Phenyltin compounds inhibit digestive enzymes of Tribolium confusum larvae

The potency of dietary phenyltin compounds in inhibiting the growth of first and fourth instar Tribolium confusum L. larvae and the gut proteolytic activity of fourth instar larvae decreases in the order of triphenyltin chloride (Ph₃SnCl) ? diphenyltin dichloride (Ph₂SnCl₂) ? phenyltin trichloride or tetraphenyltin. The growth retardation, which prolongs the larval stage without affecting pupation or emergence, may result from an antifeeding effect involving gut protease inhibition by Ph₃Sn⁺ and Ph₂Sn²⁺. Gut amylase and invertase activities are less sensitive than the protease activity to in vivo inhibition. Under in vitro conditions, relatively high concentrations of Ph₃SnCl and Ph₂SnCl₂ are required for inhibition, the order of enzyme sensitivity is protease > amylase > invertase, and Ph₂SnCl₂ is more potent than Ph₃SnCl. Proteins such as casein, albumin and hemoglobin, but not carbohydrates such as starch and sucrose bind Ph₃Sn⁺ so it is inaccessible for inhibition of digestive enzymes. The level of Ph₃Sn⁺ inhibiting gut protease in vivo is far below that necessary for in vitro inhibition of this enzyme activity. It is speculated that the in vivo inhibitory effects of Ph₃Sn⁺ and Ph₂Sn²⁺ on digestive enzymes may result from binding to the enzyme protein, its zymogen or to other proteins involved in production of the digestive enzymes.     

Triphenyl derivatives of group IV elements as inhibitors of growth and digestive enzymes of Tribolium castaneum larvae

The potency of triphenyl derivatives of group IV elements in inhibiting the growth of first and fourth instar Tribolium castaneum larvae is much greater for Ph₃SnCl and Ph₃PbCl than for Ph₃GeCl, Ph₃SiOH, and Ph₃CCl. Pupated larvae emerge normally, showing that the pupal stage is not affected. The larval growth retardation may result from an antifeeding effect involving digestive enzyme inhibition. Ph₃SnCl and Ph₃PbCl at 500 μmol/kg of diet completely inhibit larval growth (fourth instar larvae) and provide a reduction of 56–59, 29–33, and 2–15% in the in vivo activity of invertase, amylase, and protease, respectively. Under these conditions Ph₃GeCl, Ph₃SiOH, and Ph₃CCl are essentially inactive. High concentrations (5 × 10^?4 and 2 × 10^?3M) of Ph₃SnCl and Ph₃GeCl acting in vitro strongly inhibit invertase, amylase, and protease activities, whereas Ph₃PbCl is moderately inhibitory and Ph₃SiOH and Ph₃CCl are inactive. When both in vivo and in vitro findings are considered, Ph₃SnCl is the most potent inhibitor of larval digestive enzymes.     

Properties of Tribolium gut chitin synthetase

An insect chitin synthetase (CS) is readily assayed using the microsomal fraction (～0.5 mg protein) from an homogenate of Tribolium castaneum larvae. This enzyme preparation is incubated at 22°C with uridine 5′-diphospho-N-acetyl[³H]glucosamine in 355 μl of 25 mM Tris-HCl buffer containing 10 mM MgCl₂, 17 mM N-acetylglucosamine, and 1 mM dithiothreitol. Other divalent cations and amino sugars are less effective activators or are inhibitory. T. castaneum CS is strongly inhibited by polyoxin D and uridine 5′-diphosphate. These activation and inhibition properties of Tribolium castaneum gut CS are similar to those of fungal CS. The polymerization product formed by the Tribolium enzyme is stable in alkali but hydrolyzed by chitinase. Enzymes of Tribolium confusum, Tribolium brevicornis, Tenebrio molitor, and Galleria mellonella are also active under the same conditions. These enzymes are from the gut and probably from the peritrophic membrane. Integumental CS activity is not detected under the indicated assay conditions.     

Inhibition of Tribolium gut chitin synthetase

The chitin synthetase (CS) of Tribolium castaneum gut is inhibited 50% by 0.02 μM nikkomycin and 4 μM polyoxin D, two pyrimidine nucleoside fungicides, in in vitro assays with 10-min preincubation of enzyme and inhibitor prior to substrate addition. Tribolium CS is also sensitive to inhibition by the pyrimidine nucleotides uridine and cytidine di- and triphosphates. Captan, a known inhibitor of insect chitin synthesis, and the related fungicides captafol and dichlofluanid are highly potent inhibitors of Tribolium CS. Moderately active CS inhibitors are the acaricide oxythioquinox and the herbicide barban. One phenylcarbamate insect growth reatardant, H-24108, is weakly active in inhibiting Tribolium gut CS, as are three of its analogs but not 26 others. Many triazines are not inhibitory including several herbicides and an azido derivative, CGA 19255, which is active in blocking insect growth and chitin synthesis. Although the benzoylphenyl urea insecticides diflubenzuron and SIR 8514 are potent in vivo inhibitors of the polymerization step in insect chitin synthesis, they do not affect T. castaneum gut CS activity in vitro and greatly stimulate Tribolium brevicornis gut CS activity in vivo. These studies and preliminary findings on an integumental enzyme indicate that CS of these tissues is not sensitive to the direct action of benzoylphenyl ureas. This leads to speculation that the benzoylphenyl ureas act either as CS inhibitors via active metabolites formed in the integument or as blocking agents by direct binding to non-CS sites important in chitin polymerization and fibrillogenesis.     

Synthetic pyrethroids: Toxicity and synergism on dietary exposure of Tribolium castaneum (Herbst) larvae

The potency of six dietary pyrethroids, as toxicants and inhibitors of weight gain in first- and fourth-instar Tribolium castaneum (Herbst) larvae, decreased in the order of cis-cypermethrin and deltamethrin > trans-cypermethrin and cis-permethrin > fenvalerate and trans-permethrin. Dosages that reduced larval weight also delayed pupation and emergence, probably due to their antifeeding activity. Three oxidase inhibitors (piperonyl butoxide, O, O-diethyl O-phenyl phosphorothioate, and O-isobutyl O-prop-2-ynyl phenylphosphonate), at a dietary concentration of 100 mg kg^?1, had little or no effect on the toxicity of trans-permethrin, but strongly synergised the toxicity of cis-cypermethrin by about 3-, 3- and 10-fold, respectively. Piperonyl butoxide also synergised the toxicity of cis-permethrin, trans-cypermethrin and deltamethrin, but not that of fenvalerate. On the other hand, an esterase inhibitor, profenofos, did not enhance the potency of any of the α-cyano-3-phenoxybenzyl pyrethroids. Oxidases appear to be more important than esterases in pyrethroid detoxification by T. castaneum larvae.     

The effect of IKI-7899, a new chitin synthesis inhibitor,on larvae ofTribolium castaneum andSpodoptera littoralis

The effect of IKI-7899 and diflubenzuron on larval development and emergence ofTribolium castaneum and ofSpodoptera littoralis was determined. The LC₅₀ of IKI-7899 at adult emergence ofT. castaneum larvae fed the compound in their diet and the ED₅₀ of S.littoralis larvae treated topically were approximately one-sixth those of diflubenzuron.     

Effect of diflubenzuron on growth and carbohydrate hydrolases ofTribolium castaneum

The potency of diflubenzuron is much greater in inhibiting growth and development of 1st instar larvae ofTribolium castaneum than of 4th instar larvae, as expressed by death at the apolytic stage and retardation of larval development. A dose-dependent decrease in the activity of trehalase, invertase and amylase was obtainedin vivo with the increase in diflubenzuron concentration. At 5 ppm dietary concentration, a reduction of 37 and 27% in invertase and trehalase activity, respectively, was obtained in 4th instar larvae fed for 3 days on treated diet. The amylase activity was affected to a lesser extent. The observed disturbances of trehalase activity might hamper the supply of glucose needed for chitin build-up and those of invertase and amylase activity might affect feeding. Diflubenzuron does not inhibit these enzymesin vitro; hence, thein vivo effect seems to result from general disturbances in carbohydrate metabolism.     

低剂量甲氧虫酰肼对舞毒蛾生长发育的影响

采用药剂短期处理和连续处理,测定了低剂量LC_(10)和LC_(30)甲氧虫酰肼处理舞毒蛾4龄幼虫后,对其各项发育指标、营养指标及各发育阶段形态的影响.结果表明,LC_(10)和LC_(30)剂量甲氧虫酰肼能显著降低舞毒蛾幼虫成活率、化蛹率、6龄虫重及雌蛹重,与对照组相比,LC_(30)剂量处理24 h后,舞毒蛾5龄和6龄幼虫成活率、化蛹率、6龄虫重及雌蛹重明显降低,依次为48.89%、72.62%、62.22%、0.21 g和0.75 g,且LC_(30)剂量对幼虫和蛹的致死性较LC_(10)剂量明显,连续处理较短期处理明显,差异均显著(P<0.05).低剂量甲氧虫酰肼对幼虫营养指标也存在明显影响,LC_(10)剂量能抑制幼虫相对取食量(RCR),LC_(30)剂量能抑制食物利用率(ECI),且两者均能显著抑制幼虫相对生长率(RGR)(P<0.05).同时低剂量甲氧虫酰肼能导致舞毒蛾幼虫畸形,畸形率为62.53%.说明低剂量甲氧虫酰肼可取代高毒农药,用于防治森林鳞翅目害虫.     

Induction of prolonged larval feeding stage by juvenile hormone analogues inTribolium Castaneum

Dietary ZR-512 and ZR-619 at concentrations of 10 — 1000 ppm induced prolongation of the larval feeding period up to tenfold, increasing larval weight up to double that of untreated larvae. A comparison study of four juvenoids, using 200 ppm of ZR-512, ZR-515, ZR-619 or ZR-777, showed that ZR-515 elicits the highest larval weight (6.2 mg) and ZR-777 the lowest (3.6 mg). In all cases a pronounced enhancement of larval weight — of 50 — 250% relative to untreated larvae (2.4 mg) — was obtained.Tr. castaneum larvae reared up to their 3rd instars on a diet containing 100 ppm of ZR-512, ZR-515, ZR-619 or ZR-777 and then transferred to a juvenile hormone-free diet, were not affected. The period between 4th instar larva and pupation should therefore be considered as critical for juvenile hormone effect. The induced prolongation of the larval stage after juvenile hormone treatment was followed by a pronounced enhancement of cuticle phenoloxidase activity, indicating an alteration of the larval biochemical processes. Although juvenile hormone treatment inhibitsTr. castaneum pupation and emergence, it markedly prolongs larval feeding stage and weight and thus accelerates damage.     

Postdiapause development and spring emergence of the european corn borer,Ostrinia nubilalis,in Israel

The European corn borer (ECB),Ostrinia nubilalis (Hübner) (Lepidoptera: Pyralidae), is the major pest of corn (Zea mays L.) in Israel. We investigated the temporal patterns of pupation and emergence of various overwintering ECB populations in Israel during 1992-94. The association between Julian date (JD) or cumulative degree-days (DD) and the rates of either pupation or emergence was studied using simple linear regression models. Differences between populations in JDs required to reach 50% pupation amounted to 5%, whereas for DD differences amounted to 26%. Similarly, at 50% emergence, differences between populations were up to 3% for JD and to 12% for DD. Two different forecasting models are proposed for either pupation or emergence. Based on these models, both pupation and emergence develop over a period of 4 to 5 weeks, and they are expected to occur between the following JDs: onset of pupation, 66–70 (March 7–11); 50% pupation, 96–102 (April 6–12); onset of emergence, 85–94 (March 26-April 4); and 50% emergence, 115–121 (April 25-May 1). Pupal development required 160 DD (confidence interval [C.I.] 141–179 DD) and it is expected to takeca 16 days (C.I. 14–18 days). The simple linear regression models obtained in this study are suggested as preliminary phenological models for the temporal prediction of postdiapause pupation and emergence of ECB.     

Dietary triorganotins affect lymphatic tissues and blood composition of mice

Ph₃SnCl, Ph₃SnAc (Brestan), Cy₃SnOH (Plictran), and (Bu₃Sn)₂O (TBTO) markedly reduce the weight of the spleen and the gain in body weight of young mice feeding for 7 days on diets containing 260 μequiv organotin/kg diet. With mature mice feeding for 4 days on diets containing 780 μequiv organotin/kg, these compounds result in a loss in spleen weight and body weight and an alteration in the blood composition (reduction in lymphocytes and total leucocytes and an increase in erythrocytes, hemoglobin level, and hematocrit value). Four other organotins, [(PhCMe₂CH₂)₃Sn]₂O (Vendex), PhSnCl₂, Ph₂SnCl₂, and Ph₄Sn, are much less toxic based on these parameters. Poisoning by these dietary triorganotins was not related to cerebral edema or alterations in adrenal epinephrine level, liver nonprotein -SH groups, or the activity of digestive enzymes (intestine and stomach proteases and intestine amylase and invertase). The most sensitive indices of triorganotin toxicity appear to be those associated with changes in the lymphatic tissues and blood composition.     

Interaction of Cry1Ac toxin (Bacillus thuringiensis) and proteinase inhibitors on the growth, development, and midgut proteinase activities of the bollworm, Helicoverpa zea

Potential resistance development to Bt cotton in certain lepidopterans has prompted research to develop strategies that will preserve this environmental-friendly biotechnology. Proteinase inhibitors are potential candidates for enhancing Bt toxicity against lepidopteran pests and for expanding the spectrum of control for other insects. Interactions of Bt toxin from Bacillus thuringiensis and proteinase inhibitors were investigated by monitoring growth, development, and gut proteinase activities of the bollworm, Helicoverpa zea. Several proteinase inhibitors were combined with Bt protoxin Cry1Ac in artificial diet and fed to newly molted 3rd-instar bollworm larvae to determine effects on larval body weight and length, pupation progress, and mortality rate. Major midgut proteinase activities, including caseinase, tryptic, and chymotrypsin activities, were examined after treatment. A concentration of Bt at a level causing minimal mortality (<10%), was mixed with the following proteinase inhibitors: benzamidine, phenylmethylsulfonyl fluoride (PMSF), and N-α-tosyl-l-lysine chloromethyl ketone (TLCK). When compared with controls, the synergistic effect of Bt toxin and proteinase inhibitors caused significant decreases in mean larval weight and length over time. Midgut samples tested against the substrates azocasein, α-benzoyl-dl-arginine-p-nitroanilide (BApNA), and N-succinyl-alanine-alanine-proline-phenylalanine-p-nitroanilide (SAAPFpNA) showed significant decreases in the protease activity of larvae fed Bt plus inhibitor versus control. Interaction of Bt and proteinase inhibitors significantly retarded larval growth and resulted in developmental delay and up to 20% mortality.     

Toxicity nanoinsecticide based on clove essential oil against Tribolium castaneum (Herbst)

This study aimed to characterize nanoparticles loaded with clove (Syzygium aromaticum) essential oil-based polyethylene glycol (PEG) and to know their insecticidal activity against red flour beetle (Tribolium castaneum). The nanoparticles have irregular shapes in good dispersion. The nanoformulation could not enhance clove oil contact toxicity to T. castaneum, but could protect the oil from degradation and evaporation while simultaneously allowing sustained release, as indicated by the continued high toxicity for 16 weeks of storage.     

四种低毒杀虫剂对小菜蛾生长发育及繁殖的亚致死效应

为系统评价4种低毒杀虫剂对小菜蛾Plutella xylostella（L.）的综合控制作用,采用叶片浸渍法处理小菜蛾3龄幼虫,测定了虫酰肼、氟啶脲、茚虫威和虫螨腈的亚致死浓度（LC₂₅）对小菜蛾生长发育和繁殖力的影响。结果表明,4 种药剂亚致死浓度处理使小菜蛾3~4龄幼虫发育历期延长0.43~1.93 d,平均单头雌、雄蛹重分别减轻0.83~1.31 mg和0.19~1.09 mg ,幼虫的化蛹率及羽化率则分别降低11.2%~34.3%和21.7%~33.6%,以氟啶脲和茚虫威处理的影响较为显著。药剂处理后成虫平均产卵量减少30.15~60.21粒/雌,卵孵化率降低8.85%~19.65%,以虫酰肼效果较为明显;虫酰肼、氟啶脲、茚虫威均显著降低小菜蛾的交配成功率。     

噻虫胺等药剂对韭菜迟眼蕈蚊的致毒效应

为了明确新烟碱类药剂对韭菜迟眼蕈蚊的毒力,采用管测药膜法和药液定量滴加法测定了噻虫胺等6种药剂对韭菜迟眼蕈蚊不同虫态的毒力,并研究了噻虫胺、甲氨基阿维菌素苯甲酸盐和辛硫磷亚致死浓度对其4龄幼虫生长发育和繁殖的影响。结果表明,新烟碱类杀虫剂噻虫胺、吡虫啉和噻虫嗪对成虫的击倒毒力均较高,分别是阿维菌素的5.75、3.86和3.51倍;6种药剂对韭菜迟眼蕈蚊卵的毒力均较低;对2龄和4龄幼虫的毒力,均以噻虫胺最高,LC₅₀分别为0.339 mg/L和1.020 mg/L,分别是阿维菌素的27.00倍和25.23倍。用噻虫胺亚致死剂量处理韭菜迟眼蕈蚊4龄幼虫,其发育历期和蛹期延长,蛹重、化蛹率、成虫羽化率、单雌产卵量和卵孵化率均降低。     

Effects of interactions among Metarhizium anisopliae, Bacillus thuringiensis and chlorantraniliprole on the mortality and pupation of six geographically distinct Helicoverpa armigera field populations

A local isolate of Metarhizium anisopliae (Hypocreales: Clavicipitaceae), Bacillus thuringiensis subsp. kurstaki and chlorantraniliprole were assessed against six field populations of tomato fruitworm Helicoverpa armigera Hübner (Lepidoptera: Noctuidae) in a series of laboratory bioassays. Two dose rates of B. thuringiensis (0.5, 1 μg g^?1), one of both M. anisopliae (1.3?×?10⁶ conidia ml^?1) and chlorantraniliprole (0.01 ppm) were applied alone and in combination with each other against 2nd, 3rd, 4th and 5th larval instars. The mortality was observed every 24 h until pupation. The bioassays were carried out at 25°C and 75% r.h. The highest mortality was observed in Rawalpindi with the lowest pupation rate by applying the combined concentrations of B. thuringiensis and chlorantraniliprole. The lowest mortality was observed in population from Gujranwala among all the tested populations. The antagonistic interaction was noted where the high dose rate of B. thuringiensis was combined with M. anisopliae; however, the remaining interactions enhanced the mortality and reduced the percent pupation. The overall results demonstrated that all the treatments gave significant control of the larval instars of H. armigera. The population from Gujranwala proved least susceptible whereas the one from Rawalpindi was highly susceptible.     

Noxious effects of neem extracts onCrocidolomia binotalis

The potential of neem extracts to control the cabbage webworm,Crocidolomia binotalis Zell. (Lepidoptera: Pyralidae), was investigated in the laboratory. Neem, besides being an antifeedant at as low as 0.001% of the methanolic extract, was shown to be very toxic, as evidenced by high larval mortality and poor emergence. It also caused disruption of normal development: significant delays in larval moults, appearance of permanent larvae in the 3rd and 6th instars, and abnormal pupation. A possible interference with hormonal activity is discussed.     

Parasitization by Cotesia plutellae enhances detoxifying enzyme activity in Plutella xylostella

Insecticidal tests using diazinon showed that the mortality of Plutella xylostella larvae parasitized by Cotesia plutellae was reduced by 4.6-fold compared to that of the nonparasitized hosts. The use of chemicals with synergistic effect to insecticides in toxicity assay helps to elucidate the kind of enzyme involved in lowering insect mortality. Synergism of diethyl maleate and piperonyl butoxide with diazinon resulted to 2.4- and 1.9-fold increase, respectively, in susceptibility of parasitized larvae compared to those of nonparasitized larvae. These results indicated the possibility that the decrease in susceptibility to diazinon was due to the elevated activities of glutathione-S-transferase (GST) and cytochrome P450 monooxygenase (CYP), respectively. The GST activities in parasitized larvae were significantly higher than those of nonparasitized ones starting from three days post-parasitization until emergence of parasitoid larva. High GST activities during late parasitism could be attributed to both enzyme activities toward diazinon of parasitized P. xylostella larva itself and C. plutellae larva inside larval host. High GST activity one day after parasitization, although statistical significance was not detected, was caused by polydnavirus (PDV) and the venom of C. plutellae not by parasitoid larvae. Artificial injection of PDV plus venom demonstrated that the resulting increase in GST activity is similar to the increase brought by parasitization. High CYP activity after 3 days post-parasitization in parasitized larva was attributed mainly to the activity of parasitoid larva. Carboxylesterase activity in the parasitized host remained at a high level, while that in the nonparasitized host decreased slightly as pupation approaches. On the other hand, acetylcholinesterase activity also remained constant after parasitization until larval emergence, while that of the nonparasitized hosts decreased gradually as the host larvae approach pupation. These results were supported by inhibition tests using diazoxon in vitro.     

Biological and biochemical aspects of the disruption of adult formation in Tribolium by a novel bis(thiocarbamate) R-31026 and the juvenoid R-20458

S,S-Di-isobutyl N ethylethylenebis(thiocarbamate) (R-31026) had a strong morphogenetic effect, acting by contact on the newly formed pupae of Tribolium confusum and Tribolium castaneum and thereby disrupting adult development. At a dietary concentration of 20 mg kg^?1, 95% of pupae of T. confusum produced pupa-adult intermediates; T. castaneum pupae were affected to a smaller extent. Topical application with 0.002 μg per 0–24-h-old pupa of T. confusum resulted in the formation of 88% malformed intermediates. The larvae brought into contact with R-31026 were not affected and pupated normally, hence the compound differs in its activity from that of a typical juvenile hormone compound. On the other hand, the pupal morphogenetic activity of R-31026 resembles that of the typical juvenoid compound 3-[5-(4-ethylphenoxy)-3-methylpent-3-enyl]-2,2-dimethyloxirane (R-20458). The progeny of the emerging adults from pupae treated with either R-20458 or R-31026 were strongly affected. The effective dosages were far below those required for pupal morphogenetic activity. Biochemical studies showed an increase in the soluble protein fraction during the pupal stage after treatment with either R-31026 or R-20458 indicating disturbances in protein build-up. The bis(thiocarbamate) R-31026 has more favourable practical properties than R-20458 for controlling agricultural insects, because it does not prolong the larval feeding stage.     

Laboratory evaluation of the insect growth regulator lufenuron againstHelicoverpa armigera on cotton

An insect growth regulator (IGR), lufenuron (Match 5EC), was tested for its toxicity toHelicoverpa armigera on cotton. Potency of the IGR against the larval stage of the pest was demonstrated with respect to larval instars; the LC₉₀ values of 1st, 2nd, 3rd, 4th and 5th instar larvae were 5.63, 7.89, 8.03, 11.39 and 14.76 mg a.i.l ⁻¹, respectively. However, different larval instars did not differ significantly with respect to LC₅₀ and LC₁₀. IGR-treated larvae had swollen heads and were significantly smaller (1.5–2.3 mm) than the untreated control (2.9 mm). Larval weight was significantly reduced from 190 mg in the control to 50–70 mg in the lufenuron treatment. IGR treatment in the larval stage significantly affected both pupal length and pupal weight. Pupal duration of the test insect was significantly extended by IGR treatment. Pupal deformities, including an inability to shed the last larval skin and formation of larval-pupal intermediates, occurred following treatment. A significant reduction in adult emergence was recorded. In addition, abnormalities in the form of development of cavities in the forewings of adult were evident. A significant decline in fecundity was noted in the studies. http://www.phytoparasitica.org posting Feb. 3, 2003.