Multifactorial resistance to transpermethrin in field-collected strains of the tobacco budworm Heliothis virescens F.

The penetration, degradation and excretion of [³H]transpermethrin were examined in susceptible and field-collected pyrethroid-resistant strains of the tobacco budworm Heliothis virescens. No consistent differences in labelled materials excreted or recovered in cuticle rinses were found between the resistant (R) and susceptible (S) larvae. Considerably lower levels of the parent compound were present internally in R compared with S larvae after 24h (P <0.01), clearly identifying a metabolic resistance mechanism in Meloland and Westmorland larvae. Moderate levels of absorbed permethrin accompanied by an absence of poisoning symptoms were observed in certain individuals of both R strains, suggesting a second resistance mechanism. Neurobioassays of R larvae showed a consistently lower sensitivity of the neuromuscular system to pyrethroids when compared with the S larvae, thus confirming the indication from metabolic studies of the additional (site-insensitive) mechanism. Toxicity values suggest a cross-resistance to other pyrethroids.     

Nerve insensitivity resistance to cypermethrin in larvae of the tobacco budworm Heliothis virescens from USA cotton field populations

Spontaneous neuronal activity was recorded from the peripheral nerves of third-instar larvae of strains of Heliothis virescens (F.) obtained directly from cotton fields in the USA. Following a control period the preparations were exposed to increasing concentrations of cis-cypermethrin in a cumulative dose-response assay. A positive response was defined as an increase of at least five-fold in the rate of neuronal activity over that seen during the control period. Up to 35 individuals of each strain were assayed and the responses used to construct a phenotypic profile categorising the individuals from nerve-susceptible to highly nerve-insensitive. An EC₅₀ for the action of cis-cypermethrin was also obtained. There was a positive, significant correlation between non-synergisable resistance to cypermethrin and nerve insensitivity as defined in the neurophysiological assay. It was shown that nerve insensitivity to cypermethrin increased throughout the cotton-growing season.     

Influence of oil droplet size on the toxicity of bifenthrin on cotton to tobacco budworm (Heliothis virescens)

Sprays of bifenthrin and paraffinic oil mixtures were applied to cotton leaves (Gossypium hirsutum L.) to investigate droplet size effects on the mortality of Heliothis virescens (F.). An air-assisted nozzle applied mixtures at 9.4 litre ha ?1 spray rate with volume median diameters (VMD) of 96 and 337 μm, as verified with a Malvern laser diffraction analyzer. An hour after spraying, susceptible 3rd-instars walked for 2-min through wet droplets of spray mixtures applied at bifenthrin rates of 22, 45, 67, 90. and 112 g a.i. ha ?1. Mortality was assessed at 21, 26, 45, 50, 74, and 117h after exposure. Final mean mortality was affected (P =0.01) by bifenthrin rate and droplet size. Bifenthrin applied in 337 μm VMD droplets caused 75.8% mortality, whereas 96-μm VMD droplets resulted in a mortality of 67.9%. Also, the 337 μm VMD droplets killed larvae in 13% less time than the 96μm VMD droplets, based on analysis (P =0.01) using the mean elapsed time till death as the dependent factor.     

Biological effects and metabolism of fenoxycarb after treatment of the fourth and the fifth instars of the tobacco budworm,Heliothis virescens F.

Treatment of 4th and 5th instar larvae of Heliothis virescens with different amounts of fenoxycarb induced an increase in weight of the 5th larval instar, prolongation of the 5th larval instar period, or the formation of dauer larvae. No effects were observed during the 4th instar and the moult to the 5th larval instar. To check whether these effects were due to persistence of fenoxycarb in treated larvae, metabolism of this chemical was analysed. Larvae were fed with radio-labelled fenoxycarb at the first day of the 4th or at the first day of the 5th larval instar. Behaviour of the radio-labelled compounds was followed during each instar. At the time of gut purge, the whole of the radio-labelled material was excreted. The effects observed during the 5th instar of 4th-instar-treated larvae do not appear to be due to the persistence of fenoxycarb. The main metabolites of fenoxycarb were identified using coupled gas chromatography and mass spectrometry.     

Interaction of proteinase inhibitors with Cry1Ac toxicity and the presence of 15 chymotrypsin cDNAs in the midgut of the tobacco budworm, Heliothis virescens (F.) (Lepidoptera: Noctuidae)

BACKGROUND: The potential development of resistance to Bacillus thuringiensis (Bt) cotton and surging of non‐targeted insects is a major risk in the durability of Bt plant technology. Midgut proteinases are involved in Bt activation and degradation. Proteinase inhibitors may be used to control a wide range of insects and delay Bt resistance development. Proactive action to examine proteinase inhibitors for synergistic interaction with Bt toxin and cloning of proteinase cDNAs for RNAi is necessary to make transgenic cotton more versatile and durable. RESULTS: A sublethal dose (15 ppb) of Cry1Ac, 0.5% benzamidine and 0.02% phenylmethylsulfonyl fluoride significantly suppressed midgut azocaseinase, tryptic and chymotryptic activities, and resulted in reductions in larval and pupal length and mass of Heliothis virescens. The combination of proteinase inhibitor and Bt suppressed 20–37% more larval body mass and 26–80% more enzymatic activities than the inhibitor only or Bt only. To facilitate knockdown‐resistance‐related proteinase genes, 15 midgut chymotrypsin cDNAs were sequenced. Most predicted chymotrypsins contained the conserved N‐termini IVGG, three catalytic center residues (His, Asp and Ser), substrate specificity determinant (Ser or Gly) and cysteines for disulfide bridges. These putative chymotrypsins were separated into three distinct groups, indicating the diverse proteinases evolved in this polyphagous insect. CONCLUSION: H. virescens has evolved diverse midgut proteinase genes. Proteinase inhibitors have potential insecticidal activity, and the interaction of Bt with proteinase inhibitors is desirable for enhancing Bt toxicity and delaying resistance development. Intensive sequencing of chymotrypsin cDNAs will facilitate future functional examinations of individual roles in Bt toxicity and resistance development and facilitate targeted control using RNAi and/or proteinase inhibitors. Copyright © 2012 Society of Chemical Industry     

Comparison of insecticide absorption and detoxification in larvae of the bollworm,Heliothis zea,and the tobacco budworm,H. virescens

Rates and nature of metabolism of several insecticides were compared in larvae of the bollworm, Heliothis zea (Boddie), and the tobacco budworm, H virescens (F.). Both species are resistant to organochlorine compounds. In addition, H. virescens is resistant to carbamate and organophosphate insecticides. All test insecticides were metabolized by larvae of both species under in vivo conditions. In most instances, metabolism was more rapid in H. virescens larvae.A series of in vivo and in vitro assays implicated high levels of NADPH-dependent oxidative enzymes as a major metabolic factor in resistance. Other work demonstrated that H. virescens larvae absorbed insecticides at a slower rate than did H. zea larvae. High resistance to organophosphates present in H. virescens may be due to interaction of factors for increased rate of detoxification and decreased rate of absorption.     

Toxicity of chlordimeform and amitraz to the egyptian cotton leafworm (Spodoptera littoralis) and the tobacco budworm (Heliothis virescens)

The relative toxicity (μg a.i. g^?1 body wt) of the formamidine insecticide chlordimeform (CDM) and the triazapentadiene insecticide amitraz was examined in two species of noctuid moth Spodoptera littoralis and Heliothis virescens. When applied topically, there was an unexpected and marked difference in the toxicity of CDM base and its hydrochloride to adults of both species, the salt being appreciably more toxic. For H. virescens at least, this difference in toxicity could not be explained by differences in penetration. This trend was reversed for larval instars of S. littoralis; while there was relatively little difference in the toxicity of the base to adult and larval stages, the salt was at least 1000-fold more toxic to adults than to larvae. N¹-Demethylchlordimeform (DCDM) was the only metabolite of CDM to show biological activity against either species, but was much less toxic than the parent compound. Amitraz was far less toxic than either CDM or DCDM; like the CDM salt, it appeared to be more toxic to adult than larval stages of S. littoralis. Application of piperonyl butoxide significantly increased the toxicity of the CDM salt, DCDM and amitraz to adult H. virescens, the synergist being particularly effective with DCDM and amitraz. In contrast, piperonyl butoxide had no significant effect on the toxicity of DCDM, and slightly antagonised the toxicity of DCDM to fourth-instar larvae of S. littoralis.     

Fenoxycarb,an insect growth regulator with juvenile hormone activity: A candidate for Heliothis virescens (F.) control on cotton

Tobacco budworm eggs, deposited on cotton plants (Gossypium hirsutum L.) treated with fenoxycarb, do not produce viable larvae. The insects die either at the end of embryonic development or during 1st-2nd larval instars, even when the larvae hatching on the treated leaves are transferred to untreated plants. This ovo-larvicidal activity and indications of good foliar persistence make fenoxycarb a promising candidate for the control of tobacco budworm on cotton. This type of application could open new aspects for the control of lepidopterous pests with this novel compound.     

Acute toxicity of diazinon on the common carp (Cyprinus carpio L.) embryos and larvae

In the present study, the toxic effects on the embryos and larvae of the common carp were used as a model to investigate the organophosphorus pesticide, diazinon, which contaminates aquatic ecosystems. Data obtained from the diazinon acute toxicity tests were evaluated using the Probit Analysis Statistical Method. The control and six test experiments were repeated five times. The number of dead embryos significantly increased in response to diazinon concentrations 0.25, 0.5, 1, 2, 4, and 8 mg L⁻¹ (p < 0.05 for each case). The 48 h LC₅₀ value (with 95% confidence limits) of diazinon for common carp embryos was estimated at 0.999 (0.698-1.427) mg L⁻¹. Dose-response decreases in hatching success were recorded as 84.60, 75.2, 54.1, 31.0, 6.0, and 0.0%, respectively (p < 0.05). The number of dead larvae significantly increased with increasing diazinon concentrations exposed for 24-96 h (p < 0.05). The 24, 48, 72, and 96 h LC₅₀ values (with 95% confidence limits) of diazinon for common carp larvae were estimated at 3.688 (2.464-8.495), 2.903 (2.019-5.433), 2.358 (1.672-4.005), and 1.530 (1.009-3.948) mg L⁻¹, respectively. There were significant differences in the LC₅₀ values obtained at different exposure times (p < 0.05). The results of the study suggest that low levels (0.25 mg L⁻¹) of diazinon in the aquatic environment may have a significant effect on the reproduction and development of carp.     

Synergism of diazinon toxicity and inhibition of diazinon metabolism in the house fly by tridiphane: Inhibition of glutathione S-transferase activity

Diazinon toxicity to a susceptible strain of house fly (Musca domestica L.) was synergized by tridiphane [2-(3,5-dichlorophenyl)-2-(2,2,2-trichloroethyl)oxirane], a herbicide synergist. Both diazinon and tridiphane were partially metabolized in the house fly by glutathione (GSH) conjugation. Synergism appeared to be due to inhibition of diazinon metabolism/detoxification. Crude glutathione S-transferase (GST) preparations from the house fly catalyzed GSH conjugation of diazinon, tridiphane, 3,4-dichloronitrobenzene (DCNB), and chloro-2,4-dinitrobenzene (CDNB). Tridiphane and the GSH conjugate of tridiphane appeared to inhibit diazinon GSH conjugation, but diazinon did not inhibit tridiphane GSH conjugation. The enzymatic rate of tridiphane GSH conjugation was 22 times the rate of diazinon GSH conjugation; therefore, attempts to assay tridiphane as an inhibitor of diazinon GSH conjugation were inconclusive because of the high concentration of tridiphane GSH conjugate produced during the assay. CDNB underwent enzymatic GSH conjugation at a rate 240 times faster than that of tridiphane and 5000 times faster than that of diazinon. GSH conjugation of CDNB was not inhibited by tridiphane, but was inhibited by the GSH conjugate of tridiphane. In vivo, the GSH conjugate of tridiphane was produced in sufficient concentration to cause the observed inhibition of diazinon metabolism and synergism of diazinon toxicity. However, the possibility that parent tridiphane caused or contributed to the inhibition of diazinon metabolism and synergism of diazinon toxicity could not be excluded. Inhibition of diazinon metabolism did not appear to be due to depletion of either GSH or GST.     

烟芽夜蛾囊泡病毒3h株3h-122基因于棉铃虫幼虫体内的转录表达分析

本文构建了烟芽夜蛾囊泡病毒3h株(HvAV-3h)第122个基因(3h-122)的原核表达载体并制备多克隆抗体?利用蛋白免疫印迹?RT-PCR?免疫组织化学染色等技术检测3h-122的转录表达时相以及组织特异性?结果表明, 3h-122编码大小16.1 kD的结构蛋白(3H-122)?HvAV-3h感染棉铃虫幼虫后3 h开始检测到3h-122的转录并稳定持续至第168 h?3H-122在棉铃虫感毒后24 h至168 h可持续检测到其表达?此外, 3H-122主要在脂肪体组织中表达?本研究为阐明囊泡病毒结构蛋白的功能及其在囊泡病毒致病机制中的作用奠定了基础?     

Acephate toxicity,metabolism, and anticholinesterase activity in Heliothis virescens (F.) and Anthonomus grandis grandis (Boheman)

The toxicity (72 hr) of acephate and methamidophos to fourth-instar larvae of the tobacco budworm, Heliothis virescens (F.), was nearly equivalent. In contrast, toxicity (72 hr) of methamidophos to adult boll weevils, Anthonomus grandis grandis (Boheman), was substantially greater than that of acephate. The internal accumulation of acephate was greater for A. grandis grandis than for H. virescens at 24 and 48 hr post-treatment, as was excretion. Acephate was metabolized to methamidophos both in vivo and in vitro by H. virescens but not by A. grandis grandis. In vitro acetylcholinesterase inhibition by methamidophos was greater than that of acephate, but less than that of paraoxon for H. virescens, A. grandis grandis, and the electric eel. Treatment of H. virescens larvae with acephate resulted in increased in vivo acetylcholinesterase inhibition between 24 and 72 hr post-treatment, which was associated with a large increase in mortality. H. virescens treated with methamidophos showed greater mortality and greater acetylcholinesterase inhibition at earlier time periods than those treated with acephate. However, by 72 hr post-treatment, in vivo acetylcholinesterase inhibition by LD₅₀ doses of acephate and methamidophos were approximately equivalent. These results indicate that, for H. virescens, toxicity of acephate is directly related to its metabolism to methamidophos and subsequent acetylcholinesterase inhibition. Likewise, the differential toxicity of acephate and methamidophos to A. grandis grandis adults appears to be due to their inability to metabolize acephate to methamidophos.     

Host plant induction of glutathione S-transferase in the fall armyworm

The influence of various host plants on glutathione S-transferase activity was studied in the fall armyworm, Spodoptera frugiperda (J. E. Smith). Fall armyworm larvae were maintained on a semidefined artificial diet until the end of the fifth instar. The newly molted sixth instar larvae were then fed fresh leaves of various host plants for 2 days prior to glutathione S-transferase assays using 3,4-dichloronitrobenzene as substrate. The order of the midgut glutathione S-transferase activity of larvae after the worms fed on these plants was: mustard > turnip > cowpeas > peanuts > cotton > corn > cucumber > potato > Bermudagrass > millet > sorghum > soybeans. The difference in the transferase activity between soybean- and mustard-fed larvae was 10-fold. Kinetic study revealed a quantitative, but no qualitative difference in the glutathione S-transferase between soybean- and cowpea-fed larvae. Monoterpenes, such as α-pinene, β-pinene, menthol, and peppermint oil, had no effect on the enzyme. Cowpea-fed larvae were more tolerant of the insecticides diazinon, methamidophos, and methyl parathion than soybean-fed larvae were. These new observations help explain what has been happening in the field and might be of use in the development of pest management programs.     

The dietary toxicity of deltamethrin to the carabid,Nebria brevicollis (F.)

Batches of adult Nebria brevicollis (F.) (Coleoptera: Carabidae) were provided with freeze-killed aphids, Metopolophium dirhodum (Walk.) (Homoptera: Aphididae), that had been treated topically with 30 ng a.i. deltamethrin. Beetles in different hunger states were found to consume contaminated aphids, but they consumed significantly fewer aphids than control beetles. The mean numbers of contaminated aphids eaten increased with hunger-level, from 2.5 aphids per beetle for those provided with food 24 h before exposure, to 4.5 aphids per beetle for those provided with food 120 h prior to exposure. The highest mortality that occurred over the following six days after consumption of contaminated aphids was 27% for the beetles that had been fed 120 h before exposure and the lowest was 13% for the beetles that had been fed 24 and 48 h prior to exposure. A regurgitation response was observed in 53–80% of the beetles that had consumed deltamethrin-treated aphids. The beetles that survived consumption of treated aphids showed a short-term reduction in mean numbers of untreated aphids consumed relative to the control beetles. Dietary and topical dose-response data for deltamethrin and N. brevicollis were compared by probit analysis and maximum likelihood procedures. The dietary exposure probit line had a significantly steeper slope than the topical exposure probit line but the two lines occupied similar positions along the dose axis. The steepness of the dietary exposure probit line may be partly explained by the regurgitation response shown by the beetles, which causes water loss and may amplify toxic effects. The results indicated that the consumption of deltamethrin-contaminated prey may be an important cause of predator mortality soon after pesticide application.     

烟芽夜蛾囊泡病毒3h株对草地贪夜蛾幼虫的感染特性及对其生长发育的影响

为寻找有效防控外来入侵物种草地贪夜蛾Spodoptera frugiperda的技术,以烟芽夜蛾囊泡病毒3h株(Heliothis virescens ascovirus 3h,HvAV-3h)为研究对象,通过测定不同龄期草地贪夜蛾幼虫口服或针刺感染HvAV-3h后的死亡率、存活时间、取食量及体重,分析该毒株对草地贪夜蛾幼虫的感染特性及对其生长发育的影响。结果显示,草地贪夜蛾1、2龄幼虫口服HvAV-3h后的校正死亡率分别为9.22%和0,3～6龄幼虫针刺感染HvAV-3h后的校正死亡率则高达100.00%,感染HvAV-3h的幼虫均在幼虫期或蛹期死亡。3～5龄幼虫针刺感染HvAV-3h后的存活时间明显长于健康幼虫;3～6龄幼虫针刺感染HvAV-3h后其体重和取食量均受到不同程度的抑制作用,体重抑制率分别为67.79%、41.68%、16.31%和10.30%,总取食量抑制率分别为57.80%、33.90%、17.42%和41.82%;其中3龄幼虫针刺感染HvAV-3h后的体重和取食量被显著抑制,且蜕皮困难,最终在幼虫期死亡;部分4～6龄感染HvAV-3h幼虫能够完成化蛹,但是均无法羽化。表明HvAV-3h感染能够有效控制草地贪夜蛾幼虫,有望开发为草地贪夜蛾的生防制剂。     

Laboratory Studies on Host Preference and Insecticide Resistance of the Cotton Bollworm,Heliothis obsoleta F.1

Pupae of the cotton bollworm, Heliothis obsoleta F., from the northern part of Iran (Gorgan and Gonbad areas) were brought to the laboratory and used as the breeding source for bollworms. The larvae were reared on soybean plants for 8 generations under laboratory conditions of 60±5 % RH and 27±2 °C. To test host preference, the adults of both sexes were released into wooden cages where they were able to freely select their host. The females laid more eggs on soybean (before and at blooming) compared to cotton (before and after blooming) and maize (before and after stem extension). Seven and 14-day-old larvae were treated with different doses of Supracide and Thiodan by topical application. Altogether, 3,000 larvae were used for this experiment and from the data collected, LD 50 values were calculated. It was found that, as the age doubled from 7 to 14 days, the LD 50 value for Supracide increased from 0.6 mg/g to 0.92 mg/g. For Thiodan, the corresponding increase during this period was from 0.52 mg/g to 1.50 mg/g. Fat body content was measured using Soxhlet apparatus and its correlation with LD 50 values was determined. Fat bodies represented 2.1 and 2.5% of the body weight in 7 and 14-day-old larvae, respectively.     

Mechanism of resistance to spinosyn in the tobacco budworm, Heliothis virescens

Topical laboratory selection of tobacco budworm larvae, Heliothis virescens, with technical spinosad for multiple generations resulted in larvae 1068-fold resistant to topical applications of the insecticide and 316.6-fold resistant to insecticide treated diet as compared to the parental strain. The penetration of 2′-O-methyl[¹⁴C]spinosyn A across the cuticle of the susceptible (parental) and selected (resistant) tobacco budworms increased with time 3-12 h after application. A trend of reduced penetration in the resistant strain was found but the differences were not statistically significant. 2′-O-methyl[¹⁴C]spinosyn A when injected into the hemocoel was not metabolized 96 h after treatment in both the susceptible and resistant strain, suggesting that a change in metabolism was not the mechanism of resistance. Electrophysiological studies indicated that dose-dependent spinosyn A-induced currents occurred in neurons from spinosyn resistant and susceptible (adult) tobacco budworms. At both 10 and 100 nM spinosyn A, however, the amplitude of these currents in the resistant insects was significantly smaller than the amplitude of currents observed from neurons from susceptible tobacco budworm adults. This suggests that neurons from resistant insects have decreased sensitivity to spinosyn A. However, the reduced inward currents in the resistant strain may or may not be related to the mode of action of the spinosyns. No statistically significant cross-resistance was noted for the spinosad resistant tobacco budworms for topical applications of permethrin (Pounce^®), profenofos (Curacron^®), emamectin benzoate (Denim^®), or indoxacarb (Steward^®). A statistically significant reduction in susceptibility to acetamiprid (Mospilan^®) in artificial diet as determined from a resistance ratio of 0.482 was found.     

Xenobiotic metabolism of plant secondary compounds in the English grain aphid, Sitobion avenae (F.) (Hemiptera: Aphididae)

Plant secondary compounds have been documented to be deleterious to insects and other herbivores in diverse ways. In this study, the effect of catechol (phenolics), gramine (alkaloid) and L-ornithine-HCI (non-protein amino acid) on the activities of xenobiotic metabolizing enzymes in English grain aphid, Sitobion avenae, was evaluated. Phase I enzymes investigated in this study included carboxylesterase (CarE), and oxidoreductase, whereas Phase II enzymes were represented by glutathione S-transferase (GST). In general, CarE and GST activities in S. avenae were positively correlated with the concentration of plant secondary compounds in artificial diets. Oxidoreductase activity, however, displayed a different profile. Specifically, peroxidase (POD) and polyphenol oxidase (PPO) activities in S. avenae were positively correlated with concentrations of dietary catechol and gramine, respectively, whereas catalase (CAT) activity was significantly suppressed by the higher concentration of catechol, gramine and L-ornithine-HCl. These combined results suggest that CarE and GST in S. avenae are key enzymes to breakdown a broad spectrum of plant secondary compounds, whereas oxidoreductase, including PPO and POD, degrades specific groups of plant secondary compounds.     

Efficacy of spiromesifen on Aceria litchii (Keifer) in relation to Cephaleuros virescens Kunze colonization on leaves of litchi (Litchi chinensis Sonn.)

Journal of Plant Diseases and Protection - The phytophagous mite, Aceria litchii (Keifer), is one of the major pests of litchi (Litchi chinensis Sonn.) in Asia and Australia. By sucking on the...     

Definition of the flight period of Lasioderma serricorne (F.) in stored tobacco

A 4-year (1996–99) survey of the biology of L. serricorne (F) (Coleoptera, Anobiidae) on stored tobacco has taken place in Thessaloniki, northern Greece. For the trapping of the adults, pheromone ('New Serrico' and 'Black Stripe Moth') as well as electric (black light) traps were used. The first generation adults emerging from larvae hibernating inside the tobacco bales appeared flying on the 17/5, 16/5, 28/4, and 30/4 of the years 1996, 1997, 1998, and 1999, respectively. In parallel observations, adults inside the bales were found on the 14/5, 12/5, 23/4, and 26/4 of the above-mentioned years, respectively. Termination of flights, in all the years concerned, took place during October or November, while the last flights of November were recorded in temperatures of about 13°C.