewere no statistical differences between sexes (t� 0.18,P � 0.8648).Feeding Injury. Injury to berries caused by E. qua-

drator adults and third instars was similar. The mostcommon injury to green berries was discoloration(Fig. 7A). There were a few green berries with mis-shapen drupelets (Fig. 7B). In contrast, misshapendrupelets were commonly seen on turning and ripeberries (Fig. 7B), with only a few berries showingdiscoloration (Fig. 7C).

In the comparison of numbers of E. quadrator onberries, there was no interaction between berry stageand sex on any sampling day in the adult data (all F�1.54; df � 2, 23; P� 0.24). There were no differencesbetween males and females on any sampling day ineither adult experiment (all F � 1.39; df � 1, 23; P �0.25). There were no differences among treatments(Fig. 8A) on any sampling day (all F� 1.24; df � 2, 23;P � 0.31).

In the experiment with third instars, 72 h afterrelease, there were signiÞcantly higher numbers ofnymphs on the ripe berries compared with the greenberries (F� 4.5; df � 2, 11; P� 0.04) (Fig. 8B). Therewere no differences on any other day (both F� 1.46;df � 2, 11; P � 0.28).

In comparing the number of stylet sheaths perberry, there was no interaction between berry stageand sex in the adult (F � 0.27; df � 2, 23; P � 0.77).There was also no difference between males and fe-males (F � 2.81; df � 2, 23; P � 0.11). For the adults,there were signiÞcantly more stylet sheaths found ingreen berries compared with both of the other berrystages (F � 16.04; df � 2, 23; P � 0.0001) (Fig. 9).

Fig. 6. Mean number of stink bugs captured in yellow pyramid traps (YP) or tube traps with (B) or without (UB)pheromone lures. Means followed by the same letter were not signiÞcantly different at P � 0.05 level.

Table 1. Mean species composition for trap comparison study(n � 35)

Species Avg. densitya % totalb

E. servus 5.00 � 3.05 42.86Males 1.67 � 0.67Females 3.33 � 2.40E. quadrator 2.00 � 0.00 17.14

Males 1.67 � 0.33Females 0.33 � 0.33T. custator 0.33 � 0.33 2.86

Males 0.33 � 0.33Females 0.00E. ictericus 1.33 � 0.88 11.43

Males 0.33 � 0.33Females 1.00 � 0.58P. maculiventris 1.67 � 0.67 14.29

Males 0.00Females 1.67 � 0.67E. obscurus 1.33 � 0.33 11.43

Males 0.33 � 0.33Females 1.00 � 0.58

a Average density (mean � SE) of each category per replicate.b Percentage of total stink bugs collected.

918 JOURNAL OF ECONOMIC ENTOMOLOGY Vol. 106, no. 2

In the experiment with third instars, there weresigniÞcantly more stylet sheaths found in green ber-ries compared with turning berries (F � 5.34; df �2, 11; P � 0.03).

Stylet sheaths were found in leaves from all berrystages in all three experiments. Sheaths were found inleaf veins and were concentrated on the main leafvein.

Discussion

Several new stink bug records for blackberries werefound. This report is the Þrst known record of E.quadrator, E. obscurus, T. custator, P. punctulatus, andP. maculiventris in blackberry. Little, if any, informa-tion has been published in journals on the stink bugcomplex in blackberries. The limited informationavailable is on websites states that green stink bugs,southern green stink bugs, and brown stink bugs aremost prominent.

The Þrst visual sighting of stink bugs was ofE. servuson 28 April 2010 in the organic plot that began fruitingafter the conventional plot. Blackberries ripenedfaster in the organic plot, perhaps making the organicplot more attractive to stink bugs. Although overall

stink bug numbers were low, there was a generalcorrelation between the number of stink bugs foundon each sampling date and the amount of ripe fruit ineach plot. As the percentage of ripe fruit increased inboth the organic and conventional plots, the numberof stink bugs also increased. During sampling, moststink bugs were found on the third sampling date (5June 2010), when most berries were ripe. The Chick-asaw cultivar produced the most stink bugs across boththe conventional and organic plots. This variety ripens�1 wk later than Kiowa and Natchez cultivars.

The majority of stink bug information in Florida,especially for Euschistus spp., is from the early 20thcentury. A review of the literature shows that, histor-ically, the pest status of stink bugs in Florida is debat-able. The most commonly mentioned stink bugs in thestate are N. viridula, E. servus, and E. ictericus. N.viridula is commonly found in rice (Oryza sativa L.),soybean, faba bean (Vicia faba L.), and various weedsin southern Florida (Buschman and Whitcomb 1980,Temerak and Whitcomb 1984, Jones and Cherry 1986,Nuessly et al. 2004, Cherry and Wilson 2011). Giventhis information, we expected to Þnd mostly greenstink bugs and E. servus in our blackberry planting. E.servus is usually seen in pecan, Carya spp., soybean,

Fig. 7. Stink bug injury to berries. (A) Discoloration of green berry. (B) Ripe berry with misshapen drupelet. (C)Discoloration of ripe berry. (Online Þgure in color.)

April 2013 BRENNAN ET AL.: STINK BUGS IN BLACKBERRY 919

elderberry, Sambucus spp., and goldenrod Solidagospp., in Florida (Hill 1938, Frost 1979, Fontes et al.1994). E. ictericus is found in rice and faba bean (Te-merak and Whitcomb 1984, Jones and Cherry 1986,Nuessly et al. 2004, Cherry and Wilson 2011). There isno doubt that these pests also occur in other Floridacrops, but literature with this information was notfound.

When using any monitoring device, care should betaken in identifying the stink bug species present. E.servus is relatively easy to identify versus other Eu-schistus spp., butE. ictericus, E. tristigmus, andP.macu-liventris look very similar, and the smaller brown stinkbugs of the “lesser brown stink bug complex” are easilyconfused (Hopkins et al. 2005). Predatory stink bugs,such as the spined soldier bug, may be mistaken aspests. P. maculiventris is a beneÞcial predatory stinkbug thatmostly feedson lepidopteranandcoleopteranlarvae, but it has been shown to feed on phytophagousstink bugs (McPherson et al. 1980, McPherson andMcPherson 2000).

The yellow pyramid traps captured both predatoryand pest species by using the Euschistus spp. phero-mone (Mizell 2008). In the trap comparison experi-ment, the yellow pyramid trap was clearly superior. Infact, no stink bugs were caught in the tube traps. Thisis similar to the observations of Krupke et al. (2001),where very low numbers of stink bugs were capturedwhen comparing two different sizes of tube traps.However, it should be noted that Aldrich et al.(1991) caught a number of Euschistus spp. by usingthe tube trap in a deciduous forest. This indicatesthat tube traps may not be effective in blackberryplantings but may have potential uses in other crops.The pyramid traps were effective in catching stinkbugs, either with or without the addition of theEuschistus spp. pheromone, and there were no dif-ferences between baited and unbaited traps. Moststudies using the pyramid trap Þnd that the additionof the Euschistus spp. pheromone increases the ef-Þcacy of the trap (Leskey and Hogmire 2005, Cot-trell and Horton 2011). It is possible that volatiles

Fig. 8. Number of stink bug adults (A) and third instars (B) on berries at each sampling time. Means with the same letterare not signiÞcantly different from each other at P � 0.05. Error bars represent SEM.

Fig. 9. Average sheaths per berry recorded from each berry stage from the Þrst and second adult and third-instar larvaeexperiments. Means with the same letter are not signiÞcantly different from each other at P� 0.05. Error bars represent SEM.

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emitted from developing blackberries may havelessened the effect of the pheromone, making it lessattractive to Euschistus spp.

Similarly, in the pheromone comparison experiment,there were no statistical differences between the TrecePherocon Centrum lure, the Suterra Scenturion lure,and an unbaited trap. Overall, stink bug numbers werelow, which may partially account for some of the ob-served nonsigniÞcant differences and the obvious simi-larities ineffectiveness.However, theseresultsweresup-ported by the Y-tube olfactometer assays where therewere no statistical differences between the lures and acontrol, although higher numerical values were re-cordedwith theTrece lurewhenusingE.quadrator.Thecomposition of these commercial lures was unknownbecause this was proprietary information that the com-panies did not want to disclose. Because there were nostatisticaldifferencesbetweenlures,both intheÞeldandin the Y-tube bioassay, there is strong evidence to indi-cate that these lures may not be effective for monitoringstink bugs in blackberry plantings. Also, that there wereno differences between baited and unbaited traps mayindicate that the primary attraction to pyramid traps inblackberry crops may be a response to the yellow colorand probably trap type (architecture).

Species found in Þeld experiments included E. qua-drator, E. servus, E. obscurus, T. custator, E. ictericus, andP. maculiventris. Only adult stink bugs were caught inmonitoring traps. The reason why only adults werecaught is most likely due to adult stink bugs ßying to thetraps, and nymphs were not likely to crawl to the trapsfrom blackberry bushes. Because no statistical differ-ences were found between sexes, it can be determinedthat this trapattractsbothmalesandfemales.LeskeyandHogmire (2005) captured both sexes of E. servus in asimilar pyramid trap; however they found a higher per-centage of females than males in both baited and un-baited traps. Stink bugs seem to colonize blackberrywhen berries are mid-ripe to fully ripe. In the trap com-parison Þeld experiment the majority of stink bugs werefound in traps located in theKiowavariety.Kiowaripensearlier than most varieties, in early June, and fruitingextends for 6 wk (Moore and Clark 1996). The peak instink bug numbers occurred during the week of 5 May2010, when berries were mid-ripe. In the pheromonecomparison experiment, most stink bugs were foundduring the week of 26 May 2010 in Kiowa and Natchezwhen berries were beginning to ripen. Natchez alsoripens early in June, and fruiting extends for 4 wk (Clarkand Moore 2008).P. punctulatus was not caught in traps during Þeld

studies but was found in the species survey experi-ment. This stink bug has been found previously in thepyramid trap (Mizell 2008). It has been suspected ofbeing predaceous but is also known to feed on plantsand weeds, such as cotton and zigzag spiderwort, Tra-descantia subaspera Ker (Vangeison and McPherson1975, Gomez and Mizell 2009). These stink bugs arenot thought to cause signiÞcant economic damage(Schaefer and Panizzi 2000).E. quadrator feeding injury to green blackberries

produced discoloration but little deformation. This

was probably due, in part, to berries not continuing todevelop after the shoots are clipped from the bushes.Deformation injuries to turning and ripe berries weremore obvious.

Berry stage had little effect on the location of E. qua-drator within the containers. This may be partly due toboth adults and third instars feeding on the leaves as wellas the berries. Most of the adults and all of the nymphswere found on some part of the blackberry shoot.

The mean number of stylet sheaths found in thedifferent berry stages indicates that both adult andthird-instar E. quadrator fed more on green berriescompared with turning fruit and adults fed more ongreen berries compared with ripe fruit. Therefore,monitoring forE.quadrator should begin as soon as theblackberry plants begin to set fruit. Adults and nymphsalso fed on both turning and ripe berries. In the Þrsttrial of the adult experiment and the third-instar ex-periment, the turning fruit was in a later stage ofripening compared with the turning fruit in the secondtrial of the adult experiment. This may explain the lackof differences between turning and ripe fruit.

Overall, blackberries in Florida seem to have a differ-ent stink bug complex than that of other productionareas. Although we found severalEuschistus spp. that arecommonly mentioned in other areas, we did not Þndeither the green stink bug or the southern green stinkbug. More extensive sampling may be needed to captureadditional species. It is also interesting to note that somany differentEuschistus spp. were found on the black-berries at the same time and that E. quadrator was thedominant species. Our Þndings also indicate that theyellow pyramid trap was more effective than the tubetrap for monitoring stink bugs in blackberry. Further-more, our Þndings indicate that the addition of phero-mone lures to pyramid traps may not increase their at-tractiveness to stink bugs in blackberry. Both adults andnymphs of E. quadrator feed on green fruit, and thespecies seem to favor this stage of ripening. Therefore,werecommendtheimplementationofpestmanagementstrategies, including regular monitoring, when fruit isyoung and stink bug nymphs begin to appear in theplanting.

Acknowledgments

We thank employees of the Plant Science Research andEducation Unit in Citra, FL, for assisting with plot mainte-nance. Special thanks to the SFVIPM laboratory at the Uni-versity of Florida for Þeld assistance throughout the durationof the project. Special thanks to Nicole Benda for assistancewith the Y-tube bioassay. We thank Eileen Buss for revisionson an earlier draft of this article. This project was funded bya sustainable SHARE grant from the University of Florida.

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Received 1 October 2012; accepted 21 January 2013.

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