An improved method for determining the susceptibility ofRhizoglyphus robini andR. setosus (Acarina:Acaridae) to pesticides

Most bioassays used for determining the susceptibility ofRhizoglyphus mites face the problem of mite escapes, which leads to variation within replicates of treatments. An improved method uses surface tension of water to deter the bulb mites from escaping, and simplifies the food supply during the test with a piece of filter paper only. This method is described in detail and the susceptibility ofR. robini andR. setosus to several compounds measured by this method is demonstrated.     

Influence of temperature on cyhexatin resistance inTetranychus urticae (Acari: Tetranychidae)

The effect of post-treatment temperature on the toxicity of cyhexatin to cyhexatin-susceptible (S) and resistant (R) strains ofTetranychus urticae Koch was investigated. Females and developmental stages of both strains showed a positive temperature-toxicity coefficient with wettable powder (WP) and emulsifiable concentrate (EC) formulations of cyhexatin. Females of the S strain were 3.0× and 2.9× less susceptible at 15°C than at 28°C with the EC and WP formulations respectively, while females of the R strain were 5.2× and 23.6× less susceptible at 15°C with these formulations.     

Toxic effects of 12 pesticides on green lacewing,Chrysoperla nipponensis (Okamoto) (Neuroptera: Chrysopidae)

In this study, we formulated pesticides from 12 products, namely machine oil EC, imidacloprid WP, thiamethoxam WP, acetamiprid WP, methidathion EC, acequinocyl WP, clothianidin WP, deltamethrin EC, mancozeb WP, benomyl WP, difenoconazole WP, and bitertanol WP. These 12 pesticides were selected to determine their toxicity to green lacewing, Chrysoperla nipponensis, at the maximum field recommended dosage under laboratory conditions. Machine oil EC had extremely detrimental effects on eggs of C. nipponensis, resulting in 99% mortality (categorized as Class IV) when eggs were treated with machine oil EC by dipping. Mean larval corrected mortalities (%) for methidathion EC and deltamethrin EC were 62.5% (categorized as Class II) and 87.5% (categorized as Class III) respectively, when larvae were topically treated. As a result of dipping treatments of pupae with pesticides, machine oil EC and thiamethoxam WP were classified as slightly harmful (categorized as Class II). Methidathion EC showed high toxicity, resulting in a total effect index rate 100% (categorized as Class IV). Taxonomical notes of the genus Chrysoperla and C. nipponensis are reviewed here.     

Resistance levels ofTyphlodromus occidentalis (Acari: Phytoseiidae) from Washington apple orchards to ten pesticides

Nine Washington populations ofTyphlodromus (=Metaseiulus) occidentalis Nesbitt collected from commercial apple orchards were surveyed for resistance to ten pesticides. A susceptible population collected from wild blackberry which had no history of pesticide exposure was used in estimating resistance development. All populations from apple orchards were tolerant to high concentrations of azinphosmethyl, diazinon, endosulfan and propargite. Resistance was also apparent to these materials, especially for azinphosmethyl which produced over 100-fold resistance levels for several populations. Moderate to low levels of tolerance and resistance development were seen to field concentrations of cyhexatin, formetanate and carbaryl. Fenvalerate, methomyl and methidathion were all very toxic at field concentrations and little evidence of resistance development was apparent. The use of fenvalerate, methomyl or methidathion at a time whenT. occidentalis is active would likely abrogate favorable biological control of pestiferous orchard mites.Scientific Paper No. 7502     

Enzyme polymorphisms in Rhizoglyphus robini and R. echinopus and their application in paternity analysis

Cellulose acetate electrophoresis (CAE) was used to investigate enzyme polymorphism in two congeneric species of Acaridae, Rhizoglyphus robini and R. echinopus. Using homogenates of individual mites, 27 enzymes were examined in two buffers. Five enzymes showed interspecific polymorphism, 15 exhibited intraspecific variation inR.robini and 12 in R.echinopus. Polymorphic PGDH was used to analyse paternity in progeny of R.robini females that had mated with two males. The second male sired on average 67% of progeny. Enzyme polymorphism can be used to study poorly known aspects of acarid mites biology, such as their mating systems and population structure under natural conditions and in stored foods. This revised version was published online in August 2006 with corrections to the Cover Date.     

Toxicity of organophosphorus insecticides to diazinon-reversely-selected,pesticide-pressure-relaxed and field-resistant strains of bulb miteRhizoglyphus robini (Acari: Acaridae)

Some field strains ofRhizoglyphus robini in Taiwan, developed high multi-resistance to organophosphorus insecticides including diazinon, dimethoate, fenitrothion, malathion, methyl parathion, omethoate, parathion and prothiophos. The organophosphorus resistance of bulb mites remained stable when the pesticide pressure was relaxed. Changes of the susceptibility to several organophosphorus insecticides for a strain after relaxation of selection pressure for 3 and 27 generations were compared. Since susceptible bulb mites could not be found in the field, a reverse selection for susceptibility to diazinon was designed and conducted. Although heterogeneity was observed in the progeny of the mixed populations, the susceptibility of these reversely-selected mites increased slowly. A stable and susceptible strain was obtained after 26 generations of selection.Contribution No. 1412 from Taiwan Agricultural Research Institute.     

Amplification of single bulb mites by nested PCR: Species-specific primers to detect Rhizoglyphus robini and R. setosus (Acari: Acaridae)

A major problem of using molecular amplicons for DNA amplification in mite systematics is that sufficient template cannot always be acquired from an individual mite. To solve this problem, we developed a nested PCR for DNA amplification of single Rhizoglyphus robini and R. setosus bulb mites. A dilution up to 10⁵ of the DNA from a single egg, larva, nymph or adult contained enough templates for amplification of the target ribosomal region. However, the use of specific primers in the second PCR is necessary to reduce the generation of non-target DNAs from symbiotic organisms. Identification of bulb mites collected from seven sampling locations in Taiwan or of bulb mites that were used in simulated experiments in the presence of host plant tissues was unambiguous with specific PCR primers.     

Monooxygenase Activity in Methidathion Resistant and Susceptible Populations of Amblyseius womersleyi (Acari: Phytoseiidae)

The purpose of this study was to evaluate the cytochrome P450-dependent monooxygenase activities in methidathion resistant and susceptible strains of Amblyseius womersleyi Schicha. Artificial laboratory selections for resistance and susceptibility to methidathion were performed in an organophosphate resistant strain of A. womersleyi (Kanaya strain). Selections for susceptibility were also performed in a susceptible strain of this predaceous mite (Ishigaki Strain). After the selection process, the LC₅₀ of methidathion for the selected strains of A. womersleyi were 816 mg/l (Kanaya R), 4.61 mg/l (Kanaya S) and 1.59 mg/l (Ishigaki S). The monooxygenase activities were determined biochemically by the O-deethylation of 7-ethoxycoumarin (7-EC). The monooxygenase activity in adult females of Kanaya R strain (51.1 pmol/30 min/mg protein) was 3.60- and 5.42-fold higher than the activity observed for Kanaya S and Ishigaki S strains, respectively. Significant correlation between monooxygenase activity and LC₅₀ (mg/l) of methidathion was observed analyzing 16 populations of A. womersleyi with different susceptibilities to methidathion. Monooxygenase activity was also evaluated in different life stages (egg, larva, protonymph, deutonymph and adult) of A. womersleyi. The lowest activity was observed for the larval stage, which presented the highest susceptibility to methidathion. Protonymph, deutonymph and adult presented the highest monooxygenase activities. These stages were the most tolerant to methidathion. Monooxygenase activities of the Kanaya R strain were higher than of the Kanaya S strain in all developmental stages. The present study can be helpful for the implementation of a program involving release of insecticide-resistant populations of A. womersleyi in the field. The monooxygenase activity determination is easier and quicker than the estimation of LC₅₀, requiring fewer mites.     

Efficacy of various synthetic pyrethroid-impregnated encasement materials against house dust mite under laboratory conditions

The acaricidal activity of synthetic pyrethroid and benzyl benzoate against Dermatophagoides pteronyssinus was examined in the laboratory, using a specially designed test set up. On the basis of median lethal dose (LD₅₀) values, the compound found to be most toxic to D. pteronyssinus was benzyl benzoate (LD₅₀ = 50 mg/m²), followed by permethrin (LD₅₀ = 76.7 mg/m²), deltamethrin (LD₅₀ = 146.7 mg/m²), esbioallenthrin (LD₅₀ = 186.6 mg/m²) and lamdacyhalothrin (LD₅₀ = 756.6 mg/m²). Very low toxicity was observed with bifenthrin (LD₅₀ = 5157.8 mg/m²). A laboratory control trial was also carried out to compare the acaricidal activity (residual effect) of four pyrethroids impregnated on woven and non-woven encasement materials against house dust mites during a 4-month period. Of the pyrethroids used in this study, esbioallenthrin demonstrated the highest acaricidal activity, and of the pyrethroid impregnated materials, the non-woven encasement material was more effective than the woven encasement material.     

Susceptibility ofEuseius addoensis addoensis (Acari: Phytoseiidae) to field-weathered residues of insecticides used on citrus

Comparisons of the toxicities of field-weathered residues of insecticides to adult femaleEuseius addoensis addoensis (McMurtry) were conducted by bioassay using leaf disks. The products tested are registered for the control of certain insect pests on citrus in South Africa. The treatments which caused the highest mortality, in order of decreasing toxicity, were: methiocarb plus sugar, cypermethrin plus profenofos, fenpropathrin plus phenthoate, methomyl, methidathion, phenthoate, prothiofos plus oil, and profenofos. Most of these treatments also had prolonged residual effects. Endosulfan, which caused only intermediate mortality to adult female mites, had a very persistent residue. Sublethal effects of insecticide residues on mite fecundity were negligible in most cases.     

Pesticide-induced mite outbreaks: pyrethroids and spider mites

Factors inducing outbreaks of spider mites (Acari: Tetranychidae) following use of pyrethroid insecticides are reviewed. Differentials in direct toxicity between spider mites and phytoseiid (Acari: Phytoseiidae) predators provide one explanation. Wide variation exists between pyrethroids in their direct toxicity toTetranychus urticae. The acaricidal action of pyrethroids is largely controlled by the amount of irritancy or repellency induced by the respective chemicals. Laboratory assays for repellency reflect field results. Repellent activity induces spider-mite dispersal to either recolonise plants free of residues or leave the treated habitat. Pyrethroids may also affect reproduction and development rates. Residual activity is likely to vary, so limiting any general theory of spider-mite responses to pyrethroids. Variable responses to pyrethroids, strain variation and development of behavioural resistance also add complications to understanding outbreaks. Future research needs are identified.     

Molecular identification and phylogenetic analysis of economically important acaroid mites (Acari: Astigmata: Acaroidea) in Korea

Molecular diagnosis is highly valuable for the species identification of microscopic mites. Here, we collected some economically important mites of the superfamily Acaroidea from various stored products in Korea. Those nucleotide sequences of ribosomal second internal transcribed spacer (ITS2) and the mitochondrial cytochrome oxidase subunit I (COI) regions were determined by PCR using universial primers. In nucleotide sequence comparison at GenBank database, seven species including Rhizoglyphus robini, R. echinopus, Sancassania sp. Acarus siro, Tyrophagus putrescentiae, T. similis in Acaridae and Suidasia medanensis in Suidasiidae were identified. Particularly, COI sequences of R. robini, R. echinopus, Sancassania sp. and T. similis were firstly determined. Our results suggest that the phylogenetic relationships inferred from the ITS2 region, rather than COI region, were similar to those derived based on their morphological classification. Our study provides molecular information for the identification and phylogenetic relationship of acaroid mites in Korea.     

Physiological resistance and behavioral avoidance responses to residues of four pesticides by six spider mite populations

Six twospotted spider mite populations were assayed for their levels of physiological resistance and behavioral avoidance to residues of four synthetic pesticides. Mortality could not be estimated for bifenthrin and fenvalerate (synthetic pyrethroids) as mites effectively avoided treated surfaces, however significant between-population differences in mortality were detected for chlordimeform and cyhexatin. Considerable variation in walkoff and spindown behavioral response to sub-lethal doses of pesticides was observed among populations within compounds, and within populations among compounds. Within-compound walkoff and spindown behavioral response varied among all mite populations. Few significant between-compound correlations were significant, indicating that spider mites responded differently to the four pesticides. The hypothesis that physiological resistance is negatively correlated with behavioral avoidance was tested. Of the four possible negative correlations between physiological resistance and behavioral avoidance for chlordimeform and cyhexatin, only the correlation between cyhexatin-induced mortality and spindown response was significant. Comparisons of physiological resistance and behavioral avoidance of chlordimeform and cyhexatin by specific pairs of populations did not consistently find these two characters to be related. In a related experiment, the magnitude and direction of the correlation between physiological resistance and behavioral avoidance following selection for increased physiological tolerance to cyhexatin was compared in a highly resistant and a susceptible population of the twospotted spider mite. Mortality in the susceptible population at 2 ppm cyhexatin was similar to mortality in the resistant population at 250 ppm after 72h exposure (ca. 12%). However, at these concentrations, the resistant population exhibited much higher avoidance of the compound through walkoff response.     

Resistance of Brevipalpus phoenicis (Geijskes) (Acari: Tenuipalpidae) to acaricides that inhibit cellular respiration in citrus: cross-resistance and fitness cost

Acaricides that inhibit cellular respiration play an important role in the control of Brevipalpus phoenicis (Geijskes) in citrus groves in Brazil. Studies were conducted to evaluate: (a) the variability in the susceptibility among B. phoenicis populations collected from citrus groves to cyhexatin, azocyclotin, propargite and sulphur; (b) cross-resistance relationships between propargite and azocyclotin, cyhexatin, dinocap, pyridaben and sulphur; and (c) the fitness cost associated with propargite resistance in B. phoenicis under laboratory conditions. A residual-type contact bioassay was used to characterize the susceptibility. The susceptibility was estimated with diagnostic concentrations based LC(95) of each acaricide. The cross-resistance was evaluated by characterizing the concentration-mortality responses of susceptible (S) and propargite-resistant (Propargite-R) strains. The fitness cost was evaluated by measuring the biological parameters of S and Propargite-R strains on citrus fruits at 25 +/- 1 degrees C and fotophase of 14h. Significant differences in the susceptibility of B. phoenicis were detected at diagnostic concentration of cyhexatin (survivorship from 16.3% to 80.5%), azocyclotin (from 3.0% to 15.0%), propargite (from 1.0% a 71.6%) and sulphur (from 9.0% to 82.6%). A low intensity of cross-resistance was detected between propargite and the acaricides azocyclotin (1.8-fold), cyhexatin (4.6-fold), dinocap (3.5-fold) and pyridaben (3.5-fold). On the other hand, the intensity of cross-resistance to sulphur (> 111-fold) was very high. There was no fitness cost associated with B. phoenicis resistance to propargite, based on biological parameters evaluated. Therefore, the use of these acaricides should also be done very carefully in resistance management of B. phoenicis to acaricides.     

EFFECTS OF AGROCHEMICALS ON BROWN PL THOPPER NILAPARVATA LUGENS (STÅL) AND ITS PREDATORY ENEMIES*

Abstract The effects of agrochemicals on brown planthopper (BPH), Nilaparvata lugens, three predators were studied. The results showed that with recommendatory concentrations of 0. 30preti-lachlor EC could cause high mortalities to adults of BPH, Microvelia horvathi and Cyrtorhinus lividipennis; 0. 50 butachlor EC had significant lethal effects on adults of M. horvathi and C. lividipennis, but had negligible effects on BPH and Erogonidium graminicolum; and 40% acetochlor WP had no obvious killing effect to other insects mentioned above, except C. lividipennis. Effects of 0. 50 quincolorac WP, 0. 10 bensulfuron methyl WP, 0. 10 cyclosulfumuron WP and 0. 20 daofuxing WP on these insects were insignificant. As far as fungicides were concerned, 2nd-3rd instar nymphs of BPH and adults of Microvelia horvathi, and Cyrtorhinus lividipennis were obviously sensitive to 0. 40 IBP EC, but 0. 75 chlorothalonil WP 0. 20 tricyclazol WP, 0. 50 carbendazin WP, 0. 25 yekuling WP, 0. 5 jinggangmycin SL and 0. 20 yeqing shuang WP had no evident effects on them. 0. 4635 urea G had also no obvious insecticidal activity on adults of these insects. The results of toxic regression expression and LC₅₀ of IBP, pretilachlor, butachlor and acetochlor to BPH and its predators were determined.     

Laboratory evaluation of some plant essences to control Varroa destructor (Acari: Varroidae)

This research was conducted to evaluate acaricidal effects of some plant essences on Varroa mites and the possibility of their usage for Varroa control. First, live Varroa mites were obtained from adult honeybees with CO₂ in a newly designed apparatus. Thyme, savory, rosemary, marjoram, dillsun and lavender essences at concentrations of 2 and 1g/100g (w/w), caused a mite mortality rate of more than 97% and 95%, respectively. Also spearmint at 2g/100g was able to kill more than 97% of Varroa mites. When sprayed on worker honeybees infected with mites, thyme, savory, spearmint and dillsun essences at 2g/100g (w/w) caused 43–58% Varroa mortality. Toxicity of thyme, savory and spearmint essences for worker honeybees was not significantly different from that of controls (acetone and water), but dillsun essence caused 12% honeybee mortality. These results showed that essences of thyme, savory and spearmint have acaricidal properties that could be used for controlling Varroa in honeybee colonies. This revised version was published online in July 2006 with corrections to the Cover Date.     

Candidate natural enemies for control of Rhizoglyphus robini Claparède (Acari: Astigmata) in lily bulbs: exploration in the field and pre-selection in the laboratory

To find suitable candidates for biological control of the bulb mite, Rhizoglyphus robini Claparède (Acari: Astigmata) on lilies, exploration was undertaken in areas where the bulb mite is an established pest (The Netherlands, Taiwan and Japan). Among the predators, found in association with R. robini in the field and under storage conditions, mesostigmatic mites predominate. The most abundant species were Hypoaspis aculeifer (Canestrini), Lasioseius bispinosus Evans and Parasitus fimetorum (Berlese). These predators appeared to feed and reproduce on a diet of exclusively R. robini and they were able to control the bulb mite in small-scale population experiments initiated with a 1:20 predator-prey ratio. Under laboratory conditions corresponding to lily bulb propagation (lily scales mixed with vermiculite and stored at 23°C and >90% RH) the laelapid mite, H. aculeifer, was the most effective predator; the ascid predator, L. bispinosus, was much less effective, but being relatively small and being successful in attacking the juvenile stages of the bulb mite it may be better able to search for bulb mites hidden inside the lily bulb. The parasitid predator, P. fimetorum, failed to control the bulb mite when vermiculite was used as a medium, but turned out to suppress this prey when peat was used instead. Various strains of H. aculeifer or closely related species were compared with respect to their impact and performance on bulb mites as prey: two Dutch strains, one obtained from Breezand and the other from 'tZand, a Taiwanese strain, a German strain that in contrast to the previously mentioned strains was not collected from lily bulbs, but from agricultural areas near Bremen and, in addition, a Canadian strain of a related species (Hypoaspis miles Berlese), known to control sciarid fly larvae. These comparative experiments showed that H. miles died out without noticeable impact on the bulb mite population whereas all strains of H. aculeifer were able to suppress the bulb mites to very low numbers. However, the numerical responses of the H. aculeifer strains differed in that those collected in association with the pest (Breezand > Taiwan > 'tZand) were superior to the strain from Bremen. These results do not provide support to the Hokkanen and Pimentel hypothesis, which states that predators forming an evolutionary new association with the pest are often more effective in biological control.     

Inhibition of ATP dephosphorylation by acaricides with emphasis on the anti-ATPase activity of the carbodiimide metabolite of diafenthiuron.

3-(2,6-Diisopropyl-4-phenoxyphenyl)-1-tert-butylcarbodiimide (DFCD), a toxic metabolite and photodegradation product of the propesticide diafenthiuron, and dicyclohexylcarbodiimide (DCCD), a commonly used biochemical inhibitory probe, inhibited Mg(2+)-, Na+K(+)-, and Ca2+Mg(2+)-ATPase activities in preparations from bulb mites (Rhizoglyphus echinopus (Fumouze and Robin)), twospotted spider mites (Tetranychus urticae Koch), and bluegill (Lepomis macrochirus Rafinesque) brain. DCCD was more active than DFCD, but neither carbodiimide was very potent. A possible exception occurred with DCCD, which caused 100% inhibition of bulb mite oligomycin-sensitive Mg(2+)-ATPase activity at a concentration of 0.1 mM. Using house fly, Musca domestica L., thorax mitochondrial Mg(2+)-ATPase, we showed that the binding domain for both carbodiimides was in the F0 portion of the enzyme, probably the transmembrane proton channel which is the known site of DCCD binding in proton-translocating ATPases. Certain other specific acaricides were inhibitors (greater than 50% at 0.1 mM) of ATPase preparations from bulb mites. These acaricides included chloropropylate, bromopropylate, oxythioquinox, cyhexatin, and flubenzimine (Mg2+ and Na+K(+)-ATPase), and ovex, chlorbenside, and propargite (Mg(2+)-ATPase). The role of ATPase inhibition in the modes of acaricidal and insecticidal actions of diafenthiuron, the two carbodiimides, and the other compounds is discussed.     

Absence of non-target effects of two Bacillus thuringiensis coleopteran active δ-endotoxins on the bulb mite, Rhizoglypus robini (Claparède) (Acari, Acaridae)

Transgenic crops with plant‐incorporated protectants are often more specific than synthetic insecticides and have the potential to reduce impacts on non‐target organisms. In this study we assessed the impact of Cry3Aa and Cry3Bb1 coleopteran‐active δ‐endotoxins on the bulb mite, Rhizoglypus robini. The effect of Cry3Aa prototoxin in solutions of the biopesticide Novodor^® on mite survival was assessed in laboratory studies. Survival of R. robini exposed to Cry3Aa in a short‐term contact and ingestion experiment was not affected, although R. robini was significantly affected by the insecticide Fipronil^® used as a positive control. Similarly, R. robini exposed in a longer duration feeding trial to the Cry3Aa toxin in artificial diet were also not significantly affected. When Cry3Aa was tested on the positive control insect, Leptinotarsa decemlineata, reduced weight of larvae and increased mortality was recorded. The effect of Cry3Bb1 toxin in transgenic corn tissues on R. robini food choice was assessed in a laboratory study. In no‐choice tests a greater proportion of R. robini were found on garlic roots than on Cry3Bb1 transgenic corn and a near‐isogenic non‐transgenic corn. In a choice test, more R. robini was recovered on garlic roots than on either corn variety, and on Cry3Bb1 corn than on non‐transgenic corn. In large field plots using specific mite traps across the growing season, R. robini mite populations were not significantly different between Cry3Bb1 corn and non‐transgenic corn alone or non‐transgenic corn treated with different combinations of two insecticides. Our results, combined with results from other studies, suggest that transgenic plants expressing the Cry3Aa or Cry3Bb1 Bacillus thuringiensis toxins are likely to have negligible impact on R. robini mite populations.     

Conservation of natural enemy fauna in citrus canopies by horticultural mineral oil: Comparison with effects of carbaryl and methidathion treatments for control of armored scales

Abstract The disruptive effects of an nC24 horticultural mineral oil (HMO) and two other insecticides (carbaryl and methidathion) on two armored scale insects and natural enemy fauna were evaluated in two citrus orchards. In the first orchard, all three spray treatments and non-sprayed controls were distributed separately among individual trees in one block, whereas in the second orchard the control, HMO and methidathion treatments were applied at the level of whole blocks. The results in both orchards were similar. All three of the spray treatments had a lethal effect on the red scale insects and the effects of HMO and methidathion tested at the second site lasted for at least 19 weeks and for purple scale, at least 50 weeks. HMO did not have a significant effect on the abundance and species diversity of parasitoids and (for the most part) did not affect coccinellids and predatory mites. Both carbaryl and methidathion caused significant ongoing disruption to the above groups. No treatment resulted in the resurgence of the scale insects or the outbreak of phytophagous mites, even where whole blocks were sprayed.