J. Appl. Entomol. Responses of stored-product Anoiide to pheromone lures nd plnt-derived voltiles R. M. Mhroof 1 & T. W. Phillips 2 1 Deprtment of Biologicl nd Physicl Sciences, South Crolin Stte University, Orngeurg, USA 2 Deprtment of Entomology, Knss Stte University, Mnhttn, USA Keywords Lsioderm serricorne, Stegoium pniceum, Anoiide, ttrctnt, Coleopter, host odour, trpping Correspondence Rizn M. Mhroof (corresponding uthor), Deprtment of Biologicl nd Physicl Sciences, South Crolin Stte University, 3 College Street NE, Orngeurg, SC 29117, USA. E-mil: rmhroof@scsu.edu Received: June 25, 27; ccepted: Octoer 24, 27. doi: 1.1111/j.1439-418.27.1251.x Astrct Experiments were conducted in different loctions to investigte responses of dult Stegoium pniceum nd Lsioderm serricorne (Col., Anoiide) to different commercilly ville or prototype frictions of their femle-produced sex pheromones. The results showed tht the numer of S. pniceum cptured in trps ited with the commercilly ville lures ws significntly higher thn those cptured in trps ited with the prototype stegoinone lures. The three commercilly ville rnds of serricornin lures investigted were eqully effective in cpturing L. serricorne. In relted study, we conducted 9-week trpping experiment to determine if responses of L. serricorne to serricornin cn e enhnced y the presence of host plnt odours. Trps were ited with serricornin lone, serricornin plus dried red chilli (Cpsicum frutescens L.) or red chilli lone. The results showed tht the numer of eetles cptured in trps ited with comintion of serricornin nd chilli voltiles were significntly higher thn in trps ited with pheromone or chilli voltiles lone, indicting tht potentil exist for improved monitoring or mss trpping of L. serricorne y comining pheromone with plnt-derived voltiles present in Cpsicum spp. Introduction The drugstore eetle, Stegoium pniceum (L.) (Col., Anoiide) nd the cigrette eetle, Lsioderm serricorne (F.) (Col., Anoiide), re mjor pests on wide vriety of dry nd durle stored griculturl products (Howe 1957; Lefkovitch 1967; Bry et l. 1974; Brr nd Chhl 198; Kohno et l. 1983; Jco nd Ushkumry 1991). These two storedproduct Anoiide re found throughout the tropicl nd sutropicl prts of the world, nd thrive well in wrm uildings in temperte countries (Powell 1931; Howe 1957; Lefkovitch 1967). Chllenges in utilizing currently ville mngement prctices for these insect pests, such s resistnce development to methoprene (Benezet nd Helms 1994) nd phosphine (Zettler nd Keever 1994) in L. serricorne, wrrnt more effective pest mngement pproches. The foundtion to successful integrted pest mngement progrmme is n effective monitoring system tht provides informtion on species nd density of insects, foci of infesttion, routes of insect entry nd chnges in pest popultion over time (Burkholder 199; Mueller et l. 199; Phillips 26). Synthetic pheromones re commonly used s lures in trps to monitor insect ctivities, to detect the presence of infesttions, to estimte popultion densities nd to determine tretment efficcy. The sex pheromone of S. pniceum, stegoinone [2,3-dihydro-2,3,5-trimethyl-6-(1-methyl-2-oxoutyl)-4Hpyrn-4-one] (Kuwhr et l. 1975) nd the sex pheromones of L. serricorne, nhydroserricornin (2,6-diethyl-3,5-dimethyl-3,4-dihydro-2H-pyrn) nd serricornin (4,6-dimethyl-7-hydroxynonn-3- one) (Burkholder 197) re produced y dult femles nd responded to y dult mles. Synthetic L. serricorne pheromone lures re widely used for detecting hidden infesttions in dry food-processing fcilities, wrehouses, retil nd grocery stores, nd other dry storges (Fustini 1985; Mueller et l. J. Appl. Entomol. 132 (28) 161 167 ª 28 The Authors Journl compiltion ª 28 Blckwell Verlg, Berlin 161
Evlution of ttrctnts for stored-product Anoiide R. M. Mhroof nd T. W. Phillips 199; Phillips et l. 2; Cmpell et l. 22; Ppdopolou nd Buchelos 22; Arogst et l. 23; Nnsen et l. 24). Unlike synthetic L. serricorne pheromone lures, commercil frictions of S. pniceum re less ville ecuse of chllenges posed y the lck of n effective or high-qulity procedure for synthesizing stegoinone. Host voltiles cn ply mjor role in ffecting ehviour of oth mle nd femle insects s direct ttrctnts, oviposition stimulnts or s potentil synergists to sex nd ggregtion pheromones. The phenomenon of comining host-relted voltiles nd synthetic pheromones to improve potency of ttrctnts used in trps hve een well documented in severl rk nd sp eetles (Lin et l. 1992; Lndolt nd Phillips 1997), ut studies on the synergistic effects of pheromones nd host-relted voltiles in stored-product insects hve not received much ttention (Chmers 199). Plnt-derived voltiles hve een used in limited occsions, s it to monitor stored-product insect popultion densities (Phillips 1997). A recent report y Mhroof nd Phillips (27) showed tht dult L. serricorne were strongly ttrcted to voltiles from dried chilli pepper (Cpsicum frutescens L.). We re, however, unwre of ny pulished report on the synergistic effect of comining plnt-derived voltiles with serricornin on trpping L. serricorne. Our gols in this study were to determine the reltive efficcy of commercilly ville or prototype frictions of pheromone lures for S. pniceum nd L. serricorne, nd to investigte if the efficcy of L. serricorne lures cn e enhnced when comined with host-derived voltiles. The rnds of commercil S. pniceum nd L. serricorne lures used in the study were selected sed on their vilility nd cost-effectiveness. The prototype lures were used in the study in order to evlute the optiml dosge of stegoinone or serricornin required to develop n efficient lure. Mterils nd Methods Trpping S. pniceum using different commercilly fricted stegoinone The study ws conducted in two retil pet supply stores for eight continuous weeks from Novemer to Decemer 25. Most of the products sold in these stores were feed for cts, dogs, smll nimls nd irds, nd these stores were infested with different stored-product insect pests, including S. pniceum, efore the study ws conducted. The two retil pet 162 supply stores elonged to the sme retil chin, were similr in product lyout, ut differed in floor dimension. In pet store 1, in ddition to the min store floor re (1263.5 m 2 with ceiling height of 1.7 m), storge re (995. m 2 with ceiling height of 1.7 m) ws present to the south of the uilding. Two entrnce doors were locted to the west nd north of the uilding nd two windows were present to the north nd west of the min store re. To the south of the storge re ws loding dock grge door. Pet store 2 (1467.9 m 2 with ceiling height of 1.7 m) hd two seprte storge res (83.6 m 2 nd 557.4 m 2 ), with min door to the south nd the dock door to the est nd no windows. Flight ctivity of S. pniceum ws monitored using four Allure dimond sticky trps TM (Whitmire Micro-Gen Ls, St Louis, MO). An Allure trp TM hs dimond-shped openings t oth ends, hs sticky interior surfce (14 cm 2 ) with dimension of 15. 18. cm. Trps were ited with the pheromone lures CSP1-type1, CSP1-type2, CSP1-type3 nd CSP2, which re different frictions of stegoinone, the femle-produced sex pheromone of S. pniceum. Type1, type2 nd type3 versions of CSP1 represent low, medium nd high pheromone concentrtions respectively. The relese formultions of the code-numered lures re provided in tle 1. Tle 1 Vrious synthetic sex pheromones for stored-product Anoiide showing their relese methods Insect species Stegoium pniceum Lsioderm serricorne Lure designtion Relese methods Comments CSP1 CSP2 CLS 1 CLS 2 CLS 3 CLS 4 CLS 5 Ruer sept with sored pheromone Product under development Pheromone reservoir Commercilly relesing through ville permele memrne Ruer septum with Commercilly sored pheromone ville Polyethylene vil Commercilly relesing through ville permele wll Pheromone reservoir Commercilly relesing through ville permele memrne Ruer sept with Lortory mde, sored pheromone technicl grde serricornin Ruer sept with sored pheromone Product under development J. Appl. Entomol. 132 (28) 161 167ª 28 The Authors Journl compiltion ª 28 Blckwell Verlg, Berlin
R. M. Mhroof nd T. W. Phillips Evlution of ttrctnts for stored-product Anoiide Pheromone lures used in ll experiments were within the ctive production period nd hd not pssed shelf-life. Ech trpping site ws divided into four locks. Five trps were rndomly deployed in ech lock, of which four of the trps were ited with pheromone lures nd the fifth ws without lure (control), giving totl of 2 trps per store. The trps were suspended verticlly on string, 1.8 m ove the ground level, out 2 m etween the trps, nd 3. m from the side wlls, nd plced in such wy s to minimize interference from customers nd store personnel. Trps were checked every 2 weeks during which the numer of S. pniceum cptured were recorded. Pheromone lures nd trps were replced every 4 weeks. A preliminry sttisticl nlysis ws conducted using nov to test for differences in distriutions of the trpping dt etween the two retil pet supply stores. The result showed tht the numer of eetles cptured did not differ significntly etween the sites. Thus, dt from the two sites were pooled in susequent sttisticl nlyses. Trpping L. serricorne using different commercilly fricted serricornin Two trpping experiments were crried out. The first ws conducted from August to Octoer 25 in multipurpose room t the Stored Products Reserch nd Eduction Center (SPREC) (36 7 N; 97 8 W) t Oklhom Stte University. The room mesured 8.5 m in length, 13.5 m in width nd height of out 6.5 m. Trpping ws crried out using the StorgrdÒ II sticky trps (Trécé, Adir, OK), designed to cpture flying insects with 14. 1.5 cm dhesive pnel nd dimondshped openings in either side. The room ws divided into five locks, with minimum of 8 m etween locks. Four trps were rndomly positioned in ech lock, of which three trps were ited with different commercil pheromone frictions herey designted s CLS1, CLS2 nd CLS3 (tle 1). The fourth trp ws left lnk (control), giving totl of 2 trps per week. The trps were suspended verticlly on string, 1.7 m ove the ground level, t distnce of 2. m from the wll, nd 1.5 m etween ech trp. A second L. serricorne trpping experiment ws conducted in the Animl Science Building (ASB) (36 7 N; 97 5 W) t Oklhom stte University from August to Octoer 25. The trpping ws conducted in the sement of two-storey uilding, mesuring 21.6 m long, 16.5 m wide nd 3.3 m tll. Within this spce n inner room (9.8 m 2 ) ws locted which ws seprted y two side wlls nd door. Trpping ws conducted using Storgrd ThinlineÒ (Trécé), rectngulr-shped sticky trp designed for trpping L. serricorne. The trp mesures 17.5 16 1.5 cm with n dhesive pnel of 15 cm 2. The trp is suitle for hnging on ropes or plcing underneth shelves, nd is opened on three sides of the rectngle for insect entry. To evlute the responses of L. serricorne to three commercilly fricted serricornin lures (CLS1, CLS3 nd CLS4) nd the fourth lure under development y the mnufcturer (CLS5; tle 1), rndomized complete lock design ws used with five locks (replictes). Lure type CLS5 hd three different concentrtions tht were lelled s type1, type2 nd type3 representing low, medium nd high pheromone concentrtions respectively. The CLS2 lure ws not included in this experiment ecuse of indequte vilility of lures. A trp without lure (lnk trp) served s control. In ech lock, six trps with the different commercil pheromones nd lnk trp were plced on the floor rndomly, on line, 1.5 m from ech other nd 2 m wy from the wlls. Four locks were deployed in the min re of the sement nd the fifth lock ws the smller inner room locted within the sement. Lortory-rered L. serricorne were relesed in the sites to estlish popultions for the trpping experiments. Detils of the protocol used for rering L. serricorne used in this nd susequent experiments re provided in previous report (Mhroof nd Phillips 27). To estlish L. serricorne popultion for trpping, we plced two jrs contining different life stges of L. serricorne plced in two rndomly selected loctions, hlfwy etween the trps nd 2 m wy from the wlls. Ech jr contined pproximtely 2 dult L. serricorne. Insect jrs were replced with fresh colony twice during the experiment, fter 3 nd 6 weeks from the eginning of the experiment. The trps were inspected weekly nd the numer of L. serricorne cught ws recorded. The trps were monitored for 9 weeks during which time trps with pheromone lures were rotted weekly to different positions, nd trps were replced every 3 weeks. Two HOBO Ò dt-logging units (Onset Computer Corportion, Bourne, MA) were used to mesure environmentl conditions t SPREC nd ASB. One light ul (75 W; high-pressure sodium lmp) ws kept illuminted throughout the dy nd night in the multipurpose room t SPREC in order to comply with the OSU sfety requirements. The sement of ASB ws kept totlly drk except when the trps J. Appl. Entomol. 132 (28) 161 167 ª 28 The Authors Journl compiltion ª 28 Blckwell Verlg, Berlin 163
Evlution of ttrctnts for stored-product Anoiide R. M. Mhroof nd T. W. Phillips were checked nd serviced weekly. The men temperture, reltive humidity (RH), nd light intensity (LI) t SPREC verged from two HOBOÒ units for 9 weeks were 3..1 C, 52.3.5%, 5.4 2. lux (n = 761) respectively. Men temperture nd RH t ASB during the study period were 26..1 C nd 56.6.1% respectively. Responses of L. serricorne to pheromone nd host-plnt voltiles Experiments were lso conducted t SPREC from Jnury to Mrch 26 to investigte responses of L. serricorne to StorgrdÒ II sticky trps ited with pheromone lures lone, pheromone plus host-plnt voltiles or host-plnt voltiles lone. The plnt mterils used in the study ws dried red chilli (C. frutescens, Dicotyledon: Solncee; Bolner s Fiest Product Inc., Sn Antonio, TX). There were six experimentl tretments: the first consisted of trps ited with 1 mg of synthetic pheromone lures lone. The lures used in the study were fricted in our lortory. Briefly, synthetic serricornin (nonn-3-one, 4-6- dimethyl-7-hydroxy; 95% pure; Bedoukin Reserch, Inc., Dnury, CT) ws diluted in doule-distilled hexne (Regent grde ACS; Phrmco, Brookfield, CT) to finl concentrtion of.2 mg/ll, of which 5 ll of the pheromone solution ws impregnted into ruer sept (No. 11.5; Fisher, Pittsurgh, PA). The ruer sept were first clened y soking in hexne for 12 h, then in dichloromethne for 12 h, nd then drying in fume hood for 24 h. The second tretment consisted of trps ited with 5 g of ground dried red chilli pcked in plstic meshed pouch (5 cm long, 3.7 cm wide nd 3. mm mesh). Chilli peppers were pulverized with mechnicl grinder, nd sifted using n 85-lm meshed sieve (U.S. No. 2; Seeduro Equipment Co., Chicgo, IL). In third tretment, we pplied 5 ml of filtered chilli extrcts, t concentrtion of 1 g equivlent per ml of hexne, to no. 3 cotton dentl roll (3.75 cm long nd 1.25 cm wide) (Tidi Brnd, Neenh, WI). The impregnted dentl wicks were plced in the middle floor of the sticky trps. The chilli extrcts used in the study were extrcted with doule-distilled n-hexne s descried y Mhroof nd Phillips (27). The fourth nd fifth tretments consisted of trps ited with 5 g of chilli powder plus 1 mg of synthetic serricornin nd trps ited with 5 ml of chilli extrct plus 1 mg serricornin respectively. Blnk trps nd trps ited with wicks loded with 5 ml of n-hexne served s controls. Trps were rrnged in rndomized complete lock design with four locks, ech consisting of five 164 tretments nd two controls rrnged rndomly. Trps were suspended verticlly on string, 1.7 m off the floor nd t distnce of 2. m from the side wlls, with out 1.5 m etween trps. Lortoryrered L. serricorne were relesed every 3 weeks, s descried ove. Trps were checked every week t which time the numer of insects cught per trp ws recorded, nd ech trp rotted clockwise. All lures nd trps were replced every 3 weeks. In the sttisticl nlysis we included only 6 weeks of trp ctch dt. Dt from week 5, 7 nd 9 were not included in the nlyses ecuse trp ctches were nil during these 3 weeks. Men temperture nd RH during the study period were 27.3..4 C (n = 75) nd 5.6.3% (n = 75) respectively. Men LI ws 5.3 1.8 lux (n = 761). Sttisticl nlyses Experiments were conducted s rndomized complete lock design, with lure types serving s tretments nd replictions serving s locks. Dt from ech lock within two trpping weeks for S. pniceum nd within trpping week for L. serricorne were pooled prior to nlyses. Time (weekly or iweekly recordings) ws considered s rndom effect nd tretments s fixed effects. Prior to dt nlysis, count dt were trnsformed to log 1 (X + 1) scle to stisfy the ssumption of normlity nd homogeneity of vrince (Zr 1984). Dt were sujected to nov y using the mixed procedure of ss (SAS Institute 23). lsmens were used for seprting tretment men vlues t =.5 level. Actul men vlues nd SEMs re presented in the text nd figures. Results Trpping S. pniceum using different commercilly fricted stegoinone Significnt differences (F = 5.49; d.f. = 4, 14; P =.72) were oserved etween men numer of S. pniceum ctches in trps comprising vrious tretments in the retil pet stores. With the exception of CSP2, the numer of S. pniceum cptured in trps ited with ny of the three types of CSP1 lures ws similr to oservtions in united trps (fig. 1). Trpping L. serricorne using different commercilly fricted serricornin Significnt differences (F = 4.6; d.f. = 3, 31; P =.89) were oserved etween numer of J. Appl. Entomol. 132 (28) 161 167ª 28 The Authors Journl compiltion ª 28 Blckwell Verlg, Berlin
R. M. Mhroof nd T. W. Phillips Evlution of ttrctnts for stored-product Anoiide Men (+ SE) numer of S. pniceum 18 16 14 12 1 8 6 4 2 CSP1 type 1 CSP1 type 2 Fig. 1 Men numer (SE) of dult S. pniceum cptured in retil pet supply stores in trps ited with different commercil frictions of stegoinone. Men vlues were sed on n = 4. Men numers with different letters re significntly different (P.5, LS mens). CSP1 type 3 CSP2 Blnk Men (+ SE) numer of L. serricorne 35 3 25 2 15 1 5 CLS1 CLS2 Fig. 2 Men numer (SE) of dult L. serricorne cptured t the Stored Products Reserch nd Eduction Center t Oklhom Stte University in trps ited with different commercil frictions of serricornin. Men vlues were sed on n = 9. Men numers with different letters re significntly different (P.5, LS mens). CLS3 Blnk L. serricorne cptured in the pheromone trps compred with the lnk trps t SPREC. Pheromoneited trps cptured, on verge, eight times more L. serricorne thn the lnk trps (fig. 2). However, the numer of eetles cptured in trps ited with CLS1, CLS2 or CLS3 ws not significntly different (P >.5) from ech other. Similr to oservtions t SPREC, the numer of L. serricorne cptured t ASB were significntly (F = 8.4; d.f. = 6, 55; P =.1) higher in pheromone-ited trps thn in lnk trps. The results lso showed tht trps ited with CLS5 type1 nd CLS type 3 cptured the highest nd lowest numers of L. serricorne, respectively; nd trp ctches etween these two lure types were significntly different from ech other (fig. 3). Responses of L. serricorne to pheromone nd host-plnt voltiles The trpping experiment conducted t SPREC comining pheromones with host odours showed significnt differences (F = 4.21; d.f. = 6, 3; P =.34) mong tretments (fig. 4). Trps ited with serricornin plus red chilli voltiles cptured out three to four times more eetles thn trps ited with pheromone or chilli voltiles lone. An verge of one to two eetles per week were cptured in trps ited with pheromone, chilli powder or chilli extrct lone. Sttisticl nlysis confirmed tht the numer of L. serricorne cptured ws not significntly different (P >.5) mong the three tretments. Men (+ SE) numer of L. serricorne 18 16 14 12 1 8 6 4 2 Discussion CLS1 CLS3 CLS4 Fig. 3 Men numer (SE) of dult L. serricorne cptured t the Animl Science Building t Oklhom Stte University in trps ited with different commercil frictions of serricornin. Men vlues were sed on n = 9. Men numers with different letters re significntly different (P.5, LS mens). We noted significnt vrition in the efficcy of S. pniceum lures tht re commercilly ville or products tht re in the process of development. CSP2 is reltively etter lure to trp S. pniceum ecuse of the reltively high numer of dults cught in trps with CSP2. If the pheromone lures re ttrctive, then lures drw insects from distnce, nd insects will e trpped rther rpidly in high numers. It is noteworthy to stte tht only CLS5 type 1 CLS5 type 2 CLS5 type 3 c Blnk J. Appl. Entomol. 132 (28) 161 167 ª 28 The Authors Journl compiltion ª 28 Blckwell Verlg, Berlin 165
Evlution of ttrctnts for stored-product Anoiide R. M. Mhroof nd T. W. Phillips Men (+ SE) numer of L. serricorne 5 4 3 2 1 c Blnk c n-hexne c Pheromone Fig. 4 Men numer (SE) of dult L. serricorne cptured t the Stored Products Reserch nd Eduction Center t Oklhom Stte University in trps ited with serricornin lone, Cpsicum products lone, or in comintions. Men vlues were sed on n = 6. Men numers with different letters re significntly different (P.5, LS mens). very few rnds of commercilly fricted stegoinone re currently ville in the mrket for monitoring nd s result monitoring nd trpping of S. pniceum re reltively limited when compred with L. serricorne monitoring. Our studies showed tht commercil pheromone lures from different mnufcturers, or lures mde in our lortory using the technicl grde serricornin, re effective in monitoring popultions of L. serricorne. Synthetic serricornin used y different mnufcturers seems eqully ttrctive for given type of trp design. Vritions oserved in response of L. serricorne to sex pheromone lures etween SPREC nd ASB my e due to vritions in the trp designs, eetle popultions relesed, or environmentl conditions experienced etween the two loctions. Movement nd flight of insects were influenced y the indoor mient temperture, reltive humidity conditions nd light vilility (Hgstrum et l. 1996; Tremterr nd Scirrett 24), nd therefore vrition in mient conditions t SPREC nd ASB my hve ffected the trp ctches etween the two experiments. It is known fct tht the efficcy of pheromone lures my e enhnced y comintion with host odours (Chmers 199; Lndolt nd Phillips 1997). 166 c Chili powder c Chili extrct Pheromone+chili powder Pheromone+chili extrct Results from the experiments in which we ited trps with synthetic serricornin lone, pheromone plus voltiles from red chilli, or chilli voltiles lone showed tht trps ited with pheromone plus chilli voltiles were significntly more ttrctive to L. serricorne thn trps ited with synthetic pheromone or chilli voltiles lone. To our knowledge this is the first pulished report indicting synergistic effect of synthetic pheromone nd plnt voltiles. These findings could help complement the current mngement progrmme for L. serricorne y exploiting the synergistic effects of comining synthetic pheromones with plnt-derived voltiles to improve mss trpping of the pest or to improve the sensitivity of the trp. Greter trpping sensitivity might e more of n dvntge when chnces of ctching insects re low. The use of n ttrctnt in trps most often improves trp performnce y incresing the chnce of detecting low-density insect infesttions, e.g. in loclities where the pest hs not yet estlished itself nd the popultion is still low or where the insect ctivity is suspected ut not yet detected. In this sitution n improved pheromone lure y comining host plnt-derived voltile my sustntilly increse the chnces of detecting L. serricorne. Our results showed further tht, specificlly the chilli extrct when comined with the pheromone, ws more ttrctive thn the chilli powder lone or chilli extrct lone, nd suggest tht trps with host mterils extrcted with suitle solvent to concentrte voltile compounds, nd then comined with the pheromone my e lterntive thn the fresh host mterils or their extrcts lone. In summry, our studies clerly showed tht vrious types of lures loded with serricornin re ttrctive to L. serricorne, ut not ll lures loded with stegoinone re ttrctive to S. pniceum. There is potentil for improving lures for S. pniceum, nd for gining improved trp sensitivity for L. serricorne y comining host voltiles like those from Cpsicum spp. Acknowledgements We cknowledge severl commercil vendors for providing Anoiide sex pheromone lures nd trps, nd pprecite Mrgie D. Brton for her technicl ssistnce during the S. pniceum trils. We thnk Drs Xingwei Hou nd Crmen Greenwood for their comments on the erlier drft of this mnuscript. We specilly cknowledge two nonymous reviewers for their constructive comments to improve this mnuscript. J. Appl. Entomol. 132 (28) 161 167ª 28 The Authors Journl compiltion ª 28 Blckwell Verlg, Berlin
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