Lethal and Sublethal Impacts of Acaricides on Tamarixia Radiata (Hemiptera: Eulophidae), an Important Ectoparasitoid of Diaphorina Citri (Hemiptera: Liviidae)

Journal of Economic Entomology Advance Access published July 3, 2015

ECOTOXICOLOGY Lethal and Sublethal Impacts of Acaricides on Tamarixia radiata (Hemiptera: Eulophidae), an Important Ectoparasitoid of Diaphorina citri (Hemiptera: Liviidae)

A. C. S. LIRA,1 O. Z. ZANARDI,2 V. H. BELOTI,2 G. P. BORDINI,2 P. T. YAMAMOTO,2,3 J. R. P. PARRA,2 1 AND G. A. CARVALHO

J. Econ. Entomol. 1–11 (2015); DOI: 10.1093/jee/tov189 ABSTRACT The use of synthetic acaricides for management of pest mites may alter the efficacy of the ectoparasitoid Tamarixia radiata (Waterston) in biological control of Diaphorina citri Kuwayama, the vector of the bacteria associated with huanglongbing (HLB) in citrus orchards. We evaluated the toxicity of 16 acaricides that are recommended for the control of citrus-pest mites to T. radiata. Acrinathrin, bifenthrin, carbosulfan, and fenpropathrin caused high acute toxicity and were considered harmful (mortality >77%) to T. radiata. Abamectin, diflubenzuron, etoxazole, fenbutatin oxide, fenpyroximate, flufenoxuron, hexythiazox, propargite, spirodiclofen, and sulfur caused low acute toxicity and affected the parasitism rate and emergence rate of adults (F1 generation), and were considered slightly harmful to T. radiata. Dicofol and pyridaben did not affect the survival and action of the ectoparasitoid, and were considered harmless. In addition to its acute toxicity, carbosulfan caused mortality higher than 25% for >30 d after application, and was considered persistent. Acrinathrin, bifenthrin, fenpropathrin, fenpyroximate, propargite, and sulfur caused mortalities over 25% until 24 d after application and were considered moderately persistent; abamectin was slightly persistent, and fenbutatin oxide was short lived. Our results suggest that most acaricides used to control pest mites in citrus affect the density and efficacy of T. radiata in the biological control of D. citri. However, further evaluations are needed in order to deter- mine the effect of these products on this ectoparasitoid under field conditions.

KEY WORDS biological control, acaricide, mortality, parasitism rate, emergence rate

Huanglongbing (HLB) is among the most important Therefore, conservation and augmentation of biologi- and destructive citrus diseases worldwide (Bove´2006, cal control agents are important strategies to reduce Grafton-Cardwell et al. 2013). In Brazil, HLB is pre- population levels of pests and the impacts caused by sent in 6.9% of cultivated citrus trees (200 million overuse of insecticides in citrus orchards. Among the trees) in the state of Saõ Paulo, the main citrus-produc- natural enemies, the ectoparasitoid wasp Tamarixia ing region of the country (Fundecitrus 2013). Given radiata (Waterston) (Hemiptera: Eulophidae) has the absence of a cure, HLB is managed by planting shown great potential for use in pest management pro- healthy seedlings, reducing inoculum by eliminating grams to control D. citri (Pluke et al. 2008, Qureshi symptomatic trees, and mainly, control of the insect et al. 2009, Hall and Nguyen 2010, Williams et al. vector, the Asian citrus psyllid Diaphorina citri 2013). T. radiata is a specialized ectoparasitoid, devel- Kuwayama (Hemiptera: Liviidae). Currently, in Brazil, oping preferentially in third- to fifth-instar D. citri growers are spraying insecticides (organophosphates, nymphs (Skelley and Hoy 2004, Hall et al. 2013). In pyrethroids, and neonicotinoids) from 18 to 25 times Brazil, this ectoparasitoid was found in 2006 in the mu- annually, to control D. citri (Belasque Jr. et al. 2010). nicipality of Piracicaba, Saõ Paulo, and is currently pre- Despite their high efficacy, these products may alter sent in most citrus-producing regions of the country the balance between pests and their natural enemies, (Paiva and Parra 2012). It can be mass-reared in the causing outbreaks of secondary pests, resurgence of tar- laboratory for inundative release in field conditions, get pests, and selection of resistant populations (Yama- and rapidly suppresses populations of D. citri (Chen moto and Bassanezi 2003, Tiwari et al. 2011, Guedes and Stansly 2014). In recent years, successive releases and Cutler 2013). of T. radiata have been conducted in areas of HLB management programs, as well as in areas with orange jasmine [Murraya paniculata (L.) Jack (Rutaceae)] infested with D. citri nymphs or in areas next to commer- 1 Department of Entomology, Federal University of Lavras cial orchards, where the ectoparasitoid is multiplying (UFLA), 37200-000, Lavras, MG, Brazil. (Parra et al. 2010). In Saõ Paulo state, early estimates 2 Department of Entomology and Acarology, College of Agriculture “Luiz de Queiroz”/University of Saõ Paulo (ESALQ/USP), 13418-900, of parasitism rates ranged from 27.5 to 80% (Go´mez- Piracicaba, SP, Brazil. Torres et al. 2006, Parra et al. 2010). However, with in- 3 Corresponding author, e-mail: [email protected]. creased use of chemical pesticides for control of the

VC The Authors 2015. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: [email protected] 2JOURNAL OF ECONOMIC ENTOMOLOGY vector D. citri, parasitism rates were reduced to below Pesticides. Sixteen commercial acaricides registered 25.7% (Paiva and Parra 2012). for the control of pest mites (especially B. phoenicis In addition to D. citri, the flat mite Brevipalpus and P. oleivora) in citrus orchards were evaluated on phoenicis (Geijskes) (Acari: Tenuipalpidae), vector of T. radiata. All compounds were tested without adjuvant the Citrus Leprosis Virus (CiLV), and the citrus rust at the label rates recommended by the Brazilian Minis- mite Phyllocoptruta oleivora (Ashmead) (Acari: Eryo- try of Agriculture, Livestock and Food Supply (MAPA; phyidae) are considered key pests of this crop (Oliveira Agrofit 2014). The acaricides and concentrations (g a.i. and Pattaro 2008). Although less important, mites of liter1) used in the bioassays are described in Table 1. the family Tetranychidae, especially the citrus red mite Bioassays. All laboratory bioassays were conducted Panonychus citri (McGregor) and the Texas citrus mite in a climate-controlled room (25 6 2C, 70 6 10% RH, Eutetranychus banksi (McGregor), also require control and a photoperiod of 14:10 [L:D] h), using a fully because of the continual population outbreaks observed randomized design. after successive applications of insecticides to control Bioassay 1: Acute Toxicity of Acaricides to D. citri (Yamamoto and Zanardi 2013). This increased T. radiata Adults. To assess the acute toxicity of acari- use of acaricides can alter biological and behavioral pa- cides to T. radiata adults, leaf discs (3.3 cm in diameter) rameters of the ectoparasitoid T. radiata and affect the of Valencia sweet orange [Citrus sinensis (L.) Osbeck action of this natural enemy in controlling D. citri. (Rutaceae)] were initially sprayed with 2 ml of solution, Some studies have demonstrated the acute and residual in a Potter tower (Burkard Scientific, Uxbridge, United toxicity of certain acaricides to T. radiata (Cocco and Kingdom) adjusted to a pressure of 0.7 kg cm2, result- Hoy 2008, Hall and Nguyen 2010, Lira et al. 2014), but ing in a fresh dry deposition of 1.8 6 0.1 mg cm2, none has examined the sublethal effects of acaricides according to the criteria established by the Pesticides on its biological parameters and parasitism efficiency. and Beneficial Organisms Working Group of the Inter- Knowledge of sublethal effects contributes to the un- national Organization for Biological Control of Noxious derstanding of the impacts of pesticides on natural ene- Animals and Plants, West Palearctic Regional Section mies (Desneux et al. 2007). This information is (IOBC/WPRS) for studies of pesticide toxicity to natu- important for the development of management strate- ral enemies (Van de Veire et al. 2002). Distilled water gies aimed at the conservation and augmentation of bi- was used as a control treatment. ological control agents, and also to guarantee the After the treatments were sprayed, the discs were success of integrated pest management (IPM) pro- kept in a climate-controlled room for 3 h to allow the grams in citrus. Considering the importance of T. radi- residues to dry. Then, the discs were placed in individ- ata as a biological control agent of D. citri,thisstudy ual Petri dishes (3.5 cm in diameter by 0.7 cm in assessed the lethal and sublethal impacts of 16 acari- height) containing a 2-mm layer of agar: water (2.5% cides that are recommended for the control of citrus w/v)andusedasanexperimentalunit.Next,10parasi- mites, on this ectoparasitoid. toid adults (5 females and 5 males) up to 24 h old were anesthetized with CO2 for 5 s and placed in each experimental unit. The experimental units were sealed with Materials and Methods voile fabric to allow gas exchange and prevent the accu- Rearing of T. radiate. The culture was established mulation of excess moisture. A honey droplet (1 from adults collected in citrus orchards in the munici- mm3) was placed on the voile to serve as food for the pality of Piracicaba, Saõ Paulo, Brazil. The insect popu- parasitoids during the assay period. For each treatment, lations were maintained at the Insect Biology five repetitions were used. Laboratory of the Department of Entomology and The number of alive and dead insects was recorded Acarology of the “Luiz de Queiroz” College of Agricul- 24 h after exposure to the residues. Moribund parasi- ture, University of Saõ Paulo (ESALQ/USP), Piraci- toids and those that did not react to the touch of a fine caba, Saõ Paulo, Brazil. The culture was maintained in brush were considered dead. The acute toxicity (mor- a climate-controlled room (25 6 2C, 70 6 10% relative tality) of each acaricide was calculated using the for- humidity [RH], and a photoperiod of 14:10 [L:D] h). mula of Abbott (1925). Based on mortality data (M), For the rearing we used seedlings of orange jasmine the acaricides were classified according to the criteria (M. paniculata) cultivated in pots (2 liter). The seed- defined by the IOBC/WPRS (Van de Veire et al. 2002): lings were initially pruned to a height of 25 cm, and class 1 ¼ harmless (M < 25%), class 2 ¼ slightly harmful after producing sprouts (2–3 cm in length), they were (25% M 50%), class 3 ¼ moderately harmful placed in rearing cages (40 by 60 by 50 cm3) and used (51% M 75%), and class 4 ¼ harmful (M > 75%). as a substrate for oviposition of females and feeding by Bioassay 2: Sublethal Effects of the Acaricides nymphs and adults of D. citri, according to the method on T. radiata Adults. The acaricides considered proposed by Nava et al. (2007).WhenD. citri nymphs harmless (class 1) in the evaluation of acute toxicity reached the fourth and fifth instars, the seedlings were (bioassay 1) were used in the second bioassay. For this, transferred to acrylic cages (90 by 50 by 50 cm3)and 20 females of T. radiata up to 24 h old were released in T. radiata adults were released to parasitize the each experimental unit made with Petri dishes (3.5 cm nymphs, according to the method described by in diameter by 0.7 cm in height) containing a leaf disc Go´mez-Torres et al. (2012). Drops of honey were (3.3 cm in diameter) of Valencia sweet orange treated placed on the sides of the cages as food for the with the respective acaricide or deionized water parasitoid adults. (control) and placed on a 2-mm layer of agar: water 2015

Table 1. Acaricides used in the bioassays L R TAL ET IRA

Common name Trade name (MoA IRAC)a Chemical group Dosage used Manufacturer Target pestb Other pests controlledc (g a. i. liter1) :E .: Abamectin Vertimec 1.8 EC (6) Activators of chloride channels 0.005 Syngenta CRM1 CRM2, TCM, BM, CL, ACP Acrinathrin Rufast 5 SC (3A) Sodium channel modulators 0.005 Bayer FM – FC OF FFECT Bifenthrin Talstar 10 EC (3A) Sodium channel modulators 0.020 FMC FM CRM1, CFB, CO, ACP Carbosulfan Marshal Star 70 EC (1A) Acetylcholinesterase inhibitors 0.350 FMC CRM1 ACP Dicofol Dicofol 18.5 EC (UN) Compounds of unknown or uncertain MoA 0.370 Milenia FM CRM1 Diflubenzuron Micromite 24 SC (15) Inhibitors of chitin biosynthesis, type 0 0.120 Chemtura CRM1 CL, CFB, ACP Etoxazole Borneo 4.6 SC (10B) Mite growth inhibitors 0.021 Sumitomo FM – A 16 Fenbutatin oxide Torque 50 SC (12B) Inhibitors of mitochondrial ATP synthase 0.400 Basf FM BM, CRM1, CRM2, TCM Fenpropathrin Danimen 30 EC (3A) Sodium channel modulators 0.150 Sumitomo FM BCA, CFB, MFF, CO, ACP ON CARICIDES Fenpyroximate Ortus 5 SC (21A) Mitochondrial complex I electron transport inhibitors 0.050 Arysta CRM1 BM, FM Flufenoxuron Cascade 10 EC (15) Inhibitors of chitin biosynthesis, type 0 0.050 Basf CRM1 BM, FM Hexythiazox Savey 50 WP (10A) Mite growth inhibitors 0.015 DuPont FM – Propargite Omite 72 EC (12C) Inhibitors of mitochondrial ATP synthase 0.720 Chemtura CRM1 CRM2, FM, TCM Pyridaben Sanmite 20 EC (21A) Mitochondrial complex I electron transport inhibitors 0.150 Iharabras FM CRM1

Spirodiclofen Envidor 24 SC (23) Inhibitors of acetyl CoA carboxylase 0.060 Bayer CRM1 BM, CRM1, FM, TCM radiata Tamarixia Sulfur Kumulus DF 80 WG MAD 4.000 Basf FM BM, CRM1 a Classification of the mode of action of pesticides according to Insecticide Resistance Action Committee (IRAC, 2014); MAD, mechanism of action not defined. b Target pests: CRM1, citrus rust mite [Phyllocoptruta oleivora (Ashmead)]; FM, flat mite [Brevipalpus phoenicis (Geijskes)]. c Other pests controlled: CRM2, citrus red mite [Panonychus citri (McGregor)]; TCM, Texas citrus mite [Eutetranychus banksi (McGregor)]; BM, broad mite [Polyphagotarsonemus latus (Banks)]; CL, citrus leafminer (Phyllocnistis citrella Station); CFB, citrus fruit borer (Gymnandrosoma aurantianum Lima); CO, citrus orthezia [Praelongorthezia praelonga (Douglas)]; MFF, Mediterranean fruit fly [Ceratitis capitata (Wiedemann)]; BCA, brown citrus aphid [Toxoptera citricida (Kirkaldy)]; and ACP, Asian citrus psyllid (Diaphorina citri Kuwayama). 3 4JOURNAL OF ECONOMIC ENTOMOLOGY

(2.5% w/v). Females were kept on product residues for integrated pest management programs. For this, Rang- a period of 24 h. For each treatment, five repetitions pur lime seedlings cultivated in 100-ml pots were were used (n ¼ 100). After this period, the moribund pruned to a height of 25 cm to stimulate growth. After and dead females were counted (acute toxicity) and the sprouts (2–3 cm in length), each seedling was infested surviving females were transferred to glass cages (90 by with 20 D. citri nymphs of fourth and fifth instars and 50 by 50 cm3) containing Rangpur lime seedlings placed in a climate-controlled room for 24 h to allow (15 cm in height) infested with D. citri nymphs in the the nymphs to acclimate on the host plant. After this fourth and fifth instars, in a proportion of a female of period, the seedlings were placed in glass cages (90 by the ectoparasitoid to 10 D. citri nymphs (daily parasit- 50 by 50 cm3)andT. radiata females were added to ism rate of 7 D. citri nymphs per T. radiata female allow them to parasitize the nymphs (proportion of one [Go´mez-Torres 2009]). The parasitism period was 48 h. female of ectoparasitoid to 10 D. citri nymphs). The After this period, the seedlings were transferred to new exposure time for the parasitism was 24 h. Then, the cages (90 by 50 by 50 cm3) to evaluate the parasitism seedlings with the parasitized nymphs were transferred rate. The number of nymphs parasitized in each treat- to new glass cages (90 by 50 by 50 cm3), where they ment was assessed 9 d after the seedlings were trans- remained for 9 d (period for development from egg to ferred to new cages. During evaluation, the branches pupa), according to Rosa et al. (2012). After this period, with parasitized nymphs (mummified) were cut off and the seedlings with T. radiata pupae were sprayed with transferred to Petri dishes (9 cm in diameter by 0.7 cm the insecticides, using a Guarany backpack sprayer in height) containing moistened filter paper, to assess (Guarany Indu´stria e Come´rcio Ltda., Sa˜o Paulo, Bra- the number of ectoparasitoid adults emerged (emer- zil) equipped with a TXVS-4 conical nozzle (Teejet gence rate). The adults obtained in each treatment Technologies, Sa˜o Paulo, Brazil) to the runoff point, were counted, separated by sex (sex ratio), and placed and each seedling was used as the experimental unit. in individual glass tubes (2.5 cm in diameter by 8.5 cm Distilled water was used as a control treatment. For in length) with a drop of honey, to evaluate the longev- each treatment, five repetitions with five seedlings per ity of the insects. The insects (females and males) were repetitions were used. observed every 24 h until they died. After the spraying treatments, the experimental units Based on the mortality data (acute toxicity), on the were kept in a climate-controlled room for 3 h to allow parasitism (parasitism rate), and on the number of the residues to dry. Afterward, branches with T. radiata emerged parasitoids (emergence rate) of T. radiata,the pupae were cut off from each seedling, and placed on total effect for each acaricide was estimated using the moistened filter paper in Petri dishes (9 cm in diame- formula Eð%Þ¼100 ð100 MÞEr, proposed by ter) to evaluate the acute toxicity of the acaricides. The Van de Veire et al. (2002), where: M ¼ mortality cor- number of emerged adults in each experimental unit rected according to the formula of Abbott (1925) and was evaluated every 24 h for 8 d. The mortality of each Er ¼ sublethal effects, calculated as Er ¼ R1 R2, treatment was calculated based on the number of where R1 ¼ ratio of the mean number of nymphs para- pupae (parasitized nymphs) and the number of adults sitized (mummified) by female in the acaricide treat- emerged in each experimental unit. The acute toxicity ment and the control, and R2 ¼ ratio of the mean (reduction in the emergence rate of the F0 generation) number of emerged parasitoids (emergence rate) in the of each acaricide was calculated based on mean mortal- acaricide treatment and the control. Based on the total ity observed in each acaricide treatment and the con- effect (E), each acaricide was classified according to trol. Adults surviving in each experimental unit were the criteria established by the IOBC/WPRS for counted, separated by sex (sex ratio), and placed in extended laboratory tests (Van de Veire et al. 2002): glasscages(90by50by50cm3) containing seedlings class 1 ¼ harmless (E < 25%), class 2 ¼ slightly harmful infested with D. citri nymphs of fourth and fifth instars (25% E 50%), class 3 ¼ moderately harmful to evaluate the effects of the acaricides on the parasit- (51% E 75%), and class 4 ¼ harmful (E > 75%). ism rate (F0 generation) and emergence rate (F1 gener- Bioassay 3: Acute Toxicity and Sublethal Effects ation) of the ectoparasitoid. For this, the D. citri of Acaricides Applied to T. radiata Pupae. To eval- nymphs were exposed to T. radiata females for parasit- uate the acute toxicity and sublethal effects of acari- ism for 48 h. After this period, surviving T. radiata cides on pupae of T. radiata, acaricides that were adults were transferred to individual glass tubes (2.5 cm considered harmless (class 1) to ectoparasitoid adults in in diameter by 8.5 cm in length) to evaluate the longev- the assay of acute toxicity (bioassay 1) were used. Our ity. During this period, the adults were fed with drops decision to use such acaricides was due to its mecha- of honey as described for bioassay 1. nism of action (see Table 1). Acaricides that act as mite The parasitism rate (F0 generation) of T. radiata was growth inhibitors and energy production can reduce determined based on the number of parasitized (mum- the emergence, survival, and parasitism rates of off- mified) D. citri nymphs in each experimental unit, 9 d spring when these products are applied on immature after the T. radiata adults were removed from the cages. stages of the parasitoids, due to ingestion and contact The emergence rate (F1 generation) of the ectoparasi- with the acaricide residues at the time of, or after, toid was determined based on the number of T. radiata emergence. In this context, we decided to evaluate the adults emerged in each experimental unit 15 d after the possible effects of these acaricides on pupae of T. radi- removal of T. radiata adults from the cages. Based on ata to verify the compatibility of these products with the acute toxicity (reduction in the emergence rate of the ectoparasitoid before recommending its use in the F0 generation) and sublethal effects (parasitism rate 2015 LIRA ET AL.: EFFECT OF 16 ACARICIDES ON Tamarixia radiata 5

in the F0 generation, and emergence rate in the F1 gen- Table 2. Mean number of dead insects and standard error eration) on T. radiata, the total effect was estimated for mean (N 6 SEM) and corrected mortality (%) of T. radiata exposed to residual contact acaricides each acaricide, using the same procedure and criteria for analysis described in bioassay 2. Treatment Dosage used Acute toxicity IOBC/WPRS Bioassay 4: Duration of Harmful Effects of (g a. i. liter1) a classb Acaricides on T. radiata Adults. The duration of the N 6 SEM % harmful effects of the acaricides was assessed on Control – 0.1 6 0.22 – – T. radiata adults, using the acaricides that were consid- Abamectin 0.005 2.8 6 0.83 27.3 2 Acrinathrin 0.005 8.0 6 0.64 79.8 4 ered slightly harmful or harmful to ectoparasitoid Bifenthrin 0.020 10.0 6 0.00 100.0 4 adults in the bioassay of acute toxicity (bioassay 1), Carbosulfan 0.350 10.0 6 0.00 100.0 4 using the method proposed by Brunner et al. (2001). Dicofol 0.370 2.4 6 0.92 23.2 1 Five Valencia sweet orange seedlings 80 cm tall and Diflubenzuron 0.120 0.4 6 0.44 3.0 1 Etoxazole 0.021 0.6 6 0.23 5.0 1 with 20 mature leaves, for each treatment, were Fenbutatin oxide 0.400 4.0 6 1.56 39.4 2 sprayed with acaricides and distilled water (control Fenpropathrin 0.150 9.8 6 0.23 97.8 4 treatment) until the runoff point, using a Guarany Fenpyroximate 0.050 3.8 6 0.75 37.4 2 backpack sprayer equipped with a TXVS-4 conical noz- Flufenoxuron 0.050 1.5 6 0.87 14.1 1 Hexythiazox 0.015 0.2 6 0.22 1.0 1 zle. After the treatments were applied, the seedlings Propargite 0.720 5.0 6 0.64 49.5 2 were placed in a greenhouse. At 1, 3, 7, 10, 17, 24, and Pyridaben 0.150 2.5 6 1.0 24.2 1 30 d after the treatments, one treated leaf of each plant Spirodiclofen 0.060 2.2 6 0,8 21.2 1 was randomly removed, brought to the laboratory, and Sulfur 4.000 5.0 6 1.6 49.5 2 discs (3.3 cm in diameter) were cut for use in preparing a Field dosage recommended for the control of mites pest in citrus. the experimental units as described in bioassay 1. b IOBC/WPRS class based on parasitoid mortality: class 1 ¼ Then, 10 T. radiata adults up to 48 h old were anesthe- harmless (M < 25%), class 2 ¼ slightly harmful (25% M 50%), class 3 ¼ moderately harmful (51% M 75%), and class 4 ¼ harmful tized with CO2 for 5 s and transferred into each experi- (M > 75%). mental unit. The experimental units were sealed with voile fabric and placed in a climate-controlled room. For each treatment, five repetitions were used acaricides caused mortality rates from 29.1 to 42.9% (n ¼ 50). and were considered slightly harmful (class 2). Dicofol, The mortality of insects was recorded after 24 h of diflubenzuron, etoxazole, flufenoxuron, hexythiazox, exposure to the residues. Insects that did not react to pyridaben, and spirodiclofen caused <20% mortality the touch of a fine brush were considered dead. The and were considered harmless (Table 2). Despite their mortality data for each acaricide and dates of assess- low acute toxicity, the acaricide diflubenzuron, etoxa- ment were corrected by the formula of Abbott (1925). zole, flufenoxuron, and spirodiclofen reduced the para- The products that reduced adult survival by <25% sitism rate and the number of emerged insects of the compared to the control treatment (distilled water) F1 generation, and were therefore considered slightly were classified according to the persistence scale pro- harmful (class 2) to T. radiata (Table 3). Diflubenzuron, posed by the IOBC/WPRS: class 1: short life (<5d), etoxazole, and flufenoxuron decreased the emergence class 2: slightly persistent (5–15 d), class 3: moderately of females (sex ratio) in the F1 generation compared to persistent (16–30 d), and class 4: persistent (>30 d; Van the other treatments. The longevity was lower in adults de Veire et al. 2002). exposed to residues of diflubenzuron, flufenoxuron, Data Analysis. Generalized linear models (Nelder and spirodiclofen, than in the control treatment and Wedderburn 1972) with Gaussian distributions (Table 3). Dicofol and pyridaben did not affect the par- were used for the data analysis of the longevity of asitism rate, emergence rate, sex ratio, or longevity, and insects. The quality of the adjustment was verified with were considered harmless (class 1) to adults of the a half-normal plot with simulated envelope (Hinde and ectoparasitoid (Table 3). Deme´trio 1998). Multiple comparisons by the Tukey– Acute Toxicity and Sublethal Effects of Kramer test (P 0.05) using the “glht” function of the Acaricides on T. radiata Pupae. The application of “DTK” package, with adjustment of the P values, were acaricides to T. radiata pupae indicated that difluben- used to compare the means of different treatments. Sex zuron, etoxazole, flufenoxuron, and hexythiazox did not ratio data for treatments were compared using the Chi- affect the emergence rate of adults or the sex ratio and square test (P 0.05). All analyses were performed longevity of T. radiata in the F0 generation, but did with the statistics software R version 2.15.1 (R Devel- reduce the parasitism rate in the F0 generation and opment Core Team 2012). emergence rate in the F1 generation, and were therefore considered slightly harmful (class 2) to T. radiata (Table 4). Dicofol and pyridaben caused mortality rates Results of 24.8 and 28.9%, respectively, but led to increases in Acute Toxicity and Sublethal Effects of the parasitism rate and did not affect the emergence Acaricides on T. radiata Adults. Residues of acrina- rate, sex ratio, or longevity of the ectoparasitoid, and thrin, bifenthrin, carbosulfan, and fenpropathrin were were considered innocuous (class 1) to T. radiata.Spi- harmful (class 4) to T. radiata adults (mortality greater rodiclofen did not significantly influence any of the var- than 77% after 24 h of exposure; Table 2). Abamectin, iables evaluated, and was also classified as harmless fenbutatin oxide, fenpyroximate, propargite, and sulfur (class 1) to T. radiata (Table 4). 6JOURNAL OF ECONOMIC ENTOMOLOGY

Table 3. Sublethal and total effects of acaricides applied to adults of T. radiata

a b c d f g Treatment Dosage used M (%) R1 R2 E (%) IOBC/WPRS Sex ratio Longevity (d) (g a. i. liter1) classe Control – 4.0 6.30 6.25 – – 0.74 6 0.07a 6.2 6 0.21a Dicofol 0.370 19.4 8.06 5.56 3.6 1 0.79 6 0.02a 4.7 6 0.48b Diflubenzuron 0.120 4.0 6.49 4.06 37.0 2 0.59 6 0.03b 4.0 6 0.18b Etoxazole 0.021 4.0 5.42 3.75 48.4 2 0.64 6 0.01b 6.3 6 0.58a Flufenoxuron 0.050 7.4 6.24 4.38 33.1 2 0.61 6 0.04b 4.3 6 0.13b Hexythiazox 0.015 14.0 0.91 5.31 30.4 2 0.77 6 0.05a 6.2 6 0.78a Pyridaben 0.150 24.0 7.94 5.38 13.3 1 0.87 6 0.01a 6.0 6 0.81a Spirodiclofen 0.060 23.9 7.62 4.25 34.1 2 0.80 6 0.03a 4.5 6 0.68b v2 ¼ 0.02178 F ¼ 3.367 df ¼ 7df¼ 7, 32 P ¼ 0.010 P ¼ 0.0464 a M—Mortality corrected by formula of Abbott (1925). b R1—Effect on parasitism (mean number of parasitized nymphs). c R2—Effect on emergence (mean number of adults emerged in F1 generation). d E—Total effect of acaricides calculated by formula of Van de Veire et al. (2002). e IOBC/WPRS class based on parasitoid mortality: class 1 ¼ harmless (M < 25%), class 2 ¼ slightly harmful (25% M 50%), class 3 ¼ moderately harmful (51% M 75%), and class 4 ¼ harmful (M > 75%). f Means followed by the same letter in a column do not differ significantly by Chi-square test (P 0.05). g Means followed by the same letter in a column do not differ significantly (GLM with Gaussian distribution, followed by post hoc Tukey test; P 0.05).

Table 4. Total effects of acaricides applied to pupae of T. radiata

a b c d Treatment Dosage used M (%) R1 R2 E (%) IOBC/WPRS Sex ratio Longevity (d) 1 e f g (g a. i. liter ) class (F1 generation) (F1 generation) Control – 4.0 6.82 6.80 – – 0.69 6 0.06a 7.9 6 1.61a Dicofol 0.370 24.8 8.32 7.48 15.2 1 0.82 6 0.14a 6.7 6 1.32a Diflubenzuron 0.120 13.2 6.82 4.08 45.5 2 0.75 6 0.19a 7.8 6 1.17a Etoxazole 0.021 6.0 6.17 4.76 40.8 2 0.80 6 0.06a 7.6 6 1.61a Flufenoxuron 0.050 12.6 6.34 4.96 40.5 2 0.70 6 0.14a 7.3 6 1.13a Hexythiazox 0.015 19.7 6.14 5.54 38.6 2 0.79 6 0.10a 6.0 6 1.36a Pyridaben 0.150 28.9 7.18 7.48 13.3 1 0.77 6 0.05a 6.5 6 0.51a Spirodiclofen 0.060 9.2 7.20 6.12 9.6 1 0.76 6 0.08a 8.6 6 1.31a v2 ¼ 0.222 F ¼ 0.468 df ¼ 7df¼ 7, 32 P ¼ 0.236 P ¼ 0.826 a M—Effect on emergence rate (F0 generation) calculated by formula of Abbott (1925). b R1—Effect on parasitism (mean number of parasitized nymphs in F0 generation). c R2—Effect on emergence (mean number of adults emerged in F1 generation). d E—Total effect of acaricides calculated by formula of Van de Veire et al. (2002). e IOBC/WPRS class based on parasitoid mortality: class 1 ¼ harmless (M < 25%), class 2 ¼ slightly harmful (25% M 50%), class 3 ¼ moderately harmful (51% M 75%), and class 4 ¼ harmful (M > 75%). f Means followed by the same letter in a column do not differ significantly by Chi-square test (P 0.05). g Means followed by the same letter in a column do not differ significantly (GLM with Gaussian distribution, followed by post hoc Tukey test; P 0.05).

Duration of the Harmful Effect of Acaricides on Discussion T. radiata Adults. The application of acaricides that Acute toxicity and sublethal effects of 16 acaricides were considered slightly harmful or harmful to adults recommended for the control of pest mites in citrus of T. radiata on citrus seedlings in the greenhouse indi- were evaluated on the ectoparasitoid T. radiata.The cated that carbosulfan maintained its harmful activity acaricides showed different toxicity levels according to (25% mortality) for >30 d after spraying (DAS), and the chemical group used in the bioassays. Acrinathrin, was therefore considered persistent (>30 d, class 4) bifenthrin, and fenpropathrin pyrethroids and carbosul- according to the IOBC/WPRS study group criteria fan carbamate showed the highest acute toxicity to (Table 5). However, acrinathrin, bifenthrin, fenpropa- T. radiata adults, with >77% mortality 24 h after expo- thrin, fenpyroximate, propargite, and sulfur caused sure. These results are similar to those obtained by mortality exceeding 25% until 24 DAS, and were con- Hall and Nguyen (2010), who reported mortality rates sidered moderately persistent (16–30 d, class 3). Aba- above 91.3% for T. radiata adults, 24 h after direct mectin was slightly persistent (5–15 d, class 2) and spraying or exposed to dry residues of fenpropathrin. fenbutatin oxide was evaluated as short life (<5d,class In addition to their effects on T. radiata, bifenthrin and 1; Table 5). fenpropathrin also caused high mortality to adults of 2015 LIRA ET AL.: EFFECT OF 16 ACARICIDES ON Tamarixia radiata 7

Table 5. Mortality (%) of T. radiata adults 1, 3, 7, 10, 17, 24, and 30 d after spraying of the acaricides

Treatment Dosage Mortality (%)—days after spraying (DAS) IOBC/ used WPRS (g a. i. liter1) 1 3 7 10172430classa Control – 6.0 6 0.43 2.0 6 0.32 4.0 6 0.44 0.0 6 0.00 2.0 6 0.32 2.0 6 0.32 0.0 6 0.00 – Abamectin 0.005 31.7 6 3.75 37.8 6 7.43 30.5 6 7.13 29.5 6 6.13 10.5 6 4.73 5.2 6 4.73 6.0 6 0.40 2 Acrinathrin 0.005 87.5 6 3.27 73.5 6 4.44 68.3 6 9.42 38.1 6 3.33 15.8 6 7.21 8.3 6 4.55 4.9 6 0.82 3 Bifenthrin 0.020 100.0 6 0.00 100.0 6 0.00 87.5 6 2.17 70.0 6 4.64 29.3 6 4.25 22.2 6 5.43 18.8 6 9.93 3 Carbosulfan 0.350 100.0 6 0.00 100.0 6 0.00 92.5 6 3.24 67.5 6 8.37 43.2 6 1.37 37.0 6 3.57 31.1 6 5.37 4 Fenbutatin oxide 0.400 33.5 6 4.93 12.6 6 6.23 17.5 6 5.35 12.4 6 7.75 18.9 6 2.62 4.0 6 0.58 3.8 6 2.94 1 Fenpropathrin 0.050 93.6 6 4.38 97.8 6 2.24 87.5 6 6.07 60.5 6 6.91 37.4 6 5.68 14.8 6 8.22 38.3 6 4.83 3 Fenpyroximate 0.150 40.4 6 6.46 49.6 6 8.76 43.0 6 3.69 36.0 6 4.65 27.4 6 4.46 9.6 6 6.83 18.3 6 8.47 3 Propargite 0.720 50.2 6 7.92 52.2 6 8.14 39.5 6 2.71 40.5 6 2.48 33.1 6 4.12 8.3 6 4.56 48.9 6 3.86 3 Sulfur 4.000 95.7 6 4.30 95.7 6 4.45 88.1 6 3.56 88.1 6 6.56 58.9 6 6.94 19.1 6 1.24 21.3 6 7.23 3 a IOBC/WPRS class based on the duration of the harmful effect of acaricides: class 1 ¼ short life (<5 d), class 2 ¼ slightly persistent (5–15 d), class 3 ¼ moderately persistent (16–30 d), and class 4 ¼ persistent (>30 d).

Aphytis melinus Debach (Hymenoptera: Aphelinidae) citrus-producing regions of Brazil, this period coincides (Michaud and Grant 2003), Encarsia formosa (Gahan) with the dry months of the year (from mid-autumn to (Hymenoptera: Aphelinidae), and Eretmocerus eremi- mid-spring), when the vegetative growth rate of citrus cus Rose & Zolnerowich (Hymenoptera: Aphelinidae) plants (sprouts) slows significantly, substantially (Prabhaker et al. 2007). However, only slightly acute decreasing infestation levels of D. citri nymphs (Yama- toxicity (30–79% mortality) was observed in adults of moto et al. 2001) and the population of T. radiata Chrysocharis pentheus (Kamijo) and Sympiesis stria- adults in orchards (Paiva and Parra 2012). It should be tipes Ashmead (Hymenoptera: Eulophidae) (Mafi and emphasized, however, that this period also coincides Ohbayash 2006)andTrichogramma chilonis Ishii and with the initial infestation of B. phoenicis (a main pest Trichogramma brasiliensis (Ashmead) (Hymenoptera: mite of citrus) and with the highest population levels of Trichogrammatidae) (Shankarganesh et al. 2013) after tetranychid mites (especially P. citri and E. banksi)that 24 h of exposure to residues of bifenthrin and carbosul- cause considerable damage in plants and, in most cases, fan, respectively. The high acute toxicity of these acari- require chemical intervention for their control (Oliveira cides is attributed to high absorption of the products and Pattaro 2008). Therefore, during this period, the by insects during the walking on the surfaces treated use of these acaricides for management of these targets with acaricides, the rapid penetration capacity of prod- will not cause negative impacts on T. radiata adults. ucts in the integument, and the low capacity of detoxifi- The exposure of T. radiata adults to residues of aba- cation of active ingredients by enzymes oxidases and mectin, fenbutatin oxide, fenpyroximate, propargite, hydrolases (Siegfried 1993). Therefore, our results indi- and sulfur, at recommended concentrations for the cate that the use of these acaricides in citrus orchards control of B. phoenicis and P. oleivora in citrus to control pest mites can severely affect the survival of orchards (Table 1), revealed that these acaricides T. radiata, and inhibit the action of this ectoparasitoid caused low acute toxicity (<42.9% mortality) to T. radi- in controlling D. citri. ata adults. However, our results were different from In addition, acrinathrin, bifenthrin, and fenpropa- those obtained by Hall and Nguyen (2010), who found thrin pyrethroids were moderately persistent (25% high mortality (>79%) of T. radiata adults subjected to mortality between 17 and 24 d after the products were direct spray or exposed to residual contact of 0.016 g sprayed), whereas the carbosulfan carbamate retained liter1 abamectin. Likewise, Cocco and Hoy (2008) its harmful activity for >30 d, and was considered per- observed high acute toxicity (93.1% mortality) to sistent for adults of T. radiata.Similarly,Prabhaker T. radiata adults sprayed with abamectin in the lowest et al. (2007) found that, under laboratory conditions, label rate (0.40 g liter1). In addition to T. radiata, the residual effect of fenpropathrin persisted longer others parasitoids species demonstrated high suscepti- than 21 d for adults of A. melinus, while in the field, bility to abamectin. Luna-Cruz et al. (2011) and Liu Nguyen and Hall (2010) foundupto64.4%reduction et al. (2012) evaluating the acute toxicity of abamectin in the population levels of adult T. radiata 22 d after at concentrations of 0.75 and 0.0112 g liter1, respec- the product was sprayed. The long residual period may tively, on adults of Tamarixia triozae (Burks) (Hyme- be explained by high adherence capacity of these prod- noptera: Eulophidae), found that these acaricides ucts in citrus leaves and by lower degradation by envi- caused high mortality of the parasitoid after 24 h of ronmental factors (Yadav et al. 2003). Thus, our results exposure to residues. Likewise, Hernańdez et al. (2011) suggest that the successive use of these products can observed high mortality of Ganaspidium igrimanus compromise biological control of D. citri exercised by (Kieffer) (Hymenoptera: Figitidae) adults when it was ectoparasitoid T. radiata. exposed to residual contact of 0.1123 g liter1 abamec- Despite their high acute toxicity and long residual tin, whereas Vanaclocha et al. (2013) found high sus- period, these acaricides may be used in periods of low ceptibility of A. melinus adults kept for 24 h on leaves mobility and density of adults of T. radiata,toexploit of Clementine [Citrus clementine Hort. (Rutaceae)] the ecological selectivity of these products. In the main treated with 0.40 g liter1 of acaricide. The lower acute 8JOURNAL OF ECONOMIC ENTOMOLOGY toxicity observed in this study is attributed to lower (Lepidoptera: Gracillariidae)], which infests citrus concentration of abamectin used in bioassays. In addi- plants during the vegetative growth phase (Paiva 2011). tion, not adding oil at the time of product application Fenbutatin oxide, fenpyroximate, propargite, and sulfur may have intensified the degradation process of aba- are recommended for management of Polyphagotarso- mectin present in the foliar surface by environmental nemus latus (Bancks) (Acari: Tarsonemidae), which also factors (Vanaclocha et al. 2013). infests plants during vegetative growth. This period Acute toxicity similar to that found in the present also coincides with higher infestation levels of D. citri study was observed in adults of Trichogramma cacoe- nymphs in citrus plants, as well as of its ectoparasitoid ciae Marchal (Hymenoptera: Trichogrammatidae) T. radiata. Therefore, these acaricides should be used (Saber 2011)andE. eremicus (Sugiyama et al. 2011) with caution in order to avoid compromising the effi- exposed to residues of fenpyroximate and sulfur, cacy of this ectoparasitoid in biological control of respectively. On the other hand, fenpyroximate was D. citri in citrus orchards. considered innocuous to T. triozae adults (Liu et al. Dicofol, diflubenzuron, etoxazole, flufenoxuron, hex- 2012); propargite was safe for E. formosa and A. meli- ythiazox, and spirodiclofen did not cause significant nus adults (Gholamzadeh et al. 2012, Vanaclocha et al. mortality (<25%) to T. radiata adults. Hall and Nguyen 2013); and fenbutatin oxide did not cause mortality of (2010) found similar results for T. radiata adults Aphidius colemani Viereck (Hymenoptera: Braconidae) sprayed directly or exposed to dry residues of difluben- adults (Urbaneja et al. 2008). Based on the low acute zuron. Diflubenzuron and flufenoxuron were consid- toxicity, our results indicate that the use of these acari- ered nontoxic to adults of Telenomus remus Nixon cides to control pest mites can reduce the population (Hymenoptera: Scelionidae) (Carmo et al. 2010), and of T. radiata adults and the effectiveness of ectoparasi- flufenoxuron and lufenuron did not cause mortality of toid in the control of D. citri in citrus orchards. adults of Eretmocerus mundus (Mercet) (Hymenop- In addition, fenbutatin oxide, abamectin, fenpyroxi- tera: Aphelinidae), E. eremicus,andE. formosa mate, propargite, and sulfur retained their harmful (Sugiyama et al. 2011). Likewise, Vanaclocha et al. activity for up to 1, 10, 17, 24, and 24 d after the prod- (2013) reported that the use of etoxazole and hexythia- ucts were sprayed, respectively. Fenbutatin oxide also zox was compatible with the parasitoid A. melinus, showed low residual persistence (short life) to adults of demonstrating that these acaricides do not affect the A. melinus parasitoid (<10% mortality until 6 d after survival of adult parasitoids. The low mortality of adults spraying; Morse et al. 1987) and for females of preda- is attributed to action mechanism of products. Diflu- tory mites Iphiseiodes zuluagai Denmark and Muma benzuron and flufenoxuron act by inhibition of chitin (Acari: Phytoseiidae) (<30% mortality, 1 d after spray- biosynthesis at the time of ecdysis (Willrich and Boe- ing) and Euseius alatus DeLeon (Acari: Phytoseiidae) thel 2001), whereas etoxazole and hexythiazox inhibit (<30% mortality until 5 d after spraying) of product the growth of the arthropods by a mechanism similar (Reis and Sousa 2001), demonstrating that this acari- to benzoylphenylureas (Demaeght et al. 2014). Thus, cide caused less impact on the biological control agents these products present high activity while applied on present in citrus orchards. The low residual persistence immature stage of insects, but did not affect the sur- of fenbutatin oxide on T. radiata adults is a desirable vival of parasitoid adults. feature and important for integrated pest management Although they did not cause adult mortality, residues programs, because it allows the establishment of ecto- of diflubenzuron, etoxazole, and flufenoxuron led to parasitoid in the production area in a short period of lower longevity (except etoxazole) and parasitism rates time after application. On the other hand, abamectin, in T. radiata females compared to the control. These fenpyroximate, propargite, and sulfur demonstrated a acaricides also reduced the emergence rate and sex lower compatibility with the ectoparasitoid T. radiata. ratio (proportion of females) in the offspring (F1 prog- However, these results were obtained in laboratory, eny). Similarly, Schneider et al. (2004) observed lower where the insects were continually exposed to residues longevity and parasitism rate in Hyposoter didymator of acaricides (maximum harmful potential); probably in (Thunberg) (Hymenoptera: Ichneumonidae) females field conditions, the acute toxicities and the residual exposed to residues of diflubenzuron, and Tabozada effects of these products will be lower than those et al. (2014) found reductions in parasitism ability, observed here. This hypothesis is supported by the emergence rate, sex ratio, and longevity of Trichog- study of Nguyen and Hall (2010), who found a residual ramma evanescens (Westw.) (Hymenoptera: Trichog- period of abamectin and sulfur of 8 d, whereas Morse rammatidae) and Bracon brevicornis Wesmael et al. (1987) and Vanaclocha et al. (2013) reported a (Hymenoptera: Braconidae) treated with flufenoxuron. mortality of A. melinus adults of 10 and 38.3% at 6 and The lowest emergence rate in the F1 generation may 7 d after spraying of abamectin, respectively. Therefore, be attributed to the mortality of parasitoid larvae within in areas with high incidences of pest mites, where D. citri nymphs, caused by malformation of the parasi- releases of T. radiata will be conducted to control toid integument (lower chitin biosynthesis) before its D. citri, our results indicate that releases can be per- emergence (Nauen and Smagghe 2006, Schneider formed safely 1, 10, 17, 24, and 24 d after spraying of et al. 2008). The reduction in longevity and the emer- fenbutatin oxide, abamectin, fenpyroximate, propargite, gence rate and mainly in the sex ratio of offspring, fur- and sulfur, respectively. Notably, however, abamectin is ther reduces the effectiveness of the ectoparasitoid in one of the most-used active ingredients in the control biological control of D. citri, because only the female of citrus leafminer [Phyllocnistis citrella Station wasps parasitize this psyllid. In addition to their 2015 LIRA ET AL.: EFFECT OF 16 ACARICIDES ON Tamarixia radiata 9 sublethal effects on adults, these acaricides also Lessons from huanglongbing management in SaõPaulostate, reduced the emergence rate and parasitism rate of T. Brazil. J. Plant Pathol. 92: 285–302. radiata females originated from pupae treated with Bove´, J. M. 2006. Huanglongbing: A destructive, newly-emerg- acaricides. Although the pupal stage is considered to be ing, century-old disease of citrus. J. Plant Pathol. 88: 7–37. more tolerant than adults to xenobiotic agents, our Brunner, J. F., J. E. Dunley, M. D. Doerr, and A. H. Beers. 2001. Effect of pesticides on Colpoclypeus florus (Hymenop- results showed that spraying these products may affect tera: Eulophidae) and Trichogramma platneri (Hymenoptera: the biological and behavioral parameters of the ectopar- Trichogrammatidae), parasitoids of leafrollers in Washington. asitoid. The harmful effects of acaricides sprayed on T. J. Econ. Entomol. 94: 1075–1084. radiata pupae are attributed to their penetration ability Carmo, E. L., A. F. Bueno, and R.C.O. F. Bueno. 2010. Pes- and to direct contact with the acaricide residues when ticide selectivity for the insect egg parasitoid Telenomus the parasitoid emerges. These hypotheses are sup- remus. Biocontrol 55: 455–464. ported by the studies of Schneider et al. (2003, 2004), Chen,X.,andP.A.Stansly.2014.Effect of holding diet on egg who found that diflubenzuron easily penetrated the formation of Tamarixia radiata (Hymenoptera: Eulophidae), integument of H. didymator when applied on the para- parasitoid of Diaphorina citri (Hemiptera: Psylloidae). Fla. Entomol. 97: 491–495. sitoid pupae. However, future studies should be con- Cocco, A., and M. A., Hoy 2008. Toxicity of organosilicone ad- ducted to elucidate the main forms of contamination of juvants and selected pesticides to the Asian citrus psyllid T. radiata pupae by active ingredients of acaricides. (Hemiptera: Psyllidae) and its parasitoid Tamarixia radiata Of the acaricides evaluated, only dicofol and pyrida- Hymenoptera: Eulophidae). Fla. Entomol. 91: 610–620. ben were harmless to T. radiata.However,Hall and Demaeght, P., E. J. Osborne, J. Odman-Naresh, M. Grbic, Nguyen (2010) reported that the use of pyridaben in R.Nauen,H.Merzendorfer,R.M.Clark,andT.V. the concentration of 0.375 g liter1 (2.5 times higher Leeuwen. 2014. High resolution genetic mapping uncovers than in this study) caused low mortality of T. radiata chitin synthase-1 as the target-site of the structurally diverse adults exposed to contact with the product residues, mite growth inhibitors clofentezine, hexythiazox and etoxazole in Tetranychus urticae. Insect Biochem. Mol. Biol. 51: demonstrating that the acaricide is compatible with 52–61. IPM programs. Likewise, Yamamoto and Bassanezzi Desneux, N., A. Decourtye, and J. M. Delpuech. 2007. The (2003) reported that dicofol was safe for Ageniaspis cit- sublethal effects of pesticides on beneficial arthropods. Annu. ricola (Hymenoptera: Encyrtidae) adults, an important Rev. Entomol. 52: 81–106. parasitoid of P. citrella. Previous studies have shown (FUNDECITRUS) 2013. 7,2 milho˜esdeplantassaõ arrancadas that dicofol and pyridaben were harmless or only por causa do greening em 2012. (http://www.fundecitrus.com. slightly harmful to Azya luteipes Mulsant, Coccidophi- br/Noticias/27.03.13—7,2-milhoes-de-plantas-sao-arranca lus citricola Bre`thes, Cycloneda sanguinea (L.), and das-por-causa-do-greening-em-2012,333). Pentilia egena Mulsant (Coleoptera: Coccinellidae), Gholamzadeh, M., M. Ghadamyari, L. Salehi, and V. Hosei- ninaveh. 2012. Effects of amitraz, buprofezin and propar- and lacewings (Neuroptera: Chrysopidae), important gite on some fitness parameters of the parasitoid Encarsia predators of arthropod pests in citrus orchards (Yama- formosa (Hym.: Aphelinidae), using life table and IOBC moto and Bassanezzi 2003). These results indicate that methods. J. Entomol. Soc. Iran 31: 1–14. these acaricides are safe and can be used at times of Go´ mez-Torres, M. L. 2009. Estudos bioecolo´gicos de Tamar- high population density or in conjunction with inunda- ixia radiata (Waterston, 1922) (Hymenoptera: Eulophidae) tive releases of T. radiata not affecting the survival and para o controle de Diaphorina citri Kuwayama, 1907 (Hemi- effectiveness of the ectoparasitoid for biological control ptera: Psyllidae). Ph.D. thesis, “Luiz de Queiroz” College of of D. citri. Agriculture/University of Saõ Paulo (ESALQ/USP), Piraci- caba, SP. Go´mez-Torres,M.L.G.,D.E.Nava,S.Gravena,V.A. Costa, and J.R.P. Parra. 2006. Registro de Tamarixia radiata (Waterston) (Hymenoptera: Eulophidae) em Diaphorina Acknowledgments citri Kuwayama (Hemiptera: Psyllidae) em Saõ Paulo, Brasil. Rev. Agric. 81: 112–117. We thank the Brazilian Federal Agency for the Support Go´ mez-Torres, M. L., D. E. Nava, and J.R.P. 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