Biological Control 41 (2007) 99–109 www.elsevier.com/locate/ybcon

Evaluation of the granulovirus and spinosad for codling moth control and impact on non-target species in orchards

Steven P. Arthurs ¤, Lawrence A. Lacey, Eugene R. Miliczky

Yakima Agricultural Research Laboratory, USDA-ARS, 5230 Konnowac Pass Road, Wapato, WA 98951, USA

Received 6 October 2006; accepted 2 January 2007 Available online 9 January 2007

Abstract

We compared the eYcacy of a commercial preparation of the codling moth, Cydia pomonella L., granulovirus, CpGV (Cyd-X®) and spinosad (Entrust®) at operational rates for codling moth control in 2004 and 2005. Concurrently we monitored the impact of treatments on populations of non-target . Spinosad was eVective at protecting fruit, with 61.6% codling moth injury in experimental plots, compared with up to 37% injury in the untreated plots at harvest. Mid-season outbreaks of the pear psylla, pyricola Foërster, were also reduced in spinosad plots. Spinosad was safe for several predators, notably the psylla predator Deraeocoris brevis Uhler, but reduced the abundance of hymenopteran parasitoids by 24% and 40% and non-target Diptera by 49% and 35%, respectively in 2004 and 2005. We found no evidence that spinosad disrupted natural control leading to increased densities of secondary pests including aphids and phytophagous mites. CpGV was less eVective than spinosad at protecting fruit, with percentage of fruits attacked similar to controls, but killed the majority (67–71%) of neonate coding moth larvae and did not harm non-target species. Additional observations were conducted in commercial orchards (mixed pear and apple) where CpGV and spinosad were used operationally against existing cod- ling moth infestations. In pear, two spray programs applied in replicated 0.4 ha blocks (i.e. CpGV followed by spinosad against the Wrst and second larval generations, respectively and vice versa) reduced fruit injury at harvest and decreased orchard pheromone monitoring trap catches by 74% over two years. In apple, CpGV was less eVective at protecting fruit in the Wrst larval generation compared with spinosad, although population suppression was eVective early in the season. Spinosad caused no disruptions of beneWcial species or sec- ondary pest outbreaks were observed in the commercial orchards. Our results suggest CpGV and spinosad can be eVectively used in inte- grated pest management for codling moth. © 2007 Elsevier Inc. All rights reserved.

Keywords: Cydia pomonella granulovirus; Toxicity; predators; Parasitoids; Integrated pest management

1. Introduction Insecticidal options for codling moth in organic and other orchards adopting low risk or integrated pest man- The codling moth, Cydia pomonella L., is the most seri- agement have historically been limited. Most ‘soft’ control ous pest of apple and a signiWcant pest of pear and walnut eVorts focused around ovicidal oils, removal of infested throughout the world (Barnes, 1991). Neonate larvae bore fruit, and more recently mating disruption (Beers et al., into the fruit and develop internally. Fifth instar larvae 1993; Calkins and Faust, 2003). The granulovirus of the leave fruit in search of cryptic habitats, such as rough bark, codling moth (CpGV) has been widely tested in North in which to spin their cocoons and pupate. In the PaciWc America (Arthurs et al., 2005; Cossentine and Jensen, 2004; Northwest there are two or three generations per year Falcon and Huber, 1991; Jaques et al., 1994; Lacey et al., (Beers et al., 1993). 2004a; Vail et al., 1991). The virus is sprayed in the orchard as an aqueous suspension to coincide with the hatching of * Corresponding author. Fax: +1 509 454 5646. eggs. Neonate larvae ingest occlusion bodies (OB), also E-mail address: [email protected] (S.P. Arthurs). called granules, before or during initial entry into fruit.

1049-9644/$ - see front matter © 2007 Elsevier Inc. All rights reserved. doi:10.1016/j.biocontrol.2007.01.001 100 S.P. Arthurs et al. / Biological Control 41 (2007) 99–109

Currently three insecticidal formulations of CpGV are reg- lure (Suterra, Bend, OR) were hung in the canopy to deter- istered for commercial use in North America (Lacey et al., mine BioWx (Wrst male catch) and monitor seasonal Xights. 2004b). Lures and sticky trap liners were changed every 2 weeks. Spinosad, comprised of a mixture of spinosyns A and D No mating disruption was used. No insecticides were used derived from fermentation of the actinomycete bacterium, apart from experimental treatments. Under-tree sprinklers Saccharopolyspora spinosa Mertz & Yao, has also recently (Nelson low angle impact model L20W) prevented insecti- been registered for orchard use in North America (Doerr cide run-oV during irrigation. Conditions were predomi- et al., 2004; Smirle et al., 2003). Two formulations of spino- nantly sunny and dry (<1 cm rainfall) throughout the study sad, one approved for organic use, are currently marketed period. In 2004 the mean temperatures was 18.4 °C (range for codling moth control. While the host range of CpGV is 3.3–35.7 °C) during the application period against the Wrst limited to codling moth and closely related species (Gröner, larval generation. During second generation tests, the mean 1990), spinosad is eVective against tortricid leafrollers and was 21.9 °C (range 8.2–37.4 °C). Temperatures in 2005 were other orchard Lepidoptera (Doerr et al., 2004; Smirle et al., similar, with means of 17.7 °C (range 2.0–35.3 °C) and 2003) and has insecticidal activity within the Diptera, Cole- 21.6 °C (range 7.4–37.0 °C) during the Wrst and second optera, Thysanoptera, and Hymenoptera (Bret et al., 1997; larval generations, respectively. Dutton et al., 2003; Thompson and Hutchins, 1999). Although CpGV might be expected to conserve natural 2.2. Experimental treatments enemies and other beneWcial organisms in the orchard agroecosystem, a disadvantage of spinosad’s broader host A commercial preparation of CpGV containing 3 £1013 range is the potential to harm beneWcial , including OB/L ‘Cyd-X®’ (Certis USA, Columbia MD) and spinosad honey and other pollinators (Mayes et al., 2003; Mor- (Entrust® Naturalyte organically approved WP formulation, andin et al., 2005), as well as some predators and parasit- Dow AgroSciences, Indianapolis IN) were applied separately oids of pests (Williams et al., 2003). Disruption in a full-season program for codling moth. Application rates of biological control may increase numbers of secondary were 220 ml/ha (Cyd-X) or 210g/ha (Entrust). The spreader- pests. In orchard tests for codling moth control, Van Stee- sticker NuFilm-17 (Miller Chemicals and Fertilizer Corp.; nwyk et al. (2005) documented elimination of the aphid Hanover, PA) was always included at 440 ml/ha. Treatments parasitoid Trixoys pallidus Haliday (no other natural ene- were timed to coincide with codling moth egg hatch using mies were reported) and increased numbers of walnut BioWx (described above) and a phenology model based on aphid Chromaphis juglandicola Kaltenbach following degree day (DD) accumulation (Beers et al., 1993). In 2004 spinosad treatment. Elevated numbers of pear rust mite initial applications were made at 282 DD post BioWx (4% egg were also reported following spinosad use in pear (Van hatch of the Wrst larval generation) with three further appli- Steenwyk and Nomoto, 2006). cations at 8 day intervals (336, 461 and 645 DD) to provide In integrated pest management (IPM) strategies, natural residual control until t95% hatch. The second larval genera- enemies (i.e. parasites, predators and pathogens) of codling tion was treated in the same way, with four applications moth and other orchard pests may play signiWcant roles in (1223, 1411, 1535 and 1738 DD). The study was repeated in crop protection (Beers et al., 1993; Cross et al., 1999; Lacey 2005, with Wve applications were made in the Wrst (243, 345, and Shapiro-Ilan, 2005). It is important to ensure that the 428, 537 and 655 DD) and four in the second (1229, 1453, various components of IPM are compatible. In this paper, 1620 and 1771 DD) larval generations. we compared the eYcacy of CpGV and spinosad at opera- The study was a randomized complete block design with tional rates for codling moth control in an experimental four (2004) or three (2005) replicate 0.04 ha (16 tree) blocks and commercial pear orchard over two years. We also mon- for each treatment including an untreated control. Repli- itored the impact of treatments on beneWcial arthropods cates were blocked in separate sections of the orchard and secondary pest outbreaks. according to tree age class. Treatments were applied in early morning during calm wind conditions (60.5 m/s) with 2. Materials and methods a diesel powered 95 L capacity airblast sprayer (HauV Company, Yakima WA) pulled with an all terrain vehicle. 2.1. Field site The sprayer was calibrated to provide full coverage of foliage and fruit at 935 L/ha at 1000 kPa. Virus was kept Orchard tests were conducted in 2004 and 2005 at the refrigerated and diluted on site. A tarpaulin screen (3 £ 9m) USDA-ARS experimental station near Moxee, WA, USA. held by four people and a buVer tree row between plots The test block was a 1 ha Bartlett pear orchard on OHXF were employed to minimize overspray or spray drift 97 rootstock at a planting density of 420 trees/ha. D’Anjou between treatments. pollinizers on the same rootstock comprised about one out of every nine trees. The orchard was originally planted in 2.3. EYcacy of CpGV and spinosad for codling moth 1987 with additional plantings in 1997, 2001 and 2002. The orchard was naturally infested with codling moth. Phero- Treatments were assessed mid season (i.e. towards the mone-baited BioLure® traps containing a 1 mg codlemone end or shortly after the Wrst larval generation; starting at S.P. Arthurs et al. / Biological Control 41 (2007) 99–109 101

783 and 1012 DD in 2004 and 2005, respectively) and simi- monitored in previous years. Each 1.6 ha section was larly before harvest after the second larval generation (1859 divided into four 0.4 ha blocks. Growers applied CpGV or and 1919 DD in 2004 and 2005, respectively). In each time spinosad in adjacent blocks; i.e. two replicate blocks each period a minimum of 150 (2004) or 300 (2005) fruit/block alternating between treatments, using a tractor-mounted were examined in situ for codling moth damage. At harvest, air-blast sprayer calibrated 1870 liter/ha at t1550 kPa. 50 infested fruit from each virus and untreated control Codling moths were monitored with pheromone traps and block were returned to the laboratory and destructively initial applications started at t250 DD (deWned previously) sampled at 10X to quantify fruit damage and larval mortal- with treatments repeated at 7–10 day intervals until t90% ity inside. The proportion of ‘deep’ larval entries (i.e. egg hatch. 76 mm) was also noted as proxy for virus concentration In pear (Hood River) there were four (2004) or Wve and speed of kill (Arthurs et al., 2005). Exit holes (made by (2005) applications of CpGV (Cyd-X at 110–220 ml/ha) or mature larvae leaving fruit) were included in the count of spinosad (Entrust WP® at 160 g/ha) in the Wrst generation, live larvae. Fruit were maintained at 12 °C for a maximum with a further three in the second generation in both years. of 2 weeks until processing. Because spinosad is restricted for resistance management (630 g/ha/season for Entrust) the test blocks were rotated in 2.4. Sampling non-target arthropods the second larval generation, i.e. treatments were spinosad followed by CpGV and vice versa. Neem oil, 1.2% Azadi- The abundance of non-target species in experimental rachtin (Aza-Direct, Gowan Co., Yuma, AZ) + 1% mineral blocks was monitored throughout the growing season. oil was also applied for psylla control in May 2005. Beating tray samples were taken approximately weekly to In apple (Mattawa) there were Wve applications of census beneWcials including predatory insects, spiders and CpGV (Carpovirusine® formulation at 1 l/ha, Arvesta parasitoids. Arboreal phytophagous species including pear Corp., San Fransisco, CA) and spinosad (Success SC® for- psylla, Cacopsylla pyricola Foërster, aphids and thrips were mulation at 438 ml/ha, Dow AgroSciences) in the Wrst gen- also noted. Arthropods dislodged by beating on two repre- eration. Eight applications of CpGV (Carpovirusine at 1 l/ sentative branches at shoulder height fell onto a canvas ha or Cyd-X at 220 ml/ha) were required in the second gen- beat tray (45 £ 45 cm). To provide a treatment buVer row eration due to protracted moth emergence from an adja- only the central four trees in each plot were sampled. Sam- cent stockpile of wooden fruit bins. Spinosad (Success is ples were taken in the cool of the morning (before 08:00 h) limited to 2.12 l/ha/season) was replaced with Wve applica- to minimize escape before identiWcation; although mobile tions of the insect growth regulator methoxyfenozide species (notably parasitoids) were still probably under-rep- (Intrepid 2F® at 1.17 l/ha + 0.5–1% oil, Dow AgroScience). resented for this reason. Parasitoids were collected with an At each location all treatment blocks (with the exception of aspirator, stored in 70% alcohol and identiWed to family. Intrepid which had fewer applications) were sprayed Foliar samples (10 randomly selected shoots/block) were concurrently or within 2 days of each other. taken bi-weekly to monitor aphid outbreaks within the Fruit evaluations for codling moth damage and beating plots. Sweep net samples were taken every 7–10 days during tray samples for non-targets were taken in the commercial spraying periods to census leafhoppers and other non-tar- orchards as previously described. Beating tray samples get taxa on the orchard Xoor. Ten sweeps along a 5 m tran- from 10–15 trees/block were taken on three occasions, early sect in the central area of each plot were used for collecting season (pre-spray), towards the end of the Wrst codling samples, which were frozen until processing. Late season moth larval generation and prior to harvest. In mixed pear populations of resident phytophagous mites were quanti- (Hood River) due to fruit russeting, pear rust mite popula- Wed from 100 randomly selected leaves in each treated and tions were also assessed (100 leaves/block from 10 trees) untreated plot. Numbers of the eriophyid pear rust mite, using a leaf brushing technique (VanBuskirk et al., 1999). Epitrimerus pyri Nalepa, were assessed using a leaf brush- Green Anjou, the most susceptible variety for both mites ing machine and the technique described by VanBuskirk and psylla, was selected for non-target assessments. No et al. (1999). mite problems were noted in apple (Mattawa).

2.5. Commercial orchard trials 2.6. Data analysis

Additional studies were conducted with cooperating Count data for experimental blocks were analyzed using growers where formulations of CpGV and spinosad were a two-factor (treatment by sample period) repeated mea- used operationally. Orchards comprised mixed pear at sures analysis of variance (ANOVA), including block irregular spaced plantings with under-tree sprinklers (Hood eVects. The abundance of non-target species was compared River, OR) and apple cv. Delicious (Mattawa, WA) at in three time periods, early, mid and late season, with sam- planting density of 426 trees/ha and overhead irrigation. In pling date the repeated factor. The analyses were done both cases experimental treatments were conWned to a 1.6 using PROC MIXED (SAS Institute, 2001). Where neces- ha section which had suVered repeated damage from cod- sary, data were normalized with log(n+1) transformation. ling moth, identiWed by the grower based on infestations SigniWcant treatment by sample period interactions in 102 S.P. Arthurs et al. / Biological Control 41 (2007) 99–109

ANOVA were further separated with simple eVects com- Table 1 ® ® parisons between pairs of means using the DIFF command EYcacy of CpGV (Cyd-X ) and spinosad (Entrust ) in Bartlett pear over and a Bonferroni adjustment for multiple comparisons. For 2 years (Moxee station) a some species, pre-treatments counts were included as covar- Treatment Mean % fruit injury § SEM iates, other species had zero or low pre-treatment counts. In 2004 2005 the commercial orchards, treatment eVects were compared Mid-season Harvest Mid-season Harvest W using one and two-way univariate ANOVA with signi cant Untreated control 0.40 § 0.5 14.8 § 4.2a 3.9 § 1.4 36.9 § 5.6a F-ratio means separated with Fisher’s LSD. In all analysis, CpGV 0.23 § 0.2 12.6 § 2.6a 1.0 § 0.5 21.8 § 6.6a signiWcance values are reported at P < 0.05. Spinosad 0.00 § 0.0 1.0 § 0.3b 1.0 § 0.9 1.6 § 0.5b Data show codling moth fruit injury in replicated 16 tree blocks. 3. Results a DiVerent letters in columns indicate diVerences (P < 0.05, 1-way contrasts in PROC GLM). 3.1. EYcacy of CpGV and spinosad on codling moth fruit in untreated control plots at harvest; 67.1 § 4% versus In both years there were distinctive peak Xights for each 20.4 § 5% (2004) and 71.3 § 2% versus 17.7 § 2% (2005), codling moth generation, (Fig. 1), which tracked the phe- respectively. The percentage of deep entries in virus-treated nology model (Beers et al., 1993). Little fruit damage was fruit was reduced compared with untreated fruit where caused by Wrst generation codling moth in either year most larvae reached the core to feed on the seeds, i.e. 51 § 6 (Table 1). Eggs were observed in plots after the Wrst Xight, versus 92 § 5 % (2004) and 52 § 4 versus 92 § 5 % (2005), but newly developing fruit was still hard and apparently respectively (P < 0.05 in independent samples t-tests). diYcult for larvae to penetrate. The second Xights were Although fruit evaluations showed the virus was more larger (Fig. 1) resulting in signiWcant fruit damage in eVective at population suppression than prevention of dam- untreated blocks at harvest (Table 1). The increased dam- age, dissections of late season fruit harvested from the virus age later in the season is illustrated by the signiWcant treat- blocks suggested possible sub-lethal eVects of the virus on W ment by generation interaction in both 2004 (F2,6 D 14.5, larval development. Fig. 2 shows a signi cantly lower P < 0.001) and 2005 (F2,4 D 15.7, P < 0.05). Spinosad was proportion of exit holes recorded from survivors in virus eVective at protecting fruit with 61.6% codling moth injury compared with untreated blocks (P D 0.011, Pearson in both years, compared with up to 37% injury in the Chi-Square test on pooled data). untreated blocks. In both years Cyd-X was signiWcantly less eVective than spinosad at protecting fruit, with percent fruit 3.2. EVects of spinosad and CpGV on non-target arthropods injury at harvest comparable with untreated controls (Table 1). However, dissections revealed most larvae in In total, 10,443 and 12,109 arthropods were collected in virus-treated fruit died near the surface in the Wrst stadium, beating trays in 2004 and 2005, respectively. Pear psylla was thus reducing the severity of injury. Larval mortality in the most abundant prey associated with populations of sev- virus-treated fruit was signiWcantly higher compared with eral beneWcial species. In 2004, most beneWcials comprised

35

30 2004 2005 25

20

15 Moths/trap

10

5

0

0 250 500 750 1000 1250 1500 1750 2000 2250 2500 Accumulated degree days

Fig. 1. Codling moth pheromone trap counts (Moxee experimental station). Data show average weekly catch from Wve (2004) or four (2005) traps against accumulated degree days (°F) post BioWx. S.P. Arthurs et al. / Biological Control 41 (2007) 99–109 103

100% occurred within spinosad-plots in 2005 (F2,7 D 9.5, P <0.05 and t 7 3.4; P < 0.05 in paired comparisons). Most of this 80% decline occurred late season, i.e. treatment by sampling W EXITS period interaction approached signi cance (F4,11 D 2.8, L5 P D 0.08). 60% L4 Other beneWcial species including ladybeetles (Cocci- L3 nellidae) and lacewings, (Chrysoperla spp., Chrysopa 40% L2 spp. and Hemerobius spp.) were recovered in all treated plots, although generally remained at low populations

Percent in category L1 20% (<1 per beat tray sample) and no signiWcant eVects of treatments were detected. Other predatory 0% including Geocoris spp. (Lygaeidae), Orius spp. (Antho- Untreated Virus plots coridae), Campylomma verbasci Meyer (Miridae) and Treatment damsel bugs (Nabidae), as well as hoverXies (Syrphidae), earwigs, ForWcula auricularia L. and the snake Xy Agulla Fig. 2. Distribution of codling moth surviving experimental treatments sp. (Neuroptera: Raphidiidae) were also recovered, among age classes (data pooled across replicates at Moxee experimental station). although these predators were too infrequent to be com- pared statistically between plots. However, when all these latter groups were pooled in the model as a single predatory insects (71.9%) followed by spiders (18.6%) and group (other predators), no signiWcant eVects of treat- parasitoids (9.5%), with similar Wndings in 2005; predatory ments were observed. insects (60.1%), spiders (20.2%) and adult parasitoids A total of 319 adult hymenopteran parasitoids among 16 (19.7%). Seasonal trends for some of the more commonly families were collected on beat trays in 2004 and 2005 recovered species are shown in Fig. 3. (Fig. 4). Overall, parasitoid abundance was reduced by In both years >95% of psylla were adults; nymphs were 23.9% and 39.2% in spinosad-plots compared with CpGV not readily dislodged. A mid-season peak (Wrst summer- and untreated plots in 2004 and 2005, respectively. This form generation), was not observed in spinosad plots in decline was statistically signiWcant in 2005, illustrated by a either year, suggesting reproductive interference (Fig. 3). treatment by sampling period interaction (F4,8 D 7.6, Repeated measures ANOVA revealed a signiWcant treat- P < 0.01) and paired comparisons revealed the reduction in ment by sample period interaction for psylla in 2004 parasitoids in spinosad plots occurred mid-season (t 7 3.2; (F4,16 D 5.5, P < 0.01) and paired comparisons revealed that P < 0.05 in paired comparisons). We did not quantify the psylla abundance was reduced during the mid-season in impact on diVerent parasitoid guilds, although 57% of col- spinosad plots compared with untreated or CpGV-treated lected individuals were the encyrtid Trechnites insidiosus plots (t 7 3.3; P < 0.05). Although the treatment by sample Crawford, an important parasitoid of psylla (Beers et al., period interaction was not statistically signiWcant in 2005 1993). (F4,18 D 2.0, P D 0.18), a similar trend was observed. No secondary pest outbreaks occurred in our plots. A Deraeocoris brevis Uhler (Heteroptera: Miridae), was the mid-season infestation of Aphis pomi De Geer was noted in most abundant beneWcial species and comprised 79.4% and one of the spinosad-plots, but was quickly brought under 74.7% of all insect predators in 2004 and 2005, respectively. control by beneWcials. Most aphids were alates (overall 74% The abundance of D. brevis increased mid-season and was in 2004) indicating limited reproduction. Other foliar pests not reduced by spinosad treatments in either year. A signiW- including western tentiform leaf miner, Phyllonorycter cant treatment by sampling period interaction in 2004 elmaella Doganlar & Mutuura, stink bugs Euschistus con- (F4,12 D 3.7, P < 0.05) was caused by more D. brevis in spino- spersus Uhler and Acrosternum hilare Say (Hemiptera: sad-plots late season compared with other plots (t 7 3.3; Pentatomidae), lygus bug, mainly Lygus lineolaris Palisot P < 0.05). There was thus circumstantial evidence D. brevis de Beauvois and the white apple leafhopper Typhlocyba was largely responsible for the late season decline in psylla pomaria McAtee were found, but remained below damag- populations. Another psylla predator, Anthocoris spp., ing thresholds (<1 per beat tray sample) in all plots, and no mainly Anthocoris tomentosus Péricart (Heteroptera: statistically signiWcant treatment eVects were observed for Anthocoridae), also increased mid-season. Unlike D. brevis, these species. Thrips were a minor pest (63 per beat tray) the increased numbers of Anthocoris spp. were not observed post-bloom when treatments were applied, although Frank- in spinosad plots in either year, although the relatively liniella occidentalis Pergande were collected throughout the small sample size prevented treatment eVects from being season. Thrips were maintained at low levels (<1 per beat statistically signiWcant. A large diversity of arboreal spiders tray) with spinosad treatments and signiWcant treatments V was recovered from the foliage in both years; common fam- e ects were observed in 2004 (F2,7 D 8.1, P < 0.05) caused by ilies included Linyphiidae, Salticidae, Oxyopidae and fewer thrips within spinosad compared with CpGV or Clubionidae, although species were not identiWed. No treat- untreated plots mid season (t 7 3.0; P < 0.05 in paired ment eVects were observed in 2004 although fewer spiders comparisons). 104 S.P. Arthurs et al. / Biological Control 41 (2007) 99–109

35 70 Pear psylla, 2004 2005 30 60 Untreated CpGV 25 50 Spinosad 20 40 15 30 10 20 5 10 0 0

14 14 D. brevis, 2004 2005 12 12 Untreated CpGV 10 10 Spinosad 8 8 6 6 4 4 2 2 0 0

2.0 1.0 Anthocoris spp. 2004 2005 Untreated 0.8 1.5 CpGV Spinosad 0.6 1.0 0.4 0.5 0.2

0.0 0.0

4 4 Spiders, 2004 2005 Untreated 3 3 CpGV

Number of individuals per beating tray sample Spinosad

2 2

1 1

0 0

3.5 6 Parasitoids, 2004 2005 3.0 5 Untreated CpGV 2.5 4 Spinosad 2.0 3 1.5 2 1.0 0.5 1 0.0 0 145 152 161 168 173 183 189 196 207 214 222 229 238 246

118 125 132 139 146 153 160 167 174 188 202 209 216 223 230 237 244 252 262 269 Julian day Julian day

Fig. 3. Pear psylla, Cacopsylla pyricola, and common beneWcial taxa monitored with beating trays over two years (Bartlett pear, Moxee experiment station). Data show means § SEM for four (2004) or three (2005) replicate 16 tree blocks. Arrows show timing of spray treatments. S.P. Arthurs et al. / Biological Control 41 (2007) 99–109 105

200

2004 2005 150

100 Number individuals 50

0 Figitidae Eucoilidae Torymidae Encyrtidae Charipidae Mymaridae Eulophidae Braconidae Scelionidae Aphelinidae Eurytomidae Megaspilidae Pteromalidae Platygastridae Ceraphronidae Ichneumonidae

Fig. 4. Distribution of parasitoids collected in beat trays from Bartlett pear over two years (Moxee experiment station, pooled among diVerent experimen- tal plots).

Overall 4660 and 26,007 arthropods were collected in 35.3 § 8.7% (2005) in spinosad-treated compared with other sweep net samples in 2004 and 2005, respectively (in 2004 plots (Fig. 5). Repeated measures analysis using pre-treat- sampling was more limited around spraying periods). Leaf- ment covariates and two (2004) or three (2005) sampling hoppers (Homoptera: Cicadellidae) comprised the majority periods showed the decline in non-target dipterans in spino- W W of epigeal taxa, 49% in 2004 and 88% in 2005. No signi - sad plots was signi cant in 2004 (F2,8 D 15.6, P < 0.01 and cant treatment eVects were observed for leafhopper abun- t 7 4.4, P < 0.01 in paired comparisons) and 2005 dance in 2004 (all species pooled) or 2005 (major species (F2,3 D 13.7, P < 0.05 and t 7 4.4; P < 0.05). Species mainly tested separately were Dikraneura spp. and Ceratagalia spp. comprised fungus gnats (Mycetophilidae) and aphid Xies or all unidentiWed nymphs pooled). However the abun- (Chamaemyiidae) but were not further identiWed. dance of non-target dipterans (representing 47% and 11% Late season leaf assessments in both years revealed high of arthropods collected in sweep nets 2004 and 2005, populations of pear rust mites, although mite abundance respectively) was reduced by 48.8 § 3.8% (2004) and was not increased by spinosad treatments (Table 2). The

80 80 , 2004 2005 Untreated 60 60 CpGV Spinosad

40 40

20 20

0 0 Number flies per sweep net sample 140 150 160 170 180 190 200 210 220 230 240 250 260 132 139 146 153 160 167 174 188 202 209 216 223 230 237 244 252 269 Julian day Julian day

Fig. 5. Non-target Diptera (species pooled, see text) collected in sweep net samples over two years (Bartlett pear, Moxee experiment station). Data show means § SEM for four (2004) or three (2005) replicate 16 tree blocks. Arrows show timing of spray treatments. 106 S.P. Arthurs et al. / Biological Control 41 (2007) 99–109

Table 2 orchard there was a relatively high initial codling moth Tests for late season mite resurgence in Barlett pear (Moxee station) infestation, with Bartlett the more susceptible variety com- Treatment Number PRM/leaf § SEM (n D 100 leaves/block) pared with Anjou (Table 3). Both spray programs (i.e. 2004 2005 CpGV followed by spinosad against the Wrst and second V Upper canopy Lower canopy Terminal leaf Spur leaf generations, respectively, and vice versa) were e ective at reducing fruit injury at harvest over two years. Lowest Untreated 103.8 § 40.1 38.6 § 12.6 203.8 § 31.3 78.2 § 16.7 CpGV 101.7 § 46.0 85.0 § 19.3 305.5 § 21.9 75.2 § 19.8 damage occurred when spinosad was applied against the Spinosad 90.4 § 8.5 70.2 § 23.3 230.8 § 32.8 82.3 § 3.5 Wrst larval generation. Seasonal pheromone trap catches in Pear rust mite (PRM), Epitrimerus pyri, was evaluated over two years with the orchard (average of four traps maintained in the same diVerent methods. locations) were reduced from a pre-treatment catch of 111.3 Means in column not signiWcantly diVerent (P > 0.05, 1-way ANOVA). in 2003 to 68.8 in 2004 (38% reduction) and 29.3 in 2004 (74% reduction). Larval mortality varied from 63 to 90% in Table 3 Y ® ® virus-sprayed fruit in the individual blocks. The grower E cacy of CpGV (Cyd-X at 110–220 ml/ha) and spinosad (Entrust at V 140–210 g/ha) for codling moth control in mixed pear (Hood River) considered the treatments e ective and reduced virus appli- b cation rates to 110 ml/ha during 2005. In apple, spinosad Spray program Percent fruit injury § SEM V W 1st/2nd generationa was more e ective at protecting fruit in the rst larval gen- cv. 2004 2005 eration compared with CpGV (Table 4). Nevertheless, Mid-season Harvest Mid-season Harvest examination of infested fruit revealed >92% of larvae in the CpGV/spinosad Bartlett 8.3 § 2.2a n/a 1.4 § 0.1 1.2 § 0.5 CpGV plots were killed, indicating eVective population Anjou 1.9 § 0.5b 1.1 § 0.1 0.5 § 0.1 n/a suppression early in the season. CpGV was less eVective Spinosad/CpGV Bartlett 6.2 § 2.0a n/a 0.7 § 0.1 0.5 § 0.2 again the second larval generation and killed signiWcantly Anjou 0.9 § 0.1b 2.4 § 1.4 0.0 § 0.0 n/a fewer larvae compared with spinosad/methoxyfenozide, The order in which treatments were applied was rotated between the Wrst although fruit injury at harvest was similar between blocks and second larval generations. treated with either program (Table 4). n/a, not assessed. a Applied at 1870 l/ha in two replicate blocks each 0.4 ha. All plots In total, 2383 and 1276 arthropods were collected in treated with neem oil (Azadirect) + 1% mineral oil in May 2005. beating trays from Green Anjou (Hood River) in 2004 and b A minimum of 300 fruit/block evaluated; diVerent letters indicate 2005, respectively, with a further 390 from apple in 2004 signiWcant diVerences (P < 0.05, Fisher’s LSD). (Mattawa). Pear psylla was the most common species in pear, with average mid-season counts of 14.5 (2004) and Table 4 EYcacy of CpGV (Carpovirusine® at 1 l/ha or Cyd-X® at 220 ml/ha) 12.3 (2005) per beat tray sample. D. brevis was the most versus spinosad (Success SC® at 438 ml/ha) all applied at 1870 l/ha in common predator, averaging 3.4 (2004) and 1.6 (2005) indi- apple in 2004 (Mattawa) viduals per beat tray later in the season. In both years psylla Spray program Fruit injury and larval mortality %§SEMb infestations in all blocks declined to low levels (<2 per beat 1st/2nd generationa Mid-season Harvest tray) by late season, ostensibly largely due to predation by D. brevis. Other predators included Anthocoris spp., spiders, Injury Mortality Injury Mortality coccinellids, lacewings, Orius spp., earwigs and also parasit- CpGV 3.3 § 1.6 92.5 § 7.4 3.5 § 1.5 65.9 § 2.8a oids; although apart from spiders in 2004 these groups § § § § Spinosad/methoxyfenozide 0.8 0.3 80.0 20.0 4.1 1.1 81.9 0.3b comprised <1 per beat tray sample. A fairly large F. occi- Data show fruit injury and larval mortality in sprayed fruit. Spinosad was ® dentalis population (24/beat tray) was also observed in replaced with methoxyfenozide (Intrepid 2F at 1.2 l/ha) in the second apple early season (pre-spray). No other secondary pest generation. a Applied at 1870 l/ha in two replicate blocks each 0.4 ha. problems were observed in any blocks in either orchard and b A minimum of 450 (mid-season) or 1080 (harvest) fruit/block no signiWcant diVerences (P > 0.05) were observed in species evaluated; diVerent letters indicate signiWcant diVerences (P < 0.05, inde- abundance between the diVerent plots for any sample dates. pendent T-test). Late season leaf assessments in 2004 revealed high popula- tions of E. pyri, although mite abundance was similar predatory mite Typhlodromus spp. and the mite feeding between diVerent spray programs (Table 5). beetle Stethorus picipes Casey (comprising 85% and 47% of Table 5 all coccinellids observed in 2004 and 2005, respectively) Tests for late season mite resurgence in 2004 were observed in all experimental plots. Spray program 1st/2nd Number PRM/10 leaves § SEMb generationa 3.3. Commercial orchard trials Terminal leaf Spur leaf CpGV/spinosad 90.1 § 22.6 22.0 § 10.9 Fruit evaluations for codling moth control are shown in Spinosad/CpGV 118.0 § 17.0 26.7 § 10.4 Tables 3 and 4. Both orchards experienced a signiWcant Pear rust mite (PRM), Epitrimerus pyri, was evaluated on Green Anjou infestation, coming from adjacent orchard habitat (Hood (Hood River). a Applied in 0.4 ha blocks replicated twice. River) while a large fruit bin pile (Mattawa) also provided a b 100 leaves/block sampled; means in column not signiWcantly diVerent constant source of immigrating moths. In the mixed pear (P > 0.05, 1-way ANOVA. S.P. Arthurs et al. / Biological Control 41 (2007) 99–109 107

4. Discussion D. brevis, a key pear psylla predator in the region, appeared compatible with spinosad in both experimental (Fig. 3) and 4.1. EYcacy of CpGV and spinosad commercial pear orchards (Hood River). At Moxee, there was evidence that spinosad reduced spiders, Anthocoris spp. We noted that CpGV and spinosad controlled codling and parasitoids, although the relative abundance of these moth, both in experimental blocks (Table 1) and in com- groups may have been confounded by the reduced prey mercial orchards faced with economically damaging infes- (psylla) in spinosad-treated blocks. On the other hand, the tations (Tables 3 and 4). Moth catch data from Hood River small plots might have allowed immigration and the delete- suggested two or more years of treatments may be required rious impact of spinosad treatments (especially for more to bring populations under control. However, increased mobile parasitoids) could be more pronounced in a large fruit injury was observed in CpGV-treated plots compared orchard. Spinosad also had activity against psylla, thrips with spinosad (Table 1). This diVerence is explained by and non-target dipterans (Figs. 3 and 5). diVerent modes of action. CpGV has no contact activity Our Weld data support a series of laboratory bioassays and must be ingested. The virions (virus DNA) pass that documented no acute toxicity of a Weld rate of spino- through the mid-gut peritrophic membrane, invade and sad for several orchard predators, Chrysoperla carnea, multiply in body tissues including the tracheal matrix and F., Galendromus (D Typhlodromus) fat body (Federici, 1997). Death occurs in 5–10 days, allow- occidentalis Nesbitt (western predatory mite) and D. brevis ing infected larvae to cause shallow entries or ‘stings’ in (Unruh et al., 2006). However, the Weld rate of spinosad fruit. Most orchardists have a low tolerance for damage (and several other nominally more selective new insecti- (typically 1%), although fruit with superWcial damage may cides) caused reduced fecundity in all species. Other be suitable for processing for juice or canning. However, sub-lethal eVects included reduced egg hatch (C. carnea and CpGV may also cause delayed development or sub-lethal D. brevis) and reduced female longevity and oVspring sur- eVects (Fig. 2), suggesting CpGV may have a subtle and vival (A. nemoralis) (Unruh et al., 2006). Two species of longer term impact on pest populations. Biache et al. (1998) parasitoids, including the codling moth larval parasitoid reported a 25% reduction in adult emergence among cod- Mastrus ridibundus Gravenhorst, and the European earwig ling moth larvae surviving treatment with Carpovirusine. F. auricularia L. also suVered high mortality at the Weld Spinosad is a neurotoxin that demonstrates rapid contact rate, while both parasitoids also suVered sub-lethal eVects and ingestion activity in insects, causing excitation of the at a reduced (10%) rate (Unruh et al., 2006). nervous system, leading to cessation of feeding and paraly- A literature survey by Williams et al. (2003) concluded sis (Salgado, 1998). Symptoms of poisoning are consistent that hymenopteran parasitoids are signiWcantly more sus- with activation of nicotinic acetylcholine receptors, but ceptible to spinosad than predatory insects. Using the Inter- caused by a mechanism that is novel among known insecti- national Organization for Biological and Integrated cide compounds; spinosad acts at a diVerent site than imi- Control of Noxious and Plants (IOBC) criteria for dacloprid and other nicotinic receptor-based insecticides both laboratory and Weld tests that run from one (harmless) (Salgado, 1998). Table 1 indicates spinosad killed or to four (harmful) (Hassan, 1992; Sterk et al., 1999), overall, incapacitated larvae before they damaged fruit. 71% (42/59) of laboratory studies and 79% (81/103) of Weld- Although we made no direct comparisons between crops type studies on predators (27 species) gave a class 1 result in these studies, it was noted that codling moth mortality with spinosad. Coccinellid species, the anthocorid Orius rates in our virus-treated experimental pear orchard (e.g. insidiosus Say, the hemipteran Geocoris punctipes Say and 67–71% in 2004 and 2005) were lower compared with previ- the chrysopids Chrysoperla ruWlabris Burmeister and ous studies in apple where equivalent virus treatments rou- C. carnea Stephens were considered particularly tolerant tinely killed 790% larvae, with fewer deep entries in (Williams et al., 2003). The earwig, Doru taeniatum Dohrn infested fruit (Arthurs et al., 2005; Lacey et al., 2004a). The (Cisneros et al., 2002) and some syrphid Xies (Michaud, eVectiveness of CpGV on apple and pear may reXect diVer- 2003; Torres et al., 1999; Vinuela et al., 2001) were suscepti- ences in deposition, persistence, or activity of virus at the ble to Weld rates of spinosad in laboratory tests. By contrast leaf or fruit surface. spinosad showed direct toxicity and sublethal eVects (e.g. loss of reproductive capacity) to a wide range of parasitoids 4.2. Non-target eVects of spinosad and CpGV (25 species), with 78% (35/45) of laboratory studies and 86% (18/21) of Weld-type studies returning a moderately Spinosad has been classiWed as an environmentally and harmful or harmful result (Williams et al., 2003). toxicologically reduced risk pesticide by the United States Despite any harmful eVect on parasitoids, we found no Environmental Protection Agency. It degrades quickly and evidence that spinosad disrupted natural control leading to generally shows little residual insecticidal activity 3–7 days outbreaks of secondary pests. High populations of phy- after application (Williams et al., 2003). However, docu- tophagous mites, predominantly E. pyri, were observed mentation of the spinosad’s harmful eVects on several both in small experimental (Moxee) and larger orchard beneWcial species has raised concern over its compatibility blocks (Hood River), but not exacerbated by spinosad in with biological pest management. In the present studies either case (Tables 2 and 5). Regionally, predatory mites, 108 S.P. Arthurs et al. / Biological Control 41 (2007) 99–109 chieXy Typhlodromus spp., provide eVective control of Acknowledgments E. pyri (Beers et al., 1993) and Typhlodromus caudiglans Schuster was noted from spinosad plots during mite count- We thank Heather Headrick, Dan Hallauer, Kristina ing. Previous Weld studies reported operational rates of Campbell, Amy Vangstad, Chey Temple, Dana Jones, spinosad had no eVect on predatory phytoseiid mites, PerriAnn Halbert, Jonathan Pruneda, Lisa Hannigan and including T. pyri (Miles and Dutton, 2003), although spino- Belinda Bray-Bishop for assistance with these studies. We sad was reported harmful to adult Neoseiulus fallacis Gar- also thank Jerry Gefre and John Harvey (Moxee experi- man (Villanueva and Walgenbach, 2005). However, caution ment station) and our grower cooperators Bill Mellow, in using spinosad is prudent in commercial orchards where Orlin Knutson and Naná Simone for technical support. few or no alternative predators are present. As noted ear- Rob Fritts Jr. (Certis USA) and Harvey Yoshida (Dow lier, Van Steenwyk et al. (2005) and Van Steenwyk and AgroScience) kindly provided materials used in the studies. Nomoto (2006) documented increased numbers of aphids We are very grateful to Bruce Mackey (Statistician, PaciWc and mites in orchard tests following spinosad use. West Area, USDA-ARS) and David Horton (USDA-ARS) for assistance with data analysis and Don Hostetter 5. Conclusions (Hostetter consulting Intl. Granville, OH) and Wee Yee (USDA-ARS) for constructive reviews of the manuscript. CpGV and spinosad oVer new options to manage cod- The Washington Tree Fruit Research Commission ling moth in organic or ‘soft’ orchards, although their short provided Wnancial support. residual activity may limit adoption in conventional insecti- cide programs. The limited protection to fruit also makes References CpGV less eVective under high pest pressure. Our data sug- gest both products are compatible with integrated pest Arthurs, S.P., Lacey, L.A., Fritts, R., 2005. 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