1397 014 .Two a,c These nets are 11 – Cucumber mosaic 9 Bemisia tabaci and L. (Lepidoptera: Plutellidae) and María Plaza, a,c b,c and in brassica crops. 8 Aphis gossypii , 7 Kaltenbach (Hemiptera: Aphididae), but not Plutella xylostella (Fabricius) (Lepidoptera: Crambidae), or the a parasitoid commonly used for biological control was compatible with LLITNs. Myzus persicae Lipaphis erysimi Hellula undalis Elisa Viñuela Correspondence to: Beatriz Dáder, Departamento detuto Protección de Vegetal, Ciencias Insti- Agrarias (ICA-CSIC), C/Serrano 115 dpdo,E-mail: 28006 [email protected] Madrid, Spain. Departamento de Protección Vegetal, Instituto de Ciencias AgrariasMadrid, (ICA-CSIC), Spain Protección de Cultivos,(UPM), E.T.S.I. Madrid, Spain Agrónomos, Technical University ofAssociated Madrid unit IVAS CSIC-UPM, Madrid, Spain Field experiments using LLITNs have demonstrated promising ∗ c a b applied to the net surfaceincorporationintheprocessofmakingtheyarnsinthefactory.In by immersion or spraying,the but latter also case, by the nets arenets called (LLITN), long-lasting insecticide-treated and theunder insecticide field conditions. may persist more than 3 years results against agricultural pests such as mitesresulting in in African higher eggplant, yields, cost effective in cabbage production. LLITNsbarrier to serve control a as wide an range effective of lepidopterandiamondback pests, moth including the and aphid b,c A. colemani 1 Aránzazu Moreno, Aphidius colemani, a,c This strategy was 5 For this reason, new .Theefficacyof 4 , Fermín Amor, 3 The insecticide may be 6 a,c The selection of an appropriate 2 Saioa Legarrea, a,c a,c* : 1397–1406 www.soci.org © 2014 Society of Chemical Industry 71 2015; Treated nets; aphids; whiteflies; parasitoids; virus control; SADIE Cucurbit aphid-borne yellows virus

and

Insecticide-treated nets were developed long ago as bednets in Keywords: nets were selected for field tests under avirus high insect infestation pressure in the presence ofof plants aphids, infected was with studied in parallel field experiments. RESULTS: LLITNs produced high mortality of aphids, althoughexcluded their whiteflies under efficacy laboratory decreased conditions; over however, time they failed with in sunand the exposure. reduced field. Certain the Nets nets incidence effectively blocked of the viruses invasion of in aphids the field. The parasitoid CONCLUSION: LLITNs of appropriateadditional mesh protection against size insect can vectors of© become plant 2014 viruses a Society under of very IPM Chemical programmes. Industry valuable tool in combination with biocontrol agents for Pest Manag Sci The control of these pests generallyspraying, involves with intensive insecticide undesirable effectsand on public the health. Therefore, environment, therealternatives growers is under the an scope urgent of need IPM.an The to use excellent develop of method physical to barriers is reducedisease pest transmission access to to plants. crops and impede 1Vegetable INTRODUCTION crops suffer from economicallyand damaging insect-transmitted insect virus pests pathogens. Integrated pestment manage- (IPM) programmes entailof an chemical interdisciplinary and combination biological measures to manage pest damage. BACKGROUND: Long-lasting insecticide-treated nets (LLITNs) constitute a novel alternativetactics that to combines prevent physical insect and access chemical and the spread ofon insect-transmitted a plant viruses slow-release in protected insecticide-treated enclosures. This net approacha with is wide based large range hole of sizes LLITNs that was allow tested improved under ventilation laboratory of conditions greenhouses. against The efficacy of Abstract nets Beatriz Dáder, Michele Carmo-Sousa, protected crops by novel pyrethroid-treated (wileyonlinelibrary.com) DOI 10.1002/ps.3942 Control of insect vectors and plant viruses in Research Article Received: 1 August 2014 Revised: 27 October 2014 Accepted article published: 18 November 2014 Published online in Wiley Online Library: 11 December 2 strategies together with physicalto barriers prevent need damage to due to be smalldence developed of insect plant pests pathogens. and reduce the inci- public health to give protection against malaria. approved for use with pyrethroids, compounds that exhibitknockdown a rapid effect and highwithout mutagenic insecticidal or teratogenic potency effects. at low dosage insect screen depends on severalinsect, factors: the the size and thoracic geometry size ofinterlaced. of the Unfortunately, the hole effective and the barriers wayalso against threads are reduce small the insects airflowwhich frequently and increases fungal problems. the ventilation inside greenhouses, Alberto Fereres C ∘ and + et al. b net) net) 1 1 − − PBO : 1397–1406 71 compound (g kg (g kg Deltamethrin M. persicae Cucumber mosaic 2015; a 22 L. cv. ‘Primal’) (Syngenta )UV-addChemical )UV-addBifenthrin ) UV-add 2 2 2 Pest Manag Sci cv. ‘Marumba’) (Enza Zaden S.L., Cucumber aphid-borne yellows virus L.cv.‘JustRight’)(Takii&Co.Ltd,Kyoto, Cucumis melo )and ) 13 days after sowing at the one-true-leaf stage Q biotype donated by La Mayora Experimental Brassica rapa Cucumis sativus Cucumovirus Characteristics of the nets used in the glass vial experiments aphids were produced by rearing aphids at high densi- Polerovirus 11 Yellow 0.41 No – C (day:night) and 60–80% RH. Winged 11 Yellow 0.70 No – ∘ × (CMV, × Piperonyl butoxide. Addition of UV-blockers. Cucumber ( 64/11/07 Yellow421TR11-291 Yellow Yellow2 0.29TR11-290 Yellow Yellow364/11/08 0.71 0.12 Yellow No Yellow40 0.83Net 0.46 code Yellow Colour Yes No 0.60 Yellow 0.29 – Yes Yes Hole (mm 0.44 Yes 4.0 0.12 Yes – 4.0 Yes 3.8 Yes 4.0 2.1 5.0 3.4 196 Violet 2.92 No Deltamethrin 1.4C7 Yellow 0.77 No – 2.43.4C10 Yellow Yellow 0.56 0.45 No No – – 195a b Pink 2.67 No PBO Table 1. Net code Colour151210 Hole (mm C25warp Green White149 White412404 White 2.41406 0.73 White150 1.93 Light blue190 Green147 3.42 No Blue No 2.78405 3.38 No Dark blue191 Yellow 2.77206 No White148 2.62 No – No 2.65 Light – blue207 – Yellow 2.5925 Yes White 2.07 2.0 25-30 2.47 No White No 1.4 2.0 Net code 2.06 White No White Colour 1.4 2.00 No Yes 2.0 1.82 Hole (mm 2.0 No 0.66 0.35 4.0 No 1.4 2.0 No No No 4.0 4.0 4.0 1.2 2.8 A. gossypii ties on turnip ( Japan) and melon plants ( Seeds B.V., Enkhuizen,Bemisia The tabaci Netherlands) respectively.Station Whitefly (Málaga, Spain)house was facilities reared at on aand melon photoperiod 70–80% of plants 16:8 RH. h in Identityrearing (light:dark), green- was of 20–23 periodically biotype confirmed status byof of determining cytochrome the the oxidase sequence I population mitochondrial in gene. Almería, Spain) plantsvirus were inoculated with (CABYV, 23:18 was Stan- Sulzer www.soci.org B Dáder There- Bemisia 13 , 17 Cucumber 12 16 , L. (Hemiptera: © 2014 Society of Chemical Industry 15 )and Aphis gossypii Myzus persicae ). The two viruses have and so that the integration and 14 Cucumovirus Viereck (Hymenoptera: Aphidi- Polerovirus Aleyrodes proletella (CMV, Myzus persicae (CABYV, , different UV-blocking additives and various 2 Finally, the compatibility of LLITNs with the Glover (Hemiptera: Aphididae). These insects 21 Aphidius colemani – 11 – 18 9 Gennadius (Hemiptera: Aleyrodidae), Cucumber mosaic virus Aphis gossypii Aphids are the most important vectors of plant viruses. Nevertheless, more accurate studies under laboratory and field The objective of our work was to test under laboratory and field and are polyphagous and cause great concernage because of by direct extracting dam- planttheir fluids, but ability more to importantly transmit because of devastating virus pathogens. inae) was tested under field conditions. conditions are necessary toaction of understand this fully novel pest the control strategynatural and mechanism its enemies of compatibility with commonly usedenclosures. in To biocontrol date, under littleand protected attention there has are no been studiesbehaviour paid or concerning performance the to of natural effects enemies. this The of bioassays issue, instudy LLITNs this were on designed to the select thenets most appropriate with LLITN among different propertiesand hole (insecticide size) compounds, against three dosages tabaci key pests in vegetable crops: understanding of control alternatives is necessary. wileyonlinelibrary.com/journal/ps 22.1 MATERIALS AND METHODS Long-lasting insecticide-treatedNets nets were made ofterns polyethylene and yarns knitted providedtrol in by SAS different the pat- (Castelnauucts companies Le Ltd Intelligent (Kibbutz Lez, Ginegar, Insect France)with Israel). The Con- insecticides and net during Ginegar yarns theLLITNs. were Plastic manufacturing pretreated Yarn process Prod- diameter to ranged23 produce from insecticide-treated 0.13 nets and toaccording ten to 0.29 the mm. untreated following criteria A controls were tested: classified totalfrom a 0.12 hole to of 3.42 size mm ranging against the cabbage whitefly Aleyrodidae). 2.2 Insect culturesA and virus clonal isolates population of insecticide compounds and dosages (Table 1). aphid parasitoid fore, it is crucialof to potentially interfere viruliferous vectors with inside the protectedstudied crops. two immigration major We aphid-transmitted and have plant viruses dispersal affecting cucur- bits: aphid-borneyellowsvirus different transmission modes. CMV is transmitted in anon-persistent stylet-borne, manner duringwhile brief CABYV probes is on avirus epidermal circulative, that requires cells, non-propagative long phloem-restricted feeding probes for transmission. conditions a wide rangedence of of pests LLITNs and viruses designed of horticulturaltected to crops enclosures grown (greenhouses reduce or under net-houses). pro- the The new inci- approach is based on avide slow-release relatively larger insecticide-treated mesh net sizesstandard that and thereby untreated can greater pro- airflow nets. than the Moreover, spatial the distribution effect ofwas of aphids such studied and nets using aphid-transmittedmethodology. viruses on spatial analysis by distance indices (SADIE) dard insecticide applicationsinsect resistance have to many led substances, to the development of started from a singleand virginoparous Almería apterae (Spain) respectively. collected Both intained aphid in colonies Madrid a were growth chamber main- at a photoperiod of 16:8 h (light:dark),

1398 1399 ′′ C ∘ 9), 05 = ′ mor- n 29 hole size; B. tabaci ∘ Solanum 2 Therefore, Insecticide N, 3 24 radiation dur- L. cv. ‘Ashley’) 25 ′′ with a tomato 1 and 20 M. persicae − 58 6), using a turnip ′ = day 18 n radiation during win- 2 ∘ were released in the − 1 C. sativus − B. tabaci C maximum temperature, 9) and the persistence of A. gossypii ∘ 3), net colour (white versus day = 2 = n − n hole size; net 2, 0.60 mm B. tabaci 2 C maximum temperature, 1.8 ∘ wileyonlinelibrary.com/journal/ps and 50 using a cucumber leaf ( 4) in the nets. Daily average climatic data were = n L. cv. ‘Marmande’) (Semillas Fito S.A., Barcelona, C mean temperature, 23.4 3) was tested using a tomato leaf as target ( 4). These nets were also tested in glass tube tests M. persicae ∘ 6) and chemical compounds [deltamethrin, piperonyl = = = n n ( n A. gossypii C mean temperature, 11.7 C minimum temperature and 21.1 MJ m hole size; net TR11-290, 0.46 mm ∘ ∘ 2 In parallel experiments, net samples were placed in trays and The influence of deltamethrin concentration on Bifenthrin was also tested because it has been reported to have bottom chamber for the deltamethrin trials. exposed for 1 month to fieldtal Farm conditions (Arganda del at Rey, La Madrid, Spain) Poveda (40 Experimen- lycopersicum Spain). Ten W) during winter and springdelthametrin seasons ( to assess the persistence of 6.4 tality was tested with differentpresence insecticide of UV-blocking concentrations additives ( ( yellow) ( butoxide (PBO) and a combination of both] ( leaf as target. TheB. tabaci efficacy of deltamethrin-treated nets against ter, and 15.8 a similar experimental procedure wasent applied types to of compare bifenthrin-treated differ- netbifenthrin in ( nets exposed to field conditionsautumn 2 months season during the at theMadrid) field ( experimental site (Arganda del Rey, against (Rocalba S.A., Gerona, Spain) andleaf against (cv. ‘Marmande’) as targets. Twenty individuals were tested in each vial for the bifenthrin assays. 2.4 Determinationof of LLITNs the insecticide concentration Insecticide analysis was donemethod using 454/LN/M/3.2 for an alpha-cypermethrin by adaptation gasraphy of chromatog- the with CIPAC flame ionisation detection (GC-FID). minimum temperature and 6.5 MJ m ing spring. a longer persistence and stability than deltamethrin. was determined by extraction(200 mg) in in a xylene conical flask. (25 The mL)and flask of was heated connected net to to a samples condenser refluxpletely for dissolved. The 60 min solution until was cooled the to net room sample temperature, was com- 8.9 C ∘ feeding on the target after crossing a range of bifenthrin-treated nets with different hole hole size; net 3, 0.44 mm 2 Bemisia tabaci 4cmindiameter)separatedby × C). ∘ Trials were conducted in the laboratory C (day:night) and 90% RH. The CABYV iso- ∘ 23 : 1397–1406 © 2014 Society of Chemical Industry hole size; net 64/11/08, 0.29 mm 2 71 adult aphids were put on CABYV-infected source hole size). 2015; 2 (a) Schematic diagram of the laboratory experimental set-up. Half of the drawing is shown without the black cardboard covering the tube, to A. gossypii net 1, 0.83 mm Pest Manag Sci Figure 1. show the inner structure. (b) Meansizes percentage (net of 40, 0.12 mm the test net. Anno untreated net net were of used thereleased as same in controls mesh the in and bottom every vials chamber,host and experiment. plant with a was Insects leaf placed were in ofsurrounded the with a black top cardboard preferred except chamber at aphid the as top, which target.ered was cov- The with vial a was thin clothtion that into allowed the ventilation structure. and Both the lightthe target penetra- top leaf and were light the coming stimuli from the LLITN. to Insects induce located either insects feeding on toalive the climb target in leaf, up the dead release or and chamber cross wereand assessed whitefly 6 release and respectively. 24 Mortality h valuesusing after were aphid Abbott’s corrected formula. 2.3 Laboratory assays Experiments were conducted atences the (ICA) of Institute the Spanish of National Research Agricultural Council (CSIC,Spain). Madrid, Sci- A novel experimental set-up wasto designed to go force insects through theness insecticide-treated of the net net so killingassays the that were insects conducted the could using effective- be an evaluated experimentaltwo (Fig. tube glass 1a). composed cylinders All of (12 cm long (day:night) and 60% RH. plants during 48 h, and nymphs producedallowed during to this period feed were for oneperiod extra (AAP) day, reaching of an 3 acquisitionCABYV-infected days. access plants After were the transferred AAP, to 25plant each for nymphs an healthy inoculation grown access receptor period on (IAP) of the 3were days. manually Then, nymphs removed. CABYV-infected plants werein maintained a chamber at a photoperiod of 16:8 h (light:dark), 20:16 late was kindly provided by Dr H Lecoq (INRA,from France) and zucchini obtained squash in 2003lated in to Montfavet cucumber (France). plants It (cv. ‘Marumba’) was bysion. inocu- aphid serial transmis- Pest and plant virus control by long-lasting insecticide-treated netsand used 5 weeksmechanically post-inoculation inoculated as with viral CMV sources.melon Plants isolate crop were in M6, 1996 obtained inDr from E Tarragona Moriones (Spain) (EELM-CSIC, a Spain). and CMV-infected plants kindly weretained main- provided at ICA-CSIC by in an insect-proof16:8 chamber h at (light:dark), a 25:20 photoperiod of www.soci.org at room temperature (22–24 2 ∗ 1, 1. < > et al. ,and Ia Ia vi -test or 1) or 2 H + x : 1397–1406 71 , the minimum ,ln( 0.05) using IBM 2 D x ≤ P wastestedinasep- 2015; In our study, a value ; Agrobio, La Mojon- 0.5, C temperature, 70.3% 21 ® ∘ + -test ( x t √ denotes the pattern of the Ia A. colemani 20 1 and a regular sample if . Pest Manag Sci vj = When the data did not follow the , the minimum value of the total Ia 29 C 0.05) was performed. Insect occupancy ≤ , the positive patch cluster index, P Ia radiation and 2.3 mm rainfall. 1 were released on each of 11 marked plants , and contoured as well. − X -test ( U day 2 − The spatial pattern of data was described by the 19 , if needed to reduce heteroscedasticity and achieve nor- A. gossypii 18 x Moreover, the degree of local association between aphid √ 28 at 2 weeks after aphid infestation. Insect sampling was per- 2 − 2.6 EffectAphidius of colemani bifenthrin-treated nets on the aphid parasitoid The impact of the TR11-290 net on arate and independent set of threesions tunnels with as the same those dimen- previouslyExperimental Farm during described the autumn and season. We locatedilar followed a split-plot on sim- design La with Poveda divided three replicates, into each two net-house equal being a experimental section plots. of Each either bifenthrin-treated plot or contained untreated net (0.46 mm presence and virus incidencespatial association, was calculated with the index of the negative gap cluster index, index of aggregation, formed weekly for 6 weeks byall scouting plants their and absence/presence in by countingDaily their average number climatic conditions in were the 15.7 11RH, marked 12.9 MJ plants. m inside each experimental plot. As opposed to the fieldwhere experiments aphids and whitefliesthe were net-house, released the parasitoids on (APHIcontrol theera, outer Spain) side were of releasedplatform placed inside in the the net-housem centre of and each hung plot on at a a rate of 10 adults hole size) on each side ofplants, the distributed tunnel. in Each seven rows. plot At heldadults the 42 two-true-leaf of cucumber stage, three 2.7 Spatial analysis The spatial distribution ofusing aphids the and SADIE virus methodologyThis applied spread analysis is to was based on the studied the distance data tovalue regularity, of year of 2011. thethat total the distance population isthe that distributed distance individuals as to have regularlydistance crowding, to as that move possible, individuals and so possible. must move to be as aggregated as arcsin Mann–Whitney ANOVA assumptions, a non-parametric Kruskal–Wallis mality. Differences among nets in the percentageinsects of feeding mortality and on the targetthe insect leaf density in in field experiments the were assessed laboratory byric a assays one-way paramet- ANOVA and test in followed by pairwisesignificant comparison differences for (LSD) least or a Student a spatially random sample if population, which by convention is an aggregated sample if of 1 was assigned to plantsand infested by a aphids value or of infected 0 bythe virus to 42 cucumber uninfested plants or inside non-infected each plot. plants for each of 2.8 Statistical analysis Data were previously transformed with Statistics SPSS 21.0 software. For units within patchesanother, of the patch relatively cluster large indexgap counts would cluster close be index to large. tends Conversely, tocounts one the be close large to in units onelocation within and another. extent gaps of Both of cluster small indices incontoured the with visually data, a so indicate contouring their modelling values the Surfer, program could which be such allows as the Golden graphicalgaps. representation of patches and . 1 1 − m, − μ radi- 6.5 m 1 www.soci.org B Dáder − × and 500 day 2 − © 2014 Society of Chemical Industry C temperature, [0 (0 aphids), 1 ∘ hole size) either 26 2 A. gossypii Net samples were also var. ‘Marumba’) plants, 27 m) and analysed by GC-FID μ or untreated. In 2013, accord- 1 − C. sativus radiation and 1.5 mm rainfall during per experimental plot were released. L and a flow rate of 300 mL min 1 μ − , either treated with 2.1 g bifenthrin kg 2 day 2 − B. tabaci C temperature, 69.32% RH, 13.3 MJ m ∘ crossing the net. On the outer sides of the net-houses, and 400 per plot were released on the virus-infected cucumber 2.6 m high), 5 m apart and with the same E–W orientation, 0.25 mm) (Agilent Technologies Inc., Santa Clara, CA) with 4.3 m wide) was placed on each side of the tunnels, replac- × × × 150 aphids)]. Leaf samples from each cucumber plant under B. tabaci > wileyonlinelibrary.com/journal/ps an additional row of six cucumbertwo plants infected with (two CABYV infected and with two CMV, to non-infected) was provide transplanted inoculum sources2011, at 2 both days sides of after the transplant, net-house. 240 In winged net or untreated, with theing aim of decreasing the numbers of liv- filtered through a nylon filter (0.2 with a systemchromatograph, comprising an Agilent an Technologies 7693A Agilent autosamplera and capillary Technologies fused silica 7890A column (5% phenyl gas methyl siloxane 0.25 an injection volume of 1 30 m 2011, and 16.1 ation and 1.0 mm rainfall during 2013. (1–4 aphids), 2 (5–19 aphids), 35( (20–49 aphids), 4 (50–149 aphids), treated with 3.8 g bifenthrin kg collected at different time intervals during each field experimentmeasure to the remaining insecticide concentration aftersure. field Daily expo- average climatic conditions were 15.2 wide soil properties and irrigation regimes,was were used. divided Each net-house into twoa vertical equal net experimental followingEvery plots a plot separated split-plot held with design 42 withdistributed three cucumber in replicates. ( sevenlong rows. An experimentaling net the section standard transparent (3.5entire m polyester net-house. netting During that 2011, covered thetest net-house the net contained of a equal yellow mesh (net TR11-290, 0.46 mm 2.5 Efficacyand of whiteflies LLITNs in field conditions against aphids Field trials were designedtion to dynamics test and the plant incidencenets viruses of that in pest provided popula- natural thecould be conditions. best produced on The a results large two scale inments were at tested the La in Poveda two Experimental laboratory Farm field during experi- autumn trials of2013. 2011 and and Three identical tunnel-type net-houses (8 m long Insecticide content wascalibration. calculated with an external standard ing to the results oflaboratory the trials, first the field new study netwas and reduced after 64/11/08 to a 0.29 was mm second tested. set The of hole size B. tabaci plants that were transplanted onAphids the were outer directly released side on of the leaf the surfacement net-house. to and improve settle- viral acquisition. Whiteflies werelevel released at along the the canopy virus-infected sourceA. plants. gossypii In 2013, 360Aphids winged and whiteflies crossing the nets were assessedtheir by absence/presence counting in the cucumberimental plants plots. inside Furthermore, the exper- aphidber density plants in inside 11 each markedplants experimental cucum- on plot the and outer in sides thelowing was monitored virus scale weekly source previously by used using in the fol- similar studies: experimental plots were collectedand virus 9 infection weeks was after confirmedenzyme-linked by insect immunosorbent double-antibody release, assay sandwich (DAS-ELISA)commercial using antibodies specific against CMVCABYV (Agdia (Sediag Inc., S.A.S., Elkhart, Longvic, IN) France). and 61.2% RH, 12.3 MJ m

1400 1401 0.01 0.01 0.01 0.01 0.01 0.01 < < < < < < 4.6% in ± presence on 79 0.03 59 50 66 0.25 06 30 12 85 ...... FP (Table 2). The numbers A. gossypii 2 0.078). = wileyonlinelibrary.com/journal/ps P 6a 3b 2c 9c 7b 4ab5 9a34 7 4b 9 2b 7a 0b 2a21 21 1b 0b 1b 1a 1b 7b 8b 0b 4a17 9ab 0a22 0c14 0a17 ...... SE Insects on target (%) 0.4% in net 206, increasing up to 17.1 03 31 74 81 35 47 05 011 47 01 07 613 00 27 07 11 00 11 310 11 00 61 65 58 21 79 00 46 ...... X ± in the target leaf, and even after winter exposure for mortality was found to be low in this experiment, showing PBO 6 All the bifenthrin LLITNs tested reduced + net 25. However, significant differences in the percentage offlies white- reaching the targetnets were 1, 2 found and 3 between and bifenthrin-treated untreated netsthe 1.4, treated 2.4 nets tested and during 3.4 this (Table first 3). laboratory Among survey, net 3 had M. persicae net 404. Net 406 was not affectedover, by sun no exposure significant (Table 2). differences More- innet mortality colours white were and found yellow ( between the target. The five treated nets statisticallyof differed in aphids the reaching number the plantnets target, had and a the hole two size most of promising 0.71 and 0.44 mm of aphids reaching the leaf significantly differedfield among exposure. The periods unexposed of and one-month-field-exposed nets demonstrated relatively good performance, with similar valuesaphids for reaching the target. However,for when 2 the months, net the percentage was of exposed values aphids similar in to the those leaf of increased vials up with to untreated nets (Table 2). 3.2 Efficacy ofOver LLITNs against 80% whiteflies in of laboratory trials thewere whiteflies alive tested in onB. deltamethrin-treated tabaci the nets target chambera minimum 24 of h 0.1 after release (Table 3). 0.05) ≤ P Spring exposure 20 Winter exposure 1 Spring exposure 33 Winter exposure 34 Spring exposure 55 Winter exposure 25 goodness-of-fit 11 – 38 2 × 04 0 30.44 06 5 20.60 196 Deltamethrin 190 Deltamethrin 6 195 PBO 46 412 2 (unexposed) 7 404 No 0 0 b 405 Yes 7 406 Yes 14 406 1.4 (unexposed) 14 148 4 1 150 2 5 𝜒 TR11-290 3.1 (exposed for 2 months) 38 TR11-290 3.4 (exposed for 1 month) 1 TR11-291 0.71 0 TR11-290 3.8 (unexposed) 1 TR11-290 0.46 1 net) C10 net) 404 1.4 (unexposed) 0 1 1 PBO nets. PBO-treated net 195 − − + ) 1 0.83 15 on the leaf when compared with 2 net) 151 – 46 1 − Almost half the aphids reached the tar- . 30 : 1397–1406 © 2014 Society of Chemical Industry M. persicae 71 Persistence (g kg Compound 151 – 64 Persistence (g kg UV-additives 151 – 64 and thereby reduced the chances that insects 2015; Percentage of aphids feeding on the leaf target (%) in laboratory trials, showing the parameters evaluated and values, the nets used in each 0.05) to check whether the observed frequency distri- ≤ M. persicae P Bifenthrin Hole size (mm Deltamethrin Dose (g kg experiment and the statistics according to a one-way ANOVA test ( Insecticide Parameter evaluated Net code Parameter value Table 2. Pest Manag Sci 3.1 Efficacy ofAll LLITNs against LLITNs aphids impregnated in laboratory with trials ity pyrethroids produced in high mortal- would reach the target 3RESULTS rate and virus incidence were compared by a get chamber in untreated netdeltamethrin 151 tubes, and with differed fewer significantly insects from high reaching concentration the target (Table under 2). The404 addition did of not UV-blockers make any toof aphids difference net reaching with the respect target, to althoughin it the was untreated percentage significantly net higher (Table 151 2). No compared synergistic effect of with the insecticidesPBO LLITNs deltamethrin and was 404, detected. 405 Inhigher and contrast, number PBO 406 of alone caused a significantly Pest and plant virus control by long-lasting insecticide-treated nets www.soci.org bution was related to theStatview 4.01 expected software. frequency distribution using test ( deltamethrin and deltamethrin had lower values than untreated netdeltamethrin-treated 151 nets (Table decreased over 2). time The with efficacy sunin of two exposure of the three nets tested, 404the and spring-exposed 412 (Table nets 2). significantly In increased both cases, the percentage of et al. 0.01 0.01 < < : 1397–1406 29 80 46 0.91 71 . . . FP 0.01) and CABYV 2 contingency table < × 2015; P alate density (scale 0–5) 8.26; = 2 𝜒 Pest Manag Sci Aphis gossypii 8a 9a 0a19 8b 3b 8c 5b 8 4 7 1 0 3 8 7 4 4 2a17 70 2a 7a ...... SE 0.05) and (b) a chi-square 2 0.05). < Insects on target (%) < P P 71 15 81 20 95 54 23 44 12 23 95 43 83 73 74 25 45 92 31 14 74 ...... X SE values: (a) 0.01) was significantly higher in untreated plots ± < P Mean 8.07; = 2 𝜒 goodness-of-fit test ( ( than in plots coveredinfections by with bifenthrin-treated both nets viruses (Fig. were 2b). also Mixed significantly lower in plots Furthermore, the incidence of CMV ( Figure 2. and (b) CMVbifenthrin-treated and and untreated CABYV nets during virus theAsterisks field transmission indicate experiment statistical in (%) differences 2011. according inside totest (a) followed the by a DMS one-way ( plots ANOVA under 0.05) ≤ P 2 in 2.3; 2 0.01). www.soci.org B Dáder =− < Z P were not 2 8.52; © 2014 Society of Chemical Industry 443.0; = 2.46% whiteflies 11 – 94 = 2 ± × 20.6059 30.4441 10.8371 U 𝜒 25 0.66 72 2.43.4 0.56 0.45 83 83 149150190191148206 3.42207 2.65 2.62 2.07 2.00 2.06 85 1.82 84 80 81 83 83 86 147 2.59 82 25-30 0.35 78 TR11-290 3.1 (exposed for 2 months) 92 TR11-290 3.4 (exposed for 1 month) 84 TR11-290 3.8 (unexposed) 39 0.07) and 20 days onwards = P ) 1.4 0.77 90 ) 151 2.41 82 2 2 2.7; =− hole size) and provided a physical con- 0.03) respectively. Plots protected by Z 2 = reaching the target in a wide range of nets P was significantly lower in plots protected by 390.5; Persistence (g kg net) C10 2.2; = U B. tabaci =− Z Percentage of whiteflies feeding on the leaf target (%) in laboratory trials, showing the parameters evaluated and values, the nets used in A. gossypii 448.0; 0.02) (Fig. 2a). Numbers of apterae and nymphs were also = After these trials, and according to the results obtained during Bifenthrin Hole size (mm Deltamethrin Hole size (mm Insecticide Parameter evaluated Net code Parameter value Table 3. each experiment and the statistics according to a one-way ANOVA test ( = U bifenthrin-treated nets had a significantly lowerrate aphid from occupancy the second sampling date onwards ( ( 3.3 Efficacy ofThe LLITNs nets in that field gave conditions best results inand laboratory could conditions be every produced year onditions.Thenetsusedhadaholesizeof0.46and0.29mm a large scale were tested in field con- reached the target, and was therefore selected forstudy the (Fig. second 1b). field wileyonlinelibrary.com/journal/ps pore size gave promising results, as only 6.77 the lowest value (Table 3). Sunnet used exposure in of the the first field bifenthrin-treated experiment causedin a efficiency significant reduction against whiteflies under laboratorymonth conditions onwards from (Table 1 3). the first field experiment,tory we assays undertook testing a two second new64/11/08 set LLITNs and with 40) of a (Table labora- 1). smaller Figurehole hole 1b size size shows and (nets the relation between tested during several years. Pore sizes above 0.44 mm trol, as no whiteflies were foundwith on standard the net leaf 42. in the However, untreated LLITN vials 64/11/08 with a 0.29 mm 2011 and 2013 respectively.of In alate the first field study, the density the bifenthrin-treated net than in thosenets covered from by 27 days the after untreated aphid release onwards ( P lower in plots protected withdays the onwards bifenthrin-treated ( nets from 34 sufficient to prevent living whiteflieswas too feeding dense on (0.12 the mm leaf. Net 40

1402 1403 1.5. 1.31; =− = v Ia 0.00) (Fig. 3). = P and CMV in the third 1.78; = Ia , the positive patch cluster index, 0.00) (Fig. 4). In contrast, only Ia A. gossypii = 0.97), whereas it was aggregated P = wileyonlinelibrary.com/journal/ps 0.03) (Fig. 3). For CABYV, the con- P = 1.5 and blue lines are contours of 1.57; P = = v 0.81; Ia = 0.35; Ia = in plots with the bifenthrin-treated and untreated X A.gossypii 1.49 (clustering slightly exceeding expectation) and large filled spots 0.04; net-house 2: ± Owing to higher virus transmission, year 2011 data were used to = a few CABYV spots were found in bifenthrin-treated plots, and The combination of aphid infestation anda virus significant association infection between showed untreated plot ( toured maps of untreated plotsrestricted showed virus-significant to patches thetered first after two rows crossing of the plants untreated that net aphids (net-house encoun- 1: study the spatial distribution ofvector viruses CMV and CABYV andnets. Spatial the patterns of aphid presence in untreated plots revealed that aphids colonised the entire area of the experimentalaggregated plot distribution, in an although this aggregationnificant was (Fig. not 3). sig- Onto the the other borders hand, next aphidexcept to dispersal for the insect was third plot, release limited in which in aphidform. distribution bifenthrin-treated was The more plots, uni- spread of CMVdistribution in followed the either control a plots, being randomsecond significantly net-house or regular ( a in regular the in the plots protected byaggregation bifenthrin-treated in nets, with the significant second net-house ( P 1to ± , circled by coloured lines are statistically significant. The letter N and arrow and CMV-infected plants, and contoured map of the association between X 8.73; 0.51). = < 2 ), which P 2 𝜒 Aphis gossypii 0.51; 1.4%. However, = net. ± 2 1 𝜒 − 0.01); just one of the < , during the field experiment in 2011. Spots indicate individual test plants. Small filled spots represent P 7.14; = A. gossypii , and the index of spatial association, 2 vj 𝜒 0.99 (clustering below expectation), unfilled spots net at the beginning of the field experiment. ± : 1397–1406 © 2014 Society of Chemical Industry 1 − 71 0.01) but similar whitefly occupancy in plots pro- 2015; < P Classed post maps of the spatial distribution of 1.5 (more than half as much as expectation). Red lines enclosing patch clusters are contours of < 9.08; 0.01). The bifenthrin concentration was lowered to 1.3 instead = 2 According to the results of this first field study, where whiteflies < , the negative gap cluster index, 1.5 or 𝜒 of the initial 2.1 g kg tected with the treated and untreated nets ( Pest Manag Sci Figure 3. CMV-infected plants and its vector, clustering indices of 0 to indicate north orientation. Pest and plant virus control by long-lasting insecticide-treated netsprotected with treated nets ( www.soci.org three plots had plants infected by both viruses, whileof the incidence mixed infection in untreatedthe net plots mesh was was not 13.3 dense enoughpersion to in control plots whiteflies, protected and with dis- bifenthrin-treated netsto was that similar under the untreated netsconcentration (data of not the shown). net The exposed bifenthrin forduring 2 autumn months decreased in from field 3.8 conditions to 3.1 g kg had promising results under laboratory conditions (Fig. 2b). Results in 2013 followed thetrol same of aphid trend occupancy from as 15 in( days after 2011, aphid release with onwards a good con- P were not effectively excluded, wein conducted another 2013 field using study a net with a smaller pore size (0.29 mm > Black lines are zero-value contours, representing boundaries betweenvi patch and gap regions. The index of aggregation, Aphids readily entered the control plots 9 daysHowever, after the insect release. bifenthrin-treated net preventedweeks. Virus aphid incidence was entry not as for high as 3 intion 2011, but significantly CABYV increased infec- inside the untreated plots ( et al. : 1397–1406 )wereused. 2 71 when exposed to 2015; access was reduced The addition of UV-absorbing M. persicae A. gossypii 11 Pest Manag Sci – 8 and net) and large hole size (0.83 mm M. persicae 1 − and at the same time pests that passed through were 4 Our results also indicate no effect of deltamethrin degradation this is theenemy. first The report size of of theventilation, the net hole impact was of big enough an to LLITN allow proper on a natural below 20% in all of the LLITNsdosage tested, (2.0 even g when kg a low insecticide likely to acquire sufficient pesticideinsect so pests that entering the number thelaboratory of greenhouse trials, living was strongly reduced. In additives to the yarnnet was because not mortality an was obstaclestandard not LLITN. to In different the contrast, when when efficacy PBO comparednet, alone of with was it the used a did to treat not the causeincrease significant efficacy mortality when in combinedexperimental aphids with design and deltamethrin. used failed Within in to the PBO this study, was the not insecticide shown synergist pyrethroids to in enhance LLITNs, efficacyranted. although Lastly, when net further colours combined yellow and investigations with whiteon had are aphid no mortality. different war- effect in one of theLLITNs nets tested exposed lost in efficacy the field. against However, the other two No differences were observed among thewere nets treated studied with when different they doses ofhalf deltamethrin. Approximately of thefeed released on aphids the leaf were wheninsecticide-treated able an net untreated to was net used reach of as the aporation the barrier. same of Therefore, target mesh the insecticide as incor- and the toagainst the aphids yarns and acted providedexclusion as additional properties of a benefits the chemical to net. the barrier physical and CABYV-infected plants, and contoured map of the association between 0.43; 0.02; 1.30; www.soci.org B Dáder = = t =− P t 0.62 versus Aphis gossypii ± 0.36; © 2014 Society of Chemical Industry 0.14; = ± X 0.94). The average 8.07) nets ( = ± , during the field experiment in 2011. Symbols and contours are as for Fig. 3. P 0.09; 0.39 versus bifenthrin-treated = A. gossypii ± t 0.98), but a significant aggregation in 0.88; = 0.03) (Fig. 4). individuals, was the same in the two treat- P ± 0.69; week 6: untreated 0.92 = ,anaphidparasitoidfrequentlyusedasa P = P 0.34; In addition, these nets produced no harmful 0.33; 4 , =− 0.43; = 3 A.gossypii X = X t A. colemani Classed post maps of the spatial distribution of 8.28) and bifenthrin-treated (43.43 0.46; 0.03 versus bifenthrin-treated 0.30 ± ± ± 0.26). 0.69; week 5: untreated 0.91 = = P numbers of mummies(28.33 were statistically similar in untreated bifenthrin-treated 0.99 0.65 P wileyonlinelibrary.com/journal/ps 4The DISCUSSION results obtained in this studysidered suggest as that LLITNs a can promising be approach con- into for protected reducing aphid crops immigration whileenvironments. allowing suitable airflow in enclosed ments on the three parasitoid0.37 sampling dates (week 4: untreated 3.4 EffectAphidius of colemani bifenthrin-treated nets on the aphid parasitoid Mummies appeared 24), weeks after and parasitoid occupancyout infestation rate the (week of crop cycle. plantsmummies The per remained parasitism rate, constant expressed through- as number of CABYV-infected plants and its vector, infection was not detected in theciation second between plot. the virus A and significant its disso- vectornet-house was 2 recorded in ( untreated Figure 4. effects on biocontrol agent in greenhouse production. To our knowledge, the border of the treated areas (net-house 1: net-house 3:

1404 1405 It is 14 .It 36 Resis- 35 B. tabaci 34 B. tabaci B. tabaci was not suffi- The size of the survival in cab- 37 B. tabaci A. proletella hole size and 5.0 g bifenthrin 2 Further experiments should con- In this sense, LLITNs could reduce 8 wileyonlinelibrary.com/journal/ps 33 var. ‘Marumba’, to Antonio J Álvarez might be the best compromise to con- 2 could explain such unsuccessful results, effectively, but our most recent finding suggests B. tabaci was able successfully to cross the bifenthrin-treated Cucumis sativus B. tabaci Recently, promising results have been obtained with the 9 net for reasonable blocking of the access of living white- B. tabaci 1 − Notwithstanding, the hole size used was sufficiently large to When properly designed, LLITNs represent a good strategy that compared with conventional production, but it hasthe also significance increased of aphidsplants as in south-eastern pests Spain. that cause direct damage to was evaluated, although nets treated withexclude bifenthrin whiteflies appeared better to than the ones treatedIt with was deltamethrin. necessary to use a 0.60 mm allow the passage ofcant small mortality insects was such observed as in whiteflies. most No nets signifi- tested when tinue to select the mostexclude appropriate mesh size and insecticidethat to a pore size of 0.29 mm trol this species. combines chemical and physical controltainable techniques to management allow and sus- reduce pesticideMoreover, LLITNs treatments could in be crops. implementedrelease in of conjunction with commercially the availableusing natural laboratory enemies. and In net-house experiments, thisthat we study, LLITNs have can confirmed reduce aphid populationsspread of as plant well viruses. as Different mesh decrease sizes the should and other be insecticides further assessed forwould the be effective interesting control to test of this strategycommercial at field greenhouses scale to again under developprogrammes. an alternative tool for IPM ACKNOWLEDGEMENTS We are gratefulPlastic to Products Intelligent Ltd Insect forthe the Control seeds of nets, SAS to andand Enza Rocío Ginegar Zaden Oliva España for S.L.Pedro Hernaiz measuring for for the supervising the hole fieldwas size experiments. supported This by of research the Spanish the Ministry nets of(grants Science AGL2010-22196-C02-01 and and Technology and to 02). AA Moreno, E Fereres Viñuela and are members of the Associate unit IVAS (CSIC-UPM): The small size of as the body0.5 mm length respectively) and are much width smaller than of those the of aphids. whitefly (0.8–0.95 and the risk of aphid infestation.importance In of biocontrol addition, in greenhouses, owing further studies to wouldnecessary be the to increasing assess the compatibility of both strategies. pyrethroid alpha-cypermethrin against whiteflies. flies through therenowned net worldwide as an under intractable pest laboratorytrol that is conditions. and difficult to one con- that develops pesticide resistance rapidly. kg tance of this whitefly to pyrethroids is wellis known, and registered this species in the Arthropod Resistance Pesticide Database. mesh and the chemical compoundthe appear production to of be effective major LLITNs, factorscontrolled and injurious in dicofol-impregnated mites in nets eggplants, even though theirsmaller size than was that of whiteflies. is likely that, when it crossed the LLITNs, ciently impregnated with thedown pesticide effect. to The suffer findings fromthat of its our knock- field experimentsnets also and showed disperseto over when a the non-treatedresults cucumber also net agree with plots was the findings used at on as a a similar physical barrier. rate Our bage field experiments using deltamethrin-impregnated nets asfence. a 4 A. net 1 − A. cole- As shown 25 , 5 Because pyrethroids 11 – 9 density was significantly under laboratory conditions. Moreover, the dispersion of net after being exposed for 2 31 1 − B. tabaci One of the drawbacks of this approach A. gossypii 3 , and 2 : 1397–1406 © 2014 Society of Chemical Industry CABYV spread and aphid density were very 71 32 32 The use of LLITNs could also help to maintain crop 2015; 11 A. gossypii was greater inside untreated plots. On the other hand, we an important biocontrol agent. This biocontrol production In our field trials, Protection against aphids would not be enough if untreated nets In our studies, the persistencejeopardised of from the 1 month insecticides onwards, appeared suggesting to thatlose efficacy nets be after partially sun exposure. However, the amount ofleft bifenthrin in the nets wasspread enough inside treated to net-houses reduce during aphid one growing dispersionmay season. be and due This to virus the lower insect densityexperiment, during the as first weeks the of first the vectorsimpregnated that and crossed the died treated before netongoing reaching were work the focuses cucumber crop. onand Our testing formulations further to UV-blocking maintainperiod additives longer the than a efficacy single crop-growing of period. these nets over a reduced in cucumberbifenthrin-treated net-houses, plants so it protected isbe likely by that extended our completely results to could closed other aphid species. scheme has reduced the number of insecticide applications when mani, sanitation in regions where vegetablebiological crops control are and produced IPM using nets programmes, as of we this have kind showna can that way in that some is circumstances compatible at with least beneficial be insects used such in as were placed as a physicalsible barrier solution to alone this in problem venthole would openings. to be A 0.34–0.40 to pos- mm. reduce the size of the months in the field,nets which against strongly reduced the efficacy of the Pest Manag Sci Our results suggest that the use of LLITNsor either with bifenthrin deltamethrin might allowLLITNs adequate placed ventilation in of theings greenhouses. sides would decrease of aphid greenhouses entry,pesticide or treatments thereby and in reducing increasing the the window safety need open- environment. of for growers and the Pest and plant virus control by long-lasting insecticide-treated netsspring conditions, possiblyradiation owing than to duringefficacy higher winter, temperature after although and winter onecontrol. The exposure of bifenthrin compared concentration them decreased with also to 3.1 the lost g kg unexposed www.soci.org produce a rapid knockdownfield effect, scale the may application of reduceby aphids, LLITNs the such at spread as ofa CMV plant and slower CABYV. viruses knockdown In transmitted compared effect particular, with bifenthrin but some has better other pyrethroids,be chemical taken an into stability aspect account that when when developing needs new to LLITNs. limited in bifenthrin-treated plots, which may indicatedispersion again a rate low of bothpopulation agents was associated under with LLITNs. CABYVplots, In infected-plants as in frequently addition, treated observed aphid in virusescirculative transmitted manner. in a persistent could be the insufficient ventilation inside the enclosed structure. in the spatialof analysis of both the virusesunder field and the experiment, mixed insecticide-treated virusand incidence infections net. CABYV spread Different significantlySADIE. inside patterns decreased CMV control spread for had plots either CMV in a were regular untreated also or plots, a found random anon-persistent using distribution result viruses, that asunder matches plots opposed the protected by to typical LLITNs. spread the of aggregation found instead of the initial 3.8 g kg found significant CABYV aggregation inplots, the borders which of untreated suggestsadjacent an plants. initial focus that led to infection in gossypii . et al. :astudy :303–314 :177–201 , Repellent :1008–1017 64 51 : 1397–1406 Bemisia tabaci 79 et al. 71 species complex :158–168 (2009). :393–424 (1990). :9–13 (2009). Coccinella septem- :1593–1602 (1999). Annu Rev Entomol 28 :1–8 (2012). 8 49 141 :684–690 (2014). 2015; Ecology JAnimEcol :106–113 (1999). 165 107 2 Barley yellow dwarf Mol Ecol Virus Res Bemisia tabaci Virus Res Ecol Lett Golden Software Inc., Golden, CO . SPSS Inc., Chicago, IL (2013). Annu Rev Phytopathol :33–45 (1994). IOBC/WPRS Bull Pest Manag Sci 72 , ed. by Ishaaya I and Horowitz AR. :133–141 (2002). 9 J Econ Entomol Annu Rev Phytopathol :475–483 (1977). . Marston Book Services Ltd, Abingdon, Oxon, 34 :265–267 (1925). Ecoscience 18 Biorational Control of Arthropod Pests: Aplication Entomol Exp Applic . JGenVirol Abacus Concept Inc., Berkeley, CA (1992). , Status and prospects of plant virus control through interfer- :431–457 (1991). et al. (1995). (1998). detecting clusters in count data. ecological data. D (2013). phylogeographical analysis of the based in mitochondrial DNA markers. ence with vector transmission. J Econ Entomol Food and AgricultureHealth Organisation Organisation, Rome, of Italy, pp. the 1–33 United (2010). Nations/World UK, p. 40 (2009). A, Dynamics of nonpersistent aphid-bornecovered with viruses UV-absorbing in nets. lettuce crops of enzyme-linked inmunosorbent assayviruses. for the detection of plant (2009). transmission by homopteran insects. the wide scale implementation ofhorticulture biological control in in Almería, Spain. greenhouse in ecological complexity. on the within-crop behaviour and distribution of punctata overview, in and Resistance Management Springer-Verlag, New York, NY, pp. 1–20 (2009). 36 effect of alphacypermethrin-treated netting against (Hemiptera: Aleyrodidae). CIPAC Handbook, Vol. M Specifications and Evaluations for Agricultural Pesticides: Bifenthrin Golden Surfer Software Version 9.0. SPSS Statistical Package Version 21.0 Statview. 19 Perry JN, Measures of spatial pattern for counts. 20 Perry JN, Winder L, Holland JM and Alston RD, Red-blue cages for 21 Perry JN and Dixon P, A new method to measure spatial association for 17 Bragard C, Caciagli P, Lemaire O, López-Moya JJ, MacFarlane S, Peters 18 Perry JN, Spatial analysis by distance indices. 22 Frolich DR, Torres-Jerez I, Bedford ID, Markham PG and Brown JK, A 23 Abbott WS, A method of computing the effectiveness of an insecticide. 25 24 26 Legarrea S, Betancourt M, Plaza M, Fraile A, García-Arenal F and Fereres 27 Clark MF and Adams AN, Characteristics of the28 microplate method 29 30 31 Fereres A and Moreno A, Behavioural aspects influencing plant virus 33 Van der Blom J, Robledo A, Torres S and Sánchez, JA, Consequences of 32 Irwin ME and Thresh JM, Epidemiology of 34 Willes JA and Jepson PC, Sub-lethal effects of deltamethrin residues 35 Horowitz AR, Ellsworth PC and Ishaaya I, Biorational pest control – an 36 Byrne DN and37 Bellows MartinT,KamalA,GogoE,SaidiM,DelétréE,BonafosR TS, Whitefly biology. , , Trop www.soci.org B Dáder :acase Int J Trop Crop Prot J Agric Eng J Med Entomol Bemisia tabaci © 2014 Society of Chemical Industry :169–177 (1991). 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MI (2008). study from North America. control concept. the global dispersal of a cosmopolitan insectaphid. pest, the peach potato field crops, in Insect Sci Martin T, A temporary tunnelsmall-scale screen as farmers an to eco-friendly method protect for cabbage crops in Benin. using predators and parasitoids. Med Int Hlth acaricide-treated net to29 control phytophagous mites. impregnadas con insecticidas: unde plagas nuevo de cultivos método hortícolas. para30 el control Efficacy of mosquito netting for sustainableproduction small in holders’ Africa. cabbage screens and roof openingsRes on greenhouse ventilation. pyrethoid insecticide for treatment of mosquito nets. 39 mosquito nets. A biological test to quantify pyrethroid in impregnated nets. 3 Bethke JA and Paine TD, Screen hole size and barriers for exclusion of 1 Stern VM, Smith RF, van den Bosch R and Hagen KS, The integrated 2 Weintraub PG and Berlinger MJ, Physical control in greenhouses and 8 Martin T, Assogba-Komlan F, Sidick I, Ahle V and Chandre F, An 9 Díaz BM, Nebreda M, Salas F, Moreno A, García M and Fereres A, Mallas 4 Muñoz P, Montero JI, Antón A and Giuffrida F, Effect of insect-proof 7 Martin T, Chandre F, Chabi J, Guillet PF, Akogbeto M and Hougard JM, 5 Hougard JM, Duchon S, Zaim M and6 Guillet R, Zaim Bifenthrin: M, a useful Aitio A and Nakashima N, Safety of pyrethroid-treated 13 Blackman RL and Eastop VF, Taxonomic issues, in 14 Whalon ME, Mota-Sánchez D and Hollingworth RM, 15 Ellsworth PC and Martínez-Carrillo JL, IPM for 16 Margaritopoulos JT, Kasprowicz L, Malloch GL and Fenton B, Tracking 12 Gerling D, Alomar O and Arnó J, Biological control of 11 Licciardi S, Assogba-Komlan F, Sidick I, Chandre F, Hougard JM and 10 Martin T, Assogba-Komlan F, Houndete T, Hougard JM and Chandre F, wileyonlinelibrary.com/journal/ps REFERENCES Control of insectsystems. vectors of viruses in horticultural sustainable

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