Effects of neonicotinoid insecticides on the bionomics of twospotted spider mite (Acari: Tetranychidae)

Previous reports indicate that applications of imidacloprid, a neonicotinoid insecticide, can lead to population buildups of twospotted spider mite, Tetranychus urticae Koch, in the field. Moreover, laboratory studies showed enhanced fecundity of T. urticae after an imidacloprid treatment. In this study, experiments were conducted in the greenhouse to investigate the potential effects of imidacloprid and several other neonicotinoid insecticides on fecundity, egg viability, preimaginal survivorship, and sex ratio of T. urticae (German strain WI) on French beans, Phaseolus vulgaris L. Four insecticides, i.e., imidacloprid (Confidor 200SL), thiacloprid (Calypso 480 SC), acetamiprid (Mospilan 70 WP), and thiamethoxam (Actara 25 WG), were tested at field-relevant (100, 120, 125, and 95 ppm) and sublethal doses (10, 12, 12.5, and 9.5 ppm), respectively. Both spray and drench applications were tested. At field-relevant doses, fecundity of T. urticae decreased and was lower in the treatments compared with the untreated control, whereas preimaginal survivorship and proportion of female offspring (i.e., sex ratio) were lower compared with the control. At sublethal doses, no significant differences were found among the treatments. Data on egg viability, preimaginal survivorship, and sex ratio at sublethal doses followed the same trends as at field-relevant doses. In an additional experiment, the metabolism of imidacloprid into monohydroxy-imidacloprid, olefine, guanidine, and 6-chloronicotinic acid was compared with the oviposition pattern of T. urticae. These findings are discussed with regard to previous laboratory and field observations of imidacloprid-induced fertility increases in T. urticae.     

Approaches for sampling the twospotted spider mite (Acari: Tetranychidae) on clementines in Spain

Tetranychus urticae Koch (Acari: Tetranychidae) is an important pest of clementine mandarins, Citrus reticulata Blanco, in Spain. As a first step toward the development of an integrated crop management program for clementines, dispersion patterns of T. urticae females were determined for different types of leaves and fruit. The study was carried out between 2001 and 2003 in different commercial clementine orchards in the provinces of Castelló and Tarragona (northeastern Spain). We found that symptomatic leaves (those exhibiting typical chlorotic spots) harbored 57.1% of the total mite counts. Furthermore, these leaves were representative of mite dynamics on other leaf types. Therefore, symptomatic leaves were selected as a sampling unit. Dispersion patterns generated by Taylor's power law demonstrated the occurrence of aggregated patterns of spatial distribution (b > 1.21) on both leaves and fruit. Based on these results, the incidence (proportion of infested samples) and mean density relationship were developed. We found that optimal binomial sample sizes for estimating low populations of T. urticae on leaves (up to 0.2 female per leaf) were very large. Therefore, enumerative sampling would be more reliable within this range of T. urticae densities. However, binomial sampling was the only valid method for estimating mite density on fruit.     

Temperature-dependent fumigant activity of essential oils against twospotted spider mite (Acari: Tetranychidae)

Fumigant activity of 34 commercial essential oils was assessed on female adults and eggs of twospotted spider mite, Tetranychus urticae Koch (Acari: Tetranychidae) at three temperatures (5, 15, and 25 degrees C). Common thyme, cinnamon, and lemongrass oils were equally effective on twospotted spider mite adults showing 85.8-100% mortality at 5 and 10 microl/liter air at 25 degrees C. At a lower temperature of 15 degrees C, lemongrass and peppermint resulted in > or =90% mortality of adults at 10 microl/liter air. Only lemongrass was relatively active at 5 microl/liter air, at 15 degrees C. At 5 degrees C, lemongrass and peppermint caused significantly higher adult mortality than controls but only at 10 microl/liter air. Common thyme oil showed the highest ovicidal activity at 5 microl/liter air at 25 degrees C. Among the main components of common thyme and lemongrass oils, citral was lethal to twospotted spider mite adults at all tested temperatures. Carvacrol, thymol, and citral caused the same inhibitory effects on the hatch of twospotted spider mite eggs at 25 degrees C. However, citral was more active than other compounds to twospotted spider mite eggs at 15 degrees C. Therefore, we conclude that citral has the best potential for development as a fumigant against twospotted spider mite on agricultural products harvested late in the growing season.     

Impact of twospotted spider mite (Acari: Tetranychidae) on growth and productivity of glasshouse cucumbers

Experiments were conducted to assess the damage of the glasshouse cucumber by twospotted spider mite, Tetranychus urticae Koch, and to investigate when the economic yield begins to decrease after T. urticae infestations. To assess the damage, dry matter partitioning in the cucumber plant was quantified and plant growth analyses were conducted at four different T. urticae infestation levels. T. urticae infestations decreased leaf productivity by reducing the total number of leaves per plant. Approximately 14% reductions of total leaf areas could result in significant yield loss. The decreased leaf productivity by T. urticae feeding caused biomass reductions and altered the pattern of dry matter partitioning in the plant; damaged plants accumulated more dry matter in the leaf, and partitioning of dry matter to fruits was hindered. The economic yield of cucumber began to significantly decrease as early as 4 wk after heavy mite infestations. This study also showed the seasonal differences in T. urticae-cucumber damage interactions among mite infestation levels.     

Genetic transfer of a twospotted spider mite (Acari: Tetranychidae) repellent in tomato hybrids

Lycopersicon hirsutum Dunal is very resistant to arthropod herbivory, and research on causes of resistance has often implicated trichomes and their secretions. To better understand relationships among resistance, repellency, and 2,3-dihydrofarnesoic acid, a trichome-borne sesquiterpenoid spider mite repellent, two tomato, Lycopersicon esculentum Miller, varieties were interbred with a highly resistant, spider mite repellent accession (LA1363) of L. hirsutum. Backcross and F2 generations were produced with each tomato variety. Whole leaves of 99 hybrids were bioassayed with twospotted spider mites, Tetranychus urticae Koch, allowing selection of six hybrids (two susceptible and four resistant) for each generation of each family. When these 24 hybrids were characterized for spider mite repellency with thumbtack bioassays, two hybrids had repellent leaflets, demonstrating that repellency was genetically transferred to interspecific tomato hybrids. Leaflet washes containing trichome secretions from each of three hybrids, including the two having repellent leaflets, were repellent in bridge bioassays. For the two hybrids having repellent leaflets and leaflet washes, removal of trichome secretions by dipping leaflets in methanol eliminated leaflet repellency. 2,3-Dihydrofarnesoic acid was present in trichome secretions of the hybrids having leaflet repellency, and it also was present in secretions of other hybrids, indicating that its presence is essential, but not sufficient for leaflet repellency. With regard to resistance, 16 of the hybrids tested had been identified as resistant in a whole leaf bioassay, but only two had repellent leaflets, indicating that other mechanisms of resistance are present in the resistant L. hirsutum parent.     

Leaf cell and tissue damage of cucumber caused by twospotted spider mite (Acari: Tetranychidae)

To quantify the damage caused by twospotted spider mite, Tetranychus urticae Koch, feeding on cucumber, Cucumis sativus L., leaf-cell and -tissue damage was assessed. On the abaxial leaf surface, adult T. urticae could feed through the spongy parenchyma and part of the palisade parenchyma of the leaf, while immature T. urticae could feed only through the sponge parenchyma. T. urticae punctured individual epidermal cells and consumed the contents of the mesophyll cells. Injured leaves had more empty space in the spongy parenchyma and fewer chloroplasts per cell. Damage also occurred even in the adjacent uninjured parenchyma cells without additional T. urticae feeding injury. Net photosynthetic rate, total chlorophyll content, and greenness of the leaf were significantly affected by feeding as quantified by mite-days. The percent loss of these parameters increased linearly or nonlinearly as mite-days increased, regardless of mite developmental stages. At 1,000 mite-days per 6 cm2, net photosynthetic rate was reduced by approximately 50 and 95%, total chlorophyll content was reduced by approximately 55 and 80%, and greenness was reduced by approximately 50 and 80% by feeding by immature and adult T. urticae, respectively.     

Characterization of vitellin protein in the twospotted spider mite, Tetranychus urticae (Acari: Tetranychidae)

In mites, vitellogenin synthesis, regulation and uptake by the oocytes as vitellin remain practically unknown. Although a partial sequence of the gene is now available, no previous studies have been conducted that describe the native vitellin protein in mites. The objective of this study was to characterize vitellin in the twospotted spider mite, Tetranychus urticae. The native twospotted spider mite vitellin migrated as a single major band with a molecular weight of 476 ± 14.5 kDa as compared to 590 ± 25.5 kDa for vitellin from the American dog tick, Dermacentor variabilis. However, isoelectric focusing analysis of native spider mite vitellin showed five bands with pI values slightly acidic to neutral (pH 5.8, 6.2, 6.7, 7.0 and 7.2), as is the case for insect and tick vitellins. Reducing conditions (SDS-PAGE) also revealed multiple subunits ranging from 290.9 to 3.6 kDa and was similar to that found in D. variabilis. Spider mite vitellin weakly bound lipids and carbohydrates compared to the tick. Unlike D. variabilis, the spider mite egg yolk protein does not bind heme. The significance of non-heme binding in mites is discussed.     

Fecundity in twospotted spider mite (Acari: Tetranychidae) is increased by direct and systemic exposure to imidacloprid

The effect of imidacloprid on fecundity in twospotted spider mites, Tetranychus urticae Koch, was investigated in laboratory experiments using individual females on bean leaf discs. Mites were directly exposed to spray formulations of imidacloprid or fed on discs cut from a systemically treated bean plant. Imidacloprid-treated T. urticae produced 10-26% more eggs during the first 12 d of adult life and 19-23% more during adulthood compared with a water-only treatment. Increased egg production occurred immediately after exposure and lasted for about 15 d in sprayed mites. In mites exposed to imidacloprid by ingestion, increased egg production was not apparent until after 6 d and lasted until about day 18. Longevity was significantly greater in mites that ingested imidacloprid but not in sprayed mites. The significance and importance of imidacloprid-stimulation of fecundity in T. urticae to pest management in crop systems like hops, which routinely use this insecticide, is discussed.     

Effects of soil moisture and temperature on reproduction and development of twospotted spider mite (Acari: Tetranychidae) in strawberries

The effects of soil moisture and temperature on the reproduction of twospotted spider mite, Tetranychus urticae Koch (Acari: Tetranychidae), were examined in laboratory and field tests in strawberries, Fragaria x ananassa Duchesne, in Florida. Different soil moisture levels (low, moderate, and high) were compared to determine how soil moisture affects the reproduction and development of twospotted spider mite. In addition to soil moisture, different irrigation techniques (drip versus drip/overhead) were compared to determine their effects on twospotted spider mite reproduction as well as the incidence of angular leaf spot, Xanthomonas fragaria Kennedy & King disease. Similar studies were conducted to determine how different temperatures (18, 27, and 35 degrees C) affect the reproduction and development of twospotted spider mites. In the laboratory, low soil moisture as well as temperatures >27 degrees C promoted twospotted spider mite development. A similar trend was observed in a field study with low soil moisture promoting twospotted spider mite reproduction during the early season (11 November--8 December). Irrespective of moisture levels, a significantly higher incidence of X. fragaria was recorded in treatments with drip/overhead irrigation systems compared with drip irrigation. Implications for management of soil moisture levels are discussed with respect to the abundance of twospotted spider mite and X. fragaria in strawberries.     

Host plant-induced changes in detoxification enzymes and susceptibility to pesticides in the twospotted spider mite (Acari: Tetranychidae)

Adult female twospotted spider mites, Tetranychus urticae Koch, reared on lima bean plants were moved to cucumber, maize, or new lima bean plants (the latter being a control) and evaluated after 24 h or 7 d for changes in susceptibility to three pesticides and in levels of related detoxification enzymes. The largest and most consistent changes were observed in mites feeding on cucumber. Susceptibility of mites on cucumber to the synthetic pyrethroids bifenthrin and lambda-cyhalothrin was greater than that of mites reared on lima bean and maize after only 24 h on the plants, and remained higher after 7 d. Mites on cucumber also were more susceptible to the organophosphate dimethoate than were mites on lima bean, but only after 7 d on the host. Susceptibility was inversely related to activities of both general esterase and glutathione S-transferase (GST) in mites on cucumber; general esterase and GST activities were 60 and 25% lower, respectively, than activities of twospotted spider mite on lima bean after 7 d of feeding. Mites on maize were slightly but significantly more susceptible than those on lima bean to bifenthrin, but not to lambda-cyhalothrin, after 7 d and to dimethoate after 24 h but not after 7 d. General esterase and GST activities in twospotted spider mite fed on maize for 24 h were 20 and 16% higher, respectively, than activities in twospotted spider mite on lima bean, but general esterase activity was 30% lower than lima bean-fed mites and GST was not different after 7 d. Thus, plant-induced changes in general esterase activity, perhaps in combination with GST activity, in twospotted spider mite appear to be inversely related to, and possibly responsible for, changes in susceptibility of twospotted spider mite to several pesticides, particularly the synthetic pyrethroids. General esterases appear to play less of a role in the detoxification of the organophosphate insecticide dimethoate.     

Efficacy and persistence of rosemary oil as an acaricide against twospotted spider mite (Acari: Tetranychidae) on greenhouse tomato

Efficacy of rosemary, Rosmarinus officinalis L., essential oil was assessed against twospotted spider mite, Tetranychus urticae Koch (Acari: Tetranychidae), as well as effects on the tomato, Lycopersicum esculatum Mill., host plant and biocontrol agents. Laboratory bioassay results indicated that pure rosemary oil and EcoTrol (a rosemary oil-based pesticide) caused complete mortality of spider mites at concentrations that are not phytotoxic to the host plant. The predatory mite Phytoseiulus persimilis Athias-Henriot is less susceptible to rosemary oil and EcoTrol than twospotted spider mite both in the laboratory and the greenhouse. Rosemary oil repels spider mites and can affect oviposition behavior. Moreover, rosemary oil and rosemary oil-based pesticides are nonpersistent in the environment, and their lethal and sublethal effects fade within 1 or 2 d. EcoTrol is safe to tomato foliage, flowers, and fruit even at double the recommended label rate. A greenhouse trial indicated that a single application of EcoTrol at its recommended label rate could reduce a twospotted spider mite population by 52%. At that rate, EcoTrol did not cause any mortality in P. persimilis nor did it affect their eggs. In general, EcoTrol was found to be a suitable option for small-scale integrated pest management programs for controlling twospotted spider mites on greenhouse tomato plants.     

Comparative toxic and sublethal effects of fluvalinate on two-spotted spider mite and European red mite

Four fluvalinate formulations differed in their residual toxicity to female two-spotted spider mite (TSM), Tetranychus urticae adults; the emulsifiable concentrate (EC) was the most toxic. In contrast, there was little difference in toxicity between the formulations with the European red mite (ERM) Panonychus ulmi with the exception of the EC formulation which was the least toxic. Fluvalinate 2F caused minimal (<10%) TSM and ERM egg mortality. Fluvalinate 2F was more toxic and caused greater larval dispersal for the TSM compared to the ERM at the field concentration and below. The toxicity of fluvalinate 2F to TSM and ERM protonymphs, deutonymphs and adults was low, approximately <20% at field concentration. Dispersal was the main response to fluvalinate and this was positively correlated with increasing concentration. The combined mortality and dispersal LC₅₀ was five times lower for ERM protonymphs and adults, but 11 times higher for ERM deutonymphs compared to equivalent TSM life stages. Fluvalinate 2F reduced TSM development from the protonymph and deutonymph stages to a greater extent compared to the ERM. The mortality response to fluvalinate 2F was unaffected by host type (peach or apple) for the TSM whereas ERM mortality was higher on apple compared to peach. TSM dispersal was higher from apple compared to peach whereas ERM dispersal was similar on both host types. Oviposition by both mite species was lower on apple than peach leaves. A 1 h exposure to fluvalinate 2F reduced ERM oviposition for 12 days.     

Acaricidal activity and sublethal effects of an oxymatrine-based biopesticide on two-spotted spider mite (Acari: Tetranychidae)

Lethal and sublethal effects of the biopesticide Kingbo (oxymatrine 0.2 % + psoralen 0.4 %) on the two-spotted spider mite (Tetranychus urticae Koch) were investigated in laboratory bioassays. The biopesticide was applied to bean leaf discs or primary leaves by using a Potter spray tower. Acute toxicity tests showed no significant ovicidal action: toxic effect (LC₅₀ = 55.49 μl/l) was the result of a residual activity against larvae that hatched from the treated eggs. Preovipositional females and female teleiochrysales showed similar susceptibility (LC₅₀ = 52.68 and 59.03 μl/l, respectively), whereas larvae, protonymphs and female deutonymphs were the most susceptible stages (LC₅₀ = 6.88, 13.03, and 8.80 μl/l, respectively). In a choice test, females preferred the untreated halves of leaves over the halves treated with 2,000, 1,000, and 500 μl/l in the first 24 h, and their oviposition in those treatments was significantly greater on the untreated halves after 24 and 48 h, as well as the summed oviposition over 72 h. Viability and reproduction of survivors, as well as population growth, were strongly affected after the treatments of preovipositional females and female teleiochrysales with 100, 50 and 25 μl/l. On the other hand, sublethal effects on the females that survived treatment at the egg stage or reached adulthood from the eggs laid on the treated surface (treatments with 50 and 25 μl/l) were significantly weaker. Acaricidal and sublethal effects of the biopesticide Kingbo were discussed as a starting point for further research aimed to improve management of T. urticae populations. Regulatory issues and safety concerns regarding further commercialization of this biopesticide are addressed as well.     

Linkage between one of the polygenic hexythiazox resistance genes and an etoxazole resistance gene in the twospotted spider mite (Acari: Tetranychidae)

Genetic linkage between hexythiazox and etoxazole resistance loci was analyzed by crossing experiments. Two strains, one resistant (R) and the other susceptible (S) to both chemicals were established from field-collected Tetranychus urticae Koch (Acari: Tetranychidae) populations that were further selected in the laboratory. To analyze the recombination rate of the loci associated with resistance, we tested the ovicidal effects of a mixed solution of hexythiazox and etoxazole on haploid F2 eggs laid by F1 females from an R female x S male cross. This revealed tight or complete linkage between the hexythiazox and etoxazole resistance loci. We then assessed the number of loci associated with resistance to each acaricide based on mortality in the haploid F3 progeny (eggs) of F2 females from an F1 female (R x S) x S male testcross. The mortality rate indicated that etoxazole resistance was largely controlled by a single major locus, whereas hexythiazox resistance was controlled by more than one locus. Thus, one hexythiazox resistance locus was tightly or completely linked to the etoxazole resistance locus.     

Transovarial toxicity matters: lethal and sublethal effects of hexythiazox on the two-spotted spider mite (Acari: Tetranychidae)

The effects of hexythiazox on life-history traits and demographic parameters of Tetranychus urticae Koch (Acari: Tetranychidae) were evaluated using the age-stage two-sex life table (in fecundity-based and fertility-based variants), with emphasis on its transovarial toxicity. Hexythiazox was applied when T. urticae females were either in the preovipositional period or in the first day of oviposition. In the F₀ generation bioassay, treatments with concentrations of 50, 12.5 and 3.125 mg/l significantly reduced the longevity of females and their fecundity. These effects were mostly the result of mortality of treated females (18–23%) over the 24-h exposure period. Even though the net reproductive rate (R₀) decreased significantly, the intrinsic rate of increase (r), finite rate of increase (λ) and doubling time (D) were not significantly different from the control. The strongest transovarial toxic effect occurred within the first 4 days following treatment, when 52–89% of the eggs laid by treated females (96% in control) hatched. Fertility was significantly reduced by concentrations of 50, 12.5, 3.125, 0.781 and 0.195 mg/l. These concentrations caused significant reductions in R₀ (34–54%), r (12–24%) and λ (3–5%), whereas D was extended for 0.4–0.7 days. In the F₁ generation bioassay, 50, 12.5, 3.125, 0.781, 0.049 and 0.012 mg/l caused significant reductions in R₀ (34–92%), r (10–68%) and λ (3–17%), whereas extending D for 0.3–5.6 days. These effects were mostly the consequence of transovarial toxicity. Application of the fecundity-based life table underestimated population-level effects of hexythiazox on T. urticae.

     

Functional response of Neoseiulus barkeri Hughes on two-spotted spider mite (Acari: Tetranychidae)

Generalist predatory mites are the common phytoseiid fauna in many agroecosystems, but little attention has been paid to their potential as biological control agents. In this study, we determined the functional responses of adult females of the generalist predator Neoseiulus barkeri Hughes on eggs, larvae, and adults of the two-spotted spider mite, Tetranychus urticae Koch, in the laboratory. Predation experiments were conducted on pepper leaf discs over a 24 h period at 25±1°C, 70–80% RH and 16L:8D photoperiod. Prey densities ranged 5 to 80 eggs, or 5 to 40 larvae, or 1 to 8 female adults of T. urticae per disc. The predation rate of N. barkeri adult females on T. urticae eggs was the same as on its larvae, but the predation rate on adult females was much lower. The role of generalist predatory mites in integrated and biological control of greenhouse pests was discussed.     

Wolbachia和Cardinium对皮氏叶螨生殖的影响及在寄主体内的定位

Wolbachia和Cardinium均为母系遗传的胞内共生菌, 它们能够通过诱导胞质不亲和（cytoplasmic incompatibility, CI）以调控寄主的生殖。目前, 关于Wolbachia和Cardinium共同对同一寄主进行生殖操控的机制还不清楚。本研究以皮氏叶螨Tetranychus piercei McGregor广州种群为实验材料, 通过杂交实验和荧光原位杂交的方法, 研究Wolbachia和Cardinium单感染和双感染对寄主生殖的影响。结果表明： 单感染Wolbachia诱导较弱的CI, 不亲和组合的未孵化率为17.8%±1.6%。单感染Cardinium及双感染Wolbachia和Cardinium能诱导高强度的CI, 不亲和组合的未孵化率分别为70.3%±1.3%和72.9%±1.2%。同时双感染Wolbachia和Cardinium雌螨的平均产卵量为35.2±1.2, 显著高于单感染和不感染的雌螨的产卵量。Wolbachia 和Cardinium分别诱导以及共同诱导CI的水平与精子形成过程中的感染情况有关。Wolbachia和Cardinium的垂直传播模式结果显示, 在卵的不同发育阶段, Wolbachia和Cardinium主要伴随着营养物质从滋养细胞、 中肠、 输卵管进入发育中的卵。研究结果为进一步了解 Wolbachia和Cardinium的母系遗传机制提供了重要依据。     

The effect of interspecific mating on sex ratios in the twospotted spider mite and the Banks grass mite (Acarina: Tetranychidae)

Mixed populations of the twospotted spider mite (TSM),Tetranychus urticae (Koch), and the Banks grass mite (BGM),Oligonychus pratensis (Banks), occur on corn and sorghum plants in late summer in the Great Plains. Interspecific matings between these arrhenotokous species occur readily in the laboratory but yield no female offspring. The effect of interspecific mating on female: male sex ratios was measured by examining the F₁ progeny of females that mated with both heterospecific and conspecific males in no-choice situations. TSM females that mated first with BGM males and then with TSM males produced a smaller percentage of female offspring than TSM females that mated only with TSM males (43.1±5.8 and 78.9±2.8% females, respectively). Similarly, BGM females mated with heterospecific males and then with conspecific males produced fewer female offspring than females mated only with BGM males (55.7±5.2 and 77.5±2.5%, respectively). Lower female: male sex ratios were produced also by BGM females that mated with TSM males after first mating with conspecifics (62.4±3.4%). In mixed populations containing males of both species, females also produced lower female: male sex ratios, but these ratios were not as low as expected based on mating propensities and progeny sex ratios observed in no-choice tests. These data suggest that interspecific mating may substantially reduce female fitness in both mite species by reducing the output of female offspring, but in mixed populations this effect is mitigated by unidentified behavioral mechanisms.