Insect Growth Regulator Fenoxycarb Research Articles

Development and characterization of Spodoptera mauritia ovarian primary cell culture and evaluation of fenoxycarb toxicity.

The rice swarming caterpillar, Spodoptera mauritia (Biosduval, 1833) (Lepidoptera: Noctuidae), has been considered a severe pest and has caused the extreme loss of rice paddies in the nursery stages. The massive reproductive efficiency of pests results from balanced endocrine functioning. Insect treatments with exogenous either juvenile hormones (JH) or juvenile hormone analogues (JHAs) during low endogenous JH titers disrupt metamorphosis andovarian development. Hence, it was thought worthwhile to develop a primary ovarian cell culture of Spodoptera mauritia to study the biological efficiency of JHA, fenoxycarb, at the cellular level. The study envisioned the cell characteristics and growth properties and found that most cells were spherical. Spindle-shaped cells were also present during the initial stage of active cell division. The majority of the cells grew attached to the bottom of the culture plates, and a few grew in suspension. The cell doubling time was 144 ± 14h. The growth curve exhibited a sigmoid pattern of cell proliferation at first, followed by a declining phase. These properties indicate that the primary ovarian culture is a suitable candidate for developing a novel cell line of S. mauritia. The preliminary study investigated the cytotoxicity of the insect growth regulator fenoxycarb in ovarian, primary cell culture. Incubated cells showed noticeable morphological changes, including cell shrinkage and apoptotic body formation; inhibition of cellular proliferation; and apoptosis-inducing effects in ovarian, primary cell culture of S. mauritia. Evidently, from these results, it is clear that juvenile hormone mimics can react in a meaningful way to control the rice pest S. mauritia by inducing cellular apoptosis in ovarian cells, which may cause adult females to become sterile.

Efficacy of biorational insecticides against Bemisia tabaci (Genn.) and their selectivity for its parasitoid Encarsia formosa Gahan on Bt cotton

The toxicity of seven biorational insecticides [five insect growth regulators (Buprofezin, Fenoxycarb, Pyriproxyfen, Methoxyfenozide, and Tebufenozide) and two oil-extracts of neem and bitter gourd seeds] against Bemisia tabaci and their selectivity for its parasitoid, Encarsia formosa were evaluated in laboratory and field conditions for 2 years (2018–2019) in Pakistan. Toxicity results demonstrate that Pyriproxyfen, Buprofezin, and Fenoxycarb proved to be effective (80–91% mortality and 66.3–84.2% population-reduction) against B. tabaci followed by Methoxyfenozide, Tebufenozide (50–75% mortality and 47.8–52.4% population-reduction), and then oil-extracts of neem and bitter gourd (25–50% mortality and 36.5–39.8% population-reduction) in the laboratory [72 h post-application exposure interval (PAEI)] and field trails (168 h PAEI), respectively. All tested biorationals, except Methoxyfenozide [(slightly-harmful/Class-II), i.e., causing mortality of parasitoids between a range of 25–50%] and Tebufenozide [(moderately-harmful/Class-III), i.e., causing mortality of parasitoids between the ranges of 51–75%], proved harmless/Class-I biorationals at PAEI of 7-days in the field (parasitism-reduction < 25%) and 3-days in the lab (effect < 30%). In laboratory bioassays, exposure of parasitized-pseudopupae and adult-parasitoids to neem and bitter gourd oils demonstrated that these compounds proved harmless/Class-I biorationals (< 30% mortality). Alternatively, Pyriproxyfen, Buprofezin, Fenoxycarb, Methoxyfenozide, and Tebufenozide were slightly-harmful biorationals (30–79% mortality) against the respective stages of E. formosa. We conclude that most of the tested biorationals proved harmless or slightly harmful to E. formosa, except tebufenozide after PAEI of 7-days (168 h) in the field and, therefore, may be used strategically in Integrated Pest Management (IPM) of B. tabaci.

Fenoxycarb exposure disrupted the reproductive success of the amphipod Gammarus fossarum with limited effects on the lipid profile.

Insect growth regulator insecticides mimic the action of hormones on the growth and development of insect pests. However, they can affect the development of non-target arthropods. In the present study, we tested the effects of the growth regulator insecticide fenoxycarb on several endpoints in the freshwater crustacean Gammarus fossarum (Amphipoda). Females carrying embryos in their open brood pouch were exposed to 50 μg L-1 fenoxycarb throughout the entire oogenesis (i.e. 21 days). After exposure, newborn individuals from exposed embryos were removed from the maternal open brood pouch for lipidomic analysis, while males were added to assess the reproductive success. After fertilization, the lipid profile, energy reserve content (lipids, proteins and glycogen), and activity of phenoloxidase − an enzyme involved in the immune response − were measured in females. No significant effect of fenoxycarb exposure was observed on the lipid profile of both newborn individuals and females, while reproductive success was severely impaired in exposed females. Particularly, precopulatory behavior was significantly reduced and fertilized eggs were unviable. This study highlighted the deleterious effects of the insect growth regulator fenoxycarb on gammarid reproduction, which could have severe repercussions on population dynamics.

Effects of an insect growth regulator and a solvent on honeybee (Apis mellifera L.) brood development and queen viability.

Honeybee toxicology is complex because effects on individual bees are modulated by social interactions between colony members. In the present study, we applied high doses of the insect growth regulator fenoxycarb to honeybee colonies to elucidate a possible interplay of individually- and colony-mediated effects regarding honey bee toxicology. Additionally, possible effects of the solvent dimethyl sulfoxide (DMSO) were assessed. We conducted studies on egg hatching and brood development to assess brood care by nurse bees as well as queen viability. Egg hatching was determined by the eclosion rate of larvae from eggs originating from colonies (i) treated with sugar syrup only, (ii) treated with sugar syrup containing DMSO and (iii) treated with sugar syrup containing fenoxycarb (dissolved in DMSO). To evaluate brood development, combs with freshly laid eggs were reciprocally transferred between colonies, and development of brood was examined in the recipient hive. Brood reared inside DMSO- and fenoxycarb-treated colonies as well as brood from DMSO- and from fenoxycarb-exposed queens showed higher mortality than brood not exposed to the chemicals. No differences were found in egg hatching among the treatments, but there was a higher variability of eclosion rates after queens were exposed to fenoxycarb. We also observed queen loss and absconding of whole colonies. Based on our results we infer that fenoxycarb has queen- as well as nurse bee-mediated effects on brood quality and development which can lead to the queen's death. There also is an effect of DMSO on the nurse bees' performance that could disturb the colony's equilibrium, at least for a delimited timespan.

Dynamic regulation ofDrosophilanuclear receptor activity in vivo

Nuclear receptors are a large family of transcription factors that play major roles in development, metamorphosis, metabolism and disease. To determine how, where and when nuclear receptors are regulated by small chemical ligands and/or protein partners, we have used a 'ligand sensor' system to visualize spatial activity patterns for each of the 18 Drosophila nuclear receptors in live developing animals. Transgenic lines were established that express the ligand binding domain of each nuclear receptor fused to the DNA-binding domain of yeast GAL4. When combined with a GAL4-responsive reporter gene, the fusion proteins show tissue- and stage-specific patterns of activation. We show that these responses accurately reflect the presence of endogenous and exogenously added hormone, and that they can be modulated by nuclear receptor partner proteins. The amnioserosa, yolk, midgut and fat body, which play major roles in lipid storage, metabolism and developmental timing, were identified as frequent sites of nuclear receptor activity. We also see dynamic changes in activation that are indicative of sweeping changes in ligand and/or co-factor production. The screening of a small compound library using this system identified the angular psoralen angelicin and the insect growth regulator fenoxycarb as activators of the Ultraspiracle (USP) ligand-binding domain. These results demonstrate the utility of this system for the functional dissection of nuclear receptor pathways and for the development of new receptor agonists and antagonists that can be used to modulate metabolism and disease and to develop more effective means of insect control.

Genetic differences in the production of male neonates in Daphnia magna exposed to juvenile hormone analogs

We studied the susceptibility of three genetically different strains of the cyclical parthenogen Daphnia magna (Cladocera, Crustacea) in producing male neonates following exposure to juvenile hormone analogs. In experiment 1, NIES, Clone A, and Belgium A strains were exposed to the insect growth regulators (IGRs) fenoxycarb or epofenonane in a 21-day reproduction experiment. Fenoxycarb exposure decreased the total number of neonates and increased production of male neonates in a concentration-dependent manner in the NIES strain. The decrease in the total number of neonates was so great in Clone A following fenoxycarb exposure that male neonates were not observed, even at the highest concentration, where the total number of neonates was only 2% of the control. In the Belgium A strain, male neonates were observed at a rate of about 20% following exposure to the highest fenoxycarb concentration, but the total number observed was small. Epofenonane did not decrease reproduction in the NIES and Belgium A strains as dramatically as did fenoxycarb, but the neonatal sex ratio changed in a concentration-dependent manner. Although the ratio of males was as low as about 10%, induction of male neonates was also observed in Clone A following epofenonane exposure. In experiment 2, gravid females were exposed to high concentrations (5 or 10 μg/l) of fenoxycarb or pyriproxyfen for 12 h. These treatments induced the production of male neonates in all strains, with a small decrease in the total number of neonates. Although induction of male neonates by juvenile hormones and their analogs was universal among genetically different strains, care is needed in interpreting the results of the 21-day reproduction tests, because decreased numbers of neonates at higher concentrations could obscure the presence of male neonates.

Optimization and validation of an enzyme immunoassay for the insect growth regulator fenoxycarb

A competitive enzyme-linked immunosorbent assay (ELISA) was developed for the quantitative detection of the insect growth regulator fenoxycarb. Polyclonal rabbit antisera, raised against protein conjugates of four haptenic derivatives of fenoxycarb, were utilized in immobilized antigen-based, competitive immunoassays. With ELISA systems that were both hapten- and carrier-heterologous, most antiserum titers fell in the range of 1:1000–1:30,000. Assay conditions, including concentrations of antisera and coating antigens, were optimized. The effect of pH, organic solvents, and various blocking agents was also investigated. A hapten-homologous and two hapten-heterologous indirect ELISAs allowed fenoxycarb determination in the range of 0.1–85 ng ml −1 with apparent IC 50 values of 1.2–2.8 ng ml −1. Cross-reactivities with a number of compounds (e.g. pesticides of related structure, hapten synthesis intermediates, fenoxycarb metabolite, photodegradation products) were determined, and the assay proved highly selective for fenoxycarb. In particular, no significant interference was found with selected pyrethroid and juvenile hormone analog insecticides, phenoxyacetic acid herbicides, and photodegradation products of fenoxycarb. Using spiked water samples, assay performance was validated by SPME/GC-MS.

Evaluation of an enzyme immunoassay for the detection of the insect growth regulator fenoxycarb in environmental and biological samples.

A competitive enzyme-linked immunosorbent assay (ELISA) for fenoxycarb was adapted for quantitative detection of this insect growth regulator in various environmental, agricultural, food and biological matrices. Environmental samples were taken from soil and surface waters in Hungary. The ELISA enabled fenoxycarb detection in surface waters in the 1.1-125 ng ml(-1) concentration range without sample cleanup. In contrast, soil produced a strong matrix effect due to humic acids and other soil components. Several fruit homogenates and commercial fruit juices (eg apple, pear, grape) were analyzed by the ELISA. The assay was found to be suitable for analysis of fenoxycarb in fruit juices diluted 1:40. Biological samples included insect, fish and bovine tissues. The ELISA was applied to detect fenoxycarb in various biological matrices from larvae of the silkworm, Bombyx mori L. The assay proved useful for the analysis of haemolymph diluted 1:10 or at higher dilutions. Fat body and whole body homogenates, however, caused severe matrix effects. Fenoxycarb was detected in liver homogenates (diluted 1:40) from fish treated with various doses of fenoxycarb, and the concentrations determined correlated with the applied doses. The method was used to analyze spiked bovine urine samples diluted 1:10 or at greater dilutions. Fenoxycarb content determined by the ELISA in water and fruit juice samples was validated using GC-MS with solid-phase microextraction (SPME) sample preparation. The results of these studies demonstrated both the value and limitations of the assay when used for monitoring fenoxycarb in environmental, food and biological samples.

COMPARATIVE COST OF CHEMICAL AND BIOLOGICAL WHITEFLY CONTROL IN POINSETTIA: IS THERE A GAP?

Abstract Cost is the principal constraint on the use of biological control against whiteflies in poinsettia (Euphorbia pulcherrima Willd. ex Koltz.) crops in the United States. Here we show that a new, lower release rate of the whitefly parasitoid Eretmocerus eremicus Rose and Zolnerowich (Hymenoptera: Aphelinidae), 0.5 females per plant per week, maintains whiteflies (Bemisia argentifolii Bellows and Perring) at harvest below the economic threshold of 2 live nymphs + pupae per leaf, when used in combination with two mid-crop applications of the insect growth regulator fenoxycarb (Precision®). Cost of this program (for 16.5 cm dia single stem pots, with 30,000 plants under protection) varies from 21 to 34 cents per plant for the season, for cropping periods from 12 to 18 weeks. Shipping costs are calculated and included in estimated costs. These values compare favorably to the real cost of whitefly chemical control incurred by Massachusetts poinsettia growers in fall of 2000, which was 14 cents for a 16.5...

Synthesis of haptens and protein conjugates for the development of immunoassays for the insect growth regulator fenoxycarb.

Sensitive and selective enzyme-linked immunosorbent assays (ELISAs) in the immobilized antigen format were developed for fenoxycarb (1), an insect growth regulator (IGR). The parent molecule [ethyl 2-(4-phenoxyphenoxy)ethylcarbamate] was derivatized at several positions to obtain haptens (2-5) that were used to produce protein conjugates and rabbit polyclonal antisera. Amino derivatives of fenoxycarb at the terminal and internal rings (2 and 3, respectively) were linked to carrier proteins by azo coupling. Carboxyalkyl-spacer groups were attached to the ethyl group and the nitrogen atom of the target compound (1) to obtain haptens 4 and 5, respectively. Hapten-homologous ELISAs based on protein conjugates of compounds 2 and 4 determined fenoxycarb in the mid-ppb range (IC(50), 102 and 95 ppb, respectively). A more sensitive hapten-heterologous ELISA (IC(50), 17 ppb; detection limit 0.5 ppb) involved the antiserum raised against a conjugate of hapten 2 and the plate-coating antigen obtained from compound 3. These assays displayed no significant interferences with photodegradation products of fenoxycarb, the IGRs methoprene and pyriproxyfen, and a variety of pesticides including the pyrethroids fenvalerate and cypermethryn, the phenoxyacetic acid herbicide 2,4-D, DDT, and the nitrodiphenyl ether herbicides acifluorfen and fluorodifen.

Sulphur-containing derivatives structurally related to fenoxycarb are potent growth inhibitors against the intracellular form of Trypanosoma cruzi

Sulphur-containing derivatives structurally related to the insect growth regulator fenoxycarb were shown to be extremely active antiproliferative agents against the amastigote form of Trypanosoma cruzi in in vitro assays. All of these drugs had previously been proved to be remarkably potent growth inhibitors against the epimastigote form of the parasite.

Growth inhibitory effect of juvenile hormone analogues on epimastigotes of Trypanosoma cruzi

Several compounds, structurally related to the insect growth regulator Fenoxycarb, exhibited interesting inhibition action to control proliferation of Trypanosoma cruzi, the parasite responsible for Chagas' disease. Some of these drugs were shown to be potent growth inhibitors of this parasite. All of these drugs had previously presented juvenoid activity on several non-related bug species such as Tenebrio molitor, Galleria mellonella, Dysdercus cingulatos, and Pyrrhocoris apterus.

Effects of Conventional Insecticides and Insect Growth Regulators on Fecundity and Other Life-Table Parameters of Micromus tasmaniae (Neuroptera: Hemerobiidae)

Effects of 3 conventional insecticides (methyl parathion, azinphos-methyl, cyper­ methrin) and 3 insect growth regulators (fenoxycarb, diflubenzuron, and tebufenozide) on life-table parameters of Micromus tasmaniae Walker were determined in adults derived from insecticide-treated larvae. The following parameters were compared with the control: sex ratio, longevity, sterility, and fecundity. Power analysis was used to increase the efficiency and the predictability of the life-table test. Diflubenzuron resulted in a higher proportion female lacewings. Longevity was reduced for females emerging from fenoxycarb- and diflubenzuron­ treated larvae. Total number of eggs was reduced for diflubenzuron- and fenoxycarb-treated lacewings, as weIl as the following generation oftebufenozide-exposed lacewings. Daily number of eggs was reduced for the diflubenzuron treatment. Peak egg production was increased for lacewings exposed to azinphos-methyl and was decreased for the following generation of tebufenozide-exposed lacewings. Diflubenzuron treatment resulted in an extended preovipo­ sition period. Oviposition periods were reduced for lacewings treated with fenoxycarb, di­ flubenzuron or azinphos-methyl as weIl as for the following generation of the tebufenozide treatment. The time to peak egg production was similar for all treatments. Methyl parathion, cypermethrin, and tebufenozide treatments showed no differences in any ofthe tested life-table parameters in the 1st generation. In summary, the insect growth regulators fenoxycarb and diflubenzuron had a more severe impact on life-table parameters than the 2 organophosphates and the pyrethroid. In future research, increased attention should be paid to long-term (e.g., the following generation) effects on life-table parameters.

Design, Synthesis, and Biological Evaluation of New Growth Inhibitors of Trypanosoma cruzi (Epimastigotes)

As a continuation of our project aimed at the search for new chemotherapeutic agents against Chagas' disease, several drugs structurally related to the insect growth regulator Fenoxycarb and the naturally occurring juvenile hormone of insects were designed, synthesized, and evaluated as antiproliferative agents against the parasite responsible of this disease. Isoprenoid derivatives (compounds 33, 34, 36, and 37) were potent growth inhibitors of Trypanosoma cruzi epimastigotes. In addition, taking into account the high activity observed for compound 30 and the inhibitory action of related compounds, the allyl ether moiety bonded at the polar extreme of these inhibitors proved to be a promising group for the design of new drugs.

Lacewings (Neuroptera: Hemerobiidae and Chrysopidae) and Integrated Pest Management: Enzyme Activity as Biomarker of Sublethal Insecticide Exposure

Specific activities of head acetylcholinesterase (AChE) and whole body glutathione-S-transferase (CST) were assayed as biomarkers of sublethal exposure to insecticides in larvae of 2 lacewing species, the Chrysopid Chrysoperla carnea Stephens and the Hemerobiid Micromus tasmaniae Walker. When M. tasmaniae was exposed to the organophosphates methyl-parathion or azinphos-methyl, the rate of AChE-inhibition was toxin-specific, exponentially dose-dependent, and increased within 24 h of exposure. Activity of AChE was less inhibited in C. carnea larvae, which reflected its higher tolerance to organophosphates in mortality tests. No inhibition of AChE activity (e.g., resulting from nonspecific binding to the enzyme) was observed following treatment with the pyrethroid cypermethrin and the insect-growth regulators fenoxycarb, diflubenzuron, and tebufenozide. The activity of GST increased significantly in M. tasmaniae larvae treated with sublethal doses of cypermethrin and decreased significantly in larvae treated with fenoxycarb. In contrast, no changes in GST activity were observed in C. carnea larvae for any of the compounds tested. Inhibition of AChE in lacewings proved a useful tool to study the impact of different organophosphates used in integrated pest management. However, further investigations are necessary to evaluate the potential applicability of GST activity as a biomarker in lacewings as different results are likely for different lacewing species, varying exposure times, repeated dose levels, different-aged larvae, and different substrates for the enzyme reaction.

Effects of fenoxycarb on ovarian development, spring fecundity and longevity in winterform pear psylla

AbstractThe insect growth regulator fenoxycarb prompts ovarian development in diapausing winterform pear psylla, Cacopsylla pyricola (Förster) (Homoptera: Psyllidae). We applied fenoxycarb to caged psylla in September, November, and December to test whether premature ovarian development reduced overwintering survival, spring fecundity, or spring longevity. Fenoxycarb prompted ovarian development in all treated psylla, with the largest effects occurring in the September‐treated insects. Recovery of live psylla in spring was 46–95% in treated insects and 72–92% in controls; overwintered insects from the fenoxycarb treatments survived field temperatures below −20°C despite having had mature ovaries. Fecundity and longevity of psylla were the same in treated and untreated insects, indicating that overwintering with mature ovaries did not cause reduced spring egglaying capacity. Several treated insects each deposited over 1900 eggs and survived more than 120 days. Due to their more advanced development, treated insects had higher oviposition rates than controls during the first 5 days after removal from the field. The largest impact on spring fitness was due to the effects of fenoxycarb on egg hatch. Eggs deposited on clean foliage by September‐treated females were less likely to hatch than eggs deposited by controls, suggesting that fenoxycarb affected developing eggs within the female. Prospects for using fenoxycarb in fall to control pear psylla appear to be limited.

Fenoxycarb-Induced Caste Differentiation and Mortality in Coptotermes formosanus (Isoptera: Rhinotermitidae)

In no-choice feeding bioassays, Formosan subterranean termites, Coptotermes formosanus Shiraki, exposed to the insect growth regulator (IGR) fenoxycarb underwent dosage- and time-dependent caste differentiation and mortality. Survival curves for 2 colonies indicated a decline in the number of workers over time at fenoxycarb concentrations ranging from 10 to 3,162 ppm, with a steeper decline generally at higher concentrations and after 2 wk exposure. Termite workers in the 2 colonies differed significantly in their response to the IGR. The worker population of colony A was reduced because of differentiation into intercastes, presoldiers, and soldiers. Reduction in the worker population of colony B resulted primarily from death of undifferentiated individuals. Termites accepted the treated blocks as a food source as evidenced by consumption data. Protozoan populations were reduced significantly in both colonies, but generally not eliminated in workers exposed to fenoxycarb for 4 wk. This study provides further evidence that responses of different termite colonies to IGRs can be extremely variable.

Reduction of Egg Hatch in Pear Psylla (Homoptera: Psyllidae) After Contact by Adults with Insect Growth Regulators

Pear psylla, Cacopsylla pyricola Foerster, adults that had contacted substrates treated with the insect growth regulators fenoxycarb or pyriproxyfen were allowed to oviposit on untreated plants. Eggs from treated insects had a reduction in hatch for up to 2 wk after treatment. The reduction was dependent on duration of contact with the treated surface. A pyriproxyfen-impregnated yellow tape has been developed and may assist in an attract-and-sterilize approach to management of pear psylla. Contact durations of as little as 5 s with this product reduced egg hatch in psylla for 1 wk after exposure.

Potential of Juvenoid Insect Growth Regulators for Managing Cotton Aphids (Homoptera: Aphididae)

Field and laboratory experiments demonstrated that the juvenoid insect growth regulators kinoprene and fenoxycarb and the insecticide synergist piperunyl butoxide- (PBO) slowed population growth, delayed adult emergence, and significantly alte-red cotton aphid. Aphis gossypii Clover. population structures. Normal cotton aphid populations have a large proportion of first-instar nymphs, fewer larger nymphs, and still fewer adults. Both juvenoid insect growth regulators and PBO decreased proportions of small nymphs and adults, resulting in populations concentrated at the larger nymphal age classes. In contrast, treatment With a conventional insecticide, dicrotophos, reduced numbers of all cotton aphid age classes similarly. Natural enemy populations were similar for control, juvenoid insect growth regulators, and PBO treatments, but were reduced by dicrotophos. The similarity in effects of juvenoid insect growth regulators and PBO suggests that PBO has insect juvenoid growth regulator activity. The ability of the juvenoid insect growth regulators and PBO to slow population growth and change population age distributions suggests their use in strategies for cotton aphid management that are not based strictly on mortality. Use of population-modulating chemicals for control of cotton aphid may be more efficient than current methods that kill the insects directly.

Control of the Codling Moth with Experimental Insecticides, 1994

Abstract The trial was conducted in a 30 yr old Bartlett pear block with chemicals applied to 4 single tree replicates arranged in a RCB. Materials were applied with conventional, high pressure hand gun equipment and sprayed to runoff or about 400 gpa. Each chemical was applied three times in a season-long, phenologically timed CM program. Two different timings were employed, based on CM biofix as determined by the first male moth captured in pheromone baited traps. A standard larvicidal timing was used in treatments receiving Guthion, A 303,603 and one treatment of RH 5992. This standard program consisted of a first application at about 250 degree days (dd) following biofix; the second 28 days later and the final at about 1250 dd, or 250 dd after the predicted emergence of second generation moths. Corresponding dates to the above schedule were 12—13 May, 9 Jun and 13 Jul. An ovicidal timing was employed for the insect growth regulator (IGR) fenoxycarb. The timings for fenoxycarb was a first application at 50-100 dd after biofix (27 Apr), the second 28 days after the first, and the third at 1050 dd (7 Jul). Evaluation of CM damage was made by recording the number of larval entries per 50 fruit per replicate at the end of the first CM generations (7-9 Jul) and again at harvest (17 Aug). Densities of PP immatures, spider mites (TSM, ERM) and PRM were estimated at three post-treatment intervals (20 Jun, 25 Jul, 16 Aug) by recording their numbers from brushing of 20 leaves (10 old, 10 new) per replicate. Adult PP and predators (lacewings, earwigs, and predaceous bugs) were estimated from two beating tray samples per replicate taken on 22 Jun, 27 Jul, and 19 Aug.