is )-2,13- )-2,13- Z Z , , E Z O. sacchari O. sacchari (Hollingsworth than ( gluresof( )-2,13-octadecadienal µ Z a , )13-octadecadienol, was can be a direct pest and an Z Z is federally regulated as a 5 O. sacchari 2010 Society of Chemical Industry malesandcanbeusedasa rambutan, mangoes and sweet c 7 O. sacchari O. sacchari O. sacchari include palms and Therefore, 6 O. sacchari Correspondence to: Matthew S Siderhurst, EasternPark Mennonite Rd, University, Harrisonburg, 1200 VA 22802, USA. E-mail: [email protected] US Pacific Basin Agricultural Research Center, USDA-ARS, Hilo, HI, USA DepartmentofChemistry,EasternMennoniteUniversity,Harrisonburg,VA,USA In the United States, ∗ a b RG, unpublished data), banana, indirect pest on horticultural commodities exportedbecause from of Hawaii, the riskby of quarantine commodity inspectors. rejectionto Commodities if the larvae exported US are fromwith mainland found Hawaii that are sometimes found to be infested would provide a non-destructive population-monitoring tool. This attractant could be used to studyof the seasonality banana and distribution in various host crops, and to facilitate timing potatoes (Folletttechniques P, for private monitoringsampling for banana communication, larvae through 2009). destructiveIdentification populations assessment of of Current require the a crop. sex pheromone attractant for quarantine pest. responsible for serious damagegirdling to following pruning coffee activities. trees, caused by stem otherwise desirable horticultural traits. In addition, aldehyde) but did not elicit responses from antennae of male y-mass spectrometry. An analog, 2/3,( se compounds when the corresponding stereoisomeric alcohol b Robert G Hollingsworth, could offer new methods for detection, study and control. ∗ Af- Cy- b elicited responses from antennae of male moths. This compound was In the 1,2 3 Bojer, is a polyphagous agricultural pest in many tropical areas of the and )-2,13-octadecadienal attracted more male Z O. sacchari , E O. sacchari 3 Bojer (: ), palms, and Elisa J Troyer b ; banana moth; attractant; octadecadienal is expanding its host range in Hawaii. is recorded only from Florida and Hawaii, Bojer (Lepidoptera: Tineidae): Matthew S Siderhurst, )13-octadecadienal by gas chromatograph : 454–460 www.soci.org )-2,13-octadecadienal has been identified as an attractant for Z isageneralistfeeder,andinfestationstypically a Z 66 Opogonasacchari , E O. sacchari O. sacchari 2010; Larvae usually feed internally, burrowing within the 2 Opogona sacchari . with such severe damage to some varieties that they sp. (Hollingsworth RG, personal observation). There is )-2,13-octadecadienal. In field trials, (

4 Z , E

2010 Society of Chemical Industry Opogonasacchari c octadecadienal caught as many males as did traps holding virgin females. CONCLUSION: ( monitoring lure of populations of this moth. Keywords: United States, and has been in Hawaii since 1990 or before. stem or roots of host plants.from This the Mascarene Islands was in originally the described Indianduring Ocean the in last 1856. 30 However, years, itand has subtropical been areas reported in from Africa, humid Europe and tropical South America. begin on diseased, damaged ormaterial dead associated tissue, with or the on croplarvae other or organic can commodity. However, burrow older intoDracaena healthy plant tissues, such as stems of evidence that During the past 8 years,on it orchids has been and noted coffeehigh as seedlings in a organic being root/stem matter grown feeder (HollingsworthExtensive in RG, damage potting personal to observation). pineapple media plantings hasrecently, also been recorded Pest Manag Sci is an agricultural pest in many tropical areas of the world. 1INTRODUCTION The banana moth, BACKGROUND: The banana moth, Abstract sacchari Eric B Jang, David N Showalter world. The identification of an attractant for male Sex attractant for the banana moth, provisional identification and field evaluation (www.interscience.wiley.com) DOI 10.1002/ps.1922 Research Article Received: 2 July 2009 Revised: 27 November 2009 Accepted: 21 December 2009 Published online in Wiley Interscience: 9 February 2010 RESULTS: A compound extracted from female also detected in some extractsmoths. at Electroantennograms roughly a of 1 synthetic :and 20 candidate ratio ( (alcohol dienals : foundoctadecadienal. the Attraction strongest was not responses improvedwas from for added ( either at of ratios the of 1 : 1, 1 : 10 or 1 : 100 (alcohol : aldehyde). Jackson sticky traps containing 250 fected crops include banana, coffee, pineapple,maize, papaya, sugarcane, bamboo, stored tubers andDracaena, ornamental crops, , including cas revolute are now avoided by the pineapple industry in spite of their identified as a 2/3,(

454 455 ). × m 1 µ − )and 25 1 . − , positive × 0.25 mm ID, 0 C respectively. The oven × ◦ C). The injector, in splitless was used to test the system. C for 1 min, and was then C, using helium as a carrier ◦ ◦ ,witha1minstartdelaywith 8 ◦ , 1 − C and was held for 13 min. Whole ). The order of the aldehydes was ◦ × Cmin ◦ ), which were combined with a Y-type 1 O. sacchari − www.interscience.wiley.com/journal/ps to 240 1 − Cat10 m film thickness). The temperature program ◦ ). µ C, using helium as a carrier gas (1.1 mL min 1 ◦ − 25 Cmin , with a 1 min start delay with the injector temper- . ◦ 1 , negative control), 1-hexanol (1–6:OH, 3 − )and1–6:OH(3 × × Cmin ¨ ulheim an der Ruhr, Germany) was attached to the base of ◦ Electroantennograms of single synthetic aldehyde compounds www.soci.org H NMR spectra were obtained with a Bruker DRX-400 FT- film thickness) with helium as carrier gas (2.3 mL min the connector, and the effluentionization was detector split 1 (FID) : 1 and(EAD) between via the the a flame electroantennogram heated detector transfer line (250 used was 60 to 250 temperature program began at 80 gas (1.0 mL min the injector temperature set at 250 make-up gas (10 mL min control), three sequentialair puffs only of (3 each of the test aldehydes, were recorded usingHowever, these the recordings were recording takenintroduced into system with the compounds humidified described directly airstreampreparation. leading above. This to was the accomplished antennal by puffingPasteur air pipet through containing a the glass compound of interestAll on replicates filter paper. consisted of sequentialair puffs in only the (3 following order: moth heads or excised antennae were(Spectra secured 360; with Parker electrode gel Laboratories, Inc.,parallel Fairfield, metal paddle NJ) electrodes between of the aholder Syntech (5 EAG mm probe antenna separationpassed between over paddles). the Humidified antennal aireffluent preparation from was the and EAD acted transfer line. as Theby antennal a signal the carrier generated EAD for high-impedance was amplifier and amplified analyzed with and theSyntech FID filtered GC-EAD2000 signal software. by using Z11-16:Ald, aelicit previously EAG Syntech responses shown from to NL 1200 2.4 Electroantennography Electroantennographic responses weregas recorded chromatograph–electroantennogram either coupled withEAD) system a or (GC- withelectroantennographic an responses electroantennogram to (EAG)extracts female set-up were pheromone only. gland recordedgas Male chromatograph using coupled an todetector a Agilent system Syntech Technologiesequipped electroantennogram (Hilversum, with 6890 The an HP-5 Netherlands). column The (30 m GC was ature set at 250 1 at 10 connector. A Graphpack-3D/2 flowKG, splitter M (Gerstel GmbH & Co. The second instrument, located in Virginia,G1800A was a GCD Hewlett system Packard equipped with0.25 an mm HP-5 MS ID, column 0 (30 m ramped at 10 mode, and FID were held at 250 and 275 NMR spectrometer equipped withAll a broadband spectra gradient were probe. tetramethylsilane (TMS) recorded as an in internal standard. deuterated chloroform with2.6 1% Chemicals All solvents and reagent compounds were purchasedAldrich, from Sigma- St Louis, Missouri or Acros Organics, Geel, Belgium. All randomized in each replicate.the Aldehyde filter paper (100 and ng) allowed was 30to s evaporate dosed for before on the puff dichloromethane dispensing. solvent Onlyresponses to replicates 1–6:OH that showed were included in the analysis. 2.5 Nuclear magnetic resonance C )- ◦ Z 9,10 . that used 8 et al C while ◦ O. sacchari 2010 Society of Chemical Industry 70 m film thick- µ c − O. sacchari ) contained the 25 . 20 . and Ioneda The majority of the 8 in greenhouses over a Cryptophlebia batrachopa 12–19 spp.toa99 : 1mixtureof( . O. sacchari 0.25 mm ID, 0 )13-octadecadienyl compounds, Z × O. sacchari Opogona O. sacchari sp. sex pheromones have not been : 454–460 66 These results clearly indicate the existence of a )-8-dodecenylacetateswas,however,notedduring pheromone is similar in structure. The objective of Opogona sacchari 2 E 2010; Opogona In Florida, traps baited with virgin females were used 11 )-11-hexadecenal (Z11-16:Ald) elicited responses from moth Z O. sacchari Although Early work to investigate a possible pheromone for Attractant for 1 year period. identified pheromones are 2/3,( predominantly alcohols, aldehydesmoths and acetates. attracted Althoughsubfamilies the to within these Tineidae,the it pheromones is belong reasonablethe present to study to was to postulate characterize, synthesize different pheromone and that attractants field for test sex 2.3 Gas chromatography–massExtracts spectrometry from calling femalematography–mass moths spectrometry were (GC-MS) using analyzed twoThe by instruments. first, gas located chro- in Hawaii,6890N consisted gas of chromatograph an interfaced Agilent tomass Technologies a Hewlett-Packard selective 5973 detector equippedDB-255 MS with column (both either 30 m an HP-5 MS or a exhibiting calling behavior 2–4 h prior toAbdominal the tips onset were of subsequently photophase. excised and extracted for 5in hexane. min Hexane extracts were then transferred intovials conical glass and evaporatively concentratedstream. under a purified nitrogen ness). The standard temperature program used was 80 to 240 identified,pheromonesforothertineidmothshavebeenidentified and utilized for purposes of control. female-produced sex pheromone in Pest Manag Sci 22.1 MATERIALS AND METHODS Insects used in tests were taken from a colony of was undertaken by Rotundo and Tremblay had been in continuous laboratorycollected from culture infested since rambutan 1999, bark originally incolony Kurtistown, is Hawaii. maintained The within ventilatedage plastic colony) containers provisioned (mixed- with artificial(continuous diet on production an as-needed cycle). basis based The diet artificial (modified bean- from and Follett wheat- and Lower Both groups found that male mothsto were extracts behaviorally from responsive calling females. However, neither groupthe identified active pheromone components. Rotundo and Tremblay of pesticide applications with peak pestpheromone-based abundance. tools Additionally, such as pheromoneused disruption as could part of be a control program for this pest. 2.2 Gland extractions Female moths, 2–3 days old, were frozen at following proportions of blended ingredients: 468 gnorthern beans, cooked 120 great g wheat meal, 70acid, g yeast flakes, 4.4 7.0 g g ascorbic sodium1400 mL hot benzoate, water. 2.2 g sorbic acid, 40 g agar and electroantennography (EAG) to assay severalto compounds be known pheromone componentsthat of ( other Lepidoptera, andantennae, found but fieldpublished.Fieldattractionof work described in their paper was never to monitor populations of a study in Malawi with the tortricid 8-dodecenyl/( (Meyrick). . 13-, 1m gof gof et al Z µ µ ∼ 12 cm 2, E × : 454–460 13-18:Ald. Z 66 , 13-18:OH and 13-18:Ald and E2 Z Z , 2010; 3, 6 months prior to E E2 < 4 grid of traps (6.3 m × to red rubber septa (5 mm; 2 Pest Manag Sci Cl 2 13-18:Ald. Thus each trap holding a Z , 13-octadecadienyl aldehydes, acetates Z E2 13-18:Ald plus an additional amount (250, 3, Z Z 2, 40 m distant from the closest traps comprising Z ∼ 13 and 5 m away and baited with the artificial attractant. While Z were counted every 3–4 days. Sticky inserts and virgin . g) of its corresponding alcohol. 5 grid of traps (also spaced 6.3 m apart) baited with lures 4 2, µ Z × ∼ 5 . 4 traps holding virgin females was set up in the other half of 13-18:Ald) when these were used singly or blended with 13-18:OH were tested in only one of the four experimental 13, 13-18:Ald or × 5 m above the ground in a double-row pattern, with each row Z Z Z Z . In a second papaya field under commercial production located , 1 2, 3, 3, tests, and, because of the rocky soil, some treesproviding fell breeding over material and for rotted, the banana moths collectedtraps. in the Efficacy of massin Jackson trapping traps was holding assessed virginpreviously females, described, by except set all moth up Jackson traps and capture were attached monitored as Hawaii (Hawaii Island), in 2007were and sparse, 2008. Surviving and papayaweeds. trees Banana the moths in ground this fielddying was were papaya trunks. reproducing covered Jackson on sticky dead cardboard with and trapsManufacturing, Fresno, (Better CA) low-growing World were used to trap male moths.compounds Test lure were applied in CH Papaya trees in this field3.15 were m about apart. 2–3 Trees m had in begun height, bearing with fruit rows above the ground on bamboo stakes.spacing) A holding 4 virgin females waslarger positioned 5 at the center ofholding a 50 mg each of virgin female was surrounded onlocated four sides by artificial lurevirgin traps females were replaced every 3–4the days, beginning septa of were the dosed at trialcollection and were period. not As replaced an overof experimental the 4 2 control, week an additionalthe grid papaya field the first grid. 2.9 Analysis EAG results were analyzed using ANOVAof followed means by comparison using the Student–Newman–Keuls test. All analyses less than 1 km away, aout mass trapping in trial (field 2009 test 3) using was red carried rubber septa baited with E Z their corresponding alcohol at an1 : aldehyde/alcohol 10 ratio or of 1 : 1the 100. : specified 1, All chemical. Septa septa holding using blendsE2 contained single 250 attractants held 250 25 or 2 and alcohols. Because of technicalZ difficulties, replications. Therefore, theseanalyses, compounds but results were are reported. excluded Fieldof test from 2 males measured to response the two best-performing lures ( Kimble/Kontes, Vineland, NJ), andsticky the inserts septa of were Jackson traps. placed Invirgin on traps female used the (2–3 as days positive old) controls,aluminum was a confined within window-screen a custom-made, enclosurelong) (3 attached cm to the diameter was ceiling supplied of to the females trapglued by along to wetting its the dental inside length. wicks of Water ∼ the that enclosure. had Traps been were hungholding a from complete trees set of randomized treatments.rows Spacing was within 15–20 m,O. and sacchari rows were separatedfemales by were replaced 30 at m. that time. Males Forfour each of trapping experiment, replications wereranging carried from 7 to out 18 days over inNew duration time, (2–5 traps, trapping with periods new each). replications septaused and in a each newof replication. males randomization Field to scheme test chemical were Twelve different lures 1 compounds were when tested, measured comprising compounds the the were attraction used singly. 2 2 in E Z 13- 13- 13- 13- )to 98% E Z Z Z 2 > www.soci.org EB Jang (1 mL) )-3,13- )-2,13- 2, 2, 3, 2, Cl Z Z 2 E E Z E 2 , , ) to yield E E 2 13-18:Ald, 13-18:OH) Cl (2 mL) was 2 Z Z Cl 2010 Society of Chemical Industry HNMRalde- 2 2 1 2, 2, c E Cl Z GC-FIDanalysis 13-18:OAc) and 2 H NMR analyses Z solution (15 mL). 22 1 3, 3 and concentrated Z 13-18:OH) were gifts 4 )-2,13-Octadecadienal Z )-2,13-Octadecadienol Z Z All acetates were , 3, , 13-18:Ald) and ( 13-18:OH by Dess–Martin )-2,13-Octadecadienyl ac- E Z Z 13-18:OAc) were prepared 23 Z Z Z Z L, 2.1 mmol) and pyridine , µ 3, 2, . The resulting oil was puri- Z H NMR analysis. 3, 1 Z Z E )-2,13-Octadecadienal ( )-3,13-Octadecadienol ( 13-18:Ald isomerizations to Z 90% pure by GC-FID analysis. Z , Z , > E E 3, Z in vacuo )-2,13-octadecadienyl acetate ( )-2,13-octadecadienol ( hydrochloric acid (5 mL) and extracted Z 98% pure by GC-FID analysis. ( Z , , M E > Z (4 mL) and stirred at room temperature 13-18:Ald by 13-18:Ald) were prepared from their corre- Z (100% hexanes to 1 : 1 to 100% CH 2 Z (5 mL), dried over MgSO 5 mL). The organic extract was washed with 3, 2 Cl 3, 3 2 Z (100% hexanes to 1 : 1 to 100% CH E Cl )-3,13-octadecadienol ( × 13-18:OH (166 mg, 0.62 mmol) in CH 13-dienyl compounds known to have pheromonal 2 2 Z Z Z , Cl )-3,13-octadecadienyl acetate ( Z 13-18:Ald and 2 2, )-3,13-octadecadienal ( Z 2 aldehyde, indicating a slight isomerization of the Z The mixture was then diluted with diethyl ether , 13-18:OAc), ( Z Z E , Z Z 3, Z and concentrated E 21 95% pure by GC-FID analysis. ( 2, 13-18:Ald showed trace contamination (4%) of the corre- 4 E Z . The resulting oil was purified by column chromatography > 13-18:Ald) was prepared from 13-18:OH) and ( 13-18:Ald (118 mg, 72%). ( 13-18:Ald or Z )-3,13-octadecadienyl acetate ( 2, Z SO Z Z Z 2 Z 2, 2, , 2, 3, E E Z 18:OH) and ( saturated NaHCO hydespectramatchedthosepreviouslyreported. provide the desired aldehyde (71–79% yield). 18:Ald, showed less than 5% contaminants for each aldehyde ( E double bond. Isomerization was not detected with 18:Ald), ( of sponding Z 2.8 Field trials Single candidate attractantsfield and tested several binary in mixtures an were abandoned papaya plantation near Keaau, pure by GC-FID analysis. was added to a1.0 mmol) solution in of Dess–Martin CH periodinane (419 mg, vacuo to yield the desiredwere acetate confirmed by (67–73% GC-MSously yield). analysis reported Acetate fragmentation and patterns. structures comparison with previ- www.interscience.wiley.com/journal/ps solvents were HPLC grade. ( from Scentry Biologicals, Inc., Billings, Montana.were Both compounds stirred at room temperature forpurified 4 by h. column The chromatography crude on mixtureof silica was gel hexanes then : with CH a gradient (20 mL) and treated with saturated NaHCO with hexane (3 The organic layer wasNa removed, washed with water, dried over for 3 h. fied by column chromatographyhexanes on : CH silica gel with a gradient of were 88, 16 and 19% respectively). ( etate ( sponding alcohols by oxidation with(PCC). pyridinium A chlorochromate solution of0.38 mmol) one and of PCC the (200 three mg, octadecadienols 0.93 mmol) (100 mg, in CH octadecadienal ( ( were purchased from Pherobank, Wageningen,Both the compounds were Netherlands. 18:OAc), ( activity in Tineidaetrophysiological were synthesized and to facilitate field further studies. elec- ( 2.7 Synthesis Candidate 2/3, Octadecadienal used in field test 3Wageningen, was the obtained from Netherlands, Pherobank, ( from their corresponding alcohols by esterification withhydride. acetic A an- solution of one0.38 of mmol), the three acetic octadecadienols (100 anhydride(1 mg, mL) (200 was stirredwas at poured room into temperature cold 1 overnight. The mixture ( oxidation.

456 457 13-18:OH Z 13-18:Ald or 3, Z Range over Z 2, trapping periods Z Opogona sacchari 13-18:Ald (0.70 MTD), the Tukey Z grouping males caught in Jackson sticky 2, 13-18:OH and 2)2) A A 6)3) B B 04)4) A B 04)4) A B Z ...... Z plicates consisted of sequential pairs of traps monitored every 0 0 0 0 0 0 0 0 3, ± ± ± ± ± ± ± ± E SE) (mV) SNK grouping 01)01)01)01) B00) B B B 0.0–0.5 B 0.0–0.3 0.0–0.3 0.0–0.3 0.0–0.0 05) B 0.0–1.7 21)20) A A 0.0–9.0 0.0–4.5 49)18)24)06) A B B B 0.0–5.0 0.0–3.3 0.0–1.8 0.0–0.8 ...... ± SEM) 0 0 0 0 0 0 0 0 0 0 0 0 ( ± ± ± ± ± ± ± ± ± ± ± ± ± 0.00.0 – – – – www.interscience.wiley.com/journal/ps day ( Opogona sacchari Electroantennogram response a Males trapped per 80) (Table 2). Other single-component chemical 13-18:Ald (1.69 MTD). E2,Z13-18:Ald caught more Z b b = 2, 05) Electroantennogram responses of male Number of E . 0 < 05, df gofchemical) . 13-18:OH13-18:OH 0.01 ( 13-18:OAc 0.01 ( 13-18:Ald 1.2 ( 13-18:Ald 3.1 ( 13-18:Ald13-18:Ald 0.70 ( 13-18:OAc 0.36 ( 0.08 ( P 13-18:OAc13-18:OAc 0.01 ( 13-18:OH 0.01 ( 0.00 ( 13-18:Ald 0.8 ( 13-18:Ald 2.5 ( 13-18:OH 13-18:Ald 1.69 ( 13-18:Ald 0.16 ( 2 week exposure period, no male moths were collected in µ 0 Z Z Z Z Z Z Z Z Z Z Z Z Z Z Z Z ∼ Results shown for comparison purposes only. These chemicals were Attractants were assayed using 2, 3, 3, 3, 2, 2, 3, 2, 2, 3, 2, 3, 2, 3, 2, 3, = E E Z E Z E Air only1-6:OH 0.34 ( 2.6 ( Z E Z E tested in only one replication. moths to candidatepuffs in attractants. the Re order shown,randomized except in each that replicate. the Air order was1-hexanol used of was at the used a as aldehydes negative a control, was while positivethat showed control. responses Results to from 1-hexanol are ten shown. replications same Means followed by letter the aretest; not significantly different (Student–Newman–Keuls Treatment Treatment Virgin female (A)Virgin female (B)E 1.82 ( 1.75 ( a b Table 1. Air only1-6:OHZ 0.33 ( 2.7 ( Table 2. traps baited with single-component chemical lures250 (septa loaded with Z Z E Z 3–4 days over 2–5 trappingwere periods carried (one replication). out Four over replications time. www.soci.org lures tested caught about half as manyless. moths As as mentioned previously, were tested in only one of the fourthe experimental replications. Over Best catch was obtained in the two experimental controls (identical treatments) using virgin femalescontaining (1.75–1.82 MTD)than and twice as many in moths as did traps the next best attractant. This differenceP was significant (Tukey’s HSD, any of the four trapsFor all associated attractants, with temporal these variation in treatmentshigh. trap (Table catch For 2). was example, extremely in the a maximum number trap of during moths any captured of the 3–4 day trapping periods was 13 21, 03, . Z )13- 23,25 0 Z = 3, 1, 80). / F < 2 = 22 P 13-18:Ald Z Using the Z 2010 Society of Chemical Industry 24 0001, . c 13-18:OH) 08, df 0 . Z 0 < = P 13-18:Ald, electroan- F Z 05 level. Field data on abdominal tip extracts 8, . . 0 0 The average catch of males < = An EAG inactive compound P P and a fragmentation pattern 13-18:Ald. 25 Z z / 2, O. sacchari E m )13-octadecadienol (2/3, Z 13-18:Ald). : 454–460 13-18:Ald on both a non-polar HP-5 MS and Z B’ in trap catch was then computed for all Z 66 − 2, 05) was used to compare means. . E 0 Opogona sacchari 13-18:Ald and 2010; = 3,80)andtreatments(luretypes)( Z 2, = α Z and a fragmentation pattern consistent with a 2/3,( 13 geometric isomer or those containing the z 1,df / Z . 11, 80) but not trapping row ( 3 m 3, = Given the isomerization problems of 2/3, Field test 1 (compounds tested singly). / = 2 Attractant for df octadecadienal (2/3, F Pest Manag Sci revealed aantennae single of compoundshowed male that that moths. the elicited264 GC-MS EAG analysis responses active of from compound these had extracts a mass ion of matched that of a polar DB-255 MS column, precisebecause identification became many difficult ofduring these GC analysis aldehydes (even with are cool on-column unstable injection). and isomerize was also intermittently detectedlimits in of a range theThe near instrument two the detection compounds and(alcohol : aldehyde). were was While the retention detected time not of the roughly 2/3, definitively in identified. a 1 : 20 ratio Coupled gas(GC-EAD) chromatography–electroantennogram analysis of detection female 3RESULTS of significance were made at the male trap capturesby were the normalized number by ofmales dividing days caught trap in per captures the dayperiods trapping comprising in each period. each replication, The was trap,variable used number averaged in as of over the ANOVA dependent all models,treatment (lure trapping while type) were replication, used trapping as independentHSD row variables. Tukey’s test and ( with a mass ion of 266 same ANOVA model, fieldlure types test had 2 been recategorized dataE as were either those also containing analyzed the after isomer (with experimentalCasual controls observations deleted suggested from thathigh a number the of particular moths dataset). trap over one catchinghighnumbersofmothsoversubsequentsamplingperiods,relative sampling period a tended to catch to moth capture in nearbyTo traps test baited with this the hypothesis, sametrap the attractant. catch between authors pairs calculated of traps thefor each baited difference trapping with period in the (3–4 same attractant daystest duration) 1 using data associated from with field themost three moths. artificial attractants Within catching eachtrapping the periods), trapping the trap associated replication with the (comprising highestwas moth 2–5 designated catch as ‘A’,the and the difference other ‘A traptrapping was periods, designated except as for ‘B’; thethe trapping highest period trap catch. associated The with 95%difference was confidence used interval as for the the significance mean include test: if zero, the this interval did waslocation not of considered an as individual trap evidencetrap was catch. that an important the variable specific affecting tennograms were recorded with syntheticseparation. aldehydes without The GC resultsWhen of grouped, this the 2,13-dienals testing elicitedthe are stronger 3,13-dienals. responses There displayed was than not in a significant differencebetween Table in 1. response per trap per dayfrom (MTD) 1.82 to in 0.008 glue (Tabledifferences traps in 2). male ANOVA capture across associated results treatments with indicated replications ranged ( significant consistent with a 2/3,( . = 13- 13- 13- 13- Z Z Z Z et al 2, 2, 2, E E E 05, df . : 454–460 0 66 < ,malesmaybe 2010; 13-18:Ald under field values all Z population monitoring in survey traps than any P 2, -tests indicated that traps t O. sacchari E Pest Manag Sci 13-18:Ald, precise identification Z O. sacchari O. sacchari 2, 13-18:Ald. developed an HPLC technique that E 13-18:Ald present in the lure and/or Z Z 13-18:Ald as an attractant produced by 22 2, . Z 2, Z E 13-18:Ald used alone again caught the most 2, Z E et al 13-18:Ald are more difficult to interpret as the 2, 18-dienal, the field data showing that Z E 13-18:Ald, indicating that both compounds are Z 13-18:Ald caught significantly more male moths Z Z 2, Z 2, Islam 2, Z E female moths. While the retention time of the 2/3, 22 )13-octadecadienyl compounds. 13-18:Ald may be isomerized to 13-18:Ald used alone was attractive at low concentrations Z Z Z While GC-MS analysis was unable to assign stereochemistry to 2, 2, 39). Based on ancatches, examination it of was the possible to spatialwere identify distribution consistently clusters of associated of trap with trapfour locations high collection that or periods. These low ‘hot’via catches and shading over in ‘cold’ Fig. spots the 1(D). are indicated in the northern areasignificantly of fewer the field mothslure holding than on virgin each traps females of caught baited the with four sample the dates artificial ( 18:Ald is thethe most active active compound isomercatches released strongly with by suggest calling that female this moths. is Field 0.09 versus 0.46period. MTD, This respectively, compared with over 1.04trapping the MTD that in 2 held week the traps the collection of used artificial catch for lure. mass in Expressed trapsJackson as holding traps a the baited percentage artificial withthe lure, virgin field trap females averaged catch 14, in 22, in the27 4 and the northern and 31 5% half 16 July for respectively of (Fig. collection 1). dates The of 21, 24, 18:Ald and compound may be attractiveattracted to male to theZ 4% conditions. Additionally, it is possiblereleased that by the female actual moths pheromone isdata a did mixture of not aldehyde show isomers. a EAG difference between responses to the isolated 2/3, can detect the conjugatedHowever, 2,13-dienals 3,13-dienals at lack nanogram a strong quantities. derivatized chromatophore with and a must hydrazine reagent be before analysis.the As present the aim work of was to identifyin an the attractant field, that the could authors betesting chose used to to find use the electrophysiology most2/3,( active and attractant(s) field among the candidate 4 DISCUSSION Based on the GC-MS,tentatively EAG identify and field data collected, the authors O. sacchari 18:Ald matched that of 18:Ald is the more stable isomer, isas much easier opposed to to synthesize (PCC Dess–Martinchoice oxidation) as and a is sex therefore attractant the for best perceived by male moths. From a practical standpoint, and control applications. TheE first field testand caught demonstrated significantly more that of the other chemicals tested singly. Inincluded the blends, second field test, which moths, although none of the lurethe types differed others. significantly The from lack ofto statistical generally separation lower might trap be catchesfirst, partly a in situation due this expected test to period increaseRegardless, within-treatment relative variability. the to the resultsmain-effects of test for field field testcontaining test 2, 1 which were demonstrated confirmed that lures by the than those containing becomes difficult because many ofand these aldehydes isomerize are unstable duringinjection). GC analysis (even with cool on-column 2, 0, . . 1 6 The 9, 66, = www.soci.org EB Jang = over = Range F periods trapping F 3, . 0 2010 Society of Chemical Industry The average 5, df 02, . . c = 1 0 1, 58). P 13-18:Ald caught = = = Z Tukey F grouping 2, P 0, df Z 13-18:Ald caught an 5 moths per trapping . . 18, Z 6 . 13-18:Ald versus those 1 a 0 Z 2, 07) A 0.0–1.3 05) A 0.0–1.5 12) A 0.0–3.0 20) A 0.0–3.3 15) A 0.0–3.8 23) A 0.0–1.8 18) A 0.0–4.8 45)12) A A 0.0–5.0 0.0–2.5 17) A 0.0–3.7 = + ...... E 0 0 0 0 0 0 0 0 0 0 = 2, F 13-18:Ald used alone (0.22 males caught in Jackson sticky E Z ± ± ± ± ± ± ± ± ± ± SEM) P 02, Males 2, . t or binary mixtures of chemical ± pairs of traps monitored every per day trapped ( 0 Z 13-18:Ald versus those containing 0.25 ( 0.25 ( 0.37 ( 0.47 ( 0.49 ( 0.58 ( Z = 2, P E gofchemical) µ 13-18:Ald and Virgin female (A)]. During 1, 66) but not replication ( Z Opogona sacchari = 2, 13-18:OH 13-18:OH 13-18:OH 13-18:OH 13-18:OH 13-18:OH E Z Z Z Z Z Z 13-18:Ald was significant ( 8, df 2, 2, 2, 2, 2, 2, . Z Z Z Z E E E 6 2, = Z Number of 92 (95% confidence interval). F . 0 ± 01, 1, 58). The lures containing 05) (Table 3). However, the main-effects ANOVA comparing . 13-18:Ald 0.22 ( 13-18:Ald/ 13-18:Ald/ 13-18:Ald/ 13-18:Ald was significant ( 13-18:Ald/ 13-18:Ald/ 13-18:Ald/ 13-18:Ald 0.78 ( . 0 3, 66) or treatment (lure type) ( Z Z Z Z Z Z Z Z Z 13-18:Ald when used alone (0.78 MTD), which caught more 0 (1 : 1) (100 : 1) (10 : 1) 10 : 1 (100 : 1) (1 : 1) = Z Attractants were assayed using 2, 2, 2, 2, 2, 2, 2, 2, 2, = = Field test 3 (mass trapping trial using E2,Z13-18:Ald). Field test 2 (blends of E2,Z13-18:Ald and Z2,Z13-18:Ald). = Z Z Z E Z E Z E a E Virgin female (A) 0.63 ( Treatment (ratio) Virgin female (B) 0.79 ( Table 3. traps baited withlures single-componen (septa loaded with 250 3–4 days over 2–5 trappingwere periods (one carried replication). out Four replications over time.captures The main-effects in ANOVA traps comparing male containing 2, P ANOVA results). Consistent with the firstthat field test, caught the treatments thevirgin most female moths treatment were (catchingE the 0.79 two and replications 0.63than of MTD) 3 the and times the asMTD) (Table many 3). A males multiple comparison as test indicateddifferencesintrapcatchamonganyofthetreatments(Tukey’sHSD, no significant α male captures in traps containing df df containing average of 0.55 MTD, while those containing 0.30 MTD. number of male moths capturedfemales in in Jackson the traps holding mass-trapped virgin area versus the control area was www.interscience.wiley.com/journal/ps 9.0, 4.5 and 5.0[Virgin MTD female for (B), the three best-performing treatments average catch of malesranged per from day 0.79 in to 0.22 glueper (Table traps day 3). across The in treatments number traps( of was males significantly caught associated with trapping row other trapping periods, traps baitedsometimescaughtnomothsatall(Table with these same 2).Variationovertrapping attractants replications (comprising 2–5 trapping periods each) waslow relatively by comparison (SEMthat values) the (Table specific 2). locationvariable There affecting of was trap individual evidence catch. trapspairs The mean holding was difference the an between same important trap attractant was period

458 459 larvae in rotting males (5–10-fold O. sacchari O. sacchari www.interscience.wiley.com/journal/ps 13-18:Ald. While the mating disruption trial was Z 2, e key to the symbols used is shown in Fig. 1D. The shaded E 13-18:Ald, this technology is expected to be useful on Z 2, E collected in Jackson traps holding either a virgin female moth or a In the mass trapping trial and in other field studies described www.soci.org in this manuscript, it wascaught observed that relatively certain larger traps numbers consistently of with 10 mg of unsuccessful, mass trapping was a partial success,successful although as not the as trial reported in this50 manuscript. mg The fact dose that the rate provided10 mg better dose mass trapping rate results usingthat the than an failure the identical to shut placement downremedied of attraction by to traps increasing virgin the suggests females release could rate be of the attractant. ACKNOWLEDGEMENTS The authors wouldand Esther like Schneider to for thank their Janice assistance Nagata, in conducting John field Ross more) than adjacent trapspatterns sometimes that persisted over were a less periodother trap of than locations weeks, began to 7 after catch m relatively which large numbers.on away. Based these Such data, itwere was being hypothesized caught that in mosttraps traps of in emerged which the as they males adults wereused that very caught. close in In to the earlier the abandoned papaya studies, field counts of papaya trunks adjacent tomade high- during and timed low-trap-catch searches.the areas given The were hypothesis, results i.e. weretraps larval that consistent counts regularly with were caughtshown). higher greater numbers adjacent This of to suggests mothsare that (data efficiently males not caught have soonrelatively a after low sensitivity emergence low and as short dispersal active-space adultsthis response rate or range lure. to or have If a matingusing disruption or mass trappingthe can be scale achieved of individualcontrol might farms, be as relatively slow. reinvasion following successful h was either consistently high or consistently low over the 2 week sampling gwas Opogona sacchari µ 13-18:Ald. Z 2010 Society of Chemical Industry 13-18:Ald, it 2, Z c 13-18:Ald lure, E 2, Z E 2, E 13-18:Ald. Traps of the same type were spaced 6.3 m apart. Th Z 2, E : 454–460 Half-life increased dramatically as the number 66 26 Opogona sacchari 2010; Graphical and spatial representation for the number of Based on the cost for the synthesis of the Attractant for the cost of the chemical$US in 0.10. These each lures septum provided dosedmore effective with attraction than 250 in the 2 weeks fieldusing for under septa east supplied by Hawaiithat a conditions. this cooperator 2 Recent week (data period tests not canappropriate be shown) chemical extended suggest to stabilizers at are least addedUsing 8 to weeks the when the synthetic pheromone tolower-cost attract option compared males with is using a virginvirgin females. simpler Producing and females isinsects time are intensive difficult and tobeing handle expensive. transferred because into Further, they cages these direct escape and sunlight easily within are when a easily closedtrapping killed container. trial The if results indicate exposed of thepopulation to the control of potential mass banana moth. of Although it proved using impossible completely to this shut down technique trap catch in for traps holdingthe virgin traps females, holding the artificial lure, interspersed with thosevirgin holding females, caught moreauthors than 10 plan times to asdispenser, repeat many which moths. this may The trial releasehalf-life of using the various moth a attractant pheromones loadedsepta more PVC (like onto the quickly. natural or ones rubber used The in polyethylene theand present McDonough. trial) was studied by Butler is reasonable to surmise thatwas the not bulk of released thisperiod. from 18-carbon Prior material septa to during themanuscript, the mass the present trapping authors experiment 2 also week describedmass carried in trapping study out trials this in mating the disruption same and papaya field using septa loaded of carbon atoms incarbon the atoms molecule had increased. half-lives Alcoholsthose with having ranging 14 carbons 12 had from half-lives ranging 14Alcohols from 117 having to to 202 16–18 45 days. days, carbonsAlthough had while half-lives Butler of and 399–1117 McDonough days. did not test Pest Manag Sci septum dosed with 50 mg of areas enclosing groups of trapsperiod. indicate trap locations where the trap catc Figure 1. , , . et al et al JAm et al J Appl JChem JChem :189–195 :417–422 : 454–460 Nemapogon 33 17 ZNaturforsch 66 JPesticSci Chamaesphecia 2010; and Entomol Experiment ). ae) in sapindaceous fruits Biosci, Biotechnol Biochem ˆ uda V and Ivinskis P, Sex . sing sex attractants for tineid MassSpectroscopy 2,13- and 3,13-octadecadienals d tineid species (Lepidoptera, of 3,13- and 2,13-octadecadienyl Biochem Systemat Ecol Pest Manag Sci :153–159 (2001). :1848–1854 (2000). decadienyl acetate. 11 93 C. empiformis Nokona pernix ´ aki G and Schwarz M, 2,13- and 3,13- :1763–1773 (2007). . SAS Institute, Cary, NC (2008). Synanthedon vespiformis ´ aki G and Sauter W, Sex attractants for male (and/or 33 spp. (Lepidoptera: Tortricid :7277–7287 (1991). ˆ Chemoecology uraitis R, Miatleuski J, B J Econ Entomol :342–355 (1992). :203–205 (1983). 113 ´ oth M, Szir : (3Z,13Z)-3,13-octa 113 ¨ 34 ocs G, Szir ¨ :799–813 (1993). aeorg U, Karalius V, Rothschild GHL, Kolonistova S, Ivinskis P, JChemEcol :627–633 (1981). :1120–1125 (2001). 19 7 , C18 dienes as attractants for eighteen clearwing (Sesiidae), :163–167 (1993). :508–516 (2006). 56 et al tineid (Tineidae), and choreutid (Choreutidae) moth species. Ecol attractants for six clearwing an 18 Sesiidae and Tineidae) from Kazakhstan and Lithuania. C of the sex pheromonegranellus of the European grain moth octadecadienyl compounds compo and sesiid moths (Lepidoptera). (1989). clothes moth: 4.pheromones. Identification of male- and female-produced microlepidoptera found in field trapping tests in Hungary. Entomol clearwing moths: tenthrediniformis Applicat Cryptophlebia (1985). rates of alcohols andEcol acetates from natural rubber septa. elucidation of koiganal I and II, theclothesmoth,usingcapillaryGC/MS. sex pheromones of the webbing Dess–Martin periodinane) foror the secondary alcohols selective and oxidation aChem Soc of variety of primary related 12-I-5 species. from Hawaii. Synthesis and characterization of for the identification of the sex pheromonemoth. secreted by a clearwing Synthesis and characterization compounds for identificationby a of clearwing the70 moth, sex pheromone secreted ¨ ocs G, T ˆ ´ uda V, M oth M, Sz SAS System for Windows 13 B 14 Karalius V, Moz 16 Sz 15 Ohshima M, Kuwahara Y, Matsuyama S and Suzuki T, Identification 17 Takacs S, Gries G and Gries R, Communication ecology18 of T webbing 19 Voerman S, Audemard H and Priesner E, Sex attractants for 20 Follett PA and Lower RA, Irradiation to ensure quarantine security for 26 Butler LI and McDonough LM, sex pheromones: evaporation 25 Yamaoka R, Shiraishi Y, Ueno T, Kuwahara Y and Fukami H, Structure 21 Dess DB and Martin JC, A useful 12-I-5 triacetoxyperiodinane (the 23 Naka H, Nakazawa T, Sugie M, Yamamoto M, Horie Y, Wakasugi R, 22 Islam MA, Yamamoto M, Sugie M, Naka H, Tabata J, Arita Y, 24 Proc Proc .CAB (Bojer, www.soci.org EB Jang :33–36 )-3,13- Z Opogona :189–192 :457–460 , 35 Z 22 103 2010 Society of Chemical Industry c :41–43 (1984). Opogona sacchari 51 Sanninoidae exitiosa , a tortricid pest of (Lepidoptera: Tineidae) :123–132 (1982). Opogona sacchari 39 H NMR and Douglas Light 1 (Bojer), USDA APHIS (2009). :47–49 (1983). Quarantine Pests for Europe 50 :302–314 (2001). (Bojer) (Lepidoptera Hieroxestidae). ,in 94 :593–618 (1990). , pp. 1–4 (2005). Entomol Experiment Applicat Proc Florida State Hort Soc :97–98 (2005). Proc Hawaiian Entomol Soc 92 Ereunetis simulans 37 Arquiv Inst Biol Sao Paulo (Bojer), and its introduction into Florida. Cryptophlebia batrachopa Opogona sacchari ˜ na JE, Biology and morphology of the banana moth, J Econ Entomol Opogona sacchari Opogona sacchari CTAHR CES IP-21 . 1 – ensaios preliminares sobre determinacao dos Arquiv Inst Biol Sao Paulo ˜ International/EPPO, University Press, Cambridge, UK,(1992). pp. 218–221 moth on foliage plants. (1990). Opogona sacchari Hawaiian Entomol Soc (Lepidoptera: Tineidae), a new pest of pineapple in Hawai’i. Entomol Soc Washington coffee, in Boll Lab Entomol Agraria Filippo Silvestri sessuale di against Mediterranean fruitTephritidae). fly and oriental fruit fly (Diptera: Dodecenyl acetate:pheromone a of majormacadamia component in(1984). Malawi. of the female sex to the sex(Lepidoptera: Sesiidae). attractant of peachtree borer, sacchari componentes ativos. 1856). feromonio sexual de traca da banana, (1977). na JE, Duncan R and Torres V, Dynamics and control of the banana 2Pe 3 Davis DR and Pe 1 Data sheet on 4 Vorsino AE, Taniguchi GY and Wright MG, 5 Nelson S, Easton-Smith V and Wright MG, Banana moth as a pest of 8 Rotundo G and Tremblay E, Osservazioni preliminari sul feromone 6 Regulated pest list: 7 Armstrong JW, Quarantine security of bananas at harvest maturity 9 Ioneda T, Fell D and Giannotti O, Estudos preliminares sobre o 12 Brennan BM, Attraction of 11 Hall DR, Beevor PS, Cork A, Nesbitt BF and La Croix EAS, (Z)-8- 10 Ioneda T, Fell D and Giannotti O, Feromonio sexual de www.interscience.wiley.com/journal/ps REFERENCES experiments, electrophysiological experiments and moth rearing, Randy Akao (WH Shipman Limited)Nurseries, and Inc.) Patrick for McGrath permission (Hawaii Dracaena to fields, carry Scentry outoctadecadienol, Biologicals the tests Shenandoah in Valley for NMR papaya Consortium samplesat housed and James Madison of University for ( use of and Aijun Zhangproject for was comments funded on byAgriculture an (Agreement a No. earlier grant 58-5320-7-423). manuscript. from This the Hawaii Department of

460